Cellular Enzyme Activity Research Articles

Curcumin Protects Pheochromocytoma Cells Against Hydrogen Peroxide-Induced Oxidative Stress

Objective: Oxidative stress represents a characteristic of neurodegenerative diseases, which may be alleviated by the functional food component—curcumin. However, the underlying mechanism remains unclear. The current study aimed to address how curcumin protects variant differentiated pheochromocytoma (PC12) cells against hydrogen peroxide (H2O2)-induced cell stress, using low- and high-differentiated PC12 cells, and validated in old-aged mice. Methods: The MTT assay was first carried out to test the cell viability upon H2O2 and/or curcumin treatment and cellular lipid peroxidation was assessed as cellular damages by quantifying the amount of cellular malondialdehyde. Cellular oxidative stress was determined by measuring the cellular and mitochondrial ROS levels, and total cellular antioxidant capacities were determined as the ABTS free radical scavenging ability (ABTS) and Fe3+ reduction ability of cells. The activities and expression of major antioxidant enzymes, including glutathione peroxidase (GPx), superoxidase dismutase (SOD) and catalase (CAT), were also tested. Furthermore, oxidative phosphorylation of mitochondria was assessed by measuring the ATP level and the ADP/ATP ratio. Finally, the antioxidant activity of curcumin was determined in 6-month-old mice by measuring cellular antioxidant capacity and antioxidant enzyme activities. Results: H2O2 caused oxidative damage and decreased cell viability, which was reversed by curcumin with different dosed effects in variant differentiated PC12 cells. Moreover, curcumin's protective effect was attributed to an enhanced antioxidant enzyme system. In consistent, curcumin recovered H2O2-deteriorated oxidative phosphorylation. Further studies verified that curcumin significantly improved the antioxidant capacity of old-aged mice, which may have elevated oxidative stress. Conclusion: Taken together, curcumin improved the antioxidant capacity of variant differentiated PC12 cells and old-aged mice, which was closely associated with the modulation of mitochondrial function and antioxidant enzyme system. This study may shed light on the molecular mechanism underlying the alleviation of neurodegenerative diseases by natural polyphenols.

Synergistic Antibiofilm Action of Cinnamomum verum and Brazilian Green Propolis Hydroethanolic Extracts against Multidrug-Resistant Strains of Acinetobacter baumannii and Pseudomonas aeruginosa and Their Biocompatibility on Human Keratinocytes.

The accumulated dental biofilm can be a source of oral bacteria that are aspirated into the lower respiratory tract causing ventilator-associated pneumonia in hospitalized patients. The aim of this study was to evaluate the synergistic antibiofilm action of the produced and phytochemically characterized extracts of Cinnamomum verum and Brazilian green propolis (BGP) hydroethanolic extracts against multidrug-resistant clinical strains of Acinetobacter baumannii and Pseudomonas aeruginosa, in addition to their biocompatibility on human keratinocyte cell lines (HaCaT). For this, High-performance liquid chromatography analysis of the plant extracts was performed; then the minimum inhibitory and minimum bactericidal concentrations of the extracts were determined; and antibiofilm activity was evaluated with MTT assay to prevent biofilm formation and to reduce the mature biofilms. The cytotoxicity of the extracts was verified using the MTT colorimetric test, evaluating the cellular enzymatic activity. The data were analyzed with one-way ANOVA and Tukey's tests as well as Kruskal-Wallis and Dunn's tests, considering a significance level of 5%. It was possible to identify the cinnamic aldehyde in C. verum and p-coumaric, caffeic, and caffeoylquinic acids as well as flavonoids such as kaempferol and kaempferide and Artepillin-C in BGP. The combined extracts were effective in preventing biofilm formation and reducing the mature biofilms of A. baumannii and P. aeruginosa. Moreover, both extracts were biocompatible in different concentrations. Therefore, C. verum and BGP hydroethanolic extracts have bactericidal and antibiofilm action against multidrug resistant strains of A. baumannii and P. aeruginosa. In addition, the combined extracts were capable of expressively inhibiting the formation of A. baumannii and P. aeruginosa biofilms (prophylactic effect) acting similarly to 0.12% chlorhexidine gluconate.

