Delay in diagnosis among pulmonary tuberculosis patients of Rayagada District, Odisha, India

Background: India accounts for more than one-fifth of the world's tuberculosis (TB) burden. In spite of efforts taken by the Revised National Tuberculosis Control Programme, tribal areas of the state of Odisha report a high TB incidence over the years. One of the reasons could be delay in reporting to health facilities by the symptomatic patients. During such delays an active case may infect numerous susceptible people thereby contributing to the perpetuation of the infection. The delay in diagnosis may be as long as 2–3 months and even more in hard-to-reach areas. Objective The present study aims to find out the extent of delay in diagnosis among pulmonary TB patients of a tribal dominated district that may help in planning effective control strategies for similar situations. Methods: The information on delay in diagnosis is part of a cross-sectional drug resistance study carried out from June 2011 to May, 2013 in 20 Designated Microscopy Centres (DMCs) of Rayagada district of Odisha, India. Out of 634 smear positive pulmonary TB patients enrolled in this study, information on health seeking by the patients were available for 580 patients. The patients included had clinical signs and symptoms suggestive of pulmonary TB (cough, chest pain, and hemoptysis), with/without radiological evidence. Patients found smear positive by Ziehl–Neelsen microscopy were requested to take part in the study and accordingly a written questionnaire including history of: TB treatment; symptoms experienced by the patient and duration of suffering; and radiological examination was completed. The delay in diagnosis at the DMCs due to delay in health seeking by the symptomatic TB patients was evident as only 5.2% patients reported within 2 weeks and 62.6% up to 1 month of onset of symptoms. Results: The delay in health seeking by the patients was not differentiated by sex or resistance profile, although more men attended the DMC for diagnosis. The present study is the first of its kind to report diagnostic delay of TB among smear-positive TB patients of Rayagada, a tribal-dominated district of Odisha, India and it reveals an extremely long diagnostic delay of TB in this area. We found that 12.9% of patients had a delay exceeding 2 months, and 50% of them had high sputum grade. This is a serious concern due to the fact that each of these patients dispenses up to 3500 bacilli in each cough, and may infect 10–15 people each year, eventually creating a public health problem. Conclusion: Poor awareness of patients about the disease and limited access to health care are the bottom-line in apparent diagnostic delay of TB patients. This substantial patient delay to diagnosis is a major contributing factor for increasing transmission of TB in tribal districts of Odisha. Increased awareness of the disease is crucial in improving health-seeking behavior in these areas

Detection of bacterial and viral pathogens in hospitalized children with acute respiratory illness and determination of different socio demographic factors as important cause of the disease in Odisha, India

The paper an attempt has been made to analysis the status of acute respiratory tract infection among children in India. In the present study we aimed to present first time the detection of viruses, bacteria and mix infection of viruses and bacteria in hospitalized children with ARI and also to analyze the influence of socioeconomic status of parent in two divergent geographical settings of Odisha. Hospitalized children with ARI aged <5 were recruited from July 2014 to June 2015. Nasopharyngeal/Oropharyngial swabs were collected for detection of common respiratory viruses by reverse transcriptase chain reaction (RT-PCR). Bacteria were isolated by routine culture methods. Bivitiate analysis including chi square was used as test of significance. The analysis revealed 150 (56%) were detected with ≥1 bacteria, 40 (15%) with ≥ 1virus, 22 (8.2%) with ≥ 2 bacteria and 20 (7-4%) with both bacteria and virus. Most frequently detected pathogens were Klebsiella pneumonae (18.3%), Sptrptococcus pneumonae (12.7%), Parainfluenza A (36.6%) and Influenza- A18 (30%). Incidences of pathogens were detected more among children <1 year, Gender discrimination in the form of dietary neglect of the female children has also been noted mostly in case of tribal patients. The present study had identified low socioeconomic status, poor housing conditions, illiterate mothers, birth weight, tobacco smoking families and nutritional status as important determinants for ARI. Interventions to improve these modifiable risk factors can significantly reduce the ARI burden among children especially in tribal population

Screening of novel inhibitors targeting Human Papillomavirus 16 E6/AP/P53 ternary complex towards development of therapeutic strategies against HPV-mediated oncogenesis

