e18576 Background: We prospectively studied 90 patients of multiple myeloma for the presence of t... more e18576 Background: We prospectively studied 90 patients of multiple myeloma for the presence of translocations involving IgH (14q32) locus, del 13q14 and presence of numerical chromosomal abnormalities. An attempt was made to correlate their effect on outcome and to formulate a risk scoring system. METHODS Purified bone marrow plasma cells, using magnetic activated cell sorter (MACS) with CD 138 micro beads were used. Fluorescent in situ Hybridization was used to detect IgH (14q32) translocations and del 13q14, while numerical chromosomal abnormalities were detected by conventional cytogenetics. RESULTS Patients' median age was 55 years (range, 34 to 75 years) and 67 were males (M: F: 2.9: 1). 24 patients had International staging system (ISS) stage I, stage II- 46 and 20 patients had stage III. On conventional cytogenetics metaphases were obtained in 53 (58%) patients; with numerical chromosomal abnormalities in 21 of 53 (40%), these included- hyperdiploid in 16 and hypodiploid-5. Remaining patients had normal metaphases. del 13q14 and t(14q32) were present in 51 (56%) and 75 (83%) patients, respectively. 65/90 (72%) patients received therapy with novel molecules and were evaluated using EBMT response criteria. Absence of del 13q14 was associated with higher response rate (23/26 vs 26/39, P <0.04) and better overall survival (79% vs 67%, p =0.07) and event free survival (70% vs 37%, p <0.03) at two years. Based on presence (1) or absence (0) of del 13q14 and ISS (I=0 vs II=0.5 vs III=1), a risk scoring model was obtained. 69% of the patients belonged to low risk category (risk score ≤1) and 31% were in high risk (risk score >1). Patients belonging to low risk had a higher response (91% vs 40%, p<.0001), better overall (77% vs 61%, p<.01) and event free (58% vs 32%, p<.01) survival at two years compared to patients with high risk. CONCLUSIONS del 13q14 and International staging system (ISS) are two important parameters and could be used to prognosticate patients at diagnosis.
Background:Inflammatory responses within the peritoneal cavity may result in endometrial dysfunct... more Background:Inflammatory responses within the peritoneal cavity may result in endometrial dysfunction in women with endometriosis. The true causes of this disease remain poorly understood. It is hypothesized that downstream toll-like receptors (TLRs) inflammatory cytokines in response to pathogens may be associated with endometriosis. So, this study was aimed at evaluating the expression of TLRs signaling and endometriosis-associated inflammatory responses.Methods:Totally, 20 infertile endometriosis patients and 20 normal women undergoing controlled ovarian stimulation were enrolled. The cellular pellet and supernatant were obtained by centrifugation of follicular fluid (FF). Evaluation of TLRs and their signaling pathway gene expression was performed on cellular pellets using quantitative-PCR. The supernatant was used for determination of cytokine protein expression by ELISA. The results are expressed as mean±SEM and a p<0.05 was considered statistically significant.Results:Quantitative-PCR analysis suggested that TLR1, 5, 6, 7, 8, 10, MYD88, NF-ĸB, IL-10 and TGF-β genes expression significantly increased in patients compared to the control group (p<0.05). TLR3, 9, INF-β genes expression was significantly lower in endometriosis than control group (p<0.05). There was no significant difference in the expression of TLR2, TLR4, TIRAP, TRIF, TRAM, and IRF3 between two groups. Also, significant increase in the levels of IL-6, IL-8 and MIF protein in FF of endometriosis group was detected in comparison with normal women (p<0.05).Conclusion:The expression of TLR downstream signaling in the follicular cells can initiate inflammatory responses and changes in the FF cytokine profile which in turn may induce endometriosis and infertility disorder.
