Changes In Enzymes Research Articles

Tailoring surface properties of functionalized graphene papers aiming to enzyme immobilization

The use of enzymes as catalysts requires recovery and reuse to make the process viable. Enzymatic immobilization changes enzyme stability, activity, and specificity. It is very important to explore new substrates for immobilization with appropriate composition and structure to improve the efficiency of the immobilized enzymes. This work explores the use of two different graphene oxide papers, one produced by oxidation route (GO) and the other by electrochemical synthesis (EG), aiming for β-galactosidase immobilization. The chemical and structural properties of these two papers were characterized by Raman spectroscopy, X-ray photoelectron spectroscopy and X-ray diffraction. Atomic force microscopy images showed that EG paper ensured more efficient immobilization of the enzymes on the surface of the paper. Cyclic voltammetry was used to monitor the reaction of conversion of lactose to glucose in the free enzyme solution and graphene paper immobilized enzyme solutions. The cyclic voltammetry analysis showed that immobilized enzymes on GO paper showed an improvement in the activity of β-galactose when compared to free enzyme solution, as well as enzyme immobilized on a glassy carbon electrode.

UV-C treatment inhibits browning, inactivates Pseudomonas tolaasii and reduces associated chemical and enzymatic changes of button mushrooms.

Button mushrooms with completely white appearance are popular with consumers. However, button mushrooms are susceptible to infection with Pseudomonas tolaasii, which results in browning. This study evaluates the effects of ultraviolet-C (UV-C) treatment on the inactivation of P. tolaasii in vitro and in vivo and on the physiological and chemical changes of button mushrooms during storage for 21 days at 4 °C. UV-C doses of 0.5 to 9.0 kJ m-2 resulted in 3.91-6.26 log CFU mL-1 reduction of P. tolaasii populations in vitro, and UV-C treatment reduced P. tolaasii populations inoculated on mushroom cap surfaces and browning severity. Moreover, P. tolaasii increased polyphenol oxidase (PPO) activity, and decreased phenylalanine ammonia-lyase (PAL) activity, the accumulation of phenolics and contents of brown melanin precursors, including γ-l-glutaminyl-4-hydroxybenzene (GHB), γ-l-glutaminyl-3,4-dihydroxybenzene (GDHB), and tyrosine in button mushrooms. UV-C treatment was found to reduce the negative changes due to P. tolaasii infection. These results indicated that the application of UV-C treatment inhibited browning, inactivated P. tolaasii and reduced P. tolaasii - associated chemical and enzymatic changes of button mushrooms. © 2021 Society of Chemical Industry.

Anorexia nervosa and pancreatitis.

Acute pancreatitis is a condition whereby erroneous activation of trypsin and zymogen results in pancreatic autodigestion. There are many aetiologies, with alcohol intake and gallstones being the most common. Anorexia nervosa is an eating disorder in which patients' reduced food intake and psychological aversion of weight gain can result in low body weight and malnourishment. The link between pancreatitis and anorexia nervosa is not well understood; this article explores the theorised pathophysiology connecting the two conditions, as well as the optimal management of patients when the conditions co-exist based on current literature. A literature search was performed using MEDLINE, EMBASE and CINAHL databases for all journal articles on the topic of presentations of acute or chronic pancreatitis in adults with anorexia nervosa. The literature proposes various links between anorexia nervosa and pancreatitis. It is theorised that pancreatitis may arise as a result of malnourishment itself or secondary to the refeeding process. Some explanations focus on the histopathological changes to the pancreas that malnourishment induces, while others focus on the enzymatic changes and oxidative damage that arise in the malnourished state. More mechanical mechanisms such as gastric dilatation, gastrointestinal ileus and compartmental fluid shift during refeeding have also been proposed as explanations for the link between the conditions. Some medications used in the management of anorexia nervosa have also been linked to pancreatitis.

