Next-generation sequencing (NGS) followed by metagenomic enables the detection and identification of known as well as novel pathogens. It could be potentially useful in the diagnosis of encephalitis, caused by a variety of microorganisms.... more
Next-generation sequencing (NGS) followed by metagenomic enables the detection and identification of known as well as novel pathogens. It could be potentially useful in the diagnosis of encephalitis, caused by a variety of microorganisms. The aim of the present study was to evaluate the sensitivity of isothermal RNA amplification (Ribo-SPIA) followed by NGS metagenomic analysis in the detection of human immunodeficiency virus (HIV) and herpes simplex virus (HSV) in cerebrospinal fluid (CSF). Moreover, we analyzed the contamination background. We detected 10(2) HIV copies and 10(3) HSV copies. The analysis of control samples (two water samples and one CSF sample from an uninfected patient) revealed the presence of human DNA in the CSF sample (91 % of all reads), while the dominating sequences in water were qualified as 'other', related to plants, plant viruses, and synthetic constructs, and constituted 31 % and 60 % of all reads. Bacterial sequences represented 5.9 % and 21.4 % of all reads in water samples and 2.3 % in the control CSF sample. The bacterial sequences corresponded mainly to Psychrobacter, Acinetobacter, and Corynebacterium genera. In conclusion, Ribo-SPIA amplification followed by NGS metagenomic analysis is sensitive for detection of RNA and DNA viruses. Contamination seems common and thus the results should be confirmed by other independent methods such as RT-PCR and PCR. Despite these reservations, NGS seems to be a promising method for the diagnosis of viral infections.
A unique strategy, in which invading microorganisms are being caught in web-like structures composed mainly of DNA, involves a recently described phenomenon called NETosis. This process seems to be related to the production of reactive... more
A unique strategy, in which invading microorganisms are being caught in web-like structures composed mainly of DNA, involves a recently described phenomenon called NETosis. This process seems to be related to the production of reactive oxygen species (ROS). In our study, the influence of diphenyleneiodonium chloride (DPI), which diminishes ROS production, was assessed in the context of neutrophil extracellular trap (NET) release. According to protocol, two distinguished procedures were compared, the first one involving DPI elimination from sample before cell activation and the second one proceeding without the step of inhibitor washout. The kinetics of DNA release was monitored by fluorometric assay, and NET formation was observed by fluorescent microscopy. The addition of DPI to the sample led to a reduction of extracellular DNA release. The strongest inhibition was noticed after treatment with 10 μM DPI, which was removed from medium before stimulation with phorbol-12-myristate-13...
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Research Interests: Adolescent, Tuberculosis, Humans, Child, Mycobacterium tuberculosis, and 16 moreFemale, Male, Infant, Differential Diagnosis, Medical Physiology, Aged, Middle Aged, Adult, Biological markers, Sensitivity and Specificity, Pulmonary Tuberculosis, Reference Values, Enzyme Linked Immunosorbent Assay, immunoglobulin G, Immunoglobulin A, and Child preschool
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Identification of submicroscopic chromosomal aberrations, as a cause of structural malformations, is currently performed by MLPA (multiplex ligation-dependent probe amplification) or array CGH (array comparative genomic hybridization)... more
Identification of submicroscopic chromosomal aberrations, as a cause of structural malformations, is currently performed by MLPA (multiplex ligation-dependent probe amplification) or array CGH (array comparative genomic hybridization) techniques. The aim of this study was the evaluation of diagnostic usefulness of MLPA and array CGH in patients with congenital malformations or abnormalities (at least one major or minor birth defect, including dysmorphism) with or without intellectual disability or developmental delay and the optimization of genetic counseling in the context of the results obtained. The MLPA and array CGH were performed in 91 patients diagnosed with developmental disorders and major or minor congenital anomalies. A total of 49 MLPA tests toward common microdeletion syndromes, 42 MLPA tests for subtelomeric regions of chromosomes, two tests for common aberrations in autism, and five array CGH tests were performed. Eight (9 %) patients were diagnosed with microdeletion MLPA, four (4 %) patients with subtelomeric MLPA, one (1 %) patient with autism MLPA. Further three (3 %) individuals had rearrangements diagnosed by array CGH. Altogether, chromosomal microaberrations were found in 16 patients (17 %). All the MLPA-detected rearrangements were found to be pathogenic, but none detected with array CGH could unequivocally be interpreted as pathogenic. In patients with congenital anomalies, the application of MLPA and array CGH techniques is efficient in detecting syndromic and unique microrearrangements. Consistent pre-MLPA test phenotyping leads to better post-test genetic counseling. Incomplete penetrance and unknown inheritance of detected variants are major issues in clinical interpretation of array CGH data.