Uji Aktivitas Antioksidan Formulasi Sediaan Krim Tabir Surya Ekstrak Etanol Biji Buah Durian (Durio Zibethiunus Murr) Dengan Menggunakan Metode DPPH

Durian fruit seeds are seeds that contain chemical compounds, namely flavonoid compounds. Flavonoid compounds are active compounds that are include in the type of antioxidant intermedietes that act as antioxidants. The antioxidant mechanism of flavonoids is to capture free radicals and reactive oxygen species (ROS) directly, prevent the regeneration of ROS and can indirectly increase the activity of antioxidants and cellular antioxidant enzymes. This study aims to determine durian seed extract as a natural sunscreen and determine the formula for a good cream preparation. The method used in the study of durian seed extract (Durio Zibethiunus Murr) is the DPPH (1,1-diphenyl-2-pikrilhidrazil) method. Then the metod of extraction is using maceration with the differences in each formula for sunscreen cream namely 0%, 5%, 10% and 15%. The evaluations used in the study were organoleptic tests, homogeneity tests, pH tests, adhesion tests and spreadability tests. The result of this study indicate that the formulation of sunscreen cearm extract of durian fruit seeds (Durio Zibethiunus Murr) can be used as a sunscreen cream. The best sunscreen cream formulation is the 15% formula. The content of the antioxidant compounds ir more and meets the ultra value.

Characterization of the Synergistic Antioxidant Activity of Epigallocatechin Gallate (EGCG) and Kaempferol.

Epigallocatechin gallate (EGCG) and kaempferol exhibit cellular antioxidant activity; however, their interactive effects in terms of antioxidant actions and underlying mechanisms remain unclear. In this study, their cytoprotective effects were examined against 2,2-azobis (2-amidinopropane) dihydrochloride solution (ABAP)-induced oxidative stress in HepG2 cells. The results showed that the median effective dose (EC50) of the EGCG and kaempferol (6:1.5, c/c) combination was 3.4 ± 0.1 μg/mL, with a combination index (CIavg) value of 0.54, which represented a significant synergistic effect. Further experiments proved that the combined pretreatment with EGCG and kaempferol exerted protective effects by suppressing reactive oxygen species (ROS) generation, upregulating cellular antioxidant enzyme activities (superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px)) in a dose-dependent manner. The mechanism of synergistic antioxidant effects of EGCG combined with kaempferol may be due to the up-regulation of higher antioxidant enzyme activities that improve the antioxidant capacities and balance the cell oxidative stress. The synergistic antioxidant effect of EGCG and kaempferol can provide a theoretical basis for the development of formulas of functional food ingredients.

Atovaquone exerts its anticancer effect by inhibiting Na+/K+-ATPase ion transport in canine cancer cells.

New anticancer drugs are being developed to avoid the toxicity and chemoresistance of the currently available drugs. The Food and Drug Administration-approved anti-malarial drug atovaquone is known to act as a selective oxidative phosphorylation inhibitor in the mitochondria by competing with CO Q10 (mitochondrial complex II and III). This study aimed to investigate the effect of atovaquone by examining the Na+/K+-ATPase (NKA) activity in various canine cell lines. Canine cell lines were treated with various concentrations (2.5, 5, 10, 15, and 20 μM) of atovaquone for 24, 48, and 72 h. Human cell lines were used as a control to validate the canine cancer cell lines. The activities of the drugs against the cancer cell lines were measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromideassay. The cell metabolic activity was determined by measuring the activities of the nicotinamide adenine dinucleotide phosphate-dependent cellular oxidoreductase enzymes. The NKA activity was measured using the single-cell patch clamping assay. Atovaquone-induced apoptosis by elevating the concentration of reactive oxygen species (ROS) in the tumor cells, leading to cell death. Treatment of canine cancer cells with N-acetylcysteine (ROS inhibitor) reduced the activity of the drug. Furthermore, atovaquone inhibited more than 45% of the NKA ion current. This study demonstrated effects of atovaquone against canine cancer cell lines. The data may prove beneficial in repurposing the drug as a new anticancer agent in canine clinical trials, which might aid in fighting human cancer.

