Altered Enzyme Activity Research Articles

Synthetic and biological surfactant effects on freshwater biofilm community composition and metabolic activity

Surfactants are used to control microbial biofilms in industrial and medical settings. Their known toxicity on aquatic biota, and their longevity in the environment, has encouraged research on biodegradable alternatives such as rhamnolipids. While previous research has investigated the effects of biological surfactants on single species biofilms, there remains a lack of information regarding the effects of synthetic and biological surfactants in freshwater ecosystems. We conducted a mesocosm experiment to test how the surfactant sodium dodecyl sulfate (SDS) and the biological surfactant rhamnolipid altered community composition and metabolic activity of freshwater biofilms. Biofilms were cultured in the flumes using lake water from Lake Lunz in Austria, under high (300 ppm) and low (150 ppm) concentrations of either surfactant over a four-week period. Our results show that both surfactants significantly affected microbial diversity. Up to 36% of microbial operational taxonomic units were lost after surfactant exposure. Rhamnolipid exposure also increased the production of the extracellular enzymes, leucine aminopeptidase, and glucosidase, while SDS exposure reduced leucine aminopeptidase and glucosidase. This study demonstrates that exposure of freshwater biofilms to chemical and biological surfactants caused a reduction of microbial diversity and changes in biofilm metabolism, exemplified by shifts in extracellular enzyme activities.Key points• Microbial biofilm diversity decreased significantly after surfactant exposure.• Exposure to either surfactant altered extracellular enzyme activity.• Overall metabolic activity was not altered, suggesting functional redundancy.

Impacts of sub‐optimal water quality on the haemolymph biochemistry, immune status and condition of post‐harvest Southern Rock Lobsters (Jasus edwardsii)

AbstractPalinurid rock (or spiny) lobsters constitute valuable seafood fisheries around the world. Post‐harvest mortality can have a substantial economic impact on the value of these fisheries and damage the brand identity of an otherwise fungible commodity. In this study, sudden, large‐scale mortalities in the Australian Rock Lobster industry were investigated through comprehensive haemolymph biochemistry panels, measurements of immune status and nutritional condition. A cohort of lobsters (n = 25) was sampled over a 4‐week period at a commercial processing facility, with the final sample occurring during a mortality event. A comparison of 22 biochemical parameters, including electrolytes, minerals, ions, metabolites and enzymes, showed a notable disruption in osmoregulation, protein content and enzymes during weeks 3 and 4 of the study. Haemocyte counts showed a decline in circulating haemocytes by the time of the mortality event along with a shift in the proportion of hyalinocytes to granulocytes favouring the former. The condition index was variable over the sampling schedule, possibly linked to observed incidences of cannibalism, with no evidence of an overall low condition that would explain mortalities. The lobsters sampled in this study did not show any consistent or progressive changes in biochemistry that may indicate a pathogen or disease as a causative agent. Rather, analysis of the water quality within the facility suggested that mortality was due to sub‐optimal water quality characterized by a combination of two factors, a substantial decrease in the pH and potential nitrogen toxicity, that caused a physiological response comprised of an ion imbalance, altered enzyme activity and, ultimately, mortality. These results offer a comprehensive characterization of the impact of unexplained morbidity on the biochemistry and physiology of lobsters, illustrate the harm of sub‐optimal aquaculture practices and highlight the need for a better understanding of aquaculture practices and their effects on economically important seafood industries.

Exposure to low-dose arsenic caused teratogenicity and upregulation of proinflammatory cytokines in zebrafish embryos.

Arsenic is currently ranked as the most toxicant on the ATSDR 2015 substance priority list and is categorised as a Group 1 human carcinogen. Biota that are subjected to inorganic arsenicals through food, water, occupational or medical exposure pose a risk to the environment and to human health. The present study was carried out to investigate the toxicity caused by inorganic arsenic. After fertilisation, zebrafish embryos were exposed to sodium arsenite at several concentrations (100nM to 600nM) for 24 to 96 hpf. The indicators of teratogenicity (hatchability, morphological abnormalities, mortality), behavioural modifications (touch induced escape response (TIER), startle response (SR) and turning behaviour (TB)), biochemical testing (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and glutathione S transferase (GST)) and the expressions of tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6) and cyclooxygenase-2 (COX-2) were investigated. The aforementioned parameters were found to be altered in embryos exposed to sodium arsenite. According to the findings of the current study, even a low dose of inorganic arsenic compound caused teratogenicity, behavioural abnormalities, altered enzyme activities and the expression of proinflammatory cytokines in zebrafish embryos.

