Immunohistochemical evaluation of p53, FHIT, and IGF2 gene expression in esophageal cancer.

The aim of the study was to evaluate the expression of tumor suppressor genes p53, fragile histidine triad gene (FHIT), and an oncogene insulin-like growth factor 2 (IGF2) as prognostic markers in the etiology of esophageal cancer. Immunohistochemistry (IHC) was performed in 39 archival tissue samples of different esophageal pathologies for the three genes. Abnormal p53 expression was maximum in all the cases of squamous cell carcinoma, while IGF2 expression was enhanced in squamous cell carcinoma (81%), adenocarcinoma (100%), and dysplasia of squamous epithelium (75%) samples when compared with normals (50%). To our surprise, 75% of normal tissues did not show FHIT expression, which was also not seen in 40% of dysplasias of squamous epithelium, 33.3% of adenocarcinoma, and 41% of squamous cell carcinoma. To the best of our knowledge, this is the first study evaluating IGF2 by IHC, as well as, correlating it with the expression of the two tumor suppressor genes, p53 and FHIT, in esophageal tissue. p53 expression was threefold higher than normal in dysplasias of squamous epithelium and adenocarcinoma, while it was eightfold higher in squamous cell carcinoma. IGF2 expression was low in normal and dysplasia tissue but was increased 1.97-fold in both types of malignancy. FHIT and p53 expression were well correlated in squamous cell carcinoma, supporting the observation that FHIT regulates and stabilizes p53. Altered/lowered FHIT levels may be a result of exposure to various exogenous agents; however, this could not be assessed in the present study as it was carried out on archival samples. A larger prospective study is warranted to establish the role of exogenous factors in FHIT expression.

Inequities in the deployment of COVID-19 vaccine in the WHO Eastern Mediterranean Region, 2020-2021.

The WHO Eastern Mediterranean Region (EMR) is characterised by a large range in routine immunisation coverage. We reviewed progress in access, deployment efforts, and use of COVID-19 vaccines in the EMR to identify bottlenecks and propose recommendations. We compiled and analysed data reported to WHO regarding the number of vaccines provided emergency use authorisation (EUA) in each country, the number of vaccine doses allocated and delivered by COVAX, the number of vaccine doses received bilaterally, the date of initiation of vaccination, vaccine usage rate and overall vaccination coverage. In June-July and October-November 2021, we conducted two rounds of a regional survey to assess vaccine acceptance and calculated the weighted proportion of individuals who would get vaccinated once a vaccine is available and recommended. We stratified the analysis according to four groups based on their participation status in COVAX, from the highest to lowest income, that is, (1) fully self-financing high-income countries (group 1), (2) fully self-financing upper middle-income countries (group 2), (3) Advance Market Commitment (AMC) countries not eligible to receive Gavi support (group 3) and (4) AMC countries eligible for Gavi support (group 4). As of 31 December 2021, the median number of vaccines provided with EUA was 6 for group 1, 11 for group 2, 8 for group 3 and 9 for group 4. On the same date, COVAX had delivered 179 793 310 doses to EMR countries. Vaccination started on 10 December 2020 in group 1, on 13 December 2020 in group 2, on 30 December 2020 in group 3 and on 20 January 2021 in group 4. The regional acceptance survey (first round) pointed to higher vaccine acceptance in group 1 (96%), than in others, including group 2 (73.9%), group 3 (78.8%) and group 4 (79.3%), with identical patterns in the second round (98%, 78%, 84% and 76%), respectively. Usage of vaccine allocated by COVAX to participating countries was 89% in group 1, 75% in group 2, 78% in group 3 and 42% in group 4. The full dose and partial dose coverage decreased with the income groups of countries, from 70% and 6% in group 1, to 43% and 8% in group 2, to 33% and 11% in group 3, and 20% and 8% in group 4. All 22 EMR countries introduced COVID-19 vaccines by 21 April 2021, but with major inequities in coverage. Additional efforts are needed to address the determinants of unequal vaccine coverage at all stages of the result chain to improve vaccine equity.

A review of EPI progress in Pakistan towards achieving coverage targets: present situation and the way forward.

