Perianal Tuberculosis: Lessons Learned in 57 Patients From 743 Samples of Histopathology and Polymerase Chain Reaction and a Systematic Review of Literature.

BACKGROUND: Few data are published on perianal tuberculosis. OBJECTIVE: This study aimed to determine the best method to diagnose tuberculosis in patients with fistula-in-ano and to conduct a systematic review to determine the incidence and characteristics of tuberculosis fistula-in-ano. DATA SOURCES: The prospective study data and existing literature were derived from PubMed, Google scholar, and Scopus STUDY SELECTION:: Prospective analysis of patients with tuberculous fistula-in-ano treated between 2014 and 2018 was conducted, and a systematic review of studies describing ≥3 patients with tuberculosis fistula-in-ano was completed. INTERVENTION: Testing of tuberculosis was performed by histopathology or polymerase chain reaction of tissue or pus from the fistula tract. MAIN OUTCOME MEASURES: The primary outcomes measured were the detection rate of various tests to detect tuberculosis in fistula-in-ano and the prevalence rate of tuberculosis in simple versus complex fistulas. RESULTS: In 637 samples (410 patients) tested, tuberculosis was detected in 49 samples (43 patients). Additional samples (n = 106) sent in patients with a high index of suspicion tested positive in 14 more patients. Thus, overall, 63 samples tested positive in 57 patients (total: 743 samples in 410 patients were tested). Tuberculosis was detected in 2 of 181 patients (1.1%) in tissue (histopathology), in 28 of 341 patients (8.2%) in tissue (polymerase chain reaction), and in 19 of 115 patients (16.5%) in pus (polymerase chain reaction) samples. To detect tuberculosis, tissue (polymerase chain reaction) was significantly better than tissue (histopathology) (28/341 vs 2/181, p < 0.00001) and pus (polymerase chain reaction) was significantly better than tissue (polymerase chain reaction) (19/115 vs 28/341, p < 0.0009). Tuberculosis was significantly more common in complex fistulas than in simple fistulas (20.3% vs 7.2%, p = 0.0002). The systematic review (n = 199) highlighted that tubercular fistulas are more common in recurrent and complex fistulas and in tuberculosis endemic regions. LIMITATIONS: The true sensitivity and specificity of each testing modality could not be determined because not all patients with tuberculosis fistula-in-ano were tested by every diagnostic modality studied. CONCLUSIONS: The tuberculosis detection rate of polymerase chain reaction was significantly higher than histopathology. Among polymerase chain reaction, pus had higher detection rate than tissue. Tuberculosis was associated with more complex and recurrent fistulas.

Serum Uric Acid Level among Acute Stroke Patients.

Stroke is one of the leading causes of death and disability in developed as well as developing countries like Bangladesh. Elevated serum uric acid levels may predict an increased risk for cerebro-vascular (CV) events including stroke. Aim of the study was to measure the serum uric acid level among stroke patients and determine the relationship between serum uric acid level and stroke. This descriptive, cross-sectional study was carried out in Department of Medicine, Mymensingh Medical College Hospital, Mymensingh, Bangladesh to measure serum uric acid level among 102 stroke patients in a period of one year by using non-probability sampling procedure. Finally, collected data were analyzed using SPSS software Version 17.0. It was observed that the mean age of patients was 60.87±8.05 years, of them 80(78.43%) patients were male and the rest 22(21.57%) were female. About 66(64.70%) of respondents were in age group 60 years and above, while 36(35.30%) were in age group 59 years and below. At least 23(22.55%) of stroke patients had elevated serum uric acid with a mean serum uric acid level of 5.18mg/dl and standard deviation 1.26mg/dl. About 23(27.38%) patients in ischemic stroke had elevated serum uric acid whereas 18(100%) patients in hemorrhagic stroke had normal uric acid level. Uric acid level was elevated in ischemic stroke than haemorrhagic stroke patients (p<0.001). High uric acid level may be considered as a risk factor in patients with acute ischemic stroke.

