Altered Enzyme Activity Research Articles

Effects of 2,4-di-tert-butylphenol at different concentrations on soil functionality and microbial community structure in the Lanzhou lily rhizosphere

The autotoxicity of root exudates is an important factor that affects soil enzymes and microbes in rhizosphere soil. Previous studies have confirmed that 2,4-di-tert-butylphenol (2,4-DTBP) is a major autotoxin of the root exudates that severely hampers the yield and quality of Lanzhou lily. However, little is currently known regarding the mechanism of 2,4-DTBP. Here, we hypothesized that 2,4-DTBP has a direct effect on soil functionality and the microbial community structure in Lanzhou lily rhizosphere soil, thereby affecting the growth of Lanzhou lily. To test our hypothesis, various experiments were conducted in this study. We treated Lanzhou lily plants with 2,4-DTBP at different concentrations and analyzed the effects of 2,4-DTBP on rhizosphere soil enzyme activity and microbial community structure and function. The diversity of soil microbes was analyzed using high-throughput sequencing. The results showed that 2,4-DTBP altered rhizosphere soil enzyme activity and microbial community structure and function. Moreover, microbial biomass was stimulated with the addition of 0.5 mM 2,4-DTBP and inhibited with the addition of 2.0 mM 2,4-DTBP. Thus, the 2.0 mM 2,4-DTBP treatment was not suitable for the growth of soil microorganisms. We also found that the 2.0 mM 2,4-DTBP treatment resulted in an increase in the levels of fungal pathogens, such as Fusarium, and reduced the size of beneficial bacterial populations in the rhizosphere soil. Moreover, the effects of 2,4-DTBP on soil functionality and the microbial community structure differed based on the concentrations of 2,4-DTBP and the specific indexes of determination. Thus, we concluded that 2,4-DTBP could inhibit soil functionality and disturb the microbial community structure in the rhizosphere soil. This finding may be important for understanding the relationship between autotoxicity and barriers and clarifying the long-term continuous cropping problems of Lanzhou lily.

Resistance to mesosulfuron-methyl in Beckmannia syzigachne may involve ROS burst and non-target-site resistance mechanisms

Herbicide resistance to chemical herbicide is a global issue that presents an ongoing threat to grain production. Though it has been frequently implicated that the production of detoxification enzymes increased in resistance development, the mechanisms for overexpression of these genes employed by herbicide-resistant weeds remain complicated. In this study, a mesosulfuron-methyl resistant Beckmannia syzigachne population (R) was found to be cross-resistant to another herbicide pyriminobac-methyl. No known target-site mutations were detected in the R population. In contrast, the decreased uptake and enhanced metabolic rates of mesosulfuron-methyl were detected in the R than the susceptible (S) population. Two candidate ATP-binding cassette (ABC) transporter genes (ABCB25 and ABCC14) that were constitutively up-regulated in the R population were identified by RNA-sequencing and validated by RT-qPCR. Alteration of antioxidant enzyme activities and gene expressions implied that mesosulfuron-methyl-induced antioxidant defenses provoked reactive oxygen species (ROS) burst. ROS scavenger assay showed that ROS induces ABCB25 and ABCC14 expression. This study reported for the first time that ABC transporters mediated non-target-site resistance contributes to mesosulfuron-methyl resistance in a B. syzigachne population, and implicated that ROS burst might be involved in the overexpression of ABC transporter genes in weeds.

Warming and Labile Substrate Addition Alter Enzyme Activities and Composition of Soil Organic Carbon