Enzymatic Effectiveness of Alcoholic and Aqueous Extract of Salvia Officinalis in Mice Poisoned with Tetrachloride.

Regarding the antioxidant, anti-inflammatory, and antibacterial effects of Salvia officnalis (S. officinalis) extracts and the use of medicinal herbs as an alternative to chemical drugs, this study aimed to evaluate the enzymatic changes and reduction of hepatocyte damage in mice poisoned with carbon tetrachloride (CCl4) after treatment with aqueous and alcoholic extract of Salvia officnalis. A total of 40 adult male mice were divided into eight groups including six experimental, one negative, and one positive control group, which were exposed to CCl4 at the concentration of 2.3 mg/kg. The active compounds in the alcoholic and aqueous extracts of S. officinalis were obtained using high-performance liquid chromatography. Subsequently, S. officinalis extract in 100, 200, and 300 mg /kg doses were fed orally to mice for six days. The enzymes (GST, ALP, ALT, AST, and MDA) were determined in mice serum. The study results showed that enzyme activity was significantly decreased in the group treated with S. officinalis extract, and the concentration of 300mg/kg proved to be most effective. In addition, it was indicated that the alcoholic extract had a higher effect than the aqueous extract, which might be due to the greater amount of active compounds in the alcoholic extract. The improving effects of S. officinalis can be attributed to the bioactive components with antioxidant properties that inhibit the damaging effects of free radicals, chemical drugs, and tissue damage.

Haemato-biochemical and ionic regulatory responses of the hybrid catfish, Heteroclarias, to sublethal concentrations of palm oil mill effluents

BackgroundPollution arising from the noxiousness of palm oil mill effluent (POME) has become a serious threat to aquatic biotas. However, a paucity of information exists on fish response to POME-mediated oxidative stress, lipid peroxidation, haemato-biochemical, enzymatic and ionic changes. This study, therefore, evaluates the effects of 28-day exposure of Heteroclarias, a hybrid catfish, to POME. Juvenile Heteroclarias (n = 350, average weight: 11.90 ± 0.70 g and average length: 9.04 ± 0.71 cm) were exposed to sublethal concentrations (0-control, 4.00, 8.00 and 12.00 mg/l) of POME to determine its effects on red blood cells (RBC), haemoglobin (HB), packed cell volume (PCV), white blood cells (WBC), mean cell haemoglobin (MCH), mean cell haemoglobin concentration (MCHC) and mean cell volume (MCV). The activities of alanine aminotransaminase (ALT), aspartate aminotransaminase (AST), lactate dehydrogenase (LDH), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and the levels of malondialdehyde (MDA), calcium (Ca+), potassium (K+), magnesium (Mg2+) and sodium (Na+) ions were also assayed in the gill and liver.ResultsExposure to POME caused significant (p < 0.05) concentration-dependent decrease in RBC, HB, PCV, MCV, MCH and MCHC values, whereas a significant (p < 0.05) concentration-dependent increase in WBC was observed in POME-exposed fish compared to the control. ALT, AST, LDH, GPx, SOD and CAT activities in the gill and liver significantly (p < 0.05) increased in a concentration-dependent manner. MDA level significantly (p < 0.05) increased in the gill relative to control, while in the liver it was insignificantly different. Both tissues exhibited an increase in Ca+, K+, Mg2+ and Na+ levels at the highest toxicant concentration with a rise of 77.93, 38.46, 109.54 and 41.99% recorded for the electrolytes in the gill and 79.17, 26.92, 55.48 and 38.78% in the liver above the control value, respectively. The levels of all the electrolytes except K+ were higher in the gill than the liver and were in the order: Na+ > K+ > Mg+ > Ca2+ in both tissues.ConclusionsThese results may be used as a suitable tool for pollution assessment and policy formulation to mitigate the discharge of untreated POME into aquatic ecosystems and their impacts on resident organisms.