Short-term response of soil enzyme activities and bacterial communities in black soil to a herbicide mixture: Atrazine and Acetochlor

Combined application of atrazine (ATZ) and acetochlor (ACE) was used as an emerging herbicidal method in black soil zone of northeastern China recently. However, the interaction toxic effect of the two herbicides on the soil biological characteristics has not been investigated yet. In this study, the alterations of soil enzyme activities and bacterial communities in black soil were assayed during a short term (28 days) exposure period by spiking ATZ, ACE or ATZ + ACE (1:1, w: w) at 0×, 1×, 2×, 5× and 10× the recommended dose in a microcosm experiment conducted under greenhouse condition. The results showed that ATZ and ACE had synergistic inhibitory effects on soil enzymes activities. Higher inhibition rates of urease, acid phosphatase, sucrase and catalase were observed in ATZ + ACE, and the enzyme activities could not recover to the level without herbicide application after 28 days. The higher IBRv2 value of ATZ + ACE also indicated that the comprehensive toxicity was stronger than that of ATZ or ACE. The microbial community structure was affected differently by ATZ, ACE and ATZ + ACE as the PLFAs analysis displayed, and ATZ + ACE was more harmful to bacteria than ATZ or ACE. The decrease in relative abundance of Proteobacteria was more sensitive to ATZ, and the decrease of Acidobacteriota was more sensitive to ACE, and the relative abundance of these two phyla decreased more obviously in ATZ + ACE. In addition, the PICRUSt analysis showed that the functions of the replication and repair, energy metabolism, cell motility and cellular processes and signaling were inhibited by ATZ and ACE, especially in the ATZ + ACE treatment. This means that combined application of ATZ and ACE has a greater potential risk on soil microbes and consequently the nutrient cycling process.

EFFECT OF NON-SYNONYMOUS SINGLE-NUCLEOTIDE POLYMORPHISM OF HUMAN CARBOXYL ESTERASE 1 ON THE BIOACTIVATION OF DABIGATRAN ETEXILATE

Objective: Dabigatran Etexilate is an oral acting direct thrombin inhibitor used for prophylaxis of cardioembolic events in non-valvular atrial fibrillation patients. Genetic polymorphisms in hCES1 gene can significantly alter the conformations of the enzyme drug binding and impair the catalytic ability. Hence this study is performed to determine the effect of single nucleotide variations on dabigatran activation by hCES1. Methods: Energy minimization was performing using YASARA server. Dihedral angles of the modeled targets were visualized using Ramachandran Plot. We performed molecular docking analysis with Autodock 4.2. Three-dimensional grid was constructed using Autogrid. Docked complexes were visualized using Pymol Viewer. Results: Significant differences in the binding energy and conformations of Dabigatran in the active site of the target was observed between the Ser75Asn, Ala158Val, Asp203Glu, Ala269Ser and Thr290Met variants. The catalytic triad was completely or partially disrupted in the variants suggestive of altered enzyme activity. Co­nclusion: The binding energy of Dabigatran with the mutant (79Ala, 221Ala, 354Ala and 468Ala) was found to be less than that of the wild type. This indicates that the presence of functional non-synonymous polymorphism in the hCES1 significantly alters the binding of Dabigatran. Hence patients who have this SNP (332G>A, 581C>T, 717C>A, 913C>T, 977C>T) would have decreased hCES1 function, which would result in therapeutic failure or sub-therapeutic drug action.

