Exaggerated levels of some specific TLRs, cytokines and chemokines in Japanese encephalitis infected BV2 and neuro 2A cell lines associated with worst outcome.

Japanese encephalitis (JE) disease, a viral brain fever is caused by Japanese encephalitis virus (JEV). Despite the availability of effective vaccines against this deadly infection, JE is the leading cause of epidemic viral encephalitis in children in South-east Asia. There is no treatment available for the JE disease which might be due to incomplete understanding of the pathogenesis of JE virus. The JEV infections lead to permanent neurological deficits even in those who survive from the infection. Activated microglia may play a potentially detrimental role by eliciting the expression of pro-inflammatory cytokines such as interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α (TNF-α) influencing the surrounding brain tissue. Microglial activation, proinflammatory cytokine release and leukocytes trafficking are associated following JEV infection in central nervous system (CNS). How the pattern recognition receptors sense the viral nucleic acid and how the microglial and neuronal cells behaves following JEV infection is still unelucidated. There is scarcity of data on the expression levels of toll like receptors (TLRs), cytokines and chemokines in JEV infection in invitro model. To explore the molecular mechanisms of JEV infection of microglial cells and neuronal cells, we studied the expression profile of TLRs, cytokines and chemokines in JEV infected microglial cell line BV2 and Neuronal cell line Neuro 2A. For the present study, we developed the mouse model of encephalitis by intracerebral (IC) injection of JE virus for virus propagation, disease progression and damage study. Our results demonstrate the exaggerated release of some specific TLRs, cytokines and chemokines in invitro cell culture of microglial and Neuro 2A cell line, which are associated with bad outcome in invivo study.

Impaired Autophagy Flux is Associated with Proinflammatory Microglia Activation Following Japanese Encephalitis Virus Infection.

Role of autophagy in Japanese encephalitis viral (JEV) infection is not well known. In the present study, we reported the role of autophagy flux in microglia activation, neurobehavioral function and neuronal death using a mouse model of JEV. Markers for autophagy (LC3-II/I, SQSTM1/P62, phos-Akt, phos-AMPK), and neuronal death (cleaved caspase 12, H2Ax, polyubiquitin) were investigated by western blot at 1, 3 and 7 days post inoculation. Cathepsin D was measured in cerebral cotex of JEV infected mice spectrophotometrically. Microglia activation and pro-inflammatory cytokines (IL1ß, TNF-α, IFNγ, IL6) were measured by immunohistochemistry, western blot and qPCR analysis. In order to determine the neuroinflammatory changes and autophagy mediated neuronal cell death, BV2-microglia and N2a-neuronal cells were used. Autophagy activation marker LC3-II/I and its substrate SQSTM1/P62 were significantly increased while cathepsin D activity was decreased on day 7 post inoculation in cerebral cortex. Microglia in cortex were activated and showed higher expression of proinflammatory mRNA of IL1ß, TNF-α, IFNγ and IL6, with increased DNA damage (H2AX) and neuronal cell death pathways in hippocampus and neurobehavioral dysfunction. Similar observations on JEV infection mediated autophagy flux inhibition and neuronal cell death was found in N2a neuronal cell. Collectively, our study provides evidence on the role of autophagy regulation, microglial activation and neurodegeneration following JEV infection.

Interleukin-10 polymorphisms and susceptibility to ARV associated hepatotoxicity.