Cervical cancer is the fourth most common cancer in women worldwide. It is well known that high-risk HPV is the main etiological agent for this infectious viral carcinoma. Human papillomaviruses are small (50 nm) double-stranded DNA viruses composed of a genome of 8 kilobase pair, enclosed inside a non-enveloped capsid protein. The genome includes three portions: (a) early genes (E1, E2, E4, E5, E6, E7) those regulate the vegetative and productive phase of viral life cycle; (b) late genes (L1, L2) which encode the capsid protein and (c) a noncoding regulatory region called long control region (LCR) involved in the regulation of viral replication and transcription. The HPV oncoproteins E6 and E7 recognize numerous host proteins, in large part by hijacking cellular domain-motif interaction networks. E6 and E7 oncoproteins disrupt cell cycle checkpoint control by inhibiting CDKs inhibitors (P21, P27) and degrading P53. In the process of E6 mediated degradation, E6 binds to a short leucine (L)-rich LxxLL consensus sequence within the cellular ubiquitin ligase E6AP3. Subsequently, the E6/E6AP heterodimer recruits and degrades p53. The LxxLL peptide of E6AP is sufficient to render E6 liable to interact with p53 ‘core’ (DNA binding) domain of p53 required for the interaction with E6/E6AP9–11. In the present study, we explored specific novel inhibitors targeting three different druggable pocket i.e., E6-binding cleft, LxxLL pocket of AP and the p53-binding cleft of E6/E6AP/p53 ternary complex using AutoDock tool. A total of five novel compounds with higher binding energy were identified as potential competitive inhibitors against HPV16 E6/AP/P53 ternary complex. The combinatorial strategies targeting these druggable pockets are expected to open up better avenues for the development of therapeutic strategies against HPV-mediated oncogenesis in near future

Molecular mechanisms of HPV mediated neoplastic progression

Abstract Human Papillomavirus is the major etiological agent in the development of cervical cancer but not a sufficient cause. Despite significant research, the underlying mechanisms of progression from a low-grade squamous intraepithelial lesion to high grade squamous intraepithelial lesion are yet to be understood. Deregulation of viral gene expression and host genomic instability play a central role in virus-mediated carcinogenesis. Key events such as viral integration and epigenetic modifications may lead to the deregulation of viral and host gene expression. This review has summarized the available literature to describe the possible mechanism and role of viral integration in mediating carcinogenesis. HPV integration begins with DNA damage or double strand break induced either by oxidative stress or HPV proteins and the subsequent steps are driven by the DNA damage responses. Inflammation and oxidative stress could be considered as cofactors in stimulating viral integration and deregulation of cellular and viral oncogenes during the progression of cervical carcinoma. All these events together with the host and viral genetic and epigenetic modifications in neoplastic progression have also been reviewed which may be relevant in identifying a new preventive therapeutic strategy. In the absence of therapeutic intervention for HPV-infected individuals, future research focus should be directed towards preventing and reversing of HPV integration. DNA damage response, knocking out integrated HPV sequences, siRNA approach, modulating the selection mechanism of cells harboring integrated genomes and epigenetic modifiers are the possible therapeutic targets

HPV Genotypes distribution in Indian women with and without cervical carcinoma: Implication for HPV vaccination program in Odisha, Eastern India

Abstract Background Considering the limited cross protection offered by the current HPV vaccines, understanding the HPV genotype distribution among the different population is essential in predicting the efficacy of current vaccine and devising new vaccine strategy. The present work aimed at investigating the HPV genotypes distribution among women with and without cervical carcinoma in Odisha, Eastern India. Methods A total of 607 participants have been enrolled between January 2014 and June 2016. L1-PCR, sequencing, and E6/E7 nested multiplex type- specific PCR were performed for HPV detection and genotyping. Cytological distribution of 440 cases includes invasive cervical carcinoma or ICC ( n \u2009=\u2009210), inflammatory smear ( n \u2009=\u2009162), normal cytology ( n \u2009=\u200968). Statistical analyses were performed by using SPSS version 20.0 software and MediCal version 14.10.2(7). A p -value of\u2009\u2264\u20090.05 was considered statistically significant. Results The overall prevalence of HPV infection was (359/595) 60.33%. Prevalence of HPV infection was 93.80% (197/210) in invasive cervical cancer (ICC) cases, 54.32% (88/162) in inflammatory smear and 19.11% (13/68) in normal cervical cytology. The most prevalent genotype was HPV16 (87.28%) followed by HPV18 (24.56%) and HPV 51(3.46%). The overall prevalence of single type was 76.58% and highest (78.9%) among ICC cases. The most frequent genotype combination after HPV16\u2009+\u200918(9.4%) was HPV16\u2009+\u200966\u2009+\u200968(2.7%) which was frequently observed in inflammatory cytology. Age\u2009>\u200945years, parity \u22653, low socio-economic condition, rural residential area and post menopause state were significantly associated with HPV infection. Multiple infections did not have a significant association with any of the clinicopathological variables (stage, LN metastasis, cell type) except tumor size\u2009\u2265\u20092cm in ICC cases. The impact of 2v, 4v, and 9v vaccines in preventing cervical cancer in Odisha were 89.99, 91.65, and 92.16% respectively. Conclusion This data would help planning an appropriate strategy for disease monitoring and provides baseline data for post-vaccination surveillance in the region. The nonavalent vaccine would be significant in preventing cervical carcinoma in Odisha. Hence an effective vaccination program based on regional HPV epidemiological profile along with the cervical cancer screening is necessary to reduce the cervical cancer burden in India

HPV genotypes co-infections associated with cervical carcinoma: Special focus on phylogenetically related and non-vaccine targeted genotypes.

