10.1515 - CCLM 2023 7047

Poster Abstracts – WorldLab • EuroMedLab Roma 2023 – Rome, Italy, May 21-25, 2023 • DOI 10.
1515/cclm-2023-7047
Clin Chem Lab Med 2023; 61, Special Suppl, pp S87 – S2222, May 2023 • Copyright © by Walter de Gruyter • Berlin • Boston S1015
Endocrinology
P0928
EVALUATION OF NEUTROPHIL TO LYMPHOCYTE RATIO (NLR) AND PLATELET TO LYMPHOCYTE RATIO (PLR) AS
INFLAMMATORY MARKERS IN PATIENTS WITH TYPE 2 DIABETES MELLITUS
S. Tyagi 1
1
Department of Biochemistry, SRVS Government Medical College, SHIVPURI (M.P.) INDIA
BACKGROUND-AIM
Diabetes Mellitus (DM) is a serious threat to global health with an increasing prevalence. DM causes serious
complications such as diabetic nephropathy, diabetic retinopathy, and diabetic neuropathy. Innate immunity and
inflammatory mediators play an important role and could lead to insulin resistance. The present study aimed to
evaluate the NLR and PLR in T2DM patients in comparison with healthy controls.
METHODS
This prospective study was conducted in SRVS Government Medical College, Shivpuri, Madhya Pradesh, India. In this
study, a total of 220 subjects were involved, among them 110 were T2DM patients as cases and 110 healthy subjects
as controls. This study was approved by the institutional ethics committee and informed consent was obtained from
all the study subjects. Demographic details, physical and clinical examination were done for all the study subjects.
Under aseptic conditions, 5 ml of fasting venous blood samples were collected from all subjects and aliquoted into
plain (3ml) and EDTA (2 ml) tubes and allowed to stand for 1 hr and centrifuged at 3000 rpm for 10 minutes at 4° C to
obtain the clear serum sample. The obtained serum sample was used for the estimation of fasting and post-prandial
glucose, renal profile, lipid profile using commercially available autoanalyzer kits. Whole blood sample was used for
the Complete Blood Count (CBC) in EDTA vials. NLR and PLR were calculated. Whole blood sample was used for the
estimation of HbA1c and Body Mass Index (BMI) was calculated.
RESULTS
In the current study, it is observed that in the study subjects NLR 6.7±2.3 and PLR 19.1±6.9 were significantly increased
in T2DM cases whereas HDLC 30.7±3.9 mg/dl was significantly decreased in T2DM cases.
CONCLUSIONS
The present study results may conclude that increased NLR and PLR may be used as a marker for inflammation in T2DM.
These may serve as an alternative to the other costly inflammatory markers.
Poster Abstracts – WorldLab • EuroMedLab Roma 2023 – Rome, Italy, May 21-25, 2023 • DOI 10.1515/cclm-2023-7047
Endocrinology
P0929
LEVEL OF AGREEMENT AND CORRELATION BETWEEN THE ESTIMATED HEMOGLOBIN A1C RESULTS DERIVED BY
CONTINUOUS OR CONVENTIONAL GLUCOSE MONITORING SYSTEMS COMPARED WITH THE POINT-OF-CARE OR
LABORATORY-BASED MEASUREMENTS
A. Al Hayek 1, S. Sobki 2, S. Sobki 2

1
Department of Endocrinology and Diabetes, Diabetes Treatment Center, Prince Sultan Military Medical City, Riyadh, Saudi
Arabia
2
Saudi Society for Clinical Chemistry (SSCC), PO BOX 22490. LABORATORY. Riyadh, Saudi Arabia
BACKGROUND-AIM
This study aimed to investigate the level of agreement between the standard laboratory test for HbA1c (Lab-HbA1c)
with point-of-care testing (POCT-HbA1c) and glucose monitoring index (GMI) derived by intermittently scanned CGM
(isCGM) or estimated average glucose (eAG) derived by conventional
self-monitored blood glucose (SMBG) devices.
METHODS
Between May and December 2020 with 81 patients with diabetes who used the isCGM system (n = 30) or conventional
finger-pricking SMBG system (n = 51). At the same visit, venous and capillary blood
samples were taken for routine HbA1c analysis by the standard laboratory and POCT methods, respectively. Also, for
isCGM users, the GMI data for 28 days (GMI-28) and 90 days (GMI-90) were obtained, while for SMBG users, eAG data
for 30 days (eAG30) and 90 days (eAG-90) were calculated. The limits of agreement in different HbA1c measurements
were evaluated using a Bland-Altman analysis.
RESULTS
Based on the Bland-Altman analysis, HbA1c levels for 96.7% and 96.1% of
the patients analyzed by the POCT and the standard laboratory methods were within
the range of the 95% limit of agreement in both isCGM and conventional SMBG users,
respectively. About 93.3% of the GMI measurements were within the 95% limit of
agreement. Also, about 94.12% of the eAG-30 and 90.2% of the eAG-90
measurements were within the 95% limit of agreement. Moreover, the correlation
analysis revealed a statistically significant positive correlation and linear regression
among Lab-HbA1c, POCT-HbA1c, GMI, and eAG in both conventional SMBG and
isCGM users (all p < 0.001). These positive results persisted significantly after
adjusting for different factors (all p < 0.001).
CONCLUSIONS
GMI derived by isCGM or eAG derived by conventional SMBG systems,
as well as the POCT-HbA1c measurements, showed a high level of agreement;
therefore, we recommend them as potential methods for diabetes monitoring, especially
when a rapid result is needed or with patients with uncontrolled diabetes or on
intensive insulin therapy.
Endocrinology
P0930
IMMUNOMODULATORY EFFECTS OF VITAMIN D SUPPLEMENTATION IN A DEFICIENT POPULATION
E. Sharif 1, S. Al Khodor 2
1
Department of Biomedical sciences , college of health and sciences, Qatar university
2
Research Department, Sidra Medicine, Doha
BACKGROUND-AIM
In addition to its canonical functions, vitamin D has been proposed to be an important mediator of the immune
system. Despite ample sunshine, vitamin D deficiency is prevalent (>80%) in the Middle East, resulting in a high rate of
supplementation. However, the underlying molecular mechanisms of the specific regimen prescribed and the potential
factors affecting an individual's response to vitamin D supplementation are not well characterized. Our objective is to
describe the changes in the blood transcriptome and explore the potential mechanisms associated with vitamin D3
supplementation in one hundred vitamin D-deficient women who were given a weekly oral dose (50,000 IU) of vitamin
D3 for three months.
METHODS
A high-throughput targeted PCR, composed of 264 genes representing the important blood transcriptomic fingerprints
of health and disease states, was performed on pre and post-supplementation blood samples to profile the molecular
response to vitamin D3. We identified 54 differentially expressed genes that were strongly modulated by vitamin D3
supplementation.
RESULTS
Network analyses showed significant changes in the immune-related pathways such as TLR4/CD14 and IFN receptors,
and catabolic processes related to NF-kB, which were subsequently confirmed by gene ontology enrichment analyses.
We proposed a model for vitamin D3 response based on the expression changes of molecules involved in the receptor-
mediated intra-cellular signaling pathways and the ensuing predicted effects on cytokine production. Overall, vitamin
D3 has a strong effect on the immune system, G-coupled protein receptor signaling, and the ubiquitin system.
CONCLUSIONS
We highlighted the major molecular changes and biological processes induced by vitamin D3, which will help to further
investigate the effectiveness of vitamin D3 supplementation among individuals in the Middle East as well as other
regions.
Endocrinology
P0931
THE IMPORTANCE OF SERUM ANDROGEN LEVELS IN EARLY DETECTION OF ADRENOCORTICAL CARCINOMA. A CASE
REPORT.
J. Guillén Reyes 1, S. Muñoz Madrid 1, Y. Posada Franco 1, C.M. Amor Llamas 1, M. Molina Fernández-Posse 1, M.D. Ibáñez
Royo 1, M.J. Torrejón Martínez 1, M. Martínez-Novillo González 1
1
Clinical Analysis Department, Laboratory Medicine Institute, Hospital Clínico San Carlos, Madrid, Spain
BACKGROUND-AIM
Adrenocortical carcinoma (ACC) is an extremely rare disease originated in the cortex of the adrenal gland that has an
incidence of 0.7-2 cases/million habitants/year, leading to 0.2% of all cancer deaths. It can occur at any age, being
more frequent in the first and fifth decade of life.
METHODS
We present the case of a 39-year-old woman with amenorrhea, hirsutism and acne of 8 months of evolution with no
other relevant clinical findings. A hormone analysis carried out 12 months before showed no abnormalities.
RESULTS
A blood test and a transvaginal ultrasound were requested for further examination. The transvaginal ultrasound did not
show any pathological findings, but the blood test showed abnormal androgen levels in serum: δ-Androstenedione >10
ng/mL [0.3-5.3], dehydroepiandrosterone sulfate (DHEA-S) = 11 µmol/L [0.48-10.6] and total testosterone = 13 ng/mL
[0.3-2.6]. Laboratory results were reported urgently. A CT scan showed a heterogeneous solid mass of 7.69x9.58x9.16
cm in its transverse, craniocaudal and anteroposterior axis, respectively, in the right adrenal gland.
After a laparoscopic adrenalectomy, the anatomic pathology examination reported a high-grade ACC with
lymphovascular invasion. The genetic study detected two somatic variants in the tumour: c.1444del (p.Glu482Lysfs*82)
in MEN1 and c.782+2_782+5delTCAG in TP53, as well as RB1 loss. She started chemotherapy with mitotane. A blood
test performed 2 months later showed undetectable androgen levels: δ-Androstenedione = 0.4 ng/mL [0.3-5.3], DHEA-
S = 0.05 µmol/L [0.48-10.6] and total testosterone <0.3 ng/mL [0.3-2.6].
Six months later the patient started again with hirsutism, amenorrhoea and arterial hypertension, presenting high
levels of androgens in blood. She developed liver and lung metastases and died a few weeks later.
CONCLUSIONS
ACC is an aggressive malignancy with poor prognosis and high risk of recurrence, with five-year overall survival being
below 40%. The measurement of androgen serum levels, along with the appropriate imaging tests, is useful in early
ACC diagnosis. Early detection and treatment are key to improve the prognosis and to help increase overall survival.
Monitoring androgen levels is also essential for the evaluation of therapy success and for the detection of cancer
recurrence.
Endocrinology
P0932
VARIATION IN LIPID BALANCE ACCORDING TO HOMOCYSTEINE LEVEL
S. Fendri 2, M. Boudaya 2, R. Ben Salah 1, K. Jamoussi 2, Z. Bahloul 1

1
CHU Hedi Chaker, internal medecine departement, sfax
2
CHU Hedi Chaker, laboratory of biochemistry, Sfax
BACKGROUND-AIM
Homocysteine is an intermediate sulfur amino acid in the metabolic pathways of cysteine production from methionine,
which is increasingly found to be involved in different pathological processes. We are interested in the course of our
work to study its implication on lipid balance in patients with venous thrombosis (DVT).
METHODS
This is a case-control observational study, comparing 47 healthy controls with 47 patients admitted to the internal
medicine department for the management of DVT. The assay of homocysteine was done by an enzymatic method. The
determination of total cholesterol and triglycerides was done according to an enzymo-colorimetric method. HDLc was
determined using an end-point enzyme solubilization method.
RESULTS
The mean age of patients and controls was 40.8 ± 10.5 years. Both groups included 27 men (57.5%) and 20 women
(42.5%).
Homocysteine was significantly (p<0.001) higher in ill subjects (mean 17.42 ± 5 µmol/L) compared to controls (mean
9.41 ± 3.1 µmol/L). Statistical analysis showed that homocysteine was positively correlated with LDLc (p=0.002; r=0.392)
and negatively with HDLc (p=0.002; r=-0.392).
CONCLUSIONS
Homocysteine is a sulfur amino acid involved in many pathological processes, this can be partly explained by the effect
on lipid metabolism with a decrease in HDLc and an increase in LDLc with their cellular toxic effects.
Endocrinology
P0933
MULTIPLE ENDOCRINE NEOPLASIA TYPE 1 (MEN1): CASE REPORT.
B. Nafría Jiménez 1, L. Alcázar García 2, M. Linares Torres 2, S. Ouriach Makrini 2, S. Jiménez Sánchez 2, L. Caja Guayerbas
2
, A. Cortés Gómez 2, E. Sancho Fitos 2, L. Martínez Cid 2
1
Clinical Biochemistry Department, Donostia University Hospital, San Sebastian, Spain
2
Clinical Biochemistry Department, Ramón y Cajal University Hospital, Madrid, Spain.
BACKGROUND-AIM
Multiple Endocrine Neoplasia (MEN) are autosomal dominant inherited conditions, characterised by a predisposition
to the appearance of various neuroendocrine tumours. 3 types of MEN syndromes are most common:
1. MEN 1: mutations that inactivate MEN1 tumour suppressor gene. Parathyroid, anterior pituitary gland and
gastroenteropancreatic neuroendocrine tumours.
2. MEN2: mutations that activate RET protooncogene.
a.MEN2a: medullary thyroid carcinoma (MTC), pheochromocytoma, and primary hyperparathyroidism (PP).
b.MEN2b: MTC and pheochromocytoma. Rarely PP.
3. Familial form of medullary thyroid cancer.
METHODS
35-year-old female came to emergency department due to sweating and onset of hypertensive crisis. Personal records:
pancreatic nodule on surveillance.
Laboratory analysis: hypercalcemia and increased circulating parathormone (PTH) concentration (105 pg/mL;
VR[10-65]). Computed axial tomography: 2 lower parathyroid adenomas, confirming PP, and a total parathyroidectomy
was performed. No pituitary adenomas.
RESULTS
Repeated laboratory analysis, 6 months later: elevated gastrin (120 pg/mL; VR[13-115]), chromogranin A (726 ng/mL;
VR[0-102]) and glucagon (305 pg/mL; VR[0-209]) levels. Hypo-PTH post-surgical.
Suspecting MEN1, a Nuclear Magnetic Resonance was performed, visualizing 2 peripancreatic lymphadenopathies.
Treatment: biliopancreatic surgery.
Molecular study of the MEN1 gene (by Sanger sequencing): identification of the pathogenic mutation
p.F369_V372delinsL (chromosome 11q13), genetically confirming the diagnosis of MEN1. Genetic testing was done
to first-degree family members, finding the same mutation in heterozygosis in her 2-year-old daughter (no clinical
manifestations yet).
CONCLUSIONS
A timely and accurate diagnosis of MEN1 is paramount to improve disease outcomes. For this purpose, an early
identification of the tumour manifestations accompanied by the analysis in the Clinical Laboratory of the hormonal
level and the genetic study are crucial.
Genetic screening allows the detection of affected relatives before they present symptoms and thus early treatment
can be administered. In this case, and given the daughter´s involvement, genetic counselling was given to the couple
to perform preimplantation genetic diagnosis (PGD) when they decide to increase their offspring.
Endocrinology
P0934
CHANGES OF TSH, FT3 AND FT4 LEVELS IN PATIENTS WITH HYPERTHYROIDISM
N. Serdarevic 1, A. Redzepovac 1
1
Institute for Clinical Biochemistry and Immunology University of Sarajevo Clinics Center, Sarajevo, Faculty of Pharmacy,
University of Tuzla, Bosnia and Herzegovina, Faculty of Health Sciences, University of Sarajevo, Bosnia and Herzegovina
BACKGROUND-AIM
The aim of our study was to determine the serum concentration of TSH, FT3, and FT4 in patients with latent and
expressed hyperthyroidism and compare the results to determine the differences between the mild hyperthyroidism
and hyperthyroidism
METHODS
The study included 1200 patients age (30-40) years, divided into three groups that are into a control group with healthy
patients, groups with respondents who have hyperthyroidism and a group of patients with mild hyperthyroidism. The
concentration of the hormone TSH, FT3 and FT4 were analyzed using IMMULITE 1 Siemens.
RESULTS
The patients with hyperthyroidism had an average concentration of TSH 0.016 mIU/L, FT3 11.9 pg/mL, FT4 51.3 pmol/
L. The group with mild hyperthyroidism had an average concentration of TSH 0.08 mIU/L, FT3 3.773 pg/mL, FT4 17.2
pmol/L. Man Whitney's U test showed that the value of parameter TSH between our group investigated the existence
of significant statistical differences as between the control group and the group with hyperthyroidism (Z = 6.745;
p <0.001) and between the control group and the group with mild hyperthyroidism (Z = 6.655; p <0.001). For FT4,
we completed a statistically significant difference between the control group and the group with hyperthyroidism
(Z = 6.625; p <0.005) and between groups with hyperthyroidism and mild hyperthyroidism (Z = 1.747; p <0.002).
In our study the significant difference between concentrations FT3 between the control group and the group with
hyperthyroidism (Z = 6.65; p <0.001) and between groups with hyperthyroidism and mild hyperthyroidism (Z = 2.832 ;
p <0.005). Sensitivity TSH hyperthyroidism group and the control group was 98.5 % with the specificity of 90.5 %, while
the sensitivity was 98.3 FT3 with a specificity of 86.7 %, sensitivity was 99.3 % FT4 with a specificity of 92.3 %. AUC for
TSH was 0.935, while for FT3 it totaled 0.847 and for FT4 was 0.856.
CONCLUSIONS
In patients with latent and expressed hyperthyroidism serum hormone TSH decreased while the concentration of FT3
and FT4 elevated in comparison to the control group. The concentration of TSH is reduced in both hyperthyroidism
and mild hyperthyroidism. Serum concentrations of FT3 and FT4 were elevated in hyperthyroidism, while in mild
hyperthyroidism serum concentration of FT3 and FT4 were in the reference area.
Endocrinology
P0935
PREVALENCE OF THYROID DISEASE IN PREGNANT WOMEN IN OUR HEALTH AREA.
M. Zárate Bertolini 1, M.D.C. Esteban De Celis 1, M. Giménez Blanco 1, R. Rubio Sánchez 1, J. Bobillo Lobato 1
1
Hospital Universitario Virgen de Valme, Sevilla
BACKGROUND-AIM
Thyroid disorders during the first trimester of pregnancy often go unnoticed due to the non-specific symptoms of
pregnancy. Among the main functions of the thyroid gland during pregnancy is the correct development of the brain
and the central nervous system of the fetus. During the first weeks of gestation, the fetus depends exclusively on the
mother to produce thyroid hormones, so untreated or inadequately treated thyroid disease during early pregnancy has
been associated with adverse obstetric and fetal outcomes, such as poor brain development, premature birth, low birth
weight, preeclampsia, placental anomalies, postpartum hemorrhage.
Due to the importance and prevalence of this pathology, universal screening of pregnant women with specific reference
values for the first trimester of pregnancy is used in our laboratory.
The aim was to identify the prevalence of thyroid disease in pregnant women during the first trimester of pregnancy
in our health area.
METHODS
Retrospective study in which the results of the thyroid dysfunction screening during the first trimester of pregnancy
in 2021 were extracted from the SIL of the laboratory. A total of 2533 pregnant women were obtained, who underwent
serum thyrotropin levels (TSH), free thyroxine (FT4). Hospitalized patients and urgent requests were excluded. The
reference values used were: TSH: 0.17 – 4.40 µIU/mL, FT4: 0.80- 1.80 ng/dL The samples were processed in the Roche®
Cobas e801 using an electrochemiluminescence immunoassay and statistical data analysis using MedCalc v .13.3.0.0.
RESULTS
2533 pregnant women were studied, of which 262 presented thyroid pathology with a prevalence of 10.34%, with
the following distribution: The prevalence of subclinical and overt hypothyroidism was 6.04% and 0.20% respectively,
while subclinical and overt hyperthyroidism was 3.55% and 0.55% respectively. Patients with manifest hypothyroidism
and hyperthyroidism were more common in patients between 26 and 30 years of age, while pregnant women with
hypothyroidism and subclinical hyperthyroidism in the 31 to 35-year-old group.
CONCLUSIONS
The prevalence of thyroid pathology in pregnant women in our health area is 10.34%, which is like other studies carried
out. The importance of early diagnosis of thyroid pathology predominates despite not being such a frequent pathology.
Endocrinology
P0936
COPEPTIN AND ITS ROLE IN THE DIAGNOSIS OF DIABETES INSIPIDUS.
B. Nafría Jiménez 1, A. Garrido Chércoles 1, M.J. Izquierdo Vicente 1, R. Cabezón Vicente 1, E. Bereciartua Urbieta 1, P.
Pascual Usandizaga 1, C. Lallave Hernández 1, N. López Barba 1
1
Clinical Biochemistry Department, Donostia University Hospital, San Sebastian, Spain.
BACKGROUND-AIM
Diabetes insipidus (DI) is a disorder of water metabolism resulting from a deficient action of antidiuretic hormone
(AVP). DI must be further differentiated into central (DIC), if the alteration is the AVP-synthesis in the hypothalamus-
neurohypophysis; or nephrogenic (DIN), due to renal insensitivity. Also, it must be differentiated with diabetes mellitus
and primary polydipsia (PP).
Occasionally, it is necessary to perform the water deprivation test, but its interpretation can be uncertain and poorly
tolerated. However, the basal measurement of copeptin (C-terminal of AVP precursor) is very useful in the diagnosis of
DI, since it is synthesized in equimolar quantity with AVP, has a high ex vivo stability and an automatable measurement.
METHODS
Patient 1.55-year-old female, polydipsia and polyuria for 2 months. Blood analysis:Osm 326 mOsm/kg [VR 275-301], Na
+ 148 mEq/L [135-145], K+ 5.1 mEq/L [3.3-5.1]. Urine:Osm 381 mOsm/kg [50-1200], Na+ 53 mmol/L [80-180], diuresis
3300 mL/24h [500-2500].
Patient 2.42-year-old male, polydipsia and polyuria for 6 months. Blood analysis:Osm 430, Na+ 146, K+ 4.5. Urine:Osm
430, Na+ 75, diuresis 4950.
Patient 3.47-year-old female, polydipsia and polyuria for 1 year. Blood analysis:Osm 278, Na+ 142, K+ 5.2. Urine:Osm
309, Na+ 50, diuresis 4550.
RESULTS
Plasma copeptin assay: immunoluminescence, VR [1-14 pmol/L]
Patient 1.Copeptin: basal 2.8; after hypertonic saline infusion 2.9. Diagnostic: DIC. Treatment:desmopressin
Patient 2.Copeptin: basal 25. Diagnostic: DIN. Treatment:diuretics.
Patient 3.Copeptin: basal 2.6; after hypertonic saline infusion 78.5. Diagnostic: PP. Treatment:hydric restoration.
CONCLUSIONS
DI is a rare but treatable condition that typically presents polydipsia and polyuria, but these symptoms do not
differentiate it from other water disorders.
Copeptin constitutes a stable and reliable surrogate marker for AVP, whose levels correlate strongly with plasma
osmolality. Furthermore, it can be measured in clinical routine with available assays with high-level technical
performance. In this way, several limitations with respect to other classic tests are overcome and, as occurs in these
patients, copeptin-based diagnostic algorithm is proposed to reliably differentiate between the distinct entities.
Endocrinology
P0937
VITAMIN D OF SERUM LEVELS EVALUATION STUDY IN JORDANIAN POPULATION
H. Harirah 1
1
Department of Medical Laboratory Sciences, Faculty of Allied Medical Sciences, Zarqa university, Zarqa
BACKGROUND-AIM
This study aimed to investigate serum 25(OH)D levels in a sample of the Jordanian population according to age and
gender in Amman, Jordan.
METHODS
Cross-sectional retrospective study was performed to determine the prevalence of vitamin D 25-hydroxy vitamin D (25-
OHD) levels among 4133 apparently healthy Jordanian people.
RESULTS
The study results on Vitamin D levels showed that the prevalence of the 0–2-year group was 1%. The mean ± SD of
vitamin D in the 0-2 years group was 47.5 ± 24.5, which is considered to be within the normal range, while the spread
of the 3-17 age group was 7.5% of the mean ± SD of vitamin D in the 3-17 years group was 19.2 ± 24.7 which considered
being a moderate deficiency, on the other hand, the percentage of >18 group was 91.5 % the mean ± SD of vitamin D
in this group was 22.9 ± 14.0 which is considered to be a mild deficiency.
CONCLUSIONS
There was a high prevalence of vitamin d deficiency, and more efforts should be made to encourage the population
to be awarded beneficial nutrition and tappets, also more studies are recommended to reevaluate the normal ranges
and genetic deposition in Jordan.
Endocrinology
P0938
OPTIMIZATION OF THE REFERENCE INTERVALS FOR SERUM TOTAL VITAMIN B12
J.A. Delgado RodrÍguez 1, M.I. Pastor GarcÍa 1, N. MÁrquez JimÉnez 1, G. Costa Petit 1, A. RegÍs PerellÓ 1
1
Department of Laboratory Medicine, Hospital Universitari Son Espases, Palma de Mallorca, Spain
BACKGROUND-AIM
The establishment of appropriate reference intervals for the assisted population is crucial for the correct interpretation
of laboratory results. In this line, indirect or pseudodirect methods can be used as an alternative to traditional methods.
As for vitamin B12, its deficiency is frequent in all age groups and is closely related to a series of chronic diseases.
Our aim was to obtain the reference intervals of vitamin B12 for adult population (>15 years), using pseudodirect
methods.
METHODS
This is a prospective study. All Primary Care laboratory requests from individuals >15 years were considered if they
included vitamin B12, and the results for complete blood count, creatinine, ALT, total bilirubin and folate were within
their respective reference intervals, as well as urine protein (dipstick) negative and eGFR (CDK EPI2009) >90ml/
min/1.73m2. Besides, medical records were reviewed and those individuals who presented supplementation with
vitamin B12 or folate, any pathology or treatment that could cause cobalamin metabolism disorders, and pregnant
women were excluded.
On the other hand, those individuals whose vitamin B12 result presented antibody-mediated interference in the Alinity
i immunoassay (Abbot Diagnostics) (vitamin B12 after PEG precipitation <70%) were excluded.
Finally, urinary methylmalonic acid (MMA) determination [mmoL MMA / moL creatinine (urine)] was extended to all
individuals who met the above criteria. Individuals whose results were within the reference intervals for this magnitude
were included.
A normality test (Kolmogorov-Smirnov) was performed for the vitamin B12 variable. Given the non-normality of the
distribution, a Box-Cox transformation (MEDCALC software) was performed. Requests were grouped by age (15–29; 30–
49; 50–69; >69) with a total of 120 individuals per subgroup. Differences between partitions were evaluated using the
Harris-Boyd methodology. Reference intervals were determined by parametric calculation.
RESULTS
The reference interval for vitamin B12 does not depend on age. We consider the following reference limits for our adult
population: Lower Limit 146pmol/L; Upper Limit 554pmol/L.
CONCLUSIONS
The adequacy of the vitamin B12 reference intervals would lead to a reduction in the number of additional tests and
would avoid incorrect medical actions.
Endocrinology
P0939
RELATIONSHIP BETWEEN ERECTILE DYSFUNCTION AND TYPE 2 DIABETES
B. Nouala 1, L. Mbouche 3, M.C. Etoa 5, J. Yondo 1, J.R. Nkeck 4, F. Amazia 1, C. Mbakop 1, A. Ntyam Abena 1, A. Batakeh
B 6, V.J. Ama Moor 2
1
Department of Biochemistry – Faculty of Medicine and Biomedical Sciences -University of Yaounde 1, Cameroon
2
Department of Biochemistry – Faculty of Medicine and Biomedical Sciences -University of Yaounde 1, Cameroon ;
Laboratory of Biochemistry – Yaounde University Teaching Hospital, Cameroon
3
Départment of Chirugical Sciences, Urology Unit - Faculty of Medicine and Biomedical Sciences -University of Yaounde 1,
Cameroon
4
Départment of Internal Medicine - Faculty of Medicine and Biomedical Sciences -University of Yaounde 1, Cameroon
5
Départment of Internal Medicine, Endocrinology Unit - Faculty of Medicine and Biomedical Sciences -University of Yaounde
1, Cameroon
6
Woodhull medical centre/Department of Medicine, Brooklyn, New York, USA
BACKGROUND-AIM
Erectile dysfunction (ED), the most common sexual disorder in type 2 diabetes mellitus (T2DM), has been shown to be
caused by endothelial. Homocysteine (Hcy) is one of the early biomarkers of endothelial damage. This study sought to
evaluate the relationship between homocysteine and erectile dysfunction in men with type 2 diabetes at the Yaounde
Central Hospital.
METHODS
We conducted a cross-sectional analytic study in a population of consenting adult men with T2DM; over a period of
7 months from January to August 2022. Subjects with conditions capable of inducing ED or influencing Hcy were
excluded. The IIEF-5 score was used to assess the frequency/severity of ED. Hcy assay on plasma was done by
competitive ELISA. Factors associated with ED, and the relationship between Hcy and ED were investigated by bi- and
multivariate analyses followed by logistic regression. Significance was considered at p<0.05.
RESULTS
We included 62 T2DM patients. The mean age was 50.5±9.8 years. Fifty percent (31/62) had erectile dysfunction, with 15
(48.3%) being severe. ED was significantly associated with diabetes duration >2years [OR = 17.3(4.9-61.2), p<0.001] and
poor glycemic control (HbA1C >8%) [OR = 4.4(1.5-12 .8) p=0.005]. Although hyperhomocysteinemia was found in 88.7%
(n=55) of the participants, there was a significant difference in homocysteine levels between patients with and without
ED (p=0.04). There was a strong association between hyperhomocysteinemia and ED [OR=0.53 (0.44-0.97); p=0.04].
CONCLUSIONS
Erectile dysfunction affects nearly half of men with diabetes. The association between homocysteine and ED would
suggest endothelial damage in our population.
Key words: type 2 diabetes mellitus; erectile dysfunction; homocysteine
Endocrinology
P0940
PSEUDOHYPOPARATHYROIDISM TYPE IB: CLINICAL-BIOCHEMICAL DIAGNOSIS AND MOLECULAR CONFIRMATION
B. Nafría Jiménez 1, A. Garrido Chércoles 1, M.J. Izquierdo Vicente 1, R. Cabezón Vicente 1, B. Álvarez Iturregui 1, A.
Ulazia Garmendia 1, M. Echeverria Urroz 1, D. Álvarez Nistal 1
1
BACKGROUND-AIM
Pseudohypoparathyroidism (PHP) is a rare inherited disorder characterized by hypocalcemia, hyperphosphatemia and
hyperparathyroidism secondary to organ resistance to parathyroid hormone (PTH).
This entity is usually diagnosed in childhood, especially due to high calcium levels or to presenting a typical phenotype
known as Albright's hereditary osteodystrophy (AHO). Genetic and epigenetic alterations at the GNAS locus (20q13) are
responsible for the Gs-adenylate-cyclase protein dysfunctions. This region is subject to the imprinting phenomenon
so that, depending on the type and origin of the alteration, the following classification has been established: Type Ia,
Ic and II– mutation inheritante maternal; Type Ib– inheritante paternal.
METHODS
35-year-old female came to emergency department due to dizziness and muscle weakness. No previous pathologies.
Physical examination: low blood pressure. No AHO phenotype.
Laboratory analysis: corrected calcium 6.2 mg/dL [VR 8.6-10.2], phosphate 5.1 mg/dL [2.5-4.5], PTH 425 pg/mL
[10-65], TSH 11.2 mU/L [0.3-4.2], free-T4 0.82 [0.85-1.85]. Other parameters (renal function, magnesium, antithyroid
antibodies) were normal.
RESULTS
•Urine: hypocalciuria 49 mg/24 h [100-320].
•Hormonal study was completed: low estradiol <5 pg/mL, with normal cortisol concentration (30.5 µg/dL – excludes
adrenal deficiency or Addison's syndrome).
•Hand X-ray, brain CT and thyroid ultrasound were normal.
Given these results, PHPIb is suspected. The genetic study of the GNAS gene (MLPA) is extended:
•Patient: GNAS gene mutation, consistent with PHPIb.
•Parents: Father: presented the same mutation, in heterozygosis (paternal inheritance). Given the absence of AHO or
biochemical alteration, he had not been studied or diagnosed before. Mother: no mutation.
•Husband and daughter: no mutation.
Evolution: Patient was treated with Calcitriol and levothyroxine (Eutirox®), with disappearance of symptoms and
normalization of calcium levels at 8 months.
CONCLUSIONS
Although PHP is a rare endocrine disease, our case was detected in adulthood by laboratory results (hypocalcaemia
and hyperparathyroidism), with no-AHO phenotype. Likewise, its association with other hormonal alterations and
molecular studies allowed knowing its inheritance and confirming the diagnosis.
Endocrinology
P0941
WHEN TECHNOLOGY MAKES THE DIFFERENCE: PREVALENCE OF MALE HYPOGONADISM AMONG HIV PATIENTS
CHANGES WHETHER TESTOSTERONE IS MEASURED BY MASS-SPECTROMETRY (LC-MS/MS) OR BY IMMUNOASSAY (IA)
L. Roli 1, S. Tagliavini 2, M.C. De Santis 2, S. De Vincentis 3