Enzymatic changes during lethal and sublethal exposures of Glyphosate (41% SL) on freshwater fish Cirrhinus mrigala

The lethal and sublethal effect of the Glyphosate (41% SL) on certain enzymes in tissue such as AAT (Aspartate Amino Transferase), ALAT (Alanine Amino Transferase), LDH (Lactate Dehydrogenase) and ACP (Acid Phosphatase) of the freshwater fish, Cirrhinus mrigala during and after the cessation of the exposure were observed. Ten fish were exposed chronically to lethal and sublethal concentrations for a period of 4 days were sacrificed and tissues such as brain, gill, liver, kidney and muscle were analysed for biochemical parameters and remaining five fish were maintained in clean well water for a period of seven days without test substance being spilled in the water and its tissues were collected after reversal period for biochemical analysis. Marked changes were observed in the biochemical parameters of the fish exposed and after the cessation of the exposure

Ankrd31 in Sperm and Epididymal Integrity.

Ankyrin proteins (ANKRD) are key mediators linking membrane and sub-membranous cytoskeletal proteins. Recent findings have highlighted a new role of ANKRD31 during spermatogenesis, elucidating its involvement in meiotic recombination and male germ cell progression. Following testicular differentiation, spermatozoa (SPZ) enter into the epididymis, where they undergo several biochemical and enzymatic changes. The epididymal epithelium is characterized by cell-to-cell junctions that are able to form the blood-epididymal barrier (BEB). This intricate epithelial structure provides the optimal microenvironment needed for epididymal sperm maturation. To date, no notions have been reported regarding a putative role of ANKRD31 in correct BEB formation. In our work, we generated an Ankrd31 knockout male mouse model (Ankrd31–/–) and characterized its reproductive phenotype. Ankrd31–/– mice were infertile and exhibited oligo-astheno-teratozoospermia (a low number of immotile SPZ with abnormal morphological features). In addition, a complete deregulation of BEB was found in Ankrd31–/–, due to cell-to-cell junction anomalies. In order to suggest that BEB deregulation may depend on Ankrd31 gene deletion, we showed the physical interaction among ANKRD31 and some epithelial junction proteins in wild-type (WT) epididymides. In conclusion, the current work shows a key role of ANKRD31 in the control of germ cell progression as well as sperm and epididymal integrity.

Antioxidant enzymatic changes in bell pepper fruit associated with chilling injury tolerance induced by hot water.

Green bell pepper is highly susceptible to low temperature. The activation of the enzymatic antioxidant system plays a determining role in tolerance to chilling injury (CI). Immersion in hot water for short time previous to storage at low temperature induces tolerance to this disorder. However, there is a lack of information about the induction of chilling tolerance in bell pepper by hot water and its relationship with the enzymatic antioxidant system. We evaluated the effect of three immersion times (T, 1-, 2-, 3-min) in hot water (HW, 53°C) on the reduction of CI in bell pepper and its relationship with the enzymatic antioxidant system during storage at 5°C and 21°C. The use of hot water for 1-, 2- or 3-min reduced the decay and CI indexes, maintained quality parameters, ascorbic acid, and total phenolics content. The storage at 5°C by itself activated the enzymatic antioxidant system. The use of HWT 1-, 2-, and 3-min helped to increase this effect, especially by HWT2 . PRACTICAL APPLICATIONS: The application of a treatment with hot water for short times in fruit sensitive to chilling injury is undoubtedly a viable alternative to increase their tolerance and commercialization. In this study, the application of a hot water treatment for 1-, 2- or 3-min in bell pepper reduced the deterioration and susceptibility to chilling injury and stimulated the enzymatic antioxidant system. In this sense, agricultural producers can take advantage of this treatment to prolong the storage period of the fruit maintaining its quality and improving its commercialization.