Pakistan's Expanded Programme on Immunization (EPI) performance has a significant impact on global and regional immunization indicators such as poliomyelitis eradication, maternal and neonatal tetanus and measles elimination. Despite significant efforts by the Government and partners, Pakistan's immunization indicators have not met the expected benchmarks. Barriers to achieving immunization goals are related to limited access to immunization services, lack of parent awareness and weak management. With sustained Government commitment, predictable partner support and by adopting effective strategies, Pakistan can achieve the immunization targets set at the regional and global level and make strong progress towards achieving Millennium Development Goal 4. This paper reviews EPI coverage targets, constraints, costs and resource allocation, and financial impact of suboptimal performance, and indicates the way forward to overcome these challenges.

Review and pooled analysis of studies on MTHFR C677T polymorphism and esophageal cancer.

Esophageal cancer has been associated with tobacco and alcohol consumption, gastric reflux, exposure to nitrosamines from food or other environmental sources, and diets lacking folate. Susceptibility to esophageal cancer may be modified by functional polymorphisms in genes along the folate metabolic pathway, such as methylenetetrahydrofolate reductase (MTHFR). The C677T polymorphism is the most common functional variant, leading to a reduction in enzyme activity. We report a pooled analysis of 5 studies on the association of MTHFR C677T polymorphism and esophageal cancer, including 725 cases and 1531 controls. A significant association between the MTHFR 677 TT genotype and esophageal cancer was observed (OR=2.63, 95% CI: 1.75-3.94), although there was significant heterogeneity between studies. A sensitivity analysis excluded one study; the association between TT genotype and esophageal cancer was still present, although of reduced magnitude (OR=1.57, 95% CI: 0.96-2.56). A significant interaction between smoking and TT genotype on esophageal cancer risk was observed, while no interaction was observed between alcohol consumption and genotype.

Role of Shigella infection in endometriosis: a novel hypothesis.

Endometriosis is the presence of endometrial cells and stroma at ectopic sites outside the uterine cavity. The natural history of endometriosis is uncertain, its etiology unknown, the clinical presentation inconsistent, diagnosis difficult and the treatment poorly standardized. It causes significant morbidity due to pelvic pain and infertility among 15-25% of women during their reproductive age. The benign disease causes peritoneal inflammation, fibrosis, adhesions and ovarian cysts but displays features of malignancy, like neo-vascularization, local invasion and distant metastasis. Mechanical, hormonal, immunological, environmental and genetic factors have been implicated in its etiology but provide inconclusive explanations. Present study was carried out on ectopic and eutopic endometriotic tissue specimens collected during laproscopy/laprotomy from cases of endometriosis. mRNA was isolated from the tissues and converted to cDNA by RT and subsequently subjected to differential display Polymerase Chain Reaction using seven sets of arbitrary primers. A unique band was identified only in the ectopic endometriotic tissue, which was sequenced. BLAST search results revealed sequence homology to shigella bacterial DNA leading us to hypothesize that infection may be playing a role in the etiology of endometriosis. This is the first report implicating the role of bacterial infection in the etiology of endometriosis. Shigella is known to invade the mucosa of the colon through the feco-oral route causing Shigellosis. The pathogenesis of shigellosis involves inflammation, ulceration, haemorrhage, tissue destruction and fibrosis of the colonic mucosa resulting in abdominal pain and diarrhoea/dysentery, this is similar to the pathogenesis of endometriosis which also involves inflammation, haemorrhage, tissue destruction and fibrotic adhesions of the pelvic peritoneum resulting in abdominal pain and infertility. The non-motile shigella bacteria invade the deeper mucosal layers by travelling from cell to cell of colonic epithelium, reaching the lamina propria of the colonic mucosa. We propose that, by the same mechanism, the bacteria travel across the colon wall to reach the outer peritoneal surface of the colon, which is in close proximity to the posterior uterine surface in the Pouch of Douglas, the site which incidentally happens to be the commonest site of early endometriosis. Our hypothesis therefore proposes that shigella or shigella-like organisms may be the trigger for the initiation of immunological changes in the pelvic peritoneum causing endometriosis. Once the endometrial cells are implanted at ectopic sites they are sustained by hormones and angiogenic factors. Hence "Infection hypothesis" provides a novel explanation for the etiopathogenesis of endometriosis.