1
Department of Laboratory Medicine and Pathology, Azienda USL Modena, Italy
2
Department of Laboratory Medicine and Pathology, Azienda USL of Modena, Modena, Ital
3
Unit of Endocrinology, Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia,
Modena, Italy
BACKGROUND-AIM
Multifactorial causes underlie the higher prevalence of hypogonadism in HIV-infected males; according to different
studies, it ranges from 13% to 40% in subjects aged from 20 to 60 years. HIV patients show normal levels of total
testosterone (TT), higher levels of sexs hormone binding globulin (SHBG) and lower levels of free testosterone (FT)
which is responsible for the biological activity of testosterone (T). The Endocrine Society guideline recommends
measurement of SHBG together with T and/or calculated FT (cFT) for the diagnosys of biochemical hypogonadism
(cutoff T > 320 ng/ml; cutoff cFT > 64 pg/ml).
METHODS
Serum samples were collected after an overnight fasting from 316 HIV-infected males aged 18 - 50 years, ongoing
antiretroviral therapy and not taking any androgen substitutive therapy. Serum TT was measured both with a
validated LC-MS-MS method and a chemiluminescent-immunoassay (CLIA) by Abbott, USA (Architect 2nd Generation
Testosterone); SHBG was assayed by Abbott Architect CLIA; cFT was calculated applying Vermeulen's equation to both
LC-MS/MS and CLIA results.
RESULTS
TT and cFT serum levels measured by LC-MS/MS were significantly lower than those obtained by CLIA (p< 0,0001); 6
(1,9%) patients had serum CLIA TT > 320 ng/ml but they resulted hypogonadal when serum TT was assessed by LC-
MS/MS; 9 (2.8%) patients showed CLIA cFT > 64 pg/ml but they were hypogonadal when cFT was calculated using LC-
MS/MS values.
Eugonadal HIV-infected patients showed significanly lower levels of serum SHBG than those with secondary or
compensated hypogonadism (p=0.001 and p=0.002 respectively); Prevalence of biochemical hypogonadism detected
by cFT was higher compared to serum TT (p<0.0001), regardless of the hormonal assay used.
CONCLUSIONS
The prevalence of hypogonadism in HIV-infected males assessed only on laboratory data was significantly higher when
serum TT was detected by LC-MS/MS than by CLIA, both considering serum TT (5.1% vs 3.2%, p<0.0001) and cFT (9.5%
vs 7%, p<0.0001) demonstrating how technologies can deeply impact on clinical assessment and consequently affect
decisions about therapy.
Endocrinology
P0942
FEMALE TESTOSTERONE IN IRELAND 2018-2022: THE EFFECTS OF SARS-COV-2.
K. Hutchinson 2, H. Ali 2, M. Louw 2, J. Faul 1

1
Connolly Hospital Blanchardstown, Dublin
2
Eurofins Biomnis, Dublin
BACKGROUND-AIM
Testosterone is an important hormone which in females is responsible for maintaining normal sexual and metabolic
functions. It also influences muscle and bone strength, urogenital health, mood, and cognitive abilities. In males, low
testosterone levels have been associated with severe SARS-CoV-2 illness, respiratory failure and death. In females,
studies show conflicting results. Several studies suggest that females are more likely to suffer from long COVID-19.
Female sex and 40–50 years of age are considered risk factors for long COVID. We need to understand the cause of sex
disparities and the role of androgens in women during pandemics.
METHODS
We have performed an audit of serum total testosterone measures in the healthy Irish female population during
2018-2022, including COVID pandemic (n = 19325). Total testosterone was measured in blood samples using the
radioimmunoassay (RIA) method. The software used for the statistical analysis was GraphPad Prism 5, Version 9.5.0.
Mean total testosterone (tT) concentrations were analysed overall before, during and after the COVID pandemic (and
specifically in relation to age groups: 18-39, 40-50 and >50).
RESULTS
The data showed statistically significant differences in testosterone concentrations in the age groups 18-39 and 40-50,
before, during and after the COVID-19 pandemic, but not in females over 50. The averages of tT were significantly low
in both groups in 2020 and 2021: for 18-39 group = 1.89, for 40-50 = 1.50 nmol/L; in 2022: for 18-39 group = 2.19, for
40-50 = 1.74 nmol/L. For post-menopausal women the averages of tT were 1.30 nmol/L in 2018-2019; and 1.32 nmol/
L in 2020-2021 and in 2022.
CONCLUSIONS
The decrease of tT in females <50 years old during the pandemic may be due to increased stress and lifestyle changes
in Ireland because of lockdowns. Increases in serum tT after the pandemic in pre-menopausal women might partly
explain the increased risk for long COVID as a mirror of higher inflammatory phenotype in this group. Further studies
are necessary to elucidate the importance of testosterone measures in females to enable us to fully examine changes
in tT status in Ireland during and after pandemics.
Endocrinology
P0943
CORTICOADRENAL CARCINOMA IN ELEVENTH-MONTH-OLD INFANT
J. Montenegro-Martínez 1, R. Sánchez-Rubio 1, E. Lepe-Balsalobre 1, J.M. Guerrero Montávez 1

1
Hospital Universitario Virgen del Rocío
BACKGROUND-AIM
Adrenocortical tumors are extremely rare in children and adolescents. According to estimates by the US National Cancer
Institute, only 0.2% of childhood processes are adrenocortical tumors. At ages as early as the one presented in this
case, figures are even lower. Therefore, the accumulated experience in diagnosis and follow-up is currently very limited.
METHODS
An 11-month-old female infant was referred to our hospital for a suspected cortisol-producing adrenal tumor. Physical
examination: BMI=28.98% (p>99), very high blood pressure values (up to 190/120 mmHg), cushingoid phenotype and
oropharyngeal and genital candidiasis. On palpation a lesion without defined borders is intuited at the level of the
left flank.
The following tests are requested: 1) Hormonal blood study: corticotropin=2.8 pg/mL, cortisol=24.4
µg/dL, aldosterone=1180 pg/mL, 17-hydroxyprogesterone=8.54 ng/mL, androstenedione= 12.30 ng/mL,
dehydroepiandrosterone sulfate=821.0 µg/dL, progesterone=5.74 ng/mL, testosterone=9.48 nmol/L; 2)
Catecholamines in urine: 5-hydroxyindoleacetic acid=7 mg/g, vanillylmandelic acid=183 mg/g, homovalinic acid=24
mg/g; 3) Neuron-specific enolase=63.1 ng/mL; 4) PET-CT: a metabolically positive left adrenal mass is detected.
Through supraumbilical transverse laparotomy, a bilobed tumor that encompasses the adrenal gland is resected. The
histopathological report describes an adrenal neoplasm, made up of large cells with ample cytoplasm, organized
in cords and nests surrounded by a fine vascular network. Adrenocortical neoplasia of intermediate category for
malignancy is diagnosed.
RESULTS
On day 12 post-surgical, a new hormonal analysis shows a decrease in its levels: cortisol=1.8 µg/dL, aldosterone=6.7
ng/dL, 17-hydroxyprogesterone=0.14 ng/mL, androstenedione<0.02 ng/mL, dehydroepiandrosterone sulfate=1µg/
dL, testosterone=0.09 ng/dL, progesterone=0.17 ng/mL. These results indicate adrenal insufficiency secondary to
adrenalectomy and confirm the need for exogenous corticosteroids.
CONCLUSIONS
Although clinical evidence and imaging tests are sufficient to establish the diagnosis and therapy, the determination
of the secretion profile of the tumor informs its degree of differentiation and it is of great value in post-surgical follow-
up and in the definition of remission/ recurrence.
Endocrinology
P0944
INCREASED ALL-CAUSE MORTALITY IN INDIVIDUALS WITH FUNCTIONAL VITAMIN D DEFICIENCY
M. Herrmann 1, S. Zelzer 1, M. Kleber 3, C. Drexler-Helmberg 2, P. Schlenke 2, P. Curcic 1, D. Enko 1, H. Scharnagl 1, S. Pilz

4
, W. März 5
1
Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Graz, Austria
2
Department for Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, Austria
3
Department of Internal Medicine 5 (Nephrology, Hypertensiology, Endocrinology, Diabetology, Rheumatology), Mannheim
Medical Faculty, University of Heidelberg, Mannheim, Germany
4
Division of Endocrinology and Diabetology, Department of Internal Medicine, Medical University of Graz, Graz, Austria
5
Synlab Academy, Synlab Holding Germany GmbH, Mannheim, Germany
BACKGROUND-AIM
25-hydroxyvitamin D [25(OH)D], the inactive prohormone of 1,25-dihydroxyvitamin D, represents the bodies vitamin
D reservoirs and is a predictor of mortality. The simultaneous determination of 25(OH)D, 24,25-dihydroxyvitamin D
(24,25[OH]2D) and calculation of the vitamin D metabolite ratio (VMR) allow a functional assessment of the vitamin D
status that may be superior to the use of 25(OH)D alone. This study investigated the association of functional vitamin
D deficiency and low 25(OH)D concentrations with all-cause mortality in the Ludwigshafen Risk and Cardiovascular
Health (LURIC) study.
METHODS
In 2456 baseline plasma samples from LURIC, 25(OH)D3, 25(OH)D2 and 24,25(OH)2D3 were measured by a validated
in-house LC-MS/MS method. The ratio between 24,25(OH)2D3 and 25(OH)D was calculated. 25(OH)D concentrations
<50 nmol/L were considered low. The criteria for functional vitamin D deficiency were a 24,25(OH)2D3 <3 nmol/L and a
VMR <4 % (the rounded upper limit of the first quartiles). Participants were followed over a median observation period
of 9.9 years.
RESULTS
In total, 653 (23.6 %) participants of LURIC died during follow-up. Participants with sufficient 25(OH)D concentrations
had an all-cause mortality of 16.9 % (136/803), whereas the absence of functional vitamin D deficiency was associated
with a mortality of 18.6 % (190/1024). Low 25(OH)D concentrations were associated with a mortality of 31.4 %
(519/1653). With 38.7 % (192/496), all-cause mortality was highest in individuals with functional vitamin D deficiency.
CONCLUSIONS
Only a subgroup of individuals with a low serum 25(OH)D concentration meets the criteria for functional vitamin
deficiency. These individuals have a higher risk of mortality than those with 25(OH)D concentrations of less than 50
nmol/L.
Endocrinology
P0945
ESTABLISHING REFERENCE INTERVALS FOR STEROID PROFILES IN BLOOD USING LIQUID CHROMATOGRAPHY-TANDEM
MASS SPECTROMETRY IN KOREAN ADULTS
H. Park 1
1
Samsung Medical Center
BACKGROUND-AIM
Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the most accurate and precise method for the
determination of testosterone concentrations. LC-MS/MS-based simultaneous measurement of endogenous steroid
hormones, which is less affected by cross-reactivity, could have advantages over the conventional immunoassay.
However, there has been no LC-MS/MS study concerning the reference interval and distribution of sex steroid hormones
including testosterone in the Korean population.
METHODS
A total of 266 Korean adults aged from 15 to 49 years were recruited including 137 females and 129 males. We
established reference intervals of the eight steroid hormones cortisol, cortisone, corticosterone, 11-deoxycortisol
(11DOC), androstenedione, 17OHPG, progesterone, and testosterone using an Infinity HPLC system coupled to a 1260
Agilent 6490 Triple Quadrupole Mass Spectrometer.
RESULTS
The reference intervals of androstenedione, testosterone, 17OHPG, and progesterone were separately established for
each sex. Notably, the reference interval of testosterone for Korean males was established as 7.35 nmol/L (90% CI,
5.74 nmol/L to 9.27 nmol/L) to 43.06 nmol/L (90% CI, 33.57 nmol/L to 50.74 nmol/L), after removing 5.4% (7/129)
of outliers. Our data showed that 12.4% of subjects were below the cutoff of low testosterone, and a prevalence of
approximately 2% with low testosterone less than 1.0 nmol/L was observed in Korean males aged between 15 and
49 years. Except for testosterone in males, other steroid hormones showed comparable distributions in males and
females, similar to previous studies for reference intervals of steroid hormones in the Caucasian population.
CONCLUSIONS
Given the prevalence of low testosterone, which is generally between 10% and 25% in elderly males, our study showed
10.40 nmol/L as the cutoff value of low testosterone in Korean adult males.
Endocrinology
P0946
REFERENCE INTERVALS FOR FREE T4 AND FREE T3 FOR ADULTS WERE ESTABLISHED USING THE INDIRECT SAMPLING
TECHNIQUE IN TWO LABORATORY CHAINS IN POLAND
E. Dworniczak 4, E. Zapala 4, J. Brąszkiewicz 5, M. Szczur 5, M. Nowacka 3, C. Lafrence 1, A. Ruzhanskaya 2

1
Beckman Coulter Inc, Chaska, USA
2
Beckman Coulter LLC, Moscow, Russia
3
Beckman Coulter LLC, Warsaw, Poland
4
DILAB Medical Diagnostic Nisko, Poland
5
VITALABO Medical Diagnostic Laboratories, Bydgoszcz, Poland
BACKGROUND-AIM
There has been a growing demand for the derivation of population-specific reference intervals (RIs) of commonly
ordered laboratory tests, especially related to the assessment of thyroid function. The direct method of RIs’
establishment provided in CLSI EP-28A3C is quite difficult and expensive. We have derived RIs for FT3 and FT4 for adults
in two big laboratory chains in Poland (Dilab, Lab1 and Vitalabo, Lab2) using the indirect technique and compared
them to those derived in Russia and provided in Beckman Coulter’s instruction for use (IFU).
METHODS
Based on publications, inclusion criteria of the retrospective study were defined as the presence of > 1.000 results of
FT4 and/or FT3, obtained from non-repeatable patients >18 yo of non-endocrinological departments with TSH within
the respective RI used in the laboratory. Around 165.000 results were tested for TSH, Free T3, and/or Free T4 total and
were determined to have normal TSH values as defined by a TSH reference interval of 0.38 – 5.33 µIU/mL. 95%, 97.5%,
and 99% RIs were calculated for Access Free T4 and Access Free T3 (Beckman Coulter, Inc.). For calculation, the JMP
software was used
RESULTS
RIs for FT3 and FT4 had no obvious differences between the two Polish labs. For FT4 (pmol/L, 95%), newly calculated
FT4 upper limits (ULs) in both laboratories were statistically higher in comparison to those provided in the IFU and in
both Russian studies, where direct and indirect techniques were applied (16.99, 16.02 vs. 14.41, 14.2, 14.16: r<0.001).
Low limit (LL) was comparable to the Russian study (direct method) and was higher than those provided in the IFU and
in the Russian indirect study (8.35, 8.24 vs 8.4 and 7.8, 6.3) [3-5].
RIs for FT3 (pmol/L, 95%) for both labs were comparable between studies with slight non-significant difference in ULs
vs. IFU (6.5, 6.6 vs 5.99: r=0.01).
CONCLUSIONS
Current retrospective analysis is easy to use and can help laboratories assess population specific RIs of routine thyroid
assays. CLSI EP-28A3C provides guidance on how to properly verify new RIs prior to implementation. The inclusion in
CLSI guidelines of more detailed recommendations on the use of the indirect method will be useful for standardizing
this procedure.
Endocrinology
P0947
UHPLC-MS/MS METHOD FOR THE SIMULTANEOUS MEASUREMENT OF CORTISOL, CORTISONE, DEXAMETHASONE AND
ADDITIONAL EXOGENOUS CORTICOSTEROIDS IN HUMAN SERUM
F. Ponzetto 3, M. Parasiliti Caprino 3, F. Settanni 1, A. Nonnato 1, E. Ghigo 3, R. Giordano 2, G. Mengozzi 1

1
Clinical Biochemistry Laboratory, City of Health and Science University Hospital, Turin (Italy)
2
Department of Clinical and Biological Sciences, University of Turin, Turin (Italy)
3
Division of Endocrinology, Diabetes and Metabolism, Department of Medical Sciences, University of Turin, Turin (Italy)
BACKGROUND-AIM
The simultaneous measurement of dexamethasone and cortisol has proven the ability to increase the diagnostic
performance of the overnight dexamethasone-suppression test. Furthermore, the therapeutic drug monitoring of
administered corticosteroid drugs could represent a crucial tool for investigating unexpected variations of steroid
hormones’ circulating levels.
METHODS
In this work, an LC–MS/MS method for the quantification of cortisol, cortisone, dexamethasone and six additional
exogenous corticosteroids in the serum/plasma matrix was developed and validated in compliance with the ISO/
IEC requirements. To assess the efficiency of the validated method, serum samples of 75 patients undergoing the
dexamethasone-suppression test and 21 plasma samples of patients under immunosuppressive treatment with
prednisone after kidney transplant were analyzed.
RESULTS
In all dexamethasone-suppression test samples, it was possible to measure the circulating levels of cortisol, cortisone
and dexamethasone. Concentrations of the latter were for all tested patients above the proposed cut-off for the
dexamethasone-suppression test’s results (1.3 ng/mL), and the cortisol concentrations showed good correlation with
the ones measured by routine immunometric analysis, therefore confirming the screening outcome for all enrolled
patients. Prednisone was detected and quantified in all enrolled patients, confirming the use of such a corticosteroid
for immunosuppressive therapy.
CONCLUSIONS
Thanks to these two applications, we proved the overall performance of the developed LC–MS/MS method for four
target analytes. The future implementation of such an analytical tool in the clinical biochemistry laboratory’s routine
will guarantee a single and versatile tool for simultaneously monitoring dexamethasone- suppression-test results and
corticosteroid drugs’ administration.
Endocrinology
P0948
USE OF VOLUMETRIC ABSORPTIVE MICROSAMPLING (VAMS) FOR THE QUANTIFICATION OF CLINICALLY RELEVANT
ENDOGENOUS STEROID HORMONES AND PHASE II METABOLITES
F. Ponzetto 2, F. Settanni 1, A. Nonnato 1, R. Nicoli 3, E. Ghigo 2, K. Tiia 3, G. Mengozzi 1

1
Clinical Biochemistry Laboratory, City of Health and Science University Hospital, Turin (Italy)
2
Division of Endocrinology, Diabetes and Metabolism, Department of Medical Sciences, University of Turin, Turin (Italy)
3
Swiss Laboratory for Doping Analyses, Centre Hospitalier Universitaire Vaudois, Lausanne (Switzerland)
BACKGROUND-AIM
Volumetric Absorptive Microsampling (VAMS) is a recent technique used to obtain dried specimens of biological
fluids that could represent in the future a valid alternative to urine, whole blood and serum sampling for clinical
measurements. The aim of the present work was to develop a UHPLC-MS/MS method for measuring the circulating
levels of clinically relevant endogenous steroid hormones and phase II metabolites from dried blood microsamples
collected with VAMS technology.
METHODS
Chromatographic separation was obtained by optimizing a 14 min multi-step gradient and by employing fully-porous
C18 analytical column. Ammonium fluoride was added to both aqueous and organic mobile phases to reach the
high sensitivity level needed for measuring circulating levels of target steroids in 30uL dried blood microsamples
and different solvents were tested for improving the extraction of selected analytes from VAMS polymer. Developed
method was validated in accordance with ISO 17025 requirements for quantitative methods and it was finally applied
to real samples with the aim of evaluating the stability of steroidal compounds stored at different conditions for up
to 100 days.
RESULTS
The optimized chromatographic conditions allowed to efficiently separate all isomeric isobaric steroids included
in the monitored panel and guaranteed a sufficient sensitivity for detecting endogenous hormones at low pg/mL
level. The performed validation protocol, including the assessment of selectivity, matrix effects, extraction recoveries,
quantitative performance (trueness, repeatability, intermediate precision, combined uncertainty, linearity range, LOQ),
carry-over and robustness gave satisfactory results. The analysis of real samples highlighted the absence of significant
differences in measured steroid concentrations when VAMS samples were stored at room temperature, 4°C, -20°C and
-80°C for up to 100 days and subjected to up to 3 freeze and thaw cycles.
CONCLUSIONS
The developed method proved to be suitable for steroid measurement in dried blood microsamples collected on VAMS
support (30uL format). This innovative approach could represent a valid alternative to classic serum/plasma analysis in
clinical laboratories, especially in pediatric applications as a more reliable solution than DBS for quantitative purposes.
Endocrinology
P0949
HYPERANDROGENISM DUE TO OVARIAN TUMOR
S. Sánchez Asís 1, P. Argente Del Castillo Rodríguez 1, M.I. Vázquez Tavero 1, M. Santés Bertó 1, B. García García 1, A.
Barceló Bennasar 1, M.I. Pastor García 1
1
Department of Laboratory Medicine, Hospital Universitario Son Espases, Palma de Mallorca, Spain
BACKGROUND-AIM
The presence of virilization and hyperandrogenism in women may appear due to different causes, both non-tumor as
well as ovarian or adrenal tumors. Hyperandrogenism in women is characterized by the possible presence of hirsutism,
alopecia, acne and, in severe cases, virilization of the sexual organs, amenorrhea and sterility.
Androgen-producing ovarian tumors represent about 1% of all ovarian tumors and are a rare cause of
hyperandrogenism. Most tumors are benign, with some cases of low-grade malignancy, and rarely present metastatic
lesions. They are more common in menopausal women and may not appear acutely.
METHODS
Clinical case description.
RESULTS
A 58-year-old woman referred to endocrinology due to an increased testosterone concentration of 7.04 mg/mL
and dehydroepiandorsterone of 28.5 µg/dL, which points to a possible ovarian origin of hyperandrogenism. It also
highlights polyglubulia with hemoglobin concentration of 19.8 g/dL. The patient referred a hair increase during the
last 2 years, androgenic alopecia, acne and a slight clitoris increase. She was not under any treatment that caused
hirsutism. She was also being followed up for a 10 cm uterine fibroid since 2013. Abdominal CT scan showed a large
androgen producing uterine pelvic mass of 19 cm on the left side. The analysis in anatomic pathology showed a
vascularized smooth muscle stromal neoplasia (positive for actin and desmin, negative for CD10 and inhibin, estrogen
and progesterone receptor was positive, androgen receptor showed nuclear positivity in 100% of cells). Treatment with
androcur was initiated, after which testosterone levels decreased to normal values, and a hysterectomy was performed.
CONCLUSIONS
Although most causes of virilization in women are benign, it is important to make a differential diagnosis since it may
appear due to neoplastic causes that, in the case of adrenal tumors, can be malignant.
Therefore, it is important to take into account the medical history, physical examination and hormone levels, as well as
imaging and histopathological studies. In addition, management will depend on the future fertility of the patient, since
this type of neoplasm typically requires surgical resection, and adequate long-term monitoring of androgen levels to
ensure its normalization.
Endocrinology
P0950
CLINICAL AND GENETIC CHARACTERISTICS OF MEDULLARY THYROID CARCINOMA: A REVIEW OF 40 CASES
K. Sifi 3, S. Hanachi 3, K. Boudaoud 2, S. Zekri 3, K. Benmohamed 1, N. Nouri 1, K. Benembarek 3, N. Abadi 3

1
Department of Medicine, Salah Boubnider University, Constantine3 / Endocrinology department, Ibn Badis University
Hospital, Constantine Algeria. to study
2
Research laboratory in biology and molecular genetics, Department of Medicine, Salah Boubnider University,
Constantine3 / Endocrinology department, Ibn Badis University Hospital, Constantine Algeria.
3
Research laboratory of biology and molecular genetics, Department of Medicine, Salah Boubnider University
Constantine3 / Biochemistry Laboratory of Ibn Badis University Hospital of Constantine Algeria
BACKGROUND-AIM
Medullary thyroid cancer (MTC) is a neuroendocrine tumor that develops from thyroid C cells. It can occur in two forms:
true sporadic in 75% of cases or familial in about 25% of cases. It is then integrated in the multiple endocrine neoplasia
type 2 (MEN2A, MEN2B, familial medullary thyroid cancer (FMTC)).
Germline mutations in the RET proto-oncogene are well known to be the genetic cause of MTC in MEN2 and can be
identified. Familial MTC is the example of a cancer that benefits of genetic screening.
The aim of our study was:
-To study the phenotypic and evolutionary characteristics of patients with MTC in Eastern Algeria.
METHODS
This is a retrospective and prospective study of 40 MTC index cases collected in 6 years and referred by
the endocrinology department of the benbadis university hospital of Constantine. The clinical and evolutionary
characteristics of MTC were assessed.
RESULTS
We enrolled 40 patients from 25 unrelated families distributed as follows:
Nine (9) MEN2A cases, 31 cases of sporadic MTC. The diagnostic criteria of MTC were confirmed by pathological anatomy
and in some cases by genetic study.
MTC was constant in all 40 patients. The mean age of its discovery was (31.28 ±13.71) years. In one case MTC preceded
the occurrence of PHEO by 4 years, in three cases MTC and PHEO were discovered simultaneously and in 36 cases MTC
was isolated. Its diagnosis was made during a systematic screening in 31. 28% of cases and after surgery of a goiter
in 68.72% of cases.
Clinically, the thyroid nodule was present in 62.50%, lymph nodes in 20.99%, multinodular goiter in 12.51%, malignant
hypertension in 4%. Other associated signs found were: motor diarrhea + thyroid nodule in 12%, Notalgia + motor
diarrhea + HTA in 4% and flush syndrome in 4%.
Preoperatively, the calcitonin (CT) assay performed showed levels ranging from 10 and 150 pg/ml in 20 % of cases ,
above 1000 pg/ml in 8% of cases and not tested in the remaining cases. Post-operatively, the high levels were between
11 and 766 pg/ml.
RET gene mutation was found in 25% of cases (C634Y). Remission was noted in 41% of cases.
CONCLUSIONS
Our results are similar to those reported in the Literature.
Endocrinology
P0951
CORTISOL IN SEMINAL PLASMA: A NOVEL TOOL IN THE EVALUATION OF MALE REPRODUCTIVE FUNCTION
G. Salvio 2, V. Viola 3, A. Calcinari 3, M. Pavani 3, L. Babini 3, B. Niccoletti 1, A. Ciarloni 2, N. Delli Muti 2, M. Martino 2, G.
Balercia 2, G. Arnaldi 2, M. Moretti 3
1
DISCLIMO, Dipartimento di Scienze Cliniche e Molecolari,Università Politecnica delle Marche, Ancona
2
SOD Clinica di Endocrinologia e Malattie del Metabolismo,Azienda Ospedaliero Universitaria delle Marche, Ancona
3
SOD Medicina di Laboratorio, Azienda Ospedaliero Universitaria delle Marche, Ancona
BACKGROUND-AIM
Several experimental evidence on animal models suggests a potential effect of glucocorticoids (GCs) on male
reproductive function. Indeed, stress-related increase in endogenous levels of serum GC can induce apoptosis in Leydig
cells and male germ cells, and the administration of exogenous GCs can lower number ad functionality of mature
sperm cells. On the other hand, the complete absence of GCs is associated with atrophy of seminiferous tubules and
spermatogenesis arrest, suggesting the existence of a physiological range of GC levels for optimal testicular function.
The aim of the present study was to measure cortisol levels in human semen plasma and to evaluate the possible
relation between sperm cortisol levels and semen parameters.
METHODS
Semen samples were provided from 115 men referring to Endocrinology Clinic, University Hospital delle Marche,
Ancona, for fertility evaluation. After liquefaction, all sperm samples were evaluated for sperm parameters
according to the 5th edition of WHO Manual. Concentrations of cortisol in seminal plasma were measured by
electrochemiluminescence (ECLIA).
RESULTS
Patients were 35.1 ± 8,9 years old. Eighty-eight subjects (76.5%) were normozoospermic, 19 subjects (16.5%) had
oligoasthenoteratozoospermia (OAT), and 8 men (7.0%) were azoospermic. High variability in sperm cortisol levels
emerged (median 2.680, IQR 1.948-4.560 mcg/dl). We found no direct correlations between sperm cortisol and semen
parameters, but significantly (p=0.02) higher levels of sperm cortisol were found in azoospermic patients (4,500;
IQR 3,295-20,385) compared with normozoospermic men (2,670; IQR 1,987-4,695) and men with OAT (2,120; IQR
1,480-3,360). Interestingly, sperm cortisol levels were almost double in men with varicocele (16%) compared with men
without varicocele: 4,280 (3,300-6,225) versus 2,410 (1697-3,805) (p=0.042).
CONCLUSIONS
To the best of our knowledge, our study was the first to evaluate the presence of cortisol in seminal plasma and its
relationship with seminal parameters. Our results show that subjects with varicocele and azoospermia have higher
seminal cortisol levels, suggesting that cortisol might influence male reproductive function.
Endocrinology
P0952
DEVELOPMENT AND VALIDATION OF A SENSITIVE LC/MS-MS METHOD FOR ESTRADIOL AND ESTRONE DETERMINATION
ON A XEVO TQ-XS (WATERS®)
C. Le Goff 1, C. Defourny 3, J. Deleersnyder 2, G. Collard 2, N. Ferrante 2, S. Peeters 1, E. Cavalier 1

1
Department of Clinical Chemistry, University of Liège, CHU Sart-Tilman,CIRM, Liège, Belgium
2
Department of Clinical Chemistry, University of Liège, CHU Sart-Tilman,Liège, Belgium
3
Department of Clinical Chemistry, University of Liège, Liège, Belgium
BACKGROUND-AIM
Estrogens are hormones responsible for the development, maintenance of the female reproductive organs and
secondary women sexual characteristics and involved in the regulation of the menstrual cycle and the maintenance of
pregnancy. The main difficulty for the quantitative analysis of these molecules is their very low serum concentration (in
the order of pg/mL). The aim of this work was the development of a LC/MS-MS assay method to concurrently determine
the two major endogenous estrogens, estrone (E1) and 17β-estradiol (E2).
METHODS
The sample preparation procedure is based on liquid-liquid extraction and derivatization with dansyl chloride. After
chromatographic separation performed on a C18 column, the compounds were analyzed using a UPLC H-Class PLUS
coupled to XEVO TQ-XS triple-quadrupole mass spectrometer operating in multiple reaction monitoring mode (Waters).
The ionization source used is the electrospray in positive mode. The procedure was validated by testing 8 levels of
concentration (from 1 to 800 pg/mL) in quadruplicate during 3 different days. A calibration curve was prepared using 7
points and responses were determined by calculating the integrated peak area ratio between E1 and E2 to deuterated
respective compound. According to these results, the precision, the trueness, and the linearity were calculated. A
comparison was performed (n=27) between our method routinely used (QTrap 6500, Sciex) with our newly developed
method (Xevo TQ-XS).
RESULTS
The relative biases obtained were between 1.3% and 4.9% for estrone and between 0.1% and 11.5% for estradiol. The
intra-assay CV did not exceed 4.6 and 6.9% and inter-assay CV, 6.9% and 7.6% for estrone and estradiol respectively.
The measurement uncertainty values obtained were < 13.9% for estrone and < 16.2% for estradiol. Very good sensitivity
was also demonstrated, with LLOQs calculated at 3.3 pg/mL and 3.7 pg/mL for estrone and estradiol respectively. No
carry-over has been demonstrated. For the method correlation, regression curves were: Old E1 method = -0, 2655 + 1,
0157 New E1 method and Old E2 method = -0, 5051 + 1, 0379 New E2 method.
CONCLUSIONS
Our new method has been successfully validated according to all the criteria set by the European Medicines Agency
(EMA) guidelines and represents a very good alternative to the immunoassays.
Endocrinology
P0953
DIAGNOSTIC RELEVANCE OF ALDOSTERONE DETERMINATION AFTER SELECTIVE SAMPLING OF ADRENAL VEINS
L. Suárez Garzón 1, A. Babinou El Khadari 1, M.A. Hadi Barghout 1, J.M. Romero Noguera 1
1
VIRGEN DE LAS NIEVES UNIVERSITY HOSPITAL, GRANADA
BACKGROUND-AIM
The diagnosis of primary hyperaldosteronism can be complicated when it is time to demonstrate the type and location
of the adenoma and, in these cases, the laboratory is of great diagnostic relevance.
METHODS
Case report: A 55-year-old patient with refractory arterial hypertension, under study due to abdominal pain caused
by acute complicated diverticulitis. During admission, an abdominal computed tomography(CT) scan showed a 2.4 cm
adrenal incidentaloma.
RESULTS
In analytical study of hypertension Aldosterone 186.6pg/mL and Renin 0.7pg/mL, normokalemic and normal in other
parameters, suggestive of hyperaldosteronism. Saline test with Aldosterone concentration >10ng/dL confirms the
diagnosis of primary hyperaldosteronism(PHA) in the context of adrenal nodule(Conn's disease). To know the subtype,
complementary tests are requested:
Gammagraphy suggestive of adrenal hyperplasia (inconclusive result).It is decided to repeat CT showing hyperplasia
of the adrenal glands with nodulations of greater size on the left, which seem to indicate that the adenoma is in the
left adrenal gland.
Selective Venous Sampling(SVS) is permormed with these results:
Right adrenal gland: cortisol 17.8; ALDOSTERONE 2307(38-150pg/ml) and Renin 2.2pg/ml. Left adrenal gland and
central vein with cortisol, aldosterone, and renin in range.
From the laboratory we reported the location of the Aldosterone producing adenoma in the Right Adrenal gland(Conn's
syndrome), contrary to what the imaging tests suggested.
The patient finally underwent surgery to remove the adenoma and blood pressure normalized.
CONCLUSIONS
Once the clinical diagnosis of hyperaldosteronism has been established, the evaluation of the adrenal glands in imaging
studies allows the morphology and vascularization of the adrenal glands to be assessed, as well as detecting possible
nodules. However, imaging tests have limited sensitivity to distinguish the location of the adenoma. SVS of the adrenal
veins is the best option. It is a fast, safe and accessible procedure that allows differentiation between uni or bilateral
PHA, which has important transcendence in the therapeutic attitude to follow.
From the laboratory we are involved from the collection of the samples, transportation to the laboratory to their
processing and interpretation of the results.
Endocrinology
P0954
SEMI-QUANTITATIVE IODINATION MAPPING BY MASS SPECTROMETRY DISTINGUISHES NORMAL HUMAN
THYROGLOBULIN FROM THYROID CANCER DERIVED THYROGLOBULIN.
B. Netzel 1, A. Maus 1, S. Grebe 1

