Evaluation of HPV E6/E7 mRNA Detection in Clinically Suspected Cases of Cervical Cancer with Abnormal Cytology: Time to Upgrade the Screening Protocols.

Background Human papillomavirus (HPV) E6/E7 mRNA tests determine the oncogenic activity of the virus and represent a good clinical biomarker for predicting the risk of cervical cancer. So, the present study was conducted to know the role of HPV E6/E7 mRNA as a predictive biomarker for cervical carcinoma. Methodology The present study was conducted on 55 clinical samples of cervical scrapings and biopsy from the clinically suspected cases (based on signs and symptoms) of cervical cancer having abnormal PAP smear. The samples were processed in three steps-(1) HPV DNA detection, (2) HPV E6/E7 mRNA detection, and (3) histopathological analysis. Results Out of a total of 55 patients, 16 (29.09%) were positive for both HPV E6/E7 mRNA and HPV DNA and six were positive for only HPV DNA. So, a total of 22 (40%) patients were positive for HPV DNA. Out of these 22 samples, 10 (45.5%) were of HPV-16, six (27.3%) were of HPV-18, four (18.2%) were of HPV-31, and two (9.1%) were of HPV-45. Out of total 16 patients positive for HPV E6/E7 mRNA, 10 (62.5%) were of genotype 16 and six (37.5%) were of genotype 18. The patients who were found positive for HPV 31 and 45 genotypes did not have E6/E7 mRNA expression. On colposcopic-guided biopsy, among these 16 samples, eight (50%) were diagnosed with invasive squamous cell carcinoma, six (37.5%) with cervical intraepithelial neoplasia grade 3 (CIN3), and two (12.5%) with CIN2. Out of those six patients in whom only HPV DNA was positive, five had normal biopsy findings and one had CIN1. Conclusion The present study suggests that HPV E6/E7 mRNA detection could be more reliable than DNA testing for predicting the risk of progression of HPV-induced cervical lesions to cervical carcinoma and it can be used as a non-invasive tool for triage and patient follow-up.

Comparative study of effect of Withania somnifera as an adjuvant to DOTS in patients of newly diagnosed sputum smear positive pulmonary tuberculosis.

BACKGROUND: Ashwagandha (Withania somnifera Linn.) a rejuvenative herb has long been used as an immunomodulator in Indian subcontinent. As immunity plays an important role in pathogenesis and treatment of tuberculosis (TB), so role of W. somnifera as an adjuvant has been studied on selected parameter. METHOD: A randomized, double-blind placebo-control study was conducted in two groups of 60 newly diagnosed sputum smear positive pulmonary TB patients on Directly Observed Treatment - short course (DOTS) regime. W. somnifera root extract or placebo capsules were given as add-on therapy for duration of 12 weeks. Effects on sputum conversion, Hemoglobin (Hb), body weight, Erythrocyte Sedimentation Rate (ESR), RBC counts, WBC counts, CD4 and CD8 counts, Serum Glutamic-Oxaloacetic Transaminase (SGOT), Serum Glutamic-Pyruvic Transaminase (SGPT), serum uric acid and HRQL (Health Related Quality of Life) Index scores were studied. RESULTS: At the end of 8 weeks, sputum conversion was seen in 86.6% patients in study group and 76.6% in placebo group. At the end of 12 weeks a highly significant increase was seen in both CD4 and CD8 counts in study group. A raised SGOT and SGPT levels (>35IU/L) were observed in 16.6% and 33.3% patients in study group; 43.33% and 53.33% in the placebo group of patients. Elevated serum uric acid levels (>6mg/dl) were observed in 20% and 33.33% in study and placebo group respectively. Average gain in HRQL score was better in patients of study group. CONCLUSION: Use of W. somnifera as an adjuvant in conjunction with anti-TB drugs used as DOTS showed a favorable effect on symptoms and immunological parameters in patients with pulmonary TB.

Coinfection of chikungunya and dengue viruses: A serological study from North Western region of Punjab, India.