Photodynamic therapy (PDT) of cancer is an efficient and promising therapeutic modality approved for the treatment of several types of tumors and non-malignant diseases. It involves administration of a non-toxic photosensitizer followed... more
Photodynamic therapy (PDT) of cancer is an efficient and promising therapeutic modality approved for the treatment of several types of tumors and non-malignant diseases. It involves administration of a non-toxic photosensitizer followed by illumination of the tumor site with a harmless visible light. A light activated photosensitizer can transfer its energy directly to molecular oxygen, leading to production of highly toxic reactive oxygen species (ROS). Antitumor effects of PDT result from the combination of three independent mechanisms involving direct cytotoxicity to tumor cells, destruction of tumor vasculature and induction of the acute local inflammatory response. PDT-mediated inflammatory reaction is accompanied by tumor infiltration of the leukocytes, enhanced production of pro-inflammatory factors and cytokines. Photodynamic therapy is able to effectively stimulate both the innate and the adaptive arm of the immune system. In consequence, this regimen can lead to developmen...
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Inflammation of forebrain and hindbrain nuclei controlling the sympathetic nervous system (SNS) outflow from the brain to the periphery represents an emerging concept of the pathogenesis of neurogenic hypertension. Angiotensin II (Ang-II)... more
Inflammation of forebrain and hindbrain nuclei controlling the sympathetic nervous system (SNS) outflow from the brain to the periphery represents an emerging concept of the pathogenesis of neurogenic hypertension. Angiotensin II (Ang-II) and prorenin were shown to increase production of reactive oxygen species and pro-inflammatory cytokines (interleukin-1 beta (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α)) while simultaneously decreasing production of interleukin-10 (IL-10) in the paraventricular nucleus of the hypothalamus and the rostral ventral lateral medulla. Peripheral chronic inflammation and Ang-II activity seem to share a common central mechanism contributing to an increase in sympathetic neurogenic vasomotor tone and entailing neurogenic hypertension. Both hypertension and obesity facilitate the penetration of peripheral immune cells in the brain parenchyma. We suggest that renin-angiotensin-driven hypertension encompasses feedback and feedforward mec...
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The aim of the study was to compare humoral immune response against various mycobacterial antigens in TB and MOTT vs healthy control group. 350 serum samples from TB patients, 20 samples from MOTT patients and 58 samples from healthy... more
The aim of the study was to compare humoral immune response against various mycobacterial antigens in TB and MOTT vs healthy control group. 350 serum samples from TB patients, 20 samples from MOTT patients and 58 samples from healthy donors were examined. ELISA detecting IgG, IgA and IgM against antigens: 38 kDa and 16 kDa, 38kDa and lipoarabinomannan, and A-60 were used. Mean IgG level was higher in TB compared to healthy controls (p<0,001). Mean IgG level against 38kDa and 38 + 16 kDa mycobacterial antigens was higher in TB than in MOTT group. Mean level of the IgG, IgA and IgM antibodies against LAM was higher in MOTT compared to TB patients. In all subgroups person-to-person heterogeneity of antigen recognition was observed. Humoral immune response to recombinant mycobacterial antigens significantly differs in TB and MOTT patients.
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The immune system response and inflammation play a key role in brain injury during and after a stroke. The acute immune response is responsible for secondary brain tissue damage immediately after the stroke, followed by immunosuppression... more
The immune system response and inflammation play a key role in brain injury during and after a stroke. The acute immune response is responsible for secondary brain tissue damage immediately after the stroke, followed by immunosuppression due to sympathetic nervous system activation. The latter increases risk of infection complications, such as pneumonia. The pneumonia-related inflammatory state can release a bystander autoimmune response against central nervous system antigens, thereby initiating a vicious circle. The aim of this review is to summarize the relationship between ischemic stroke, sympathetic nervous system activation and pulmonary infection.