1
Mayo Clinic, Rochester, Minnesota, USA
BACKGROUND-AIM
Thyroglobulin (TG), is iodinated in the thyroid’s follicles, becoming the precursor of thyroid hormones, which are
released when TG relocates into thyrocytes and is proteolyzed. Measurement of TG is a pillar of monitoring patients
with thyroid cancer, but tumor-derived TG can’t be distinguished from normal TG; however, thyroid cancer does not
form normal follicles, and its TG might lack iodination. This could allow specific detection of tumor derived TG.
METHODS
TG of >95% purity was obtained from commercial sources, and the EC Joint Research Centre. Fine needle aspiration
biopsy (FNA) samples of benign and malignant biopsies were waste samples. Trypsin, chymotrypsin and Glu-C, were
used for TG digestion.
LC-MS/MS analysis was on an AB SCIEX 5600 in nanospray/nanoflow mode. Full scan data was recorded from m/z
300-2,000 in high resolution mode. Information dependent acquisition scans used up to 25 product ion scans, charge
states +2 to +5, energy spreads 30+/-15 eV. Survey data was processed in Protein Pilot, searching Uniprot/Swissprot
for all variations of TG. Peptide search mass tolerances were 5ppm. Cysteine-carbamidomethylation was treated as a
fixed modification. Oxidation, iodination, di-iodination, T3, and T4 were searched as variable modifications. Tyrosine or
histidine residues that were identified as mono-iodinated, di-iodinated or as T4/T3 were recorded. Iodinated peptide
peak intensity was compared with the matched non-iodinated peptides. The iodinated peptides with the highest
intensities were used to compare iodination of normal TG, normal FNAs and malignant FNAs.
RESULTS
The iodination map covered all known and several unknown iodination sites of human TG with distinctive differences
in intensity of iodination at different sites. This is the 1st human TG iodination map created with modern techniques.
Thyroid autoimmunity research and structure/function studies of TG should benefit from these data. Moreover, tumor-
positive FNAs showed much lower iodination than the negative FNA samples or the normal TG samples.
CONCLUSIONS
Our data suggests that TG iodination has potential diagnostic utility for FNA biopsies. With improved detection
sensitivity, serum TG measurements might also allow tumor specific TG identification.
Endocrinology
P0955
LATE-ONSET OF TYPE II GLUTARIC ACIDURIA: MULTIPLE ACYL-COA DEHYDROGENASE DEFICIENCY.
P. Lesmes García Corrales 1, E. Melguizo Madrid 1, A.I. Álvarez Ríos 1, C. Delgado Pecellín 1, J.M. Guerrero Montávez 1
1
Clinical analisys laboratory, University Hospital Virgen del Rocío, Seville.
BACKGROUND-AIM
Multiple acyl-CoA dehydrogenase deficiency (MADD) or type-II glutaric aciduria is caused by defects in the electron
transport of the mitochondrial ETF/ETF-QO complex, composed of a flavoprotein (ETF) and a dehydrogenase (ETFDH).
The deficit of any of them alters the oxidation of fatty acids and some amino acids, producing an intramitochondrial
accumulation of acyl-CoA esters and a decrease in FAD.
There are three forms of clinical presentation.
METHODS
A 16-year-old woman who attended the emergency department for symptoms of a month's evolution with dyspnea and
generalized muscle weakness due to mild exertion. Acute gastroenteritis in previous months together with sporadic
vomiting, weight loss of 10 kg and occasional dysphagia.
RESULTS
On examination, clear fatigability in the voice, marked axial and proximal weakness, normal-looking gait, and dyspnea
on mild exertion were observed. Analytically, elevated creatine kinase, hypertransamines were observed.
Given the suspicion of metabolic myopathy, additional tests are performed:
• Elevated levels of very long chain acylcarnitines in tandem masses (C14:1: 0.51 µM, C16:1: 0.72 µM,).
• 24h urine organic acids: slightly elevated excretion of hexaonylglycine, suberylglycine, isobutyrylglycine,
isovalerylglycine and ethylmalonic acid.
• In the serum and urine aminogram, 1-methyl-histidinuria is observed.
The results are compatible with MADD or deficiencies in the metabolism of its cofactor riboflavin. Muscle biopsy is
consistent with lipid myopathy. The genetic study detected mutations of the ETFDH¸ gene associated with MADD with
autosomal recessive inheritance, corroborating the diagnostic suspicion.
CONCLUSIONS
MADD is an inborn error of metabolism with a fundamentally biochemical diagnosis and non-specific laboratory
abnormalities, particularly in the late myopathic form. In addition, specific alterations are observed such as elevation
of very long chain acylcarnitines in combination with organic aciduria in periods of decompensation.
MADD is a pathology included in neonatal screening panels in Spain in the last decade. In patients with symptoms who
have not been screened due to their age, the pathology should be suspected and an acylcarnitine study performed.
Endocrinology
P0956
DEVELOPMENT OF A LIQUID CHROMATOGRAPHY - TANDEM MASS SPECTROMETRY METHOD FOR THE QUANTITATION
OF OXYTOCIN.
E. Grifnée 1, F. Blistein 3, J. Demeuse 2, L. Huyghebaert 1, P. Massonnet 1, M. Schoumacher 2, T. Dubrowski 1, S. Peeters

1
, E. Cavalier 1, C. Le Goff 1
1
Department of clinical chemistry, CHU Liège, Liège
2
Department of clinical chemistry, ULiège, Liège
3
University of Liège
BACKGROUND-AIM
Oxytocin is a 9 amino acid peptide that serves as neuromodulator in the human central nervous system. This peptide is
involved in the regulation of diverse social behaviors and contributes to positive social interaction by reducing stress
and boosting self-confidence. Patients suffering from psychiatric troubles such as depression present important deficit
in terms of social and emotional competence. A reliable and sensitive assay for oxytocin is essential to reach relevant
conclusions and to support clinical interventions. Currently, oxytocin is measured by radioimmunoassay or enzyme-
linked immunosorbent assay. These methods are lacking selectivity due to cross-reactivity with plasma proteins. The
purpose of this work is to develop a sensitive and specific method for the quantitation of oxytocin by coupling liquid
chromatography (LC) to mass spectrometry (MS).
METHODS
Plasma samples have been extracted by solid phase extraction (SPE) with an OASIS MAX 96-well elution plate,
evaporated and then reconstituted. Separation and quantification were achieved on an ACQUITY UPLC H-Class PLUS
coupled to a Xevo TQ-XS (both from Waters, Mildford, USA). Chromatographic separation was performed on an ACQUITY
UPLC BEH C18 column (100 Å, 3 µm, 2.1 mm x 100 mm) (Waters, Mildford, USA). The mobile phases composition was
water and acetonitrile both containing 0.1% formic acid.
RESULTS
First, MS scan and product ion experiments were performed to define the transitions. The source and compound
parameters were optimized by post-column flow injection. The LC parameters as well as the mobile phases were
optimized based on a flow injection analysis. Once it was done, the sample preparation was developed. To obtain an
orthogonal technique compared to the column, an Oasis MAX elution plate was selected to perform the SPE. A recovery
rate of 52 % was obtained after optimization of the wash and elution solutions. A calibration curve was realized in cow
plasma with calibrators going from 10 pg/mL to 1.103 pg/mL. At a 20 pg/mL concentration of oxytocin, the peaks were
intense enough with a good and symmetrical peak shape.
CONCLUSIONS
As the endogenous concentration of oxytocin can drop below 1 pg/mL, optimization of the sample preparation needs
to be continued to lower the limit of quantification (i.e., 20 pg/mL).
Endocrinology
P0957
TOWARDS B-CTX QUANTITATION STANDARDIZATION : FIRST STEPS OF CHARACTERIZATION
J. Demeuse 2, P. Massonnet 1, E. Grifnée 1, L. Huyghebaert 1, M. Schoumacher 1, T. Dubrowski 1, W. Determe 1, S. Peeters

1
, C. Le Goff 2, E. Cavalier 2
1
Department of Clinical Chemistry, University Hospital of Liège
2
Laboratory of Clinical Chemistry, CIRM, University of Liège
BACKGROUND-AIM
The crosslinked carboxy-terminal telopeptide of type I collagen (B-CTX) is a biomarker used for the follow-up of
osteoporotic patients. It is currently uncharacterized, making it impossible to standardise the various immunoassays
for B-CTX. Little is known about the structure of the marker, other than the fact that it is a product of type I collagen
digestion and consists of three peptides linked by a pyridinoline crosslink.
METHODS
Plasma, serum, and urine pools were processed using two distinct sample preparation protocols. In the first
sample preparation, protein precipitation was used to prepare and concentrate the pools. Using preparative liquid
chromatography, samples were then separated, and fractions were collected every 15 seconds. Following evaporation,
fractions were reconstituted with 30 uL of H2O, 0,4% FA. The other sample preparation consisted of affinity
chromatography with two antibodies directed against a well-known portion of B-CTX. After samples were purified, they
were evaporated and reconstituted with 30uL of injection solvent. Both protocols' samples were then injected into a
high resolution mass spectrometer (Synapt XS, Waters). The mass spectra we obtained were then analyzed by PLGS
and Peaks Studio for type I collagen peptides identification.
RESULTS
We were able to identify ten uncrosslinked peptides of type I collagen containing the crosslink site so far but
additional database searches are currently underway to identify crosslinked peptides. These findings would lead to a
full characterization of B-CTX.
CONCLUSIONS
The successful development of our workflow led to the identification of several candidates. Confirmation of these
candidates needs further investigation.
Endocrinology
P0958
PEDIATRIC REFERENCE INTERVALS FOR FREE T4 AND FREE T3 WERE ESTABLISHED USING INDIRECT SAMPLING
TECHNIQUE IN POLAND
J. Brąszkiewicz 5, M. Szczur 5, E. Dworniczak 4, E. Zapała 4, M. Nowacka 3, L. Cesili 1, A. Ruzhanskaya 2

1
Beckman Coulter Inc., Chaska, USA
2
Beckman Coulter LLC, Moscow, Russia
3
Beckman Coulter LLC, Warsaw, Poland
4
DILAB Medical Diagnostic Nisko, Poland
5
VITALABO Medical Diagnostic Laboratories, Bydgoszcz, Poland
BACKGROUND-AIM
According to the CLSI/IFCC Document C28-A3, deriving RIs is the responsibility of each clinical diagnostic laboratory.
Most RIs provided by assay reagent manufacturers and other studies are established based on different populations,
including American and Canadian, and might not match to the Polish population. The aim of the current study was to
derive RIs for FT3 and FT4 for pediatric individuals in two big laboratory chains in Poland (Dilab, Lab1 and Vitalabo,
Lab2), using the indirect technique and to compare them to RIs established during the Canadian Laboratory Initiative
on Pediatric Reference Intervals (CALIPER) using the direct method. In both studies results were obtained using the
same analyzer, UniCel DxI 800 (Beckman Coulter, Inc.).
METHODS
Based on publications, the inclusion criteria of the retrospective study were defined as the presence in each group of
> 1000 results of FT4 and/or FT3, obtained from non-repeatable pediatrics patients 1-<19 yo of non-endocrinological
departments with TSH within the respective RI used in laboratory. Around 15.000 results were tested for TSH, Free T3,
and/or Free T4 in total and were determined to have normal TSH values as defined by a TSH reference interval of 0.38–
5.33 µIU/mL. 95%, 97.5%, and 99% RIs were calculated for Access Free T4 and Access Free T3 (Beckman Coulter Inc.).
For calculation the JMP statistical software was used.
RESULTS
RIs for FT3 and FT4 for Lab1 and Lab2 (pmol/L, 95%) had no obvious statistical differences and were combined with
next dividing by sex and age. For FT3 the most prominent differences in comparison with CALIPER were obtained in
LLs and ULs in 1-<15 yo group for males: 5.2-8.5 vs 3.98-6.19 and in ULs for females: 4.8–8.9 vs 3.98-6.19. In 15-<19 yo
group ULs were significantly higher in both sexes: 4.8–8.0 vs 3.81-5.67 and 4.3–7.4 vs 3.47-5.31 for males and females
respectively. For FT4 no obvious differences were obtained between sexes in both studies for individuals 3-<19 yo, but
the difference in ULs between Polish and CALIPER studies was prominent in both sexes: 8.4–15.5 vs 7.85-13.64 for
males and 8.3–15.1 vs 7.85-13.64 for females.
CONCLUSIONS
We have obtained prominent difference between FT3 and FT4 RIs with CALIPER study, which was targeted on the
Canadian pediatric population.
Endocrinology
P0959
CONFIDENCE IN YOUR CALIBRATORS: ASSESSMENT OF MASSTRAKTM ENDOCRINE CALIBRATOR AND CONTROL SETS
L. Calton 1, D. Foley 1, G. Hammond 1, N. Breen 4, P. Rossiter 4, G. Bosch 2, S. Donzelli 3

1
Waters Corporation, Stamford Avenue, Altrincham Road, Wilmslow, UK
2
Waters S.A.S., St Quentin, En Yvelines Cedex, France
3
Waters SPA, Sesto San Giovanni, Milan, Italy
4
Waters Technologies, Wexford, Ireland
BACKGROUND-AIM
The analytical performance of the Waters MassTrak Endocrine Calibrator and Quality Control Sets (IVD) were evaluated
using a laboratory developed LC-MS/MS method, to analyze twelve steroid hormones in a single run.
METHODS
The MassTrak Endocrine Calibrator and Quality Control Sets were used to quantify testosterone, androstenedione,
17-OHP, 11-deoxycorticosterone, DHEAS, cortisol, 11-deoxycortisol, 21-deoxycortisol, corticosterone, DHT, DHEA and
progesterone in human serum.
Internal standards were added to serum samples and precipitated with methanol, followed by dilution with water
and centrifugation. Sample preparation was completed using an SPE protocol and Waters Oasis™ MAX SPE µElution
Plate prior to injection onto the LC-MS/MS system. Using an ACQUITY™ UPLC™ I-Class FTN System, samples were
separated on a 2.1mm x 100mm CORTECS™ C8 2.6µm Column using a water/methanol/ammonium fluoride gradient
and quantified with a Xevo™ TQ-S micro.
RESULTS
The linearity and analytical sensitivity of the Calibrator Set across the respective ranges for the steroids was
successfully demonstrated. Total precision and repeatability across the steroids (3 pools, 5 replicates, 5 days; n=25)
was < 15% CV. EQA material was evaluated to demonstrate accuracy for seven of the steroids. All measured values, using
the commercial calibrators, were within 10% of the calculated LC-MS/MS mean concentrations for participating labs.
CONCLUSIONS
The Waters MassTrak Endocrine Steroids Calibrator and QC Sets (IVD), with the developed method described herein,
provides linearity, analytical sensitivity, precision, and accuracy. The accuracy data demonstrates confidence in the
Endocrine Steroid Calibrators, providing a key piece to the solution to steroid hormone standardization (ISO 15189)
and harmonization challenges in clinical laboratories.
This method is an example of an application using the instrumentation, software and consumables described in
this document. This method has not been cleared by any regulatory entity for diagnostic purposes. The end user is
responsible for completion of the method development and validation. MassTrak Immunosuppressant Calibrator and
Quality Control Sets are not available for sale in all countries.
Endocrinology
P0960
SALIVARY CORTISOL MEASURED WITH A MASSCHROM® KIT BY LIQUID CHROMATOGRAPHY–TANDEM MASS
SPECTROMETRY: METHOD VALIDATION IN THE SPIRIT OF IVDR
C. Le Goff 2, S. Peeters 1, N. Vanden Broeck 1, A. Gendebien 1, C. Nix 1, M. Rechchad 1, E. Cavalier 2

1
Department of Clinical Chemistry, University Hospital of Liège, CHU Sart-Tilman,Liège, Belgium
2
Department of Clinical Chemistry, University of Liège, CHU Sart-Tilman,CIRM, Liège, Belgium
BACKGROUND-AIM
Cortisol measurement is useful in evaluation of Cushing syndrome, adrenal insufficiency, mineralocorticoid excess and
congenital adrenal hyperplasia. Saliva is a very useful matrix for the cortisol measurement because the collection is
easy for the patient. We validated the MassChrom kit labelled IVD for the cortisol measurement in saliva developed
by Chromsystems®.
METHODS
The MassChrom kit for the saliva cortisol measurement (Chromsystems®) was validated on our Qtrap5500 (Sciex ®).
For sample preparation, deuterium labelled cortisol was added as internal standard and a filtration was performed.
The filtrate is collected and decanted into vials with micro-insert for injection and analyzed by LCMS/MS. For the
validation protocol, we followed the EMA guidelines requirements. We tested 7 levels of concentration (water spiked
with known amounts of cortisol) (0.5 -50 ng/mL) in quadruplicate during 5 days. We used the e-Noval software to
process all the data obtained from validation, allowing to study: trueness, imprecision, linearity, uncertainty and limit
of quantification (LOQ). We carried out also matrix effect, linearity of dilution, carry-over and extraction efficiency. We
compared also 20 patient samples to check the clinical concordance of the results between our routine method (an in-
house method) and the Chromsystems one.
RESULTS
The compounds showed a good linearity for the range tested. The limit of quantification was 0.5 µg/L. The intra-assay
and inter-assay CV didn’t exceed 6.9% and 8.4%. The relative biases obtained were between -0.25 % and 7.8% .The
measurement uncertainty values obtained were < 18.3%.For the linearity of dilution, the dilution up to 4 meets the
NCCLS recommendations. No carry-over was demonstrated. Mean extraction efficiency was above 53% while matrix
factor was above 90%. Clinically, we obtained 100% of concordance for the patient samples compared with the new
method.
CONCLUSIONS
The MassChrom kit for the saliva cortisol measurement was successfully validated on our Qtrap 5500 ( Sciex). The use
of IVD(R) labelled kit will help us in the future to meet the standards required by the new IVDR regulations. In mass
spectrometry, users often develop their own methods but it will no longer be possible to use them if a kit exists on
the market with the same characteristics.
Endocrinology
P0961
IS SERUM SELENIUM STATUS RELATED TO GESTATIONAL DIABETES MELLITUS IN PREGNANT WOMEN AT SECOND
TRIMESTER?
M.P. Genova 2, B. Atanasova 2, I. Ivanova 3, K. Todorova 5, K. Tosheva 4, T. Daskalova 1

1
Clinic of obstetrics and gynecology, VitaHospital, Sofia
2
Department of Clinical Laboratory, Faculty of Medicine, University Hospital “Alexander”, Medical University of Sofia,
Bulgaria
3
Department of Clinical Laboratory, St.Ivan Rilski University Hospital, Sofia, Bulgaria
4
MedicalCenter “Pentagram”
5
University Clinic of Endocrinology, MedicalUniversity of Pleven, Bulgaria
BACKGROUND-AIM
Selenium (Se) is an essential trace element which is of interest with a focus to gestational diabetes mellitus (GDM).
The evidence about the relationship between Se and GDM is controversial. The aim of the study is to investigate the
link between maternal serum Se levels with GDM at late second trimester.
METHODS
This case-control study comprises 123 pregnant women aged 18 – 40 years and in the late second trimester. Two groups
are divided-pregnant women with GDM (n=54) and pregnant women with normal glucose tolerance (NGT) (n=69). All
patients were assessed with 2-h 75-g oral glucose tolerance test (OGTT). Information of maternal social demographic
characteristics is collected using questionnaires. Blood glucose is measured by amperometric method (Biosen C-line,
Germany). Serum Se levels are analyzed by Graphite Furnace Atomic Absorption (GFAA) spectrophotometry (Perkin
Elmer Zeeman 5000, USA) with Zeeman correction of background spectral absorption.
RESULTS
The results are as follows–GDM vs. NGT pregnant women: fasting blood glucose 5.3 (5.1-5.4) vs. 4.4 (4.1-4.7) mmol/
L, p<0.001; 1-h OGTT 8.3±2.3 vs. 6.7±1.5 mmol/L, p<0.001; 2-h OGTT 6.0 (4.7-7.6) mmol/L vs. 5.0 (4.4-6.1) mmol/L,
p=0.02 and Se 55.9±13.8 µg/L vs. 64.3±11.0 µg/L, p<0.001. By using ROC curve, the best cut-off point for prediction
of GDM is obtained 50.095 µg/L Se. Sensitivity and specificity of this point are 44.4% and 94.2%, respectively, being
statistically significant. The area under the curve is AUC=0.672, p<0.001. According to the logistic regression analysis,
every 1-SD reduction in serum Se is associated with a 5.4% increased risk of developing of GDM (HR 0.946, 95% CI:
0.916-0.976; p<0.001).
CONCLUSIONS
Conclusion: Serum Se levels are lower in GDM pregnant women in the setting of hyperglycemia and insulin resistance in
comparison to control group. Our study points Se as a risk factor for development of GDM at the late second trimester.
Endocrinology
P0962
OBTAINING REFERENCE VALUES FOR HOMA-IR IN AN ADULT POPULATION OF OUR HEALTH AREA
M.I. Díaz López 1, M.D. López Abellán 1, S. Attaibi 1, Á. Puche Candel 1, A. Fernández Reina 1, L. García De Guadiana-
Romualdo 1, M.T. Orgaz Morales 1, J.R. Martínez Inglés 1, L. Rivera Soria 1, C. Nieto Sánchez 1
1
Laboratory Medicine Department, Hospital Universitario Santa Lucia, Cartagena, Spain
BACKGROUND-AIM
Insulin resistance (IR) is a inability in the biological activity of insulin and it is characterized by the need for a high level
of insulin to maintain plasma homeostasis. It is associated to obesity, type 2 diabetes mellitus and other cardiovascular
diseases, now know as metabolic syndrome. The mathematical model proposed by Matthews et al, "Homeostasis Model
Assessment of Insulin Resistance" (HOMA-IR) allows estimating it with fasting glucose and insulin levels.
Given the high rate of obesity in the current population and the fact that our laboratory did not have a reference
interval for HOMA-IR, we have considered to establish values based on our population with the methodology used in
our laboratory.
METHODS
128 apparently healthy adult patients from primary care were selected. Inclusion criteria were: plasma glucose<90mg/
dL, triglycerides<200mg/dL, total cholesterol<200mg/dL, alanine transaminase<33U/L, glomerular filtration rate (CKD-
EPI)>90ml/min/1.73m2. Exclusion criteria were: patients treated with hypolipidemic or antidiabetic drugs, patients
with obesity (Body Mass Index>30kg/m2) and diagnosed with arterial hypertension.
HOMA-IR was calculated using the formula published by Matthews: [glucose(mg/dL) x insulin(mU/mL)]/405. Glucose
was determined by hexokinase method and insulin by electrochemiluminescense (both on the Cobas 8000, Roche
Diagnostics) following the manufacturer's recommendations.
The data were analyzed following Reed's proposal for the detection of atypical results, normality testing by the
Kolmogorov-Smirnov test and determination of the reference interval by the Laplace-Gauss parametric method
recommended by Clinical and Laboratory Standards Institute, using the software SPSS.
RESULTS
The average age of the population was 40 years old with an age range of 18-65 years and 70 (54.7%) of them being
male. The lower reference limit was 0.24 (CI90%:0.09-0.39) and the upper reference limit was 2.66 (CI90%:2.51-2.81).
CONCLUSIONS
The results obtained allow us to establish a reference interval between 0.24-2.66. This interval is similar to others
obtained in studies published in the literature. Thus, a HOMA-IR value higher than 2.66 in our area should be taken
into account to evaluate IR.
Endocrinology
P0963
TODAY’S ROUTINE TESTOSTERONE IMMUNOASSAYS ARE UNSUITABLE TO LOWER THE GENERAL CASTRATION CUT-OFF
RECOMMENDATION TO < 0.7 NMOL/L (20 NG/DL)
H. Van Rossum 1, L. Van Winden 1, A. Bergman 1, H. Van Der Poel 1

1
The Netherlands Cancer Institute, Amsterdam, The Netherlands
BACKGROUND-AIM
Testosterone measurements are essential in the management of prostate cancer patients receiving castration and
androgen deprivation therapy. There has been an ongoing discussion on the testosterone castration cut-off, primarily
focusing on large cohort studies not specifying the testosterone measurement systems or studies using individual
testosterone measurement systems.
METHODS
This study is based on a post-hoc analysis of a testosterone method comparison study of a cohort of 120 castrated
prostate cancer patients. It investigates the suitability of general, measurement system independent, castration cut-
offs as recommended by all clinical guidelines.
RESULTS
We show that the four commonly applied testosterone immunoassays, are unsuitable to support lowering of the
castration cut-off to 0.7 nmol/L (20 ng/dL), since in 4.2 to 29.2% of all castrated patients testosterone levels are falsely
quantified to be higher than this castration cut-off, depending on the testosterone immunoassay used. Also, for the
Beckman immunoassay when using a 1.0 nmol/L (30ng/dL) cut-off, 13.3% of the results are falsely quantified to be
higher than this cut-off.
CONCLUSIONS
These results suggest that the measurement systems used in todays practice do not support lowering the testosterone
castration cut-off to 0.7 nmol/L. Furthermore, a more local, immunoassay dependent testosterone castration cut-off
should be considered.
Endocrinology
P0964
AMIODARONE-INDUCIDED THYROTOXICOSIS. A CASE REPORT
C. Burruezo Rodríguez 1, G. García De La Rosa 1, M.C. Martín Fernández De Basoa 1, M.T. Concepción Masip 1
1
Department of Clinical Analysis Laboratory, Hospital Universitario Nuestra Señora de Candelaria
BACKGROUND-AIM
Amiodarone is a potent antiarrhythmic indicated for the treatment of tachyarrhythmias. Due to its high iodine content
and structure similar to thyroxine (T4) and triiodothyronine (T3), it can cause abnormalities in thyroid function.
Amiodarone-induced thyrotoxicosis (AIT) is the most frequent abnormality and it can be of two types. Type I usually
occurs in patients with underlying thyroid pathology, in which excessive iodine intake causes a high synthesis of thyroid
hormones (TH), and in type II there is no de novo synthesis, but there is a high release of preformed hormones because
of cytotoxicity on the gland. In the same subject, both mechanisms may be present.
METHODS
A 75-year-old woman undergoing treatment with amiodarone for atrial fibrillation (AF), among other pathologies, who
attended the emergency room for non-ST-segment elevation acute coronary syndrome (NSTE-ACS) Killip II.
RESULTS
In the analytical study on admission, an undetectable TSH <0.08 µIU/ml (0.35-4.94) was detected together with an
elevation of free T4 (FT4) 3.58 ng/dl (0.70-1.48 ). AIT was diagnosed, treatment with amiodarone was suspended and
the antithyroid drug carbimazole and corticosteroids were prescribed.
Antithyroid antibodies (TSI) were requested with a positive result (1.6 µIU/ml), which is why it was associated with type
I AIT. After treatment, TH levels began to stabilize after 50 days, but given the poor evolution of the patient due to
her underlying pathology, she finally died.
CONCLUSIONS
In AIT, the stabilization of TH values is not rapid after stopping treatment, since the high lipophilicity of amiodarone
allows it to accumulate in adipose tissue from where it is slowly released. Most cases respond to drug treatment;
however, in cases with poor evolution, thyroidectomy may be necessary.
In this case, the patient responded to treatment. The role of the clinical laboratory is crucial in the diagnosis of this
adverse reaction and in the assessment of the efficacy of therapeutic measures. Serial determination of TH levels
should be recommended in all patients treated with amiodarone.
Endocrinology
P0965
GLYCOSYLATION DEFECT IN A PATIENT WITH A NEURODEVELOPMENTAL DISORDER.
L. Garre Morata 1, C. Aldana GarzÓn 1, A. Perojil Jimenez 1, T. De Haro MuÑoz 1

1
HOSPITAL UNIVERSITARIO CLÍNICO SAN CECILIO
BACKGROUND-AIM
An 11-year-old patient with intellectual disability, learning difficulties, attention deficit hyperactivity disorder and a
recognized handicap of 45%.
After successive consultations where there was no evidence of progress in the level of learning and with the aim of
identifying the intellectual disability, the Pediatric Neurology consultation decided to start a laboratory study.
METHODS
Laboratory studies for patients with neurodevelopmental diseases are complex and range from basic biochemical tests
to genetic tests.
RESULTS
•Kidney, liver and thyroid function: Normal.
•Lipid profile: Decreased.
•Acylcarnitines:, organic acid profile, blood amino acid profile and urinary oligosaccharide: Normal.
•Dimethylene ethylene blue (DMB) test for mucopolysaccharidosis screening: Normal.
•Urinary amino acid profile: Mild elevation of different amino acids without a defined pathological pattern.
•Sialotransferrins: A sialotransferrin pattern characteristic of type I deficiencies was obtained.
Congenital Deficiencies of Glycosylation (CDG):
Asialotransferrin: 0% (0%).
Monosialotransferrin: 0% (0%).
Disialotransferrin: 19,5 % (0,7- 3%).
Trisialotransferrin: 3,4 % (1,7-9 %).
Tetrasialotransferrin: 73,9 % (60-85%).
Pentasialotransferrin: 3,2 % (10,3-25%).
Subsequently, a molecular study of the genes involved in CDG was performed, which confirmed that the patient was a
heterozygous carrier of two variants of uncertain clinical significance in the ALG12 gene.
The presence of changes in this gene is associated with an autosomal recessive pattern of inheritance of congenital
disorder of glycosylation type 1g.
Glycosylation deficiency type 1g (MIM607143) is a congenital disorder of N-glycosylation characterized by
facial dysmorphism, developmental delay, cardiac abnormalities, progressive microcephaly, among others. After
confirmation of the diagnosis, the patient was referred to different services (ophthalmology, cardiology) to study
possible problems associated with the diagnosis.
CONCLUSIONS
The exhaustive study carried out by the Laboratory of Metabolopathies allowed the detection of a pattern of
sialotransferrins suggestive of this inborn error of metabolism. This was essential for a correct diagnostic orientation,
with shorter delay times and subsequent confirmation by NGS.
Endocrinology
P0966
STUDY ON EVALUATING THE TECHNICAL ACCURACY OF GLUCOMETERS IN MEDICAL WARDS AT A TERTIARY HOSPITAL
IN SRI LANKA
P. Samarakoon 1, S. Gunawardena 1, T. Inthujah 1, G. Dissanayake 1, M. Balasooriya 1, S. Sayanthan 1, Z. Thaneefa 1, T.