The acute effect of thiamine on serum insulin levels and some biochemical parameters in excessive alcohol-consuming rats

There are studies that present metabolic disorders in alcohol drinkers are associated with thiamine deficiency. Therefore, in the present study, it was aimed to investigate the effectiveness of thiamine in rats with binge drinking model. For this purpose, total 21 spraque dawley rats were divided into three equal groups as control, alcohol and thiamine+alcohol groups. The thiamine+ethanol group was given thiamine at a daily dose of 100 mg/kg by oral gavage, starting 2 days before the ethanol administration. Alcohol and thiamine+alcohol groups were given 3.45g/kg/day ethanol as 20%. At the end of the study, while serum total bile acid, total bilirubin and insulin levels increased in rats in the alcohol group compared to the rats in the control group; total protein and albumin levels decreased (P<0.05). In the thiamine + alcohol group, LDL-cholesterol, total cholesterol, bile acid levels and AST enzyme activity increased, while ALT enzyme activity and total protein levels decreased compared to the control group (P<0.05). There was no statistically significant result in the values in the thiamine+alcohol group compared to the alcohol group. It has been concluded that acutely administered thiamine supplementation had no effect on alcohol-induced biochemical parameter changes in binge-drinking animals. In this sense, studies with longer-term thiamine use are needed.

Replacement of Trypsin by Proteases for Medical Applications.

Cell culture has a crucial role in many applications in biotechnology. The production of vaccines, recombinant proteins, tissue engineering, and stem cell therapy all need cell culture. Most of these activities needed adherent cells to move, which should be trypsinized several times until received on a large scale. Although trypsin is manufactured from the bovine or porcine pancreas, the problem of contamination by unwanted animal proteins, unwanted immune reactions, or contamination to pathogen reagents is the main problem. This study investigated microbial proteases as a safe alternative for trypsin replacement in cell culture experiments for the detachment of adherent cells. The bacteria were isolated from the leather industry effluent based on their protease enzymes. After sequencing their 16S ribosomal deoxyribonucleic acid, their protease enzymes were purified, and their enzyme activities were assayed. The alteration of enzymatic activities using different substrates and the effect of substrate concentrations on enzyme activities were determined. The purified proteases were evaluated for cell detachment in the L929 fibroblast cells compared to trypsin. The separated cells were cultured again, and cell proliferation was determined by the MTT assay. The results showed that the isolated bacteria were Bacillus pumilus, Stenotrophomonas sp., Klebsiella aerogenes, Stenotrophomonas maltophilia, and Bacillus licheniformis. Among the isolated bacteria, the highest and the lowest protease activity belonged to Stenotrophomonas sp. and K. aerogenes, with 60.34 and 11.09 U/mL protease activity, respectively. All the isolated microbial proteases successfully affected L929 fibroblast cells' surface proteins and detached the cells. A significant induction in cell proliferation was observed in the cells treated with K. aerogenes protease and B. pumilus protease, respectively (P < 0.05). The obtained results suggested that microbial proteases can be used as safe and efficient alternatives to trypsin in cell culture in biopharmaceutical applications.

Qingfei oral liquid alleviates RSV-induced lung inflammation by promoting fatty-acid-dependent M1/M2 macrophage polarization via the Akt signaling pathway

Ethnopharmacological relevanceRespiratory syncytial virus (RSV) is a common pathogen that causes lower respiratory tract disease in infants and the elderly, and no vaccination is presently available. Qingfei oral liquid (QF), a traditional Chinese medicine formula, has been shown in clinic to have anti-inflammatory properties. Aim of the studyThe present study investigated whether QF can suppress RSV-induced lung inflammation in mice models via fatty acid-dependent macrophage polarization. Material and methodsBALB/c mice were given a low, medium, or high dose of QF intragastrically for four consecutive days following RSV infection. The lung inflammatory status was assessed using H&E staining and cytokine assays. The active components of QF and fatty acid metabolism were analyzed using ultra-high-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS). A lipid metabolism-related pathway was found through network pharmacology and molecular docking investigations. Western blotting assays were used to determine the levels of ATP-citrate lyase (ACLY), peroxisome proliferation-activated receptor alpha (PPAR), Akt protein kinase B and its phosphorylated form in Akt signaling. Flow cytometry was used to quantify the number of macrophage subtypes (M1/M2), and immunohistochemistry was used to examine the expression of inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1). ResultsIn the lung tissues of RSV-infected mice, QF suppressed the transcription of pro-inflammatory proteins such as interleukin-1 beta (IL-1β), tumor necrosis factor alpha (TNF-α), and interleukin-6 (IL-6), while increasing the level of anti-inflammatory factors such as interleukin-10 (IL-10). The alterations in metabolic enzyme activity mediated by Akt signaling were linked to QF's significant reduction in lung fatty acid accumulation. Lower ACLY expression and higher PPAR expression were found after QF treatment, showing that these two enzymes were downstream targets of Akt signaling, controlling fatty acid synthesis (FAS) and fatty acid oxidation (FAO), respectively. The reprogramming of fatty acid metabolism resulted in the polarization of macrophages from M1 to M2, with lower expression of iNOS and higher expression of Arg-1. Additionally, application of an Akt agonist (SC-79) reduced QF's anti-inflammatory effects by increasing FAS and decreasing macrophage polarization. ConclusionsQF inhibited Akt-mediated FAS and polarized M1 to M2 macrophages, resulting in an anti-inflammatory impact.

