Genotyping of hepatitis B virus isolates from Lahaul and Spiti district in Himachal Pradesh,India

Hepatitis B virus (HBV) has several genotypes. In the Indian population, genotypes A and D are the most frequent. HBV infection is hyper-endemic in the Lahaul and Spiti district in Himachal Pradesh; however, the virus genotype in this area is not known. We sequenced a 398-nucleotide segment of HBV genome that included parts of pre-S1/S2 and polymerase genes from 17 specimens from this district, and assigned a viral genotype to these. Of the 17 specimens studied, 13 (76% [95% confidence interval?=?50–92%]) showed the presence of genotype C HBV; the remaining four were genotype D (n?=?4; 24%) HBV. Prevalence of genotype C HBV was much higher in the district than in other parts of India. This may reflect the historical mixing of this population with that in China. Since genotype C has a higher risk of chronicity and mother-to-child transmission, prevention of HBV infection may need particular emphasis in this area.     

Improving diagnosis of primary cytomegalovirus infection in pregnant women using immunoblots

Human cytomegalovirus (CMV) is the most common infectious cause of mental disability in newborns of developed countries. Transmission of CMV from mother to baby is more frequent in maternal primary infection, although CMV reactivation causes more congenital infections overall. Current diagnostic tests for distinguishing primary and reactivation CMV have problems with interpretation and immunoblots may assist with diagnosis. Sera from 60 pregnant women were analyzed using conventional serology in parallel with a commercial immunoblot assay (using Recomblot, Mikrogen Diagnostik). Comparison of detection of CMV IgG, IgM, IgG avidity in maternal primary infection showed the immunoblot relative to conventional serology had sensitivity and specificity of 100% for IgG identification. The detection of IgM on immunoblot showed sensitivity of 75%, specificity of 62.5%, positive predictive value (PPV) of 81.8% and negative predictive value (NPV) of 52.6%. The immunoblot IgG avidity assay had sensitivity of 94.1%, with a PPV of 100% when identifying low avidity serum samples, and sensitivity of 100% with a PPV of 97.1% for high avidity serum samples. Overall agreement between conventional serology (IgM, IgG avidity) and immunoblot (IgM, IgG avidity) for detection of primary CMV infection was 65%. Although the immunoblot is effective in detecting IgG and determining IgG avidity, it showed no significant benefits in performance or utility as a first line diagnostic technique for IgM or primary CMV infection in pregnant women. J. Med. Virol. 85:315–319, 2013. © 2012 Wiley Periodicals, Inc.     

Evaluation of hydro-alcoholic extract of Eclipta alba for its anticancer potential: an in vitro study

Ethnopharmacological relevance

Eclipta alba is traditionally used as hepatoprotective agent. The study was designed to explore its antiproliferative activity on liver and other related cancer.

Aim of the study

The present study was designed to assess and establish the role of Eclipta alba as anti-cancer agent using HepG2, C6 glioma and A498 cell lines as model system.

Materials and methods

Antiproliferative and cytotoxic effects of the Eclipta alba hydroalcoholic extract (EAE) was determined using MTT assay. The expression level of NF-kB was analysed by western blotting and RT PCR. Gelatin zymography was done for gelatinase matrix metalloproteinases (MMP-2 and 9) analysis.

Results

EAE inhibited the cell proliferation in dose dependent manner in HepG2, A498 and C6 glioma cell lines with an IC50 of 22 ± 2.9, 25 ± 3.6 and 50 ± 8.7 μg/ml, respectively. The expression of MMP (2 and 9) was down-regulated with EAE treatment. DNA damage was observed following 72 h of extract treatment, leading to apoptosis. Additionally, the expression level of NF-kB was evaluated with western blotting and RT-PCR and was found to be down-regulated/inactivated.

Conclusions

The data establish the existence of anti-proliferative, DNA damaging and anti-metastasis properties in EAE which is yet unexplored and hold high therapeutic impact.     