Prashanthan 1, G. Katulanda 1
1
Department of Chemical Pathology, National Hospital Sri Lanka
BACKGROUND-AIM
Glucometers are important point of care devices that are being used in a variety of health care settings. Most dynamic
function tests (insulin tolerance test, glucagon stimulation test and prolong supervised fast) need obtaining samples
for hormones at the point of hypoglycaemia.
Recently we came across a few instances where large analytical discrepancies were noted between capillary and venous
glucose values during dynamic function tests which became an issue in interpreting results as the cutoffs are set to
be interpreted at defined plasma glucose values.
So the aim was to, assess the accuracy of glucometers used for glucose monitoring during dynamic function tests in
the wards.
METHODS
Five glucose samples (64, 87, 100, 189, 400 mg/dL) were prepared using pooled plasma and analyzed using glucometers
in 12 medical wards at NHSL. Bias was calculated at each concentration for all glucometers. Results interpreted against
ISO 15197:2013 guideline.
RESULTS
6/12 (50%) glucometers showed an accurate result at concentrations below 65 mg/dL while 10/12 (83%) were accurate
at concentrations >400 mg/dL. 3/ 12 glucometers were accurate at all concentrations according to the guideline. 6/12
showed a negative bias at <65mg/dL.
CONCLUSIONS
Negative bias in the results can lead to false diagnosis of hypoglycaemia, which can lead to collection of hormones
samples at higher concentrations of glucose which makes the test invalid. It is very important to use glucometers with
minimal bias during dynamic function tests to reduce the wastage of reagents and to minimize the cost in a clinical
laboratory.
Endocrinology
P0967
GENETIC SHORT STATURE: A CASE REPORT WITH WEILL-MARCHESANI SYNDROME AND GHRELIN-RECEPTOR GENE
VARIANT
G.A. Marina 2, D.T. Noelia 2, V.P. Maria Irene 5, C.E. Berta 4, C.M. Ariadna 1, H.A. Sara 1, Y.F. Diego 1, F.C. Roser 2, B. Betina 3
1
Department of Pediatric Endocrinology, Vall d’Hebron University Hospital, Barcelona, Spain
2
Department of Biochemistry, Vall d’Hebron University Hospital, Barcelona, Spain
3
Department of Endocrinology, Vall d’Hebron University Hospital, Barcelona, Spain
4
Department of Clinical and Molecular Genetics, Vall d’Hebron University Hospital, Barcelona, Spain.
5
Department of Clinical Genetics and Minority Diseases, Vall d’Hebron University Hospital, Barcelona, Spain.
BACKGROUND-AIM
Familial clinical history of short stature or syndromic short stature should be studied for genetic defect.
METHODS
An 18-year-old female patient with short stature and further clinical manifestation genetic testing including whole
exome sequencing was indicated. Familial segregation studies, basal growth hormone (GH) and insulin-like growth
factor-1 (IGF-1), RNA studies and functional tests aided in the interpretation of the genetics results.
RESULTS
Physical examination showed a harmonic short stature (height 131 cm [p3], weight 44.6 kg [p3]), relative macrocephaly
with a broad forehead and clinodactyly of the fifth fingers. She had eye disturbances consisting of shallow anterior
chamber, lens subluxation, spherophakia and high myopia. Hormonal profile was normal for thyroid test, sexual
hormones and adrenal axis. GH was detectable (0.19 ng/mL) but IGF-1 was low (-2.8 SDS). GH and cortisol stimulation
test with insulin showed a correct secretion of both hormones. The anteroposterior radiograph of the wrist showed
bone age of 18 years with slightly short metacarpals and short fingers. The pituitary Magnetic Resonance Imaging
(MRI) had no relevant findings. Microarray analysis was normal. Whole-exome sequencing confirmed the presence
of a homozygosis variant of unknown clinical significance (VUS) in ghrelin receptor (GHSR) and the presence of a
homozygous likely-pathogenic variant in ADAMTS17. In order to investigate the implication of the variants found in
GHSR, macimorelin test was performed. No endogenous GH secretion was detected after macimorelin acetate. And to
further determine the pathogenicity of the ADAMTS17 variant RNA testing was also performed. Familial segregation
and functional studies were key to conclude that the phenotype observed in the patient was due to a dual genetic
diagnosis.
CONCLUSIONS
The identification of a homozygous variant in ADAMTS17 confirmed the diagnosis of Weill-Marchesani syndrome, a
rare genetic disorder characterized by short stature, brachydactyly, joint stiffness and characteristic eye abnormalities.
The clinical significance of the homozygous variant found in GHSR was confirmed using a ghrelin analog. Both variants
contribute to the phenotype observed in the patient. Diagnosis in this patient was relevant to better follow up and
familial genetic counselling.
Endocrinology
P0968
UTILITY OF 24-HOUR STEROID PHARMACOKINETIC IN CLASSICAL CONGENITAL ADRENAL HYPERPLASIA
N. Díaz-Troyano 1, M. Giralt-Arnaiz 1, C. Aguilar-Riera 2, D. Yeste-Fernández 2, R. Ferrer-Costa 1, M. Clemente-León 2

1
Department of Biochemistry, Vall d’Hebron University Hospital, Barcelona, Spain
2
Department of Pediatric Endocrinology, Vall d’Hebron University Hospital, Barcelona, Spain
BACKGROUND-AIM
The 24-hour steroids pharmacokinetic are used to assess the circadian rhythm induced in patients with classical
Congenital Adrenal Hyperplasia (CAH) on corticosteroid therapy. The aim is to reduce adrenal androgen production,
but this does not mean that the cortisol (COR) replacement is adequate. For this reason, there are studies that
propose to use COR as a biomarker; if the circadian rhythm is regulated correctly, it will decrease the concentration
of corticotrophin (ACTH) and as a consequence also the concentration of androgens avoiding both overtreatment and
insufficient dosing.
METHODS
A 9-year-old female with classical CAH due to a 11-beta-hydroxylase enzyme deficiency. As a result, there is
accumulation of 11-deoxycortisol without activation of the renin-angiotensin-aldosterone system. Laboratory analysis
results: plasma renin activity <0.1 ng/mL/h (0.2–5.7), 17-OH-Progesterone 5.1 ng/mL (<3), ACTH 290.7 pg/mL (4.7–
48.8), androstenedione (AND) 8.5 ng/mL (0.1–2.5), and testosterone 98 ng/dL (14–76).
RESULTS
The patient took 15.2 mg/m2 hydrocortisone per day (4.5 mg at 1:30h, 4 mg at 7:30h and 3.5 mg at 13:00h and 19:30h).
To find out whether the schedule mimics the circadian rhythm of the corticotropic axis, the 24-hour pharmacokinetic
were performed during 3 days. The results obtained shown that the dose of 4 mg at 7.30h was insufficient because
the levels of AND rose to 1.3 ng/mL. Raising the dose of hydrocortisone to 4.5 mg, lower AND concentrations were
achieved (0.6 ng/mL). In addition, the dose was decreased to 3 mg at 19.30h since AND values did not increase (0.4
ng/mL) and circadian rhythm of COR was maintained.
CONCLUSIONS
After 24-hour pharmacokinetic is decided to keep the same dose of hydrocortisone but distributed differently: 4.5
mg at 1:30h and 7:30h, 3.5 mg at 13:00h and 3 mg at 19:30h. In practice, dose adjustment may be challenging due
to the variability of paediatric patients on the same dose of hydrocortisone, maybe due to different metabolism by
the classical and alternative pathway. The 24h pharmacokinetics is a complex procedure that requires in-patient stay
and multiple blood extractions. It may help dose adjustment in selected patients, although it needs to be repeated
as these children grow up.
Endocrinology
P0969
ELEVATED URINE NOREPINEFRINE LEVEL AND ALCOHOL ABUSE: A CASE REPORT
J. Intra 1, S. Ippolito 1, F. Lorenzini 1, A. Mauro 1, M.C. Mazzitello 1, S. Melzi 1, A. Cappellani 1, F. Cappellini 1, M. Casati 1
1
Clinical Chemistry Laboratory, Fondazione IRCCS San Gerardo dei Tintori, Monza, Italy
BACKGROUND-AIM
Alcohol abuse is associated with over 60 chronic diseases, including diabetes, hypertension, cirrhosis of the liver, breast
tumor, and early mortality.
METHODS
Here, we reported a case of elevated urine norepinephrine in a 56-years old subject with diagnosis of hypertension
treated with antihypertensive medications.
RESULTS
In regular check-up, complete blood count revealed normal values: white blood cell count 6.5 x 109/L (reference range
(RR): 4.0-11.0 x 109/L), red blood cell count 4.69 1012/L (RR: 4.50-6.00 x 1012/L), hemoglobin 154 g/L (RR: 120-160 g/L),
and platelet count 171 x 109/L (RR: 140–440 x 109/L). Biochemical analysis returned the following results: creatinine
79.2 µmol/L (RR: 61.6-105.6 µmol/L), serum sodium 144 mmol/L, serum potassium 3.9 mmol/L, and a normal value of
thyroid stimulating hormone, 2.72 uUI/mL (RR: 0.27-4.20 uUI/mL). Assessment of urinary catecholamines resulted in
a normal value of epinephrine 6.5 µg/24h (RR: 0.0-20.0 µg/24h) and an elevated value of norepinephrine 254.3 µg/24h
(RR: 15.0-80.0 µg/24h). After 2 weeks, urinary epinephrine was 6.0 µg/24h, while norepinephrine 192.2 µg/24h. Urinary
vanillylmandelic acid, urinary metanephrines, urinary, serum and salivary cortisol, plasma renin and aldosterone were
in reference intervals. A more in-depth investigation revealed that the patient had a history of severe alcohol abuse.
CONCLUSIONS
Reviewing the literature, a chronic use of drugs and alcohol has effects on the noradrenergic neurotransmitter system.
Alcohol abuse seems to increase norepinephrine turnover and release, and to downregulate the α2-adrenergic receptors
expression. However, the effects of alcohol are still in debate, most probably due to the variability of dose, length of
exposure, and reduction of alcohol intake. Acute alcohol ingestion in humans increases norepinephrine levels, while in
rodents it is not clearly demonstrated. Conversely, it is unclear if chronic alcohol administration enhances or suppresses
noradrenergic signaling. Further research is needed, urinary cathecolamines might be used as biological markers of
alcohol abuse to understand the relationship between this pathological disorder and neuroendocrine system, and to
aid clinicians in alcohol use detection.
Endocrinology
P0970
VALIDATION OF A HIGHLY SENSITIVE IN-HOUSE PLASMATIC CATECHOLAMINES LC-MS/MS METHOD
C. Le Goff 1, S. Peeters 2, M. Rechchad 2, J. Deleersnyder 2, E. Cavalier 1

1
Clinical Chemistry Department, University Hospital of Liege, CIRM, Liege, Belgium
2
Clinical Chemistry Department, University Hospital of Liege, Liege, Belgium
BACKGROUND-AIM
The determination of epinephrine (E), norepinephrine (N), dopamine (D) help to diagnose pheochromocytoma, a rare
but potentially fatal tumor arising primarily from the chromaffin cells of the adrenal medulla. However, it is not easy
to measure due to the low concentrations in plasma. The aim of this work was to validate an in house method for the
determination of plasmatic catecholamines.
METHODS
Plasma samples were extracted by solid phase extraction and analyzed by an in sample ion-pairing chromatography
LC-MS/MS on a ACQUITY UPLC H-Class PLUS (Waters, Mildford, USA) coupled to a Xevo TQ-XS (Waters, Mildford,
USA). Chromatographic separation was performed on a Kinetex Biphenyl column (2.6 µm, 2.1 mm x 100 mm) from
Phenomenex (Torrance, California, USA). The mobile phases composition was water and methanol both containing
0.1% formic acid.
Validation of the method was realized according the European Medicines Agency (EMA) guidelines. We determined the
repeatability, reproducibility, uncertainty, linearity and LOQ on free catecholamine plasma spiked with different known
concentrations of the different compounds (E, N, D). We carried out also matrix effect, linearity of dilution, carry-over
and recovery. The validation concentration range was 4.2-2083 µg/L for the 3 compounds. Each level was analysed in
fourfold in 5 run. A calibration curve (8 points) was systematically carried out with each run validation.
RESULTS
The compounds showed a good linearity for the range tested. The LOQ were: 37, 23.5 and 51.9 ng/L for E, N, D
respectively. The repeatability did not exceed 6.8 % for E, 4.7% for N and 4.1% for D. The total precision did not
exceed 9.4%, 8.8% and 7.6% for E, N and D. The mean maximum relative bias were 6.3%, 5.9% and 7.5% for E, N and D
respectively. For the linearity of dilution, the dilution up to 10 meets the NCCLS recommendations. After experiments,
recovery rate was above 77 % while matrix factor for the analyte was above 86%. No carry-over was demonstrated.
CONCLUSIONS
Our in-house method was successfully validated It represents a convincing alternative to the HPLC method for a faster
and reliable measurement of plasmatic catecholamines. Especially as there is no commercial alternative method.
Endocrinology
P0971
VON HIPPEL-LINDAU SYNDROME: APPROACH AND CONFIRMATION OF A CLINICAL CASE.
B. Nafría Jiménez 1, A. Garrido Chércoles 1, M.J. Izquierdo Vicente 1, R. Cabezón Vicente 1, C. Lallave Hernández 1, E.
Bereciartua Urbieta 1, L. Martínez González 1, I. Pérez Casas 1
1
Clinical Biochemistry Laboratory, Donostia University Hospital, Donostia-San Sebastian, Spain
BACKGROUND-AIM
Von Hippel-Lindau syndrome (VHL) is a hereditary condition with autosomal dominant inheritance caused by a
mutation in the pVHL tumor suppressor gene (3p25.5). Its inactivation predisposes to the development of multiple
vascular tumors: ocular (retinal angiomas), Central Nervous System (hemangioblastomas), clear cell renal carcinomas,
adrenal glands (pheochromocytomas), pancreas, liver and epididymis. Despite its high penetrance, the phenotypic
expression is variable, so its diagnosis is sometimes difficult and can be delayed.
METHODS
26-year-old female attended to emergency department due to hypertensive crisis with blurred vision. She was admitted
for study and the abdominal computerized axial tomography (TAC) revealed a right adrenal mass. No family history.
RESULTS
Laboratory analysis: elevated levels of catecholamines in urine, at expense of normetanephrine (783 µg/24h VR
[0-527]). Method: High Performance Liquid Chromatography (HPLC).
•Genetic study (Next Generation Sequencing): Identification of the mutation c.562C>G (p.L188V) in the VHL gene, in
homozygosis, confirms the diagnosis of pheochromocytoma. This variant is associated more frequently with VHL type
2C (ACMG pathogenic: PM1, PM2 and PM5 moderate, PP2 and PP3 supporting).
Treatment: Exeresis of left adrenal gland tumor.
Evolution:
•VHL-2C refers to patients with isolated pheochromocytoma. However, as the patient presented a specific episode of
blurred vision, she will be monitored in Ophthalmology.
•Genetic testing was performed on the rest of relatives who were negative, except for her 1-year-old daughter, who
was heterozygous for the same mutation.
CONCLUSIONS
Since VHL is a multisystem disorder with a predisposition to develop multiple neoplasms, early detection is essential
due to the prevention studies that are carried out throughout the patient´s life. The follow-up of clinically affected
patients and heterozygous individuals must be approached from a multidisciplinary perspective in which the Clinical
Laboratory is fundamental for their monitoring with biomarkers. Likewise, the molecular study confirmed the clinical
diagnosis and permitted the identification of asymptomatic relatives, allowing their inclusion in the recommended
follow-up protocol.
Endocrinology
P0972
AUTOIMMUNE POLYGLANDULAR SYNDROME TYPE 2: A CASE REPORT THAT COULD END IN AN ENDOCRINE EMERGENCY.
B. Nafría Jiménez 1, A. Garrido Chércoles 1, M.J. Izquierdo Vicente 1, R. Cabezón Vicente 1, R. Oliveros Conejero 1, E.
Redín Sarasola 1, E. Bereciartua Urbieta 1
1
BACKGROUND-AIM
Autoimmune polyglandular syndrome (PAS) is a heterogeneous group of rare diseases characterized by the coexistence
of 2 or more immune-mediated endocrinopathies. Although they are infrequent, 4 types of PAS have been described
according to the associated pathologies. In adults, type II (Schmidt syndrome) is the most characteristic and usually
presents with autoimmune adrenal insufficiency, autoimmune thyroid disease and/or type 1 diabetes mellitus.
METHODS
45-year-old female attended to emergency department due to diarrhea and abdominal pain in last 2 days, together
with the loss of 11 kg in last 3 months. Physical examination: vitiligo lesions on feet and hands, and generalized
hyperpigmentation. Medical history: primary autoimmune hypothyroidism.
RESULTS
Laboratory results on admission: TSH 0.24 mU/L [VR 0.3-4.2], free-T4 1.03 [VR 0.85-1.85], sodium 129 mEq/L [VR
135-145], potassium 5.2 mEq/L [VR 3.3-5.1], calcium 10.4 mg/dL [VR 8.6-10.2].
Given the suspicion of primary adrenal insufficiency, its cause must be investigated and discard other autoimmune
endocrine deficits. So, hormonal and autoimmune studies was expanded:
•Corticotropin (ACTH) 1780 pg/mL [VR 10-52] and cortisol <3 µg/dL; confirming primary adrenal insufficiency.
•21-Hydroxylase antibodies 7.80 [VR index 0.0-1.0]; confirming the autoimmune etiology.
•Plasma-renin, mass: 1,027.0 mU/L;
•Serum-Aldosterone: 98 pmol/L;
•HbA1c 7.3% [VR 4.3-5.9]: related to DM-1.
•Anti-parietal cell and anti-transglutaminase antibodies (IFI): negative.
•HLA: DRB1*03:04.
PAS type II was diagnosed, with the coexistence of primary adrenal insufficiency, thyroiditis (expressed with
hypothyroidism) and DM1 as the main autoimmune endocrinopathies, together vitiligo. Hormone replacement therapy
started for glandular insufficiencies.
CONCLUSIONS
A case of PAS is presented where early detection led to adequate hormone replacement, which is fundamental,
particularly when associated with adrenal or thyroid insufficiency since they often present as an emergency with high
mortality rates. The rapid response of the Clinical Laboratory, expanding hormonal study, favoured an early diagnosis,
also entailing family follow-up due to the possibility of more subsequent endocrinological dysfunctions.
Endocrinology
P0973
GROWTH HORMONE PROVOCATION TEST IS VALUABLE TOOL FOR DIFFERENTIATION BETWEEN GROWTH HORMONE
DEFICIENCY AND IDIOPATHIC SHORT STATURE
B. Beljan 1, V. Šupak Smolčić 2, L. Bilić-Zulle 2

1
Clinical Department of Laboratory Diagnostics, Clinical Hospital Center Rijeka, Rijeka, Croatia
2
Clinical Department of Laboratory Diagnostics, Clinical Hospital Center Rijeka, Rijeka, Croatia AND Faculty of Medicine,
University of Rijeka, Rijeka, Croatia
BACKGROUND-AIM
Inadequate secretion of growth hormone (GH) from the anterior pituitary, growth hormone deficiency (GHD), can be
diagnosed by performing a GH provocation test with insulin stimulation. The aim of the study was to compare peak GH
concentration and time to peak GH concentration after insulin-induced hypoglycemia (<2.5 mmol/L) between patients
with GHD and those with idiopathic short stature (ISS). We compared baseline levels of insulin-like growth factor-1
(IGF-1) and insulin-like growth factor binding protein-3 (IGFBP-3) between GHD and ISS patients.
METHODS
Data were retrieved from the Laboratory Information System of the Clinical Department of Laboratory Diagnostics,
Clinical Hospital Center Rijeka, Croatia, from 2018 to 2022. The case-control study included 28 GHD patients (18
males) aged 147(56-181) months and 28 ISS patients (18 males) aged 144(66-193) months, matched by age. Blood
was collected before insulin injection (baseline) and after 10, 20, 30, 60, 90, and 120 minutes. GH was measured on
Roche Cobas 6000 (Roche Diagnostics, Mannheim, Germany) in all samples, and IGF-1 and IGFBP-3 were measured on
Immulite 2000XPi (Siemens, New Jersey, USA) in the baseline sample only. Glucose was measured at each time point
using the Accu-Check glucometer (Roche Diagnostics, Mannheim, Germany). The time from hypoglycemia to maximum
GH concentration was calculated for each patient and expressed as ∆t (min). Data presented as median and IQR. The
difference between GHD and ISS patients was tested with the Mann-Withney test using MedCalc software (MedCalc,
Ostend, Belgium) with significance P<0.05.
RESULTS
GHD patients had a lower peak GH concentration than ISS patients (8.38(4.61-11.50) vs 26.09(22.02-35.08) mIU/L,
P<0.001). GHD patients had lower IGF-1 concentration than ISS patients (11.8(7.8-18.1) vs 15.4(11.2-20.5) nmol/L,
P=0.032). GHD patients had lower IGFBP-3 concentrations than ISS patients (121.0(87.7-177.5) vs 159.5(137.0-198.0)
nmol/L, P=0.002). GHD patients had a prolonged ∆t compared with ISS patients (40(20-40) vs 10(0-10) min, P<0.001).
CONCLUSIONS
GHD is characterized by lower baseline IGF-1 and IGFBP-3 concentrations and lower GH concentrations after insulin
stimulation compared to ISS patients. In GDH, the time to reach maximum GH concentration after hypoglycemia is
prolonged.
Endocrinology
P0974
EVALUATION OF THE ANALYTICAL PERFORMANCE OF THYROID-STIMULATING HORMONE, FREE TRIIODOTHYRONINE,
AND FREE THYROXINE ASSAYS ON THE ATELLICA CI 1900 ANALYZER
J. Rhea-Mcmanus 1, M. Quintanilla 1, H. Zhang 1, M. Guo 1, H. Leipold 1

1
Siemens Healthcare Diagnostics Inc., Tarrytown, NY, U.S.
BACKGROUND-AIM
The Atellica® CI 1900 Analyzer is an automated, high-throughput integrated chemistry and immunoassay analyzer
employing both Atellica CH and Atellica IM assays. This study evaluated the analytical performance of the Atellica IM
Thyroid Stimulating Hormone 3 Ultra™ (TSH3-UL), Free Triiodothyronine (FT3), and Free Thyroxine (FT4) assays* on the
Atellica CI 1900 Analyzer.
METHODS
The Atellica CI 1900 TSH3-UL, FT3, and FT4 assays use the same reagents and calibrators as the Atellica IM assays.
Precision and method comparison studies were performed according to CLSI EP05-A3 and CLSI EP09-A3 using native
and contrived human serum (TSH3-UL, FT3, and FT4) and plasma (TSH3-UL) samples. One aliquot of each sample pool
was tested in duplicate in two runs per day ≥2 hours apart on each analyzer for ≥20 days. FT3 and FT4 were evaluated
with one reagent lot on two systems. TSH3-UL was evaluated with two reagent lots on two systems. For the method
comparison, individual native human serum samples were analyzed using the Atellica IM TSH3-UL, FT3, and FT4 assays
on both the Atellica IM and Atellica CI 1900 Analyzers.
RESULTS
TSH3-UL repeatability %CVs ranged from 1.1–1.7% at concentrations of 58.362 and 4.745 µIU/mL, respectively. Within-
lab CVs ranged from 1.9–3.3% at concentrations of 1.706 and 97.533 µIU/mL, respectively. Repeatability %CVs for FT3
ranged from 0.8–13.2% at concentrations of 7.44 and 1.16 pmol/L, respectively. Within-lab CVs for FT3 ranged from
1.7–16.8% at concentrations of 7.44 and 1.16 pmol/L, respectively. FT4 repeatability %CVs ranged from 1.1–2.7%, and
within-lab CVs ranged from 3.1–6.3% at concentrations of 54.31 and 4.77 pmol/L, respectively. Method comparison of
the assays on the Atellica CI 1900 Analyzer vs. the Atellica IM Analyzer showed regression slopes of 0.96 (TSH3-UL),
1.00 (FT3), and 0.97 (FT4) and intercepts of 0.001 mIU/L (TSH3-UL), 0.15 pmol/L (FT3), and 0.26 pmol/L (FT4).
CONCLUSIONS
Evaluation of the Atellica IM TSH3-UL, FT3, and FT4 assays using the Atellica CI 1900 Analyzer demonstrated acceptable
precision and equivalent performance compared to the same assays on the Atellica IM Analyzer.
*The products/features mentioned herein are not commercially available in all countries. Their future availability
cannot be guaranteed.
Endocrinology
P0975
IMPORTANCE IN REPORTING CRITICAL VALUES, A CASE OF SEVERE HYPONATREMIA.
C. Martín Tovar 1, A. Delgado Baena 1, L. Velasco Rodriguez 1, J.A. Vilchez Aguilera 1, P. Parrado Hernando 1, J.M. Villa
Suarez 1, J. Molina Santigo 1
1
Hospital San Agustín de Linares
BACKGROUND-AIM
Hyponatremia is defined by the presence of a plasma sodium concentration lower than 135 mEq/l. It is not an entity in
itself but usually appears secondary to other pathological processes. The symptomatology of hyponatremia is often
mild, the typical clinical picture produces lethargy, confusion, vomiting and appears when hyponatremia is severe
(<120-125mEq/l).
Secondary hyperparathyroidism is a reversible state related to excessive secretion of PTH by hyperplastic parathyroid
glands, mainly due to decreased transport of calcium ions into parathyroid cells. The course of the disease is often
asymptomatic and its diagnosis is mainly biochemical, on many occasions serendipitous by finding low blood ion levels.
METHODS
Description of a clinical case and review of the literature
RESULTS
An 83 year old patient who undergoes a control blood test at her primary care center, all values are within range
except sodium levels. The patient presented severe hyponatremia with a value of 118 mEq/l, so it was decided to call
her requesting physician to inform him of the critical value. A few hours later, the patient came to the emergency
room of our hospital with even more severe hyponatremia, with values of 111 mEq/l, so it was decided to admit her.
Once on the ward, a complete blood analysis was requested, where we observed that in addition to hyponatremia,
she had hypocalcemia with corrected calcium values of 6.4 mg/dl. Given the hypocalcemia from the laboratory it was
decided to extend the parathormone study with the following results: Intact parathormone: 118 pg/mL (15-65 pg/
mL);Phosphorus: 1.7 mg/dL;Magnesium: 1 mg/dL;Vitamin D: 10.1
Finally a secondary hyperparathyroidism is diagnosed.
CONCLUSIONS
When we find hyponatremia as an isolated finding and it is not expected within the clinical context of the patient, we
must confirm it with another analytical determination. In this case the rapid intervention of the laboratory in terms of
notification of the critical value and the subsequent extension of the parathormone study allowed a rapid diagnosis,
which reached the problem of origin in only two days. It is the role of the laboratory to be alert in this type of diseases
that often occur silently and therefore are underdiagnosed or take a long time to be diagnosed with the consequent
harm to the patient.
Endocrinology
P0976
EVALUATION OF NEW CUT-OFF POINTS OF THE LHRH STIMULATION TEST IN THE DIAGNOSIS OF CENTRAL PRECOCIOUS
PUBERTY IN GIRLS
E. Rufo Vicente 1, J. Rodríguez Domínguez 1, M. Murillo Vallés 2, A. Valls Llussà 2, J. Barallat Martínez De Osaba 1, O.
L'Kadiri El Amrani 1, S. Martínez Couselo 1
1
Clinical Laboratory North Metropolitan Area / Universitary Hospital Germans Trias i Pujol, Spain
2
Pediatric Endocrinology Department / Universitary Hospital Germans Trias i Pujol, Spain
BACKGROUND-AIM
Precocious puberty is defined in girls as the appearance of secondary sex characteristics before the age of 8. Most
cases correspond to forms of central precocious puberty (CPP) with premature activation of the hypothalamic-pituitary-
gonadal-axis (HHG). Gonadotropin-releasing hormone (GnRH) stimulation tests could evidence the activation of the
HHG. However, these tests are currently not standardized and there is controversy regarding the optimal peak LH
concentration or LH/FSH ratio for CPP prediction.
The aim of the study was to evaluate the usefulness of the GnRH test (LHRH) for the diagnosis of CPP and to define
its optimal diagnostic criteria with our methodology.
METHODS
Retrospective study enrolling 38 girls, aged 5-9 years, with suspected CPP. According to clinical evolution participants
were classified into progressive (PP; n=17) and non-progressive (NPP; n=21) puberty groups. LHRH test was performed
by determining serum LH, FSH, estradiol and testosterone at baseline and LH and FSH at 15; 30; 60 and 90 minutes after
an intravenous bolus of 0.1 mg LHRH. Serum hormones concentrations were measured by Architect i2000 immunoassay
(Abbott Diagnostics). Mann-Whitney test and ROC curves were calculated using MedCalc 19.6.
RESULTS
Participant’s mean age was7.6 years (IC95%: 7.3-8.0).Baseline and stimulated LH, baseline FSH, testosterone and
baseline and stimulated LH/FSH ratio were significantly higher (p<0.05) in the PP group.
The best predictor of PP was LH/FSH ratio at 60 minutes using a cut-off of 0.35 (AUC= 0.93; 95% CI: 0.88-0.99) with
88.2% sensitivity and 95.2% specificity. Stimulated LH at 60 minutes was an optimal predictor using a cut-off of 4.7
mUI/L (AUC= 0.90; 95%IC: 0.76-0.97) with 76.5% sensitivity and 90.5% specificity.Baseline LH using a cut-off of
0.13mUI/L showed an AUC=0.81 (95%IC: 0.66-0.92) with 90.5% specificity, and70.6% sensitivity. A 100% specificity
was reached by increasing the cut-off to 0.48mUI/L, with a 47.1%sensitivity.
CONCLUSIONS
LHRH test for the diagnosis of CPP could be simplified to only a baseline and an extraction 60 minutes after intravenous
LHRH injection. LHRH test would only be indicated in patients with baseline serum LH lower than 0.48mUI/L. Stimulated
LH > 4.7 mUI/L or stimulated LH/FSH ratio > 0.35 are the optimal predictors of CPP.
Endocrinology
P0977
PANHIPOPITUITARISM DUE TO METASTASIS OF NEUROENDOCRINE TUMOUR OF LUNG: A CASE REPORT.
J. Tortosa-Carreres 1, R. Andreu-Escriva 1, A. Alba-Redondo 1, M.L. Martínez-Triguero 1, B. Laiz-Marro 1

1
Clinical Chemistry Department. Hospital Universitari i Politècnic la Fe (Valencia).
BACKGROUND-AIM
Panhipopituhitarism is a rare condition characterised by the lack of all pituitary hormones. A prevalence of 4.2 cases per
100000 habitants has been reported. It can be caused by several disorders and may be life-threatening if is untreated.
METHODS
We present a case report of 58 years-old man with 9-years neuroendocrine tumour (NET) of lung with liver metastasis
and no other backgrounds. Patient was admitted in the emergency department with dizziness, sweating, blurry vision
and weakness.
RESULTS
Given the initial results of blood test (TSH: 0.09 mU/L (0.27-5.35); free T4: 0.5 ng/dL (0.93-1.7); prolactin: 0.4 (4-15.2 );
Chromogranine A: 27980 ng/mL (0-100); NSLE: 180 ng/mL(0-15)) it was decided to perform all pituitary hormones (FSH:
5.7 mU/mL (1.5-12.4); LH: <0.1 mU/mL (1.7-8.6); testosterone: <0.025 ng/mL (1.88-8.82); cortisol: 1.97 µg/dL (6.2-19.4);
GH: <0.05 ng/mL (0.06-5); IGF-1:14 ng/mL (67-225)). Furthermore, brain magnetic resonance revealed a solid necrotic
mass involving the pituitary root, compatible with pituitary metastasis. Two weeks later, patient referred polyuria,
nocturia and polydipsia, laboratory exam revealed decreased levels of urine osmolality (231 mOsm/kg (50-1200)) and
density (1001 g/L (1010-1030)); while sodium and blood osmolality levels were higher than normality 153 mEq/L
(135-145) and 298 mOsm/kg (275-295) respectively. Endocrinology department confirmed diabetes insipidus.
All of the outcomes and syndromes were compatible with panhipopituitarism. Antitumor treatment and hormonal
substitutive therapy were administered; afterwards the patient experienced a clinical improvement. Plasmatic
hormonal levels and tumour markers were periodically controlled remaining within normal values.
CONCLUSIONS
Panhipopituhitarism caused by a metastasis is a very strange condition (only 2% of tumours are able to cause pituitary
metastasis) especially if is due to NET. Also, this disease has a wide range of clinical presentations and may usually
remain undiagnosed. That is why it must be considered in oncologic patients with compatible symptomatology,
including the performance of plasmatic levels of pituitary hormones in clinical protocols. This case illustrates the
importance of Clinical Laboratory in early diagnosis of this disorder.
Endocrinology
P0978
EFFECT OF OBESITY ON THE RELATIONSHIP BETWEEN OXIDATIVE STRESS AND PON1 ACTIVITY WITH HDL SUBCLASSES
IN PCOS
I. Perovic Blagojevic 6, J. Vekic 4, D. Macut 2, S. Ignjatovic 5, M. Miljkovic-Trailovic 3, A. Zeljkovic 3, I. Bozic-Antic 1, J.