Advanced Strategies of Enzyme Activity Regulation for Biomedical Applications.

Enzymes are important macromolecular biocatalysts that accelerate chemical and biochemical reactions in living organisms. Most human diseases are related to alterations in enzyme activity. Moreover, enzymes are potential therapeutic tools for treating different diseases, such as cancer, infections, and cardiovascular and cerebrovascular diseases. Precise remote enzyme activity regulation provides new opportunities to combat diseases. This review summarizes recent advances in the field of enzyme activity regulation, including reversible and irreversible regulation. It also discusses the mechanisms and approaches for on-demand control of these activities. Furthermore, a range of stimulus-responsive inhibitors, polymers, and nanoparticles for regulating enzyme activity and their prospective biomedical applications are summarized. Finally, the current challenges and future perspectives on enzyme activity regulation are discussed.

Effects of Allanblackia floribunda stem-bark Extracts on Oxidative stress and Mitochondrial Dysfunction in Rats Exposed to Crude Oil

The development of degenerative diseases has been linked to oxidative stress and mitochondrial dysfunction. Additionally, it has been demonstrated that the changes caused by crude oil exposure contribute to the onset of degenerative diseases. There is little or no information on the anti-degeneration properties of Allanblackia floribunda. This study was carried out to investigate brain mitochondrial tissues exposed to crude oil and the effects of A. floribunda as a preventive and therapeutic intervention against oxidative stress and mitochondrial dysfunction. Twenty-five Wistar rats were divided into five groups: the control group received distilled water, the second group received ethanol extracts of A. floribunda, the third group received crude oil (5 mL/kg), the fourth and fifth groups received crude oil and A. floribunda extract concurrently (200 and 400 mg/kg, respectively). The results showed that crude oil treatment caused a significant increase in brain mitochondrial MDA levels and induced significant alterations in brain mitochondrial antioxidant enzyme activities compared with control. Treatment with the extract alone revealed a significant increase in protein carbonyl, conjugated dienes, and ATPases compared with crude oil-treated rats. This indicates that A. floribunda inhibited crude oil-induced mitochondrial lipid peroxidation and improved ATPase and antioxidant status in the rat brain mitochondria, which further suggests that A. floribunda possesses potential and may efficiently inhibit crude oil-induced oxidative damage and improve mitochondrial functions.

Abstract P3041: DWORF Overexpression In The Heart Results In Enhanced Mitochondrial Function