Interaction of Glucagon G-Protein Coupled Receptor with Known Natural Antidiabetic Compounds: Multiscoring In Silico Approach.

Glucagon receptor (GCGR) is a secretin-like (class B) family of G-protein coupled receptors (GPCRs) in humans that plays an important role in elevating the glucose concentration in blood and has thus become one of the promising therapeutic targets for treatment of type 2 diabetes mellitus. GCGR based inhibitors for the treatment of type 2 diabetes are either glucagon neutralizers or small molecular antagonists. Management of diabetes without any side effects is still a challenge to the medical system, and the search for a new and effective natural GCGR antagonist is an important area for the treatment of type 2 diabetes. In the present study, a number of natural compounds containing antidiabetic properties were selected from the literature and their binding potential against GCGR was determined using molecular docking and other in silico approaches. Among all selected natural compounds, curcumin was found to be the most effective compound against GCGR followed by amorfrutin 1 and 4-hydroxyderricin. These compounds were rescored to confirm the accuracy of binding using another scoring function (x-score). The final conclusions were drawn based on the results obtained from the GOLD and x-score. Further experiments were conducted to identify the atomic level interactions of selected compounds with GCGR.

The alpha1(VIII) and alpha2(VIII) collagen chains form two distinct homotrimeric proteins in vivo.

The short chain collagen variant, type VIII, is considered to be comprised of two distinct gene products, the alpha1 and alpha2 polypeptide chains. However, recent in vitro translation studies suggest that these chains can form homotrimers. We report here data from biochemical, immunohistochemical and molecular biological experiments, which together provide evidence that alpha1 and alpha2 polypeptides of type VIII collagen exist as homotrimers in cells and tissues. High-performance liquid chromatographic separation of type VIII collagen isolated from Descemet's membrane consistently demonstrated equimolar quantities of the two chains (alpha1:alpha2 1. 03+/-0.02 (S.E.M.); n=41). The availability of highly specific antibodies for the two polypeptides has assisted the in vivo characterisation of type VIII collagen. Immunoprecipitation of trimeric type VIII collagen from Descemet's membrane with purified anti-alpha1(VIII) and anti-alpha2(VIII) yielded fractions that contained only the alpha1(VIII) and alpha2(VIII) chains, respectively. Cultured human mesangial cells synthesised both polypeptides, but the alpha1(VIII) chain was found exclusively in the cell pellet, while the media contained only the alpha2(VIII) chain. The RNA from human mesangial cells and cornea showed message for both chains. However, in peritoneal fibroblast and mesothelial cell RNA, only alpha1(VIII) mRNA was detectable, demonstrating that the transcription of these two genes was not always co-ordinated. Immunohistochemistry showed that both polypeptides were present in cornea, optic nerve, aorta and umbilical cord but did not always co-localise. These results indicate the alpha1(VIII) and alpha2(VIII) chains preferentially form pepsin-resistant, homotrimeric molecules and so can exist as two distinct proteins.

Steroid therapy of a proliferating hemangioma: histochemical and molecular changes.

