Expression of microRNA-155 correlates positively with the expression of Toll-like receptor 7 and modulates hepatitis B virus via C/EBP-ß in hepatocytes.

Effective recognition of viral infection and successive activation of antiviral innate immune responses are vital for host antiviral defence, which largely depends on multiple regulators, including Toll-like receptors (TLRs) and microRNAs. Several early reports suggest that specific TLR-mediated immune responses can control hepatitis B virus (HBV) replication and express differentially with disease outcome. Considering the versatile function of miR-155 in the TLR-mediated innate immune response, we aimed to study the association between miR-155 and TLRs and their subsequent impact on HBV replication using both a HBV-replicating stable cell line (HepG2.2.15) and HBV-infected liver biopsy and serum samples. Our results showed that miR-155 was suppressed during HBV infection and a subsequent positive correlation of miR-155 with TLR7 activation was noted. Further, ectopic expression of miR-155 in vitro reduced HBV load as evidenced from reduced viral DNA, mRNA and subsequently reduced level of secreted viral antigens (HBsAg and HBeAg). Our results further suggested that CCAAT/enhancer-binding protein-ß (C/EBP-ß), a positive regulator of HBV transcription, was inhibited by miR-155. Taken together, our study established a correlation between miR-155 and TLR7 during HBV infection and also demonstrated in vitro that increased miR-155 level could help to reduce HBV viral load by targeting C/EBP-ß.

Occult HBV infection among anti-HBc positive HIV-infected patients in apex referral centre, Eastern India.

BACKGROUND: The prevalence of occult HBV, defined by the presence of HBV DNA in individuals with antibodies to HBV core antigen and with absence of HBV surface antigen, but its clinical significance and virological features in HIV-infected patients is still unclear. AIM: To investigate the prevalence, clinical significance and molecular characterization of occult hepatitis B virus infection in ART-Naive HIV-positive individuals. MATERIAL AND METHODS: Among the 1077 HIV-infected patients with different risk factors for HIV infection, 297 were HBsAg-ve ART-naive, of them 112 was randomly selected for the study. HBV DNA was tested by in-house PCR and quantified by qPCR. Molecular characterization was performed by sequencing the envelope and overlapping polymerase genes. RESULTS: We found the prevalence of occult HBV to be 10.7% among a randomly selected group of HBsAg-ve/antiHBc+ve HIV-infected patients. Overall 33.9% (38 of 112) of the patients were antiHBc positive indicating exposure to HBV infection. HBV DNA was detected in 12/38 (31.5%) antiHBc positive samples and 50% of them had CD4 T cell count < 200 cells/mm(3). HCV coinfection was low (2.7%). No surrogate marker for OBI could be identified. Presence of antiHBs antibodies did not rule out OBI. Liver biopsy in six cases showed varying stages of chronic hepatitis. Several mutations were detected but not the common immune escape mutant G145R. CONCLUSION: In conclusion the prevalence of OBI was significantly high among HIV coinfected patients, which highlights the importance of HBV DNA testing in these patients and indicates need for further prospective studies in larger cohorts to assess its clinical significance.

Subgenotypes of hepatitis B virus genotype D (D1, D2, D3 and D5) in India: differential pattern of mutations, liver injury and occult HBV infection.

Hepatitis B genotype D (HBV/D) is the most widespread genotype and exists as at least five subgenotypes (HBV/D1-D5). However, little is known about the association of virological characteristics with clinical differences among HBV/D subgenotypes. To investigate the virological characteristics of these subgenotypes and their clinical implications, we selected a cohort of 109 genotype D infected individuals from the state of West Bengal, India, including 68 HBsAg positive patients and 41 with occult HBV infection. Among the HBsAg positive subjects 28 had chronic hepatitis B virus infection, 40 were asymptomatic carriers based on clinical examination, liver function test and ultrasonograph results. Overall, HBV/D1 was found in 17%, HBV/D2 in 29%, HBV/D3 in 34% and HBV/D5 in 20% of the cases. HBV/D1 was significantly associated with chronic liver disease (P = 0.01), and in this subgenotype A1896 (PreC mutations) were most common. Although BCP mutations (A/C1753 and T1762/A1764) were found to be frequently associated with HBV/D2 (33% and 33%) and D5 (47% and 59%), no apparent clinical correlation was observed. On the other hand, occult HBV infection was significantly associated with HBV/D3 infection, along with low level of BCP and PreC mutations and several non-synonymous substitutions in the catalytic reverse transcriptase (RT) domain of polymerase gene. Similar nucleotide substitutions in the surface (S) gene region were observed from both northern and eastern Indian HBV/D3 isolates. In conclusion, HBV/D subgenotypes differ in their mutational patterns in the S, polymerase and the BCP/PreC regions that may influence their clinical outcomes.