A New Method for Exonuclease Activity Analysis of Apurinic/Apyrimidinic Endonuclease 1 and Application in Heavy-polluted Ramie

Apurinic/apyrimidinic endonuclease 1 (APE1) exonuclease plays a vital proofreading role in the base excision repair pathway by removing 3’end group, including oxidatively damaged DNA bases, chain terminating nucleotide analog drugs, terminal blocking groups and mismatched bases. Herein, a simple and real time fluorometric method was developed for detecting APE1 exonuclease activity using molecular beacon as substrate. Experimental results demonstrated that the detection limit of this method was 0.01 U/μL for APE1, and the K m value was (0.27 ± 0.10) μmol/L. Moreover, the method was used to screen antibiotics and the results showed that streptomycin sulfate and gentamycin sulfate had a significant inhibitory effect on APE1 exonuclease activity in a concentration-dependent manner. Meanwhile, the assay was used for investigating the inhibitory effects of heavy metal ions on APE1 exonuclease activity. Finally, the method was further used to monitor the effect of heavy metal ions treatment of the APE1 exonuclease activity of ramie, and the results indicated that APE1 exonuclease activity was inhibited in a concentration-dependent manner after treatment with Pb 2+ and Cd 2+ , which was consistent with heavy metal ions assay in vitro . In summary, this method provided an alternative tool for the biochemical analysis for APE1 exonuclease activity, which was hopeful for the assessment for heavy metal ions adsorption according to the enzymatic activity change in phytoremediation. A convenient and sensitive real-time fluorometric method was proposed for APE1 exonuclease activity detection. This method performed well when it was applied for monitoring the effect of Pb 2+ and Cd 2+ treatment on APE1 exonuclease activity of ramie plant. This study provides an alternative tool for the biochemical analysis for APE1 exonuclease activity, which is hopeful for the assessment for heavy metals adsorption according to the enzymatic activity change in phytoremediation.

Nutritional functions and antioxidative enzymes in juice extract from two different maturity stages of low temperature stored phalsa (Grewia subinaequalis D.C.) fruit

Phalsa is an important berry fruit in Asia, but little is known of its postharvest characteristics and especially changes in fruit composition during storage. Investigations were made for nutritional and antioxidative enzymatic changes in the juice extracted from two different maturity stages (M1: pink to red or M2: dark red to purple) during low temperature storage for 0, 2, 4, 6, 8 and 10 days at 5 ± 1 °C and 65 ± 5% RH. The fruit showed significant gradual declines in ascorbic acid (Vitamin C), total phenolics (TPC), total antioxidant capacity (TAC), total carotenoids, anthocyanins, moisture%, crude protein, crude fiber and fat content and the activity of superoxide dismutase (SOD) during storage. However, the total soluble solids (TSS), pH, ash content, and the activity of catalase (CAT) and peroxidase (POD) increased significantly. The polynomial regression analysis indicated that these changes were either quadratic or linear; and the Pearson's correlation among the studied parameters was significant. More mature fruit had higher concentrations of TSS, ascorbic acid, TPC, TAC, total carotenoids and anthocyanins, more juice and less weight-loss compared with less mature fruit. Overall, significant nutritive functions and antioxidative enzymes were found in both maturity stages of phalsa fruit, which experienced significant changes during postharvest storage.

APPLICATION OF MICROCOLUMN HPLC-UV FOR RAPID QUANTITATIVE ANALYSIS OF ARBUTIN, BERGEN-IN AND GALLIC ACID IN BERGENIA CRASSIFOLIA