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Wegener's granulomatosis (WG) is characterized histologically by necrotizing granulomatous angitis that most commonly involves the upper, lower respiratory tract and kidneys, but may affect any organ system. Otolaryngological... more
Wegener's granulomatosis (WG) is characterized histologically by necrotizing granulomatous angitis that most commonly involves the upper, lower respiratory tract and kidneys, but may affect any organ system. Otolaryngological manifestations are frequent and diverse but subglottic stenosis and tracheal stenosis are less common. The aim of the study was to assess the clinical features and the response to treatment in WG patients with subglottic or tracheal stenosis. The disease activity at the time of examination was scored in 55 patients with WG (29 females, 26 males) according to clinical, serological, radiological and bronchoscopic findings: subglottic and tracheal stenosis were observed in 9% and 5% of WG patients, respectively. CT scans of the larynx and trachea showed mucosal thickening extended 3-4 cm below the vocal cords in three and the thyroid cartilage in one patient. The degree of narrowing of the axial luminal diameter ranged 50-90%. Mechanical dilation of the stenos...
Research Interests: Humans, Female, Male, Middle Aged, Adult, and 3 moreTracheal stenosis, X ray Computed Tomography, and Glottis
Shiga toxin-producing E. coli O157 and non-O157 are important emergance pathogens that can cause diarrhea and hemorrhagic colitis with life-threatening complications, such as hemolytic uremic syndrome (HUS). A few cases of EHEC infections... more
Shiga toxin-producing E. coli O157 and non-O157 are important emergance pathogens that can cause diarrhea and hemorrhagic colitis with life-threatening complications, such as hemolytic uremic syndrome (HUS). A few cases of EHEC infections are documented per year in Poland. Among them only one patient with EHEC O157 infection developed HUS. We characterized the first VTEC non-O157 strain isolated from child with HUS in Poland. The VTEC O111 strain produced Stx2 which was cytotoxic for Vero cell. Using DNA microarray analysis we have found set of virulence genes in VTEC O111 strain as: stx2A, stx2B, ehly, eae, tir tccP espA, espJ, cif nleA, nleB, lpfA, iha, efa1, cba. The strain was fenotypic resistant to streptomycin, tetracyclin and sulphonamides (strA, tetA, sul2 genes were detected).
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Obesity is a multifactor disease with a very complicated etiology. Genetic factors play an important role in the development of primary obesity. They may be responsible for up to 40% of causes leading to obesity. There are a great number... more
Obesity is a multifactor disease with a very complicated etiology. Genetic factors play an important role in the development of primary obesity. They may be responsible for up to 40% of causes leading to obesity. There are a great number of genes affecting food intake and energy expenditure. Serious consequences accompanying obesity, e.g., type 2 diabetes and lipid abnormalities may be caused by increased level of proinflammatory cytokines, such as IL-1, IL-6, and TNF. It is possible that polymorphisms located in cytokine genes affect the level of protein expression. It is known that IL-6 plays a role in lipid metabolism and energy expenditure. The polymorphism found in point 174 (G174C) of a promoter region of IL-6 gene affects the level of interleukin-6 expression and, consequently, may lead to obesity and correlated conditions.
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The resistance of T lymphocytes to Fas-mediated apoptosis is an important feature of atopic asthma. The only effective causative treatment of atopic diseases is immunotherapy. Clinical efficacy of sublingual immunotherapy (SLIT) has been... more
The resistance of T lymphocytes to Fas-mediated apoptosis is an important feature of atopic asthma. The only effective causative treatment of atopic diseases is immunotherapy. Clinical efficacy of sublingual immunotherapy (SLIT) has been already proven, but there is still limited number of studies on its influence on lymphocytes function. The aim of the study was to evaluate whether SLIT could restore the sensitivity of asthmatic T cells to undergo Fas-mediated apoptosis. Peripheral blood was collected from 12 patients aged 8 ±2 years suffering from atopic asthma and undergoing sublingual specific immunotherapy. To evaluate sensitivity to Fas-mediated apoptosis, the blood was transmitted to sterile tubes and mixed with purified monoclonal antibody anti-CD95. After incubation, leukocytes were stained with Annexin V, propidium iodide, and monoclonal antibody against CD2 conjugated with phycoerythrin-cyanin 5.1, and then analyzed with flow cytometry. The procedure was repeated for each...