INTRODUCTION: Dengue and chikungunya (CHIK) infections appear to be increasing in all parts of India. Aedes aegypti mosquitoes are common vectors for dengue virus (DENV) and CHIK virus (CHIKV). In areas where both viruses cocirculate, they can be transmitted together. There are very few studies discussing the dengue-chik coinfection from Punjab region of India. The present study was undertaken to study the clinical features of dengue-CHIK coinfection and compare with monoinfection. MATERIALS AND METHODS: IgM antibody capture (MAC) ELISA for dengue IgM and CHIK IgM and ELISA for nonstructural protein 1 antigen was performed on serum samples obtained from suspected patients. RESULTS: Out of total 3160 samples from suspected patients for dengue infection, 2178 (68.92%) samples were positive for DENV while CHIK IgM antibodies were positive in 127 patients out of the total suspected 373 cases (34.04%). In addition to this, 283 samples were tested for both viruses, out of which 27 sera were positive (9.54%) for coinfection of dengue and CHIK. The comparison of signs and symptoms showed that the coinfected patients had fever in all cases while rash was seen in only 30% cases. Arthralgia (79%) and thrombocytopenia (77%) was seen in significant number of coinfected cases thus revealing overlapping nature of dengue-CHIK coinfection. CONCLUSION: Increase in the number of Dengue and Chikungunya infections and their cocirculation is an important public health concern which warrants the implementation of strict control measures.

Hepatitis E virus: A leading cause of waterborne viral hepatitis in Northwest Districts of Punjab, India.

BACKGROUND: Acute viral hepatitis (AVH) caused by enterically transmitted hepatitis A virus (HAV) and hepatitis E virus (HEV) poses a major health problem in developing countries such as India. Despite improving sanitation, heath awareness, and socioeconomic conditions, these infections continue to occur both in sporadic as well as in epidemic forms in different parts of India. AIMS: The aim of this study is to determine the total as well as age-specific prevalence rates of HAV and HEV in the outbreaks of waterborne hepatitis in districts surrounding Amritsar region of Punjab. MATERIALS AND METHODS: The study was conducted in the Virology Research and Diagnostic Laboratory, Government Medical College, Amritsar, during the study period of January 2015-March 2016. Samples from suspected outbreaks of AVH occurring in various districts around Amritsar were included as a part of the study. A total of 95 sera were tested for IgM antibody to HEV and HAV using IgM capture ELISA kit. RESULTS: Out of the total 95 samples received, 73 samples (76.84%) were positive for HAV/HEV. Out of the total positive cases, 65 (68.42%) had HEV infection, 2 (2.1%) had HAV, and 6 cases (6.31%) were coinfected with both HAV and HEV. The 21-30 years age group (25 cases) was identified as the most susceptible group for HEV infection. The coinfected subjects presented a wider range of age distribution (1-10 years: 1; 11-20 years: 3; 21-30 years: 1; 31-40 years: 1). Seasonal distribution of data revealed bimodal peaks for HEV infection. CONCLUSION: There should be some surveillance system to regularly monitor the portability of drinking water from time to time to avoid such preventable outbreaks in future.

Significance of coagulase negative Staphylococcus from blood cultures: persisting problems and partial progress in resource constrained settings.

BACKGROUND AND OBJECTIVES: Coagulase negative Staphylococcus (CoNS) is frequently isolated from blood cultures but their significance is difficult to interpret. CoNS bacteria which are often previously dismissed as culture contaminants are attracting greater importance as true pathogens in the past decades. Clinical evaluation of these isolates suggests that although there is a relative increase of CoNS associated bloodstream infections in recent years, the microorganisms still remain the most common contaminants in blood cultures. The objective of this study was to determine the significance of CoNS isolated from blood cultures. MATERIALS AND METHODS: A retrospective study was conducted to evaluate the rate of contamination in blood cultures in a tertiary care hospital. The paired specimens of blood were cultured using conventional culture methods and the isolates of coagulase negative staphylococci were identified by standard methodology. Clinical data, laboratory indices, microbiological parameters and patient characteristics were analyzed. RESULTS: Of 3503 blood samples, CoNS were isolated from blood culture of 307 patients (8.76%). The isolates were reported as true pathogens of bloodstream infections in only 74 out of 307 cases (24.1%). In the vast majority, 212 of 307 (69.0%), they were mere blood culture contaminants and reported as insignificant/contaminant. CONCLUSION: Determining whether a growth in the blood culture is a pathogen or a contaminant is a critical issue and multiple parameters have to be considered before arriving at a conclusion. Ideally, the molecular approach is for the most part a consistent method in determining the significant isolates of CoNS. However, in countries with inadequate resources, species identification and antibiogram tests are recommended when determining significance of these isolates.

