Isolation of Subtype 3c, 3e and 3f-Like Hepatitis E Virus Strains Stably Replicating to High Viral Loads in an Optimized Cell Culture System

The hepatitis E virus (HEV) is transmitted via the faecal–oral route in developing countries (genotypes 1 and 2) or through contaminated food and blood products worldwide (genotypes 3 and 4). In Europe, HEV subtypes 3c, 3e and 3f are predominant. HEV is the leading cause of acute hepatitis globally and immunocompromised patients are particularly at risk. Because of a lack of cell culture systems efficiently propagating wild-type viruses, research on HEV is mostly based on cell culture-adapted isolates carrying uncommon insertions in the hypervariable region (HVR). While optimizing the cell culture system using the cell culture-adapted HEV strain 47832c, we isolated three wild-type strains derived from clinical specimens representing the predominant spectrum of HEV in Europe. The novel isolates 14-16753 (3c), 14-22707 (3e) and 15-22016 (3f-like) replicate to high viral loads of 108, 109 and 106.5 HEV RNA copies/mL at 14 days post-inoculation, respectively. In addition, they could be kept as persistently infected cell cultures with constant high viral loads (~109 copies/mL) for more than a year. In contrast to the latest isolates 47832c, LBPR-0379 and Kernow-C1, the new isolates do not carry genome insertions in the HVR. Optimization of HEV cell culture identified amphotericin B, distinct salts and fetal calf serum (FCS) as important medium supplements. Overconfluent cell layers increased infectivity and virus production. PLC/PRF/5, HuH-7-Lunet BLR, A549 and HepG2/C3A supported replication with different efficiencies. The novel strains and optimized cell culture system may be useful for studies on the HEV life cycle, inactivation, specific drug and vaccine development

Characteristics of immune memory 10–15 years after primary hepatitis B vaccination

Background and aims: The definition of immune memory after hepatitis B vaccination is still under debate. Therefore, we analysed hepatitis B surface antigen (HBsAg)-specific memory in more detail by investigating the kinetics of humoral and cellular responses after hepatitis B booster vaccination. Methods: The anti-HBs kinetics of 23 individuals with anti-HBs titres below 10 IU/I, who had been vaccinated 10-15 years ago, was monitored at day 0, 3, 7,14 and 28 after booster vaccination. HBsAg-specific IFN gamma- and IL5-secreting cells in enriched CD4(+) fraction were measured at day 0, 7 and 28 post-booster by enzyme-linked immunospot assay (ELISpot). Results: 22 of 23 subjects showed similar anti-HBs kinetic curves, including 3 of 4 subjects who did not reach anti-HBs titres of 10 IU/I. The steep anti-HBs increase started between day 3 and 7 and peaked around day 14. A plateau or only minimal changes were visible between day 14 and 28. 17.4% of subjects showed pre-booster cellular responses, and this rate had increased to 47.8% and 56.5% after 7 and 28 days, respectively. The kinetic patterns of T cell responses differed considerably among subjects. A dominance of Th2 responses (IL5 secretion) over Th1 responses (IFN gamma secretion) could be observed. Conclusions: The presence of B cell memory could be shown by a typical anamnestic anti-HBs response curve after a booster dose in all but one individual. In contrast, T cell responses to booster vaccination, which occurred in approximately 50% of participants, were rather heterogeneous. (C) 2015 Elsevier Ltd. All rights reserved

Мотивационное управление персоналом на примере компании ООО "Мобител"

Цель дипломной работы: анализ системы мотивационного управления в организации и разработка направлений его совершенствования. В ходе работы проводились практические исследования персонала, выявлялся мотивационный тип, определялась степень информативности персонала организации о системе мотивации (мотивационная среда). Был проведен практический анализ соотношения типов мотивации с формами. В ходе исследования применялось несколько практических методик по формированию мотивационной программы в компании. Степень внедрения: ряд рекомендаций по совершенствованию мотивационного управления персоналом были внедрены в практику управления ООО "Мобител".The purpose of the thesis: analysis of the system of motivational management in the organization and development of directions for its improvement. In the course of the work, practical research of personnel was carried out, the motivational type was revealed, the degree of information of the personnel of the organization about the system of motivation (motivational environment) was determined. A practical analysis of the relationship between types of motivation and forms was carried out. In the course of the research, several practical methods were used to form a motivational program in the company. Degree of implementation: a number of recommendations for improving motivational management of personnel were introduced into the practice of the management of LLC "Mobitel"