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Th1 response is known to play a dominant role in the resistance to tuberculosis. Nevertheless, IFN gamma levels are frequently increased in tuberculous patients, especially at the site of the disease. It is also possible that the shift... more
Th1 response is known to play a dominant role in the resistance to tuberculosis. Nevertheless, IFN gamma levels are frequently increased in tuberculous patients, especially at the site of the disease. It is also possible that the shift toward Th2 response is responsible for the loss of resistance. The aim of this study was to compare the Th1 function of peripheral blood cells and the levels of antimycobacterial antibodies in the serum of culture positive tuberculosis patients and healthy tuberculosis (Tb) contacts. The correlation between the levels of antimycobacterial antibodies and Th1 function of blood cells was also evaluated. The material consisted of 51 tuberculous patients and 20 healthy persons, close contacts of tuberculosis patients. The ability of peripheral blood cells to secrete IFN gamma and IL-2 was estimated in whole blood cultures with PHA, PWM and tuberculin. The levels of IFN gamma and IL-2 in the supernatants of cultures was estimated via a commercial ELISA test...
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The effects of low-level laser light irradiation are debatable and the mechanisms of its action are still unclear. This study was conducted to test the effects of low-level laser irradiation on human blood cells: erythrocytes,... more
The effects of low-level laser light irradiation are debatable and the mechanisms of its action are still unclear. This study was conducted to test the effects of low-level laser irradiation on human blood cells: erythrocytes, granulocytes, and lymphocytes. Whole blood obtained by phlebotomy was irradiated at 632.8 nm by using energy fluences 0.6 J/cm2. An analysis of blood gases revealed an increase in PO2 and SaO2 (P<0.001) in irradiated blood. No shifts in PCO2 and pH were recorded. Spontaneous synthesis of DNA in T and B blood lymphocytes decreased significantly after laser irradiation (P<0.02 and P<0.04, respectively). Phytohemagglutinin (PHA)-induced proliferation of T cells and SAC proliferation of B cells, expressed as a stimulation index, were statistically higher in the samples of irradiated than in non-irradiated blood (P<0.01). Chemiluminescence of fMLP-stimulated granulocytes from irradiated blood increased in comparison with non-irradiated samples (P<0.0...
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The goal of the study was to evaluate IgG, IgA and IgM mediated humoral immune response against 38kDa and 16 kDa or 38kDa and LAM mycobacterial antigens in pleural, pericardial or cerebrospinal fluid from patients with tuberculosis (TB)... more
The goal of the study was to evaluate IgG, IgA and IgM mediated humoral immune response against 38kDa and 16 kDa or 38kDa and LAM mycobacterial antigens in pleural, pericardial or cerebrospinal fluid from patients with tuberculosis (TB) and to compare to non-tuberculous controls (NTB). 30 cerebrospinal fluids (CSF) (16 TB pts and 14 NTB pts), 17 pericardial fluids (6 TB and 11 NTB) and 20 pleural fluids (7 TB and 13 NTB) were examined. Commercially available ELISA-based assays (Pathozyme Tb complex plus, Myco G, A and M--Omega Diagnostic) were used. Tests were performed and cut off established according to manufacturer instruction. Mean IgG level against 38 + 16kDa was significantly higher in neurotuberculosis group compared to control (p<0.05). Sensitivity of the test in detecting neurotuberculosis was of 42% and specificity of 96%. Mean IgG, IgA and IgM against 38kDa + LAM level was higher in TB group compared to NTB in CSF. No difference was observed between TB and NTB group i...
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Research Interests: Bariatric Surgery, Atherosclerosis, Humans, Weight Loss, Female, and 13 moreInborn errors of metabolism, Follow-up studies, Body Mass Index, Obesity Surgery, Clinical Sciences, Middle Aged, Adiponectin, Adult, Public health systems and services research, Matrix Metalloproteinases, Biological markers, Plasminogen Activator Inhibitor, and Case Control Studies
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Disruption of monoamines metabolism leads to diverse manifestations, including developmental, movement and respiratory dysfunctions. We aimed to correlate clinical phenotypes of 55 children with neurodevelopmental disorders with dopamine... more
Disruption of monoamines metabolism leads to diverse manifestations, including developmental, movement and respiratory dysfunctions. We aimed to correlate clinical phenotypes of 55 children with neurodevelopmental disorders with dopamine (HVA) and serotonin (5-HIIA) metabolites in CSF. Decreased level of at least one metabolite was documented in 49.1% patients. Both metabolites were significantly lower in progressive disorder and extrapyramidal syndrome (p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05). HVA was significantly lower in hypokinetic and regulatory disorders (p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05). In univariate analysis, only progressive course, extrapyramidal syndrome and dystonia were significantly associated with decreased 5-HIAA. In multivariate regression only progressive course remained significant (p=0.005). Progressive disease, extrapyramidal syndrome, dystonia, tremor and rigidity were positively associated with low HVA. In multivariate analysis only: progressive course and rigidity remained significant. Progressive/rigid phenotype carries a high risk of monoamines deficiency, strongly implying need for their analysis. Psychomotor delay with epilepsy and hypotonia is rarely linked to low monoamines level. Irrespective of final diagnosis, different clinical presentations may be associated with impaired monoamines turnover.