This work is aimed at the development of a microcolumn HPLC-UV assay for the rapid quantitative analysis of arbutin, bergenin, and gallic acid in Bergenia crassifolia. The results obtained indicate appropriate metrological parameters of the developed assay. It was found that the known methods of quantitative analysis of phenolic glycosides using SPE-spectrophotometry cannot be characterized as selective and accurate, due to the fact that the presences of the impurity compounds that do not belong to the group of phenolic glycosides negatively influenced the results. The developed assay was used for quantitative analysis of wild and commercial samples of B. crassifolia raw material. It was found that the content of arbutin, bergenin, and gallic acid in samples of B. crassifolia rhizomes collected in the Republic of Buryatia was 38.58–45.97, 66.74–139.76 and 1.22–1.65 mg/g, respectively, and for commercial batches of raw materials 20.57–41.37, 35.04–83.94 and 0.22–1.28 mg/g, respectively. It was found that the process of gradual enzymatic changes in the color of B. crassifolia leaves (green, red, black) leads to significant changes in the chemical composition. The most pronounced phenomenon is the degradation of bergenin, the presence of which is noted only in green leaves. The concentration of gallic acid is reduced in black leaves. Arbutin is characterized by an increased content in red leaves (102.02 mg/g). Additionally, a quantitative analysis of the peduncles and flowers of B. crassifolia was realized, and it was shown that they are distinguished by a high content of arbutin 48.40 and 42.15 mg/g, respectively, as well as bergenin in flowers (16.89 mg/g). The study demonstrated that the developed technique can be applied for a quick, selective, and accurate quantitative analysis of three compounds in various organs of B. crassifolia.

Inhibitors of the sodium-glucose transporter type 2 and new possibilities for managing vascular age in patients with type 2 diabetes mellitus

The article discusses the pathophysiological mechanisms of the development of vascular aging as a combination of the influence on the body of genetic, environmental, regulatory, metabolic and other factors causing biochemical, enzymatic and cellular changes in the arterial vascular bed. The concept of “early vascular aging” and “healthy vascular aging” is defined depending on the ratio of the biological and chronological age of the vessels. The role of diabetes mellitus in increasing vascular stiffness, early vascular aging, as well as the progression of atherosclerotic cardiovascular diseases and their complications is considered in detail. Approaches to multifactorial management of vascular age in patients with type 2 diabetes (lifestyle modification with strategy of aggressive treatment of modifiers of atherosclerosis, rejection of bad habits, adherence to dietary recommendations and the use of modern organo- and vasoprotective antidiabetic drugs) are revealed. The mechanism of realization of vasoprotective effects of inhibitors of sodium-glucose transporter-2 (iNGLT-2) is described in detail. The results of completed large random ized trials EMPA-REG Outcome and EMPA-REG BP of the most studied representative of the IGLT-2 group, empagliflozin, are presented. It has been shown that due to their glucose and natriuretic effects, the ability to reduce body weight and blood pressure, improve myocardial metabolism and bioenergetics, decrease the activity of the sympathetic nervous system, as well as positive effects on vascular stiffness, NGLT-2 inhibitors are the drugs of choice in patients with type 2 diabetes mellitus (T2DM) and cardiovascular diseases. This makes it possible to widely use this group of drugs for managing the vascular age of patients and represents a new opportunity in the prevention of vascular aging in T2DM.

The effect of global change on soil phosphatase activity.

Soil phosphatase enzymes are produced by plant roots and microorganisms and play a key role in the cycling of phosphorus (P), an often-limiting element in terrestrial ecosystems. The production of these enzymes in soil is the most important biological strategy for acquiring phosphate ions from organic molecules. Previous works showed how soil potential phosphatase activity is mainly driven by climatic conditions and soil nitrogen (N) and carbon. Nonetheless, future trends of the activity of these enzymes under global change remain little known. We investigated the influence of some of the main drivers of change on soil phosphatase activity using a meta-analysis of results from 97 published studies. Our database included a compilation of N and P fertilization experiments, manipulation experiments with increased atmospheric CO2 concentration, warming, and drought, and studies comparing invaded and non-invaded ecosystems. Our results indicate that N fertilization leads to higher phosphatase activity, whereas P fertilization has the opposite effect. The rise of atmospheric CO2 levels or the arrival of invasive species also exhibits positive response ratios on the activity of soil phosphatases. However, the occurrence of recurrent drought episodes decreases the activity of soil phosphatases. Our analysis did not reveal statistically significant effects of warming on soil phosphatase activity. In general, soil enzymatic changes in the reviewed experiments depended on the initial nutrient and water status of the ecosystems. The observed patterns evidence that future soil phosphatase activity will not only depend on present-day soil conditions but also on potential compensations or amplifications among the different drivers of global change. The responses of soil phosphatases to the global change drivers reported in this study and the consideration of cost-benefit approaches based on the connection of the P and N cycle will be useful for a better estimation of phosphatase production in carbon (C)-N-P models.