Genetic variant in the BCL11A (rs1427407), but not HBS1-MYB (rs6934903) loci associate with fetal hemoglobin levels in Indian sickle cell disease patients.

India along with Nigeria and DRC contribute to 57% of the world sickle cell anemia population. The annual number of newborns in India with SCA was estimated at 44,000 in 2010. Even with this high prevalence there is minimal information about genetic factors that influence the disease course in Indian patients. The current study was conducted on 240 patients with SCD and 60 with sickle cell trait, to determine the association of genetic variants at the BCL11A (rs1427407) and HBS1-MYB (rs6934903) loci with fetal hemoglobin levels (HbF). Both these loci have been implicated with influencing HbF levels, a powerful modulator of the clinical and hematologic features of SCD. Our results indicate the BCL11A rs1427407 G>T variant to be significantly associated with HbF levels {19.12±6.61 (GG), 20.27±6.92 (GT) and 24.83±2.92 (TT) respectively} contributing to ~23% of the trait variance. Interestingly no association of the HBS1L-MYB rs6934903 with the HbF levels was seen. The present study indicates the BCL11A (rs1427407) but not HMIP (rs6934903) to be associated with elevated HbF levels in Indian patient. Further interrogation of additional variants at both the loci; as also a GWAS which may help uncover new loci controlling HbF levels.

Promoter variants in interleukin-6 and tumor necrosis factor alpha and risk of coronary artery disease in a population from Western India.

INTRODUCTION: A central component of the atherosclerotic process is inflammation. Single nucleotide polymorphisms (SNPs) present in the promoter region of various cytokines can lead to altered levels of the transcript and a state of low-grade inflammation exacerbating the risk of coronary artery disease (CAD). The present work tries to understand the role of permissive promoter variants in the interleukin-6 gene (IL-6-174G/C) and the tumor necrosis factor alpha (TNFα-308G/A) in the causation of CAD and also dyslipidemia. MATERIALS AND METHODS: Genotyping was conducted on 100 cases of CAD and 150 controls by the allele termination assay SNaPshot. Biochemical parameters were determined by routine enzymatic endpoint methods. The results were analyzed by appropriate statistical methods. RESULTS: No differences in the minor allele frequency IL-6-174G/C SNP were seen between cases and controls (0.13 vs. 0.12). The differences in the allele frequency of TNFα-308A between cases (6%) and controls (2%) have led to an odds ratio, 3.370; 95% confidence interval, 1.039-11.543; P=0.033 in the univariate analysis. In the final logistic regression analysis, however none of the variants were associated with an increased risk of CAD. CONCLUSIONS: In summary, no association of the permissive promoter variants in the IL-6 gene and the TNFα gene were seen with an increased CAD risk. These and other studies highlight the importance of doing population specific studies.

Study of Human Brucellosis among Patients with Pyrexia of Unknown Origin by Antibody Detection.

This study was performed to determine the seropositivity of human brucellosis among the patients suffering from pyrexia of unidentified origin. This cross-sectional study was performed at department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh from September 2018 to August 2019; among the patients of pyrexia of unknown origin visited inpatient and outpatient facility of department of Medicine and department of Paediatrics, Mymensingh Medical College Hospital (MMCH) in Mymensingh division of Bangladesh. A total of 400 serum samples were screened by Brucella-specific latex agglutination test to determine seropositivity. Seven percent (7.0%) (28/400) serum samples were found to be seropositive for brucellosis by detecting Brucella-specific antibody at a titer ≥1:160. Therefore, Brucella-specific latex agglutination test may be recommended as a screening test for human brucellosis in developing and underdeveloped countries.

Distribution and Pattern of Anti-Tubercular Drug Resistance in Patients with Pulmonary Tuberculosis in Mymensingh Region of Bangladesh.