Warming can increase the efflux of carbon dioxide (CO2) from soils and can potentially feedback to climate change. In addition to warming, the input of labile carbon can enhance the microbial activity by stimulating the co-metabolism of recalcitrant soil organic matter (SOM). This is particularly true with SOM under invaded ecosystems where elevated CO2and warming may increase the biomass of invasive species resulting in higher addition of labile substrates. We hypothesized that the input of labile carbon would instigate a greater soil organic carbon (SOC) loss with warming compared to the ambient temperature. We investigated this by incubating soils collected from a native pine (Pinus taeda) forest to which labile carbon from the invasive species kudzu (Pueraria lobata) was added. We evaluated the microbial extracellular enzyme activity, molecular composition of SOC and the temperature sensitivity of soil CO2efflux under warming and labile carbon addition. After 14 months of soil incubation, the addition of labile C through kudzu extract increased the activity of β-1,4-glucosidase compared with the control. However, the activity of N-acetyl-β-D-glucosaminidase and fungal biomass (ergosterol) decreased with labile carbon addition. The activity of peroxidase increased with warming after 14 months of soil incubation. Although the carbon content of incubated soils did not vary with substrate and temperature treatments, the molecular composition of SOC indicated a general decrease in biopolymers such as cutin, suberin, long-chain fatty acids, and phytosterol with warming and an increasing trend of microbial-derived compounds with labile substrate addition. In soils that received an addition of labile C, the macro-aggregate stability was higher while the temperature sensitivity of soil C efflux was lower compared with the control. The increase in aggregate stability could enhance the physical protection of SOC from microbial decomposition potentially contributing to the observed pattern of temperature sensitivity. Our results suggest that warming could preferentially accelerate the decomposition of recalcitrant compounds while the addition of labile substrates could enhance microbial-derived compounds that are relatively resistant to further decomposition. Our study further emphasizes that global change factors such as plant invasion and climate change can differentially alter soil microbial activity and the composition of SOC.

PsEND1 Is a Key Player in Pea Pollen Development Through the Modulation of Redox Homeostasis.

Redox homeostasis has been linked to proper anther and pollen development. Accordingly, plant cells have developed several Reactive Oxygen Species (ROS)-scavenging mechanisms to maintain the redox balance. Hemopexins constitute one of these mechanisms preventing heme-associated oxidative stress in animals, fungi, and plants. Pisum sativum ENDOTHECIUM 1 (PsEND1) is a pea anther-specific gene that encodes a protein containing four hemopexin domains. We report the functional characterization of PsEND1 and the identification in its promoter region of cis-regulatory elements that are essential for the specific expression in anthers. PsEND1 promoter deletion analysis revealed that a putative CArG-like regulatory motif is necessary to confer promoter activity in developing anthers. Our data suggest that PsEND1 might be a hemopexin regulated by a MADS-box protein. PsEND1 gene silencing in pea, and its overexpression in heterologous systems, result in similar defects in the anthers consisting of precocious tapetum degradation and the impairment of pollen development. Such alterations were associated to the production of superoxide anion and altered activity of ROS-scavenging enzymes. Our findings demonstrate that PsEND1 is essential for pollen development by modulating ROS levels during the differentiation of the anther tissues surrounding the microsporocytes.

Effects of sulfuric, nitric, and mixed acid rain on the decomposition of fine root litter in Southern China

BackgroundChina has been increasingly subject to significant acid rain, which has negative impacts on forest ecosystems. Recently, the concentrations of NO3− in acid rain have increased in conjunction with the rapid rise of nitrogen deposition, which makes it difficult to precisely quantify the impacts of acid rain on forest ecosystems.MethodsFor this study, mesocosm experiments employed a random block design, comprised of ten treatments involving 120 discrete plots (0.6 m × 2.0 m). The decomposition of fine roots and dynamics of nutrient loss were evaluated under the stress of three acid rain analogues (e.g., sulfuric (SO42−/NO3− 5:1), nitric (1:5), and mixed (1:1)). Furthermore, the influences of soil properties (e.g., soil pH, soil total carbon, nitrogen, C/N ratio, available phosphorus, available potassium, and enzyme activity) on the decomposition of fine roots were analyzed.ResultsThe soil pH and decomposition rate of fine root litter decreased when exposed to simulated acid rain with lower pH levels and higher NO3− concentrations. The activities of soil enzymes were significantly reduced when subjected to acid rain with higher acidity. The activities of soil urease were more sensitive to the effects of the SO42−/NO3− (S/N) ratio of acid rain than other soil enzyme activities over four decomposition time periods. Furthermore, the acid rain pH significantly influenced the total carbon (TC) of fine roots during decomposition. However, the S/N ratio of acid rain had significant impacts on the total nitrogen (TN). In addition, the pH and S/N ratio of the acid rain had greater impacts on the metal elements (K, Ca, and Al) of fine roots than did TC, TN, and total phosphorus. Structural equation modeling results revealed that the acid rain pH had a stronger indirect impact (0.757) on the decomposition rate of fine roots (via altered soil pH and enzyme activities) than direct effects. However, the indirect effects of the acid rain S/N ratio (0.265) on the fine root decomposition rate through changes in soil urease activities and the content of litter elements were lower than the pH of acid rain.ConclusionsOur results suggested that the acid rain S/N ratio exacerbates the inhibitory effects of acid rain pH on the decomposition of fine root litter.

