HSV Usurps Eukaryotic Initiation Factor 3 Subunit M for Viral Protein Translation: Novel Prevention Target

Prevention of genital herpes is a global health priority. B5, a recently identified ubiquitous human protein, was proposed as a candidate HSV entry receptor. The current studies explored its role in HSV infection. Viral plaque formation was reduced by ∼90% in human cells transfected with small interfering RNA targeting B5 or nectin-1, an established entry receptor. However, the mechanisms were distinct. Silencing of nectin-1 prevented intracellular delivery of viral capsids, nuclear transport of a viral tegument protein, and release of calcium stores required for entry. In contrast, B5 silencing had no effect on these markers of entry, but inhibited viral protein translation. Specifically, viral immediate early genes, ICP0 and ICP4, were transcribed, polyadenylated and transported from the nucleus to the cytoplasm, but the viral transcripts did not associate with ribosomes or polysomes in B5-silenced cells. In contrast, immediate early gene viral transcripts were detected in polysome fractions isolated from control cells. These findings are consistent with sequencing studies demonstrating that B5 is eukaryotic initiation factor 3 subunit m (eIF3m). Although B5 silencing altered the polysome profile of cells, silencing had little effect on cellular RNA or protein expression and was not cytotoxic, suggesting that this subunit is not essential for host cellular protein synthesis. Together these results demonstrate that B5 plays a major role in the initiation of HSV protein translation and could provide a novel target for strategies to prevent primary and recurrent herpetic disease

The use of apolipoprotein A-I as a component of serum-free nutrient medium for bone marrow cell culture

An important step in preparation of cells for cell therapy and tissue engineering is the cultivation of cells in vitro. The aim of this work is to study the effect of human apolipoprotein A-I (apoA-I) on the functional activity of cultured bone marrow cells and to show the possibility of using this protein instead of animal fetal serum. Material and methods. Bone marrow cells were cultured in 24-well plates in RPMI-1640 medium in a CO2 incubator at a temperature of 37 °C. The rate of incorporation of [14C]-leucine into the total cell protein and [3H]-thymidine into the DNA was used as an integral indicator of cell viability during cultivation. Results and discussion. It was found that the rate of DNA synthesis in bone marrow cells in the presence of apo A-I increased compared with the control group (without apo A-I) by 55 % after 8 hours, by 523 % after 24 hours and by 219 % after 48 hours. Under these conditions the rate of protein synthesis was also increased. The results indicate that the presence of apo A-I in serum-free culture medium preserves the functional activity of cultured bone marrow cells. Considering that the regulatory effect of apo A-I is achieved at a low protein concentration in the medium (15 μg/ml), isolation of apo A-I from the patient’s own blood serum will provide a practically safe nutrient medium for culturing autologous bone marrow cells with applications in personalized cell therapy and tissue engineering

Влияние Кортизола в комплексе с липопротеинами очень низкой плотности на развитие гепатомы на-1 и карциномы Эрлиха

Studies were conducted using ascetic HA-1 hepatomas and ascitic Ehrlich’s carcinoma cells. After passage of cells in mice, cell culture containing tumor-associated macrophages was studied. It was shown that the very low-density lipoprotein (VLDL)-cortisol complex significantly inhibited proliferative activity of tumor cells. The obtained constant of cortisol binding to VLDL indicated that the protein component of lipoproteins participated in the complex formation. Tumor-associated macrophages were suggested to play an important role in the formation of this complex. It was found that inhibition of tumor cell growth was related to apoptosis.Исследования проводились на клетках асцитной НА-1 гепатомы и асцитной карциномы Эрлиха. После пассажа клеток на мышах выделяли смешанную культуру клеток, содержащую опухоль-ассоциированные макрофаги. Показано, что в сокультуре комплекс ЛПОНП–кортизол значительно подавлял пролиферативную активность опухолевых клеток. Полученная константа связы- вания кортизола с ЛПОНП указывает на то, что в комплексообразовании принимает участие белковый компонент липопротеинов. Предполагается, что в образовании такого комплекса важную роль играют опухоль-ассоциированные макрофаги. Обнаружено, что механизм подавления роста опухолевых клеток связан с апоптозо

INFLUENCE OF CORTISOL AND VERY LOW-DENSITY LIPOPROTEINES ON THE DEVELOPMENT OF HA-1HEPATOMA AND EHRLICH’S CARCINOMA

Studies were conducted using ascetic HA-1 hepatomas and ascitic Ehrlich’s carcinoma cells. After passage of cells in mice, cell culture containing tumor-associated macrophages was studied. It was shown that the very low-density lipoprotein (VLDL)-cortisol complex significantly inhibited proliferative activity of tumor cells. The obtained constant of cortisol binding to VLDL indicated that the protein component of lipoproteins participated in the complex formation. Tumor-associated macrophages were suggested to play an important role in the formation of this complex. It was found that inhibition of tumor cell growth was related to apoptosis

Neural network modelling of the influence of channelopathies on reflex visual attention

This paper introduces a model of Emergent Visual Attention in presence of calcium channelopathy (EVAC). By modelling channelopathy, EVAC constitutes an effort towards identifying the possible causes of autism. The network structure embodies the dual pathways model of cortical processing of visual input, with reflex attention as an emergent property of neural interactions. EVAC extends existing work by introducing attention shift in a larger-scale network and applying a phenomenological model of channelopathy. In presence of a distractor, the channelopathic network’s rate of failure to shift attention is lower than the control network’s, but overall, the control network exhibits a lower classification error rate. The simulation results also show differences in task-relative reaction times between control and channelopathic networks. The attention shift timings inferred from the model are consistent with studies of attention shift in autistic children