Functional Characterization on Wheat Protein Kinase Gene <i>TaSnRK2.1</i> in Mediating Plant Resistance to Drought Stress

The protein kinase type of sucrose non-fermenting-1 related protein kinase 2 (SnRK2) plays an important role in regulating plant growth and development and in mediating plant resistance to abiotic stresses given that it involves transduction of distinct internal and abiotic stress signaling. In this study, <i>TaSnRK2.1</i>, a gene of SnRK2 family in wheat, was investigated focusing on its molecular characterization, expression pattern upon drought stress, and the biological function in mediating plant drought tolerance. At nucleic acid level, <i>TaSnRK2.1</i> is highly homologous to its counterparts of SnRK2 kinase family genes in species of Polish wheat and barley, whose translated protein harbors a conserved catalytic domain of Ser/Thr kinase domain (aa 62-aa 318). Under a 48 h-regime drought treatment, the transcripts of <i>TaSnRK2.1</i> showed obvious response in roots and leaves, with the highest expression level reached at 24 h after treatment and maintaining a high level to 48 h under drought condition. Meanwhile, the transcripts of <i>TaSnRK2.1</i> in both roots and leaves induced by drought were recovered following the normal recovery treatment. Gene transgene analysis indicated that compared with wild type control (WT), the tobacco line with sense-overexpression of <i>TaSnRK2.1</i> (Sen 1) enlarged plant growth phenotypes, increased dry weights, promoted stomata closing, increased contents of cellular osmtic-regulatory substances including proline, soluble protein, and soluble sugar, enhanced photosynthetic capacity and elevated cellular protective enzyme activities under drought treatment. In contrast, the line with antisense-expression of <i>TaSnRK2.1</i> (Anti 1) reduced plant growth phenotypes, decreased biomass, slowed stomata closing, decreased the contents of cellular osmtic-regulatory substances, photosynthetic capacity and protective enzyme activities compared with WT plants treated by drought stress. Our investigation suggested that <i>TaSnRK2.1</i> is sensitive in response to drought signaling at transcriptional level that confers enhanced plant drought resistance by regulating physiological processes associated with stomata movement, cellular osmotic-regulatory substance metabolism, photosynthetsis, and protective enzyme activities.

Distinctive Labeling of Live Monocytes and Neutrophils with a Single Fluorescent Molecule

(1) Background: a small-molecule fluorescent chemosensor, CDr20, tracks the resident macrophages based on the UGT1A7C activity in the brain, raising the possibility that additional immune cells expressing the UGT1A7C can be labeled with CDr20. (2) Methods: we applied CDr20 to various types of blood cells derived from hematopoietic organs (spleen and bone marrow) as well as peripheral blood to test the degree and selectivity of labeling of CDr20 in these cell types; (3) Results: CDr20 fluorescently labels monocytes/macrophages and neutrophils as a result of glucuronidation reaction (CDr20-Gluc), which is mediated with UGT1A7C. The selectivity of CDr20 labeling highly correlates with the Ugt1a7c expression level in immune cells. Moreover, CDr20-Gluc is exported from cells by a mechanism of how glucuronides within cells are excreted into extracellular space. Interestingly, the exportation of CDr20-Gluc is mainly observed in monocytes, potentially due to the monocyte-specific expression of ABCC transporters and this resulted in large differences in the degree of fluorescence retention in neutrophils (CDr20bright), compared to monocytes (CDr20dim) upon one hour of CDr20 incubation; (4) Conclusions: CDr20 can differentially label monocytes and neutrophils due to the variance in two different cellular enzymatic activities of UGT1A7C and ABCC. By using this property, CDr20 can be used to distinguish specific cell types within blood.