Influence of early mobilization program on pain, self-reported and performance based functional measures following total knee replacement.

BACKGROUND: Total knee replacement (TKR) is an optimal treatment for persons with severe knee joint pain and disability, who were unsuccessful with conservative management. Early mobilization can be defined as moving out of bed and/or walking quickly after the surgery for reducing the risks allied with bed rest. There is a paucity of studies on effects of early mobilization on a performance-based measure of timed up and go test (TUG), six-minute walk test (SMWT) and a self-reported disease-specific measure of a knee injury and Osteoarthritis outcome score (KOOS) following TKR. METHODS: A prospective pre-post-trial was conducted at Manipal Hospital, Bangalore, India. Participants underwent early (POD '0') mobilization on the same postoperative day within 7 h post-TKR surgery. Outcome measures were recorded by an independent blinded observer. The statistical significance level was set at 'p' value < 0.05. The difference between pre-operative and post-operative outcome measure at 1 month and 3 months post-intervention were analyzed using repeated measures of ANOVA. RESULTS: The study included a total of 78 participants (59 Females; 19 Males) and the mean age of the included participants was 64.1 ±â€¯7 years. Amongst, 78 participants, 53 underwent unilateral TKR, 25 underwent bilateral TKR. There were three dropouts in the study due to post-operative complications. Significant improvements from pre-operative to one month were observed following POD '0' mobilization on NPRS (7.35 ±â€¯1.2 to 4.3 ±â€¯1.7), SMWT (169 ±â€¯70 to 236.7 ±â€¯80.7). KOOS subscales of pain, symptom, and quality of life showed significant changes at one month and 3 months. TUG, Knee strength, Knee ROM and KOOS ADL subscale shown improvements only at 3 months post-intervention. CONCLUSION: Our study findings suggest that POD '0' (early) mobilization can result in reduced pain and an increase in walking speed at 1 month. Significant changes were observed in pain, Knee strength, Knee ROM, TUG, SMWT and KOOS subscales at 3 months following total knee replacement.

Adaptive Sampling for Sound Propagation.

Precomputed sound propagation samples acoustics at discrete scene probe positions to support dynamic listener locations. An offline 3D numerical simulation is performed at each probe and the resulting field is encoded for runtime rendering with dynamic sources. Prior work place probes on a uniform grid, requiring high density to resolve narrow spaces. Our adaptive sampling approach varies probe density based on a novel "local diameter" measure of the space surrounding a given point, evaluated by stochastically tracing paths in the scene. We apply this measure to layout probes so as to smoothly adapt resolution and eliminate undersampling in corners, narrow corridors and stairways, while coarsening appropriately in more open areas. Coupled with a new runtime interpolator based on radial weights over geodesic paths, we achieve smooth acoustic effects that respect scene boundaries as both the source or listener move, unlike existing visibility-based solutions. We consistently demonstrate quality improvement over prior work at fixed cost.

New Delhi metallo-ß-lactamase and extended spectrum ß-lactamases co-producing isolates are high in community-acquired urinary infections in Assam as detected by a novel multiplex polymerase chain reaction assay.

BACKGROUND: The ability of microorganisms to evade antibiotic pressure is challenging in healthcare as patients have little or no drug treatment options. Detection of the prevalence of antibacterial resistance pattern helps towards improved antibiotic policy and empirical treatment. OBJECTIVES: We carried out antibiogram profiling and documented the prevalence and co-prevalence of New Delhi metallo-ß-lactamase (NDM) and extended spectrum ß-lactamases (ESBL) encoding genes in urinary Escherichia coli and Klebsiella pneumonia isolates. MATERIALS AND METHODS: Antibiotic susceptibilities were tested for 241 isolates of E. coli and K. pneumoniae from urine samples collected from out- and hospitalised patients. Polymerase chain reaction (PCR) was carried out on isolates tested positive for phenotypic production of metallo-ß-lactamase and ESBL. A multiplex PCR assay was designed to detect the genes. RESULTS: Multiplex PCR assay designed had a limit of detection of 10 3 CFU/mL in vitro. NDM detected was significantly higher among K. pneumoniae compared to E. coli (69.2% vs. 18.2%; P = 0.001). Of 17, 14 NDM positive isolates also harboured ESBL genes. The co-production of CTX-M + TEM + NDM (3/9; 33.3% and 5/8; 62.5%) was most common in K. pneumoniae and E. coli, respectively while CTX-M + TEM + SHV + NDM was found in one isolate. Of the 156 phenotypically ESBL producing isolates, CTX-M, TEM and SHV was detected by PCR in 85, 53 and 24 isolates, respectively. CONCLUSION: NDM and ESBL co-producing isolates were both community (64.7%) and hospital (35.29%) acquired among E. coli. Antibiotic resistance can be effectively evaluated by a cost and time effective molecular method, such as the multiplex PCR used in this study, which complement culture and sensitivity tests.