Bjekic-Macut 7, A. Mastilovic 6, J. Kotur-Stevuljevic 3
1
Clinic of Endocrinology, Diabetes and Diseases of Metabolism, Clinical Center Serbia, Belgrade, Serbia; Faculty of Medicine,
University of Belgrade, Belgrade, Serbia
2
Clinic of Endocrinology, Diabetes and Diseases of Metabolism, Clinical Center Serbia, Belgrade, Serbia; Faculty of Medicine,
University of Belgrade, Belgrade, Serbia;
3
Department for Medical Biochemistry, Faculty of Pharmacy, University of Belgrade, Belgrade, Serbia
4
Department for Medical Biochemistry, Faculty of Pharmacy, University of Belgrade, Belgrade, Serbia;
5
Department for Medical Biochemistry, Faculty of Pharmacy, University of Belgrade, Belgrade, Serbia; Center for Medical
Biochemistry, Clinical Center Serbia, Belgrade, Serbia;
6
Department of Laboratory Diagnostic, Clinical Hospital Center „Dr Dragisa Misovic-Dedinje“, Belgrade, Serbia
7
University Medical Center „Bezanijska kosa“, Belgrade, Serbia; Faculty of Medicine, University of Belgrade, Belgrade, Serbia
BACKGROUND-AIM
Background/Aim: Dyslipidemia is the most common metabolic abnormality in polycystic ovary syndrome (PCOS).
In obese PCOS women, altered lipid profile is commonly present, characterized by elevated concentrations of
small, dense low- density lipoprotein (sdLDL) particles, which are particularly atherogenic form of lipoproteins,
hypertriacylglycerolaemia and lowered high-density lipoprotein cholesterol (HDL-C) concentrations (LDL B phenotype).
High-density lipoprotein (HDL) constitutes a heterogeneous group of particles and the data on HDL particles
distribution and functionality in PCOS are scarce. Considering that paraoxonase 1 (PON1) is an antioxidative enzyme
located on HDL particles, the aim of this study was to analyse the connection between oxidative stress (OS) and PON1
activity with HDL subclasses in PCOS depending on obesity.
METHODS
Methods: Study included 115 women, aged 20 to 39 years, with proven PCOS (60 normal weight, BMI ≤ 25 kg/m²,
55 obese, BMI >25 kg/m²). Vertical polyacrylamide gradient gel electrophoresis was used to determine lipoprotein
subclasses distributions. OS status was assessed by total oxidative status (TOS), advanced oxidation protein
products (AOPP), malondialdehyde (MDA), prooxidant-antioxidant balance (PAB), total antioxidative status (TAS) and
superoxide dismutase (SOD) and PON1 activity.
RESULTS
Results: We found that obese patients had lower HDL size and higher proportion of HDL 3a subclasses (P<0.05). PAB
showed significant negative correlation with HDL 2a (r=-0.369; P<0.001), whereas MDA and SOD correlated positively
with HDL 3 subclasses (r=0.225; r=0.187; P<0.05). Serum PON1 activity was positively associated with proportions of
PON1 activity on HDL 2b (P<0.05) and 2a (P<0.01), but negatively with the proportion on HDL 3 particles (P<0.01).
Compared to LDL A, LDL B phenotype patients had increased TAS (P<0.05), SOD (P<0.05) and PON1 (P<0.001) activity
on HDL 2b, but decreased PON1 activity on HDL 3 subclasses (P<0.01).
CONCLUSIONS
Conclusion: Results of this study showed that OS is associated with atherogenic changes in lipoprotein subclasses
distribution in PCOS patients. Obesity in PCOS affects the profile of HDL subclasses, reflected through the reduced
proportion of PON1 activity on HDL 3 subclasses in the presence of sdLDL particles.
Endocrinology
P0979
A MIDDLE AGED MAN WITH CHANGING APPEARANCE - A CASE REPORT ON ACROMEGALY
T.U. Athapaththu 1, S. Peduruhewa 1

1
Department of Chemical Pathology, Colombo South Teaching Hospital, Kalubowila
BACKGROUND-AIM
Acromegaly is an endocrine disease caused by persistent hypersecretion of growth hormone (GH) and insulin-like
growth factor-1 (IGF-1) resulted in distinctive clinical presentation of acral enlargement. More than 95% of patients
with acromegaly have a GH-secreting pituitary adenoma arising from somatotroph cells of the pituitary gland and
resulted in hypersecretion of GH and IGF1. Less number of cases (<5% ) are due to excess GHRH-secreting hypothalamic
tumours or neuroendocrine tumours and rarely caused by ectopic GH-producing tumours. The elevated GH and IGF-I
levels in acromegaly lead to a number of cardiovascular, respiratory, endocrine, and metabolic comorbidities.
METHODS
A 52- year old previously healthy, male presented with 2 -year, history of achieving coarse facial features and,
progressive enlargement of hands & feet associated with numbness and recent history of intermittent headache and
visual impairment. On examination, he was overweight, blood pressure was elevated and B/L hemianopia was noted.
Initial work up revealed that he is having elevated IGF-1 levels and non-suppressible GH level following 75 g glucose
challenge test and diagnosed as acromegaly. Other pituitary hormones remained within normal levels. He had impaired
glucose tolerance, hypercalcaemia and hypertriglyceridemia. 2D ECHO revealed left ventricular hypertrophy. MRI
pituitary confirmed a microadenoma.An ectopic GH secreting tumour was excluded by performing CT abdomen and
chest. later, he underwent transsphenoidal resection of the pituitary tumour and he could achieve a better clinical
outcome.
RESULTS
Our patient had most of the clinical manifestations attributed to acromegaly along with features of mass effect.
The metabolic complications he had were, impaired glucose tolerance due to insulin antagonist effect of IGF 1,
hypertriglyceridemia due to lipolysis and hypercalcemia due to increased bone turnover by excess GH.
CONCLUSIONS
The gold standard tests to diagnose acromegaly are IGF-1 and glucose challenge test while the success of the surgery
can be evaluated by monitoring GH levels. Even though it is a rare disease, if left untreated, can lead to disfigurement
and significant morbidity and mortality. Thus, early diagnosis and treatment are essential in improving patient
outcome.
Endocrinology
P0980
CAUSES OF AZOOSPERMIA IN PATIENTS UNDERGOING TESTICULAR BIOPSY
M. Caparros Guerrero 1, A.M. Ceron Moreno 1, P.J. Sanchez Carretero 1, V. Martinez Pina 1, M. Arnaldos Carrillo 1, M.
Exposito Garcia 1, Y. Mestre Terkemani 1, M.I. Navarro Rey 1, R. Sansano Galiano 1, S. Requena Lopez 1, C. Guirao Blazquez
1
, M. Martinez Villanueva 1, I. Cebreiros Lopez 1, A.M. Martinez Lopez De Castro 1, A. Martinez-Escribano Garcia-Ripoll
1
, T. Casas Pina 1, J.A. Noguera Velasco 1
1
HOSPITAL CLÍNICO UNIVERSITARIO VIRGEN DE LA ARRIXACA
BACKGROUND-AIM
Testicular biopsy is currently the last reproductive option for men with azoospermia. It consists of the surgical
extraction of a small section of testicular tissue in order to recover sperm from it that can be cryopreserved and used
in an assisted reproduction process. Few public hospitals currently perform this technique.
METHODS
We collected biopsy samples from the left and right testicles of 70 men with azoospermia and infertility of at least
1 year of evolution. The period of study was since May 2019 to January 2023. The samples were analyzed in a phase
contrast microscope to evaluate the presence of spermatozoa, reporting the result as: absence, scarce (1-2 per field),
moderate (2-10 per field) and abundant (>10 per field). In the cases in which spermatozoa were found, the tissue
sample was cryopreserved, for which Origio's Sperm Freezing Medium was used.
RESULTS
The age range of the men who underwent biopsy was 13-47 years. Spermatozoa were found in 47.1% of patients,
so their samples were cryopreserved. Three patients (4.3%) were diagnosed with cystic fibrosis, which suggests an
obstructive azoospermia. Five patients (7.1%) were diagnosed with Klinefelter syndrome: no spermatozoa were found
in any of them, but the samples were cryopreserved at the clinician's request. Factors possibly related to azoospermia
included decreased testicular size (14.3%), varicocele (11.4%), hypogonadotrophic hypogonadism (7.1%), removal of
one of the testicles (2.9%) or endocrine disorders such as obesity (8.6%), diabetes (5.7%) or hypothyroidism (4.3%).
A 15.7% reported toxic habits (smoking). In 41 of the men, at least one hormonal alteration was found. The most
frequent were: FSH elevation (50%), LH elevation (24.3%), prolactin elevation (20%) and decreased testosterone (10%).
No association was observed between the number of spermatozoa found and the pathology of the male. In a 11.4% of
the patients, no spermatozoa were found in the biopsied tissue despite having a normal hormonal profile.
CONCLUSIONS
Testicular biopsy is an interesting reproductive option for those men in whom there is no option of obtaining a viable
sperm sample. The most frequent alterations found in these patients are decreased testicular size and elevated FSH.
Endocrinology
P0981
CUSHING'S DISEASE WITHOUT CONCLUSIVE RADIOLOGICAL IMAGES. CONTRIBUTION OF INFERIOR PETROSAL SINUS
CATHETERISATION TO THE DEFINITIVE DIAGNOSIS.
D. Casanova Expósito 1, C. Valldecabres Ortiz 1, S. Górriz Pintado 1

1
Biological Diagnostics Area, Hospital Universitario de la Ribera
BACKGROUND-AIM
Cushing's syndrome is characterised by signs and symptoms resulting from an excessive increase in blood cortisol
concentration.
In the case of adrenocorticotropic hormone (ACTH)-dependent Cushing's syndrome, it is called Cushing's disease, and
is usually caused by the presence of a pituitary microadenoma.
When imaging techniques such as magnetic resonance imaging (MRI) are not sufficient for the diagnosis of an ACTH-
producing microadenoma, bilateral inferior petrosal sinus catheterisation may be used for differential diagnosis
between pituitary tumour and ACTH-secreting ectopic tumour. Positive test criteria for pituitary Cushing's is given by
a central-peripheral gradient equal to or greater than two in baseline samples or equal to or greater than three in the
maximum obtained after stimulation with corticotropin-releasing hormone (CRH).
METHODS
47-year-old morbidly obese woman. Following treatment failure with dietary measures alone, possible secondary
causes of obesity were investigated. Complementary tests revealed findings suggestive of endogenous
hypercortisolism (elevated ACTH, salivary cortisol and urinary free cortisol, above the reference limit) associated with
hypersecretion of insulin-like growth factor 1 (IGF1).
MRI with and without contrast was performed, in which no alterations in the pituitary gland were observed.
Following these findings, it was decided to perform bilateral catheterisation of the inferior petrosal sinuses.
RESULTS
A bilateral femoral venous puncture was performed and a right 6F and a left 5F introducers were placed. Using vertebral
catheters and guidewire, selective catheterisation of both petrosal sinuses was realized. Stimulation with 100 µg CRH
was performed and blood was simultaneously withdrawn from both petrosal sinuses and the peripheral vein at 5, basal,
1, 3, 5, 15 and 30 min after CRH administration.
The results showed that ACTH levels collected in the left sinus had higher values. A left/peripheral ratio greater than
3 was obtained at 3 and 5 minutes, indicative of lateralisation.
CONCLUSIONS
Although analytical data suggest hypercortisolism, imaging tests do not show the responsible pituitary microadenoma.
In this case, bilateral inferior petrosal sinus catheterisation is a very useful test to identify the left lateralisation of
the microadenoma.
Endocrinology
P0982
HYPOPITUITARISM AS A SEQUALAE OF EMPTY SELLA SYNDROME – A DELAYED PRESENTATION OF SHEEHAN’S
SYNDROME
T.M.N.K. Puliyadda 1, G. Premawansa 2, B.K.T.P. Dayanath 1

1
Department of Chemical Pathology, Colombo North Teaching Hospital , Sri Lanka
2
Department of Medicine, Colombo North Teaching Hospital, Sri Lanka
BACKGROUND-AIM
The herniation of sub-arachnoid space into sella causing flattening of pituitary gland is defined as empty sella
syndrome (ESS). Sheehan’s syndrome (SS), the necrosis of pituitary gland due to infarction or ischaemia following
postpartum hemorrhage (PPH) is a known cause for ESS. It can present as hypopituitarism either acutely or in a delayed
manner.
METHODS
Case Presentation
A 45-year-old female presented with myalgia, lethargy and dyspeptic symptoms for 6 months duration. She had
developed secondary amenorrhea, lactation failure and loss of pubic/axillary hair one year following her uncomplicated
second childbirth at the age of 27 and the clinic follow-up was defaulted. On general examination, she was pale but
there was no ecchymotic patches or bruises. A loss of axillary and public hair was noted with bilateral involution of
breasts.
RESULTS
The investigations showed hyponatremia and anemia. A low normal thyroid-stimulating hormone (TSH) value of 1.0
mIU/L (0.465 – 4.68) with a low free T4 value of 2.3 pmol/L (10 – 28.2) was indicative of secondary hypothyroidism.
Further evaluation revealed a low growth hormone level of < 0.08 µg/L (2 – 5) and insulin like growth factor level of
48 ng/mL (124 – 290). A low cortisol [10 nmol/L (138 – 635)] and adrenocorticotropic hormone level [8 pg/mL (7 –
41)] were identified. Her serum gonadotropin levels were well below the reference range and estradiol level was in
post-menopausal range. Hence, the diagnosis of hypopituitarism was established. The MRI scan of brain confirmed the
diagnosis of empty sella. According to her classic clinical presentation, ESS was probably secondary to SS. A polyuria
was noted after initiation of steroids and could be due to presence of pre-existing partial central diabetes insipidus
masked by hypocortisolism. The symptoms got improved following hormone replacement therapy.
CONCLUSIONS
SS can also occur due to extremely low blood pressure or vascular spasms during childbirth even without a history of
PPH. However, it is important to exclude mimics of SS like hypophysitis and adenoma due to the delayed presentation.
This also emphasizes the importance of performing both TSH and free T4 specifically when suspecting secondary
hypothyroidism.
Endocrinology
P0983
OCCULT INSULINOMA – THE IMPORTANCE OF SELECTIVE INTRA-ARTERIAL CALCIUM STIMULATION TEST IN
PREOPERATIVE LOCALIZATION
T.M.N.K. Puliyadda 1, B.K.T.P. Dayanath 1

1
Department of Chemical Pathology, Colombo North Teaching Hospital , Sri Lanka
BACKGROUND-AIM
Insulinoma is a functional neuroendocrine tumour of pancreas which causes hyper-secretion of insulin. It is the
commonest cause of hypoglycaemia in a healthy individual when there is no evidence of factitious hypoglycaemia.
Occult insulinoma is a biochemically proven tumour with inability to determine the anatomical site prior to surgery.
Selective intra-arterial calcium stimulation test (SIACS) has ≥ 94% sensitivity in preoperative localization of an occult
insulinoma.
METHODS
Case Presentation
A 46-year-old male presented with frequent episodes of faintishness for 7 months duration. It was associated with
autonomic symptoms and blurring of vision with no confusion or seizures. It got worsen following exercise and there
were no episodes occurring in postprandial period. The symptoms disappeared following ingestion of food.
RESULTS
The investigations revealed a fasting plasma glucose level of 63 mg/dL (74–100). The 72-hour prolonged fasting test
which is the gold standard in biochemical diagnosis of endogenous hyperinsulinaemia indicated a positive result with
an increased serum insulin level of 185.01 pmol/L (<18) and C-peptide level of 0.86 nmol/L (<0.2) when having adequate
hypoglycaemia (53 mg/dL). As non-invasive radiological findings were negative, an endoscopy guided ultrasound scan
of pancreas was performed. It revealed a focal lesion (0.9 x 0.9 cm) in head of the pancreas which was inconsistent with
typical features of an insulinoma. Therefore, it was followed by a SIACS test and it showed a two-fold rise in insulin
level from the baseline following stimulation of gastro duodenal artery. It confirmed the presence of an insulinoma
in the head of pancreas. As he refused undergoing surgical intervention, he has been being followed up with blood
glucose monitoring and frequent diet therapy.
CONCLUSIONS
In SIACS test, the intra-arterially given calcium stimulates secretion of insulin only from the abnormally hyper-
functioning beta cells of pancreas. Hence, it is useful in preoperative localization of an occult insulinoma, identifying
multiple lesions and further confirmation of identified focal lesions. This case also emphasizes the importance of
considering insulinoma as a differential diagnosis in a healthy individual with symptoms of hypoglycaemia.
Endocrinology
P0984
SET UP OF A ISOTOPIC DILUTION LC-MS/MS METHOD FOR STEROID QUANTIFICATION IN SEMINAL FLUID AND ITS
APPLICATION IN A COHORT OF INFERTILE MEN
S. Marchiani 2, F. Villanelli 2, S. Dabizzi 1, S. Degl'Innocenti 1, M.G. Fino 1, L. Vignozzi 2, G. Danza 2

1
Andrology, Female Endocrinology and Gender Incongruence Unit, Careggi University Hospital, Florence, Italy
2
Department of Experimental and Clinical Biomedical Sciences Mario Serio, University of Florence, Florence, Italy
BACKGROUND-AIM
Nowadays couple infertility affects about 15% of couples and the male factor is implicated in about 40% of cases.
Semen analysis is the corner stone for male infertility diagnosis; however, the biochemical composition of the
seminal fluid (SF) is poorly investigated, and steroid hormones are scarcely considered despite their essential role in
spermatogenesis and function of the sexual accessory organs. Steroid measurement in SF could help to understand
their potential utility as biomarkers for male infertility.
The aim of this study was to develop an isotopic dilution LC-MS/MS method for steroid quantification, both in serum
and SF, to compare their levels in a cohort of infertile men.
METHODS
We determined the levels of Androstenedione (A), Testosterone (T), Dheidroepiandrosterone (DHEA),
Dheidroepiandrosterone sulphate (DHEAS), Dihydrotestosterone (DHT), Progesterone (P), 17-Hydroxy-Progesterone
(17-OH-P), Cortisol (F) and Cortisone (E) in serum and SF samples of 42 infertile men. 50 µL of serum or SF were added
to the internal standard mix in acetonitrile, centrifuged, diluted with water and injected in the system (1260 Agilent
HPLC coupled with a 6500 QTRAP mass spectrometer). We used a C18 5 µm 20×2 mm as preconcentrating column and
Luna C18 3 µm 50x2 mm as analytical column. Quantitation was done using a seven-points curve (Chromsystems) and
the MULTIQUANT software (Sciex).
RESULTS
The levels of A, T, DHT, DHEA, DHEAS, F and E were significantly lower in SF compared to serum. 17-OH-P was not
detectable in SF and P was detected only in 5 subjects. Interestingly, a significant positive correlation was found
between serum and SF levels for A, DHEA, DHEAS but not for T, DHT, F and E indicating that SF reflects a “local” difference
for these hormones that deserves great attention considering their role in fertilization process.
CONCLUSIONS
We developed a simple and reliable method for steroids quantification in both SF and serum that could be successfully
used in routine clinical practice after its validation. Our preliminary results suggest a possible role of some steroid
hormones evaluated in SF as novel biomarkers for male infertility.
Endocrinology
P0985
PATTERN OF THYROID FUNCTION ABNORMALITIES IN PATIENTS WITH DEPRESSION IN A TERTIARY HOSPITAL,
SOUTHWESTERN NIGERIA
M. Bello 1, A. Ajose 1, K. Mosaku 2, T. Adedeji 1, A. Ajeigbe 1

1
Department of Chemical Pathology, Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, Osun State, Nigeria.
2
Department of Mental Health, Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, Osun State, Nigeria.
BACKGROUND-AIM
BACKGROUND: Depression is one of the leading causes of disability globally, with complex and multifactorial etiology.
Biochemical abnormalities like thyroid dysfunction have been reported in depression albeit with conflicting results.
AIM: To examine the prevalence and pattern of thyroid function abnormalities in patients with depression.
METHODS
MATERIALS AND METHODS: In this observational case-control study, 60 participants with depression were recruited
from The Mental Health Department of Obafemi Awolowo University Teaching Hospitals Complex (OAUTHC), Ile-
Ife, while controls were 60 age and sex-matched apparently healthy hospital staff. A 17-item Hamilton Depression
Rating Scale (HAMD17) was used to evaluate depression. Then, 5 milliliters of venous whole blood was collected
aseptically and serum harvested for assay of thyroid hormones using electrochemiluminescence immunoassay (ECLIA)
on the cobas E 411 analyser (Roche Diagnostics GmbH, SandhoferStrasse 116, D-68305 Mannheim, Germany) at The
Department of Chemical Pathology (OAUTHC, Ile-Ife). Results of the thyroid function were evaluated to give a clinical
diagnosis. Analysis was done using descriptive statistics in the Statistical Product and Service Solutions (SPSS 25 IBM,
USA), and presented as frequencies in table, pie charts and histogram.
RESULTS
RESULTS: The participants with depression (36.18±12.7 years) were slightly older than the controls (34.47±7.8 years).
The prevalence of thyroid function abnormalities in the study was 21.7% (13 patients) while 78.3% (47 patients)
was euthyroid. The pattern of thyroid function abnormalities observed were subclinical hypothyroidism followed by
sick euthyroid syndrome and secondary hypothyroidism representing 46.1%, 38.5% and 15.4 % of abnormal thyroid
function status respectively.
CONCLUSIONS
CONCLUSION: This study found that thyroid function abnormalities are not uncommon among participants with
depression and that the prevalence is as high as 21.7%. Subclinical hypothyroidism appears to be the most prevalent
thyroid function abnormality among participants with depression in this environment. Hence, routine screening to
assess thyroid function status among depressed patients could be an adjunct in the management of depression.
Endocrinology
P0986
VITAMIN D TOXICOSIS IN A PATIENT WITH RENAL IMPAIRMENT. CASE REPORT
B. Luis Sánchez 1, A. Caro Pradillos 1, N. Rico Ríos 1, M. Calero Ruiz 1, A. Sáez-Benito Godino 1
1
UGC Laboratorio. Hospital Universitario Puerta del Mar, Cádiz (Spain).
BACKGROUND-AIM
Vitamin D (VD) deficiency is relatively common (particularly in low sunlight exposure areas and risk populations) and
supplementation is standard practice. It may entail some risks, including hypercalcemia and its associated symptoms.
Due to VD toxicity risk and the essential role of the laboratory in its diagnosis and management, we introduce this
case report.
METHODS
A 77-year-old patient with chronic kidney disease (CKD) presented with poor general condition, low level of
consciousness and neurological deterioration to the emergency department. Post-admission bloods highlighted a
critical calcium (Ca) result (3,5 mmmol/L) in conjunction with raised creatinine and urea. The clinical laboratory
contributed to the diagnosis work-up with reflecting testing.
RESULTS
The renal profile was consistent with underlying CKD. The laboratory alerted the critical Ca to the clinical team
for immediate intervention and discussed the results of tests reflected: Serum protein electrophoresis=normal –
Myeloma (malignancy) ruled out; Parathormone=9pg/mL (22-101)– Hyperparathyroidism ruled out; VD=295nmol/L–
Intoxication identified as the cause of critical hypercalcemia.
According to medical records, the patient was on VD (266 ucg/fortnight), but the posology was wrongly interpreted
and VD was taken daily for months leading to an accumulation (VD long half-life) and causing toxicity. VD excess
increased the absorption of Ca triggering a hypercalcemia with severe symptoms associated such as the low level
of consciousness. Patient also suffered de novo atrial fibrillation and underwent haemodialysis. VD treatment was
immediately wihtdrawn. Ca level gradually normalised and patients clinical situation improved.
CONCLUSIONS
The communication of critical results and the utility of reflecting testing is essential for a timely and appropriate
patient management. Although VD intoxications are rare, there are situations (prescription/interpretation errors,
accidental ingestion, absorption problems,etc.) where its probability increases. For this reason, once treatment has
been established, VD concentration should be monitored and laboratory should identify emerging hypercalcemia in
patients with supplementation, specially in risk population.
Endocrinology
P0987
ARE METHOD SPECIFIC CUTOFFS FOR SERUM GLUCOSE, 25-HYDROXYVITAMIN D AND CORTISOL RESPONSE TO
ADRENOCORTICOTROPHIN TEST NEEDED?
F.F.M. Chay 1, H.L. Yeo 1, A.X.W. Ang 1, Q.T.B. Goh 1, S.Q. Lee 1, A. Omar 1, K. Lim 1, L. Ong 1
1
Alexandra Hospital
BACKGROUND-AIM
Endocrine testing such as glucose (fasting or 2 hours after glucose tolerance testing), 25-hydroxyvitamin D (25OHD)
and cortisol response after adrenocorticotrophin injection, are interpreted using fixed international decision limits
that are not method specific. The impact of analytical bias on diagnostic interpretation is uncertain and varied across
published studies.
We compared the method comparison between Abbott Alinity and Beckman and reviewed the impact of analytical bias
on diagnosis of diabetes mellitus, vitamin D deficiency and adrenal insufficiency.
METHODS
Using method comparison between Abbott Alinity ci and Beckman DxC (for glucose) and Beckman DXI (for 25OHD and
cortisol), we reviewed the correlation at clinical decision limits for diagnosis of diabetes mellitus (using glucose above
6.1 mmol/L), vitamin D insufficiency and deficiency (25OHD below 30 ug/L) and adrenal insufficiency (using cortisol
post adrenocorticotrophin target of 500 nmol/L).
Significant bias was considered if the overall or clinical decision limit cutoffs differences exceeded the Royal College
of Pathologists of Australasia (RCPA) analytical performance specifications (APS) published in 2019.
RESULTS
Method comparison between Alinity ci and Beckman DxC for glucose (N=40) was comparable overall and at clinical
decision limits. 32% of samples were reassigned as non-diabetic on Alinity compared to Beckman.
Method comparison for 25OHD (N=40) was comparable overall and at clinical decision limits. 33% of samples previously
defined as sufficient vitamin D were reassigned as insufficient vitamin D on Alinity. No samples were newly defined
as vitamin D deficient.
Method comparison for cortisol (N=40) was comparable overall and at clinical decision limits. 38% of samples
previously defined as adrenal sufficiency were reassigned as adrenal insufficient on Alinity. All samples that were
defined as adrenal insufficient on Beckman and Alinity concurred.
CONCLUSIONS
Using RCPA APS, there was good method comparison overall and at clinical decision limits for glucose, 25OHD and
cortisol, with non-significant bias. There was 32-38% of samples that were reclassified on Alinity, which highlighted
potential impact to clinical management.
Endocrinology
P0988
HYPERANDROGENISM IN POLYCYSTIC OVARY SYNDROME
E. Talbi 1, T. Mazouni 1, R. Ghodbane 1, L. Zerelli 1, S. Oueslati 1, S. Hammami 1, R. Mahjoub 1

1
Clinical Biology Laboratory, Zouhair Kallel Institute of Nutrition and food technology, Tunis
BACKGROUND-AIM
Obesity induces abnormalities in sex steroids balance, particularly androgens. Hyperandrogenism is also the founding
element of polycystic ovary syndrome (PCOS) and a cause of infertility. The aim of the study was to evaluate biological
hyperandrogenism in morbidly obese women in relation to PCOS.
METHODS
This cross-sectional study included 50 women with morbid obesity (BMI ≥ 40 Kg/m²). Patients on estrogen-
progestin therapy and those with elevated TSH, prolactin or 17-hydroxy progesterone were excluded. The diagnosis
of PCOS was retained according to the Rotterdam 2003 criteria. Total testosteronemia (TT) was determined by
immunoChemiluminescence and delta-4-androstenedione (∆-4A) and dehydroepiandrosterone sulfate (DHEAS) by
radioimmunological methods.
RESULTS
Total testosteronemia levels (0.32±0.14 vs. 0.223±0.13 ng/mL; p=0.07) were higher in PCOS + women. In contrast, ∆-4A
(2.8±1.0 vs. 2.4±0.9 ng/mL) and DHEAS (1338±772 vs. 1457±917 ng/mL) levels did not differ between the two groups.
Hyper testosteronemia (0.6 ng/mL) was observed in 10% of SPOK+ women versus 0% of SPOK - women.
Elevated ∆-4A (>3 ng/mL) was found in 25% of PCOS + and 20% of PCOS - women.
No women in either group had elevated DHEAS levels (>3850 ng/mL).
CONCLUSIONS
Total testosterone assay remains the first-line test to explore biological hyper androgenism in obese women and may
help in the diagnosis of PCOS.
Endocrinology
P0989
LABORATORY DIAGNOSIS OF INSULINOMA,CASE RAPORT
R. Arapi 1, B. Cullhaj 2
1
Department of Laboratory Medicine, University of Medicine, University Hospital Center "Mother Teresa"
2
GeniusLAB-Labioratory Network
BACKGROUND-AIM
Insulinoma is a insulin secreting tumor. It is a rare tumor and the diagnose can be difficult and it can take a long time.
Insulinoma is usually located inside the pancreas but ectopic cases have been reported.
METHODS
Tests that determine the diagnose are high c-peptide, high insulin and high proinsulin following episodes of
hypoglycemia. C-peptide and insulin were performed with chemiluminescence immunoassay (CLIA) and enhanced
chemiluminescence immunoassay (ECLIA) respectively , prolinsulin was performed with enzyme immunoassay (EIA).
RESULTS
Case report: A 48 year old female patient presents to the emergency room with tremor, sweating and mental fog. This
was her 4 visit to the ER in the last 2 month. The onset of symptoms was 1 year prior with tiredness and irritability
during the day .Her symptoms were treated as anxiety attacks in the past and laboratory tests were not performed .The
blood tests showed a glycemia of 35 mg/dl (70-110 mg/dl). She was admits for further evaluation. The laboratory
tests showed a c-peptid of 8.176 ng/ml (0.3-3.73 ng/ml) , insulin level of 35.66 uIU/ml (2.6-24.9 uIU/ml) and later
a proinsulin of 113 pmol/l (<11 pmol/l). After the test results came an abdominal computed tomography (CT) was
performed showing a singular mass in the tail of the pancreas .After the surgical removal of the mass blood glucose
levels reached normal levels without further hypoglycemic episodes.
CONCLUSIONS
Insulinomas in the beginning can be misdiagnosed or neglected so they can be difficult to diagnose and it needs
repeated evaluation and good communication between medical specialties and the patient.
Endocrinology
P0990
A NOVEL METHOD FOR THE FREE THYROXINE (FT4) CONCENTRATION MONITORING IN A DRIED BLOOD SPOT (DBS)
SAMPLES.
A. Krol 2, K. Bornikowska 1, M. Ostrowska 1, P. Glinicki 1, W. Zgliczynski 1, K. Kowalski 2

1
Department of Medical Statistics, School of Public Health, Centre of Postgraduate Medical Education, 01-826 Warsaw,
Poland
2
Masdiag Sp. z o. o., 01-882 Warsaw, Poland
BACKGROUND-AIM
FT4 is the non-protein-bound plasma fraction of thyroxine, the main hormone produced by the thyroid gland.
Simultaneous determination of FT4 and thyroid stimulating hormone (TSH) concentrations is fundamental for the
diagnosis of thyroid functional status. In 2019 the average number of FT4 diagnostic tests performed in Poland reached
over 710 000 samples. In light of the increasing number of tests performed each year, allowing patients to collect
samples at home and send them to a laboratory could revolutionize the diagnostic process due to easy collection and
high sample stability. The aim of the study was to develop a DBS–based method for FT4 determination.
METHODS
The developed method is based on solid-phase antigen linked technique (SPALT) and chemiluminescent detection
performed on a DiaSorin LIAISON XL autoanalyzer. For preliminary comparisons of the methods, serum and DBS
samples were collected simultaneously from each of 59 patients with (n=12) and without (n=47) thyroid disorders.
Serum samples were analyzed directly, while DBS samples had to be extracted beforehand. From each DBS sample
two ø4.6 mm discs (~12 µl blood) were punched into a 96 well plate and incubated (1000 rpm, 37oC) with 250 µl of
extraction buffer.
RESULTS
Analytic sensitivity of the test given in the assay protocol was 1,28 pmol/l. An updated reference range of FT4 in
DBS was calculated according to CLSI EP28-A3C guideline, based on strong logarithmic correlation between DBS and
serum FT4 results (7,51–12,41 pmol/l). According to the results obtained from both tested materials, this method had
a sensitivity value of 84,62% and specificity of 97,83%.
CONCLUSIONS
The developed method could serve as a diagnostic tool for thyroid function monitoring in the general population.
Furthermore, its high sensitivity and specificity could make it a suitable component of newborn screening programs
for the detection of congenital hypothyroidism together with the measurement of TSH concentrations in DBS samples.
Endocrinology
P0991
A NOVEL, SENSITIVE DRIED BLOOD SPOT (DBS) BASED METHOD FOR THYROID STIMULATING HORMONE (TSH)
MONITORING IN THE GENERAL POPULATION.
A. Krol 2, K. Bornikowska 1, M. Ostrowska 1, P. Glinicki 1, W. Zgliczynski 1, K. Kowalski 2

1
Department of Medical Statistics, School of Public Health, Centre of Postgraduate Medical Education, 01-826 Warsaw,
Poland
2
Masdiag Sp. z o. o., 01-882 Warsaw, Poland
BACKGROUND-AIM
Thyroid hormones are homeostasis-maintaining hormones that target almost every tissue in the body. Diseases such
as hypothyroidism and hyperthyroidism affect all populations worldwide. In Poland, the number of TSH diagnostic
tests in 2019 reached over 1,75 mln samples and is growing every year. Thanks to easy at-home sample collection and
shipment, the use of DBS for thyroid function monitoring could revolutionize the diagnostic process. The purpose of
this study was to develop and validate a DBS-based method for sensitive TSH determination in the general population.
METHODS
The method developed is based on the sandwich immunochemiluminescence technique performed on the DiaSorin
LIAISON XL autoanalyzer. For preliminary method comparison, serum and DBS samples were collected simultaneously
from each of 246 patients with hyperthyroidism (n=30), hypothyroidism (n=31) and without thyroid disorders (n=185).
Serum samples were analyzed directly, while DBS samples had to be extracted beforehand. From each DBS sample
two ø4.6 mm discs (~12 µl blood) were punched into a 96 well plate and incubated (1000 rpm, 37oC) with 350 µl of
extraction buffer.
RESULTS
An updated reference range of TSH in DBS was calculated according to CLSI EP28-A3C guideline, based on strong linear
correlation between DBS and serum TSH results (0,018–0,084 mIU/ml). Analytic sensitivity of the TSH test given in the
assay protocol was 0,004 mIU/ml. This DBS-based method sensitivity value was 83,3% for hyperthyroidism and 87,1%
for hypothyroidism, while specificity was 98,3% and 98,9%.
CONCLUSIONS
In conclusion, the developed method has demonstrated its usefulness as a tool for monitoring thyroid gland function
in the general population. Additionally, the analytical sensitivity of this method is significantly higher than that of
newborn screening tests and point-of-care tests available commercially.
Endocrinology
P0992
IMPACT OF THE TEST METHODOLOGY ON THE PREVALENCE OF SUBLINICAL HYPOTHYROIDISM AND LOW FT4 LEVELS
WITH NORMAL TSH IN OUR HEALTH CARE AREA.
M.C. Martín Fernández De Basoa 1, M.T. Concepción Masip 1