Interleukin-10 (IL-10) is an anti-inflammatory cytokine associated with the inhibition of HIV replication. IL-10 polymorphisms were found to be linked to drug-induced hepatotoxicity. Hence we examined the prevalence of IL-10 (-819C/T,-1082A/G) polymorphisms in a total of 165 HIV patients which included 34 patients with hepatotoxicity, 131 without hepatotoxicity and 155 healthy controls by the PCR-RFLP method. In HIV patients with hepatotoxicity, the IL-10-819TT genotype increased the risk of ARV associated hepatotoxicity severity (OR = 1.61, P = 0.35). IL-10-819TT genotype was overrepresented in patients with hepatotoxicity as compared to healthy controls (26.5% vs. 13.5%, OR = 1.61, P = 0.46). IL-10 -819CT genotype was associated with advance HIV disease stage (OR = 0.49, P = 0.045). In HIV patients without hepatotoxicity, the IL-10-819TT genotype was more prevalent in patients consuming tobacco as compared to non-users (OR = 1.60, P = 0.41). In HIV patients without hepatotoxicity using both alcohol + efavirenz along with IL-10 -819CT genotype resulted in increased risk for the acquisition of ARV associated hepatotoxicity (OR = 4.00, P = 0.36). In multivariate logistic regression, taking nevirapine was associated with the risk hepatotoxicity severity (OR = 0.23, P = 0.005). In conclusion, an insignificant association between IL-10 polymorphisms and susceptibility to ARV associated hepatotoxicity.

Real time detection and quantification of Epstein Barr virus in different grades of oral gingivitis and periodontitis patients.

BACKGROUND: Periodontal diseases are of microbial etiology and are globally causing loss of teeth in adult population. Many severe oral diseases have been recently associated to Herpes viruses, of which Epstein Barr Virus (EBV) and human cytomegalovirus (HCMV) have been indicated in the etiology of periodontal diseases. AIM: The purpose of the study was to compare the effect of EBV in different types of periodontal diseases namely acute gingivitis, chronic gingivitis, acute and chronic, localized and generalized aggressive (juvenile) periodontitis and apical periodontitis. MATERIAL AND METHOD: 70 individuals were included in this study. Supragingival plaque and plaque from two deepest sites of the periodontal pockets were collected then stored at 70° c and prepared for nucleic acid extraction. For EBV detection, DNA were extracted from the plaque samples with the QIAamp DNA mini kit. Q-PCR was performed by targeting the non-polymorphic Epstein-Barr nuclear antigen-1 (EBNA-1) gene using Corbett Research 6000 Q-PCR instrument and Rotor gene 6000 software. RESULTS: Overall prevalence of EBV in the disease group was 60% (27/45 patients) as compared to only 8% (4/25 people) in the normal population. The mean copy number of EBV DNA was found to be significantly higher in periodontitis (2234 ± 1811.34) when compared to gingivitis (554 ± 537.64, p = .001) and normal patients (370 ± 161.03, p < .001). CONCLUSION: Here, we found that the prevalence of EBV as well as copy number of EBV was significantly higher in periodontitis patients as compared to gingivitis patients or normal population.

Evaluation of Microscopic observation drug susceptibility (MODS) assay as a rapid, sensitive and inexpensive test for detection of tuberculosis and multidrug resistant tuberculosis.

BACKGROUND: Microscopic observation drug susceptibility (MODS) assay has been suggested as a low cost method for rapid, accurate detection of tuberculosis (TB) and multidrug resistant tuberculosis (MDR-TB). METHODS: A total of 2424 samples collected from 1063 eligible patients of suspected pulmonary or extrapulmonary TB were subjected to MODS assay. Performance of MODS was compared with culture and drug susceptibility testing (DST) by conventional solid Lowenstein-Jensen (LJ) media or liquid Mycobacteria Growth Indicator Tube (MGIT) culture. RESULTS: When compared to reference gold standard of positivity in either solid or liquid reference culture, the MODS assay had sensitivity, specificity, positive predictive value and negative predictive value of 91.3%, 98.2%, 96.0% and 95.9% respectively. MODS took a median time of 10.3 days to culture positivity as compared to 13.8 days using MGIT and 30.5 days using LJ culture. Culture and DST being concurrent in MODS, the median turnaround time for DST was the same as that for culture i.e. 10.3 days. The overall median turn around time for culture positivity and DST using manual MGIT and LJ medium was 23.6 days and 61.2 days respectively. The concordance between MODS culture and the reference susceptibility method was 97.7% for rifampicin, 95.6% for isoniazid, 98.5% for rifampicin and isoniazid. The cost of performing a single MODS assay was INR 200. CONCLUSION: MODS is a rapid and sensitive, yet simple and inexpensive test that may be helpful to enhance diagnostic accuracy, and case detection of TB and MDR-TB in resource constrained settings.