Structure and Applications of Pectin in Food, Biomedical, and Pharmaceutical Industry: A Review

Pectin is a biocompatible polysaccharide with intrinsic biological activity, which may exhibit different structures depending on its source or extraction method. The extraction of pectin from various industrial by-products presents itself as a green option for the valorization of agro-industrial residues by producing a high commercial value product. Pectin is susceptible to physical, chemical, and/or enzymatic changes. The numerous functional groups present in its structure can stimulate different functionalities, and certain modifications can enable pectin for countless applications in food, agriculture, drugs, and biomedicine. It is currently a trend to use pectin to produce edible coating to protect foodstuff, antimicrobial bio-based films, nanoparticles, healing agents, and cancer treatment. Advances in methodology, use of different sources of extraction, and knowledge about structural modification have significantly expanded the properties, yields, and applications of this polysaccharide. Recently, structurally modified pectin has shown better functional properties and bioactivities than the native one. In addition, pectin can be used in conjunction with a wide variety of biopolymers with differentiated properties and specific functionalities. In this context, this review presents the structural characteristics and properties of pectin and information on the modification of this polysaccharide, its respective applications, perspectives, and future challenges.

Different mechanisms underlying the divergent responses of soil respiration components to an introduction of N2-fixer tree species into Eucalyptus plantations

Soil respiration (Rs), primarily rhizospheric (Rr) and heterotrophic respiration (Rh) is an important CO2 efflux from soil to the atmosphere. Previous studies demonstrated that introducing N2- fixing species into plantations could increase soil carbon stock and modify carbon chemical composition by altering microbial community and associated enzymes activities. However, their responses of Rs and its two components (Rr and Rh) remain poorly understood. This study investigated how Rs and its components responded to the introduction of N2-fixer tree species in a subtropical eucalyptus plantation and to further ascertain its underlying mechanisms. We measured Rs and its components (Rr and Rh) and used phospholipid fatty acids (PLFAs) as bioindicators to study soil microbial communities and activities of four extracellular enzymes as indicators of soil microbial functioning in an 8-year-old pure Eucalyptus urophylla plantation (PP) and an 8-year-old mixed E. urophylla and Acacia mangium (N2-fixer tree species) plantation (MP) in subtropical China. We found that Rs was significantly reduced by approximately 12.5%, with divergent responses of Rr and Rh in the MP. Compared with PP, the Rr decreased by 41.7% and Rh increased by 36.6% in plantation with N2-fixer species. The decreased Rr in the MP was largely associated with a reduction in fine root biomass. In contrast, the increased Rh in the MP was induced by an increase in total microbial biomass, changing the microbial community structure and enhancing the activities of β-1,4-glucosidase. The enzymatic changes were associated with an increase in N availability. Our results indicate that shifts in both biotic and abiotic factors resulting from the introduction of N2-fixer species shaped changes in Rs and its components.