1
Hormone, Tumor Markers and Prenatal Screening Unit of the Clinical Analysis Department. Complejo Hospitalario
Universitario Nuestra Señora de Candelaria, Santa Cruz de Tenerife
BACKGROUND-AIM
Subclinical hypothyroidism is considered a disorder that occurs in generally asymptomatic individuals, and is
characterized by the finding of elevated levels of thyroid-stimulating hormone (TSH) with normal levels of thyroid
hormones (FT4 and FT3). Despite being a frequent problem, its therapeutic management and its clinical significance
are controversial. It has been estimated according to different studies that its prevalence in the Spanish population
is 7.9%.
The aim of the present study is to analyze whether the prevalence of subclinical hypothyroidism and discordant FT4
values with normal TSH in our healthcare area varies according to the technology used, since with the one currently
used in our laboratory (Roche Diagnostics electrochemiluminescence) the prevalence of subclinical hypothyroidism is
very high. higher than that reported in the literature (11.7%).
METHODS
863 TSH samples and 322 FT4 samples were analyzed by two immunoassays:
1) Electrochemiluminescence from Roche Diagnostics (cobas e801).
2) Chemiluminescence with acridinium esters from Abbott Científica, S.A (Alinity i).
RESULTS
With Roche Diagnostics the prevalence of subclinical hypothyroidism was 11.7% (95 cases) and the prevalence of
discordant FT4 with normal TSH was 7.76% (25 cases).
With Abbott Científica, S.A. the prevalence of subclinical hypothyroidism was 0,58% (5 cases) and the prevalence of
discordant FT4 with normal TSH was 0%.
CONCLUSIONS
Both prevalences show significant variability depending on the methodology used in the analysis.
In the case of discordant FT4 values with normal TSH, we were able to normalize all the samples by using Alinity i
(Abbott).
By changing the commercial house, we not only managed to improve the prevalence data, but we also achieved very
important economic savings, since in 95% of the samples analyzed it would not be necessary to perform the FT4 in the
case of falsely elevated TSH values. In addition, by not presenting discordant FT4 values, possible therapeutic errors
with the consequent iatrogenesis that would occur would be avoided.
Endocrinology
P0993
A NOVEL MODEL FOR ESTIMATING WITHIN-SUBJECT BIOLOGICAL VARIATION USING ROUTINE LABORATORY DATA
WITH SKEWED DISTRIBUTIONS
E.Å. Røys 3, K. Viste 4, J.D. Graham 2, N.A. Guldhaug 4, B. Alaour 6, M.S. Sylte 3, J. Torsvik 3, H. Strand 5, M. Marber 6, T.
Omland 5, E. Theodorsson 1, K.M. Aakre 4
1
Department of Biomedical and Clinical Sciences, Division of Clinical Chemistry and Pharmacology, Linkoping University,
Sweden.
2
Department of Chemical Pathology, SydPath, St. Vincent's Hospital, Sydney, Darlinghurst, NSW, Australia.
3
Department of Medical Biochemistry and Pharmacology, Haukeland University Hospital, Bergen, Norway.
4
Hormone Laboratory, Department of Medical Biochemistry and Pharmacology, Haukeland University Hospital, Bergen,
Norway.
5
Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
6
King’s BHF Centre of Research Excellence, School of Cardiovascular Medicine and Sciences, King’s College London, United
Kingdom.
BACKGROUND-AIM
Biological variation data are useful for suggesting analytical quality specifications and clinical interpretation of
laboratory results. This study aims to develop a novel method for estimating within-subject biological variation (CVI)
from routine laboratory data with skewed distributions.
METHODS
Aldosterone was extracted from the laboratory data system at Haukeland University Hospital over a 3-year period.
Patients with at least two results were included and stratified by sex and presumed menopausal status: premenopausal
≤41 years, postmenopausal ≥55 years. Only samples taken between 8:00 AM and 10:00 AM were included, and the first
concentration was required to be within the local reference range. The between-result ratio was calculated. The µ, σ,
and λ from a BoxCox transformed ratio distribution fitted by RefineR were used to estimate the CVI with a lognormal
distribution. The BoxCox σ was reduced by √2, and by analytical variation (CVA). The CVI was finally estimated by
re-transforming the BoxCox to a value on the original scale. The approach was validated using Monte Carlo (MC)
simulations, repeated 5000 times, each with random ratio distributions formed by two mixture distributions: mixing a
Gaussian (CVA 1-10%) and lognormal (Geometric CVI: 20-60%). The results were compared to experimental data from
30 participants (16 women) with weekly samplings over ten weeks that were analyzed following the recommendations
of EFLM (CV-ANOVA).
RESULTS
The simulation showed minimal differences in geometric CVI between the randomly generated distributions and the
model results. The geometric CVI results for Aldosterone were 47% for men (1007 ratios), 48% for postmenopausal
(552 ratios), and 67% for premenopausal women (1632 ratios). In comparison, the experimental geometric CVI results
(CV-ANOVA) were: 49% (CI 43-58%) for men, 40% (CI 35-48%) for postmenopausal women, and 52% (CI 45-65%) for
premenopausal women.
CONCLUSIONS
CVIs may be estimated using routine laboratory data with a skewed distribution using a novel approach, including
BoxCox transformation and MC simulations. Data were similar to experimental data analyzed using the gold standard
EFLM method (2 of 3 subgroups). Care should be taken in cases of very high CVI estimates.
Endocrinology
P0994
ADDISONIAN CRISIS: CASE REPORT
C. Tapia Córdoba 1, P. Mayor Zapatero 1, M.P. Ocón Sánchez 1, J.F. Ruiz Escalera 1, M.M. Corzo Corbera 1
1
Hospital Regional Universitario de Málaga, Málaga
BACKGROUND-AIM
Primary adrenal insufficiency (PAI), also known as Addison's disease, is a rare endocrine disorder caused by the
destruction of the adrenal gland cortex. This generates a corticosteroid deficit which is manifested in a slow and non-
specific manner. However, it can also mannifest as an acute emergency in physiological stress situations due to this
lack of corticosteroids.
METHODS
A 4-years-old boy presented loss of reactivity of an hour of evolution, generalized rigidity, trismus, gaze deviated, fever
up to 38.5ºC in the last 2 days and catarrhal symptoms without vomiting. He has no known allergies to medications and
an adequate vaccination schedule for his age. On clinical examination he was in poor general condition, well hydrated
and perfused, normal cardiac auscultation and slight skin hyperpigmentation.
In view of hemodynamic instability and the onset of status convulsus, admission to the pediatric intensive care unit
was decided and antibiotic therapy was started on suspicion of septic shock.
RESULTS
Laboratory evaluation: glucose 25 mg/dL (normal range: 70–110), c- reactiv protein 147 mg/L (normal range < 5),
procalcitonin 24,68 ng/mL (normal range < 0,5), leukocytosis with neutrophilia and and metabolic acidosis with lactic
acid 2,5 mmol/L (normal range < 2).
In analytical control, basal cortisol was found to be decreased, so a Synacthen test was performed for suspected PAI.
It showed no cortisol response at 30 and 60 minutes after stimulation with synthetic adrenocorticotrophic hormone
(ACTH).
Treatment with hydrocortisone 12 mg/m²/day was started and progressive clinical improvement was observed in the
patient, who after several days was transferred to the ward and subsequently discharged with substitutive treatment
CONCLUSIONS
Adrenal crisis is a consequence of cortisol deficiency in a physiological stress situation, in which the demand of
corticosteroids increses while its production is insufficient or null. In this case report the main cause of the crisis is a
septic shock due to a respiratory infection, which revealed primary adrenal insufficiency. Laboratory tests, especially
the ACTH 1-24 stimulation test and the interpretation of the results obtained, are fundamental for the diagnosis and
establishment of treatment with hormone replacement as soon as posible.
Endocrinology
P0995
BLOOD LEUKOCYTE ROS PRODUCTION REFLECTS SEMINAL FLUID OXIDATIVE STRESS AND SPERMATOZOA
DYSFUNCTION IN IDIOPATHIC INFERTILE MEN
S. Borghi 3, F.R. Argento 3, E. Fini 3, A. Mannucci 3, R. Fucci 1, C. Giachini 1, R. Picone 1, G. Cito 1, N. Taddei 3, A. Fanelli
2
, T. Biagioli 2, M. Brogi 2, M.E. Coccia 4, M. Becatti 3, C. Fiorillo 3
1
Assisted Reproductive Technology Centre, Careggi University Hospital, Firenze, Italy.
2
Biochemistry Laboratory, Diagnostic Department, Azienda Ospedaliero Universitaria Careggi, Firenze, Italy.
3
Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Firenze, Italy
4
Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Firenze, Italy and Assisted
Reproductive Technology Centre, Careggi University Hospital, Firenze, Italy.
BACKGROUND-AIM
A large proportion of infertile males does not receive a clear diagnosis, being considered as idiopathic/unexplained
cases due to infertility diagnosis based on standard semen parameters. Particularly in unexplained cases, the search for
new indicators seems mandatory to provide specific information. In the etiopathogenesis of male infertility oxidative
stress displays important roles by negatively affecting sperm quality and function.
METHODS
In this study, performed in a population of 34 idiopathic infertile men and in 52 age-matched controls, we assessed
routine semen parameters (semen volume, sperm concentration, sperm total number, sperm motility and sperm
morphology) and redox parameters in blood, leukocytes, spermatozoa and seminal fluid.
RESULTS
Significantly lower levels of spermatozoa total count, concentration and motility were found in infertile patients
compared to control subjects. Blood leukocyte subpopulations from infertile patients displayed significantly increased
Reactive Oxygen Species (ROS) production. In plasma lipid peroxidation markers were significantly increased and
total antioxidant capacity (TAC) resulted significantly reduced compared to controls. Moreover, infertile men showed
significantly higher spermatozoa ROS levels and seminal plasma lipid peroxidation together with lower seminal fluid
TAC. All these data are indicative, in infertile patients, for a condition of oxidative stress in blood which is mirrored
in seminal fluid.
As regards correlation analyses, alterations in routine semen parameters significantly correlated with sperm ROS
production and seminal plasma lipid peroxidation suggesting a ROS-mediated spermatozoa dysfunction mechanism.
A positive and significant correlation between blood leukocyte ROS production, seminal plasma lipid peroxidation
and sperm ROS production was observed. A negative correlation was also observed between blood leukocyte ROS and
seminal plasma TAC.
CONCLUSIONS
Our results indicate the involvement of oxidative stress in sperm dysfunction, witness that systemic blood oxidative
stress reflects semen oxidative status and suggest blood leukocyte ROS estimation as a new potential and less invasive
indicator for male preconception care, especially for idiopathic infertile patients.
Endocrinology
P0996
VITAMIN D STATUS IN CHILDREN WITH GROWTH HORMONE DEFICIENCY
S. Tombari 1, Y. Amri 1, Y. Salem 2, Y. Hasni 2, R. Dabboubu 1, S. Ferchichi 3, t. Messaoud 1

1
Biochemistry Laboratory (LR 00SP03), Children’s Hospital-Tunis, Tunisia.
2
Department of endocrinology, CHU Farhat Hached, Faculty of medicine of sousse
3
Faculty of pharmacy, monastir · Biochemistry A
BACKGROUND-AIM
Growth Hormone Deficiency (GHD) is a rare disease defined by a total or partial impairment of growth hormone’s
production. However, vitamin D deficiency is frequent and may be associated with numerous pathologies. Few studies
have investigated the association between GHD and vitamin D deficiency. The aim of this study is to determine vitamin
D status in GHD patients and healthy subjects.
METHODS
A case control study was conducted at Children’s hospital of Tunis in collaboration of Farhat Hached’s hospital of
Sousse, included 39 GHD patients and 48 healthy subjects. Vitamin D was quantified using a chemiluminescence assay
and a full blood analysis was done. The statistical analysis was accomplished using SPSS.
RESULTS
Our study showed a significant increase of vitamin D concentration in GHD patients compared to healthy subjects
(p=0,049) and a significant decrease of calcium concentration in the GHD group (p=0,018). No significant difference
was found in phosphate concentration (p=0,62), ALP activity (p=0,65) and PTH level (p=0,15) between the two groups.
An interaction was noticed between vitamin D and ALP activity which was decreased in group with a vitamin D
concentration <30ng/mL (p=0,022). PTH concentration was negatively correlated to vitamin D concentration (r=-0,250,
p=0,021).
CONCLUSIONS
Our study showed that hypovitaminosis D is frequent in both patients and healthy subjects. In fact, GHD patients
seemed to be at a high risk of hypovitaminosis even when treated with rhGH, highlighting the importance of vitamin
D supplementation during treatment.
Endocrinology
P0997
ROLE OF THE CLINICAL LABORATORY IN THE DIAGNOSIS OF A PROLACTINOMA: A CASE REPORT
A. Perojil Jiménez 1, J.M. Villa Suárez 1, M.T. González Cejudo 1, F.J. Villanova Ruíz 1, T. De Haro Muñoz 1
1
Laboratorio de Análisis Clínicos, Hospital Universitario Clínico San Cecilio, Granada, Spain
BACKGROUND-AIM
Prolactinomas are the most common pituitary hormone-secreting tumors, being more frequent in women between
20-50 years with a gender ratio of 10:1. In women, most are microadenomas (<10 mm), presenting with the triad
amenorrhea, galactorrhea, and infertility. In men, most are macroadenomas (>10 mm) causing decreased libido,
erectile dysfunction, or gynecomastia.
METHODS
A 51-year-old man with no medical history who attended a Primary Care consultation reporting chronic insomnia and
erectile dysfunction. At the time of the consultation, he denies any use of medication.
RESULTS
In a first analysis, the results of Prolactin stood out: 863 ng/mL (2.6-13.1 ng/mL) and Testosterone: 130 ng/dL
(150-684 ng/dL) with a normal TSH value. Given the high suspicion of the presence of a prolactinoma, the patient is
treated early with Dostinex®, lowering his prolactin level in a subsequent determination to 125 ng/mL. A subsequent
contrast-enhanced magnetic resonance imaging that confirmed the presence of a 23mm pituitary macroadenoma that
remodeled the sella turcica and began to invade the right cavernous sinus.
CONCLUSIONS
Hyperprolactinemia is one of the most common neuroendocrine disorders, which consists of elevated serum
prolactin levels whose cause may be physiological (stress, insomnia or exercise), pharmacological (dopamine
receptor antagonists or antidepressants) or pathological (prolactinoma, hypothyroidism, chronic renal failure and
liver cirrhosis). One of the most important stimuli in its release is the thyrotropin-releasing hormone, which explains
why in patients with primary hypothyroidism there is a joint increase in this hormone as well as in prolactin.
Therefore, it is recommended to rule out primary hypothyroidism in people with hyperprolactinemia. Clinical guidelines
recommend that prolactin determination in a single extraction is sufficient for the diagnosis of hyperprolactinemia if
the venipuncture has not been traumatic. Although the gold standard continues to be magnetic resonance imaging,
the laboratory can be oriented towards diagnosis, taking into account that prolactin concentrations detected between
25-100 ng/mL may indicate a probable increase due to a functional cause, but concentrations higher than 200 ng/mL
indicate the presence of a prolactinoma as the probable cause.
Endocrinology
P0998
PREVALENCE OF VITAMIN D INTOXICATION IN A TERTIARY HOSPITAL
S. Sánchez Asís 1, P. Argente Del Castillo Rodríguez 1, B. García García 1, D. Morell García 1, M.I. Pastor García 1, A. Barceló
Bennasar 1
1
Department of Laboratory Medicine, Hospital Universitario Son Espases, Palma de Mallorca
BACKGROUND-AIM
Vitamin D is a fat-soluble hormone important for the intestinal absorption of calcium and to induce bone resorption.
It is synthesized mainly by the skin and only 10% is derived from dietary sources. The high prevalence of vitamin
D deficiency is currently a global problem that has led to an increase in vitamin D supplementation intake in the
general population. Consequently, this has contributed to a global increase in cases of vitamin D toxicity. Symptoms
of acute intoxication are mainly due to hypercalcemia and include confusion, polyuria, polydipsia, anorexia, vomiting
and muscle weakness. Also, chronic intoxication may cause nephrocalcionsis and bone demineralization.
METHODS
Retrospective study in a third-level hospital conducted between 2017 and 2021, in which prevalence in primary health
care and hospitalized patients with vitamin D concentrations above 100ng/mL was studied. From this patients, those
in which calcium concentrations were also determined were analyzed. Serum levels were measured using the Alinity
ci platform (Abbott, USA).
RESULTS
Vitamin D above 100ng/mL represented 0.2% of all vitamin D determinations during this period, with 579 cases.
In 100 patients of 579, calcium concentrations were determined;
From these, 75% of determinations in our population corresponded to women and 25% to men, with a mean age of
68 years old.
Additionally, 27% of the patients had vitamin D concentrations above 150ng/mL and 21% showed calcium serum
concentrations above reference values (>10.2 mg/dL).
Regarding the distribution of the analytical requests; 42% were derived from primary health care, 43% from out-patient
services and 15% from hospitalized patients.
CONCLUSIONS
The increase in supplementation with vitamin D and the rate of patients with high calcium concentrations highlights
the need to study vitamin D concentrations in supplemented patients.
In addition, the fact that the highest percentage of patients studied are women along with the high average age,
indicates a greater follow-up in menopausal women and, consequently, other supplemented patients might not be
being studied.
Although this percentage of vitamin D intoxication might be low it represents a large amount of patients in absolutes
numbers.
Endocrinology
P0999
PERFORMANCE OF AT-HOME SELF-COLLECTED SALIVA SAMPLING IN THE MONITORING OF CORTISOL LEVELS IN
ENDOCRINE DISEASES SUCH AS CUSHING’S SYNDROME
M. Rogozinska 2, R. Rola 1, K. Kowalski 1, T. Bienkowski 1

1
Masdiag - Diagnostic Mass Spectrometry Laboratory, 33 Stefana Zeromskiego St. PL - 01-882 Warsaw, Poland
2
Masdiag - Diagnostic Mass Spectrometry Laboratory, 33 Stefana Zeromskiego St. PL - 01-882 Warsaw, Poland; Faculty of
Chemistry, University of Warsaw, 1 Pasteura St., PL - 02-093 Warsaw, Poland
BACKGROUND-AIM
Cortisol is a glucocorticoid class hormone which plays a major role in regulating a variety of physiological processes.
Monitoring of its level is commonly used in the assessment of adrenal, pituitary and hypothalamic function and is
especially important in patients with endocrine diseases such as Cushing’s syndrome (CS). The Endocrine Society
Clinical Practice Guideline includes late night salivary cortisol (LNSC) as an initial testing for CS. Due to the fact that
salivary free cortisol is in an equilibrium with plasma free cortisol, LNSC seems to be an accurate method to evaluate the
level of its biological active form. In Poland, there are about 1,000 patients diagnosed with CS spread evenly throughout
the country, who need permanent monitoring of cortisol levels, especially during cortisol-lowering therapy. Since saliva
collection does not require special equipment, even non-professionals can easily collect it themselves at home without
a need for hospitalization. The aim of our study was to perform an at-home, self-collected saliva sampling with the
usage of the Cortisol-Salivette® saliva collection system to control cortisol levels in patients with CS.
METHODS
The methodology assumptions require firstly, storage of non-centrifuged saliva collectors at home and secondly their
transport to the laboratory. Therefore, the key parameter in assessing its usefulness is the determination of salivary
cortisol stability collected on the swab in order to simulate at-home storage and shipping conditions.
RESULTS
Stability studies show that storage of non-centrifuged collectors at 4-8°C and at room temperature (RT) has no
significant effect on cortisol concentration up to 14 days. At 4-8°C the concentration levels spread in the range of
92%-100%, while at RT in the range of 86%-105%. This time is sufficient for sample delivery to laboratory and its
analysis. It is noteworthy, that centrifugation and freezing extend this time even to more than a month. Repeated
freeze-thaws cycles do not adversely affect the cortisol levels.
CONCLUSIONS
In conclusion, LNSC could be a simple, cheap, non-invasive method, compares very favorably with alternative screening
tests, which together with sensitive LC-MS/MS method could provide a reliable tool in at-home monitoring cortisol
levels in patients with CS.
Endocrinology
P1000
COMPARATION BETWEEN TWO-STEP AND ONE-STEP TEST FOR DIAGNOSIS OF GESTATIONAL DIABETES
R. Sansano Galiano 1, I. Cebreiros López 1, R.P. Cano Mármol 3, M. Fernández López 2, P.J. Sánchez Carretero 1, C. Guirao
Blázquez 1, S. Requena López 1, J.A. Noguera Velasco 1
1
DEPARTMENT OF CLINICAL ANALYSIS, VIRGEN DE LA ARRIXACA UNIVERSITY CLINICAL HOSPITAL, MURCIA.
2
DEPARTMENT OF ENDOCRINOLOGY AND NUTRITION, VIRGEN DE LA ARRIXACA UNIVERSITY CLINICAL HOSPITAL, MURCIA
3
DEPARTMENT OF ENDOCRINOLOGY AND NUTRITION, VIRGEN DE LA ARRIXACA UNIVERSITY CLINICAL HOSPITAL, MURCIA.
BACKGROUND-AIM
Gestational diabetes (GD) is defined as any intolerance grade to glucose which is detected for the first time during
pregnancy. That is associated with an increased morbidity and mortality for patient and the fetus in both short and
large term. For that reason, is important its detection, control and treatment.
Diagnosis consists on a two-step test. First, patients must take 50 g of glucose and then, a measurement of serum
glucose is performed one hour later. If glycaemia is less than 140 mg/dL patient is discarded. Otherwise, patient must
undergo a second test: take 100 g of glucose and measurement of glycaemia at 0, 1, 2 and 3 hours, being cut points
105, 190, 165 and 145 mg/dL, respectively. If two points are positives, it is considered GD diagnosis (gold standard
(GS) method).
At 2010, the International Association of the Diabetes and Pregnancy Study Groups (IADPSG), inspired by a study of
Hyperglycaemia and Adverse Pregnancy Outcome at 2008, proposed one-step diagnosis in order to reduce the quantity
resources and costs. The algorithm is based on the intake 75 g of glucose and glycaemia determination at 0, 1 and 2
hours. If one point is positive, GD is confirmed. Cut points are 92, 180 and 153 g/dL, respectively.
The aim of this study is to compare the GS criteria of GD with the IADPSG criteria in our population.
METHODS
GD curves realized on 'Virgen de la arrixaca' hospital in 2022 (2132 patients) were extracted and selected for basal
glucose greater than 91 mg/dL (130 patients, taken as patient population), considering those patients as GD positive
according to IADPSG criteria and were compared with GS diagnosis.
RESULTS
The number of patients diagnosed with GS method in the population were 39 and 130 with IADPSG criteria, which
mean an excess of 91 patients (which represents 70% of the population).
Among the total determinations (2132), 113 patients were diagnosed with GD by GS and 204 would have been
diagnosed by IADPSG criteria, which represents a PPV of 55% and a specificity of 95%.
CONCLUSIONS
According to data obtained, it is not possible to conclude that the change in diagnostic criteria is appropriated.
Nevertheless, we consider that each hospital should make these kind of decisions based on the total number of patients
undergoing the test considering the resources-cost ratio
Endocrinology
P1001
EVALUATION OF BLOOD ADRENOCORTICOTROPIC HORMONE AND CORTISOL LEVELS PRE AND POST-CORTICOSTEROID
TREATMENT IN COVID-19 PATIENTS WITH ACUTE RESPIRATORY DISTRESS SYNDROME
V.I. Mayasari 2, A. Aryati 2, H.E. Sidjabat 3, B.P. Semedi 1, F.R. Marpaung 2

1
Department of Anesthesiology and Reanimation Dr. Soetomo Academic Hospital/ Faculty of Medicine, Universitas
Airlangga, Surabaya, Indonesia
2
Department of Clinical Pathology Dr.Soetomo Academic Hospital/ Faculty of Medicine, Universitas Airlangga, Surabaya,
Indonesia
3
Menzies Health Institute Queensland, Griffith University, Queensland, Australia
BACKGROUND-AIM
Measurement of adrenocorticotropic hormone (ACTH) and cortisol levels becomes more important in the case of
corticosteroid therapy due to their impact on ACTH and cortisol secretion and the hypothalamic-pituitary-adrenal (HPA)
axis. However, clinical evidence on the usefulness of monitoring ACTH and cortisol in Covid-19 patients, especially those
with Acute Respiratory Distress Syndrome (ARDS), still needs to be provided. Therefore, this study aimed to analyze
ACTH and blood cortisol levels in a cohort of COVID-19 patients with ARDS who received corticosteroid therapy.
METHODS
This research was a prospective cohort study of 45 Covid-19 ARDS patients treated in the ICU Isolation of Dr. Soetomo
Academic Hospital. Since 15 subjects dropped out on the seventh day of follow-up, samples for analysis of ACTH and
cortisol levels before and after corticosteroid administration were obtained from 30 subjects who could be followed
up on the seventh day. Plasma ACTH levels and morning serum total cortisol were measured at treatment day one and
day 7 in 30 patients receiving corticosteroids. The correlation between ACTH and cortisol levels with patient outcomes
was based on survival at day 28.
RESULTS
There was a significant difference between ACTH and cortisol levels before and after corticosteroid administration
( 2.63 pg/mL and 28.25±16,839 µg/dL, respectively). The was no correlation between ACTH levels after corticosteroid
administration and patient outcome, while cortisol was moderately associated (p=0.002). There were significant
differences in cortisol levels after administration of corticosteroids based on outcomes between the survive (mean
22.99±15,428 µg/dL) and the non-survive (36.15±16,317 µg/dL)
CONCLUSIONS
There was a significant difference between ACTH and cortisol levels before and after corticosteroid administration
in Covid-19 patients with ARDS, where ACTH and cortisol levels after corticosteroid administration are significantly
higher. Furthermore, there was no relationship between ACTH levels after corticosteroid administration and the
outcome of Covid-19 patients with ARDS. However, there was a positive correlation between cortisol levels after
corticosteroid administration and the outcomes of Covid-19 patients with ARDS, where higher cortisol levels were
found in the deceased patients.
Endocrinology
P1002
PATTERN OF THYROID FUNCTION TEST REQUEST AMONG PAEDIARICS WITH DOWN SYNDROME IN A TERTIARY HEALTH
INSTITUTIONS’ TUMOR MARKER LABORATORY IN NIGERIA: A TWO YEAR REVIEW
a. Azeez 1
1
university college hospital
BACKGROUND-AIM
This study is a two year review of requests pattern for thyroid function test(tft) among paediatrics, in a Nigerian tertiary
health care institution. Prostate specific antigen was first described about 44 years ago but is still in use today for,
diagnosis, monitoring, screening and prognosis of prostatic carcinoma though not-very specific as was widely believed.
Prostate cancer is an increasingly important public health problem among adult men worldwide. Nigeria, which was
formerly regarded as a low-incidence area by several authors is now witnessing a steep rise in the occurrence of this
disease. This has been suggested to be due to increasing availability of screening tests and diagnostic facilities and
not necessarily because of increased incidence of the diseases. Many notable Nigerians have died due to this dreaded
disease
METHODS
All plasma samples for PSA from January 2021-December 2022 were analyzed weekly by abbot autoanalyser,
chemiluminescence assay method. Bio-data from request form were collated and analyzed. A total of 385 requests
were received for the period under review.
RESULTS
There was an increase of request from inception to the last year of review. Smoked or barbecued food, consumption of
local herb and alcohol in order of importance, respectively, appears to be prominent factor in patient requested for PSA.
The mean age was 67.years; the youngest was 29, while the oldest was 93years. Age 70 has the highest frequency of
8.5% .Mean PSA was 12.9ng/ml. There was a positive correlation between age and PSA (R = 0.255, P < 0.05).Significant
increase in PSA with age were reported. Men who retired from active jobs constitute the highest request for PSA test
CONCLUSIONS
There was an increasing trend in the proportion of requests with values outside the reference range especially in
patients diagnosed of BPH, prostate cancer, while some routine test for PSA were elevated for the first time .This is
in line with earlier report of increasing incidence of prostate cancer in Nigeria despite the increasing knowledge of
healthy lifestyle
Endocrinology
P1003
HYPERTHYROIDISM SECONDARY TO GESTATIONAL TROPHOBLASTIC DISEASE: A CASE REPORT
A. Blanco 1, S. Molina 1, J. Sánchez 1, A. Pascual 1, H.A. Sanz 1, B. Beteré 1, L. Maceda 1, K. Sidak 1, C. Tenreiro 1, A.M.
Moreno 1, Y. Fernández 1, M. De La Cabeza 1, S. Lapeña 1, M. Barrionuevo 1, F. Álvarez 1, M.J. Ruiz 1, A. Rodríguez 1, A.
Sánchez 1, L. Varela 1, M.J. Álvarez 1, J.M. Gasalla 1
1
Clinical Analysis Service, Principe de Asturias University Hospital, Alcala de Henares
BACKGROUND-AIM
Gestational trophoblastic disease (GTD) corresponds to a spectrum of proliferative lesions of trophoblastic tissue
that can occur during pregnancy: hydatidiform mole (HM), choriocarcinoma, trophoblastic tumor and epithelioid
trophoblastic tumor. All these entities are characterized by hypersecretion of human chorionic gonadotropin hormone
(hCG), with its levels being directly proportional to the tumor mass.
hCG is a glycoprotein composed of an alpha subunit common to other hormones, including thyroid-stimulating
hormone (TSH). When hCG levels remain very high (>100000 mU/mL) for several weeks, it is able to weakly stimulate
thyroid hormone production by binding to its receptor on the thyroid gland and producing a thyrotropic effect.
METHODS
We present the case of a 41-year-old woman with amenorrhea of 7 weeks who comes to the emergency department for
vaginal bleeding of 4 days of evolution. The urine pregnancy test was positive and the transvaginal ultrasound showed
a uterine mass of 11x9 cm, compatible with the diagnosis of HM.
RESULTS
Among the laboratory parameters, serum β-hCG of 420588 mU/ml, TSH 0.04 µU/ml (normal range 0.4-5 µU/mL), T4
1.68 ng/dL (normal range 0.89-1.76 ng/dL) and T3 5.15 ng/dL (normal range 2.3-4.2 pg/mL) stand out. A curettage was
performed for biopsy where the results confirmed abortive remains and morphological alterations compatible with
complete HM.
A laparoscopic hysterectomy was scheduled and performed in the emergency department 9 days later due to more
abundant bleeding, and follow-up was done over time with β-hCG levels:
Day 0: 420588 mU/mL
Day 9 (post-hysterectomy): 21977 mU/ml
Day 10: 17852 mU/mL
Day 18: 2478 mU/mL
On day 18 post-hysterectomy, thyroid function is also monitored, which is corrected with the resolution of the HM
(TSH: 1,471 mU/mL).
CONCLUSIONS
In the case presented, despite the high β-hCG concentration and suppressed TSH levels, the patient does not develop
hyperthyroid symptoms. However, it demonstrates the importance of suspecting possible hyperthyroidism in any
patient with GTD for early treatment.
Endocrinology
P1004
A NEW RAPID CORTISOL ASSAY FOR THE INTRAPROCEDURAL MONITORING DURING ADRENAL VEIN SAMPLING
INTERVENTION
M. Taglieri 1, L. Di Gregorio 1, B. Donati Marello 1, A. Di Guida 1, P. Merlach 1, T. Manetta 1, G. Mengozzi 1