OBJECTIVES: Hemangioma is a primary tumor of the microvasculature in which angiogenesis is initially excessive, followed by regression of the newly formed vessels. Intervention is necessary in up to 20% of cases, high-dose systemic or intralesional steroids being the first-line treatment. As the mechanism of action of steroids is unknown, we undertook an investigation of the cellular and molecular effects of their action. STUDY DESIGN: A unique opportunity to study the effect of steroid treatment was presented when biopsy material was obtained from an infant with an ulcerated proliferating hemangioma before and after intralesional triamcinolone injection, which resulted in an accelerated regression of the lesion. Histochemical quantitation of mast cells, molecular analysis by reverse transcriptase-polymerase chain reaction (RT-PCR) for 7 growth factor transcripts and differential display RT-PCR (DD RT-PCR) were conducted. RESULTS: After steroid therapy, the mast cell number increased (untreated = 2.22 +/-.27 [standard error of the mean ¿SEM¿]; treated = 8.7 +/-.71 [SEM] mast cells per field, respectively; P <.0001; n = 40 fields for each group), and the transcriptional expression of cytokines: platelet-derived growth factor-A and -B; interleukin-6; transforming growth factor-beta1 and -beta3 decreased, while that of basic fibroblast growth factor (bFGF) and vascular endothelial cell growth factor remained unaltered. Elevated urinary bFGF levels noted in cases of proliferating hemangioma, persisted even after steroid treatment. Using DD RT-PCR an amplicon that shared 100% sequence homology with the human mitochondrial cytochrome b gene was detected in the hemangioma biopsy after steroid treatment. CONCLUSIONS: The regression of this hemangioma subsequent to steroid therapy was accompanied by a significant increase in mast cell density, reduced transcription of several cytokines, and an enhanced expression of the mitochondrial cytochrome b gene.

Clusterin/apoJ expression during the development of hemangioma.

Hemangioma is the most common tumor of infancy. This vascular tumor is characterized by an initial rapid proliferation followed by an inevitable regression. The life cycle of hemangioma is divided into proliferative, involuting, and involuted phases. The cellular and molecular mechanisms responsible for controlling the biological behavior of hemangioma are largely unknown. Differential display analysis using mRNA isolated from biopsy specimens representative of the 3 different phases showed increased expression of clusterin/apoJ (clust/apoJ) in the involuting samples. Clust/apoJ is a multifunctional glycoprotein that has been associated with apoptosis. Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry showed that both the transcription and protein expression of clust/apoJ were increased in hemangioma as the tumor progressed from the proliferative to the involuting and involuted phases. This suggests that clust/apoJ is involved in regulating apoptosis during the spontaneous regression of hemangioma. It has been suggested that mast cells (MC) play a role in the regression of hemangioma. The increase in the number and proportion of clust/ apoJ-positive MC with progression of hemangioma, along with the localization of clust/apoJ to MC granules, supports this hypothesis. We suggest that MC may be synthesizing/releasing this apoptotic modulator, leading to the regression of the tumor. Better understanding of the pathogenesis of hemangioma by identification of the relevant factors involved in its regression such as clust/apoJ will result in the development of novel therapies for this condition and tumors that do not undergo spontaneous regression.

Genotoxicity evaluation of individuals working with photocopying machines.

Photocopying machines are a common sight in the cities of India. There is ample evidence showing that the components of toners individually or in the form of a complex mixture are genotoxic. Toxic components of the photocopiers are from their emissions, toners and extremely low frequency electromagnetic fields (ELF-EMFs). In the present study micronucleus test (MNT) on buccal epithelial cells, cytokinesis block micronucleus (CBMN) assay and chromosomal aberration analysis on peripheral blood mononuclear cells was performed on 98 workers occupationally involved in photocopying and 90 age and sex matched controls. The results showed a significant increase in the frequency of MN in buccal epithelial cells and peripheral blood lymphocytes, as well as chromosomal aberrations in the exposed as compared to the control subjects.

Altered mitochondrial cytochrome b gene expression during the regression of hemangioma.

Hemangioma is a primary tumor of microvasculature. Its development typically exhibits a proliferative phase followed by an involuting phase that continues into the involuted phase. Although apoptosis has been reported, the mechanisms regulating the spontaneous regression of hemangioma are largely unknown. The authors recently demonstrated up-regulation of the mitochondrial cytochrome b gene in hemangioma associated with steroid-induced regression. The present study investigated whether a similar change occurred during spontaneous regression. Biopsy material was obtained from 11 patients with hemangiomas at different phases of development. In one of these patients, a biopsy was taken from the proliferative, involuting, and involuted areas of the hemangioma. In another patient, a biopsy was taken before and 5 weeks after the intralesional administration of steroids. From each tissue specimen, RNA was isolated and subjected to reverse transcriptase-polymerase chain reaction analysis by use of specific primers for the human mitochondrial cytochrome b gene. Semiquantitative reverse transcriptase-polymerase chain reaction analysis revealed that the strongest expression of the mitochondrial cytochrome b transcripts was in specimens taken from hemangiomas in the involuting phase compared with those from the proliferative and involuted phases. The authors concluded that mitochondrial cytochrome b is associated with both the spontaneous and the steroid-induced regression of hemangioma, probably by regulating apoptosis.

