The Role of TRP Channels in Sepsis and Colitis.

To date, several members of the transient receptor potential (TRP) channels which provide a wide array of roles have been found in the gastrointestinal tract (GI). The goal of earlier research was to comprehend the intricate signaling cascades that contribute to TRP channel activation as well as how these receptors' activity affects other systems. Moreover, there is a large volume of published studies describing the role of TRP channels in a number of pathological disorders, including inflammatory bowel disease (IBD) and sepsis. Nevertheless, the generalizability of these results is subject to certain limitations. For instance, the study of IBD relies on various animal models and experimental methods, which are unable to precisely imitate the multifactorial chronic disease. The diverse pathophysiological mechanisms and unique susceptibility of animals may account for the inconsistency of the experimental data collected. The main purpose of this study was to conduct a comprehensive review and analysis of existing studies on transient receptor potential (TRP) channels implicating specific models of colitis and sepsis, with particular emphasis on their involvement in pathological disorders such as IBD and sepsis. Furthermore, the text endeavors to evaluate the generalizability of experimental findings, taking into consideration the limitations posed by animal models and experimental methodologies. Finally, we also provide an updated schematic of the most important and possible molecular signaling pathways associated with TRP channels in IBD and sepsis.

Hypoxia and Intestinal Inflammation: Common Molecular Mechanisms and Signaling Pathways.

The gastrointestinal tract (GI) has a unique oxygenation profile. It should be noted that the state of hypoxia can be characteristic of both normal and pathological conditions. Hypoxia-inducible factors (HIF) play a key role in mediating the response to hypoxia, and they are tightly regulated by a group of enzymes called HIF prolyl hydroxylases (PHD). In this review, we discuss the involvement of inflammation hypoxia and signaling pathways in the pathogenesis of inflammatory bowel disease (IBD) and elaborate in detail on the role of HIF in multiple immune reactions during intestinal inflammation. We emphasize the critical influence of tissue microenvironment and highlight the existence of overlapping functions and immune responses mediated by the same molecular mechanisms. Finally, we also provide an update on the development of corresponding therapeutic approaches that would be useful for treatment or prophylaxis of inflammatory bowel disease.

Inflammatory Bowel Disease: Crosstalk between Histamine, Immunity, and Disease.

Inflammatory bowel disease (IBD) is increasingly recognized as a serious, worldwide public health concern. It is generally acknowledged that a variety of factors play a role in the pathogenesis of this group of chronic inflammatory diseases. The diversity of molecular actors involved in IBD does not allow us to fully assess the causal relationships existing in such interactions. Given the high immunomodulatory activity of histamine and the complex immune-mediated nature of inflammatory bowel disease, the role of histamine and its receptors in the gut may be significant. This paper has been prepared to provide a schematic of the most important and possible molecular signaling pathways related to histamine and its receptors and to assess their relevance for the development of therapeutic approaches.

Crystal structure and function of human nucleoplasmin (npm2): a histone chaperone in oocytes and embryos.

Human Npm2 is an ortholog of Xenopus nucleoplasmin (Np), a chaperone that binds histones. We have determined the crystal structure of a truncated Npm2-core at 1.9 Å resolution and show that the N-terminal domains of Npm2 and Np form similar pentamers. This allowed us to model an Npm2 decamer which may be formed by hydrogen bonds between quasi-conserved residues in the interface between two pentamers. Interestingly, the Npm2 pentamer lacks a prototypical A1-acidic tract in each of its subunits. This feature may be responsible for the inability of Npm2-core to bind histones. However, Npm2 with a large acidic tract in its C-terminal tail (Npm2-A2) is able to bind histones and form large complexes. Fluorescence resonance energy transfer experiments and biochemical analysis of loop mutations support the premise that nucleoplasmins form decamers when they bind H2A-H2B dimers and H3-H4 tetramers simultaneously. In the absence of histone tetramers, these chaperones bind H2A-H2B dimers with a single pentamer forming the central hub. When taken together, our data provide insights into the mechanism of histone binding by nucleoplasmins.

Polymorphism of toll-like receptor genes and autoimmune endocrine diseases.