1
Clinical Biochemistry Laboratory, AOU Città della Salute e della Scienza di Torino, Torino, Italy.
BACKGROUND-AIM
Adrenal vein sampling (AVS) procedure should be considered the gold standard for the management of patients with
primary aldosteronism, the most common form of secondary arterial hypertension. Technical difficulties with this
procedure can be overcome by monitoring cortisol concentrations at the different venous sites.
The objective of this study is to evaluate the usefulness of the rapid determination of cortisol by a recently developed
method directly in the interventional radiology room.
METHODS
The new rapid assay (NBCL, Nijmegen, The Netherlands; distributed by Pantec Srl) is designed to measure whole
blood cortisol concentrations on samples collected in EDTA-containing tubes. The time to result is 5 minutes, limit of
detection is 6 µg/dL with an assay range up to 2000 µg/dL. A precision study using two controls (10.4 µg/dL and 53.3
µg/dL) resulted in a total CV of 8.7% and 5.9%, respectively.
RESULTS
Although AVS protocols are not yet standardized and can vary widely among centers, to evaluate the cannulation
success we consider that the ratio of cortisol concentration in the adrenal vein and the vena cava should be over 3.
Within our study, the success of the procedure was confirmed in eight out of nine patients examined. The Passing-
Bablok regression between rapid and routine (Abbott Diagnostics) assay yielded Abbott=4.39+0.70NBCL, with a
coefficient of correlation of 0.67 (95% CI 0.45-0.81). Bland-Altman plot showed a mean percentage difference of -20.5%
(95% CI 42.1-1.00) with a wide dispersion of data.
CONCLUSIONS
The new approach appeared reliable for the timing intraprocedural monitoring of cortisol. There are some precautions
that should be taken into account. It is important to turn on the instrument about 30 minutes before the start
of the procedure and make sure that the whole blood sample is properly mixed and free of foam and clots. In
addition, it is advisable to pay attention to the transport of the instrument avoiding inappropriate movements. Our
correlation analysis with the routine assay makes results obtained with NBLC cortisol not interchangeable for the final
interpretation of the data.
Endocrinology
P1005
FUNCTIONAL AND STRUCTURAL CHARACTERIZATION OF THE GENETIC VARIANT P.PRO292LEU IN A PATIENT WITH
ADULT HYPOPHOSPHATASIA
J.M. Villa Suárez 2, T. González Cejudo 2, A. Perojil Jiménez 2, B. García Fontana 1, M. Muñoz Torres 1, T. De Haro Muñoz 2
1
Instituto de Investigación Biosanitaria de Granada, Granada
2
Laboratory Clinical Management Unit, Hospital Universitario Clínico San Cecilio, Granada
BACKGROUND-AIM
Hypophosphatasia (HPP) is an ultra-rare genetic disease caused by one or more mutations in the ALPL gene encoding
tissue non-specific alkaline phosphatase (TNSALP). HPP has a high clinical variability associated primarily with the
accumulation of enzyme substrates, including pyridoxal-5'-phosphate (PLP). The main clinical manifestations include
defects in bone and dental mineralization, pain, muscle weakness, or renal abnormalities.
METHODS
A 62-year-old man who consulted for knee pain and meniscopathy of years of evolution, with no history of fractures,
chondrocalcinosis, or dental abnormalities. A persistent decrease in alkaline phosphatase activity and very high PLP
levels relative to reference intervals were observed. After ALPL gene sequencing, it had the c.875C>T variant; p.
(Pro292Leu) in exon 9 in heterozygosity. To determine the degree of involvement of the enzyme and perform a
genotypic-phenotypic and structural linkage of the variant, functional characterization studies of this TNSALP genetic
variant were performed. For this, human embryonic kidney cells (HEK293T) were transfected with pcDNA 3.1 vectors
containing the ALPL gene variant to be studied and the WT-ALPL sequence by lipofection using the LipoD293 reagent.
Subsequently, the alkaline phosphatase activity of each of the cultures was determined. To predict the effect of the
variant on the structure of TNSALP, a 3D model based on the sequence homology between TNSALP and the placental
PALP isoenzyme was obtained.
RESULTS
Cells transfected with the genetic variant under study showed a drastic decrease in alkaline phosphatase activity with
respect to WT-ALPL. Regarding the structural analysis of the protein, the p.Pro292Leu variant seems to affect the
structural domain involved in calcium binding. Calcium binding is crucial for the correct folding and assembly of the
TNSALP molecule and therefore appears to be essential for its activity in bone mineralization.
CONCLUSIONS
The variant identified in our patient p.(Pro292Leu) gives rise to a mild phenotype of adult HPP. Initially, the p.
(Pro292Leu) variant was classified according to ACMG recommendations as a variant of uncertain significance.
However, upon reassessment, it has now been classified as probably pathogenic, consistent with the results observed
in our study.
Endocrinology
P1006
HEADACHE AND MACROPROLACTINOMA: A CASE REPORT
A. Pérez Rodríguez 1, N. López Lazareno 1, A. Verdejo González 2, J. Romero Torres 2, A. Guillamón Seoane 2, D. Martín
Fernández 2, L. Prieri 2, V.d.A. Moral Ortiz 2, D. Navarro Calderón 2, L. Marques De Brito 2, A. Bravo Gómez 2, A.E. Zamora
Trillo 2, E. Llorente Martín 2, S. Sánchez Berdial 2, M. García Gámiz 2
1
Gregorio Marañón University General Hospital
2
Gregorio Marañón University General Hospital
BACKGROUND-AIM
A 33-year-old man, who has been suffering from headaches for the past 2 years, was referred to Neurology Service. He
refered pain in the frontal region, which sometimes radiates bilaterally to the occipital region, predominantly on the
right side. It was an oppressive type pain that occured when coughing, sneezing, bending over or at the beginning of
the exercise. The duration of the pain was 1-2 seconds. There were not migraine, photo/sonophobia, rhinorrhea, nausea
or vomiting. Confrontation perimetry, fundus and deep-tendon reflexes were normal. The patient was established to
have exertional headache.
METHODS
A study of the pituitary gland was requested with imaging and laboratory tests that included measurement of the main
pituitary hormones. TSH, FSH, LH, prolactin, testosterone were measured in Alinity i; ACTH, GH, IGF1 were measured in
Liaison. Both kits use microparticle chemiluminescence (CLIA).
RESULTS
Imaging test:
Magnetic resonance angiography showed a 22x19.5x18mm sellar mass compatible with a right pituitary
macroadenoma, with slight suprasellar extension.
Blood analysis:
TSH 1.47mIU/L(0.35-4.94); ACTH 21.7ng/L(5.0-60.0); GH 0.10µg/L(0.10-5.00); IGF-1 232µg/L(113-250); SDS IGF-1
Calculation regarding age 1,785(-2 to +2); FSH 2.5IU/L; LH 1.2IU/L; Prolactin 4616.9µg/L(3.5-19.4); Testosterone 2.8µg/
L( 2.4-8.7); Cortisol 8.8µg/dL(5.0-25.0). The biochemistry test was normal except for a slight elevation of uric acid and
calcium.
The presence of a prolactin-producing pituitary macroadenoma (macroprolactinoma) was identified and cabergoline
was prescribed. After 2 months, the patient is feeling better and is awaiting analytical and imaging tests to see the
evolution of the mass found.
CONCLUSIONS
Headaches are generally tensional or migraineos, but as we have seen, they can be associated with pituitary adenoma
due to the compression they exert on adjacent regions. These adenomas may be related to endocrinopathy if they
lead to an increase in the production of some hormone, being prolactin the most frequent, or a decrease of them
(hypopituitarism). Imaging and laboratory tests help differential diagnosis as the first ones show location and
extension of the mass and the second ones indicate functionality. Laboratory tests are key to establish the best
treatment for the patient.
Endocrinology
P1007
INSULIN LIKE GROWTH FACTOR-1, HUMAN GROWTH HORMONE AND TESTOSTERONE AS POTENTIAL BIOMARKERS IN
LABORATORY DIAGNOSIS AND PROGNOSIS OF AFRICAN BLACKS BEFORE AND AFTER BILATERAL ORCHIDECTOMY
A. Oniye 1, M. Charles- Davies 1, O. Olaopa-Olapade 2, J. Anetor 1, O. Agbedana 1

1
Department of Chemical Pathology, College of Medicine, University of Ibadan, Ibadan
2
Department of Surgery, College of Medicine, University of Ibadan, Ibadan.
BACKGROUND-AIM
Mortality is high in African men, who are at high risk for castrate resistant Prostate Cancer (PCa). Bilateral ochidectomy
(BO) is the most preferred treatment option due to late presentation and advanced disease. Prostate specific antigen
(PSA) has limited value in laboratory diagnosis of PCa. Elevated Insulin like growth factor-1 (IGF-1), human growth
hormone (hGH) and testosterone levels appear to increase PCa risk. The importance of IGF-I in the early detection of
PCa has been suggested and is therefore investigated in this study.
METHODS
142 male participants aged 50-90 years were enrolled into this prospective cohort study in the Urological Clinic of a
teaching hospital in Ibadan and environs. PCa (n=53) group was age matched with benign prostate hyperplasia (BPH,
n=45) and Control (apparently healthy men, n=44) groups. PCa and BPH were diagnosed histologically while BO was
performed among PCa group only (BOP, n=22). Serum obtained from fasting blood (5mL) was obtained for HGH, IGF-1,
testosterone and PSA estimation by enzyme linked immunosorbent assay. All indices were assessed at baseline and 12
months after BO in BOP only. Data analysed using Student’s t-test and multiple regression were considered significant
at p<0.05.
RESULTS
The levels of PSA, hGH and IGF-1 were significantly higher in PCa than BPH and ControI groups (p<0.05). Only PSA level
was significantly elevated when BPH group was compared with Control (p<0.001). Testosterone levels were however,
similar in all groups at baseline. After BO, testosterone and IGF-1 levels reduced while the PSA and hGH levels increased.
At baseline, mortality (28%) observed in PCa group only was associated with increased hGH and reduced testosterone
levels (p= 0.002, 0.025 respectively). There were also significantly positive relationships among PSA, IGF-1 and hGH
(p=0.003).
CONCLUSIONS
IGF-1 may be considered as a better potential prognostic laboratory biomarker than hGH and testosterone in males
with PCa.
Endocrinology
P1008
DETERMINATION OF THE CUTOFF CORTISOL CONCENTRATION FOR THE DIAGNOSIS OF ADRENAL INSUFFICIENCY AFTER
ACTH STIMULATION USING THE ROCHE ELECSYS CORTISOL II ASSAY
H. Poplašen 1, M. Zupančič 1
1
Department of Laboratory Diagnostics, Institute of Oncology, Ljubljana
BACKGROUND-AIM
Assessing serum cortisol levels is crucial in the diagnosis and monitoring of patients with adrenal dysfunction. An
accurate diagnosis requires measuring basal cortisol values and cortisol levels following synthetic adenocorticotropic
hormone (ACTH) stimulation and interpretation of results based on established cutoff values. In our laboratory, we
have started using the Roche Elecsys Cortisol II assay. This assay is standardized with the IRMM/IFCC-451 panel,
using the ID-GC/MS. It uses monoclonal antibodies with lower cross-reactivity to other steroids, which results in lower
measured cortisol concentrations. Currently used guidelines state cutoff value of 500 nmol/L or higher, which are not
appropriate for this assay. Therefore, we have done an inter-laboratory comparison using Roche Elecsys Cortisol II and
Siemens Atellica IM Cortisol to establish new cutoff value.
METHODS
The serum cortisol levels were measured in 130 samples of various patients using the Siemens Atellica (Atellica IM
Cortisol) and Roche Cobas e801 (Roche Elecsys Cortisol II), both using the competitive immunochemical method
with chemiluminescent detection. We compared and statistically processed the results in the MedCalc program, using
comparative analysis with Passing-Bablok, Bland-Altman and ROC analysis.
RESULTS
On average, Roche Elecsys Cortisol II gave 20-23% lower cortisol concentrations than Siemens Attelica IM Cortisol.
Using ROC analysis, we obtained an optimal cortisol cutoff value of 392 nmol/L (diagnostic sensitivity 100%, diagnostic
specificity 96.6%). Using Passing-Bablok regression, we calculated a cutoff value of 397 nmol/L. Both results are
consistent with data from literature.
CONCLUSIONS
Based on the results obtained, we concluded that the currently established cutoff value for cortisol measurements
following ACTH stimulation are not suitable for interpretation with our assay, as it would result in a high percentage
of false positive diagnosis of adrenal insufficiency for our patients. A new cutoff value of 392 nmol/L (in agreement
with doctors, the value was rounded to 400 nmol/L) is used in our institution for assessment of adrenal function with
Roche Elecsys Cortisol II assay. In the future, we plan to optimise this study using only samples of our patients.
Endocrinology
P1009
CHILDHOOD SEXUAL ABUSE IS ASSOCIATED WITH HIGHER TOTAL GHRELIN SERUM LEVELS IN ADULTHOOD: RESULTS
FROM A LARGE, POPULATION-BASED STUDY
D. Wittekind 5, K. Jürgen 5, M. Roland 3, K. Wirkner 4, R. Baber 5, C. Sander 2, V. Witte 1, A. Villringer 1, M. Kluge 2

1
Clinic of Cognitive Neurology, University of Leipzig, and Department of Neurology, Max Planck Institute for Cognitive and
Brain Sciences, Leipzig, Germany
2
Department of Psychiatry and Psychotherapy, University of Leipzig, Leipzig, Germany
3
Institute of Psychology, Universität der Bundeswehr München, Neubiberg, Germany
4
Leipzig Research Center for Civilization Diseases (LIFE), University of Leipzig, Leipzig, Germany
5
University of Leipzig, Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics
BACKGROUND-AIM
Ghrelin is an orexigenic peptide hormone synthesized in times of stress and hunger and alterations of the ghrelin
system following acute stressors could be repeatedly shown in himans. However, little data exists on long-term effects
of trauma on the ghrelin system. We aimed to investigate the influence of childhood trauma on total ghrelin serum
levels in a large, population-based study
METHODS
Total serum ghrelin was measured in 1666 participants of a population-based cross-sectional study (‘LIFE study’). The
Childhood Trauma Screener (CTS) was used for the assessment of childhood trauma in the final sample (N=1086; mean
age: 57.10 ± 16.23 years; 632 males, 434 females). Multiple linear regression analyses and generalized linear models
were chosen to examine the association between childhood trauma and total serum ghrelin concentrations.
RESULTS
Childhood sexual abuse went along with significantly higher ghrelin serum levels in the total sample (β = 0.114, p =
0.00008) and in women (β = 0.142, p = 0.002), but not in men (β = 0.055; p = 0.166). Women with severe emotional
neglect in the childhood had significantly higher ghrelin levels than those without (odds ratio = 1.204; p = 0.018). For
the CTS Sum Score and other CTS sub-scale scores, no significant association with ghrelin serum levels was found.
CONCLUSIONS
Our study is the first to show associations between childhood sexual trauma and total ghrelin levels in adults in a large,
community-based sample. Our results should initiate further research of the role of ghrelin in human stress response
in prospective study designs.
Endocrinology
P1010
PRESENCE OF MACROPROLACTIN IN ADULT POPULATION OF KOSOVO
M. Kurshumliu 1, I. Ibrahimi 2, A. Zeka 3

1
Prolab
2
Heimerer College
3
Prolab
BACKGROUND-AIM
Hyperprolactinaemia is generally diagnosed by laboratory testing for elevated prolactin concentrations. However, not
all subjects with high prolactin levels show classical symptoms of hyperprolactinaemia. This is mainly due to presence
of macroprolactin, a complexed form of prolactin with auto IgG antibodies. Although, macroprolactin is detected in
routine laboratory tests for hyperprolactinaemia, it is inactive, thus giving falsely elevated results for prolactin which
can lead to false diagnosis of hyperprolactinaemia.
Our study aims to assess the prevalence of macroprolactinaemia in adults in Kosovo in patients tested for
hyperprolactinaemia and a sample of the population.
METHODS
Macroprolactin concentration was determined using polyethylene glycol (PEG) precipitation. 69 female and 34 male
subjects were tested. Female subjects were mainly found to have increased prolactin levels. Male subjects were
randomly selected. We analysed overall levels of prolactin in subjects, and the percentage macroprolactin in prolactin
concentrations. We also compared prolactin and macroprolactin levels between male and female subjects using the
Welch t-test for unequal variance.
RESULTS
Our study demonstrates that macroprolactin is present in most samples, in varying ratios, along with the monomeric
(true) form of prolactin. From the group, 14.5% of female and 11.4% of male subjects have clinical macroprolactinaemia
(macroprolactin > 65%). In addition, most of the samples (n= 86 from 104) had a macroprolactin percentage in the
equivocal range (20-65%), which demonstrates the tendency of prolactin to be bound to auto IgG antibodies. Prolactin
levels were significantly higher in female subjects (p<0.05) compared with male subjects. For both genders, we found
that the oprolactin levels were significantly higher before precipitation to remove macroprolactin.
CONCLUSIONS
The data obtained have clearly demonstrated the presence of macroprolactinaemia, of which many laboratories and
clinicians may not be aware. Therefore, increased awareness for the interference of macroprolactin in total prolactin
concentration is needed when diagnosing hyperprolactinaemia.
Endocrinology
P1011
ARE THESE DIFFERENCES BETWEEN ANDROGENS WHEN STUDYING ANDROGENISATION IN THE TRANSGENDER
PATIENT?
G. Velasco De Cos 1, C. Latorre Mesa 2, R.J. García Martinez 1, P. Martín Audera 1, F. Pons Vidal 1, N.H. Cahuana Santamaría
1
, A. Villar Bonet 1, M.T. García Unzueta 1
1
Hospital Universitario Marqués de Valdecilla
2
Hospital Universitario Marqués de Valdecilla, Santander, Spain
BACKGROUND-AIM
In gender affirming therapy of the transgender male, increasing doses of testosterone are administered and as the
androgen dose increases, various biochemical parameters such as haematocrit, creatinine and PSA increase. It is
not known whether any parameters have advantages in monitoring this androgenisation. Our study evaluates the
correlation between the increase in 3 α androstanediol glucuronide, testosterone, free testosterone and FAI with the
aforementioned biochemical parameters in a cohort of trans men.
METHODS
Samples were obtained from 36 transgender men at the start of therapy and after each dose increase (3 months) during
one year of treatment. Of the 36, at the time of the study, 4 had received only 25mg/month doses, 4 with 50mg/month
doses, 8 with 100mg/month doses and 20 had completed the year and were at maximum doses (250mg/month).
RESULTS
For haematocrit all androgens analysed were significantly correlated, however there are differences in the Rho between
each pair. FAI showed the best correlation: ρFAI=0.525. The correlation coefficients for the other androgens were:
ρtestosterone=0.454, ρfTestosterone= 0.436, ρDHEAS=0.383, ρ3αAG=0.350.
For creatinine no significance was obtained, although 3αAG(p=0.081) and DHEAS(p=0.083) showed a clear trend
towards a significant correlation. When analysing only the data from the 20 TXM who had completed the year,
significance was found for DHEAS (p=0.010, ρ=0.297).
For PSA, as for haematocrit, all androgens were significant. The best correlation was for free testosterone
ρfTestosterone=0.431. The correlation coefficients for the other androgens were: ρTestosterone=0.403, ρ3αAG=0.380,
ρFAI=0.333 and ρDHEAS=256.
CONCLUSIONS
The different androgens obtain different correlation coefficients depending on the marker used. FAI obtained the best
results with haematocrit, making it the parameter of choice when assessing the relationship between a given dose
and its increase. For creatinine we did not obtain significance, but it is striking that in this parameter, unlike PSA and
haematocrit, the best results were obtained with testosterone metabolites.
Endocrinology
P1012
MOLECULAR CHARACTERIZATION OF RISK NON-HLA CODING VARIANTS IN THE PEDIATRIC POPULATION WITH TYPE
1 DIABETES
C. Perez Barrios 1, M. Diez Blanco 1, E. Donoso Navarro 1, N. Santiesteban 1, A. Royuela 1, E. Colino 1, F. Bernabeu 1, P. Ros 1
1
Puerta de Hierro Hospital
BACKGROUND-AIM
Type 1 diabetes mellitus (T1DM) is the most common chronic metabolic disease in children. Despite the fact that
its origin is multifactorial, it is known that genetics plays an important role in the autoimmunity described in these
patients, having identified numerous risk alleles. Of these, those located in the non-HLA coding region are the least
known, and may be involved in the symptoms of patients, being potential markers for targeted therapies.
METHODS
A total of 163 pediatric patients (<18 years) diagnosed with Type 1 Diabetes were recruited. DNA extraction was
performed using the QIAamp® DNA Mini Kit system (Qiagen) from K3-EDTA samples. Variants were analyzed with 4
pre-designed Taqman probes (C__16021387_20, C___2415786_20, C___1464836_20, C___2981072_10) and 1 custom-
designed probe (ANWDEJA) from Thermofisher in a StepOnePlus qPCR kit (Applied Biosystems). Clinical data was
collected from the electronic Clinical Record. Population frequency data were obtained from the reference SNP
Reporting (NIH) database.
RESULTS
A total of 163 patients were analyzed, of which 78 were women and 85 men. The median age of debut was 93
months [IQR: 53-131.75], with no significant differences by sex. Of the 153 patients with data collected for pancreatic
autoimmunity (aGAD, aIA2 and aIA), 144 had some positive antibody (94.11%) and 9 were negative for the 3
autoantibodies analyzed. On the other hand, of the 132 patients with information of thyroid immunity, 26 presented
autoantibodies (aTPO or aTG), 6 of them developed thyroid disease. In addition, 21 patients had celiac disease
autoantibodies (aTG and aTGt/aE), of which 11 were diagnosed with celiac disease.
All risk allele frequencies were higher than those described for the population (23.3% vs 8.6%, 56,4% vs 37.2% , 73.0
% vs 48.75, 74,8% vs 46.0% and 76.1% vs 47.1% for PTPN22, CTLA4, CD226, SH2B3 and FUT2). Furthermore, CTA4 risk
allele were related with celiac disease and GAD antibodies, whereas PTPN22 risk allele were related with IA2 antibodies.
CONCLUSIONS
Even though the results are preliminary, we observed that risk variants are clearly overrepresented in the study
population, in agreement with the high percentage of patients presenting pancreatic, thyroid, or celiac autoimmunity.
Endocrinology
P1013
LABORATORY DIAGNOSIS OF INSULINOMA, CASE REPORT
R. Arapi 1, B. Cullhaj 2
1
Genius Laboratory Network ,Department of Laboratory Medicine, University of Medicine, University Hospital Center "Mother
Teresa"
2
Laboratory Doctor and Department of Microbiology, M.D Microbiologist, Genius Laboratory Network, Albania
BACKGROUND-AIM
Insulinoma is a insulin secreting tumor. It is a rare tumor and the diagnose can be difficult and it can take a long time.
Insulinoma is usually located inside the pancreas but ectopic cases have been reported.
METHODS
Tests that determine the diagnose are high c-peptide, high insulin and high proinsulin following episodes of
hypoglycemia. C-peptide and insulin were performed with chemiluminescence immunoassay (CLIA) and enhanced
chemiluminescence immunoassay (ECLIA) respectively , proinsulin was performed with enzyme immunoassay (EIA).
RESULTS
Case report: A 48 year old female patient presents to the emergency room with tremor, sweating and mental fog. This
was her 4 visit to the ER in the last 2 month. The onset of symptoms was 1 year prior with tiredness and irritability
during the day .Her symptoms were treated as anxiety attacks in the past and laboratory tests were not performed .The
blood tests showed a glycemia of 35 mg/dl (70-110 mg/dl). She was admits for further evaluation. The laboratory
tests showed a c-peptid of 8.176 ng/ml (0.3-3.73 ng/ml) , insulin level of 35.66 uIU/ml (2.6-24.9 uIU/ml) and later
a proinsulin of 113 pmol/l (<11 pmol/l). After the test results came an abdominal computed tomography (CT) was
performed showing a singular mass in the tail of the pancreas .After the surgical removal of the mass blood glucose
levels reached normal levels without further hypoglycemic episodes.
CONCLUSIONS
Insulinomas in the beginning can be misdiagnosed or neglected so they can be difficult to diagnose and it needs
repeated evaluation and good communication between medical specialties and the patient.
Endocrinology
P1014
BROWN TUMOUR SECONDARY TO PRIMARY HYPERPARATHYROIDISM IN A YOUNG FEMALE
E. Madingwana 1, B. Chale-Matsau 1, M. Masipa 2, T. Pillay 1

1
Department of Chemical Pathology, University of Pretoria and National Health Laboratory Service, Tshwane Academic
Division, Pretoria, South Africa
2
Division of Endocrinology, Department Of Internal Medicine, Health Sciences , University of Pretoria, Pretoria, South Africa
BACKGROUND-AIM
The most common clinical presentation of primary hyperparathyroidism (PHPT) is asymptomatic
hypercalcaemia associated with an elevated or high-normal intact parathyroid hormone concentration. Atypical
presentations include several abnormalities in calcium homeostasis, ranging from normocalcaemic PHPT to severe
symptomatic hypercalcemia (parathyroid crisis). Parathyroid crisis is a rare manifestation characterized by high
calcium levels and marked symptoms of hypercalcaemia involving multiple organ systems. Extreme cases of PHPT may
present with metabolic encephalopathy, renal insufficiency or gastrointestinal symptoms.
METHODS
A 20-year-old female admitted to a general surgical ward at a tertiary hospital, presented with
chronic history of bone pain, significant weight loss and severe acute abdominal pain. She is not known with prior
medical conditions and no family history of malignancy
RESULTS
Biochemistry pattern revealed marked elevation of both serum calcium 4.3 mmol/L (2.15 – 2.50) and parathyroid
hormone > 265 pmol/L (1.6 – 7.2) associated with low phosphate 0.59 mmol/L (0.78 – 1.42). Alkaline phosphatase
was also increased 2359 IU/L (20 – 140). Vitamin D and renal function were within normal limits. Plain radiographs
of the pelvis and long bones showed lytic bone lesions, which were confirmed by computerised tomography scan
showed extensive generalised lytic bone lesions with the largest showing necrotic centred soft tissues components.
There were also T5 Lesions with vertebral plane, extra dural extension and kyphosis. Imaging of the kidneys revealed
nephrocalcinosis. Bone histology confirmed osteitis fibrosa cystica (brown tumour). The patient’s clinical, biochemistry
and radiological features confirmed severe primary hyperparathyroidism.
CONCLUSIONS
In patients presenting with hypercalcemia, regardless of age, primary hyperparathyroidism should be
considered a differential diagnosis. As observed in this patient, untreated primary hyperparathyroidism can result in
severe manifestations. Surgical treatment by parathyroidectomy can result in swift correction of serum calcium and
parathyroid hormone achieving a favourable prognosis.
Endocrinology
P1015
EVALUATION OF THYROID FUNCTION DURING FIRST TRIMESTER OF PREGNANCY
A. Atanasova Boshku 2, S. Kuzmanovska 1, E. Gjorgievska Nikolovska 2, I. Samardziski 2, V. Jovanovska 2

1
Institute of Pathophysiology and Nuclear Medicine, Faculty of Medicine, Ss Cyril and Methodius University
2
University Clinic of Gynecology and Obstetrics, Faculty of Medicine, Ss. Cyril and Methodius University Skopje, Skopje, North
Macedonia
BACKGROUND-AIM
Thyroid disorders could complicate pregnancy and pregnancy outcome, that’s why they ask for proper management of
this condition to avoid adverse maternal and fetal complications. Thyroid hormones are increasing in the first trimester
of pregnancy and their levels become stable in beginning of the second trimester. The level of thyroid hormones is very
important for fetal brain development. This study aims to assess the thyroid function in first trimester of pregnancy
in women with and without iodine supplementation to avoid potential maternal and fetal morbidity related to thyroid
dysfunction.
METHODS
In this study we have evaluate 181 pregnant women during the first trimester of pregnancy. From the study are
excluded pregnant women with preexistent diagnosed disorders, multifetal pregnancies and women with hypertensive
disorders. The patients were divided in two groups, first group of 89 women with normal pregnancy in first
trimester without iodine supplementation, and second group of 91 pregnant women in first trimester with iodine
supplementation (supplements with concentration of iodine of 150 µgr).
RESULTS
The analyzed results from group without iodine supplementation showed that the average level of TSH is 1.757 mIU/
L, and the group of women with iodine supplementation is 1.939 mIU/L. The average level of FT4 in women without
supplementation is 11.021 pmol/L, and in the group with iodine supplementation is 10.858 pmol/L. The average level
of TG in first group is 13.525 µgr/L, and in the second group with supplementation is 12.875 µgr/L.
CONCLUSIONS
Comparing the results of the average values of analyzed biomarkers of the thyroid gland, we concluded that there
is difference of the value of TSH with higher concentration in the examined group with iodine supplementation, and
difference in concentration of thyroglobulin with higher values in the group without iodine supplementation. The
results for all evaluated thyroid biomarkers are in the average range for all markers separately. Pregnant women living
in our country have sufficient supplementation with iodine intake, during the first trimester of pregnancy. But also,
depending of the thyroid gland function, individually are taken examinations and is included individual management
and treatment.
Endocrinology
P1016
CUSHING’S PATTERN IN URINE BY STEROID PROFILE.
E. Kowalska 1, A. Siejka 1, J. Klimiuk-Balas 1

1
The Children’s Memorial Health Institute
BACKGROUND-AIM
Cushing syndrome, which endogenous cause is in 80 % ACTH dependent, is related to ACTH secreting tumors or CRH
secreting tumors. In 20 % it can be a result of ACTH independent origin like adrenal tumors, carcinoma or macronodular
hyperplasia. Untreated hypercortisolemia causes significant morbidity and increase mortality due to its metabolic
effects including hypertension, obesity, osteoporosis, dyslipidemia and glucose intolerance. However, quick diagnose
is challenging. In Cushing’s disease not always all of the characteristics appear from the beginning. When alarming
disease features are observed a biochemical endocrine panel should be done, especially steroid profile in 24 hours
urine.
METHODS
Three patients in age 12-16 years (BMI 22-29) were diagnosed with Cushing’s disease. Steroid profile was performed
on a 24-hour urine collection by a gas chromatography (GC) coupled with 5973 mass selective detector (MSD) equipped
with an HP-ultra column 12 m. The MSD was operated in a single ion monitoring (SIM) mode.
RESULTS
Characteristic steroid metabolites were recognized in the urine. All of the patients with hypercortisolemia had evidently
high secretion of cortisol precursor metabolite which is tetrahydro-11-deoxycortisol (THS) with its concentration
exceeding from 4 to 17 times the highest normal level range. Other cortisol precursor metabolites were also elevated:
tetrahydrocortisone (THE), tetrahydrocortisol (THF), cortolons and cortols. Free cortisol (F) in 24 hours urine was
significantly elevated (from 13 to 41 times the highest normal level range) as well as free cortisone (E - from 3 to 14
times the highest normal level range). Also 5α-reductase deficiency was observed due to metabolites ratio.
CONCLUSIONS
Urinary steroid profile refers to many cortisol metabolites and its precursors in only one analysis, making it a good
diagnostic approach for hypercortisolemia suspicion. It is characterized by a high discriminatory value, especially in
a relation to the results obtained in obese patients. It is a non-invasive method that does not require hospitalization
of the patient.
Endocrinology
P1017
REFERENCE INTERVALS FOR HAIR CORTISOL IN HEALTHY ADULTS BASED ON ELECTROCHEMILUMINESCENCE METHOD
A. Tsitsimeli 2, A. Vergi 2, C. Kanaka-Gantenbein 2, A. Markou 1, A. Mantzou 2

1
Department of Chemistry, Lab of Analytical Chemistry, National and Kapodistrian University of Athens
2
Division of Clinical and Translational Research in Endocrinology, First Department of Pediatrics, School of Medicine,
National and Kapodistrian University of Athens, “Aghia Sophia” Children’s Hospital, 11527 Athens, Greece
BACKGROUND-AIM
Human scalp hair is a valuable matrix for determining long-term cortisol concentrations, with wide-spread applicability
in clinical care as well as research. In adults, population-based reference ranges for hair cortisol measurement have
been evaluated in healthy volunteers with liquid chromatography mass spectrometry. The aim of this study is to
establish age-adjusted reference intervals for hair cortisol in adults by using an automated immunoenzyme method
and determine the influence of gender and age.
METHODS
A total of 129 healthy adults were enrolled in the present study and their general characteristics (age, gender) were
collected. Scalp hair cortisol levels were measured using Cobas e 411 which is a fully automated analyzer that uses
a patented ElectroChemiLuminescence (ECL) technology for immunoassay analysis. Age-adjusted reference intervals
were established in adults from 18 to 80 years old.
RESULTS
Firstly, data were analyzed from two groups of healthy adults based on gender. Hair cortisol levels were statistically
significant higher in men than women (11,99pg/mg vs 6,57 pg/mg). Τhe cortisol levels of women were stable between
the different decades from 20 to 80 years whereas in men the cortisol levels were significantly elevated after the fifth
decade of life (23pg/mg vs 34,59 pg/mg, p<0.001). The variation of cortisol levels was smaller in women than in men.
CONCLUSIONS
This study provides knowledge about adult reference intervals and the effect of age and gender in cortisol levels, which
is essential for the implementation of hair cortisol testing and interpretation of the results in clinical practice however
a large number of healthy volunteer need to be analyzed.
Endocrinology
P1018
EFFECT OF NOCTURNAL MELATONIN INTAKE ON SLEEP QUALITY ACCORDING TO THE MENSTRUAL CYCLE
S. Fendri 2, R. Makhlouf 2, A. Elleuch 2, I. Kamel 1, O. Hammouda 1, F. Ayadi 2, M. Naifar 2