Platelet derived growth factor-BB induced calcium transients in cultured human peritoneal mesothelial cells.

The peritoneal membrane is used as an artificial dialysis organ for patients with end-stage renal failure during continuous ambulatory peritoneal dialysis (CAPD). Resident peritoneal mast cells may influence the course of the inflammatory process during acute infection or prolonged exposure to dialysate in a paracrine fashion, with the release of preformed mediators, including platelet derived growth factor (PDGF), which may activate human peritoneal mesothelial cells (HPMC) and other peritoneal cells. PDGF receptor interactions have multiple intracellular effects, typified by cell proliferation, protein synthesis, cytoskeletal organization, extracellular matrix turnover, and angiogenesis. PDGF may mediate HPMC inflammatory response by stimulating cytokine release. The presence of transcripts for both PDGF-B polypeptide and PDGF-beta receptor in cultured HPMC was confirmed using a reverse transcriptase-polymerase chain reaction. To investigate the activity of PDGF-beta receptors in HPMC, the authors examined intracellular calcium (Ca2+(i)) mobilization in HPMC in response to PDGF-BB (100 ng/ml), histamine (1.0 mmol/L), and 4-brA23187 (1.0 micromol/L) using the calcium indicator, fura-2. HPMC calcium mobilization in response to PDGF was assessed in Krebs-Ringer saline, with and without added external calcium (Ca2+(ext)). HPMC responded to PDGF with a transient rise in Ca2+(i) (approximately 1.6-fold) that returned to an elevated resting value. In the absence of Ca2+(ext), PDGF produced a Ca2+(i) transient indicative of Ca2+ release from intracellular stores. These results suggest that HPMC have functional PDGF-beta receptors that may bind and respond to PDGF in both an autocrine and paracrine

Mitochondria and tumors: a new perspective.

BACKGROUND: Mitochondrial DNA (Mt DNA) defects have been identified in a variety of Tumors, but the exact role of these defects in the pathogenicity and tumor progression is poorly understood. This study aims at identifying the status of mitochondrial OXPHOS genes in neoplastic transformation and attempts to establish a cause and effect relationship between mitochondrial OXPHOS defects and tumor progression. MATERIALS AND METHODS: Mutational, expression and functional analysis of l2 of the 13 mitochondrial OXPHOS genes has been carried out using PCR, Real-Time PCR and protein modeling in 180 sporadic samples of a heterogeneous group of benign and malignant tumors like that of benign, malignant, matched blood and adjacent normal tissue of breast and benign hemangioma. RESULTS: Mutations were identified in the ND4L, ND6 and COX-II regions of the mitochondrial OXPHOS genes. All the mutations were limited only to the malignant breast tissues. On relative quantification, a compromised expression of OXPHOS genes was identified in all the malignant tissues irrespective of their mutational states. Protein modeling revealed loss of function mutations of ND6 and COX-II proteins. CONCLUSION: This is the first study worldwide wherein a comparative study using different benign and malignant tumors has been carried out to assess the role of Mt DNA defects. Our data reveals mitochondrial dysfunction only in malignant cells and not in their benign counterparts, indicating that the dysfunction may arise after the pro-proliferative pathway has set in. We hypothesize that compromised OXPHOS may be a responsive mechanism of the cell to counter cancers, rather than a mechanism of initiating tumorigenesis.

Genotoxicity in agricultural farmers from Guntur district of South India--A case study.