International Journal of Clinical Biochemistry and Research, Sep 15, 2022
: Tumor necrosis factor-alpha (TNF-α) is a pro-inflammatory cytokine implicated in various physio... more : Tumor necrosis factor-alpha (TNF-α) is a pro-inflammatory cytokine implicated in various physiological and pathological events. Carnitine is a quaternary amine which plays a significant role in fatty acid oxidation and is reported to produce antiapoptotic effects. Aim of this work was to study the effect of L-Carnitine (LC) on TNF-α induced apoptosis in mice oocytes. : Oocytes were isolated from super ovulated Swiss Albino mice and treated with different concentrations of TNF-α (0.1ng/ml, 1ng/ml, 10ng/ml, 100ng/ml) and LC (0.1mg/ml, 0.3mg/ml, 0.5mg/ml, 1.0 mg/ml). TUNEL Assay was done for the biochemical assessment of apoptosis.: Apoptotic indices with different doses of TNF-α (0.1ng/ml, 1ng/ml, 10ng/ml, 100 ng/ml) were 28.5%, 71.4%, 100%, 42.8% respectively. The concentration of TNF-α that produced the highest apoptotic index was 10ng/ml. LC alone in different doses (0.1mg/ml, 0.3mg/ml, 0.5mg/ml, 1.0 mg/ml) did not elicit any apoptotic signal. Further LC was added in different doses with 10ng/ml TNF-α to study the rate of apoptosis in mice oocytes. Apoptotic index with 10 ng/ml TNF-α and different doses of LC (0.1mg/ml, 0.3mg/ml, 0.5mg/ml, 1 mg/ml) were 25%, 37.5%, 50%, 62.5% respectively. The concentration of LC that reduced the apoptotic index to the maximum was 0.1mg/ml.: Present study could demonstrate the anti-apoptotic effect of LC against apoptotic effects of TNF-α in mice oocytes. The study presents preliminary data suggesting a possible therapeutic role of LC in inflammatory etiologies such as ovarian failure.
We report the seventh family of Fowler like syndrome (proliferative vasculopathy and hydrocephaly... more We report the seventh family of Fowler like syndrome (proliferative vasculopathy and hydrocephaly-hydrencephaly syndrome) and first case from Indian subcontinent. A 35 weeks extremely growth retarded male baby showed enlarged ventricles, thinned out cerebral cortex, diffuse intra-cerebral as well as peri-ventricular calcification, cerebral and corneal vasculopathy, unilateral micro-ophthalmia along with corneal opacity and depressed pulsatile anterior fontanel. This case was different from others concerning association with extreme oligohydramnios (in contrast to polyhydramnios), extreme growth restriction (in contrast to normal growth) and absence of gross muscle hypoplasia. No causative factors like TORCH infection, chromosomal abnormality or positive family history was noted in this case.
Meiotic segregation of chromosomes X,Y and 1 was analyzed by triple colour rapid fluorescent in s... more Meiotic segregation of chromosomes X,Y and 1 was analyzed by triple colour rapid fluorescent in situ hybridization (FISH) with directly labelled probes on 4506 non-decondensed and non-cleaned interphase spermatozoa from four healthy male donors to test the possibility of rapid sperm FISH by omitting the conventional sperm decondensation and cleaning steps. Only 0.15 per cent of sperms were without any signals which suggested high hybridization success. An abnormal number of signals was seen in 1.6 per cent of sperms. Chromosome specific as well as donor specific segregation error was seen similar to previous reports. There was a wide variation in the ratio of normal X and Y bearing sperm from donor to donor. This study indicated that for segregation studies sperm FISH can be carried out in three hours with directly labelled probes without the steps for separation of sperm from somatic cells (cleaning), sperm swelling and sperm DNA decondensation.
Prader Willi syndrome (PWS) most commonly is due to paternal micro-deletion of 15q11-q13. Althoug... more Prader Willi syndrome (PWS) most commonly is due to paternal micro-deletion of 15q11-q13. Although PWS is not a rare condition, mosaic micro-deletion cases are reported rarely. FISH using PWS micro-deletion probe is the most useful method to detect deletion including mosaicism. In this report we describe a female child with clinical features of atypical PWS and FISH analysis showing mosaicism for deletion in the PWS critical region. This is first mosaic deletion case of PWS from Indian subcontinent.