Clinical characterization of influenza A and human respiratory syncytial virus among patients with influenza like illness.

Influenza A and Respiratory Syncytial Virus (RSV) has been recognized as a major cause of acute respiratory tract infection. H1N1 is one of the subtypes of influenza A, pandemic worldwide in July 2009, causing 18,449 deaths globally. To investigate the prevalence and clinical manifestation of the influenza A, H1N1pdm09, and RSV. Throat/nasal swab collected from the patients of all age group either outpatients/inpatients having respiratory illness from 2 to 5 days. The clinical data were recorded in a predesigned questionnaire. RNA was extracted and analyzed by real time PCR at a tertiary care center, 2009-2014. Total 4,352 samples tested for influenza A and H1N1. Out of 4,352, 32.2% (median positivity 21%; range 16-41% during 6 years) were positive for influenza A and 19% were H1N1 (median positivity 16.7%; range 8.7-23% during 6 years). Total 1653 samples were analyzed for RSV from 2011 to 2014, 12% were RSV positive (median positivity 11.35%; range 10-16.3% during 4 years). Pharyngitis, dyspnea were frequent symptoms in influenza A and H1N1 (P < 0.005) whereas bronchiolitis and pneumonia were commonly present in RSV (P < 0.005). The positivity of influenza A and H1N1 was higher in age-group 21-30, whereas RSV in infant and children. H1N1 and RSV were co-circulated and have common clinical symptoms particularly in lower age group. Therefore, laboratory confirmation is necessary for further disease prognosis. Age was an important risk factor that affects the positivity of influenza A, H1N1, and RSV. Different clinical manifestation of H1N1 and RSV will be helpful for early and accurate diagnosis. J. Med. Virol. 89:49-54, 2017. © 2016 Wiley Periodicals, Inc.

Cytomegalovirus infection in pregnant women and its association with bad obstetric outcomes in Northern India.

BACKGROUND: Cytomegalovirus (CMV) infection during pregnancy is far more complex than other infections, due to ability of the virus to be frequently reactivated during the child bearing age and may vertically transmitted to the developing fetus in spite of maternal immunity. Therefore, in the current study we determined the prevalence of CMV infection in pregnant women and tried to identify the role of maternal CMV infection in adverse pregnancy outcomes in Northern India. In this case-control study, 517 pregnant women, out of them 200 in case group and 317 in the control group. The overall 31.72% (164/517) cases were found with active CMV infection. CMV positivity (p=0.026) was significantly associated with bad obstetric history (75/200, 37.50%) compared to normal pregnancy (89/317, 28.07%). CMV infection was predominantly observed in age group 21-25 years. CMV positivity have been found to be significantly higher in women from rural area as compare to those from urban area (p=0.028). However, no significant difference has been observed in case of occupation, income, and haemoglobin level.

Incidence, risk factors and associated mortality of central line-associated bloodstream infections at an intensive care unit in northern India.