BACKGROUND: Pesticides play an important role in controlling the pests on agricultural crops and thereby to increase the yield of agricultural produce. Farmers occupationally exposed to pesticides during spraying activities are more prone to genotoxicity than unexposed. AIM: To assess the genotoxicity in farmers, engaged in spraying complex mixture of pesticides in the cultivation of cotton crops. MATERIAL AND METHODS: A total number of 152 male subjects were selected randomly from Guntur district of Andhra Pradesh (AP), South India. The demographic particulars viz., personal habits, duration of exposure to pesticides, types of pesticides used were collected from the study subjects using an interview schedule. Among them 76 subjects were farmers and the remaining individuals served as unexposed or controls. Blood samples from these subjects were collected for assessing the genetic damage by chromosomal aberrations (CAs) test and micronucleus test (MNT). RESULTS: The results of the study indicated that CA was significantly higher with 2.8% in farmers who were exposed to pesticides when compared to unexposed (0.72%). However, there was a minor difference in MN with 0.13% and 0.12% between exposed and unexposed which was not statistically significant (p < 0.05). CONCLUSION: A correlation between CA frequency and exposure to benzene hexachloride (BHC) pesticide residue was observed.

Psoriasis-associated genetic polymorphism in North Indian population in the CCHCR1 gene and in a genomic segment flanking the HLA-C region.

Psoriasis is a common, chronic, recurrent, inflammatory, hyper proliferative disorder of the skin, which has a relatively high prevalence in the general population (0.6-4.8%). Linkage and association analyses in various populations have revealed a major locus for psoriasis susceptibility, PSORS1, at 6p21.3. Association of the disease with human leukocyte antigen (HLA) Cw6, corneodesmosin (CDSN) and the coiled-coil alpha-helical rod protein-1 (CCHCR1) has also been reported. Though the PSORS1 locus accounts for 30-50% of familial psoriasis in various global population groups, yet no studies have been published from the North Indian population. Some of the SNPs in HLA-C and CCHCR1 genes have been reported as markers for disease susceptibility. Therefore in the present study, DNA samples from psoriasis patients from North India were genotyped for polymorphisms in CCHCR1 and HLA-C genes. The allele frequencies were calculated for patients and controls, and were compared for odds ratio and confidence interval values. SNPn.7*22222 (rs12208888), SNPn.7*22333 (rs12216025), SNPn.9*24118 (rs10456057), CCHCR1_386 (rs130065), CCHCR1_404 (rs130076) and CCHCR1_1364 (rs130071) were found to be significant in psoriasis patients. Linkage disequilibrium analysis revealed two haplotypes (rs12208888, rs2844608, rs12216025, rs10456057, rs130065, rs130066, rs130068, rs130269, and rs12208888, rs2844608, rs12216025, rs130076, rs130066, rs130068, rs130269, rs130071) as highly susceptible haplotypes for psoriasis in the cohort studied. Preliminary analysis of the data also suggests the possibilities of ethnic group specific disease related polymorphisms, pending validation in future studies.

Microsatellite instability analysis and its correlation with hMLH1 repair gene hypermethylation status in esophageal pathologies including cancers.

Cancer development is associated with genetic instability. Identification of specific loci altered during carcinogenesis in a particular tissue gives scope for early detection and predicting the progressive nature of the tissue pathology. Instability at microsatellite loci is widely attributed to mismatch repair errors due to epigenetic alterations. Using three dinucleotide markers, D3S1313, D9S171, D17S250 and two mononucleotide markers BAT25, BATRII, we evaluated MSI in 97 cases enrolled for endoscopy of upper GI tract with symptoms of dyspepsia, reflux or dysphagia. We aimed at evaluating markers that reflect instability in esophageal malignancies, examine the prevalence of MSI in cancers and other pathologies of the esophagus, and determine the methylation status of hMLH1 gene in relation to MSI. 42% (21/50) cancers and 15.4%(2/13) precancers exhibited MSI where 85.7% cancers and 50% precancers with MSI, showed a hypermethylated hMLH1 promoter. Increased number of cases with repair gene methylation were seen with increasing severity of the esophageal pathology suggesting epigenetic progression parallels histologic changes. BAT25 and D3S1313 markers exhibited instability frequently and cases with MSI using these markers showed an abnormal hMLH1 promoter. Thus these markers were useful in identifying the mismatch repair phenotype. These two markers may be useful to screen cases for early cancer related changes, after validation on a larger sample.