We have evaluated the suitability of different formalin fixed paraffin embedded tissues i.e., bra... more We have evaluated the suitability of different formalin fixed paraffin embedded tissues i.e., brain, bone, liver and placenta, for fluorescence in situ hybridization (FISH) efficiency and chromosomal ploidy detection employing directly labelled repetitive sequence probes for chromosome X, Y and 1. The study was carried out on four foetal autopsy specimens. Cells from 50 mu thick tissue sections were dissociated before performing mono and multicolour FISH with directly labelled probes. Hybridization efficiency was maximum with brain tissue (81, 55 and 24% for mono, dual and triple colour FISH, respectively), followed by bone (45 and 34% for mono and dual colour FISH), liver (38, 19 and 0% for mono, dual and triple colour FISH) and placenta (14, 5 and 0% for mono, dual and triple colour FISH). These results indicated that brain is the most efficient material followed by bone, liver placenta for chromosome ploidy detection by FISH in formalin fixed tissues.
Global journal of human genetics & gene therapy, Feb 1, 2014
ABSTRACT Disorder of sex development is defined as developmental abnormality in which determinati... more ABSTRACT Disorder of sex development is defined as developmental abnormality in which determination and/or differentiation of chromosomal, gonadal or phenotypic sex is abnormal. This is a common disorder and a precise diagnosis is very important for right management to prevent future psychosexual problems. The aim of this study was to evaluate the clinical features documented, investigative procedures followed and corrective interventions adopted during the course of the study and recommend how these cases could be managed satisfactorily without high end investigative support. This study describes the spectrum of cases of disorder of sex development in a referral tertiary care hospital in North India. The medical records of 63 patients presenting with disorder of sex development over a period of three years were reviewed. On the basis of meticulous clinical evaluation and minimal investigations (chromosomal, hormonal, biochemical, radiological, genitoscopy and laparoscopy/laparotomy/ gonadal biopsy) cases were categorized as 46,XY disorder of sex development (40 cases/63.5%), 46,XX disorder of sex development (14 cases/22.2%) and gonadal differentiation disorder of sex development (ovotesticular disorder of sex development &amp; gonadal dysgenesis; 9 cases/14.3%). Ambiguous genitalia/precocious puberty was the main presentation in 46,XX disorder of sex development whereas ambiguous genitalia/under development of breast or breast development at puberty was the main presentation in 46,XY disorder of sex development. Ambiguous genitalia was the major presenting complaint of cases of gonadal differentiation disorder of sex development. There were three peaks of age at presentation; infancy, childhood and postpubertal period. Mullerian ducts remnants were found in nine (22.5%) of 46,XY disorder of sex development. Male like external genitalia with bilateral cryptorchidism were also evident with 46,XX disorder of sex development. Sex assignment/reassignment was based mainly on virilization of external genitalia (phallus size) and testosterone response (excepting 46,XX disorder of sex development). The study concludes that with meticulous clinical evaluation and base line investigations cases of disorder of sex development can be managed well
Indian Journal of Pathology and Microbiology, 2020
Increasing HER-2/neu resistance in gastric carcinoma has encouraged search for new biomarkers for... more Increasing HER-2/neu resistance in gastric carcinoma has encouraged search for new biomarkers for targeted therapy. Cellular mesenchymal epithelial transition (C-MET) is one such tyrosine kinase inhibitor proposed for personalized salvage treatment. We determined frequency of C-MET gene copy number variation (CNV) by Fluorescent in-situ hybridization (FISH) in gastric adenocarcinoma (GAC) and sought its correlation with conventional clinicopathologic parameters. Dual-coloured FISH was done on 32 GAC cases. C-MET gene and centromere 7 signals were counted under fluorescent microscope and ratio was calculated for each case. Correlation between C-MET CNV and conventional clinic-pathologic parameters was done by Fischer exact test. CNV was identified in the form of amplification and polysomy (3.1% each) and associated with poorer prognostic parameters. Our pilot study highlights limited subset of patients that may benefit from anti-C-MET-targeted therapy and thus could be a novel biomarker for targeted intervention in GAC.