Globally, the emergence of multidrug-resistant strains of Mycobacterium tuberculosis is an increasing problem that adversely affects patient care and public health. This cross sectional descriptive study was carried out in the Department of Microbiology, Mymensingh Medical College from January 2010 to December 2010 to isolate M. tuberculosis from smear-positive sputum samples by Lowenstein-Jensen (L-J) media and investigate the drug resistance pattern. Among 101 smear-positive cases 80(79.20%) yielded growth of Mycobacteria, 5(4.95%) were contaminated and 16(15.84%) showed no growth. Among 80 isolates 76(95.0%) were M. tuberculosis and the remaining 4(5.0%) were Non-tuberculous Mycobacteria (NTM). Out of 76 M. tuberculosis 27(35.52%) were resistant to at least one drug, 4(5.26%) to Isoniazid (INH), 1(1.32%) to Rifampicin (RMP), 8(10.53%) to Streptomycin (SM) and 0(0.0%) to Ethambutol (EMB) and multi-drug resistant tuberculosis (MDR-TB) was 9(11.84%). The present study creates the impression that fairly high rate of anti-tuberculosis drug resistance among the tuberculosis cases and also high MDR-TB (Resistant to both Rifampicin and Isoniazide). The emergence of MDR-TB poses significant trouble to TB control activities throughout the world. The complexity of MDR-TB operation makes it essential to produce new skills to design, plan, application and monitor interventions for the management of MDR-TB. More surveillance and immediate remedial interventions should be performed to combat the trouble of MDR-TB to the general population.

POEMS Syndrome.

A 49 years old male patient admitted with 2 years history of lower extremity symmetrical sensorimotor polyneuropathy, sclerodermic skin change, erectile dysfunction, hepatosplenomegaly and monoclonal gammopathy. The clinical evaluation met the criteria for the diagnosis of (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, skin changes) POEMS syndrome. The patient was treated with corticosteroid and melphelan and responded well. We present a case different from the other cases with severe unusual burning sensation all over the body, which was his sole complaint and with this complaint he visited lot of doctors including psychiatrist.

TP53 codon 72 polymorphism and cervical cancer: a pooled analysis of individual data from 49 studies.

BACKGROUND: Cervical cancer is caused primarily by human papillomaviruses (HPV). The polymorphism rs1042522 at codon 72 of the TP53 tumour-suppressor gene has been investigated as a genetic cofactor. More than 80 studies were done between 1998 and 2006, after it was initially reported that women who are homozygous for the arginine allele had a risk for cervical cancer seven times higher than women who were heterozygous for the allele. However, results have been inconsistent. Here we analyse pooled data from 49 studies to determine whether there is an association between TP53 codon 72 polymorphism and cervical cancer. METHODS: Individual data on 7946 cases and 7888 controls from 49 different studies worldwide were reanalysed. Odds ratios (OR) were estimated using logistic regression, stratifying by study and ethnic origin. Subgroup analyses were done for infection with HPV, ethnic origin, Hardy-Weinberg equilibrium, study quality, and the material used to determine TP53 genotype. FINDINGS: The pooled estimates (OR) for invasive cervical cancer were 1.22 (95% CI 1.08-1.39) for arginine homozygotes compared with heterozygotes, and 1.13 (0.94-1.35) for arginine homozygotes versus proline homozygotes. Subgroup analyses showed significant excess risks only in studies where controls were not in Hardy-Weinberg equilibrium (1.71 [1.21-2.42] for arginine homozygotes compared with heterozygotes), in non-epidemiological studies (1.35 [1.15-1.58] for arginine homozygotes compared with heterozygotes), and in studies where TP53 genotype was determined from tumour tissue (1.39 [1.13-1.73] for arginine homozygotes compared with heterozygotes). Null results were noted in studies with sound epidemiological design and conduct (1.06 [0.87-1.29] for arginine homozygotes compared with heterozygotes), and studies in which TP53 genotype was determined from white blood cells (1.06 [0.87-1.29] for arginine homozygotes compared with heterozygotes). INTERPRETATION: Subgroup analyses indicated that excess risks were most likely not due to clinical or biological factors, but to errors in study methods. No association was found between cervical cancer and TP53 codon 72 polymorphism when the analysis was restricted to methodologically sound studies. FUNDING: German Research Foundation (DFG).

Status of TMPRSS2-ERG fusion in prostate cancer patients from India: correlation with clinico-pathological details and TMPRSS2 Met160Val polymorphism.