Expression of Mts1, a metastasis-associated gene, increases motility but not invasion of a nonmetastatic mouse mammary adenocarcinoma cell line.

The mts1 gene codes for a 101 amino acid protein belonging to the S100 subfamily of Ca(2+)-binding proteins. Mts1 is overexpressed in metastatic cancers as compared to their nonmetastatic counterparts, and although mts1 is known to be involved in the metastatic phenotype (Davies et al., 1993; Grigorian et al., 1993), the role mts1 plays in this process is not clearly understood. In order to determine what role mts1 plays in the process of metastasis, we have performed transfection studies on nonmetastatic and metastatic mouse mammary adenocarcinoma cell lines, CSML0 and CSML100, respectively (Senin et al., 1983, 1984). The metastatic variant, CSML100, expresses high levels of mts1, whereas the nonmetastatic variant, CSML0, expresses almost no mts1. CSML0 cells transfected with mts1 were assessed in in vitro motility and invasion assays, as well as in vivo metastasis assays to determine the role of mts1 in these processes. Cell lines expressing mts1 display an altered morphology as well as increased motility in modified Boyden chemotaxis chambers. However, no significant increase in in vitro invasion or in in vivo metastasis was observed. Therefore, the presence of mts1 may be important for metastasis by increasing motility, but may not be sufficient for invasion in vitro or metastasis in vivo. Very low levels of type IV collagenase activities were observed in CSML0 cells and the transfectants, as opposed to the highly metastatic CSML100 cells, where high levels of type IV collagenase activities were observed. It is possible that the presence of these proteases in addition to mts1 may be responsible for the high metastatic potential of the CSML100 in vivo.

Hepatitis B virus genotype D infection among antenatal patients attending a maternity hospital in Calcutta, India: assessment of infectivity status.

A seroepidemiological study was conducted to determine the hepatitis B virus carrier rate and infectivity status among antenatal patients in Calcutta. Fifteen of 400 antenatal mothers (3.74%) were carriers of HBV genotype D. Four (1.0%) of them were positive for HBeAg with a high risk of transmitting infection to their babies. The presence of precore mutant HBV, which has been associated with transmission events from HBeAg mothers to their babies, was not detected in any of the HBeAg negative mothers.

Hemostatic abnormalities following Echis carinatus (saw-scaled viper) envenomation in the rhesus monkey.

Hemostatic abnormalities following lethal and sublethal doses of Echis carinatus venom were studied in a subhuman primate model. The observations suggest the occurrence of disseminated intravascular coagulation of varying magnitude within a few hours of envenomation. A procoagulant effect of the venom of Echis carinatus is likely to be the major cause of the disturbance in the hemostatic mechanism.

Studies on Escherichia coli as a cause of acute diarrhoea in Calcutta.

The prevalence of different types of diarrhoea-producing Escherichia coli among 240 patients with acute diarrhoea in hospital was investigated. The 25 patients (10.4% of the total) from whose faeces we isolated enteropathogenic E. coli (EPEC) were all less than 5 years old but the 29 (12.1%) from whom we isolated enterotoxigenic E. coli (ETEC) were of various ages, most of them greater than 12 years old. No enteroinvasive E. coli (EIEC) strains were isolated. ETEC strains that produced heat-labile toxin (LT) were encountered more often than those that produced either heat-stable toxin (ST) alone or both LT and ST. The ETEC isolates were distributed among eight different serotypes, the commonest being O148:H28 (38%). Correlations between enterotoxin production, serotype pattern and possession of colonisation factor antigens I and II were observed.