Primary testicular failure and its subtypes Etiology Biomarkers Principles of diagnostic evaluati... more Primary testicular failure and its subtypes Etiology Biomarkers Principles of diagnostic evaluation and management
Sertoli cell only syndrome (SCOS) is characterized by complete absence of germ cells in seminifer... more Sertoli cell only syndrome (SCOS) is characterized by complete absence of germ cells in seminiferous tubules of testis. SCOS is multifactorial but genetic factor play a major role in pathogenesis of the disorder with idiopathic origin. Genetic factor majorly includes sex chromosomal aneuploidy and Yq Microdeletion. But a large number of cases are still idiopathic. The study aimed to evaluate the genomic imbalances (CNVs and LOH) in idiopathic SCOS patients. The study is based on 28 apparent idiopathic SCOS cases and 10 controls. Molecular cytogenetic techniques viz., FISH, STS-Multiplex PCR and Affymetrix cytoscan microarray (750K) were used. The microarray screened whole genomic imbalances in DNA from peripheral blood for 25 cases (excluded Klinefelter syndrome patients) and testicular FNAC sample for 2 cases. High FSH and low Inhibin B were observed in cases than controls groups. Four cases of sex chromosomal abnormality (i.e., three non-mosaic 47,XXY males and one non-mosaic 46,XX male) as well as four cases of Yq microdeletion (i.e., three cases with AZFc deletion and one case with complete AZFa, b and c deletion) were identified. Microarray detected unbalanced translocation of two segments of Y-chromosome i.e., Yp11.31-p11.2 (∼4.o mb region, involving SRY) and Yp11.2 (∼2.5 mb region) on X-chromosome in XX male. Also, loss of segment on same X-chromosome involving PAR1 region was identified. We have identified both autosomal and sex chromosomal CNVs (recurrent as well as private) involving candidate genes like SYCE1, ZFPM2, SRPK1, DAZ1, BPY2, HSFY1, VCY1 etc. All these CNVs possibly associated with SCOS pathogenesis. CNVs identified in cases that were already reported as pathogenic variant in clinical database DECIPHER. Microarray also detected many LOH (all autosomal, >3.0 mb size) that covered genes with spermatogenesis related function. The mechanism of action of LOH in pathogenesis of SCOS is still unravelled. CNVs and LOH related to spermatogenesis identified from two different sample types (blood vs. testicular tissue) were discordant. This study should be extended for larger cohort of patients.
e18576 Background: We prospectively studied 90 patients of multiple myeloma for the presence of t... more e18576 Background: We prospectively studied 90 patients of multiple myeloma for the presence of translocations involving IgH (14q32) locus, del 13q14 and presence of numerical chromosomal abnormalities. An attempt was made to correlate their effect on outcome and to formulate a risk scoring system. METHODS Purified bone marrow plasma cells, using magnetic activated cell sorter (MACS) with CD 138 micro beads were used. Fluorescent in situ Hybridization was used to detect IgH (14q32) translocations and del 13q14, while numerical chromosomal abnormalities were detected by conventional cytogenetics. RESULTS Patients' median age was 55 years (range, 34 to 75 years) and 67 were males (M: F: 2.9: 1). 24 patients had International staging system (ISS) stage I, stage II- 46 and 20 patients had stage III. On conventional cytogenetics metaphases were obtained in 53 (58%) patients; with numerical chromosomal abnormalities in 21 of 53 (40%), these included- hyperdiploid in 16 and hypodiploid-5. Remaining patients had normal metaphases. del 13q14 and t(14q32) were present in 51 (56%) and 75 (83%) patients, respectively. 65/90 (72%) patients received therapy with novel molecules and were evaluated using EBMT response criteria. Absence of del 13q14 was associated with higher response rate (23/26 vs 26/39, P <0.04) and better overall survival (79% vs 67%, p =0.07) and event free survival (70% vs 37%, p <0.03) at two years. Based on presence (1) or absence (0) of del 13q14 and ISS (I=0 vs II=0.5 vs III=1), a risk scoring model was obtained. 69% of the patients belonged to low risk category (risk score ≤1) and 31% were in high risk (risk score >1). Patients belonging to low risk had a higher response (91% vs 40%, p<.0001), better overall (77% vs 61%, p<.01) and event free (58% vs 32%, p<.01) survival at two years compared to patients with high risk. CONCLUSIONS del 13q14 and International staging system (ISS) are two important parameters and could be used to prognosticate patients at diagnosis.