Development of biomaterial-based oxygen transportation vehicles for circulation within blood

BackgroundSeveral blood replacements have been developed because of issues with blood collection and storage, increased expenditures, and a global lack of reserves. Studies on artificial blood primarily concentrate on the research and synthesis of molecules that transport oxygen. The primary objective of the current work was to create a supporting, oxygen-carrying biological material that can replace blood and be used to save lives when blood is not available. MethodsThe work started with the synthesis of superparamagnetic iron oxide nanoparticles (SPIONS), and two distinct procedures were applied to ensure binding to hemoglobin (Hb) molecules. The surface modifications of the particles were carried out gradually by 3-amino propyl trimethoxy silane (3APGASPIONS) and tartaric acid (TASPIONS). Analysis methods used to characterize the particles include Fourier transform infrared spectrophotometer (FT-IR), elemental analysis (EDX), scanning electron microscope (SEM), X-ray diffractometry (XRD), and high contrast transmission electron microscopy (C-TEM). The characterization studies of SPIONS which bind Hb molecules to the surface were repeated and their effects on endothelial cell culture were examined to determine their toxic effects in vitro. Cell proliferation, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) activity changes were assessed in the evaluation of the possible toxic effects of the cells against the treated particles in the prepared cell lines. Blood substitute solutions synthesized by two different methods were stored on the shelf for about 1 month and the results were compared with fresh Hb solutions. Results and ConclusionThe CV and UV analysis results obtained from artificial Hb substitutes revealed that the structural integrity of Hb molecules did not change, the oxygen transport capabilities of biomolecules were preserved, and this state remained stable for a long time. Furthermore, it was determined that nanocomplexes synthesized by the two methods caused different cellular proliferation and there was a more limited increase in cellular enzyme activities (SOD, CAT, GPx) of Hb-bound particle solutions when compared to pure SPIONS.

Evaluation of Some Safety Parameters of Dual Histamine H3 and Sigma-2 Receptor Ligands with Anti-Obesity Potential.

Many studies have shown the high efficacy of histamine H3 receptor ligands in preventing weight gain. In addition to evaluating the efficacy of future drug candidates, it is very important to assess their safety profile, which is established through numerous tests and preclinical studies. The purpose of the present study was to evaluate the safety of histamine H3/sigma-2 receptor ligands by assessing their effects on locomotor activity and motor coordination, as well as on the cardiac function, blood pressure, and plasma activity of certain cellular enzymes. The ligands tested at a dose of 10 mg/kg b.w. did not cause changes in locomotor activity (except for KSK-74) and did not affect motor coordination. Significant reductions in blood pressure were observed after the administration of compounds KSK-63, KSK-73, and KSK-74, which seems logically related to the increased effect of histamine. Although the results of in vitro studies suggest that the tested ligands can block the human ether-a-go-go-related gene (hERG) potassium channels, they did not affect cardiac parameters in vivo. It should be noted that repeated administration of the tested compounds prevented an increase in the activity of alanine aminotransferase (AlaT) and gamma-glutamyl transpeptidases (gGT) observed in the control animals fed a palatable diet. The obtained results show that the ligands selected for this research are not only effective in preventing weight gain but also demonstrate safety in relation to the evaluated parameters, allowing the compounds to proceed to the next stages of research.

A combination fermentation strategy for simultaneously increasing cellular NADP(H) level, biomass, and enzymatic activity of glufosinate dehydrogenase in Escherichia coli