OBJECTIVE: To evaluate the incidence, risk factors and associated mortality of central line-associated bloodstream infection (CLABSI) in an adult intensive care unit (ICU) in India. DESIGN: This prospective observational study was conducted over a period of 16 months at a tertiary care referral medical center. SETTING: We conducted this study over a period of 16 months at a tertiary care referral medical center. PARTICIPANTS: All patients with a central venous catheter (CVC) for >48 h admitted to the ICU were enrolled. INTERVENTION AND MAIN OUTCOME MEASURES: Patient characteristics included were underlying disease, sequential organ failure assessment (SOFA), acute physiology and chronic health evaluation (APACHE II) scores and outcome. Statistical analysis of risk factors for their association with mortality was also done. RESULTS: There were 3235 inpatient-days and 2698 catheter-days. About 46 cases of CLABSI were diagnosed during the study period. The overall rate of CLABSI was 17.04 per 1000 catheter-days and 14.21 per 1000 inpatient-days. The median duration of hospitalization was 23.5 days while the median number of days that a CVC was in place was 17.5. The median APACHE II and SOFA scores were 17 and 10, respectively. Klebsiella pneumoniae was the most common organism (n = 22/55, 40%). Immunosuppressed state and duration of central line more than 10 days were significant factors for developing CLABSI. SOFA and APACHE II scores showed a tendency towards significance for mortality. CONCLUSIONS: Our results underscore the need for strict institutional infection control measures. Regular training module for doctors and nurses for catheter insertion and maintenance with a checklist on nurses' chart for site inspection and alerts in all shifts are some measures planned at our center.

Entomological and serological investigation of Japanese encephalitis in endemic area of eastern Uttar Pradesh, India.

BACKGROUND & OBJECTIVES: Japanese encephalitis virus (JEV), a mosquito borne pathogen, is one of the major causes of viral encephalitis in eastern Uttar Pradesh, India. The objective of this work was to evaluate the entomological based virological surveillance of Japanese encephalitis (JE) in the highly endemic area of eastern Uttar Pradesh. METHODS: The study was carried out during September 2010 to March 2013 in Gorakhpur district of Uttar Pradesh. A total of 251 adult mosquito pools and 64 water samples containing larvae were collected from the District of Gorakhpur. Water pH, turbidity, and oxygen level were analyzed for vector breeding index (BI). In addition, 393 serum/cerebrospinal fluid (CSF) samples of acute encephalitis syndrome (AES) suspected cases were collected from the district hospital. RESULTS: The various Culex species found included, Cx. quinquefasciatus (26.83%), Cx. vishnui (22.29%), Cx. pseudovishnui (20.73%), Cx. tritaeniorhynchus (12.71%), Cx. whitmorei (9.04%), and Cx. gelidus (8.25%). Highest minimum infection rate (MIR) was calculated for Cx. tritaeniorhynchus (2.32), followed by Cx. vishnui (1.98) and Cx. pseudovishnui (0.71). All the larvae samples were negative for JEV. The mean number larvae of Cx. tritaeniorhynchus and Cx. pseudovishnui was negatively correlated with pH (r = - 0.45 and r = - 0.63) and turbidity (r = - 0.30 and r = - 0.37). In contrast, positive correlation was observed in case of Cx. quinquefasciatus. A total of 41 clinical samples were found positive for JEV by IgM ELISA. The rainfall was significantly associated with Japanese encephalitis incidence and showed positive correlation to disease transmission (p = 0.02, r = 0. 66). INTERPRETATION & CONCLUSION: The findings showed the rapid dissemination of JEV within a population, facilitated by different species of Culex in the region. As JE is a vaccine-preventable disease, an immunization programme, an effective vector control strategy and application of standard hygiene practices in these endemic areas could result in a considerable reduction in morbidity and mortality due to JE.

Matrix metalloproteinases and their tissue inhibitors in serum and cerebrospinal fluid of children with Japanese encephalitis virus infection.