Background:Inflammatory responses within the peritoneal cavity may result in endometrial dysfunct... more Background:Inflammatory responses within the peritoneal cavity may result in endometrial dysfunction in women with endometriosis. The true causes of this disease remain poorly understood. It is hypothesized that downstream toll-like receptors (TLRs) inflammatory cytokines in response to pathogens may be associated with endometriosis. So, this study was aimed at evaluating the expression of TLRs signaling and endometriosis-associated inflammatory responses.Methods:Totally, 20 infertile endometriosis patients and 20 normal women undergoing controlled ovarian stimulation were enrolled. The cellular pellet and supernatant were obtained by centrifugation of follicular fluid (FF). Evaluation of TLRs and their signaling pathway gene expression was performed on cellular pellets using quantitative-PCR. The supernatant was used for determination of cytokine protein expression by ELISA. The results are expressed as mean±SEM and a p<0.05 was considered statistically significant.Results:Quantitative-PCR analysis suggested that TLR1, 5, 6, 7, 8, 10, MYD88, NF-ĸB, IL-10 and TGF-β genes expression significantly increased in patients compared to the control group (p<0.05). TLR3, 9, INF-β genes expression was significantly lower in endometriosis than control group (p<0.05). There was no significant difference in the expression of TLR2, TLR4, TIRAP, TRIF, TRAM, and IRF3 between two groups. Also, significant increase in the levels of IL-6, IL-8 and MIF protein in FF of endometriosis group was detected in comparison with normal women (p<0.05).Conclusion:The expression of TLR downstream signaling in the follicular cells can initiate inflammatory responses and changes in the FF cytokine profile which in turn may induce endometriosis and infertility disorder.
International Journal of Clinical Biochemistry and Research, Sep 15, 2022
: Tumor necrosis factor-alpha (TNF-α) is a pro-inflammatory cytokine implicated in various physio... more : Tumor necrosis factor-alpha (TNF-α) is a pro-inflammatory cytokine implicated in various physiological and pathological events. Carnitine is a quaternary amine which plays a significant role in fatty acid oxidation and is reported to produce antiapoptotic effects. Aim of this work was to study the effect of L-Carnitine (LC) on TNF-α induced apoptosis in mice oocytes. : Oocytes were isolated from super ovulated Swiss Albino mice and treated with different concentrations of TNF-α (0.1ng/ml, 1ng/ml, 10ng/ml, 100ng/ml) and LC (0.1mg/ml, 0.3mg/ml, 0.5mg/ml, 1.0 mg/ml). TUNEL Assay was done for the biochemical assessment of apoptosis.: Apoptotic indices with different doses of TNF-α (0.1ng/ml, 1ng/ml, 10ng/ml, 100 ng/ml) were 28.5%, 71.4%, 100%, 42.8% respectively. The concentration of TNF-α that produced the highest apoptotic index was 10ng/ml. LC alone in different doses (0.1mg/ml, 0.3mg/ml, 0.5mg/ml, 1.0 mg/ml) did not elicit any apoptotic signal. Further LC was added in different doses with 10ng/ml TNF-α to study the rate of apoptosis in mice oocytes. Apoptotic index with 10 ng/ml TNF-α and different doses of LC (0.1mg/ml, 0.3mg/ml, 0.5mg/ml, 1 mg/ml) were 25%, 37.5%, 50%, 62.5% respectively. The concentration of LC that reduced the apoptotic index to the maximum was 0.1mg/ml.: Present study could demonstrate the anti-apoptotic effect of LC against apoptotic effects of TNF-α in mice oocytes. The study presents preliminary data suggesting a possible therapeutic role of LC in inflammatory etiologies such as ovarian failure.
We report the seventh family of Fowler like syndrome (proliferative vasculopathy and hydrocephaly... more We report the seventh family of Fowler like syndrome (proliferative vasculopathy and hydrocephaly-hydrencephaly syndrome) and first case from Indian subcontinent. A 35 weeks extremely growth retarded male baby showed enlarged ventricles, thinned out cerebral cortex, diffuse intra-cerebral as well as peri-ventricular calcification, cerebral and corneal vasculopathy, unilateral micro-ophthalmia along with corneal opacity and depressed pulsatile anterior fontanel. This case was different from others concerning association with extreme oligohydramnios (in contrast to polyhydramnios), extreme growth restriction (in contrast to normal growth) and absence of gross muscle hypoplasia. No causative factors like TORCH infection, chromosomal abnormality or positive family history was noted in this case.