The Selective NMDA Receptor GluN2B Subunit Antagonist CP-101,606 with Antidepressant Properties Modulates Cytochrome P450 Expression in the Liver.

Recent research indicates that selective NMDA receptor GluN2B subunit antagonists may become useful for the treatment of major depressive disorders. We aimed to examine in parallel the effect of the selective NMDA receptor GluN2B subunit antagonist CP-101,606 on the pituitary/serum hormone levels and on the regulation of cytochrome P450 in rat liver. CP-101,606 (20 mg/kg ip. for 5 days) decreased the activity of CYP1A, CYP2A, CYP2B, CYP2C11 and CYP3A, but not that of CYP2C6. The alterations in enzymatic activity were accompanied by changes in the CYP protein and mRNA levels. In parallel, a decrease in the pituitary growth hormone-releasing hormone, and in serum growth hormone and corticosterone (but not T3 and T4) concentration was observed. After a 3-week administration period of CP-101,606 less changes were found. A decrease in the CYP3A enzyme activity and protein level was still maintained, though no change in the mRNA level was found. A slight decrease in the serum concentration of corticosterone was also maintained, while GH level returned to the control value. The obtained results imply engagement of the glutamatergic system in the neuroendocrine regulation of cytochrome P450 and potential involvement of drugs acting on NMDA receptors in metabolic drug–drug interactions.

TREATMENT OF ACUTE INTESTINAL INFECTION IN MINKS

The article presents the results of studies on the effectiveness of the multicomponent drug Nifulin (powder for oral administration) in the treatment of acute gastrointestinal infection in minks. The active pharmaceutical ingredients (API) of the drug are metronidazole, furazolidone and oxytetracycline hydrochloride. The researches were conducted on 135 minks of standard dark brown breed at the age of 4–5 months with acute gastrointestinal infection. To isolate and identify pathogens from sick minks with intestinal infection, samples of feces were taken, and from animal’s corpses – samples of parenchymal organs. On the basis of clinical signs of the disease and results of bacteriological researches of biomaterial, a diagnosis – salmonellosis has been put. The test for the sensitivity of bacteria-isolates to oxytetracycline showed a high degree of sensitivity of isolated strains of Salmonella to this antibiotic.
 Nifulin was administered orally at a dose of 0,3g/kg body weight with food three times a day for 3 days. For laboratory tests blood sampling was performed from 10 animals before treatment and on the 10th day from the latest application of the drug.
 Morphological studies of blood samples of sick minks before treatment established a significant increase in the number of leukocytes, and in leukoformula – an increased relative number of rod-nucleated neutrophils and lymphocytes, a decreased number of segmentonuclear neutrophils. Biochemical studies of blood serum revealed a decrease in total protein content, increased transaminase enzyme activity and increased urea and creatinine content, compared with physiological norms.
 On the 10th day after antibiotic therapy hematological studies established a reliable decrease in the relative number of leukocytes by 65,4%, in the number of rod-nucleated neutrophils by 1,3 times and in the number of lymphocytes by 1,9 times, an increase in the number of segmentonuclear neutrophils by 1,4 times. The results of biochemical tests of mink’s serum blood indicated a reliable increase in the total protein content by 18% and increased ALT and AST enzyme activity, compared with the physiological values. The increase in serum AST activity in 1.8 times was noted, that can be explained, as a consequence of the disease, and the influence of antibiotics on the mink’s liver functional state. In this case, a decrease in urea content in 1.3 times and creatinine content – in 1.5 times was determined, the values of these indicators were normalized, indicating the restoration of the physiological state of the mink’s kidneys.
 As a result of the treatment of minks the clinical signs of the disease were gradually disappeared: diarrhea stopped, body temperature and respiratory frequency normalized, general condition has improved. In most animals (130 goals) clinical recovery came. During the application of Nifulin manifestations of adverse reactions or negative effects in the experimental minks were not observed.

Influence of the SOD2 A16V gene polymorphism on alterations of redox markers and erythrocyte membrane fatty acid profiles in patients with multiple chemical sensitivity.