The expression of matrix metalloproteinases (MMPs) is tightly regulated at the level of gene transcription, conversion of pro-enzyme to active MMPs, and the action of tissue inhibitors of metalloproteinases (TIMPs). The present study aimed to investigate the expression of some specific MMPs (2, 7, 9) and TIMPs (1, 2, 3) in serum and cerebrospinal fluid (CSF) of children with Japanese encephalitis virus (JEV) infection. Serum and CSF levels of MMPs and TIMPs in children with JEV infection and disease control (DC) were compared. The CSF and serum concentrations of MMP-2, TIMP-2 and TIMP-3 were significantly higher in children with JEV infection compared to DC. The concentration of MMP-9 in serum was significantly higher in children with JEV infection than in the DC and healthy control (HC), while in the CSF, no significant difference was observed compared to DC. The MMP-7 serum concentration was significantly higher in children with JEV infection compared to HC, but no significant difference was observed compared to DC. MMP-7 concentration was undetectable in CSF in both groups. The TIMP-1 CSF concentration was significantly higher, while the serum concentration was significantly lower, in children with JEV infection compared to DC. No correlation was found between the levels of each biomolecule measured in CSF and serum, suggesting that the levels in CSF represent local production within the CNS rather than production in the periphery. We also observed leucocytosis, mononuclear pleocytosis and elevated protein concentrations in the CSF of children with JEV infection compared to DC.

Rapid detection of drug resistance and mutational patterns of extensively drug-resistant strains by a novel GenoType® MTBDRsl assay.

BACKGROUND: The emergence of extensively drug-resistant tuberculosis (XDR-TB) is a major concern in the India. The burden of XDR-TB is increasing due to inadequate monitoring, lack of proper diagnosis, and treatment. The GenoType ® Mycobacterium tuberculosis drug resistance second line (MTBDRsl) assay is a novel line probe assay used for the rapid detection of mutational patterns conferring resistance to XDR-TB. AIM: The aim of this study was to study the rapid detection of drug resistance and mutational patterns of the XDR-TB by a novel GenoType ® MTBDRsl assay. MATERIALS AND METHODS: We evaluated 98 multidrug-resistant (MDR) M. tuberculosis isolates for second line drugs susceptibility testing by 1% proportion method (BacT/ALERT 3D system) and GenoType ® MTBDRsl assay for rapid detection of conferring drug resistance to XDR-TB. RESULTS: A total of seven (17.4%) were identified as XDR-TB by using standard phenotypic method. The concordance between phenotypic and GenoType ® MTBDRsl assay was 91.7-100% for different antibiotics. The sensitivity and specificity of the MTBDRsl assay were 100% and 100% for aminoglycosides; 100% and 100% for fluoroquinolones; 91.7% and 100% for ethambutol. The most frequent mutations and patterns were gyrA MUT1 (A90V) in seven (41.2%) and gyrA + WT1-3 + MUT1 in four (23.5%); rrs MUT1 (A1401G) in 11 (64.7%), and rrs WT1-2 + MUT1 in eight (47.1%); and embB MUT1B (M306V) in 11 (64.7%) strains. CONCLUSIONS: These data suggest that the GenoType ® MTBDRsl assay is rapid, novel test for detection of resistance to second line anti-tubercular drugs. This assay provides additional information about the frequency and mutational patterns responsible for XDR-TB resistance.

Upregulated expression of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases in BALB/c mouse brain challenged with Japanese encephalitis virus.

BACKGROUND: Uncontrolled immune responses in the nervous system are potentially damaging following Japanese encephalitis virus (JEV) infection. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) act together to control the proteolysis of extracellular matrix. Disbalances in the MMP/TIMP system during virally induced neurodegenerative processes and inflammations are responsive to changes in the progression of diseases. METHODS: The expression of MMP-2, MMP-7, MMP-9, TIMP-1, and TIMP-3 in JEV-infected mouse brain was analyzed by RT-PCR for semiquantitation and ELISA for estimation of protein along with brain histopathology at different days postinoculation (dpi). Gelatin gel zymography was performed for MMP-2 and MMP-9 activities. RESULTS: In the virus-infected group, expression of MMP-2, MMP-7, MMP-9, TIMP-1, and TIMP-3 was found to be increased from 1 dpi to 6 dpi as compared to controls by both RT-PCR and ELISA. The expressions of MMPs and TIMPs at mRNA and protein levels were in concordance with each other. Post hoc multiple comparison analysis between days revealed that, in the virus-infected groups, significant increases (p < 0.05) in MMP and TIMP levels were observed between various dpi at both mRNA and protein levels. Only the MMP-7 protein level at 6 dpi was not significant compared to 5 dpi (p = 0.99). CONCLUSION: Overexpression of MMPs and TIMPs is associated with disease severity in the central nervous system (CNS) during JEV infection. Our results showed that JEV infection can alter the expression of MMPs and TIMPs in the CNS. Thus, assessing these important immune mediators in CNS infection appears to play an important role in the development of symptoms and may help to understand the JEV-induced neurological disorders. More studies are required on this important enzymatic system to study their role in immune mediated pathogenesis.