Abstract 2915: Synergistic, cytokine mediated stimulation (IL-1α/β + IL-6) of colon cancer cells reveals a novel mechanism of DNA damage facilitated by up-regulation of NOX1 and DUOX2

Abstract Inflammatory bowel diseases are states of long-term inflammation of the colon resulting from an interplay of environmental factors and dysregulated immune responses. As inflammatory exposure increases over the time frame of active disease, patients with these chronic conditions are at risk of developing colorectal cancers. Chronic inflammation is thought to contribute to cancer initiation through persistent release of reactive oxygen species (ROS), leading to genome damage. Accumulating evidence suggests inflammatory cells contribute to this process, as the secretion of cytokines and growth factors in response to gut inflammation also supports cancer cell progression. Specifically, IL-1 family members play a pivotal role in the pathogenesis of acute and chronic intestinal inflammation, regulating both innate and adaptive immune responses. To elucidate a direct link between IL-1 family cytokines, ROS generation and colorectal cancer, we observed that treatment of HT29 or T84 cells with IL-1α or IL-1β, in cooperation with IL-6, resulted in oxidant production through up-regulation of both NOX1 and DUOX2 enzymes. Up-regulation of the hydrogen peroxide generating DUOX2/DUOXA2 enzyme complex was directly associated with enhanced histone H2AX phosphorylation (γH2AX), a marker of DNA double strand breaks. Interestingly, this concentration and time-dependent induction of expression and oxidative response was absent in Caco2 cells and was not mediated by other IL-1 family members (IL-18, IL-33 or IL-37). Prior studies from our group have demonstrated significant up-regulation of DUOX2 and DUOXA2 in surgically resected colon cancer specimens compared with adjacent normal colonic epithelium. Future studies will focus on cytokine stimulation of colon organoids derived from patient tumor and adjacent non-malignant tissues to evaluate enzymatic expression and oxidant level changes in a novel patient-derived colon model system. Citation Format: Jennifer L. Meitzler, Yongzhong Wu, Agnes Juhasz, Annamaria Rapisarda, Petreena Campbell, Becky A. Diebold, Smitha Antony, Guojian Jiang, Jiamo Lu, David J. Mallick, Mariam M. Konaté, Krishnendu Roy, James H. Doroshow. Synergistic, cytokine mediated stimulation (IL-1α/β + IL-6) of colon cancer cells reveals a novel mechanism of DNA damage facilitated by up-regulation of NOX1 and DUOX2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2915.

Effects of Olive, Milk Thistle, and Artichoke Extracts on Performance, Biochemical and Enzymatic Changes in Carbon Tetrachloride-intoxicated Broiler Chickens

This study was conducted to examine the effects of olive, milk thistle, and artichoke extracts on growth performance, biochemical and enzymatic changes in birds intoxicated with CCl4 (at 0.5, 0.75, and 1 mL/kg body weight in days 14, 21, and 28 of age, respectively). Two hundred twenty-five 10-day-old Ross 308 female broiler chicks were used in a completely randomized block design to evaluate the effects of five treatments consisting 1) a basal diet given to the birds injected intraperitoneally with 0.5 mL saline (control-), 2) The same basal diet given to birds injected intraperitoneally with CCl4 (control+), 3, 4, and 5) The basal diet supplemented with either 400 mg/kg milk thistle, olive extracts, or artichoke extract given to the birds injected with CCl4. Dietary Supplementation of each herbal extract increased feed intake compared with the control birds (P < 0.05). The broilers fed on the artichoke supplemented diet showed a greater serum concentration of HDL-c than other birds on day 28 of age (P < 0.05). Feeding the control diet to the CCl4 injected birds and the olive extract-included diet with no injection resulted in a greater serum alanine aminotransferase concentration at day 42 of age (P < 0.05). Serum malondialdehyde concentration decreased in the birds given the control diets (P < 0.05). The supplementary olive extract increased serum concentrations of malondialdehyde and total anti-oxidant on day 42 of age (P < 0.05). Liver health score improved in the broilers grown on the herbal extract-added diets (P < 0.05). In conclusion, administration of olive, milk thistle, and artichoke extracts promoted feed intake and exerted a potentially positive role as an antioxidant in liver health as evidenced by certain promising changes in serum biochemistry and enzyme activity as well as liver health score.