BACKGROUND: Prostate cancer (PCa) shows considerable clinical heterogeneity that has been primarily attributed to variable molecular alterations. TMPRSS2-ERG fusion is one such molecular subtype that has been associated with predominantly poor prognosis. More recently, a single nucleotide polymorphism (SNP) in the TMPRSS2 gene rs12329760 C>T (Met160Val) has been shown to positively correlate with the fusion status and also to be associated with increased risk for PCa. The aim of the present study is to determine the frequency of TMPRSS2-ERG fusion and association of rs12329760 in Indian PCa patients with fusion status. METHODS: TMPRSS2-ERG fusion by fluorescence in situ hybridization was determined in 102 of 150 PCa biopsy-proven cases. Genotyping for rs12329760 was performed on the entire cohort of 150 cases by Sanger sequencing. RESULTS: TMPRSS2-ERG fusion was seen in 27 of 102 (26%) cases. Fusion-positive patterns in this study showed fusion by translocation in nine of 27 cases (33.5%), by deletion in six of 27 (22%) cases, and by insertion in 12 of 27 cases (44.5%). No association of the fusion status with Gleason Score, pattern, or perineural invasion was seen. The TMPRSS2 SNP rs12329760 'T' allele was prevalent with a frequency of 0.27 in the PCa patients. The SNP was significantly associated with fusion [odds ratio (OR) = 2.176, 95% confidence interval (CI) = 1.012-4.684, P = 0.04], more specifically fusion by deletion (P = 0.04). CONCLUSION: The results provided here determine the frequency of TMPRSS2-ERG fusions (26%) in a fairly large cohort of Indian PCa cases and also the association of rs12329760 SNP with TMPRSS2-ERG fusion. No association with other clinico-pathological features was observed. Future studies with clinical outcomes are warranted in this population.

The Prevalence of Extended-Spectrum Beta-Lactamase-Producing Multidrug-Resistant Escherichia Coli in Poultry Chickens and Variation According to Farming Practices in Punjab, India.

BACKGROUND: Agricultural use of antimicrobials in subtherapeutic concentrations is increasing in response to the rising demand for food animal products worldwide. In India, the use of antimicrobials in food animal production is unregulated. Research suggests that many clinically important antimicrobials are used indiscriminately. This is the largest study to date in India that surveys poultry production to test for antimicrobial resistance and the occurrence of extended-spectrum ß-lactamases (ESBLs) modulated by farming and managerial practices. OBJECTIVES: Our goal was to survey poultry production for resistance to eleven clinically relevant antimicrobials and phenotypic occurrence of ESBLs as modulated by farming and managerial practices. METHODS: Eighteen poultry farms from Punjab were surveyed, and 1,556 Escherichia coli isolates from 530 birds were tested for susceptibility to 11 antimicrobials using the disk diffusion method and validated using VITEK 2 (bioMérieux, Marcy-L'Étoile, France). Samples from 510 of these birds were phenotypically tested for ESBL production using the combination disk method and confirmed using VITEK 2. Generalized linear mixed models were used to infer differences in resistance profiles associated with different farming practices and facility types. RESULTS: Resistance profiles were significantly different between broiler and layer farms. Broiler farms were 2.2 [ampicillin (AMP), p=0.017] to 23 [nalidixic acid (NX), p<0.001] times more likely to harbor resistant E. coli strains than layer farms. Adjusting for farm type (broiler vs. layer), the odds of resistance (although not statistically significant) to all antimicrobials except nitrofurantoin (NIT) were higher in independent facilities (IUs) as compared to contracted facilities (CFs). Increased prevalence of multidrug resistance (MDR; 94% compared to 60% in layers), including prevalence of ESBL-producing strains (87% compared to 42% in layers), was observed in broiler farms. CONCLUSIONS: Our findings suggest that unregulated use of clinically relevant antimicrobials in Indian broiler and layer farms may contribute to the emergence of resistance and support the need to curb the nontherapeutic use of medically important antimicrobials in food animal production. https://doi.org/10.1289/EHP292.

