Correlation between MMP-9 and extracellular cytokine HMGB1 in prediction of human ischemic stroke outcome

AbstractIschemic stroke (IS) outcome predictors include clinical features, biochemical parameters and some risk factors. The relations between two main players in the ischemic brain, MMPs and HMGB1, were estimated in the plasma of ischemic stroke patients stratified according to the Glasgow Outcome Scale and the Oxfordshire Community Stroke Project classification. IS patients exhibited higher plasma concentration of MMP-9 and the inflammatory cytokine HMGB1 compared with healthy controls. A full-blown correlation between MMP-9 activation and increased plasma MMP-9 concentration was observed in case of IS patients. A similar activity of MMP-2 and MMP-12 was characteristic of healthy volunteers and IS patients. In patients with ischemic stroke increased plasma levels of MMP-9 and HMGB1 are associated with a poor functional outcome and are significantly correlated with each other (P=0.0054). We suggest that diagnostic benefits will be obtained if plasma HMGB1 levels are measured for IS patients in addition to MMP-9

A novel cassette method for probe evaluation in the designed biochips

A critical step in biochip design is the selection of probes with identical hybridisation characteristics. In this article we describe a novel method for evaluating DNA hybridisation probes, allowing the fine-tuning of biochips, that uses cassettes with multiple probes. Each cassette contains probes in equimolar proportions so that their hybridisation performance can be assessed in a single reaction. The model used to demonstrate this method was a series of probes developed to detect TORCH pathogens. DNA probes were designed for Toxoplasma gondii, Chlamidia trachomatis, Rubella, Cytomegalovirus, and Herpes virus and these were used to construct the DNA cassettes. Five cassettes were constructed to detect TORCH pathogens using a variety of genes coding for membrane proteins, viral matrix protein, an early expressed viral protein, viral DNA polymerase and the repetitive gene B1 of Toxoplasma gondii. All of these probes, except that for the B1 gene, exhibited similar profiles under the same hybridisation conditions. The failure of the B1 gene probe to hybridise was not due to a position effect, and this indicated that the probe was unsuitable for inclusion in the biochip. The redesigned probe for the B1 gene exhibited identical hybridisation properties to the other probes, suitable for inclusion in a biochip

Bioplastics against Microplastics: Screening of Environmental Bacteria for Bioplastics Production

Polyhydroxyalkanoates (PHAs) are biopolymers produced by numerous bacteria and can be used in the production of bioplastics. PHAs are synthesized by microorganisms by fermentation of carbon sources. Due to the different monomer structures of PHAs, there are many kinds of PHAs, and their corresponding material properties are also very different. Thus, the search for bacteria producing the PHAs is of great interest. In this study, the bacteria isolated from the environment were analyzed for the presence of PHA. PHA production was tested with staining methods Sudan Black B, Nile Blue, and Nile Red. The presence of a PHA synthase gene (phaC) was confirmed by PCR amplification. PHAs were extracted from the strains and characterized by the FTIR spectroscopy method. A biochip for a fast screening of environmental samples for the presence of PHA-producing bacteria was designed. The biochip contained 11 probes for coding class 1, 2, and 3 PHA synthase genes

Estimation of the Cellular Antioxidant Response to Chromium Action Using ESR Method

In the present study, the antioxidant capacity of chromium-treated L-41 (human epithelial-like cells) was investigated by the ESR spin-trapping technique. The crude cell extracts of the cells grown in the presence of 2 µM (nontoxic) and 20 µM (toxic) chromium (VI) concentrations were tested in the model Fenton system with and without catalase-inhibitor sodium azide. The presented approach using the ESR technique along with inhibitors lets us discern cell extract defense capacity connected with the enzymatic activity in viable cells and the catabolic activity in dying cells

Remedial Approaches against Arsenic Pollution

The study is devoted to a very urgent and acute problem for Georgia – remediation/restoration of the arsenic (As) mining and storage sites. The approach of a given work is based on using capabilities of nature itself, which has a great adaptive potential to chemical environmental pollution. The aim of the study is to identify the bacterial strains from the endemic soil microbiota, characteristic to a specific localization of arsenic contaminated sites and able to resist to the toxicant. To determine the level of arsenic contamination, soil samples have been analyzed using Inductively Coupled Plasma - Optical Emission Spectrometry method. The distribution of arsenic in soil samples splits them into categories according to the degree of contamination, ranging from 50 ppm to 13000 ppm. The local bacteria community has been studied using conventional cultivation method along with modern method of bioindication – a biochip. The low density biochip contains the relevant probes for the identification of the bacterial consortium in soil microbiota. Chemical and microbiological analysis was based on the standards and methodologies developed by International Standards Organizations – ISO and Environmental Protection Agency – EPA. It is prospected that bioremediation can become essential part of remediation against arsenic pollution in the context of circular economy