Background: Calcium (Ca 2+ ) plays a pivotal role in regulating physiological processes in the heart including excitation-contraction coupling, Ca 2+ signaling, and mitochondrial function. It is well established that both Ca 2+ dysregulation and altered cardiac energy metabolism are universal features of heart failure (HF). Dwarf open reading frame (DWORF), a newly discovered microprotein, increases SERCA2a activity in part by relieving SERCA2a inhibition by phospholamban (PLN). DWORF overexpression via cardiac specific transgene or gene therapy is cardioprotective in genetic and experimental HF mouse models. In addition to activation of SERCA2a by PLN displacement, preliminary data indicates that enhanced mitochondrial function contributes to the cardioprotective effects of DWORF. Here we tested the effects of DWORF overexpression on cardiac mitochondrial function and Ca 2+ dynamics. Methods and Results: We isolated mitochondria from wildtype (WT) and DWORF transgenic (Tg) mouse hearts and measured O 2 consumption rates using a Seahorse mito stress test. We found that DWORF Tg mice have increased respiratory capacity compared to WT (DWORF Tg 1150.0 pmol/min, WT 799.4 pmol/min +/-146.2) and enhanced spare respiratory capacity (DWORF Tg 489.1 pmol/min, WT 229.4 pmol/min +/- 102.1). To measure mitochondrial Ca 2+ dynamics, we performed Ca 2+ retention capacity assays and found that DWORF Tg mice have increased Ca 2+ uptake rates compared to WT (DWORF Tg 32.32 s, WT 46.46 s +/- 4.39). Western blot analysis of the mitochondrial Ca 2+ uniporter (MCU) in mito extracts from WT and DWORF Tg hearts indicated elevated levels of MCU. However, electron transport chain (ETC) protein expression indicated no significant differences, indicating the enhanced mitochondrial activity observed in DWORF Tg mice was not due to increased ETC levels. Conclusions: DWORF Tg mouse heart mitochondria exhibit increased maximal respiration rates and enhanced spare respiratory capacity. However, ETC protein levels are unchanged, indicating altered enzyme activity rather than expression. Additionally, mitochondria from DWORF Tg hearts uptake Ca 2+ faster into the mitochondria through the increased levels of MCU, suggesting enhanced mitochondrial Ca 2+ dynamics.

Composite biscuits from sandpaper and acha flour restore the altered activity of arginase, cholinergic, and purinergic enzymes in hypertensive-diabetic rats.

Hypertension is one of the common co-morbidities in diabetes. Thus, the present study sought to study the effects of composite biscuits from the mixture of acha (Digitaria exilis) and sandpaper (Fiscus exasperata) leaf flours (ASLF) on mean arterial blood pressure (MABP), arginase, cholinergic, purinergic enzymatic cascade, and nitric oxide (NO) levels as well as oxidative status in streptozotocin (STZ)/L-NG -nitro arginine methyl ester (L-NAME)-induced hypertensive/diabetic rats. Experimental rats were distributed randomly into 7 groups (n=5). Group I-III rats were placed on the basal diet; IV-VII rats were placed on composite biscuits designated as A, B, C, and D respectively for 14 days. On the 13th day, the MABP of the experimental rats was monitored and recorded. Thereafter, the rats were sacrificed, tissues of interest were harvested, and homogenized. Subsequently, the activity of arginase cholinesterase and purinergic enzymes, as well as NO levels were evaluated in the experimental rats. However, hypertensive/diabetic rats placed on the formulated diet exhibited reduced MABP when compared with the untreated hypertensive/diabetic rats. Also, altered activity of arginase, cholinergic and purinergic were restored in diet-treated hypertensive/diabetic rats when compared with hypertensive/diabetic rats. Similarly, the NO level and antioxidant status of the treated hypertensive/diabetic rats were notably enhanced when compared with hypertensive/diabetic rats. It could be inferred that composite biscuits exhibited an ameliorative effect in hypertensive/diabetic states via their reductive effect on the MABP, arginase, cholinesterase, and purinergic enzymes and enhanced NO levels in hypertensive/diabetic rats. Meanwhile, the biscuit designated as D had seems better when their effects were compared holistically. PRACTICAL APPLICATIONS: Acha grains and sandpaper leaf have been used in the folklore for disease treatment. However, the production of composite biscuits from these naturally available recipes for the management of hypertensive diabetics proved therapeutic since their effect on hypertensive diabetic rats is positive. Therefore, the composite biscuit will offer nutraceutical benefits to both healthy and disease individuals.

Association of placental fatty acid desaturase 2 (FADS2) methylation with maternal fatty acid levels in women with preeclampsia