Seroprevalence of Common Viral Diseases: A Hospital Based Study from Amritsar, India.

INTRODUCTION: There is a paucity of published data on epidemiology of the viral infections in Punjab, India. The Viral Research and Diagnostic Laboratory (VRDL) located at Government Medical College (GMC), Amritsar conducts the initial diagnosis of common viruses circulating in the region. AIM: To report the seroprevalence of various viral diseases in Amritsar and neighboring districts of Punjab along with the demographic and geographical data of the patients. MATERIALS AND METHODS: Whole blood sera samples from a total of 5781 patients suspected of various viral diseases (chikungunya, dengue, hepatitis A, hepatitis B, hepatitis C, hepatitis E, herpes simplex-1 or herpes simplex-2) were received at VRDL, GMC, Amritsar from January 2015 to April 2016 and a cross sectional study was conducted. The diagnosis was based on serological assays. RESULTS: Out of total 5781 suspected cases, 1790 were diagnosed with antibodies for atleast one of the tested viral diseases. Dengue virus was the highest occurring viral disease (56.77%). Male patients infected with viruses outnumbered female cases, except for hepatitis E and herpes simplex-1. The 21-40 years age group appeared to be the most susceptible age group for nearly all studied viral infections. Among the total nine reported outbreak investigations, largest encompassed 1335 dengue cases from Amritsar district. CONCLUSION: VRDL established in GMC, Amritsar has become instrumental in detection of emerging and re-emerging viral diseases including outbreak investigation of communicable diseases. Timely action with the help of adequate information and capacity to diagnose the viruses might help in saving numerous lives.

Assessment of bacteriological quality of drinking water from various sources in Amritsar district of northern India.

INTRODUCTION: Safe water is a precondition for health and development and is a basic human right, yet it is still denied to hundreds of millions of people throughout the developing world. Water-related diseases caused by insufficient safe water supplies, coupled with poor sanitation and hygiene, cause 3.4 million deaths a year, mostly in children. METHODOLOGY: The present study was conducted on 1,317 drinking water samples from various water sources in Amritsar district in northern India. All the samples were analyzed to assess bacteriological quality of water for presumptive coliform count by the multiple tube test. RESULTS: A total of 42.9% (565/1,317) samples from various sources were found to be unfit for human consumption. Of the total 565 unsatisfactory samples, 253 were from submersible pumps, 197 were from taps of piped supply (domestic/public), 79 were from hand pumps, and 36 were from various other sources A significantly high level of contamination was observed in samples collected from submersible pumps (47.6%) and water tanks (47.3%), as these sources of water are more exposed and liable to contamination. CONCLUSIONS: Despite continuous efforts by the government, civil society, and the international community, over a billion people still do not have access to improved water resources. Bacteriological assessment of all sources of drinking should be planned and conducted on regular basis to prevent waterborne dissemination of diseases.

Comparison of Enzyme-Linked Immunosorbent Assay and Immunochromatography for Rotavirus Detection in Children Below Five Years with Acute Gastroenteritis.