Meiotic segregation of chromosomes X,Y and 1 was analyzed by triple colour rapid fluorescent in s... more Meiotic segregation of chromosomes X,Y and 1 was analyzed by triple colour rapid fluorescent in situ hybridization (FISH) with directly labelled probes on 4506 non-decondensed and non-cleaned interphase spermatozoa from four healthy male donors to test the possibility of rapid sperm FISH by omitting the conventional sperm decondensation and cleaning steps. Only 0.15 per cent of sperms were without any signals which suggested high hybridization success. An abnormal number of signals was seen in 1.6 per cent of sperms. Chromosome specific as well as donor specific segregation error was seen similar to previous reports. There was a wide variation in the ratio of normal X and Y bearing sperm from donor to donor. This study indicated that for segregation studies sperm FISH can be carried out in three hours with directly labelled probes without the steps for separation of sperm from somatic cells (cleaning), sperm swelling and sperm DNA decondensation.
Prader Willi syndrome (PWS) most commonly is due to paternal micro-deletion of 15q11-q13. Althoug... more Prader Willi syndrome (PWS) most commonly is due to paternal micro-deletion of 15q11-q13. Although PWS is not a rare condition, mosaic micro-deletion cases are reported rarely. FISH using PWS micro-deletion probe is the most useful method to detect deletion including mosaicism. In this report we describe a female child with clinical features of atypical PWS and FISH analysis showing mosaicism for deletion in the PWS critical region. This is first mosaic deletion case of PWS from Indian subcontinent.
We have evaluated the suitability of different formalin fixed paraffin embedded tissues i.e., bra... more We have evaluated the suitability of different formalin fixed paraffin embedded tissues i.e., brain, bone, liver and placenta, for fluorescence in situ hybridization (FISH) efficiency and chromosomal ploidy detection employing directly labelled repetitive sequence probes for chromosome X, Y and 1. The study was carried out on four foetal autopsy specimens. Cells from 50 mu thick tissue sections were dissociated before performing mono and multicolour FISH with directly labelled probes. Hybridization efficiency was maximum with brain tissue (81, 55 and 24% for mono, dual and triple colour FISH, respectively), followed by bone (45 and 34% for mono and dual colour FISH), liver (38, 19 and 0% for mono, dual and triple colour FISH) and placenta (14, 5 and 0% for mono, dual and triple colour FISH). These results indicated that brain is the most efficient material followed by bone, liver placenta for chromosome ploidy detection by FISH in formalin fixed tissues.
Global journal of human genetics & gene therapy, Feb 1, 2014
ABSTRACT Disorder of sex development is defined as developmental abnormality in which determinati... more ABSTRACT Disorder of sex development is defined as developmental abnormality in which determination and/or differentiation of chromosomal, gonadal or phenotypic sex is abnormal. This is a common disorder and a precise diagnosis is very important for right management to prevent future psychosexual problems. The aim of this study was to evaluate the clinical features documented, investigative procedures followed and corrective interventions adopted during the course of the study and recommend how these cases could be managed satisfactorily without high end investigative support. This study describes the spectrum of cases of disorder of sex development in a referral tertiary care hospital in North India. The medical records of 63 patients presenting with disorder of sex development over a period of three years were reviewed. On the basis of meticulous clinical evaluation and minimal investigations (chromosomal, hormonal, biochemical, radiological, genitoscopy and laparoscopy/laparotomy/ gonadal biopsy) cases were categorized as 46,XY disorder of sex development (40 cases/63.5%), 46,XX disorder of sex development (14 cases/22.2%) and gonadal differentiation disorder of sex development (ovotesticular disorder of sex development &amp; gonadal dysgenesis; 9 cases/14.3%). Ambiguous genitalia/precocious puberty was the main presentation in 46,XX disorder of sex development whereas ambiguous genitalia/under development of breast or breast development at puberty was the main presentation in 46,XY disorder of sex