Curcumin, a potential inhibitor of MMP-2 in human laryngeal squamous carcinoma cells HEp2.

UNLABELLED: Curcumin (diferuloylmethane) has been widely studied for its tumor inhibiting and anticarcino-genic properties. In the present communication, we studied the effect of curcumin on matrix-metalloproteinase-2 (MMP-2), integrin receptors, and focal adhesion kinase (FAK) in human laryngeal cancer cells, HEp2. METHODS: HEp2 cells were treated with curcumin (5 microM) for 30 days and then grown without curcumin for 28 days. Effect of curcumin on MMP-2 expression and activity and on membrane type matrixmetalloproteinase-1 (MT1-MMP), FAK, and integrin receptors was studied by zymography, Western blot, ELISA, RT-PCR, and cell adhesion assay. RESULTS: Treatment of HEp2 cells with curcumin downregulated MMP-2 expression and activity and expression of integrin receptors, FAK, and MT1-MMP to almost background levels. MMP-2 (but not MMP-9) mRNA expression was abolished on curcumin treatment, indicating specific inhibition of MMP-2. Invasive potential of HEp2 cells was also significantly reduced. After drug withdrawal, expression of MMP-2, integrin receptors, MT1-MMP, and FAK returned to control levels. However, MMP-2 activity in serum free medium remained low. CONCLUSIONS: Downregulation of integrin receptors and low levels of FAK may hinder integrin-mediated signal transduction, preventing upregulation of MMP-2 activity. Reduction of MMP-2 activity and inhibition of HEp2 cell invasion by curcumin strongly indicate the potential of curcumin as an inhibitor of tumor cell invasion and metastasis.

Modifications and evaluation of double antibody radioimmunoassay of human carcinoembryonic antigen.

Double antibody radioimmunoassay of carcinoembryonic antigen (CEA), a cancer-associated antigen of the human digestive system, was subjected to certain modifications and critically evaluated. Modifications pertained to: (a) the production of a high titer goat anti-CEA antiserum that was rendered highly specific by solid phase immunoabsorption with cyanogen bromide-activated Sepharose conjugates of normal plasma liver, and colon perchloric acid-soluble glycoprotein antigens: (b) the introduction of suitable alterations in the experimental conditions of radioiodination procedure to minimize and to prevent breakdown of the antigen, thus prolonging the storage of the labeled antigen; (c) the extended incubation period of CEA-anti-CEA immune reaction; and (d) the use of sodium acetate buffer, pH 6.1. Furthermore, the use of an automatic pipetting station for accurate and rapid reagent dispensation and statistical analysis of the radioimmunoassay data on a modern computer to ensure strict quality control of the assay provided some definite improvement over the existing assay.

Differential binding of nuclear proteins to the TP53 gene promoter in male breast tumour.

It is well established that TP53 regulates the expression of many genes, but the regulation of expression of TP53 itself is poorly understood. Recently, it has been shown that there is a tissue-specific binding of nuclear proteins in the TP53 gene promoter. The aim of this study was to determine the nuclear proteins that bind to the TP53 promoter elements (between -104 and -458) in male breast cancer. The results of our study, using the electrophoretic mobility shift assay (EMSA) and Southwestern analysis, have showed: (1) nuclear proteins or factors other than p53 bind to the TP53 promoter; (2) the levels of at least four nuclear proteins vary between normal and tumour breast tissue; and (3) two newly discovered nuclear proteins bind to the TP53 promoter in tumour tissue but are absent in normal tissue. This differential binding of nuclear proteins to the TP53 gene promoter might play a critical role in TP53 transcription and cancer progression in male breast tumours.

Increased ADP-ribosylation of histones in oral cancer.