Chronically increased oxidative stress has been reported in patients with multiple chemical sensitivity (MCS). Recently, a single nucleotide polymorphism of the gene coding for mitochondrial superoxide dismutase (SOD2), namely the missense substitution A16V (C47>T) resulting in alteration of SOD2 enzyme activity, has been reported to be associated with MCS. However, the influence of SOD2 A16V genetic background on redox status of patients with MCS has not yet been investigated. Here, the results of a retrospective analysis aimed to evaluate the role of the SOD2 A16V polymorphism in the alterations of antioxidant defense markers as well as fatty acid (FA) composition of erythrocyte membranes in 67 patients with MCS matched with 55 healthy controls is reported. The mutated SOD2 V16 variant was observed more frequently in the MCS group compared with the control group, and this difference was statistically significant. The most common genotype in both groups was the heterozygous SOD2 AV16 variant, whereas the mutated SOD2 VV16 variant was more frequently observed in the MCS group, although the difference was not significant. The MCS cohort showed significantly depleted levels of plasma total antioxidant activity, ubiquinol, erythrocyte reduced glutathione and membrane polyunsaturated FA levels, coupled with significant increases in glutathione peroxidase activity, likely accounting for sustained detoxification from lipoperoxides. Notably, the highest levels of oxidative stress were found in patients with MCS bearing the genotype SOD2 AA16, whereas intermediate levels were found in patients bearing the heterozygous AV16 genotype. Healthy subjects bearing the SOD2 AA16 genotype also showed increased oxidative stress compared with carriers of other SOD2 genotypes. Despite the need for further confirmations in larger cohorts, due to MCS population genetic heterogeneity, these preliminary findings suggest that SOD2 defective activity makes certain patients with MCS more susceptible to developing oxidative stress following a chronic daily exposure to pro-oxidant insults.

Evaluation of prophylactic efficacy of cinnamaldehyde in murine model against Paradendryphiella arenariae mycotoxin tenuazonic acid-induced oxidative stress and organ toxicity

Cinnamaldehyde (Cin) is a natural product obtained from cinnamon and is reported to have a potential anti-fungal, anti-oxidant, anti-inflammatory and anticancer effect. The present study investigated the possible protective role of Cin against tenuazonic acid-induced mycotoxicity in the murine model. Tenuazonic acid (TeA), a toxin produced by Alternaria is a common contaminant in tomato and tomato-based products. Here, Swiss male mice were administered with TeA isolated from Paradendryphiella arenariae (MW504999) (source-tomato) through injection (238 µg/kg BW) and ingestion (475 µg/kg BW) routes for 2 weeks. Thereafter, the prophylaxis groups were treated with Cin (210 mg/kg BW). The experiment was carried out for 8 weeks. The treated groups were compared to the oral and intra-peritoneal experimental groups that received the toxin solely for 8 weeks. Haematological, histopathological and biochemical aspects of the experimental and the control mice were analysed. Sub-chronic intoxication of mice with TeA showed elevated malondialdehyde (MDA), reduced catalase (CAT) and superoxide dismutase (SOD) production; abnormal levels of aspartate transaminase (AST) and alanine transaminase (ALT). Treatment with Cin reversed TeA-induced alterations of antioxidant defense enzyme activities and significantly prevented TeA-induced organ damage. Thus, cinnamaldehyde showed therapeutic effects and toxicity reduction in TeA induced mycotoxicosis.

Chebulagic acid attenuates HFD/streptozotocin induced impaired glucose metabolism and insulin resistance via up regulations of PPAR γ and GLUT 4 in type 2 diabetic rats