development. Ambiguous genitalia was the major presenting complaint of cases of gonadal differentiation disorder of sex development. There were three peaks of age at presentation; infancy, childhood and postpubertal period. Mullerian ducts remnants were found in nine (22.5%) of 46,XY disorder of sex development. Male like external genitalia with bilateral cryptorchidism were also evident with 46,XX disorder of sex development. Sex assignment/reassignment was based mainly on virilization of external genitalia (phallus size) and testosterone response (excepting 46,XX disorder of sex development). The study concludes that with meticulous clinical evaluation and base line investigations cases of disorder of sex development can be managed well
Indian Journal of Pathology and Microbiology, 2020
Increasing HER-2/neu resistance in gastric carcinoma has encouraged search for new biomarkers for... more Increasing HER-2/neu resistance in gastric carcinoma has encouraged search for new biomarkers for targeted therapy. Cellular mesenchymal epithelial transition (C-MET) is one such tyrosine kinase inhibitor proposed for personalized salvage treatment. We determined frequency of C-MET gene copy number variation (CNV) by Fluorescent in-situ hybridization (FISH) in gastric adenocarcinoma (GAC) and sought its correlation with conventional clinicopathologic parameters. Dual-coloured FISH was done on 32 GAC cases. C-MET gene and centromere 7 signals were counted under fluorescent microscope and ratio was calculated for each case. Correlation between C-MET CNV and conventional clinic-pathologic parameters was done by Fischer exact test. CNV was identified in the form of amplification and polysomy (3.1% each) and associated with poorer prognostic parameters. Our pilot study highlights limited subset of patients that may benefit from anti-C-MET-targeted therapy and thus could be a novel biomarker for targeted intervention in GAC.
Primary testicular failure and its subtypes Etiology Biomarkers Principles of diagnostic evaluati... more Primary testicular failure and its subtypes Etiology Biomarkers Principles of diagnostic evaluation and management
Sertoli cell only syndrome (SCOS) is characterized by complete absence of germ cells in seminifer... more Sertoli cell only syndrome (SCOS) is characterized by complete absence of germ cells in seminiferous tubules of testis. SCOS is multifactorial but genetic factor play a major role in pathogenesis of the disorder with idiopathic origin. Genetic factor majorly includes sex chromosomal aneuploidy and Yq Microdeletion. But a large number of cases are still idiopathic. The study aimed to evaluate the genomic imbalances (CNVs and LOH) in idiopathic SCOS patients. The study is based on 28 apparent idiopathic SCOS cases and 10 controls. Molecular cytogenetic techniques viz., FISH, STS-Multiplex PCR and Affymetrix cytoscan microarray (750K) were used. The microarray screened whole genomic imbalances in DNA from peripheral blood for 25 cases (excluded Klinefelter syndrome patients) and testicular FNAC sample for 2 cases. High FSH and low Inhibin B were observed in cases than controls groups. Four cases of sex chromosomal abnormality (i.e., three non-mosaic 47,XXY males and one non-mosaic 46,XX male) as well as four cases of Yq microdeletion (i.e., three cases with AZFc deletion and one case with complete AZFa, b and c deletion) were identified. Microarray detected unbalanced translocation of two segments of Y-chromosome i.e., Yp11.31-p11.2 (∼4.o mb region, involving SRY) and Yp11.2 (∼2.5 mb region) on X-chromosome in XX male. Also, loss of segment on same X-chromosome involving PAR1 region was identified. We have identified both autosomal and sex chromosomal CNVs (recurrent as well as private) involving candidate genes like SYCE1, ZFPM2, SRPK1, DAZ1, BPY2, HSFY1, VCY1 etc. All these CNVs possibly associated with SCOS pathogenesis. CNVs identified in cases that were already reported as pathogenic variant in clinical database DECIPHER. Microarray also detected many LOH (all autosomal, >3.0 mb size) that covered genes with spermatogenesis related function. The mechanism of action of LOH in pathogenesis of SCOS is still unravelled. CNVs and LOH related to spermatogenesis identified from two different sample types (blood vs. testicular tissue) were discordant. This study should be extended for larger cohort of patients.
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