Exploring risk factors and transmission dynamics of Hepatitis B infection among Indian families: Implications and perspective.

INTRODUCTION: Hepatitis B virus (HBV) is global health problem. Family members of HBV infected people are considered as high-risk groups due to frequent household transmission of HBV among contacts of HBsAg carriers. The present study aimed to investigate the intrafamilial transmission of HBV among family members of HBV-infected persons and to identify the risk factors for viral transmission in these setting. METHODS: 361 index cases and their 1083 family contacts were tested for markers of Hepatitis B, viz. HBsAg and HBcAb using commercial ELISA. The demographic details and risk factors for acquiring HBV infection among the family members were recorded using a structured questionnaire. RESULTS: The median (IQR) age of index cases and family members was 37 (27 - 48) and 26 (14 - 38) years, respectively. Among the screened family members, 9.23% (n = 100) members were positive for HBsAg and 32.75% (n = 355) were positive for HBcAb. At least one member of the family was affected in 229/361 (63.43%) index cases. Significantly lower percent of household contacts (9.23%, n = 100)were vaccinated against HBV.HBV transmission risk was significantly higher in families with more than four members(p < 0.0001). Multinomial logistics regression analysis for familial risk factors for transmission of HBV such asclose contact with carrier (aOR overt: 1.172, aOR occult: 1.173), sharing of bed/bedding (aOR overt: 1.258, aOR occult:1.264), personal hygiene items (aOR overt:1.260, aOR occult: 1.451), and eating in common utensils (aOR overt: 2.182, aOR occult: 1.307)were significantly associated with the transmission of HBV (p < 0.05). DISCUSSION: Close contact with carrier, sharing of bed/bedding or personal hygiene items and eating in common utensils were significantly associated with the transmission of HBV. Increasing awareness about Hepatitis B infection and vaccination of family members in close contact with carrier is essential to prevent Hepatitis B transmission.

Asialoglycoprotein receptor targeted delivery of doxorubicin nanoparticles for hepatocellular carcinoma.

We report asialoglycoprotein receptor (ASGPR)-targeted doxorubicin hydrochloride (Dox) nanoparticles (NPs) for hepatocellular carcinoma (HCC). Polyethylene sebacate (PES)-Gantrez® AN 119 Dox NPs of average size 220 nm with PDI < 0.62 and ∼20% Dox loading were prepared by modified nanoprecipitation. ASGPR ligands, pullulan (Pul), arabinogalactan (AGn), and the combination (Pul-AGn), were anchored by adsorption. Ligand anchoring enabled high liver uptake with a remarkable hepatocyte:nonparenchymal cell ratio of 85:15. Furthermore, Pul-AGn NPs exhibited an additive effect implying incredibly high hepatocyte accumulation. Galactose-mediated competitive inhibition confirmed ASGPR-mediated uptake of ligand-anchored NPs in HepG2 cell lines. Subacute toxicity in rats confirmed the safety of the NP groups. However, histopathological evaluation suggested mild renal toxicity of AGn. Pul NPs revealed sustained reduction in tumor volume in PLC/PRF/5 liver tumor-bearing Nod/Scid mice up to 46 days. Extensive tumor necrosis, reduced collagen content, reduction in the HCC biomarker serum α-fetoprotein (p < 0.05), a mitotic index of 1.135 (day 46), and tumor treated/tumor control (T/C) values of <0.42 signified superior efficacy of Pul NPs. Furthermore, weight gain in the NP groups, and no histopathological alterations indicated that they were well tolerated by the mice. The high efficacy coupled with greater safety portrayed Pul Dox NPs as a promising nanocarrier for improved therapy of HCC.

Rapid flow cytometry based cytotoxicity assay for evaluation of NK cell function.

Assessment of natural killer cells (NK-cell) cytotoxicity is used not only in research settings but is also important in diagnosis of various diseases. NK-cell cytotoxicity assays are based on measurement of target cells killed by cytotoxic cells analyzed either by chromium (51Cr) release assay or flow cytometry. Both these methods use peripheral blood mononuclear cells (PBMC) or pure NK-cell population and hence require large volume of blood sample which is difficult to obtain in pediatric patients and patients with cytopenia. Hence, a flow cytometric assay was designed to determine NK cell activity using whole blood, eliminating the need for isolation of PBMCs or pure NK cells. This assay is based on a dual fluorescent staining of target cells (K562 cell line). The DIOC18 dye labeled K562 cells are incubated with whole blood and then counterstained with 7-AAD enabling the measurement of dead target cell and then percent cytotoxicity is calculated. This study compared the NK cell cytotoxicity using PBMC and whole blood in clinically relevant samples. There was no significant difference between two assays in the measurement of lytic activity or in-reproducibility in the repeated samplings of healthy individuals. The whole blood assay required less volume of blood and also less processing time as compared to PBMC assay. It was also validated by testing patients diagnosed with familial hemophagocytic lymphohistiocytosis expected to have low NK-cell activity. This assay is rapid, sensitive and reproducible and requires significantly less volume of blood which is important for clinical evaluation of NK-cell function.

Production of murine monoclonal antibody to fetal hemoglobin.

Fetal hemoglobin (Hb F) is a major hemoglobin (Hb) component at birth. Hb F levels are markedly elevated in a number of inherited and acquired disorders. Measurement of Hb F levels is usually carried out by alkali denaturation which is not very accurate for low and high values. An accurate estimation of Hb F, and also of F cells, is desired in many hematological disorders like sickle cell disease, in monitoring the efficacy of hydroxyurea (HU) therapy, to assess feto-maternal hemorrhage (FMH) during pregnancy and in the postpartum period. We have raised a murine monoclonal antibody to human Hb F, that accurately measures the number of F cells by flow cytometry. The antibody was found to be potent and specific for F cells.