BACKGROUND: Group-A rotaviruses are responsible for 30 to 60% of severe watery diarrhea cases in young children. Timely diagnosis of rotavirus infection helps to determine appropriate treatment and prevents unnecessary use of antibiotics. AIM: To compare Immunochromatography (ICG) with standard ELISA test for diagnosis of and to determine incidence, clinical socio-epidemiological profile and possible risk factors associated with rotavirus infection in children below five years with acute gastroenteritis. SETTINGS AND DESIGN: A prospective study performed from February 2013 to April 2014 in Microbiology and Paediatrics Departments, Government Medical College, Amritsar, Punjab, India. MATERIALS AND METHODS: Hundred stool samples from children below five years diagnosed with acute gastroenteritis were taken and tested by ICG and standard ELISA test. STATISTICAL ANALYSIS: Performed using the SPSS software for Windows, version 17.0. P-values calculated using χ(2) test for categorical variables. A p < 0.05 was considered significant. RESULTS: Maximum cases with ICG showed a sensitivity of 95.24% and specificity of 97.47% when compared to ELISA. Incidence of rotavirus diarrhea was 21% using ELISA and 23% using ICG. With ELISA rotavirus infection was highest in age group 6 months to 24 months (83.3%) and in male (90.47%). The infection was maximum during November to April and presented with triad of diarrhea, vomiting, fever (76.2%). Majority of cases had watery diarrhea in high percentage (90.47%). Severe dehydration (76.19%), respiratory symptoms (38.09%), bottle feeding (52.38%), malnourished children (47.61%), children playing with toys (47.6%) and submersible water pump (61.95%) as a source of drinking water associated with rotavirus infection were found to statistically significant. CONCLUSION: ICG shows a good agreement with ELISA and has the advantage of being a quicker, cost-effective and useful for testing single specimen, convenient, not requiring additional equipment, readily available, simple to perform and easy-to-read results.

Herpes simplex virus 2 : a boon to develop other sexually transmitted infections.

Genital herpes is one of the most common sexually transmitted infections. Though, mostly asymptomatic, it acts as a potential risk factor for acquisition of other sexually transmitted infections (STIs). The present study was undertaken to know sero-prevalence of herpes simplex virus 2 (HSV2), syphilis and HIV among STI clinic attendees and to detect the diagnostic utility of HSV2 IgM antibodies. The study group included 170 individuals attending STI clinic irrespective of their presenting complaints. Their sera were subjected to enzyme-linked immunosorbent assay for detection of HSV2 IgM antibodies. The samples were also screened for HIV and syphilis. Twenty-seven (15.88%) out of 170 persons were found to be seropositive for HSV2 IgM antibodies. Syphilis was detected in 13 individuals (7.65%). Twelve individuals (7.06%) were found to be reactive for anti HIV I antibodies. Ten of the genital herpes patients (37.04%), did not have any complaint of genital ulcer. Eight HSV2 patients (29.63%) had coinfection with HIV I, 6 (22.22%) with syphilis and 2 (07.41%) had co-infection with both HIV and syphilis. A significant proportion of the patients with genital herpes presented without the history of genital ulcers. Thus, its detection and treatment among asymptomatic patients is significant so as to reduce its transmission and other STIs.

Seroprevalence of HIV infection among the patients attending various emergency departments in a tertiary care hospital.

Emergency departments (EDs) receive patients from every background, socioeconomic group and health status. Hence, EDs can play a critical role in offering human immunodeficiency virus (HIV) testing and help in the national strategy of early HIV detection. The present study was conducted on 400 patients attending various EDs after taking Institutional Review Board approval. They were screened for HIV antibodies by three rapid/simple assay tests having different principles/antigens as per the NACO guidelines. Twenty-three (5.75%) of the 400 patients were HIV reactive. Fifteen (65.22%) of the 23 HIV-reactive patients were unaware of their reactive status. Majority of the HIV-reactive (65.22%) patients were from the Medicine emergency followed by Orthopaedics and Surgery (13.04%). Seven (30.43%) had history of fever of more than 1 month duration. Eight (34.78%) of them were later on clinically diagnosed as having various opportunistic infections. Thus, the study emphasizes the need for expansion of routine voluntary HIV counseling and testing to all the patients who come to the ED and practicing universal work precautions by health care workers.