IntroductionBiosynthesis of long-chain polyunsaturated fatty acids requires sequential activities of desaturases and elongases for conversion of fatty acid precursors to products. The delta-6 desaturase enzyme, encoded by FADS2 gene, is a rate limiting enzyme in this pathway. Alterations in D6D enzyme activity can lead to altered fatty acid profiles. ObjectivesTo examine differences in placental DNA methylation (DNAm) and expression of FADS2 gene in preeclampsia women compared to normal women and their association with maternal variables (plasma fatty acids, desaturase enzyme index, blood pressure), placental weight and birth outcomes. MethodsDNAm and expression of FADS2 gene were examined in placentae of normotensive (n = 100) control and preeclampsia (n = 100) women using pyrosequencing and quantitative real-time PCR respectively. Women with preeclampsia included those delivering at term (n = 43, gestation ≥ 37 weeks; T-PE) or preterm (n = 57, gestation < 37 weeks; PT-PE). A total of 26 CpGs in FADS2 promoter and region around it, were analysed in two PCR reactions (region 1 and 2). ResultsOut of 13 CpGs in region 1, significant hypermethylation was noted at CpG3 in T-PE (p = 0.03) and of 13 CpGs in region 2, CpG2 (p = 0.008), CpG11 (p = 0.04), CpG12 (p = 0.001) were hypomethylated and CpG13 (p = 0.001) was hypermethylated in preeclampsia group, as compared to controls. FADS2 expression was lower in PT-PE as compared to controls (p = 0.04). DNAm at various CpGs in the FADS2 were associated with maternal plasma FADS2 enzyme index and also associated with maternal fatty acid levels. However, we did not observe any association of DNAm with maternal blood pressure, placental weight and birth outcomes. ConclusionsThis study for the first time reports differential methylation of FADS2 and its association with impaired maternal fatty acid metabolism in preeclampsia and provides a mechanistic basis to our earlier observations of altered maternal LCPUFA levels in women with preeclampsia.

Integrated Transcriptome and Metabolomic Analysis Reveal Anti-Angiogenic Properties of Disarib, a Novel Bcl2-Specific Inhibitor.

Transcriptomic profiling of several drugs in cancer cell lines has been utilised to obtain drug-specific signatures and guided combination therapy to combat drug resistance and toxicity. Global metabolomics reflects changes due to altered activity of enzymes, environmental factors, etc. Integrating transcriptomics and metabolomics can provide genotype-phenotype correlation, providing meaningful insights into alterations in gene expression and its outcome to understand differential metabolism and guide therapy. This study uses a multi-omics approach to understand the global gene expression and metabolite changes induced by Disarib, a novel Bcl2-specific inhibitor in the Ehrlich adenocarcinoma (EAC) breast cancer mouse model. RNAseq analysis was performed on EAC mouse tumours treated with Disarib and compared to the controls. The expression of 6 oncogenes and 101 tumour suppressor genes interacting with Bcl2 and Bak were modulated upon Disarib treatment. Cancer hallmark pathways like DNA repair, Cell cycle, angiogenesis, and mitochondrial metabolism were downregulated, and programmed cell death platelet-related pathways were upregulated. Global metabolomic profiling using LC-MS revealed that Oncometabolites like carnitine, oleic acid, glycine, and arginine were elevated in tumour mice compared to normal and were downregulated upon Disarib treatment. Integrated transcriptomic and metabolomic profiles identified arginine metabolism, histidine, and purine metabolism to be altered upon Disarib treatment. Pro-angiogenic metabolites, arginine, palmitic acid, oleic acid, and myristoleic acid were downregulated in Disarib-treated mice. We further validated the effect of Disarib on angiogenesis by qRT-PCR analysis of genes in the VEGF pathway. Disarib treatment led to the downregulation of pro-angiogenic markers. Furthermore, the chorioallantoic membrane assay displayed a reduction in the formation of the number of secondary blood vessels upon Disarib treatment. Disarib reduces tumours by reducing oncometabolite and activating apoptosis and downregulating angiogenesis.

Alterations in Inflammatory Cytokines and Redox Homeostasis in LPS-Induced Pancreatic Beta-Cell Toxicity and Mitochondrial Stress: Protection by Azadirachtin