The aim of the present study was to evaluate the effects of chebulagic acid on enzymes involved in carbohydrate metabolism in high-fat diet/streptozotocin-induced diabetic rats. Diabetes was induced in albino male Wistar rats by feeding them with a high-fat diet comprising of 84.3% standard laboratory rat chow, 5% lard, 10% yolk powder, 0.2% cholesterol, and 5% bile salt for 2 weeks. After 2 weeks, the animals were kept in an overnight fast and injected with a low dose of streptozotocin (35 mg/kg, dissolved in 0.1 M sodium citrate buffer, pH 4.5). At the end of the experimental periods, the blood glucose, plasma insulin, hemoglobin, glycated hemoglobin levels, homeostatic model of insulin resistance, glycogen content, glycogen synthase, glycogen phosphorylase were measured in high-fat diet and streptozotocin-induced diabetic rats whereas the glucose metabolic enzymes such as hexokinase, glucose 6 phosphate dehydrogenase, glucose 6 phosphatase, fructose 1,6 bisphosphatase and protein expression of peroxisome proliferator-activated receptor–γ (PPAR-γ) and Glucose transporter 4 (GLUT4) were analyzed. Administration of chebulagic acid to HFD/STZ -induced diabetic rats for 30 days significantly decreased the levels of plasma glucose, homeostatic model assessment of insulin resistance, glycogen phosphorylase and glycosylated hemoglobin and increased the levels of insulin, glycogen content and glycogen synthase. On the other hand, the altered activity of carbohydrate metabolic enzymes, PPAR-γ and GLUT4 were brought back to near-normal levels on treatment with chebulagic acid. The effect produced by chebulagic acid on various parameters was comparable to that of metformin.

Deciphering Structural Alterations Associated with Activity Reductions of Genetic Polymorphisms in Cytochrome P450 2A6 Using Molecular Dynamics Simulations.

Cytochrome P450 (CYP) 2A6 is a monooxygenase involved in the metabolism of various endogenous and exogenous chemicals, such as nicotine and therapeutic drugs. The genetic polymorphisms in CYP2A6 are a cause of individual variation in smoking behavior and drug toxicities. The enzymatic activities of the allelic variants of CYP2A6 were analyzed in previous studies. However, the three-dimensional structures of the mutants were not investigated, and the mechanisms underlying activity reduction remain unknown. In this study, to investigate the structural changes involved in the reduction in enzymatic activities, we performed molecular dynamics simulations for ten allelic mutants of CYP2A6. For the calculated wild type structure, no significant structural changes were observed in comparison with the experimental structure. On the other hand, the mutations affected the interaction with heme, substrates, and the redox partner. In CYP2A6.44, a structural change in the substrate access channel was also observed. Those structural effects could explain the alteration of enzymatic activity caused by the mutations. The results of simulations provide useful information regarding the relationship between genotype and phenotype.

Study of some biochemical materials parameters in patients with angina pectoris

In this study, some biochemical data were evaluated in the blood serum of patients with angina pectoris, their ages ranged from (40 to 49) years, and the results were compared with healthy people. Cholesterol and serum enzymatic activity of Aspartate amino transferase AST; Alanine amino transferase ALT; Creatin Kinase Isoenzyme was estimated. Results can be summarized as follow A significant increment (p < 0.05) in the enzymatic activity of AST, ALT and CK-MB enzymes and Cholesterol concentration in patients compared with control group.

PROLIDASE: A Review from Discovery to its Role in Health and Disease.

Prolidase (peptidase D), encoded by the PEPD gene, is a ubiquitously expressed cytosolic metalloproteinase, the only enzyme capable of cleaving imidodipeptides containing C-terminal proline or hydroxyproline. Prolidase catalyzes the rate-limiting step during collagen recycling and is essential in protein metabolism, collagen turnover, and matrix remodeling. Prolidase, therefore plays a crucial role in several physiological processes such as wound healing, inflammation, angiogenesis, cell proliferation, and carcinogenesis. Accordingly, mutations leading to loss of prolidase catalytic activity result in prolidase deficiency a rare autosomal recessive metabolic disorder characterized by defective wound healing. In addition, alterations in prolidase enzyme activity have been documented in numerous pathological conditions, making prolidase a useful biochemical marker to measure disease severity. Furthermore, recent studies underscore the importance of a non-enzymatic role of prolidase in cell regulation and infectious disease. This review aims to provide comprehensive information on prolidase, from its discovery to its role in health and disease, while addressing the current knowledge gaps.

Metastases-Like Liver Lesions in Two Different Types of Porphyria - Porphyria Cutanea Tarda (PCT) and Acute Hepatic Porphyria (AHP) - and the Role of CEUS.

Porphyria are a group of metabolic disorders caused by altered activity of enzymes in the heme biosynthesis pathway. Acute hepatic porphyria (AHP) are due to hepatic overproduction of the porphyrin precursors, delta aminolevulinic acid, and porphobilinogen, and the symptoms are probably caused primarily by injury to the nervous system. However, abdominal pain and nausea can be observed.