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Neurotransmitters (NTs) play a central role in the efficient communication between neurons necessary for normal functioning of the nervous system. NTs can be divided into two groups: small molecule NTs and larger neuropeptide NTs.... more
Neurotransmitters (NTs) play a central role in the efficient communication between neurons necessary for normal functioning of the nervous system. NTs can be divided into two groups: small molecule NTs and larger neuropeptide NTs. Inherited disorders of NTs result from a primary disturbance of NTs metabolism or transport. This group of disorders requires sophisticated diagnostic procedures. In this review we discuss disturbances in the metabolism of tetrahydrobiopterin, biogenic amines, γ-aminobutyric acid, foliate, pyridoxine-dependent enzymes, and also the glycine-dependent encephalopathy. We point to pathologic alterations of proteins involved in synaptic neurotransmission that may cause neurological and psychiatric symptoms. We postulate that synaptic receptors and transporter proteins for neurotransmitters should be investigated in unresolved cases. Patients with inherited neurotransmitters disorders present various clinical presentations such as mental retardation, refractory seizures, pyramidal and extrapyramidal syndromes, impaired locomotor patterns, and progressive encephalopathy. Every patient with suspected inherited neurotransmitter disorder should undergo a structured interview and a careful examination including neurological, biochemical, and imaging.
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The ability to occur in a persistent form and to inhibit apoptosis in host cells are the features of Chlamydia pneumoniae which may be associated with adenoid hypertrophy. This study aimed to (1) demonstrate the presence of C. pneumoniae... more
The ability to occur in a persistent form and to inhibit apoptosis in host cells are the features of Chlamydia pneumoniae which may be associated with adenoid hypertrophy. This study aimed to (1) demonstrate the presence of C. pneumoniae DNA in the adenoid in children qualified for adenoidectomy, (2) evaluate the correlation between the presence of C. pneumoniae in the adenoid and the child&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s age, the size of adenoid, the incidence of recurrent respiratory tract infections, (3) demonstrate the co-occurrence of C. pneumoniae and typical bacteria in the adenoid tissue. A group of 200 children aged from 2 to 16 years (mean age 6.4) undergoing adenoidectomy from February 2010 to May 2011 were enrolled to the study. Adenoid tissue removed during planned adenoidectomy was analyzed for the presence of C. pneumoniae by real-time PCR and for the presence of typical bacteria by bacteriological culture. Statistical analysis was conducted using SPSS PASW Statistics 18 software. C. pneumoniae was detected in the adenoid in 5.5% of the 200 children. Positive results were obtained most frequently (24.1%, 7/29) in the eldest group (10-16 years). In PCR positive children the degree of choanal narrowing by an adenoid tended to be larger than in PCR negative children. The occurrence of C. pneumoniae in the adenoid was not associated with a presence of a specific typical bacterium. The presence of C. pneumoniae in an adenoid was most frequently detected in the children of 10-16 years qualified for adenoidectomy.
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Extrinsic allergic alveolitis (EAA) caused by inhaled organic environmental allergens can progress to a fibrotic end-stage lung disease. Neovascularization plays an important role in pathogenesis of pulmonary fibrosis. The aim of this... more
Extrinsic allergic alveolitis (EAA) caused by inhaled organic environmental allergens can progress to a fibrotic end-stage lung disease. Neovascularization plays an important role in pathogenesis of pulmonary fibrosis. The aim of this study was to assess the effect of sera from EAA patients on the angiogenic capability of normal peripheral human mononuclear cells (MNC) in relation to the clinical, radiological, and functional changes. The study population consisted of 30 EAA patients and 16 healthy volunteers. Routine pulmonary function tests were undertaken using ERS standards. As an angiogenic test, leukocyte-induced angiogenesis assay according to Sidky and Auerbach was used. Compared with sera from healthy volunteers, sera from our EAA patients significantly stimulated angiogenesis (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001). However, sera from healthy donors also stimulated angiogenesis compared to PBS (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001). No correlation was found between serum angiogenic activity and clinical symptoms manifested by evaluated patients. A decrease in DLco and in lung compliance in EAA patients was observed but no significant correlation between pulmonary functional tests and serum angiogenic activity measured by the number of microvessels or an angiogenesis index was found. However, the proangiogenic effect of sera from EAA patients differed depending on the stage of the disease and was stronger in patients with fibrotic changes. The present study suggests that angiogenesis plays a role in the pathogenesis of EAA. It could be possible that the increase in the angiogenic activity of sera from EAA patients depends on the phase of the disease.
