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

Development of Multiplex PCR Coupled DNA Chip Technology for Assessment of Endogenous and Exogenous Allergens in GM Soybean

Allergenicity assessment of transgenic plants and foods is important for food safety, labeling regulations, and health protection. The aim of this study was to develop an effective multi-allergen diagnostic approach for transgenic soybean assessment. For this purpose, multiplex polymerase chain reaction (PCR) coupled with DNA chip technology was employed. The study was focused on the herbicide-resistant Roundup Ready soya (RRS) using a set of certified reference materials consisting of 0, 0.1%, 0.5%, and 10% RRS. Technically, the procedure included design of PCR primers and probes; genomic DNA extraction; development of uniplex and multiplex PCR systems; DNA analysis by agarose gel electrophoresis; microarray development, hybridization, and scanning. The use of the asymmetric multiplex PCR method is shown to be very efficient for DNA hybridization with biochip probes. We demonstrate that newly developed fourplex PCR methods coupled with DNA-biochips enable simultaneous identification of three major endogenous allergens, namely, Gly m Bd 28K, Gly m Bd 30K, and lectin, as well as exogenous 5-enolppyruvyl shikimate-phosphate synthase (epsps) expressed in herbicide-resistant roundup ready GMOs. The approach developed in this study can be used for accurate, cheap, and fast testing of food allergens

Antioxidant Capacity of Cultured Mammalian Cells Estimated by ESR Method

In the present study, the antioxidant capacity against hydrogen peroxide (H2O2), one of the stress-inducing agents, was investigated in two distinct cell lines: L-41 (human epithelial-like cells) and HLF (human diploid lung fibroblasts), which differ in tissue origin, life span in culture, proliferate activity, and special enzyme system activity. The cell antioxidant capacity against H2O2 was estimated by the electron spin resonance (ESR) spin-trapping technique in the Fenton reaction system via Fe+2 ion action with H2O2 resulting in hydroxyl radical generation. The effects of catalase inhibitors, such as sodium azide and 3-amino-1,2,4-triazole, on the antioxidant capacity of cells were tested. Based on our observation, it can be concluded that the defensive capacity of cells against H2O2 depends on the ratio between catalase/GPx/SOD and H2O2, especially at high-stress situations, and the intracellular balance of these enzymes are more important than the influence of the single component

Characteristics of the probes used in this study.

<p>Characteristics of the probes used in this study.</p