A molecular dynamics simulations study of the ionic liquid effect on the β-glucosidase active site interactions with a flavonoid glycoside

Since the ionic liquid (IL) media is an appropriate environment for the enzymatic extraction of glycosides from natural resources, a detailed molecular understanding of the effect of IL concentration on enzyme function, with a focus on enzyme-substrate interactions, is critical. The best docking pose of Quercetin-4′-O-beta-D-glucopyranoside (Q-4′) in the active site of the β-glucosidase (BGL) enzyme was determined using molecular docking methods. Since flavonoid glycosides and enzymes are soluble in imidazolium-based ILs, the aqueous 1-butyl-3-methylimidazolium chloride ([BMIM][Cl]) was used as a solvent in this study. The interactive residues of the BGL active site, as well as the types of interactions, in BGL–Q-4′ complex were then discovered using structural interaction fingerprints (SIFt) calculations. After that, the molecular perspective of the ionic liquid (IL) concentration effect on this complex and its interactions were simulated using 100 ns all-atom MD simulations. According to SIFt changes, changing the IL concentration influenced the overall interactions in the BGL–Q-4′ complex and involved residues. The RMSF findings indicated that the maximum flexibility of the interactive residues was observed in the 50% IL solution, however, flexibility changes were not high enough to alter enzyme activity. More analysis showed that solvent effects influenced hydrogen bond (HB) interactions with the substrate in both active site subsites; glycone (subsite -1) and aglycone subsites. Aromatic interactions followed a similar trend, suggesting that changes in IL concentration affected them. The solvent effects had a greater impact on the interactions with the aglycone part in both cases, which was attributed to aglycone subsite's greater solvent accessibility. Furthermore, it was found that IL concentration had a negative impact on the enzymatic hydrolysis process by influencing the formation of HB with Glu193 as well as the nucleophilic attack of Glu402. Finally, SDF findings indicated that an accumulation of IL molecules at the active site entrance could reduce enzyme activity by reducing the active site's accessibility to the substrate. By providing a molecular perspective on the BGL-substrate complex behavior in the IL system, our findings lead to a deeper understanding of the flavonoid glycoside hydrolysis process in these useful solvents.

Effect of round-shaped silver nanoparticles on the genetic and functional diversity of soil microbial community in soil and “soil-plant” systems

Silver and silver nanoparticles (AgNPs) are extensively used as antimicrobial agents in various products and consequently enter the soil ecosystem, in which they accumulate and can have adverse effects on above and belowground organisms. Since the composition of the soil microbiome could have a significant impact on soil fertility, we focused, in this study, on the effect of AgNPs on soil microbial communities. We evaluated the biomass, plant growth-promoting activities, and microbial genetic (taxonomical) and functional diversity in the soil and the “soil-plant” system using chemical, microbiological, and molecular biological methods, especially qPCR, RFLP, and REP-PCR. By compiling the results of the applied assays, we were able to correlate the changes in genetic diversity with the changes in functional diversity and their impact on the composition of soil organic matter. The effect of AgNPs was predominantly dose-dependent, whereas the concentration 100 mg kgdw−1 had the highest effect on all followed parameters. From the taxonomical point of view, AgNPs had the most significant impact on the relative abundance of Bacteroidetes, which increased in a dose-dependent manner, and Firmicutes, which decreased in a dose-dependent manner, and they caused the predominance of Ascomycota among Fungi. These changes in taxonomic diversity result in a significant decrease in the functional diversity of the microbial communities. Especially for the concentration 100 mg kgdw−1, we detected a significant decrease in the ability of the microbial community to utilize simple carbon substrates, determined by Biolog EcoPlates. We detected also a significant alteration in enzyme activity. The activity of enzymes responsible for depolymerization of complex organic matter was in a dose-dependent manner increased, whereas the highest activity reach samples amended with AgNPs at 100 mg kgdw−1. All these changes result in the accumulation of simple organic matter, which could increase osmotic pressure and have severe consequences on soil fertility. In amended soils, we detected alterations of activity of plant growth-promoting bacteria, specifically in the production of indole-3-acetic acid, 1-aminocyclopropane-1-carboxylic acid deaminase, and siderophores. All these activities are related to mitigating abiotic stress. The presence of plants in experimental soil had both a mitigating and deepening effect on the toxicity of AgNPs. Both effects probably relate to the influence of AgNPs on the selection of different microbial communities in the soil and the “soil-plant” system.