Changes in viral protein function that accompany retroviral endogenization

Endogenous retroviruses (ERVs) are the remnants of ancient retroviral infections of germ cells and have been maintained in whole or part as heritable genomic elements. The last known endogenization events occurred several million years ago, and therefore stepwise analysis of retroviral endogenization has not been possible. A unique opportunity to study this process became available when a full-length ERV isolated from koalas (KoRV) was shown to have integrated into their germ line within the past 100 years. Even though KoRV shares 78% nucleotide identity with the exogenous and highly infectious gibbon ape leukemia virus (GALV), the infectivity of KoRV, like that of other ERVs, is substantially lower than that of GALV. Differences in the protein coding regions of KoRV that distinguish it from GALV were introduced into the GALV genome, and their functional consequences were assessed. We identified a KoRV gagpol L domain mutation as well as five residues present in the KoRV envelope (env) that, when substituted for the corresponding residues of GALV, resulted in vectors exhibiting substantially reduced titers similar to those observed with KoRV vectors. In addition, KoRV env protein lacks an intact CETTG motif that we have identified as invariant among highly infectious gammaretroviruses. Disruption of this motif in GALV results in vectors with reduced syncytia forming capabilities. Functional assessment of specific sequences that contribute to KoRV's attenuation from a highly infectious GALV-like progenitor virus has allowed the identification of specific modifications in the KoRV genome that correlate with its endogenization.     

A case of acute endothelial corneal transplant rejection following immunization with ChAdOx1 nCoV-19 coronavirus vaccine

A 28-year-old female who underwent an uneventful femtosecond laser enabled keratoplasty (FLEK) in her left eye presented with pain, redness, and blurring of vision in the operated eye two weeks after getting immunized with COVID-19 vector vaccine (ChAdOx1 nCoV19 Vaccine Recombinant COVISHIELD, AstraZeneca). Slit-lamp examination showed donor stromal edema with Descemet’s membrane folds and Khodadoust line (KP’s on endothelium) with anterior chamber cells and flare. The patient was diagnosed with acute corneal graft rejection and advised hourly topical steroids with cycloplegics and oral steroids. The patient responded to treatment and there was progressive reversal of graft rejection with the patient achieving best spectacle-corrected visual acuity (BSCVA) of 20/30 after five weeks of treatment. Our case highlights possible immune corneal graft rejection after COVID19 vaccination and the need to step up topical steroids before vaccination.     

Structural alterations of branched versus linear mixed-surfactant micellar systems with the addition of a complex perfume mixture and dipropylene glycol as cosolvent

Personal care products commonly contain perfume mixtures, consisting of numerous perfume raw materials (PRMs), and cosolvents. The lipophilicity and structure of an individual PRM is known to affect its localization within the surfactant self-assembly as well as the micellar geometry. However, because multiple PRMs are used in formulations, significant intermolecular interactions between the PRMs and between the PRMs and the surfactant tail may also influence the location of the PRMs and their effects on the self-assembly. Herein, two anionic/zwitterionic mixed-surfactant systems (sodium trideceth-2 sulfate (ST2S)/cocamidopropyl betaine (CAPB) and sodium laureth-3 sulfate/CAPB) were formulated with a cosolvent (dipropylene glycol (DPG)) and 12 PRMs of varying structures and lipophilicities. This 12 PRM accord is simpler than a fully formulated perfume but more complex than a single perfume molecule. The geometric variations in the self-assemblies were evaluated using small-angle neutron scattering, perfume head space concentrations were determined using gas chromatography-mass spectrometry, and perfume localization was identified using NMR spectroscopy. The addition of the perfume accord caused enlargement of the micelles in both surfactant systems, with a greater change observed for ST2S/CAPB formulations. Furthermore, the addition of DPG to ST2S/CAPB resulted in micelle shrinkage. The micelle geometries and PRM localization in the micelles were affected by the degree of branching in the surfactant tail.

Personal care products commonly contain perfume mixtures, consisting of numerous perfume raw materials (PRMs), and cosolvents. Depending on the molecular structures of the additives and surfactants, the geometry of the colloidal structures can be affected.