A Calorimetric Characterization of Cr(VI)-Reducing Arthrobacter oxydans

This is the first of a series of calorimetric studies designed to characterize and understand survival mechanisms of metal-reducing bacteria isolated from metal-polluted environments. In this paper we introduce a new concept of thermal spectrum of the endothermic melting of complex biological systems (e.g., proteins, nucleic acids, ribosomes, membrane structures) in intact cells. All thermal spectra measured are thermograms that describe the temperature dependence of heat capacity change of the complex systems of biologically active substances in bacterial cells. This new concept of thermal spectrum was applied to investigate spectral features from intact cells of Cr(VI)-reducer Arthrobacter oxydans at different points of their growth conditions and stages. Over the temperature range of 40–105°C, we observed that spectral changes are particularly significant in the 40–90°C interval. This may correspond to the orderly changes in subcellular structural elements: proteins, ribosomes and RNA, membranes, and various structural elements of the cell wall during different points of the growth cycle and growth conditions. Spectral changes in the 90–105°C region are less pronounced, implicating that the structural composition of DNA-Protein (DNP) complexes may change little

CK2 Phosphorylation of Schistosoma mansoni HMGB1 Protein Regulates Its Cellular Traffic and Secretion but Not Its DNA Transactions

parasite resides in mesenteric veins where fecundated female worms lay hundred of eggs daily. Some of the egg antigens are trapped in the liver and induce a vigorous granulomatous response. High Mobility Group Box 1 (HMGB1), a nuclear factor, can also be secreted and act as a cytokine. Schistosome HMGB1 (SmHMGB1) is secreted by the eggs and stimulate the production of key cytokines involved in the pathology of schistosomiasis. Thus, understanding the mechanism of SmHMGB1 release becomes mandatory. Here, we addressed the question of how the nuclear SmHMGB1 can reach the extracellular space. eggs of infected animals and that SmHMGB1 that were localized in the periovular schistosomotic granuloma were phosphorylated.We showed that secretion of SmHMGB1 is regulated by phosphorylation. Moreover, our results suggest that egg-secreted SmHMGB1 may represent a new egg antigen. Therefore, the identification of drugs that specifically target phosphorylation of SmHMGB1 might block its secretion and interfere with the pathogenesis of schistosomiasis

Экспрессия гена FABP4 в подкожной и висцеральной жировой ткани у пациентов с ожирением и сахарным диабетом II типа

Introduction. Obesity is associated with a high risk of developing concomitant diseases such as: metabolic syndrome, type 2 diabetes mellitus (DM2), cardiovascular pathology. FABP4 (fatty acid binding protein) is the specific lipid chaperone and an important protein for the function of adipose tissue and is one of the adipocytokines secreted by adipose tissue.The objective of the study was to investigate the FABP4 gene expression in subcutaneous and visceral adipose tissue (SAT and VAT) in patients with obesity and DM2.Methods and materials. SAT and VAT samples were obtained during bariatric surgery (N=43, BMI>35): obese with DM2 (N=21), obese without DM2 (N=22). Samples for the control group without obesity (N=15, BMI<30) were obtained during planned operations on the abdominal cavity. FABP4 mRNA level was estimated by real-time PCR.Results. It has been demonstrated that the mRNA level of the FABP4 gene in SAT and VAT is reduced in obesity, regardless of the manifestation of DM2 (p<0.01). A negative correlation was observed between the mRNA level of the FABP4 gene in adipose tissue and the BMI index (for SAT: r=—0.327, p = 0.016; for VAT: r=—0.304, p = 0.024).Conclusion. The expression level of FABP4 gene in AT can act as a marker of AT dysfunction in obesity.Введение. Ожирение ассоциировано с высоким риском развития таких заболеваний, как метаболический синдром, сахарный диабет II типа (СД2), сердечно-сосудистая патология. Специфический липидный шаперон FABP4 — белок, связывающий жирные кислоты, —является важным для функционирования жировой ткани белком, который при этом входит в число секретируемых жировой тканью адипоцитокинов.Целью исследования явилось изучение экспрессии гена FABP4 в подкожной и висцеральной жировой ткани (ПЖТ и ВЖТ) у пациентов с ожирением и СД2.Методы и материалы. Образцы ПЖТ и ВЖТ были получены при проведении бариатрических операций (N=43, ИМТ>35): 21 пациент с СД2, 22 — без СД2; а также у лиц без ожирения при плановых операциях на брюшной полости (контрольная группа, N=15, ИМТ<30). Уровень мРНК гена FABP4 оценивали методом полимеразной цепной реакции в режиме реального времени.Результаты. Было продемонстрировано, что уровень мРНК гена FABP4 в ПЖТ и ВЖТ снижен при ожирении независимо от манифестации СД2 (p<0,01). Наблюдалась отрицательная корреляция между уровнем мРНК гена FABP4 в жировой ткани и показателем ИМТ (для ПЖТ: r=—0,327, p = 0,016; для ВЖТ: r=—0,304, p = 0,024).Заключение. Уровень экспрессии гена FABP4 в ЖТ может выступать как маркер дисфункции ЖТ при ожирении