Inflammation and redox imbalance are hallmarks of cancer, diabetes, and other degenerative disorders. Pathophysiological response to these disorders leads to oxidative stress and mitochondrial dysfunction by alterations and reprogramming in cellular signaling and metabolism. Pancreatic beta cells are very sensitive to the inflammatory and altered nutrient signals and hence play a crucial role in diabetes and cancer. In this study, we treated insulin-secreting pancreatic beta cells, Rin-5F, with the bacterial endotoxin, LPS (1 μg/ml) to induce an inflammatory response in vitro and then treated the cells with a known anti-inflammatory, anticancer and antioxidant phytochemical, azadirachtin (AZD, 25 µM for 24 h). Our results demonstrated lipid peroxidation and nitric oxide production causing increased nitro/oxidative stress and alterations in the activities of anti-oxidant enzymes, superoxide dismutase and catalase after LPS treatment. Pro-inflammatory responses caused by translocation of nuclear factor kappa B and release of inflammatory cytokines were also observed. These changes were accompanied by GSH-dependent redox imbalance and alterations in mitochondrial membrane potential and respiratory complexes enzyme activities leading to mitochondrial respiratory dysfunction, reduced ATP synthesis, and intrinsic caspase-9 mediated apoptosis. Caspase-9 was activated due to alterations in Bcl-2 and Bax proteins and release of cytochrome c into the cytosol. The activities of oxidative stress-sensitive mitochondrial matrix enzymes, aconitase, and glutamate dehydrogenase were also inhibited. Treatment with AZD showed beneficial effects on the recovery of antioxidant enzymes, inflammatory responses, and mitochondrial functions. GSH-dependent redox homeostasis also recovered after the treatment with AZD. This study may help in better understanding the etiology and pathogenesis of inflammation-induced disorders in pancreatic beta cells to better manage therapeutic strategies.

Glycerol monolaurate attenuated immunological stress and intestinal mucosal injury by regulating the gut microbiota and activating AMPK/Nrf2 signaling pathway in lipopolysaccharide-challenged broilers.

This study was conducted to investigate the effects of glycerol monolaurate (GML) on lipopolysaccharide (LPS)-induced immunological stress and intestinal mucosal injury in broilers and its underlying mechanisms. A total of 144 one-d-old Arbor Acres broilers were allocated to a 2 × 2 factorial arrangement involving dietary treatment (0 or 1,200 mg/kg dietary GML) and LPS challenge (injected with saline or Escherichia coli LPS on d 16, 18, and 20). Samples were collected on d 21. The results revealed that dietary GML augmented serum immunoglobulin A (P = 0.009) and immunoglobulin G (P < 0.001) levels in challenged birds. Dietary GML normalized LPS-induced variations in serum interleukin-6, interferon-gamma, and LPS levels (P < 0.05), jejunal villus height (P = 0.030), and gene expression of interleukin-6, macrophage inflammatory protein-3 alpha, Toll-like receptor 4, nuclear factor kappa-B, caspase-1, tight junction proteins, adenosine monophosphate-activated protein kinase alpha 1 (AMPKα1), nuclear factor-erythroid 2-related factor 2 (Nrf2), and superoxide dismutase-1 (P < 0.05). GML supplementation ameliorated LPS-induced peroxidation by reducing malondialdehyde content and increasing antioxidant enzyme activity (P < 0.05). Dietary GML enhanced the abundances of Anaerostipes, Pseudoflavonifractor, and Gordonibacter and reduced the proportion of Phascolarctobacterium in challenged birds. Dietary GML was positively correlated with alterations in antioxidant enzyme activities and AMPKα1, Nrf2, and zonula occludens-1 expressions. The genera Anaerostipes, Lachnospira, Gordonibacter, Lachnospira, Marvinbryantia, Peptococcus, and Pseudoflavonifractor were linked to attenuated inflammation and improved antioxidant capacity of challenged birds. In conclusion, dietary GML alleviated LPS-induced immunological stress and intestinal injury of broilers by suppressing inflammation and oxidative stress. Dietary GML regulated cecal microbiota and activated the AMPK/Nrf2 pathway in LPS-challenged broilers.

Health Benefits of Cereal Grain- and Pulse-Derived Proteins.

Pulses and whole grains are considered staple foods that provide a significant amount of calories, fibre and protein, making them key food sources in a nutritionally balanced diet. Additionally, pulses and whole grains contain many bioactive compounds such as dietary fibre, resistant starch, phenolic compounds and mono- and polyunsaturated fatty acids that are known to combat chronic disease. Notably, recent research has demonstrated that protein derived from pulse and whole grain sources contains bioactive peptides that also possess disease-fighting properties. Mechanisms of action include inhibition or alteration of enzyme activities, vasodilatation, modulation of lipid metabolism and gut microbiome and oxidative stress reduction. Consumer demand for plant-based proteins has skyrocketed primarily based on the perceived health benefits and lower carbon footprint of consuming foods from plant sources versus animal. Therefore, more research should be invested in discovering the health-promoting effects that pulse and whole grain proteins have to offer.