The involvement of ADP-ribosylation has been investigated by comparing the extent of ADP-ribosylation in two different stages of oral cancer with that of the corresponding normal oral tissue. Poly(ADP-ribose) synthetase(PADPRS), the enzyme that is responsible for this type of posttranslational covalent modification, was assayed first by measuring 14C-ADP-ribose incorporation into different acceptor proteins of the tissue homogenate. The results clearly indicate an increase in PADPRS activity during oral carcinogenesis. After observing this increased pattern in total tissue, further experiments were conducted to check the extent of ADP-ribosylation in purified nuclei. The data of this experiment also indicates a progressive increase in the extent of ADP-ribosylation with increase in malignancy of oral tissue. Further analysis of an ADP-ribosylation of chromosomal proteins indicates that the increased pattern of ADP-ribosylation is mainly attributed to histones and not to non-histone chromosomal proteins. Our investigation suggests that ADP-ribosylation may play a role in oral cancer and may be used as a potential tumour marker.

Differential expression of c-jun and c-myc in N-nitroso diethylamine-induced hepatic oncogenesis in AKR mice.

Expression of c-jun, c-myc, c-fos and c-Ha-ras was examined and correlated with the various stages of N-nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis in male AKR mice. The treated groups were given NDEA 100 ppm, in drinking water for 120 days. The histopathology along with the expression of oncogenes were studied at different durations of treatment such as after 1 day, 3 days, 6 days, 9 days, 15 days, 20 days 30 days, 60 days, 90 days and 120 days of treatment. The results of histological investigation indicate mild hyperplasia and anisonucleosis at 30 days of treatment and prominent pathological features from 60 days onwards until the appearance of hepatocarcinoma at 120 days even without the development of any preneoplastic or neoplastic nodule. The results of the Northern blot hybridization clearly indicate an increased expression of c-jun from 15 days onwards. This overexpression of c-jun at such an early stage indicates its association with the events earlier than the neoplastic changes. However, the persistent overexpression of c-jun at all durations of treatment indicates its association with the events during the later stage of hepatocarcinogenesis, whereas c-myc overexpression starts from 30 days of treatment and persists until the end of the experiment, i.e. 120 days of treatment. However, the results of densitometric quantification indicate that the extent of increase expression of c-myc is less than that of c-jun expression until 1 month of treatment, after which the induction of c-myc exceeds the expression of c-jun. Thus, the overexpression of c-myc from 2 months onwards might be playing a critical role in maintenance of the malignant phenotype. On the other hand, the expressions of c-fos and c-Ha-ras do not have any alteration during NDEA treatment. Thus, our data demonstrate the involvement of c-jun and c-myc in NDEA-induced hepatocarcinogenesis in AKR mice.

Induction of lung carcinogenesis in AKR-mice by N-nitrosodiethylamine/phenobarbitone, associated with high expression of c-myc and c-jun oncoproteins.

Lung carcinogenesis was induced in AKR mice using N-nitrosodiethylamine (NDEA). Tumors were detected in 46.8% of mice provided with 100 ppm NDEA in drinking water. The incidence of tumors was increased to 64.2% when the same carcinogenesis was promoted by phenobarbitone (PB). Lung tumor bearing mice showed no tumors in other organs. Characteristic features of these lung tumors are: (i) appearance of tumors within a short period of time i.e. less than 75 days; (ii) no increase in the number and size of tumors with the increase in dose and duration of treatment of carcinogen; (iii) the same histological type was maintained in more than 80% of tumors. Animals that received treatment for 75-125 days showed no significant advancement in the stage of carcinogenesis in comparison to the 50-75 days treatment period. Moreover, mice which received treatment for 125-150 days, did not have any neoplastic lesions in lungs, but they consisted of liver tumors generally. Expression of oncoproteins, c-myc and c-jun, was detected in all lung tumors but the expression of c-myc protein was more than that of c-jun and both of these oncoproteins were enhanced by the promoter, PB. Highest level of expression of c-myc and c-jun was detected within the period of 50-75 days, whereafter it was decreased significantly within the period of 75-125 days and 125-150 days of treatment. Thus, the results indicate that c-myc/c-jun might be involved in the development of lung cancer in AKR mice, but may not have any role in the maintenance of the malignant phenotype of lungs.

d-limonene chemoprevention of hepatocarcinogenesis in AKR mice: inhibition of c-jun and c-myc.