Oxidoreductase is one of the most important biocatalysts for the synthesis of various chiral compounds. However, their whole-cell activity is frequently affected by an insufficient supply of expensive nicotinamide cofactors. This study aimed to overcome such shortcomings by developing a combination fermentation strategy for simultaneously increasing intracellular NADP(H) level, biomass, and glufosinate dehydrogenase activity in E. coli. The results showed that the feeding mode of NAD(H) synthesis precursor and lactose inducer had essential effects on the accumulation level of intracellular NADPH. Adding 40mg L-1 of L-aspartic acid to the medium increased the intracellular NADP(H) concentration by 36.3%. Under the pH-stat feeding mode and adding 0.4g L-1h-1 lactose, the NADP(H) concentration, biomass, and GluDH activity in the 5-L fermenter reached 445.7μmol L-1, 21.7 gDCWL-1, and 8569.3UL-1, respectively. As far as we know, this is the highest reported activity of GluDH in the fermentation broth. Finally, the 5000-L fermenter was successfully scaled up to use this fermentation approach. The combination fermentation strategy might serve as a useful approach for the high-activity fermentation of other NADPH-dependent oxidoreductases.

Abstract 6050: Analysis of diverse metabolic pathways using bioluminescent cell-based assays

Abstract Cellular energy metabolism is a complex system of many interconnected metabolic pathways that govern both cell health and function. Cancer cells are adept at quickly switching between metabolic pathways to adapt to nutrient limitations in the environment, enhancing their growth advantages while negatively impacting other cells in the tumor microenvironment such as immune cells. To facilitate metabolic characterization of cancer cells, we developed assays for detection of metabolites involved in key pathways, such as glycolysis and glutaminolysis, using a bioluminescent NADH detection technology coupled to specific dehydrogenases. We have now extended these assays to include metabolites involved in glycogen, lipid, ketone body and amino acid metabolism, as well as intermediates in the TCA cycle such as malate, isocitrate, and 2-hydroxyglutarate. A second core bioluminescent hydrogen peroxide detection system was also used to develop an assay for pyruvate. Here we describe the use of these assays to measure metabolite levels in cell lysates and cell culture media collected from cancer cell lines plated in 96- or 384-well plates and in 2D or 3D cultures. Changes in metabolite levels were monitored in response to altered media composition and drug treatments. Levels were also compared between different cell lines. In comparative studies of glioblastoma cell lines, 2-hydroxyglutarate production was higher in cells expressing mutant isocitrate dehydrogenase. By reconfiguring the NADH detection technology to measure cellular enzyme activity rather than metabolite levels, we were able to detect activity of dehydrogenases that participate in the glycolysis pathway, the TCA cycle, and pentose phosphate pathway. Assays for cellular malate dehydrogenase, isocitrate dehydrogenase and other dehydrogenase enzymes were sensitive, requiring ≤ 400 lysed cells per assay. The expanded collection of bioluminescent assays provides a way to quickly characterize the metabolic states and behaviors of cancer cells to help identify pathways that may present therapeutic targets. Citation Format: Donna Leippe, Natasha Karassina, Mike Valley, Kayla Sylvester, Jolanta Vidugiriene. Analysis of diverse metabolic pathways using bioluminescent cell-based assays [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6050.

Commentary: Clinical profile, demographic distribution, and outcomes of ocular siderosis: Electronic medical record-driven big data analytics from an eye care network in India.

Commentary: Clinical profile, demographic distribution, and outcomes of ocular siderosis: Electronic medical record-driven big data analytics from an eye care network in India.

Studying the redox status of paramecium caudatum cells under influence of molybdenum, zinc, copper oxide nanoparticles and synthetic antioxidants