Metagenomics and Metabolomics Study on Microbial Diversity and Enzymatic Activities Changes on Cow Dung Based Low-Temperature Biogas Production by Entererococcus spp

To fill the gap between the rise in demand for energy and decline in the traditional energy sources such as coal, natural gas and nuclear energy, other alternative sources such as biogas are necessary. Studies have shown that the existing conditions within the fermentation realm control the microbial characteristics in biogas production. However, there is inadequate insight between the duration of fermentation and the microbial diversity, and with specific emphasis to cow manure as the substrate under low temperature fermentation. This study aimed at providing additional insight on the effect of varying fermentation duration (0 to 60 days) on the composition of the dominant microbial flora on cow dung based low-temperature (15 °C) biogas fermentation using metagenomics and metabolomics analyses approach. The study results showed that the main dominant community in the process of methanogenesis are the Spirochaetae, Synergistetes and Chloroflexi, and are new flora in the methane phase. In the peak stage of low-temperature biogas fermentation, the dominant bacteria groups were Methanosarcina and after adding 10% concentration of L1 bacteria. The prediction of metabolic pathway was mainly carbohydrate metabolism and amino acid metabolism with succinyl-CoA synthase a subunit, lactaldehyde reductase and the glutamate-glyoxylate aminotransferase being the main unique enzymes. The study therefore supports the potential of involving the reported dominant microbial communities and related enzymatic activities for improved biogas production under low temperature conditions.

Evaluation of gingival tissue samples for predicting the time of death using histological and biochemical tests

Thanatochemistry also known as chemistry of death and is used to determine post mortem interval (PMI). It is arguably one of the critical steps in forensic investigation. Recent addition of analyzing biochemical changes along with the traditional methods have gained importance, as they help us to record very early changes in the tissue specimens. In this view, our study aimed to correlate both histological changes and enzymatic changes in gingival tissue samples at intervals of immediate, 1 h, 5 h, 24 h and 48 h after death. Histologic changes noted were loss of epithelial architecture, chromatin clumping, nuclear vacuolation, karryopyknosis, eosinophilia and wide intercellular junctions. Two enzymes which differentiate between the autolytic phase (acid phosphatase) and putrefactive phase (ammonia) of decomposition were evaluated using UV spectrometer. Results in our study demonstrated there were variations as in gradual increase in ammonia levels (1.13±0.24–26.6±2.09) and gradual decrease in acid phosphatase levels (5.61±0.67–1.25±0.53) at different time intervals till 48 h. The cellular changes in gingival tissue could also be related to time. The result of our study helps us to identify potential of enzymatic changes which when correlated with histological reports helps us to predict the time of death accurately. Replicating this experiment in various known taphonomic conditions and other enzymes could highlight the usefulness of gingival tissue samples in determining time of death.

Evaluation of virulence potential of Aspergillus tubingensis and subsequent biochemical and enzymatic defense response of cotton.

Aspergillus tubingensis is a causative known pathogen of various important crops, worldwide. The existing study was aimed to examine the virulence potential of A. tubingensis on resistant (NIA-Sadori) and susceptible (CIM-573) cultivars of cotton. For this purpose, both cultivars were inoculated with pathogen and altered morphology of diseased leaves was observed with light and scanning electron microscope. Disease severity was measured and estimated to be 68.7 and 27.1% in susceptible and resistant cultivars, respectively. To understand and compare defense mechanism of resistant and susceptible cultivars, different biochemical and enzymatic changes were observed. After the infection of A. tubingensis, increase in the concentrations of sugar, total protein, proline, phenol, and phenylalanine ammonia lyase (PAL) was more prominent in resistant cultivar, than the susceptible one. Moreover, due to increased number of dead cells, significantly higher electrolyte leakage was detected in susceptible cultivar. Principal component analysis confirmed the effect of A. tubingensis on growth attributes and various physiological and biochemical activities of cotton. These findings help us to suggest a possible role of proline content, protein content, and PAL activity in resistance mechanism of Cotton plant.