The Epidermal Growth Factor Receptor (EGFR) Promotes Uptake of Influenza A Viruses (IAV) into Host Cells

Influenza A viruses (IAV) bind to sialic-acids at cellular surfaces and enter cells by using endocytotic routes. There is evidence that this process does not occur constitutively but requires induction of specific cellular signals, including activation of PI3K that promotes virus internalization. This implies engagement of cellular signaling receptors during viral entry. Here, we present first indications for an interplay of IAV with receptor tyrosine kinases (RTKs). As representative RTK family-members the epidermal growth factor receptor (EGFR) and the c-Met receptor were studied. Modulation of expression or activity of both RTKs resulted in altered uptake of IAV, showing that these receptors transmit entry relevant signals upon virus binding. More detailed studies on EGFR function revealed that virus binding lead to clustering of lipid-rafts, suggesting that multivalent binding of IAV to cells induces a signaling platform leading to activation of EGFR and other RTKs that in turn facilitates IAV uptake

Clinical, hormonal and molecular-genetic characteristics of monogenic diabetes mellitus associated with the mutations in the <i>INS</i> gene

Background: Currently more than 50 mutations of the INS gene are known to affect the various stages of insulin biosynthesis in the beta cells of the pancreas. However only individual cases of diabetes mellitus (DM) associated with heterozygous mutations in the coding region of the INS gene were reported in Russian Federation. We report a group of patients with a clinical manifestation of DM caused by mutations in both coding and non-coding regions of the INS gene. The patients with a mutation in the intron of the INS gene are reported for the first time in Russian FederationMaterials and methods: 60 patients with an isolated course of neonatal DM (NDM), 52 patients with a manifestation of DM at the age of 7–12 months and the absence of the main autoimmune markers of type 1 DM, 650 patients with the MODY phenotype were included in the study. NGS technology was used for molecular genetic research. Author’s panel of primers (Custom DNA Panel) was used for multiplex PCR and sequencing using Ion Ampliseq™ technology. The author’s panel “­Diabetes Mellitus” included 28 genes (13 candidate genes of MODY and other genes associated with DM).Results: 13 heterozygous mutations were identified in 16 probands and 9 relatives. The majority of mutations were detected in patients with PNDM (18.75%) and in patients with an onset of DM at the age of 7–12 months (9.6%). Mutations in the INS gene were detected in 2 patients (0.3%) in the group with the MODY phenotype. Mutations in the INS gene were not detected in patients with transient NDM (TNDM). Analysis of clinical data in patients with PND and onset of diabetes at the age of 7–12 months did not show significant differences in the course of the disease. The clinical characteristics of the cases of MODY10 and diabetes caused by a mutation in the intron of the INS gene are reported in details.Conclusion: The role of INS gene mutations in NDM, MODY, and DM with an onset at the age of 7–12 months was analyzed in a large group of patients. The clinical characteristics of DM due to a mutation in the intron of the INS gene are reported for the first time in the Russian Federation

Патоморфоз заболеваний бронхолегочной системы у работающих в контакте с аэрозолями цветных металлов

Etiopathogenic features and diagnostic criteria of occupational diseases studying in workers engaged in colored metallurgy have been given in the paper. Polyvalent sensitization to metal allergens (nickel, chrome, beryllium, manganese) was found. A toxic effect of nickel on DNA was shown that could be used as a biomarker of exposure for biological monitoring in colored metallurgy workers. Biochemical investigations determined the main pathogenic mechanisms underlying pathomorphology of bronchopulmonary diseases caused by the exposure of colored metals, such as activation of lipid peroxidation, "proteolysis – antiproteolysis" imbalance, growing significance of infection. This is the first study demonstrating clinical and biochemical parallels between characteristics of development and course of respiratory pathology caused by the exposure of colored metals. Infectious, inflammatory, toxico-allergic, and destructive processes predominated in this pathology. Preventive and rehabilitation strategies have been developed.В статье представлены результаты изучения этиопатогенетических особенностей формирования профессиональных, производственно обусловленных заболеваний у работающих в цветной металлургии, определены критерии их диагностики. Выявлена поливалентная сенсибилизация к основным металлам-аллергенам (никель, хром, марганец, бериллий). Установлено токсическое влияние никеля на ДНК, что может служить биомаркером экспозиции и использоваться для биологического мониторинга у работающих в цветной металлургии.Результаты биохимических исследований определили основные патогенетические механизмы, лежащие в основе патоморфоза бронхолегочной патологии при воздействии аэрозолей цветных металлов: активация перекисного окисления липидов и дисбаланс в системе "протеолиз-антипротеолиз", возрастание роли инфекционного фактора. Впервые выявлены клинико-биохимические параллели между особенностями формирования и течением бронхолегочной патологии, сформировавшейся под воздействием аэрозоля цветных металлов с преобладанием инфекционно-воспалительного, токсико-аллергического, воспалительно-деструктивного компонентов, и биохимическим профилем организма. Разработаны методы профилактики и реабилитации

