Host Defense Proteins and Peptides with Lipopolysaccharide-Binding Activity from Marine Invertebrates and Their Therapeutic Potential in Gram-Negative Sepsis

Sepsis is a life-threatening complication of an infectious process that results from the excessive and uncontrolled activation of the host’s pro-inflammatory immune response to a pathogen. Lipopolysaccharide (LPS), also known as endotoxin, which is a major component of Gram-negative bacteria’s outer membrane, plays a key role in the development of Gram-negative sepsis and septic shock in humans. To date, no specific and effective drug against sepsis has been developed. This review summarizes data on LPS-binding proteins from marine invertebrates (ILBPs) that inhibit LPS toxic effects and are of interest as potential drugs for sepsis treatment. The structure, physicochemical properties, antimicrobial, and LPS-binding/neutralizing activity of these proteins and their synthetic analogs are considered in detail. Problems that arise during clinical trials of potential anti-endotoxic drugs are discussed

Studies on the Structure and Properties of Membrane Phospholipase A1 Inclusion Bodies Formed at Low Growth Temperatures Using GFP Fusion Strategy

The effect of cultivation temperatures (37, 26, and 18 °C) on the conformational quality of Yersinia pseudotuberculosis phospholipase A1 (PldA) in inclusion bodies (IBs) was studied using green fluorescent protein (GFP) as a folding reporter. GFP was fused to the C-terminus of PldA to form the PldA-GFP chimeric protein. It was found that the maximum level of fluorescence and expression of the chimeric protein is observed in cells grown at 18 °C, while at 37 °C no formation of fluorescently active forms of PldA-GFP occurs. The size, stability in denaturant solutions, and enzymatic and biological activity of PldA-GFP IBs expressed at 18 °C, as well as the secondary structure and arrangement of protein molecules inside the IBs, were studied. Solubilization of the chimeric protein from IBs in urea and SDS is accompanied by its denaturation. The obtained data show the structural heterogeneity of PldA-GFP IBs. It can be assumed that compactly packed, properly folded, proteolytic resistant, and structurally less organized, susceptible to proteolysis polypeptides can coexist in PldA-GFP IBs. The use of GFP as a fusion partner improves the conformational quality of PldA, but negatively affects its enzymatic activity. The PldA-GFP IBs are not toxic to eukaryotic cells and have the property to penetrate neuroblastoma cells. Data presented in the work show that the GFP-marker can be useful not only as target protein folding indicator, but also as a tool for studying the molecular organization of IBs, their morphology, and localization in E. coli, as well as for visualization of IBs interactions with eukaryotic cells

Complex estimation of paper printing properties for the offset printing

В даний час велику увагу приділяють оцінці якості продукції. У той же час існує проблема узгодженості даних виробника і споживача. Оцінка якості паперу для друку повинна містити не тільки аналіз його фізико-механічних і оптичних властивостей, а й комплекс друкарських властивостей з метою підвищення стабільності друку і виробництва паперу з поліпшеними друкарськими властивостями. У статті поставлено завдання не тільки аналізу і підвищення властивостей кінцевого продукту, але і прогнозування його властивостей. Для цього проведені випробування зі зміни композиційного складу паперу і його впливу на комплекс друкарських властивостей, визначених за технологією багатофарбового прободруку на основі аналізу зображення комплексної тестової шкали. У результаті отримана багатофакторна система із заданими параметрами композиції і виміряними даними показників якості друкованого зображення. Для зниження розмірності системи введений узагальнюючий критерій, який дозволив провести оптимізацію композиції з урахуванням факторів її варіювання і спрогнозувати підвищення друкарських властивостей в комплексі. У результаті проведення дослідно-промислових випробувань отримано папір з поліпшеними друкарськими властивостями.Now much attention is paid to assessing the products quality. At the same time there is a problem of consistency the data producers and consumers. Assessment of quality of the printing paper should include not only the analysis of its physical-mechanical properties and the optical, but also a complex of printing properties, in order to increase the stability of printing and producing a paper with an improved printing properties. The article concerns not only the analysis and enhancement of the properties of the final product, but also the prediction of its properties. For this purpose test conducted by changing the composition of paper and its effect on the printability complex defined by multicolor proof printing technology based on analysis of image of complex test scale. The result is a multi-factorial system with the specified parameters of the composition and quality characteristics of the printed image. To reduce the dimensionality of the system introduced by generalizing criterion that allowed for the optimization of the composition, taking into account the factors of its variation and forecast increase printing properties in the complex. As a result of pilot tests the paper with improved printing properties is got out.В настоящее время большое внимание уделяют оценке качества продукции. В то же время существует проблема согласованности данных производителя и потребителя. Оценка качества бумаги для печати должна включать не только анализ ее физико-механических и оптических свойств, но и комплекс печатных свойств с целью повышения стабильности печати и производства бумаги с улучшенными печатными свойствами. В статье поставлена задача не только анализа и повышения свойств конечного продукта, но и прогнозирования его свойств. Для этого проведены испытания по изменению композиционного состава бумаги и его влиянию на комплекс печатных свойств, определенных по технологии многокрасочного пробопечатания на основе анализа изображения комплексной тестовой шкалы. В результате получена многофакторная система с заданными параметрами композиции и измеренными данными показателей качества печатного изображения. Для снижения размерности системы введен обобщающий критерий, который позволил провести оптимизацию композиции с учетом факторов ее варьирования и спрогнозировать повышение печатных свойств в комплексе. В результате проведения опытно-промышленных испытаний получена бумага с улучшенными печатными свойствами

Porin from Marine Bacterium Marinomonas primoryensis KMM 3633T: Isolation, Physico-Chemical Properties, and Functional Activity

Marinomonas primoryensis KMM 3633T, extreme living marine bacterium was isolated from a sample of coastal sea ice in the Amursky Bay near Vladivostok, Russia. The goal of our investigation is to study outer membrane channels determining cell permeability. Porin from M. primoryensis KMM 3633T (MpOmp) has been isolated and characterized. Amino acid analysis and whole genome sequencing were the sources of amino acid data of porin, identified as Porin_4 according to the conservative domain searching. The amino acid composition of MpOmp distinguished by high content of acidic amino acids and low content of sulfur-containing amino acids, but there are no tryptophan residues in its molecule. The native MpOmp existed as a trimer. The reconstitution of MpOmp into black lipid membranes demonstrated its ability to form ion channels whose conductivity depends on the electrolyte concentration. The spatial structure of MpOmp had features typical for the classical gram-negative porins. However, the oligomeric structure of isolated MpOmp was distinguished by very low stability: heat-modified monomer was already observed at 30 °C. The data obtained suggest the stabilizing role of lipids in the natural membrane of marine bacteria in the formation of the oligomeric structure of porin