Epidemiology and genetic relatedness of measles virus infection in Uttar Pradesh, India, during 2009-2010.

Measles surveillance and epidemiological investigation are beneficial tools for genetic analysis and documentation of the interruption of transmission of endemic measles. In this study, areas of Uttar Pradesh, India, associated with measles outbreaks were investigated. Random blood and urine samples were collected from clinically defined cases. The cases were investigated through extensive house-to house surveys. The cases were diagnosed clinically and serologically, and through genotyping of the virus. All of the cases were found to be serologically positive for measles. In the studied population, a 1.9% case fatality rate and an overall 16% attack rate of measles virus were found. Out of 10 outbreak areas, the measles virus D8 genotype was found in nine, and the D8 and A genotypes were found in the remaining area. This study calls for an improved surveillance system and intensive characterization of genotypes in circulation for the measles elimination program in India.

Trends of anti-tuberculosis drug resistance pattern in new cases and previously treated cases of extrapulmonary tuberculosis cases in referral hospitals in northern India.

BACKGROUND: Drug-resistant tuberculosis is one of major current challenges to global public health. The transmission of resistant strains is increasing as a burden of multidrug-resistant tuberculosis (MDR-TB) patients in extra pulmonary tuberculosis (EPTB) cases in India. AIM AND OBJECTIVES: The aim was to study trends of anti-tuberculosis drug resistance pattern in new cases and previously treated cases of EPTB in referral hospitals in northern India. STUDY DESIGN AND SETTING: A prospectively observational study and referral medical institutions in northern India. MATERIALS AND METHODS: All EPTB specimens were processed for Ziehl Neelsen staining, BACTEC culture and BACTEC NAP test for Mycobacterium tuberculosis complex. All M. tuberculosis complex isolates were performed for radiometric-based drug susceptibility pattern against streptomycin, isoniazid, rifampicin and ethambutol using the 1% proportion method. RESULTS: We found that 165/756 (20.5%) isolates were identified as M. tuberculosis complex by the NAP test. We observed that 39.9% were resistant to first-line antitubercular drugs. The resistance rate was higher in previously treated patients: H (30.3%), R (16.3%), E (15.7%) and S (16.3%). MDR-TB was observed in 13.4%, but, in new cases, this was 11.4% and 19.1% of the previously treated patients (P<0.05). CONCLUSION: MDR-TB is gradually increased in EPTB cases and predominant resistance to previous treated cases of EPTB. The molecular drug sensitivity test (DST) method can be an early decision for chemotherapy in MDR-TB patients. The International Standards of TB Care need to be used by the RNTCP and professional medical associations as a tool to improve TB care in the country.

Mannose-binding lectin (+54) exon-1 gene polymorphism influence human immunodeficiency virus-1 susceptibility in North Indians.