In recent years, there has been a significant amount of interest and vigorous studies on mutations related to innate immunity receptor genes such as Toll-like receptors (TLR), which is driven by the identification of many associations between these mutations and development of various disorders leading, in particular, to autoimmune diseases. It has been proven that the occurrence of single nucleotide polymorphisms in DNA sequences encoding TLRs causes malfunction of some key signaling pathways, and, as a result, increases the risk of autoimmune diseases. The identification of these polymorphisms can lead to the understanding of the pathogenesis of autoimmune diseases, which subsequently will create effective methods for the prevention and treatment thereof. This article examines the current state of the art, in particular summarizes data on the role of polymorphisms in Toll-like receptor genes in a number of autoimmune endocrine diseases, including type 1 diabetes mellitus, Graves' disease and Hashimoto's autoimmune thyroiditis. The search for relevant scientific data was carried out by entering search queries based on keywords: TLR, SNP, autoimmunity, Graves' disease, Type 1 diabetes mellitus, Hashimoto's autoimmune thyroiditis. The search was conducted through PubMed, MEDLINE, Elsevier journals, Science Direct and Russian Index of Scientific Citation, as well as other highly cited publications on Genetics, Immunology, and Pathophysiology - related to the topic.

Cloning, purification and structure determination of the HIV integrase-binding domain of lens epithelium-derived growth factor.

Lens epithelium-derived growth factor (LEDGF)/p75 is the dominant binding partner of HIV-1 integrase in human cells. The crystal structure of the HIV integrase-binding domain (IBD) of LEDGF has been determined in the absence of ligand. IBD was overexpressed in Escherichia coli, purified and crystallized by sitting-drop vapour diffusion. X-ray diffraction data were collected at Diamond Light Source to a resolution of 2.05 Å. The crystals belonged to space group P21, with eight polypeptide chains in the asymmetric unit arranged as an unusual octamer composed of four domain-swapped IBD dimers. IBD exists as a mixture of monomers and dimers in concentrated solutions, but the dimers are unlikely to be biologically relevant.

Ribosomal protein L1 recognizes the same specific structural motif in its target sites on the autoregulatory mRNA and 23S rRNA.

The RNA-binding ability of ribosomal protein L1 is of profound interest since the protein has a dual function as a ribosomal protein binding rRNA and as a translational repressor binding its mRNA. Here, we report the crystal structure of ribosomal protein L1 in complex with a specific fragment of its mRNA and compare it with the structure of L1 in complex with a specific fragment of 23S rRNA determined earlier. In both complexes, a strongly conserved RNA structural motif is involved in L1 binding through a conserved network of RNA-protein H-bonds inaccessible to the solvent. These interactions should be responsible for specific recognition between the protein and RNA. A large number of additional non-conserved RNA-protein H-bonds stabilizes both complexes. The added contribution of these non-conserved H-bonds makes the ribosomal complex much more stable than the regulatory one.

Evaluation of potential West Nile virus vectors in Volgograd region, Russia, 2003 (Diptera: Culicidae): species composition, bloodmeal host utilization, and virus infection rates of mosquitoes.

Potential West Nile virus (family Flaviviridae, genus Flavivirus, WNV) vectors were assessed during 2003 at indoor and outdoor collection sites in urban Volgograd, Russia, and in three nearby towns and surrounding rural areas. In total, 9,182 female mosquitoes comprising 13 species in six genera were collected. Relative abundance and bloodmeal host utilization differed temporarily and spatially. During June and July in Volgograd, Aedes vexans (Meigen) (85.4%) and Culex p. pipiens L. (7.6%) were the two most abundant species collected indoors, whereas during August, Cx. p. pipiens was the dominant species, accounting for 87.9% of specimens collected. Two WNV-positive mosquito pools were detected in August: one pool was composed of Cx. p. pipiens and the other pool of Culex modestus Ficalbi. Anopheles messeae Falleroni, Aedes caspius (Pallas), Ae. vexans, Cx. modestus, and Cx. p. pipiens used both humans and birds as bloodmeal sources. In urban areas, 20.4% of the Cx. p. pipiens fed on humans, 58.1% fed on chickens, and six specimens were positive for both chicken and human blood. Culex p. pipiens collected from flooded basements were predominantly autogenous (91.7%), whereas adult females resting in buildings with dry basements were composed of 67.5% anautogenous and 32.5% autogenous specimens. Our data suggest that the primary WNV vectors in the Volgograd region were Cx. p. pipiens and Cx. modestus and that intense transmission of WNV to humans in urban areas during the epidemic of 1999 may have been facilitated by the abundance and concentration of anautogenous Cx. p. pipiens in multistory buildings. The role of autogenous Cx. p. pipiens in urban transmission remains unresolved.

The incidence of West Nile disease in Russia in relation to climatic and environmental factors.