HPV is the major causative agent for cervical cancer. Study on the risk of cervical cancer associated with different hr-HPV genotypes would be useful for disease management and new vaccine strategy. With limited reports available, the present study aimed to investigate the pattern of HPV genotypes coinfections and risk of cervical carcinoma associated with them in Indian population. 15 HPV genotypes were detected by E6/E7 multiplex nested type-specific PCR in the HPV-positive cervical samples of 172 cervical cancer cases and 174 subjects with normal cytology. Association between the genotypes and cervical cancer was estimated by calculating the Odds ratio and 95% confidence interval. Risk of cervical carcinoma was associated with multiple genotypes excluding HPV16 (OR:5.87; 95% CI-1.28-26-29; p = .02), multiple genotypes excluding HPV18 (OR = 2.5; 95% CI = 1.09-6.05; p = .03), multiple genotypes of α9 species(OR = 5.3 95% CI = 1.14-24.03; p = .007), and multiple genotypes of α7 species (OR = 2.5; 95% CI = .49-13.45; p = .2). Genotypes not targeted by quadrivalent vaccine types (OR = 2.94 95% CI = 1.48-5.80; p = .001) conferred 2.94 fold higher risk of cervical carcinoma. Cases those coinfected with phylogenetically related genotypes (OR = 2.29; 95% CI(.69-7.59) p = .17) were at 2.9 fold higher risk of invasive cervical carcinoma than those infected with other genotypes although it is not statistically significant. Whereas phylogenetically unrelated genotypes coinfection is negatively associated with cervical carcinoma (OR = .44 95% CI (.244-.8) p = .007) and it is statistically significant.Genotypes not targeted by 9-valent vaccines (OR = .40; 95% CI = .19-.85; p = .017) associated with lesser risk of cervical carcinoma as compared to other genotypes. Subjects infected with any HPV genotype/genotypes excluding HPV16 in association with HPV 18 (OR = 4.1; 95% CI = 1.81-9.25 P = < .001) were at 4.1 fold higher risk of developing invasive cervical carcinoma.In conclusion, the risk of development of cervical cancer is genotype specific and might be associated with type-specific interactions between the genotypes in multiple infections

DETECTION OF JAPANESE ENCEPHALITIS VIRUS AND HERPES SIMPLEX VIRUS IN AES CASES BY ELISA AND PCR IN A TERTIARY CARE HOSPITAL OF EASTERN INDIA

Encephalitis is defined as inflammation of the brain parenchyma associated with neurologic dysfunction. AES occurs in explosive epidemics or in a non-epidemic (sporadic) form. Epidemic in India have been attributed to Japanese encephalitis virus (JEV) infection. Herpes simplex virus (HSV) is responsible for most sporadic cases of AES. This study was undertaken to detect JEV and HSV from CSF samples. Method: Two to three ml of CSF was collected in a dry sterile container. Samples were divided into 2 vials and kept at -20oC. One vial was used for ELISA test in while the other vial was used for PCR. Result: Of 90 cases, 61 were males (68%), 29 were females (32%). Majority belonged to 1 month-5years in 34 (38%) cases followed by 6-10 years in 23 (26%) cases. The commonest symptom was fever in 90 (100%) cases followed by change in mental status in 86 (95.6%).Of 90 samples 2 (2.2%) was positive for JEV in PCR while only 1 (1.1%) was positive in ELISA. A total of 9 (10%) samples were positive for HSV 1 &amp; 2, of which 8 (8.9%) samples were positive in ELISA assay and 8 (8.9%) were positive in PCR assay. In our study only 2 (2.2%) samples were positive for JEV in PCR while one was negative in ELISA. 9 (10%) samples were tested positive for HSV which signifies the sporadic nature of the virus in this region. Conclusion: PCR was found to be more sensitive in detection of JEV, while in HSV detection both ELISA and PCR were equally sensitive. Key words:&nbsp;Japanese B encephalitis virus; Herpes simplex virus; Acute encephalitis syndrome; Polymerase chain reaction; Enzyme linked immunosorbent assay