Aims. To study the effect of CuO, ZnO and MoO3 nanoparticles on the vital functions of the single cell organism Paramecium caudatum; to analyse the activity of antioxidant cellular enzymes: SOD and CAT, as well as to evaluate the possibility of cells protection against oxidative stress by endogenous antioxidants. Methods. Paramecium caudatum cells were cultured under standard conditions for this test subject [3-4]. The toxicity of metal nanoparticles, PMСO (20-60 nm), PM ZnO (40-80 nm) and PM MoO3 (5-20 nm) (Laboratory of Physical Modelling of Two-Phase Flows, United Institute of High Temperatures, Russian Academy of Sciences) — was evaluated by determining the LC50 lethality index of the concentration at which 50% of Paramecium caudatum population was killed. Free-floating cells were then exposed for 48 h to sublethal concentrations of NPs metal oxides in moderately hard water with dissolved antioxidant. The efficacy of emoxipine (10 mg/ml), mexidol (50 mg/ml) ascorbic acid AA (25 mg/ml) and nicotinic acid NA (10 mg/ml) was investigated; for this purpose cell number, cell membrane condition, changes of intracellular organelle shape were recorded (Levenhuk C310 digital camera microscope, 3.1 Mpixel). Results. We found that the presence of metal oxides induced a significant decrease (p &lt; 0.05) in the number of viable Paramecium caudatum cells, in 24 h there were 58 % deaths from CuO, 43 % deaths from ZnO and 40 % deaths from MoO3, but in 48 h - 92 %, 88 % and 75 %, respectively. Copper oxide PM were the most toxic to cells, with an LC50 of 25 mg/l; for ZnO PM and MoO3 PM, the lethal concentration was equal and higher than 50 mg/l. negative pressure of metal-containing NPs led to osmotic disorganization, which was accompanied by an increase in vacuoles, abnormal bending and membrane rupture. Conclusions. Our results demonstrated that oxidative stress is one of the leading mechanisms of toxicity of PM oxides of transition metals such as copper, zinc and molybdenum. When the single-celled organism Paramecium caudatum was exposed to the studied NPs, destructive damages of membrane structures, changes in functions and morphology of organelles leading to cell death were observed. It should be noted that under such abiotic stress, the viability of Paramecium caudatum in general is the result of a complex interaction of the cells' own antioxidant system as well as the specific characteristics of NPs (size and solubility) and additional environmental factors, such as the presence of compounds with antioxidant activity in it.

The effects of Chinese herbal feed additives on physiological health and detoxification ability in the red claw crayfish, Cherax quadricarinatus, and evaluation of their safety

The side effects of Chinese herbal medicine on metabolism in aquatic animals have not been fully studied. This study focused on evaluating the effects of Astragalus polysaccharide, chlorogenic acid, and berberine on the physiological health and detoxification ability of Cherax quadricarinatus. A basic diet served as a control, and diets containing differing concentrations of Astragalus polysaccharide (1, 2, 4, 8 g/kg), chlorogenic acid (1, 2, 4, 8 g/kg), or berberine (0.1, 0.5, 2.5 g/kg) were prepared. One thousand and eight crayfish (28.83 ± 0.29 g) were randomly allocated to 12 groups of net cages for 14 days. Samples were taken 7 days after feeding with Chinese herb bioactive ingredients and 7 days after ceasing this feeding. The results showed that compared with the control group, the addition of Astragalus polysaccharide, chlorogenic acid, or berberine did not have a significant effect on the growth performance of each experimental group. Cellular immunity, humoral immunity and antioxidant enzyme activities were increased significantly, while the concentrations of three herbal bioactive ingredients in plasma and metabolic enzymes improved significantly. Moreover, the bioactive ingredients did not cause biological macromolecular damage or histological damage in the hepatopancreas and significantly reduced the content of malondialdehyde. However, high concentrations of berberine (2.5 g/kg) significantly increased alanine aminotransferase and aspartate aminotransferase activities in the hepatopancreas, which could have a potential risk of damage to the hepatopancreas. Our results indicated that Astragalus polysaccharide, chlorogenic acid, and 0.1 g/kg berberine can be used as safe feed additives to stimulate the immune response, antioxidant capacity and detoxification ability and protect the organism from damage under oxidative stress.