Mfsd8 Modulates Growth and the Early Stages of Multicellular Development in Dictyostelium discoideum.

MFSD8 is a transmembrane protein that has been reported to transport chloride ions across the lysosomal membrane. Mutations in MFSD8 are associated with a subtype of Batten disease called CLN7 disease. Batten disease encompasses a family of 13 inherited neurodegenerative lysosomal storage diseases collectively referred to as the neuronal ceroid lipofuscinoses (NCLs). Previous work identified an ortholog of human MFSD8 in the social amoeba D. discoideum (gene: mfsd8, protein: Mfsd8), reported its localization to endocytic compartments, and demonstrated its involvement in protein secretion. In this study, we further characterized the effects of mfsd8 loss during D. discoideum growth and early stages of multicellular development. During growth, mfsd8 − cells displayed increased rates of proliferation, pinocytosis, and expansion on bacterial lawns. Loss of mfsd8 also increased cell size, inhibited cytokinesis, affected the intracellular and extracellular levels of the quorum-sensing protein autocrine proliferation repressor A, and altered lysosomal enzyme activity. During the early stages of development, loss of mfsd8 delayed aggregation, which we determined was at least partly due to impaired cell-substrate adhesion, defects in protein secretion, and alterations in lysosomal enzyme activity. Overall, these results show that Mfsd8 plays an important role in modulating a variety of processes during the growth and early development of D. discoideum.

Synthesis, Biological Activity and Molecular Docking Studies of Novel Sulfonate Derivatives Bearing Salicylaldehyde.

Enzyme activity alterations have been associated with many metabolism disorders and have crucial roles in the pathogenesis of some diseases. Tyrosinase is a key enzyme in melanin biosynthesis, which is responsible for skin pigmentation to protect the skin from solar radiation. Pancreatic lipase has been considered a key enzyme for the treatment of obesity. Herein, we reported the synthesis and enzyme inhibitions of a series of sulfonates as possible tyrosinase and pancreatic lipase inhibitors. According to the calculated IC50 values, compound 3f (74.1±11.1 μM) and compound 3c (86.6±6.9 μM) were determined to be the best inhibitors among the synthesized compounds for the tyrosinase and pancreatic lipase enzymes, respectively. The approach yielded at extremely high level by creating very flexible structural domains for the chemically modified groups. The structural characterization of the target molecules was implemented by 1 H-NMR, 13 C-NMR, and HR-MS analyses. Also, molecular docking studies of the synthesized compounds with tyrosinase and pancreatic lipase enzymes were conducted using AutoDock Vina software. Additionally, the studies of the absorption distribution, metabolism, and excretion (ADME) were performed to uncover the target compounds' pharmacokinetics, drug similarities, and medicinal properties of the novel sulfonate derivatives bearing salicylaldehyde.

Assessment of Lipid Profile and Oxidative Stress Markers of Persea americana Treated Wistar Rats

The purpose of this study is to assess the effect of ethanolic stem bark extract of Persea americana on lipid profile and oxidative stress markers of male Wistar rats. The rats were randomly divided into three groups of five rats each. They were treated with ethanolic extract of Persea americana (except the control group) for a period of 21days. Group1 received distilled water. Group2 received 200mg/kg of the extract and Group3 received 400mg/kg of the extract. The administration was done for 21days. At the end, the rats were sacrificed and blood sample was collected and sent to the laboratory for analysis. The data obtained was statistically analysed using SPSS software version 21. The result showed that, there was no significant difference in serum total cholesterol, triglyceride, high density lipoprotein and low density lipoprotein when test groups were compared to control. There was a significant decrease in serum very low density lipoprotein level in Group3 (400mg/kg) when compared to Group1 (Control). There was no significant alteration in glutathione reductase, catalase and superoxide dismutase enzyme activities. There was a significant difference in malondialdehyde level (a decrease). In conclusion, the ethanolic extract of Persea americana stem bark exhibited slight hypolipidemic effect because of the significant reduction in Very Low Density Lipoprotein. Also, the extract showed a significant decrease in Malondialdehyde at low dose, which shows that Persea americana may have an anti-oxidative property.