We have studied the chemopreventive effect of d-limonene, a monoterpene monocyclic compound, against N-nitrosodiethylamine (NDEA) alone and along with phenobarbital (PB) induced hepatocarcinogenesis in AKR mice. Histopathological analysis clearly indicates the maintenance of normal features when the mice were given limonene 15 days prior to carcinogen treatment. The immunohistochemical analysis of c-jun and c-myc oncoprotein shows an increased protein expression (2-3 fold) in NDEA and NDEA-PB mediated hepatocarcinogenesis after 60 days of NDEA treatment. Our earlier work by northern blot analysis has already indicated an increased transcription of c-jun and c-myc in NDEA mediated hepatocarcinogenesis. However, such overexpression of c-myc and c-jun both at mRNA and oncoprotein levels has been completely inhibited when d-limonene was used along with NDEA or NDEA-PB. Thus, the present investigation explains the anti-tumour effect of d-limonene for the first time on the level of oncogene expression in NDEA and NDEA-PB mediated hepatocarcinogenesis.

Chemopreventive effects of green tea polyphenols correlate with reversible induction of p57 expression.

Green tea polyphenols are known to induce apoptosis in certain types of tumor cells. However, the mechanism(s) that enables normal cells to evade the apoptotic effect is still not understood. In this study, Western blot analysis combined with cycloheximide treatment was used to examine the effects of green tea polyphenols on the expression levels of p57, a cyclin-dependent kinase and apoptosis inhibitor, in normal human keratinocytes and in the oral carcinoma cell lines SCC25 and OSC2. The results showed that the most potent green tea polyphenol, (-)-epigallocatechin-3-gallate (EGCG), induced p57 in normal keratinocytes in a dosage- and time-dependent manner, while the levels of p57 protein in oral carcinoma cells were unaltered. The differential response in p57 induction was consistent with the apoptosis status detected by annexin V assay. The data suggest that the chemopreventive effects of green tea polyphenols may involve p57-mediated cell cycle regulation in normal epithelial cells.

Comparative evaluation of indirect and sandwich ELISA for the detection of antibodies to bovine respiratory syncytial virus (BRSV) in dairy cattle.

Seroprevalence of bovine respiratory syncytial virus (BRSV) infection in both exotic and crossbred cattle were described. A baculovirus expressed recombinant purified nucleocapsid (N) protein was used in indirect and sandwich ELISA for screening of 499 bovine sera samples from all over the state for the presence of BRSV antibodies. The seroprevalence rate of BRSV was found to be 46.09% through indirect ELISA while it would found to be 65.33% by sandwich ELISA. The result also indicated that exotic breeds were more susceptible to BRSV infection compared to crossbred cattle. A comprehensive analysis on susceptibility to BRSV as regards to various factors like age and sex was also summarized.

p53 gene mutation in relation to p53 protein accumulation in male and female breast cancer.

The aim of this investigation was to study the prevalence of p53 gene mutations in male and female breast cancers and to find out the relationship between this event and p53 protein expression. Genomic p53 was amplified by polymerase chain reaction (PCR). Exons 5-8 were screened for mutations using single stranded conformation polymorphism (SSCP) analysis. P53 protein expression was detected by immunohistochemistry (IHC) with the monoclonal antibody DO-1. In female breast cancer, p53 gene mutation was detected in 33% cases in either exon 5 or 6. However, in male, mutation was detected only in exon 6 in 90% cases. On the other hand, p53 protein expression was observed in all of these cases. Moreover, p53 protein immunostaining was observed in some of those cancer tissues, where no mutation was detected in exons 5-8. P53 dysfunction, as indicated by mutation or increased protein expression, common in both male and female breast cancer, but rate of occurrence or site of mutation differ from each other. Our results in male breast tumors indicate a positive correlation between p53 mutation and p53 protein overexpression, whereas the results in female breast tumors indicate an overexpression of p53 protein even without p53 gene mutation. Therefore, it may be presumed that p53 protein accumulation can result primarily from mutation. In addition, stabilization of p53 through binding to other proteins is another possible reason of p53 overexpression.