Transcriptome analysis of gall oak (Quercus infectoria): De novo assembly, functional annotation and metabolic pathways analysis

Gall oak (Quercus infectoria) is a native tree of Iran, whose gall extract is used to treat many diseases. The presence of abundant secondary metabolites with various bioactivities in this plant has made it medically important. Despite its medicinal value, due to the lack of genomic information, the biosynthetic pathways of these compounds in this species are still unknown. The current research was aimed at observing, characterizing, and investigating the biosynthetic pathways of these compounds in Q.infectoria. De novo transcriptome assembly was conducted using the RNA sequencing technique. A total of 89,335 unigenes were generated, of which 6928 unigenes showed differential expression in leaves compared to root tissue. Gene ontology examination of DEGs revealed GO-term enrichment was related to cellular processes and enzyme activity. KEGG enrichment analysis for DEGs showed that most unigenes were related to metabolic pathways and biosynthesis of secondary metabolites. Moreover, 39 families of transcription factors were identified, of which the C2H2, bZIP, bHLH, and ERF TFs had the highest frequency. In the absence of a reference genome, the overall study of transcriptome will provide a reference for future functional and comparative studies. Moreover, the data obtained from sequencing and de novo assembly can be a valuable scientific resource for Q.infectoria.

Comparative evaluation and prioritization of key influences on biodegradation of 2,2′,4,4′-tetrabrominated diphenyl ether by bacterial isolate B. xenovorans LB400

Polybrominated diphenyl ethers (PBDEs) are a class of persistent organic pollutants being widely distributed and harmful to human health and wildlife, and the development of sustainable rehabilitation strategies including microbial degradation is of great concern. Although the increasing number of bacteria, especially the broad-spectrum and potent aerobes have been isolated for the efficient removal of PBDEs, the external influences and the corresponding influential mechanism on biodegradation are not fully understood yet. Given the wide-spectrum biodegradability of aerobic bacterial isolate, B. xenovorans LB400 for PBDEs, the dual impacts of many pivotal factors including pH, temperature, presence of dissolved organic matter (DOM) and cadmium ion etc. were comprehensively revealed on biodegradation of 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47). Due to the structural resemblance and stimulation of specific enzyme activity in bacteria, the biphenyl as substrates showed the greater capacity than non-aromatic compounds in improving biodegradation. The individual adaptation to neutrality and cultivation at about 30 °C was beneficial for biodegradation since the bacterial cellular viability and enzyme activity was mostly preserved. Although it was possibly good for the induction of hormesis and favorable to enhance the permeability or bioavailability of pollutant, the exceeding increase of Cd2+ or DOM may not give the profitable increase of biodegradation yet for the detrimental effect. For biodegradation, the mechanistic relationship that took account of the integrative correlation with the influential factors was artfully developed using partial least square (PLS) regression technique. Relative to the most significant influence of culture time and initial concentration of BDE-47, the larger relevance of other factors primarily marked as pH and DOM was consecutively shown after the quantitative prioritization. This may not only help understand the influential mechanism but provide a prioritizing regulation strategy for biodegradation of BDE-47. The PLS-derived relationship was validated with the certain predictability in biodegradation, and could be used as an alternative to accelerate a priori evaluation of suitability or improve the feasibility of such bacteria in remediation of PBDEs in the environment.

Cellular immunity and enzymatic activity of lymphocytes in chronic rhinosinusitis and chronic pain syndrome

Cellular immunity and enzymatic activity of lymphocytes in chronic rhinosinusitis and chronic pain syndrome

Ветеринарно-санитарная характеристика мяса петухов кросса Смена 9 при клеточном и напольно-выгульном выращивании