Mannose-binding lectin (MBL) is a circulating pattern-recognition molecule involved in the innate immune system that mediates phagocytosis and activates complement by binding to carbohydrate motifs. MBL-2 allelic variants are associated with deficiencies in innate immunity and have been found to be correlated with human immunodeficiency virus (HIV) infection. The present study investigated the role of MBL-2 exon-1 gene polymorphism (A, B, C and D) in 180 HIV-1 seropositive (HSP) stratified on the basis of disease severity (stage I, II, III), 50 HIV-1 exposed seronegative (HES) and 305 HIV-1 seronegative (HSN) individuals as a possible factor in the susceptibility to HIV-1 infection and the influence on disease progression among North Indian individuals. In our population, gene frequencies of MBL-2 variants were 15%, 5% and 2% for B, C and D alleles, respectively. The frequency of A/O heterozygous genotype was higher (42.00%), mainly because of A/D in HES group compared with HSP (35.00%) and HSN (36.72%) group. Homozygous B/B genotype was more frequent in HSP (6.11%) group than in HSN (1.31%; P = 0.005; odds ratio (OR) = 4.898) and was significantly associated with fourfold risk of acquiring HIV-1 infection. Our findings indicate that homozygosity for the codon 54-allele associated with low MBL production in the exon-1 of the MBL-2 gene is associated with increased susceptibility to HIV-1 infection in the studied population.

Incidence of ESBL producers amongst Gram-negative bacilli isolated from intra-abdominal infections across India (based on SMART study, 2007 data).

OBJECTIVES: This study was conducted in 9 centers spread over India from January 1 to December 31, 2007 to monitor in vitro susceptibility of Gram-negative bacilli to Group I carbapenem, ertapenem and other antimicrobials in intra-abdominal infections and to identify early changes in susceptibility pattern of community or hospital acquired organisms, with a focus on ESBL producers. MATERIAL AND METHODS: Gram-negative bacilli isolated from intra-abdominal samples of patients with documented intra-abdominal infections were processed for identification by conventional/ automated methods and antimicrobial susceptibility by Micro-Scan (Siemens) MIC panel against 12 antimicrobials (3rd and 4th generation cephalosporins, Groups I and II carbapenems, amikacin, levofloxacin, amoxicillin-clavulanic acid and piperacillin-tazobactam). RESULTS: A total of 588 isolates were identified, of which 351 (60%) were E. coli and 114 (19%) were Klebsiella spp. 79% of E. coli and 70% of Klebsiella spp. were ESBL producers in general. 110 of E. coli and 35 of Klebsiella isolates were from community-acquired intra-abdominal infections. 80% of E. coli and 63% of Klebsiella isolates from community-acquired infections were ESBL producers, against 79% of E. coli and 73% of Klebsiella isolates from hospital-acquired infections. Amongst the ESBL-positive isolates of E. coli, 94% were susceptible in vitro to ertapenem, 96% to imipenem and 76% to piperacillin-tazobactam. For ESBL-positive isolates of Klebsiella spp., the corresponding figures were 80%, 94% and 59% respectively. CONCLUSION: The study showed a high incidence of ESBL-producers amongst Enterobacteriaceae isolates from intra-abdominal infections in both community-acquired and hospital-acquired settings across India. Ertapenem was comparable with imipenem against ESBL-positive E. coli isolates, while imipenem was more effective than ertapenem against ESBL-positive Klebsiella isolates.

Successful medical management of renal zygomycosis: a summary of two cases and a review of the Indian literature.

We present two cases of renal zygomycosis caused by Apophysomyces elegans and Mycocladus corymbifer in previously healthy immunocompetent males and an overview of the disease in India. In both cases a percutaneous nephrostomy (PCN) was performed and the etiologic agents were identified by direct microscopy and culture. Amphotericin B was administered and both patients recovered completely. A review of the literature revealed 42 cases of renal zygomycosis in India. The majority of them were from the Postgraduate Institute of Medical Education and Research, Chandigarh, in North India. In contrast to cases from the developed world where transplant recipients and patients with hematological malignancies seem to be most vulnerable to zygomycosis, the most common risk factor in India is uncontrolled diabetes mellitus. However, renal zygomycosis is an exception and the patients in both of our cases had no identifiable underlying disorder and recovered successfully without nephrectomy. It is important to emphasize that treatment of A. elegans must be aggressive and lipid formulations of antifungals are typically favored due to their limited side effects profile and ability of the clinician to use higher doses. A high index of clinical suspicion and knowledge of the varied manifestations in diagnosing this condition cannot be overemphasized.