Since 1999, human cases of West Nile fever/neuroinvasive disease (WND) have been reported annually in Russia. The highest incidence has been recorded in three provinces of southern European Russia (Volgograd, Astrakhan and Rostov Provinces), yet in 2010-2012 the distribution of human cases expanded northwards considerably. From year to year, the number of WND cases varied widely, with major WND outbreaks in 1999, 2007, 2010, and 2012. The present study was aimed at identifying the most important climatic and environmental factors potentially affecting WND incidence in the three above-mentioned provinces and at building simple prognostic models, using those factors, by the decision trees method. The effects of 96 variables, including mean monthly temperature, relative humidity, precipitation, Normalized Difference Vegetation Index, etc. were taken into account. The findings of this analysis show that an increase of human WND incidence, compared to the previous year, was mostly driven by higher temperatures in May and/or in June, as well as (to a lesser extent) by high August-September temperatures. Declining incidence was associated with cold winters (December and/or January, depending on the region and type of model). WND incidence also tended to decrease during year following major WND outbreaks. Combining this information, the future trend of WND may be, to some extent, predicted, in accordance with the climatic conditions observed before the summer peak of WND incidence.

Structural basis for LMO2-driven recruitment of the SCL:E47bHLH heterodimer to hematopoietic-specific transcriptional targets.

Cell fate is governed by combinatorial actions of transcriptional regulators assembling into multiprotein complexes. However, the molecular details of how these complexes form are poorly understood. One such complex, which contains the basic-helix-loop-helix heterodimer SCL:E47 and bridging proteins LMO2:LDB1, critically regulates hematopoiesis and induces T cell leukemia. Here, we report the crystal structure of (SCL:E47)bHLH:LMO2:LDB1LID bound to DNA, providing a molecular account of the network of interactions assembling this complex. This reveals an unexpected role for LMO2. Upon binding to SCL, LMO2 induces new hydrogen bonds in SCL:E47, thereby strengthening heterodimer formation. This imposes a rotation movement onto E47 that weakens the heterodimer:DNA interaction, shifting the main DNA-binding activity onto additional protein partners. Along with biochemical analyses, this illustrates, at an atomic level, how hematopoietic-specific SCL sequesters ubiquitous E47 and associated cofactors and supports SCL's reported DNA-binding-independent functions. Importantly, this work will drive the design of small molecules inhibiting leukemogenic processes.

Development of aptamer therapeutics.

The field of aptamer research is growing rapidly, with ∼230 papers using the word 'aptamer' published from January to June 2010. These reports cover many different applications ranging from tools to study protein function to potential diagnostic and therapeutic agents. In this review we will focus on the processes involved in isolating and developing aptamers as therapeutic compounds, using specific examples including the first aptamer therapeutic approved for use in humans (Pegaptanib or Macugen). We will also mention a few of the growing number of aptamer therapeutics in various stages of preclinical and clinical trial.

Epidemiology of West Nile infection in Volgograd, Russia, in relation to climate change and mosquito (Diptera: Culicidae) bionomics.

In 1999, there was the large outbreak of West Nile fever (WNF) in Southern Russia (>500 cases in the Volgograd Province). In 2000-2004, the WNF incidence rate decreased steadily to zero, but a new outbreak occurred in 2007 (64 cases). The analysis of historical climate data for Volgograd from 1900 to present showed that the years 1999 and 2007 were the hottest ones due to a very mild "winter" (Dec.-Mar.) and a hot "summer" (June-Sep.). There are up to 15 potential WNF vectors in Volgograd, but only Culex pipiens and Culex modestus are abundant in late summer, both in urban and rural settings. Only these species are naturally attracted to and feed on both humans and birds. The RNA of pathogenic WN virus genovariant was found by reverse transcriptase polymerase chain reaction only in Culex mosquitoes at the infection rate of about 0.04%. So these species may be considered as potential WNF "bridge vectors" between birds and humans as well as main vectors in sylvatic avain cycle. Their abundance in an epidemic season was higher in the years with a mild winter and a hot summer, so this phenomenon may serve as a connecting link between a climate and WNF epidemiology. These findings give some hints on the predisposing factors for WNF epidemic as well as the possibility to predict WNF outbreaks in the temperate climate zones.

Structure of the L1 protuberance in the ribosome.

The L1 protuberance of the 50S ribosomal subunit is implicated in the release/disposal of deacylated tRNA from the E site. The apparent mobility of this ribosomal region has thus far prevented an accurate determination of its three-dimensional structure within either the 50S subunit or the 70S ribosome. Here we report the crystal structure at 2.65 A resolution of ribosomal protein L1 from Sulfolobus acidocaldarius in complex with a specific 55-nucleotide fragment of 23S rRNA from Thermus thermophilus. This structure fills a major gap in current models of the 50S ribosomal subunit. The conformations of L1 and of the rRNA fragment differ dramatically from those within the crystallographic model of the T. thermophilus 70S ribosome. Incorporation of the L1-rRNA complex into the structural models of the T. thermophilus 70S ribosome and the Deinococcus radiodurans 50S subunit gives a reliable representation of most of the L1 protuberance within the ribosome.