Prevalence of Methicillin-resistant Staphylococcus Aureus (MRSA) in a Tertiary Care Hospital in Northern India.

AIM: The emergence of Methicillin-resistant Staphylococcus aureus (MRSA) has posed a serious therapeutic challenge. We report the prevalence and antibiotic susceptibility pattern of MRSA in the hospitals attached to GMC, Amritsar, Punjab. MATERIALS AND METHODS: The study comprised of 250 coagulase-positive staphylococci (COPS) isolated from a total of 6743 clinical specimens (like pus, blood, urine, high vaginal swab, sputum, etc.) of patients admitted in hospitals attached to Government Medical College, Amritsar from January 2008-February 2009. Routine antibiotic susceptibility testing was performed and interpreted as per standard guidelines. Methicillin resistance was detected using oxacillin and cefoxitin disc diffusion method, oxacillin screen agar method, and minimum inhibitory concentration using broth macrodilution method. RESULTS: A total of 115 (46%) strains were found to be methicillin resistant. Multidrug resistance was observed in 73% MRSA strains. However, no strain was resistant to vancomycin. CONCLUSION: Regular surveillance of hospital-associated infection and monitoring of antibiotic sensitivity pattern is required to reduce MRSA prevalence.

Role of coenzyme Q10 (CoQ10) in cardiac disease, hypertension and Meniere-like syndrome.

Coenzyme Q10 (ubiquinone) is a mitochondrial coenzyme which is essential for the production of ATP. Being at the core of cellular energy processes it assumes importance in cells with high energy requirements like the cardiac cells which are extremely sensitive to CoQ10 deficiency produced by cardiac diseases. CoQ10 has thus a potential role for prevention and treatment of heart ailments by improving cellular bioenergetics. In addition it has an antioxidant, a free radical scavenging and a vasodilator effect which may be helpful in these conditions. It inhibits LDL oxidation and thus the progression of atherosclerosis. It decreases proinflammatory cytokines and decreases blood viscosity which is helpful in patients of heart failure and coronary artery disease. It also improves ischemia and reperfusion injury of coronary revascularisation. Significant improvement has been observed in clinical and hemodynamic parameters and in exercise tolerance in patients given adjunctive CoQ10 in doses from 60 to 200 mg daily in the various trials conducted in patients of heart failure, hypertension, ischemic heart disease and other cardiac illnesses. Recently it has been found to be an independent predictor of mortality in congestive heart failure. It has also been found to be helpful in vertigo and Meniere-like syndrome by improving the immune system. Further research is going on to establish firmly its role in the therapy of cardiovascular diseases.

Effect of Lactobacillus casei as a probiotic on modulation of giardiasis.

The present study was designed with the aim to study the modulation of giardiasis with natural intervention, i.e., Lactobacillus casei--a probiotic, in a murine model. Probiotics are viable microorganisms having a beneficial effect on the prevention and treatment of specific pathological conditions. Various dairy products such as curd, lassi, and kulfi were employed particularly to isolate live organisms (probiotics). Out of five isolated strains only one was found to be similar to a standard strain--Lactobacillus delbrueckii. Experimentally, it was observed that oral feeding of isolated strain A, 7 days prior to inoculation with Giardia lamblia trophozoites, was not as efficient as L. casei in eliminating Giardia infection from BALB/c mice. Interestingly, it was observed that L. casei fed 7 days prior to Giardia infection was more effective and efficient in eliminating the infection from mice. On histological examination, it was observed that probiotic-fed mice had less atrophied villi and infiltrating cells in the small intestine compared to the control. Ultrastructural studies by scanning electron microscopy further confirmed the protection of the mice dosed with L. casei 7 days prior to Giardia infection and also when simultaneously infected with Giardia. It is felt that probiotics, particularly L. casei, modulate Giardia infection by minimizing or preventing the adherence of Giardia trophozoites to the mucosal surface, suggesting that probiotics offer a safe and effective mode to prevent and treat Giardia infection.