Research Interests: Adolescent, Humans, Pulmonary fibrosis, Mice, Female, and 9 moreAnimals, Male, Young Adult, Lung, Aged, Middle Aged, Adult, Pulmonary Function, and SERUM
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Specific immunotherapy is a causative, effective asthma treatment. Since subcutaneous immunotherapy (SCIT) has been found as partially questionable due to its side effects, sublingual immunotherapy (SLIT) arouse wide interest. SLIT very... more
Specific immunotherapy is a causative, effective asthma treatment. Since subcutaneous immunotherapy (SCIT) has been found as partially questionable due to its side effects, sublingual immunotherapy (SLIT) arouse wide interest. SLIT very seldom causes acute ...
The eosin-5′-maleimide (EMA) binding test is a flow cytometric test widely used to detect hereditary spherocytosis (HS). EMA binds to plasma membrane proteins of red blood cells (RBCs), mainly to band 3 protein. The mean fluorescence of... more
The eosin-5′-maleimide (EMA) binding test is a flow cytometric test widely used to detect hereditary spherocytosis (HS). EMA binds to plasma membrane proteins of red blood cells (RBCs), mainly to band 3 protein. The mean fluorescence of EMA-stained RBCs in HS patients is lower when compared with control RBCs due to the decreased amount of target proteins. EMA dye in aqueous solution is sensitive to light and high temperature. Its fluorescence can decrease when exposed to light or ambient temperatures higher than 4°C. The aim of the study was to evaluate the stability of fluorescence readings of EMA-labeled RBCs over a period of 24 h. The EMA test was performed in peripheral blood from 35 patients with microcytic anemia (five with HS, and 30 without HS). Peripheral blood samples were stained immediately after blood collection and analyzed using a flow cytometer at three time points: 0, after 1 and 24 h of storage at 4°C in the darkness. The results are presented as the percentage of normal control RBCs fluorescence. Flow cytometric studies were performed with Cytomics FC500 (Beckman Coulter, USA). In HS patients the mean result of the test reached 66.72%±9.26% of normal controls, and in non-HS patients the EMA result was 99.48%±5.03% of normal control cells. The results of patients with HS were 66.72%±9.26%, 66.90%±10.24% and 67.86%±11.31% at 0 h, and after 1 and 24 h of storage, respectively. The results obtained from non-HS patients at time 0, after 1 and 24 h of storage reached 99.48%±5.03%, 99.49%±5.34% and 99.78%±6.13%, respectively. There was no difference between the results from each time point in samples from patients with or without HS. Results of the EMA binding test do not depend on storage time of stained samples when stored at 4°C up to 24 h after staining.
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Eighteen patients (12 female) with Wegener&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s granulomatosis (WG) treated from 1982-2001 were studied. Mean duration of patients&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; symptoms... more
Eighteen patients (12 female) with Wegener&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s granulomatosis (WG) treated from 1982-2001 were studied. Mean duration of patients&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; symptoms prior to diagnosis was 13.5 months, and mean age 41.2. Lung biopsy confirmed WG in 5 patients, nasal biopsy was made in 12, endobronchial in 3. In 11 of 14 patients cANCAs were found. Prednisone and 2 mg/kg/day doses of oral cyclophosphamide were used to treat 17 patients, prednisone was used alone for 1. Patients with severe symptoms were given i.v. &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;pulse&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; methylprednisolone. Mean duration of treatment was 34.8 months and of follow-up, 71.9 months. A total of 4 patients died: 3 from complete renal failure. Four patients still undergo treatment, improving markedly. Complete remission was achieved in 6 patients. A total of 3 patients relapsed and are recently treated. In another one chronic renal failure developed. Three of them had moderate symptoms at diagnosis but were treated inadequately. manifestation of WG and severity of symptoms may be different among patients. ANCA is not universally present in all patients with WG. The absence of ANCA does not exclude WG diagnosis. Complete remission can be achieved in WG patients, including the severely ill, if adequate therapy is given. Moderate symptoms at presentation do not justify less intensive treatment.