Effect of Short-Term Cold Treatment on Carbohydrate Metabolism in Potato Leaves.

Plants are often challenged by an array of unfavorable environmental conditions. During cold exposure, many changes occur that include, for example, the stabilization of cell membranes, alterations in gene expression and enzyme activities, as well as the accumulation of metabolites. In the presented study, the carbohydrate metabolism was analyzed in the very early response of plants to a low temperature (2 °C) in the leaves of 5-week-old potato plants of the Russet Burbank cultivar during the first 12 h of cold treatment (2 h dark and 10 h light). First, some plant stress indicators were examined and it was shown that short-term cold exposure did not significantly affect the relative water content and chlorophyll content (only after 12 h), but caused an increase in malondialdehyde concentration and a decrease in the expression of NDA1, a homolog of the NADH dehydrogenase gene. In addition, it was shown that the content of transitory starch increased transiently in the very early phase of the plant response (3–6 h) to cold treatment, and then its decrease was observed after 12 h. In contrast, soluble sugars such as glucose and fructose were significantly increased only at the end of the light period, where a decrease in sucrose content was observed. The availability of the monosaccharides at constitutively high levels, regardless of the temperature, may delay the response to cold, involving amylolytic starch degradation in chloroplasts. The decrease in starch content, observed in leaves after 12 h of cold exposure, was preceded by a dramatic increase in the transcript levels of the key enzymes of starch degradation initiation, the α-glucan, water dikinase (GWD-EC 2.7.9.4) and the phosphoglucan, water dikinase (PWD-EC 2.7.9.5). The gene expression of both dikinases peaked at 9 h of cold exposure, as analyzed by real-time PCR. Moreover, enhanced activities of the acid invertase as well as of both glucan phosphorylases during exposure to a chilling temperature were observed. However, it was also noticed that during the light phase, there was a general increase in glucan phosphorylase activities for both control and cold-stressed plants irrespective of the temperature. In conclusion, a short-term cold treatment alters the carbohydrate metabolism in the leaves of potato, which leads to an increase in the content of soluble sugars.

Therapeutic effect of Schisandrin A on avian colibacillosis through gut-liver axis

This study evaluated the therapeutic efficacy of Schisandrin A on systemic colibacillosis of chickens. One hundred and eighty, 1-day-old Hailan Brown chickens were divided into 6 groups of 30 chickens each and assigned to the following treatments: 1) uninfected/untreated control; 2) infected Escherichia coli; 3) infected-plus low dose of Schisandrin A therapy (50 mg/kg); 4) infected-plus medium dose of Schisandrin A therapy (100 mg/kg); 5) infected-plus high dose of Schisandrin A therapy (200 mg/kg) and 6) infected-plus antimicrobial therapy (florfenicol). Each group of chickens was placed in cages with a photoperiod of 12 h of light and 12 h of dark. Feed and water for all groups were provided ad libitum for the duration of the study. On d 14, all the chickens except the uninfected control group were intraperitoneally inoculated with a fresh culture of E. coli containing 1 × 108 CFU/mL. The parameters measured included: average daily weight gain (ADG), percent survivability, liver index, serum activity of enzymes (ALT and AST), hepatic and intestinal concentrations of TNF-α, IL-1β, IL-6, IL-8, and LPS, expression of tight junction proteins (occludin, ZO-1, and claudin-1), relative abundance of bacterial species and histopathological changes in hepatic and intestinal tissue. The results showed that the medium and high doses of Schisandrin A ameliorated the detrimental effects of colibacillosis on weight gain. Regarding organ indexes, E. coli infection induced a significant increase in liver index, all the doses of Schisandrin A produced a significant reduction of liver index in comparison to the E. coli infected control. Serum activity of ALT and AST enzymes significantly increased due to E. coli infection, with the exception of the low dose of Schisandrin A for AST enzyme activity, all the Schisandrin A treatments significantly lowered enzyme activity in comparison to the E. coli infected control. Regarding concentrations of inflammatory markers in hepatic and intestinal, E. coli infection caused a significant increase in TNF-α, IL-1β, IL-6, and IL-8, except the lowest dose of Schisandrin A for IL-1β, the rest of the doses tested were able to significantly reduced the concentrations of inflammatory markers. Concentrations of LPS in hepatic and intestinal tissues were significantly increased by E. coli infection, all doses of Schisandrin A significantly reduced the concentration of LPS in hepatic and intestinal tissue. E. coli infection significantly reduced the expression of 2 tight junction proteins (ZO-1 and Claudin-1), the higher doses of Schisandrin A were effective in significantly increasing the expression of these tight junction proteins when compared with the E. coli infected control. Taken together, these results show that Schisandrin A has potential as an alternative therapy for the treatment of colibacillosis in chickens.