Direct reprogramming of human fibroblasts into dopaminergic neuron-like cells

Transplantation of exogenous dopaminergic neuron (DA neurons) is a promising approach for treating Parkinson's disease (PD). However, a major stumbling block has been the lack of a reliable source of donor DA neurons. Here we show that a combination of five transcriptional factors Mash1, Ngn2, Sox2, Nurr1, and Pitx3 can directly and effectively reprogram human fibroblasts into DA neuron-like cells. The reprogrammed cells stained positive for various markers for DA neurons. They also showed characteristic DA uptake and production properties. Moreover, they exhibited DA neuron-specific electrophysiological profiles. Finally, they provided symptomatic relief in a rat PD model. Therefore, our directly reprogrammed DA neuron-like cells are a promising source of cell-replacement therapy for PD

The Role of Brucella abortus I-206 Thermoextracts in L- and S-form in Shaping the Immune Response in White Mice

Background. Brucellosis is an acute infectious-allergic zoonotic disease in which sick farm animals are the main source of infection for humans. Immunization of animals and people on the territory of Russia is carried out by live vaccines, which, having a high immunogenicity, can cause clinical, hematological and immune changes. Therefore, the work on the design of new vaccines that provide high protection and do not possess reactogenicity, agglutinogenic and sensitizing activity is the current direction of the study. Methodology. The study was performed on 72 outbred white mice immunized with inactivated B. abortus 19 ВA vaccine and thermal extracts (TE) of B. abortus I-206 in L- and S-form. Morphofunctional changes in the regional lymph node, spleen, and thymus of experimental animals were examined on days 3, 7, 14, and 21 of the time of immunization. Results.The influence of single introductions thermoextracts B. abortus I-206 in L-and S-form in comparison with inactivated vaccine B. abortus 19 ВА. the removal or the morphohistologic indices of immune organs of experimental animals. Found that thermoextracts have the ability to activate immune adjustment within the structures of the spleen and lymph nodes, without causing side effects, including allergic reactions. Conclusions. Preparations obtained from B. abortus in the L- and S-forms have the ability to activate immunological rearrangement in immunocompetent organs without causing any side effects. The results indicate the promise of further research in this direction

Three Ubiquitin Conjugation Sites in the Amino Terminus of the Dopamine Transporter Mediate Protein Kinase C–dependent Endocytosis of the Transporter

Dopamine levels in the brain are controlled by the plasma membrane dopamine transporter (DAT). The amount of DAT at the cell surface is determined by the relative rates of its internalization and recycling. Activation of protein kinase C (PKC) leads to acceleration of DAT endocytosis. We have recently demonstrated that PKC activation also results in ubiquitylation of DAT. To directly address the role of DAT ubiquitylation, lysine residues in DAT were mutated. Mutations of each lysine individually did not affect ubiquitylation and endocytosis of DAT. By contrast, ubiquitylation of mutants carrying multiple lysine substitutions was reduced in cells treated with phorbol ester to the levels detected in nonstimulated cells. Altogether, mutagenesis data suggested that Lys19, Lys27, and Lys35 clustered in the DAT amino-terminus are the major ubiquitin-conjugation sites. The data are consistent with the model whereby at any given time only one of the lysines in DAT is conjugated with a short ubiquitin chain. Importantly, cell surface biotinylation, immunofluorescence and down-regulation experiments revealed that PKC-dependent internalization of multilysine mutants was essentially abolished. These data provide the first evidence that the ubiquitin moieties conjugated to DAT may serve as a molecular interface of the transporter interaction with the endocytic machinery