1
Higher Institute of Sport and Physical Education of Sfax
2
Laboratory of Biochemistry, CHU Habib Bourguiba
BACKGROUND-AIM
Melatonin is the main hormone for regulating the nychthemeron and the sleep-wake cycle. We attempt through this
work to research the effects of nighttime intake of melatonin in women on the quality of sleep according to the
menstrual cycle.
METHODS
Ten healthy young women between the ages of 20 and 23 were included in our study after having had their consent.
Various anthropometric parameters were collected (weight, height, BMI, percentage of fat). The experimental protocol
consists of a randomized nocturnal intake of placebo and melatonin around 9 p.m. followed by a set of questionnaires
the next morning around 9 a.m. concerning sleep (Hopper sleep and fatigue, POMS, spiegel, vis Morgan and DLQ). This
is done once in the middle of the follicular phase and once in the middle of the luteal phase.
RESULTS
The average age of the participants is 21.63 ± 0.94 years. The average height of our patients is 165.72 ± 3.38 m. the
average weight of our patients is 59.45 ± 3.5 kg.
For the sleep and fatigue Hopper score and the POMS and DLQ score, there was no significant difference between the
follicular phase and the luteal phase or between taking melatonin or placebo.
For the Spiegel score, the sleep score is significantly higher during the luteal phase compared to the follicular phase
(p<0.01). The sleep score is significantly higher after taking melatonin compared to that of placebo during the follicular
phase (p=0.027) and the luteal phase (p=0.03).
For the Morgan score, the sleep score is significantly higher during the luteal phase compared to the follicular phase
(p=0.03). The sleep score is significantly higher after taking melatonin compared to that of placebo during the follicular
phase (p<0.01).
CONCLUSIONS
The effects of melatonin on sleep according to the menstrual cycle are variable according to the scores used and in
all cases beneficial.
Endocrinology
P1019
TSH REFERENCE INTERVALS IN THE FIRST YEAR OF LIFE – HOW MANY AGE PARTITIONS SHOULD WE USE?
U.D. Senarathne 1, Z.X. Lu 1, J.C.G. Doery 2, A. Mcneil 5, N. Wijeratne 5, D. Gangoda Liyanage 3, J. Smith 4, E.M. Lim 8, P.
Coates 9, M. Saleem 9, N. Heather 6, C. Kyle 6, R.C. Heron 6, M. Zacharin 7, J. Brown 3, J.K. Hewitt 3
1
Department of Biochemistry, Monash Health Pathology, Monash Health, Victoria, Australia
2
Department of Medicine, Monash University, Victoria, Australia
3
Department of Paediatric Endocrinology, Monash Children’s Hospital, Victoria, Australia
4
Department of Pathology, Royal Children’s Hospital, Victoria, Australia
5
Dorevitch Pathology, Victoria, Australia
6
Lab Plus, Te Toka Tumai, Auckland, New Zealand
7
Murdoch Children's Research Institute, The Royal Children’s Hospital, Victoria, Australia
8
Path West Laboratory Medicine QEII, Department of Endocrinology and Diabetes, Sir Gairdner Hospital, West Australia,
Australia
9
SA Pathology, South Australia, Australia
BACKGROUND-AIM
There is no consensus on what age partitions should be used for defining Thyrotropin (TSH) reference intervals (RI)
in the first year of life. Correct diagnosis and treatment of hypothyroidism in an infant require age-appropriate RI for
interpretation. We describe six age groups based on the significant changes in TSH values in apparently healthy infants.
METHODS
De-identified TSH data (Beckman DxI800) from 2005-2021 were extracted from the Monash Health Pathology
database. Exclusions were: (1) more than one episode of testing, (2) known thyroid disease, TSH <0.01 or >50 mIU/L, or
(3) conditions that could affect thyroid function, including prematurity, severe illnesses, or presence of positive thyroid
antibodies. Age partitions were defined by visual inspection of the scatter plot of TSH against age, and differences in
TSH concentrations between the proposed age partitions were confirmed using the Mann-Whitney U test.
RESULTS
Of 13,245 episodes of TSH testing in infants, 5,876, 1,811, and 149 episodes, respectively, were excluded for each step
listed above. The remaining 5,409 infants with only one episode of TSH were used for defining age partitions.
On visual examination of the TSH plot against age, TSH decreased more rapidly in the first two weeks of age and then
gradually up to 90 days, but no obvious change was observed after 90 days. Mann-Whitney U test performed on daily
TSH sequentially from days 0-13 showed that TSH was significantly changed during the first four days of life (days-0&1
vs. day-2: p<0.001; day-2 vs. day-3: p<0.001; day3 vs. day-4: p=0.018) but not different from day-4 to day-13 (p-value
ranged from 0.22 to 0.94). No significant difference in weekly TSH was observed from days 14-89 (p-values ranged from
0.14 to 1.0). When the Mann-Whitney U test was performed on monthly TSH values from day-60 onwards, a significant
difference between month-3 (days 60-89) and month-4 (days 90-119) was found (p=0.0013), but no differences in
monthly TSH from day 90 of life onwards (p-value ranged 0.54 to 0.96).
CONCLUSIONS
The age partitions were then defined based on these TSH patterns representing dynamic physiological changes in
thyroid function in the first year of life: (1) age 0&1 days; (2) age 2 days; (3) age 3 days; (4) age 4-13 days; (5) age 14-89
days; and (6) age 90-365 days.
Endocrinology
P1020
AGE-SPECIFIC TSH REFERENCE INTERVALS PARTITIONED BASED ON THE PHYSIOLOGICAL CHANGE IN THYROID
FUNCTION IN THE FIRST YEAR OF LIFE
Z.X. Lu 1, U.D. Senarathne 1, D. Gangoda Liyanage 3, J.C.G. Doery 2, J. Smith 4, A. Mcneil 5, N. Wijeratne 5, E.M. Lim 8, N.
Heather 6, C. Kyle 6, R.C. Heron 6, P. Coates 9, M. Saleem 9, M. Zacharin 7, J. Brown 3, J.K. Hewitt 3
1
Department of Biochemistry, Monash Health Pathology, Monash Health, Victoria, Australia
2
Department of Medicine, Monash University, Victoria, Australia
3
Department of Paediatric Endocrinology, Monash Children’s Hospital, Victoria, Australia
4
Department of Pathology, Royal Children’s Hospital, Victoria, Australia
5
Dorevitch Pathology, Victoria, Australia
6
Lab Plus, Te Toka Tumai, Auckland, New Zealand
7
Murdoch Children's Research Institute, The Royal Children’s Hospital, Victoria, Australia
8
Path West Laboratory Medicine QEII, Department of Endocrinology and Diabetes, Sir Gairdner Hospital, West Australia,
Australia
9
SA Pathology, South Australia, Australia
BACKGROUND-AIM
Hypothyroidism in infancy is often clinically asymptomatic and can only be confirmed by biochemical testing. The
dynamic physiological changes in thyroid function during this period necessitate the use of age-specific thyrotropin
(TSH) reference intervals (RI) to avoid delayed treatment. Previously published TSH RI studies were limited by study
sample sizes and inappropriate age-partitioning, especially in the first month of life. In the absence of international
consensus on age partitions and the respective TSH RI for infants, we have recently defined appropriate age
partitioning into six groups reflecting physiological changes in TSH in infancy (data presented in an accompanying
abstract). We aimed to establish definitive TSH RI based on these age partitions for the common analytical methods
used in Australia by an indirect approach using data mining. We hereby present TSH RI for the Beckman DxI, Siemens
Centaur and Atellica, and Abbott Architect methods.
METHODS
De-identified TSH data from infants were extracted from multiple pathology laboratories. After excluding TSH results
from infants with more than one TSH measurement, TSH <0.01 or >50 mIL/L, thyroid disease or severe illnesses, or
premature birth, TSH results from 22,931 infants were included in the analysis. RI was determined using the 2.5th and
97.5th percentiles of TSH values in each partition.
RESULTS
The TSH RI (n) for the age partitions of age 0 & 1 days, 2 days, 3 days, 4-13 days, 14-89 days, and 90-365 days were:
2.6-39.8 (122), 1.8-14.9 (172, 1.1-10.3 (242), 1.2-8.5 (2476), 1.1-7.8 (1398) and 0.6-5.7 (999) mIU/L respectively for
the Beckman method; 1.5-38.9 (288), 1.1-15.3 (352), 1.0-13.1 (453), 0.9-8.5 (4638), 0.8-8.1 (3613) and 0.6-6.6 (3740)
mIU/L respectively for the Siemens method; and 0.8-39.0 (102), 1.1-16.5 (142), 0.7-11.2 (195), 0.6-8.2 (1178), 0.9-7.0
(1146) and 0.6-4.6 (1675) mIU/L respectively for the Abbott method. No difference was observed between male and
female infants.
CONCLUSIONS
To our knowledge, this is the first study addressing TSH RI in infancy for multiple analytical methods with a
standardised approach and by using statistical methods to appropriately define the age partitions with the respective
TSH RI reflecting physiological changes in thyroid function in infants.
Endocrinology
P1021
REFERENCE INTERVALS OF THYROID STIMULATING HORMONE, THYROID HORMONES, AND THYROID
AUTOANTIBODIES IN ADULT AND ELDERLY THAI INDIVIDUALS.
S. Tanticharoenkarn 2, P. Pipatnavakij 2, L. Piyasuwanying 2, P. Ganokroj 1

1
Department of Laboratory Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
2
Department of Laboratory Medicine, King Chulalongkorn Memorial Hospital, The Thai Red Cross Society, Bangkok, Thailand
BACKGROUND-AIM
Reference intervals (RIs) for thyroid function tests and thyroid autoantibodies are population-, age-, and assay-specific.
This study aimed to establish RIs for Thai adults and elderly individuals.
METHODS
A cross-sectional study was conducted at a single tertiary center from October 2021 – October 2022. Recruited
individuals were divided into two groups, adults (aged 18 – 59) and elderly (aged ≥60). The screening method
consisted of 1. An interview of conditions/medications that affected thyroid status, 2. A thyroid ultrasonography to
exclude abnormal thyroid structures, and 3. Laboratory tests to include only individuals with negative for thyroid
autoantibodies: thyrotropin receptor antibody (TRAb) by electrochemiluminescence immunoassay, thyroglobulin
antibody (TgAb), and thyroid peroxidase antibody (TPOAb) by chemiluminescence immunoassay (CMIA). The analyses
for TSH, FT4, TT4, FT3, and TT3, were performed by the Abbott Alinity I analyzer with the CMIA method. The studies
for 2.5th - 97.5th values were committed to establishing RIs.
RESULTS
A total of 356 individuals were screened, and 216 (112 adults, 54% women, and 104 elderly, 50% women) were included
in the analysis. The median age and BMI in the adult group VS the elderly group were 33.5 years and 22.1 kg/m2 VS 65.5
years and 23.8 kg/m2, respectively. The RIs of TSH was 0.39 – 4.17 mIU/L in all, 0.35 – 3.98 mIU/L in the adult group,
and 0.42 – 4.83 mIU/L in the elderly group. The RIs of FT4, TT4, FT3, and TT3 were 0.75 – 1.09 ng/dL, 4.46 – 9.38 ng/dL,
2.10 – 3.27 pg/mL, 58.72 – 115.45 pg/mL in all, 0.76 – 1.06 ng/dL, 4.23 – 9.49 ng/dL, 2.15 – 3.36 pg/mL, 60.45 – 118.44
pg/mL in the adult group and 0.74 – 1.15 ng/dL, 4.23 – 9.46 ng/dL, 1.99 – 2.94 pg/mL, 52.14 – 108.88 pg/mL in the
elderly group, respectively. According to this study's newly established RIs of thyroid autoantibodies, the prevalences
of positive titer for TgAb and TPOAb were 21.3% and 14% in adults VS 27.3% and 24% in the elderly, respectively.
CONCLUSIONS
In the elderly group, TSH levels increase, FT3/TT3 levels tend to be lower, and FT4/TT4 levels are comparable to adults.
The age-specific TSH level should be considered in evaluating thyroid disorders in aging.
Endocrinology
P1022
LABORATORY INTERVENTION IN A CASE OF MACROAMYLASEMIA
A. Delgado Baena 1, J.M. Villa Suárez 1, P. Parrado Hernando 1, C. Martín Tovar 1, L. Velasco Rodríguez 1, J.M. Molina
Santiago 1
1
Laboratory/ Hospital San Agustín, Linares
BACKGROUND-AIM
The presence of increased serum amylase usually implies the existence of a pancreatic or parotid disease. However,
there are situations in which hyperamylasemia is not related to increased secretion of the enzyme or to renal failure.
METHODS
A 29-year-old woman who underwent a routine check-up by Primary Care because she presented dispepsia. The
laboratory report revealed the following data: α-amylase 1142 U/L (Reference value: 28-100 U/L), lipase 30 U/L,
aspartate transaminase 36 U/L, alanine transaminase 31 U/L.
After that, she is referred to the Digestive Service, which requests a second sample accompanied by some
complementary tests for the study of hyperamylasemia: Biochemistry; α-amylase 1014 U/L, lipase 28 U/L, aspartate
transaminase 9 U/L, alanine transaminase 12 U/L. Urine; α-amylase 119 U/L (Reference value: 0 – 460 U/L).
Immunology; Anti-transglutaminase antibodies negative. Tumor markers; carcinoembryonic antigen and CA 19.9, both
normal. Serology; Negative hepatitis and human immunodeficiency virus.
After these findings, the following differential diagnoses are ruled out: acute pancreatitis, viral origin, tumor origin
and celiac disease. For this reason, a possible case of macroamylasemia begins to be suspected. Given the capacity
of polyethylene glycol (PEG) 6000 for the differential precipitation of macromolecules, the monomeric fraction in the
supernatant resulting from the following test was measured:
- First tube: 200 µL of serum and 200 µL of 25% PEG solution were mixed.
- Second tube: 200 µL of serum and 200 µL of distilled water were mixed.
RESULTS
The concentration of α-amylase in the supernatant was analyzed. Tube 1 result: 44.83 U/L; tube 2 result: 509.81 U/L.
The percentage of precipitated α-amylase is obtained: ((Tube 2 – tube 1)/tube 2) x 100. Result: 91.2%. A percentage of
precipitation greater than or equal to 73% implies the existence of macroamylasemia, thus confirming the diagnosis.
CONCLUSIONS
In some diseases, abnormal protein synthesis occurs, with the potential to bind to α-amylase and produce a complex
with enzyme activity. Macroamylasemia should always be considered before any patient who presents atypical clinical
characteristics of pancreatitis, especially if the amylase elevation persists over time and the symptoms have subsided.
Endocrinology
P1023
THYROGLOBULIN MEASUREMENT: PERFORMANCE VERIFICATION OF THE BRAHMS KRYPTOR HTG ASSAY AND
COMPARISON WITH A RADIOIMMUNOASSAY.
L. Courcelles 1, L. Wauthier 1, D. Gruson 1

1
Cliniques Universitaires Saint-Luc
BACKGROUND-AIM
Thyroglobulin (Tg) measurement is a specific and sensitive biomarker for the monitoring of thyroid cancer after total
thyroidectomy. The presence of Tg may be suggestive of cancer recurrence or improper surgery. Despite their clinical
relevance, immunoassays suffer from interference with anti-Tg antibodies occurring in about 20% of thyroid cancer
patients. The antibodies bind with free Tg and may lead to false-negative results. Therefore, radioimmunoassays (RIA)
have been used as alternative methods in presence of anti-Tg. We decided to evaluate the on-site performance of hTg
Kyptor assay and compare it with RIA in samples positive for anti-Tg.
METHODS
According to CLSI guidelines, we determined our internal reference values on 20 healthy volunteers and evaluated
imprecision with two levels of quality control (QC). For the method comparison, we analyzed 33 samples of patients with
thyroid disease with both the RIA and the Brahms hTG Kryptor assay. Method comparison between the two methods
was performed using Passing-Bablok regression and Bland Altman plots. We also calculated inter-rater agreement
(kappa) at the limit of quantification (LOQ) (0.5 ng/ml for the kryptor and 0.2 ng/ml for the RIA).
RESULTS
For the Kryptor assay, our local reference interval was 7.28 – 56.79 ng/ml (95% confidence interval (CI)) versus 0.3 - 58
ng/mL in the insert kit. The intraassay and interassay coefficient variations were respectively 2.7% and 3.4% for the low
QC level and 2.2% and 2.4% for the high QC level. Method comparison showed a regression equation slope of 1.40 (95%
CI: 1.36–1.56) and an intercept of -0.08 ng/ml (95% CI: -0.11– -0.02). The mean difference between methods was 29.6%
(95% CI: 18.6–40.7). The concordance analysis demonstrated a Kappa coefficient of 0.88 (95% CI :0.71-1) at the LOQ .
CONCLUSIONS
We adjusted our local reference values for the Kryptor hTg assay as it varied from the manufacturer. In terms of
precision, our results matched their analytical goals. Comparing to the RIA, the two methods are not commutable
although there is a very good agreement between the two at the LOQ. Clinical decisions should be adjusted accordingly.
In the presence of anti-Tg, this method may benefit to clinical laboratories as it is seems less susceptible to
interferences, time saving and fully automated.
Endocrinology
P1024
EVALUATION OF VITAMIN D (25 OH) MEASUREMENTS IN 2022, CONCLUSIONS AND CURRENT GUIDELINES
A. Momirovska 1, M. Todorovska 1, A. Georgieva 1, A. Georgievska 1

1
PHI SYNLAB Macedonia
BACKGROUND-AIM
Vitamin D is no longer important solely for prevention and cure of rickets, mineral metabolism and musculoskeletal
health. It is related with extra-skeletal diseases such as cancer, cardiovascular diseases, immunity and infections. Low
Vitamin D level is indicator of poor health status.
Evaluation of low prevalence of Vitamin D levels is important for public health perspective. In that sense we evaluated
our results from 2022 and offer them for further analysis. Also we present dosage recommendations in accordance to
the guidelines of Synlab Academy for optimal dosage of Vitamin D.
METHODS
380 Serum samples were measured for Vitamin D total (25 OH) using ECLIA/ ROCHE cobas e 411. Samples were collected
from three locations in Skopje, one location in Tetovo and one location in Bitola. Results were analyzed regarding
reference values, location of sampling and gender of the patients.
RESULTS
156 (41.06 %) patients had results lower than 20 g/L (Vit D deficiency). 102 (26.95 %) examinees had level between
20-30 g/L (Vit D insufficiency). 117 (31 %) patients were in range 30-100 g/L (therapeutic levels) and 0.94 % patients
had intoxication levels over 150 g/L.
In the patients from Tetovo was found higher level 59.25 % of deficiency, versus Skopje with about 37 % Vit D deficiency
and Bitola with 28.55 %.
Therapeutic levels (30-100 g/L) were found in 36 % patients from Skopje and 16.6 % patients in Tetovo. Bitola
percentage is almost same as in Skopje.
68.2 % examinees were females and deficiency was found in 62% of them.
CONCLUSIONS
Our results show that in our country deficiency of Vitamin D is present with high percentage. Results are alarming
for population in Tetovo 59.25%, especially in females. That can be result of cultural behavior of Muslim population
in Tetovo and non-exposure to daylight. Also high percentage of lactose intolerance in our country (about 70%)
contributes for low intake of Vitamin D from food.
Highest therapeutic range is find in Skopje, the capitol, which means that part of population in Skopje takes preventive
nutritional, behavioral and supplemental measures.
Recommendation for all patients with deficiency and insufficiency (400-800 IU Vit D per day) should be included in all
lab reports signed by medical lab specialists to improve general Vitamin D perspective.
Endocrinology
P1025
FIRST TRIMESTER REFERENCE RANGE FOR THYROID FUNCTION TESTING AMONG PREGNANT WOMEN SCREENED AT A
TERTIARY CARE HOSPITAL IN R.N. MACEDONIA
A. Atanasova Boshku 2, V. Jovanovska 2, S. Kuzmanovska 1, A. Shabani 2

1
Institute of Pathophysiology and Nuclear Medicine, Faculty of Medicine, Ss Cyril and Methodius University
2
University Clinic of Gynecology and Obstetrics, Faculty of Medicine, Ss. Cyril and Methodius University Skopje, Skopje, North
Macedonia
BACKGROUND-AIM
Maternal thyroid dysfunction has been associated with adverse pregnancy outcomes. The purpose of our study was to
evaluate and establish first trimester specific reference intervals for thyroid hormones in pregnant women in tertiary
care hospital setting.
METHODS
One hundred eighty women with less than 14 weeks gestation were screened at a tertiary care hospital in Skopje,
Macedonia. Serum levels of thyroid-stimulating hormone (TSH), free thyroxine (fT4), thyroglobulin and thyroid
autoantibodies were measured using Siemens Immulite 2000 HP.
RESULTS
GRIs (2.5th and 97.5th percentiles) were determined for TSH and fT4. Effect of maternal age, gestational age, and
maternal weight was analyzed. Statistical Analysis Quantitative variables were expressed as means and standard
deviations (SD), and qualitative variables were expressed as frequencies and percentages. Normality of the data was
checked using the Kolmogorov–Smirnov test. Values that were normally distributed were expressed only as means and
SD. Those that were not normally distributed were expressed as medians and interquartile range. For all statistical
analysis, p < 0.05 was considered as statistically significant. Results First-trimester GRI was 0.245 to 4.971 mIU/L for
TSH, 10.2 to 18.9 pmol/L for fT4.
CONCLUSIONS
These laboratory and first trimester specific GRI for TSH and fT4 shall help in proper diagnosis and treatment of
subclinical thyroid dysfunctions.
Endocrinology
P1026
ANTI-MULLERIAN HORMONE AS OVARIAN RESERVE SERUM BIOMARKER IN OUR POPULATION
N. Padilla Apuntate 2, C.G. Puerto Cabeza 2, C. García Carro 2, N. Goñi Ros 2, M.Á. Castillo Arce 2, M.Á. César Márquez 1
1
University Hospital Miguel Servet, Zaragoza, Spain
2
University Hospital Miguel Servet, Zaragoza, Spain.
BACKGROUND-AIM
The term ovarian reserve (OR) has traditionally been used to describe women’s reproductive potential. It was used to
be evaluated by follicle-stimulating hormone (FSH), which is produced and released by the gonadotropic cells of the
adenohypophysis. Recently, FSH has been replaced by Anti-Müllerian hormone (AMH). AMH is secreted by the ovarian
granulosa cells of follicles. The aim of this study is to evaluate the utility of AMH as a serum biomarker of OR in our
population.
METHODS
Study target group: 78 women (Mean Age: 35,63,16 ages) undergoing assisted reproduction during January and
November 2020.
We conducted a retrospective descriptive statistical study to confirm the relationship between AMH levels and antral
follicles count (AFC) observed into the right and left ovaries by transvaginal ultrasound in each patient, that constitute
OR.
FSH was measured by paramagnetic particle chemiluminescence immunoassay in DxI 800 (Beckman Coulter®) and
AMH was measured by electrochemiluminescence immunoassay in Cobas e411 (Roche®), both in serum samples.
The database and statistical data analysis were performed with Microsoft Excel 2010. The study population was divided
into two groups: (1) low OR when AFC<7 and (2) high OR when AFC≥7. The relationship between the variables AMH and
OR, and FSH and OR was illustrated by a box plot. AMH and FSH means of each group were compared with a T-test.
RESULTS
Those women who had low OR presented low AMH levels (x=0,56ng/mL) and high FSH levels (x=9,85mUI/mL) while
women with high AMH levels (x=2,48ng/mL) and low FSH levels (x=7,9mUI/mL) presented high OR.
A clearer distinction was observed between AMH means than FSH means in the box plot. AMH means in the high group
with respect to the low group presented a lower p value (p<0,001) than FSH means (p=0,02) for the T-test.
CONCLUSIONS
Low AMH levels are associated with fewer AFC, which can help diagnose infertility in women or indicate that there is a
lower chance of becoming pregnant. FSH levels and AFC have been related, resulting in an inverse relationship between
them.
This study demonstrates the close relationship between OR and AHM levels, in our population. Moreover, as FSH
normally varies during ovarian cycle and AMH remains stable, that justify its utility as an ovarian reserve serum
biomarker.
Endocrinology
P1027
DIFFERENTIATION OF SIMPLE OBESITY IN THE DIAGNOSIS OF STEROIDOGENESIS DISORDERS.
J.T. Stradomska 1, A. Siejka 1, E. Kowalska 1, J. Klimiuk-Balas 1, E. Moszczyńska 1

1
The Children’s Memorial Health Institute
BACKGROUND-AIM
Obesity is one of the basic clinical symptoms in a number of steroidogenesis disorders. On the other hand, simple
obesity is a chronic disease that can lead to a development of the other lifestyle diseases. Currently overweight and
obesity are increasing in society, not only in adults, but also in pediatric population.
The aim of the study was to develop a pattern of steroid hormones in the urinary steroids profile to allow a
differentiation of simple obesity from the disorders of steroidogenesis process.
METHODS
In the years 2021-2022, 575 patients (397 girls and 178 boys) aged 4-17 years old were referred to the Laboratory
of Steroid Hormones and Metabolic Disorders of the CMHI in order to analyze steroid profile for suspicion of
steroidogenesis disorders. In the group under investigation: 132 patients were suspected of hypercortisolemia;
207 with precocious puberty or suspected of congenital adrenal hyperplasia (CAH) or a non-classical form of
congenital adrenal hyperplasia (NC-CAH); 236 with hypothyroidism, menstrual disorders, and insulin resistance. All
subjects presented overweight or obesity. All patients underwent an analysis of the urinary steroids profile by mass
spectrometry - gas chromatography method (GC-MS).
RESULTS
In the study group, Cushing's syndrome was confirmed in 4, NC-CAH in 19, and hypothyroidism in 153 patients,
376 presented premature adrenarche. Whereas 23 patients presented only simple obesity with no steroidogenesis
disorders. The steroids profile of the patients with simple obesity showed an over 3-fold concentration increase of
corticosteroids in relation to reference values without signs of cortisol metabolism disorders. A significant increase of
androsterone/etiocholanolan ratio indicates an increase of 5α-reductase activity in comparison to the other previously
mentioned disorders.
CONCLUSIONS
The characteristic pattern of steroids in the analyzed urinary steroids profile of patients with simple obesity allows
them to be excluded from the group suspected of a defect in the steroidogenesis process.
Endocrinology
P1028
DETERMINATION OF URINARY STEROIDS IN HEALTHY WOMEN BY GC-MS METHOD
A. Siejka 1, T.J. Stradomska 1, M. Jaworski 1, A. Calik 2

1
Dpt Biochemistry, Radioimmunology & Experimental Medicine, The Children’s Memorial Health Institute
2
Dpt Gynecological Endocrinology, Medical Uniwersity of Warsaw
BACKGROUND-AIM
Radioimmunological methods used for the determination of steroid hormones and their metabolites do not meet the
required standards for analytical procedures, due to known limitations in immunological processes. An alternative
analytical technique is the determination of the urinary steroid profile using gas chromatography-mass spectrometry
(GC-MS) methods. The reliability of diagnostics results is based on a precisely validated method and reliable
determination of reference values.
Objective
Determination of the reference values of individual steroid hormones in the steroids profile covering 40 metabolites
in women of reproductive age.
METHODS
Subjects:
- 150 females, aged 16 – 40 years without signs of gynecological and endocrine disorders,
Exclusion criterion: taking hormonal drugs, pregnancy, inappropriate phase of the cycle.
- 5 patients manifested insulin resistance, menstrual disorders, hirsutism, hyperandrogenism
- One patient with 11-βhydroxylase deficiency – monitoring of encortone therapy.
Study material – 24 h-urine sample collection.
Biochemical analysis:
Urinary steroids were measured by adaptation of the selected –ion – monitoring (SIM) gas chromatography - mass
spectrometry GC/MS method.
RESULTS
Reference values have been established for 40 steroids. Data were analyzed using Statistica PL, version 10. Normality
of the distribution of variables was tested by the Shapiro-Wilk W test. Non-parametric statistics were used. Median
and percentiles: 2.28 and 97.72 were calculated as non-parametric equivalents of mean and +/- 2 standard deviation.
Elevated 17-OHP and androgen metabolites were found in the group of 5 patients with insulin resistance, menstrual
disorders, hirsutism, and hyperandrogenism. It suggests observation towards PCOS. The analysis of steroid profiles,
conducted in a patient with 11-βhydroxylase deficiency during therapy, confirms the usefulness of the method for
monitoring and evaluating treatment results.
CONCLUSIONS
Determination of urinary steroids by the GC-MS method combined with the analysis of steroid synthesis and
metabolism pathways makes it possible to isolate new pathologies and propose new pathomechanisms in endocrine
disorders. What is more, it can be used for a personalized approach to diagnosis and differential diagnosis.

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Poster Abstracts – WorldLab • EuroMedLab Roma 2023 – Rome, Italy, May 21-25, 2023 • DOI 10.

A. Al Hayek 1, S. Sobki 2, S. Sobki 2

S. Fendri 2, M. Boudaya 2, R. Ben Salah 1, K. Jamoussi 2, Z. Bahloul 1

L. Roli 1, S. Tagliavini 2, M.C. De Santis 2, S. De Vincentis 3

K. Hutchinson 2, H. Ali 2, M. Louw 2, J. Faul 1

J. Montenegro-Martínez 1, R. Sánchez-Rubio 1, E. Lepe-Balsalobre 1, J.M. Guerrero Montávez 1

M. Herrmann 1, S. Zelzer 1, M. Kleber 3, C. Drexler-Helmberg 2, P. Schlenke 2, P. Curcic 1, D. Enko 1, H. Scharnagl 1, S. Pilz

E. Dworniczak 4, E. Zapala 4, J. Brąszkiewicz 5, M. Szczur 5, M. Nowacka 3, C. Lafrence 1, A. Ruzhanskaya 2

F. Ponzetto 3, M. Parasiliti Caprino 3, F. Settanni 1, A. Nonnato 1, E. Ghigo 3, R. Giordano 2, G. Mengozzi 1

F. Ponzetto 2, F. Settanni 1, A. Nonnato 1, R. Nicoli 3, E. Ghigo 2, K. Tiia 3, G. Mengozzi 1

K. Siﬁ 3, S. Hanachi 3, K. Boudaoud 2, S. Zekri 3, K. Benmohamed 1, N. Nouri 1, K. Benembarek 3, N. Abadi 3

C. Le Goff 1, C. Defourny 3, J. Deleersnyder 2, G. Collard 2, N. Ferrante 2, S. Peeters 1, E. Cavalier 1

B. Netzel 1, A. Maus 1, S. Grebe 1

E. Grifnée 1, F. Blistein 3, J. Demeuse 2, L. Huyghebaert 1, P. Massonnet 1, M. Schoumacher 2, T. Dubrowski 1, S. Peeters

J. Demeuse 2, P. Massonnet 1, E. Grifnée 1, L. Huyghebaert 1, M. Schoumacher 1, T. Dubrowski 1, W. Determe 1, S. Peeters

J. Brąszkiewicz 5, M. Szczur 5, E. Dworniczak 4, E. Zapała 4, M. Nowacka 3, L. Cesili 1, A. Ruzhanskaya 2

L. Calton 1, D. Foley 1, G. Hammond 1, N. Breen 4, P. Rossiter 4, G. Bosch 2, S. Donzelli 3

C. Le Goff 2, S. Peeters 1, N. Vanden Broeck 1, A. Gendebien 1, C. Nix 1, M. Rechchad 1, E. Cavalier 2

M.P. Genova 2, B. Atanasova 2, I. Ivanova 3, K. Todorova 5, K. Tosheva 4, T. Daskalova 1

H. Van Rossum 1, L. Van Winden 1, A. Bergman 1, H. Van Der Poel 1

L. Garre Morata 1, C. Aldana GarzÓn 1, A. Perojil Jimenez 1, T. De Haro MuÑoz 1

P. Samarakoon 1, S. Gunawardena 1, T. Inthujah 1, G. Dissanayake 1, M. Balasooriya 1, S. Sayanthan 1, Z. Thaneefa 1, T.

N. Díaz-Troyano 1, M. Giralt-Arnaiz 1, C. Aguilar-Riera 2, D. Yeste-Fernández 2, R. Ferrer-Costa 1, M. Clemente-León 2

C. Le Goff 1, S. Peeters 2, M. Rechchad 2, J. Deleersnyder 2, E. Cavalier 1

B. Beljan 1, V. Šupak Smolčić 2, L. Bilić-Zulle 2

J. Rhea-Mcmanus 1, M. Quintanilla 1, H. Zhang 1, M. Guo 1, H. Leipold 1

J. Tortosa-Carreres 1, R. Andreu-Escriva 1, A. Alba-Redondo 1, M.L. Martínez-Triguero 1, B. Laiz-Marro 1

I. Perovic Blagojevic 6, J. Vekic 4, D. Macut 2, S. Ignjatovic 5, M. Miljkovic-Trailovic 3, A. Zeljkovic 3, I. Bozic-Antic 1, J.

T.U. Athapaththu 1, S. Peduruhewa 1

D. Casanova Expósito 1, C. Valldecabres Ortiz 1, S. Górriz Pintado 1

T.M.N.K. Puliyadda 1, G. Premawansa 2, B.K.T.P. Dayanath 1

T.M.N.K. Puliyadda 1, B.K.T.P. Dayanath 1

S. Marchiani 2, F. Villanelli 2, S. Dabizzi 1, S. Degl'Innocenti 1, M.G. Fino 1, L. Vignozzi 2, G. Danza 2

M. Bello 1, A. Ajose 1, K. Mosaku 2, T. Adedeji 1, A. Ajeigbe 1

E. Talbi 1, T. Mazouni 1, R. Ghodbane 1, L. Zerelli 1, S. Oueslati 1, S. Hammami 1, R. Mahjoub 1

A. Krol 2, K. Bornikowska 1, M. Ostrowska 1, P. Glinicki 1, W. Zgliczynski 1, K. Kowalski 2

A. Krol 2, K. Bornikowska 1, M. Ostrowska 1, P. Glinicki 1, W. Zgliczynski 1, K. Kowalski 2

M.C. Martín Fernández De Basoa 1, M.T. Concepción Masip 1

S. Tombari 1, Y. Amri 1, Y. Salem 2, Y. Hasni 2, R. Dabboubu 1, S. Ferchichi 3, t. Messaoud 1

M. Rogozinska 2, R. Rola 1, K. Kowalski 1, T. Bienkowski 1

V.I. Mayasari 2, A. Aryati 2, H.E. Sidjabat 3, B.P. Semedi 1, F.R. Marpaung 2

M. Taglieri 1, L. Di Gregorio 1, B. Donati Marello 1, A. Di Guida 1, P. Merlach 1, T. Manetta 1, G. Mengozzi 1

A. Oniye 1, M. Charles- Davies 1, O. Olaopa-Olapade 2, J. Anetor 1, O. Agbedana 1

D. Wittekind 5, K. Jürgen 5, M. Roland 3, K. Wirkner 4, R. Baber 5, C. Sander 2, V. Witte 1, A. Villringer 1, M. Kluge 2

M. Kurshumliu 1, I. Ibrahimi 2, A. Zeka 3

E. Madingwana 1, B. Chale-Matsau 1, M. Masipa 2, T. Pillay 1

A. Atanasova Boshku 2, S. Kuzmanovska 1, E. Gjorgievska Nikolovska 2, I. Samardziski 2, V. Jovanovska 2

E. Kowalska 1, A. Siejka 1, J. Klimiuk-Balas 1

A. Tsitsimeli 2, A. Vergi 2, C. Kanaka-Gantenbein 2, A. Markou 1, A. Mantzou 2

S. Fendri 2, R. Makhlouf 2, A. Elleuch 2, I. Kamel 1, O. Hammouda 1, F. Ayadi 2, M. Naifar 2

S. Tanticharoenkarn 2, P. Pipatnavakij 2, L. Piyasuwanying 2, P. Ganokroj 1

L. Courcelles 1, L. Wauthier 1, D. Gruson 1

A. Momirovska 1, M. Todorovska 1, A. Georgieva 1, A. Georgievska 1

A. Atanasova Boshku 2, V. Jovanovska 2, S. Kuzmanovska 1, A. Shabani 2

J.T. Stradomska 1, A. Siejka 1, E. Kowalska 1, J. Klimiuk-Balas 1, E. Moszczyńska 1

A. Siejka 1, T.J. Stradomska 1, M. Jaworski 1, A. Calik 2

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