Variation in the host ABO blood group may be associated with susceptibility to hepatitis C virus infection.

This study aimed to determine the relationship between hepatitis C virus (HCV) infection and ABO/Rhesus blood groups, age and sex. A total of 20 000 patients who came to donate blood in the blood bank of GSVM Medical College, Kanpur were enrolled in the study. Demographic data recorded for each patient included age, sex and blood group. Blood samples were tested for anti-HCV antibodies and ABO/Rhesus blood group antigen typing was performed. The overall positive rate of anti-HCV was 0.34%. We found that seropositivity for HCV increased with age. Anti-HCV antibodies were detected in 1/765 women (0.13%), compared to 67/19 235 men (0.35%). Seroprevalence of HCV was found to be higher in blood group O individuals (0.42%) and lowest in blood group AB individuals (0.04%). The results of this study demonstrate that that HCV infection may not be related to age and sex but the possible association of blood group antigens with HCV infection cannot be ruled out.

Analysis of influenza virus-induced perturbation in autophagic flux and its modulation during Vitamin D3 mediated anti-apoptotic signaling.

Vitamin D3/Calcitriol supplementation in humans is associated with reduced incidence and severity during influenza A virus (IAV) infection. Apoptosis in response to IAV infection is a major contributor to host cell death and tissue damage; however, its modulation by Vitamin D3 remains unclear. In this study, we demonstrate the efficacy of Vitamin D3 in preventing apoptosis induction by pandemic influenza A (H1N1)pdm09 virus in human alveolar cells (A549). Human alveolar epithelial cell line A549 was used to assess the cytotoxic effects of IAV infection. Immunoblotting and fluorescence microscopy were used to study apoptosis and autophagy. The results of the present study demonstrate that IAV induces apoptosis by subversion of host autophagy via down-regulating components of autophagic machinery involved in autophagosome-lysosome fusion and lysosomal activity. Vitamin D3 restores the autophagic flux inhibited by IAV by upregulating the expression of Syntaxin-17 (STX17) and V-type proton ATPase subunit (ATP6V0A2) thereby causing a concomitant decrease in cellular apoptosis via a Vitamin D3 receptor (VDR) dependent mechanism. The present study suggests that Vitamin D3 is a potentially useful agent for limiting IAV-induced cellular injury via its pro-autophagic action.

A single-nucleotide polymorphism in TLR4 is linked with the risk of HIV-1 infection.

INTRODUCTION: Toll-like receptors (TLRs) are pattern recognition receptors that play a role in innate immunity. Mounting evidence shows that single-nucleotide polymorphisms (SNPs) in TLRs link to various infectious diseases, including human immunodeficiency virus (HIV). We hypothesized links between two TLR4 SNPs (rs4986790 leading to Asp299Gly and rs4986791 leading to Thr399Ile) and HIV, to investigate the frequency of TLR4 polymorphism and its role in patients infected with HIV. MATERIALS AND METHODS: We recruited 160 HIV-1 seropositive patients, who were further divided on disease severity based on CD4 count (stages I, II and III), and 270 age- and sex matched healthy HIV-1 seronegative individuals. Subjects were genotyped for TLR4 gene polymorphism by polymerase chain reaction restriction fragment length polymorphism. RESULTS: The TLR4 Asp299Gly heterozygous genotype (OR=2.160; p=0.004) and the mutant allele G (OR=2.051; p=0.002) was higher in HIV-1 infection than healthy controls and also in stage I (OR=2.559; p=0.034) compared to different clinical stages of infection. There was no link between the Thr399Ile polymorphism and HIV infection. CONCLUSION: The TLR4 (Asp299Gly) SNP is a risk factor in HIV-1 disease susceptibility.