Role of BRCA1, HSD17B1 and HSD17B2 methylation in breast cancer tissue.

The pattern of altered gene expression due to epigenetic change is of major importance in malignancies. Aberrant DNA methylation is one of the many potential causes for this and is considered to be an early event in the etiology of breast carcinogenesis. The present study assessed the methylation status of three genes relevant in breast cancer (BC): The breast cancer susceptibility gene 1 (BRCA1), 17 beta hydroxy steroid dehydrogenase type 1 (HSD17B1) and type 2 (HSD17B2). Restriction enzyme based Methylation specific PCR (REMS PCR) was carried out in 104 tumor samples from sporadic BC patients and 48 samples of adjacent normal breast tissue. The percentage of tumor samples showing BRCA1, HSD17B1 and HSD17B2 methylation was 20.4%, 83.3% and 31.3%, respectively. Methylation was higher in tumors when compared to adjacent normal breast tissue samples. This suggests that methylation of these three genes plays an important role in BC etiology. Methylation is responsible for gene silencing and since BRCA1 and HSD17B2 were not found to be methylated in the same tissue samples, this suggests that the etiology of > 50% of the tumors could be accounted for by the independent epigenetic silencing of these two genes. BRCA1 and HSD17B2 genes may increase the risk of developing BC via enhanced estradiol activity. It is for the first time that the role of HSD17B gene methylation in BC pathophysiology is being proposed.

Association of progesterone receptor gene polymorphism (PROGINS) with endometriosis, uterine fibroids and breast cancer.

Endometriosis, uterine fibroids and breast cancer are female health disorders associated with a great deal of morbidity. Since all these disorders are hormone responsive, our present study has been carried out to identify the association of 306bp Alu insertion polymorphism in intron 7 of progesterone receptor gene (PROGINS). DNA was isolated from the blood samples of 445 Asian Indian women, which included 100 endometriosis, 80 fibroids and 157 cases of breast cancer along with 108 age matched normal healthy women as controls. PROGINS polymorphism was assessed by PCR followed by agarose gel electrophoresis. Results showed that T2 allele frequency is 5%, 10% and 14.6% in endometriosis, uterine fibroids and breast cancer, as compared to 5.5% in controls. This indicates that PROGINS can be considered as a predisposing risk marker for breast cancer but not for endometriosis and uterine fibroids.

Effects of five commonly used glucocorticoids on haemangioma in vitro.

1. High-dose systemic or intralesional steroids are the first-line pharmacological treatments for haemangioma. However, the mechanism of action of steroids is unknown. Using the in vitro model developed by us, the present study examined some of the effects of five commonly used glucocorticoids on haemangioma biopsies taken from two patients. 2. At 12 micro mol/L, triamcinolone and dexamethasone consistently exhibited capillary growth inhibition, whereas methylprednisolone displayed an inhibitory effect during the first 7 days of culture. At this concentration, inhibition of capillary growth was observed in betamethasone-treated cultures derived from one patient but not in those derived from the other. However, hydrocortisone had a negligible effect on capillary growth. 3. Transcription of various factors considered important for haemangioma development were studied by reverse transcription-polymerase chain reaction. Neither vascular endothelial growth factor nor fibroblast growth factor-2 played a vital role in steroid-induced inhibition of capillary growth. All glucocorticoids induced a marked decrease of interleukin (IL)-6 transcripts. 4. Capillary growth inhibition in cultures treated with all glucocorticoids, except triamcinolone, was associated with an increased transcription of clusterin/apolipoprotein J (clust/apoJ), an apoptotic gene. There was increased transcription of mitochondrial cytochrome (cyt) b in the inhibited cultures resulting from triamcinolone, dexamethasone or methylprednisolone treatment that was associated with capillary growth inhibition, suggesting an important role of mitochondria in glucocorticoid-induced regression of haemangioma. 5. Our results indicate that glucocorticoids may modulate haemangiogenesis via an upregulation of cyt b, clust/apoJ and/or IL-6. The variable effects of different glucocorticoids on one or more of these factors may explain the interindividual variation in the in vivo response of haemangioma to the steroids.