The veterinary and sanitary indicators of meat of roosters of the cross Shift 9, grown with cellular (1st group) and outdoor-walking (2nd group) content, were studied. It was established that by the age of 42 days, the roosters of the 1st group reached a live weight of 2280 g, the 2nd group – 2914 g. At the same time, the average daily increase was 69,19 and 70,02 g. The diameter of muscle fibers for 42 days it reached 57,25 microns, the average daily thickening of the fibers was 1.19 microns. There were no significant differences in hematological parameters, but the number of erythrocytes in the rooster of the 1st group with a white stripe was higher by 0,16×1012/l; RDW – by 5,4 %; MCHC – less by 11,5 g/l. The serum level of enzymes in a rooster with a white stripe was higher, which indicates increased activity of cellular enzymes. According to the indicators of organoleptic evaluation, the meat of roosters of the 1st group exceeded the meat of the 2nd group by 0.4 points; estimates of meat broth – by 0,4 points, but was inferior in weight loss at cooking by 1,3 %. The meat of chickens of the 1st group had a pH higher by 0,18, the acid number of fat – by 0,13 mg/KOH, volatile fatty acids – by 0,29 mg/KOH. The water content in the pectoral muscles of the cell-reared chickens was less than in the outdoor-walking group by 0,42 %; protein – by 0,25; fat – by 0,21 %, but the total fat content in the carcasses of the 1st group of pets was greater by 0,11 %. According to microbiological indicators, the meat of carcasses met the requirements GOST 31468-2012. In 50 % of the studied roosters of both groups, an expansion of the diameter of the endomysium and perimysium was observed with the accumulation of fluid and the formation of different volumes of vacuoles in muscle fibers and infiltration by immunocompetent cells (leukocytes, phagocytes). One rooster of cellular content had fibrosis of muscle fibers and the proliferation of fatty and connective tissues in the endomysium and perimysium, i.e. the development of myopathy with a white stripe.

The Enrichment of Specific Hair Follicle-Associated Cell Populations in Plucked Hairs Offers an Opportunity to Study Gene Expression Underlying Hair Traits

Gene expression differences can assist in characterizing important underlying genetic mechanisms between different phenotypic traits. However, when population-dense tissues are studied, the signals from scarce populations are diluted. Therefore, appropriately choosing a sample collection method that enriches a particular type of effector cells might yield more specific results. To address this issue, we performed a polyA-selected RNA-seq experiment of domestic horse (Equus ferus caballus) plucked-hair samples and skin biopsies. Then, we layered the horse gene abundance results against cell type-specific marker genes generated from a scRNA-seq supported with spatial mapping of laboratory mouse (Mus musculus) skin to identify the captured populations. The hair-plucking and skin-biopsy sample-collection methods yielded comparable quality and quantity of RNA-seq results. Keratin-related genes, such as KRT84 and KRT75, were among the genes that showed higher abundance in plucked hairs, while genes involved in cellular processes and enzymatic activities, such as MGST1, had higher abundance in skin biopsies. We found an enrichment of hair-follicle keratinocytes in plucked hairs, but detected an enrichment of other populations, including epidermis keratinocytes, in skin biopsies. In mammalian models, biopsies are often the method of choice for a plethora of gene expression studies and to our knowledge, this is a novel study that compares the cell-type enrichment between the non-invasive hair-plucking and the invasive skin-biopsy sample-collection methods. Here, we show that the non-invasive and ethically uncontroversial plucked-hair method is recommended depending on the research question. In conclusion, our study will allow downstream -omics approaches to better understand integumentary conditions in both health and disease in horses as well as other mammals.

Antioxidant Potential of Diosmin and Diosmetin against Oxidative Stress in Endothelial Cells

Phlebotropic flavonoids, including diosmin and its aglycone diosmetin, are natural polyphenols widely used in the prevention and treatment of chronic venous insufficiency (CVI). As oxidative stress plays an important role in the development of pathophysiology of the cardiovascular system, the study aimed to investigate the protective effects of diosmin and diosmetin on hydrogen peroxide (H2O2)-induced oxidative stress in endothelial cells. The cells were pretreated with different concentrations of the flavonoid prior to the H2O2 exposure. The cell viability, the level of intracellular reactive oxygen species (ROS), the activity of cellular antioxidant enzymes-including superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase GPx-and the malondialdehyde (MDA) level were assessed. It was found that the H2O2-induced oxidative stress was ameliorated by diosmin/diosmetin in a concentration-dependent manner. The flavonoids restored the activity of cellular antioxidant enzymes and lowered the MDA level upregulated by the H2O2 exposure. These results indicate that diosmin and diosmetin may prevent oxidative stress in endothelial cells; therefore, they may protect against the development and progression of oxidative-stress-related disorders.