Liver preservation with machine perfusion and a newly developed cell-free oxygen carrier solution under subnormothermic conditions.

We describe a new preservation modality combining machine perfusion (MP) at subnormothermic conditions(21 °C) with a new hemoglobin-based oxygen carrier (HBOC) solution. MP (n=6) was compared to cold static preservation (CSP; n=6) in porcine orthotopic liver transplants after 9 h of cold ischemia and 5-day follow-up. Recipients' peripheral blood, serial liver biopsies, preservation solutions and bile specimens were collected before, during and after liver preservation. Clinical laboratorial and histological analyses were performed in addition to mitochondrial functional assays, transcriptomic, metabolomic and inflammatory inflammatory mediator analyses. Compared with CSP, MP animals had: (1) significantly higher survival (100%vs. 33%; p<0.05); (2) superior graft function (p<0.05);(3) eight times higher hepatic O2 delivery than O2 consumption (0.78 mL O2/g/h vs. 0.096 mL O2/g/h) during MP; and (4) significantly greater bile production (MP=378.5 ± 179.7; CS=151.6 ± 116.85). MP downregulated interferon (IFN)-α and IFN-γ in liver tissue. MP allografts cleared lactate, produced urea, sustained gluconeogenesis and produced hydrophilic bile after reperfusion. Enhanced oxygenation under subnormothermic conditions triggers regenerative and cell protective responses resulting in improved allograft function. MP at 21 °C with the HBOC solution significantly improves liver preservation compared to CSP.

Evaluation of Calypte AWARE HIV-1/2 OMT antibody test as a screening test in an Indian setting.

PURPOSE: Integrated counselling and testing centres (ICTC) provide counselling and blood testing facilities for HIV diagnosis. Oral fluid tests provide an alternative for people who do not want blood to be drawn. Also, it avoids the risk of occupational exposure. The goal of this study was to evaluate the utility of Calypte AWARE HIV-1/2 OMT antibody test as a screening test in an Indian setting. MATERIALS AND METHODS: A cross-sectional study was carried out after ethics committee approval in 250 adult ICTC clients. Blood was collected and tested from these clients for HIV diagnosis as per routine policy and the results were considered as the gold standard. Also, after another written informed consent, oral fluid was collected from the clients and tested for the presence of HIV antibodies. Twenty five clients who had and 25 clients who had not completed their secondary school education (Group A and Group B, respectively) were also asked to perform and interpret the test on their own and their findings and experiences were noted. RESULT: The sensitivity, specificity, PPV and NPV of the oral fluid antibody test were 100%, 98.51%, 94.11% and 100%, respectively. Seventy six percent of clients preferred oral fluid testing. Group B found it difficult to perform the test as compared to Group A and this difference was statistically significant (P ≤ 0.05). CONCLUSION: Oral fluid testing can be used as a screening test for HIV diagnosis; however, confirmation of reactive results by blood-based tests is a must.

Study of HIV-1 subtypes in serodiscordant couples attending an integrated counselling and testing centre in Mumbai using heteroduplex mobility analysis and DNA sequencing.

AIMS: To determine the prevalent subtypes of HIV-1 in serodiscordant couples. SETTING: Integrated Counselling and Testing Centre (ICTC), Department of Microbiology. STUDY DESIGN: Prospective pilot study. PARTICIPANTS: Thirty HIV-1 serodiscordant couples. INCLUSION CRITERIA: a) Documentation of HIV-1 infection in one partner and seronegative status in the other, current history of continued unprotected sexual activity within the partnership, demonstration that they have been in a partnership for at least 1 year and are not currently on highly active antiretroviral therapy HAART; b) willingness of both partners to provide written informed consent including consent to continued couple counselling for 3 months. MATERIALS AND METHODS: HIV-1 subtyping was carried out by heteroduplex mobility analysis (HMA) by amplifying env region; and DNA sequencing by amplifying gag region. RESULTS: HIV-1 env gene was amplified successfully in 10/30 samples; gag gene, in 25/30 samples; and both env and gag gene were amplified successfully in 5/30 samples. HIV-1 subtype C was detected from 21 samples; subtype B, from 7; and subtype A, from 2. Sample from 1 positive partner was detected as subtype C by env HMA and subtype B by gag sequencing. CONCLUSION: HIV-1 subtype C was found to be the predominant subtype of HIV-1 in serodiscordant couples attending our ICTC, followed by HIV-1 subtype B and HIV-1 subtype A, respectively. DNA sequencing was found to be the most reliable method for determining the subtypes of HIV-1.

Detection of Japanese encephalitis virus by reverse transcription/polymerase chain reaction.

A simple and rapid reverse transcription/polymerase chain reaction (RT-PCR) assay for detection of Japanese encephalitis virus (JEV) envelope (E) gene sequences in various biological samples is described. The assay successfully amplified JEV E gene sequences from infected cell cultures, Aedes aegypti larvae, mosquitoes and mouse blood. The sensitivity of the assay was currently 1 ng of JEV RNA and could be increased up to 1 pg on the background of 1 microgram of cellular RNA by biotinylation of the PCR product, Southern blot analysis and streptavidin/alkaline phosphatase detection.

Phylogenetic analysis of the envelope gene of Japanese encephalitis virus.

Variation among Japanese encephalitis virus (JEV) strains has been documented in a number of studies by employing a variety of techniques like HI, NT, CF, RNA fingerprinting and sequencing of prM region. We report the complete envelope (E) gene sequence and the deduced amino acid sequence of four strains of JEV from the Indian subcontinent. These sequences were compared with published E gene sequences of 16 strains of JEV. Pairwise comparisons of the E gene nucleotide and deduced amino acid sequences of these strains indicated an overall sequence conservation. A majority of the differences in the four strains were located in domain A and domain C (Mandl et al., 1989). Phylogenetic analysis of the E gene sequences by a variety of tree building methods identified four clusters. Viral groupings did not correspond to geographic origin, isolation host or virulence. Evidence for positive selection operating on some strains belonging to different clusters was obtained.

Health care purchasing alliances--a Michigan multiemployer trust funds' approach.

A group of multiemployer trust funds in Michigan has formed a coalition that eventually will offer its members an integrated managed health care program. As the first step toward this goal, the coalition recently implemented a managed pharmaceutical program.

Charge anisotropy across biological membranes: evidence and implications.

Membrane proteins exhibit charge anisotropy across the bilayer with the vector positive inwards. The proton pumps, primary or secondary, which have been examined as a subset of these membrane proteins, also reveal charge anisotropy based on their sequence data. The direction of the anisotropy appears to satisfy the observed directional gradient of protons mediated by these proteins. A correct description of transport requires attention to local as well as field effects of the charge anisotropy of membrane proteins.

Pancreatic fibrosis-calcification with diabetes in South Maharashtra, India.

Out of 325 patients with diabetes mellitus 24 (7.4%) suffered from pancreatic fibrosis-calcification. Of these 21 were below the age of 30 years. Sorghum (Jowar) was the staple diet for all. Most patients belonged to the lowest socio-economic strata.