Pectinolytic activity of aspergillus niger and Aspergillus flavus grown on grapefruit (citrus Parasidis) peel in solid state fermentation

The present study was aimed at studying pectinolytic activity of resident fungi isolated from decomposing grapefruit (Citrus parasidis) peels in solid state fermentation. Grape fruit peel was subjected to natural fermentation and the fermenting fungi were isolated, characterized and identified using standard microbiological methods. The isolated fungi were in turn used for fermentation to determine their pectinolytic activity through solid state fermentation technique. Culture parameters such as incubation period, temperature, moisture content and addition of salts supplements were optimized during the research for five days. The identified fungi were Aspergillus Niger and Aspergillus flavus. The peak of pectinolytic activity was at day three of fermentation when the highest pectinase activity of 13.32 μmol/mg/min was recorded for A. Niger and 11.32 μmol/mg/min for A. flavus. Optimum temperature and pH for pectinase activity by A. Niger and A. flavus was at 40 0C and pH 7.5 and 7.7 respectively. The use of salt supplemented substrate did not alter enzyme activity. In conclusion, the isolated fungi could be promising organisms for pectinolytic enzyme production on grape peel as substrate.&#x0D; Keywords: Grapefruit, Pectinolytic Activity, Fungi, Fermentation, Aspergillus

Experience of measuring glutathione peroxidase activity in surgically induced endometrial-like lesions in rats

BACKGROUND: Endometriosis is known to be linked with altered activities of antioxidant enzymes and with their gene polymorphisms. Progestins are known to induce glutathione peroxidase activity in the endometrium and promote reduction of endometrial lesions. It could be useful to estimate the correlation between the activity of glutathione peroxidase within endometrial lesions and their degree of reduction.&#x0D; AIM: The present study was aimed at estimating glutathione peroxidase activity in surgically induced endometrial-like lesions of different degree of reduction in rat model of endometriosis.&#x0D; MATERIALS AND METHODS: The method for determining glutathione peroxidase activity using hydrogen peroxide as a substrate and 5,5-dithiobis(2-nitrobenzoic acid) for estimation of residual reduced glutathione was applied for quantitative analysis of the enzyme activity in endometriotic foci, surgically induced in female Wistar rats. An assay of glutathione peroxidase activity in tissue homogenates was performed at 37C in a reaction medium containing Tris-HCl buffer supplemented with tetrasodium ethylenediaminetetraacetate and sodium azide (pH 8.5) in the presence of 0.55 mM reduced glutathione and 0.192 mM hydrogen peroxide. Before adding trichloroacetic acid, 40-second incubation was used. The correlation between the specific activity of the enzyme and protein amount in endometriotic foci was estimated.&#x0D; RESULTS: In a rat model of endometriosis, there was a high, well-determined glutathione peroxidase activity in endometriotic foci. For the same endometriotic tissue sample, the enzymatic activity was proportional to the amount of protein in the reaction mixture. The range of specific glutathione peroxidase activity was 2.436.45 micromoles of consumed glutathione per minute per milligram of protein (n = 7). In most reduced endometriotic foci (with the minimum amount of endometriotic tissue), the highest specific activity of glutathione peroxidase was found (the Spearmans rho of 0.93 with p = 0.0067).&#x0D; CONCLUSIONS: The method for determining glutathione peroxidase activity using hydrogen peroxide and 5,5-dithiobis(2-nitrobenzoic acid) is convenient for working with the endometriotic tissue in a rat model of endometriosis. We can accept, with p 0.01, that weight of endometriotic foci is negatively linked with specific glutathione peroxidase activity within their tissue. The results are analogous to the previously obtained data on catalase activity and suggest the involvement of both antioxidant enzymes in reduction of endometrial lesions.