Γ-glutamyltransferase Activity Research Articles

The role of Bacillus strains and growth medium in shaping γ-glutamyl peptide production

To compare γ-glutamyl di- and tripeptides generated by different Bacillus strains and investigate the impact of substrates on the production of γ-glutamyl peptides, six strains of four species, including B. subtilis, B. velezensis, B. amyloliquefaciens and B. paralicheniformis, were cultivated in a standard brain heart infusion (BHI) broth and a medium consisting of hemoglobin hydrolysates (HH) for six days. Quantitative analysis of free amino acids and γ-glutamyl peptides, as well as determination of bacterial growth and γ-glutamyltransferase activity were carried out. Results revealed that all Bacillus strains could generate a series of γ-glutamyl dipeptides in both media. Most strains produced higher concentrations of target peptides in the HH medium (up to 83.56 μM), which correlated with higher levels of free amino acids in the medium. Glutathione was detected only in the BHI medium with B. subtilis PRO84, B. velezensis PRO76, B. altitudinis PRO107, and B. paralicheniformis PRO109 (up to 0.61 μM), indicating the glutathione-forming ability of these Bacillus strains. Production of γ-glutamyl peptides was influenced by both the choice of strain and growth medium, with the medium exerting a more pronounced impact than the strain. This study highlights the importance of selecting the appropriate substrate and Bacillus strains to produce desired γ-glutamyl peptides as enhancers of koku, potentially inspiring the conversion of meat side streams into valuable kokumi seasonings.

Preconditioning by Moderate-Intensity Exercise Prevents Gentamicin-Induced Acute Kidney Injury.

A strict correlation among proximal tubule epithelial cell dysfunction, proteinuria, and modulation of the Renin-Angiotensin System and Kalikrein-Kinin System are crucial factors in the pathogenesis of Acute Kidney Injury (AKI). In this study, we investigated the potential protective effect of preconditioning by moderate-intensity aerobic exercise on gentamicin-induced AKI. Male Wistar rats were submitted to a moderate-intensity treadmill exercise protocol for 8 weeks, and then injected with 80 mg/kg/day s.c. gentamicin for 5 consecutive days. Four groups were generated: 1) NT+SAL (control); 2) NT+AKI (non-trained with AKI); 3) T+SAL (trained); and 4) T+AKI (trained with AKI). The NT+AKI group presented: 1) impairment in glomerular function parameters; 2) increased fractional excretion of Na + , K + , and water; 4) proteinuria and increased urinary γ-glutamyl transferase activity (a marker of tubular injury) accompanied by acute tubular necrosis; 5) an increased renal angiotensin-converting enzyme and bradykinin B1 receptor mRNA expression. Interestingly, the preconditioning by moderate-intensity aerobic exercise attenuated all alterations observed in gentamicin-induced AKI (T+AKI group). Taken together, our results show that the preconditioning by moderate-intensity aerobic exercise ameliorates the development of gentamicin-induced AKI. Our findings help to expand the current knowledge regarding the effect of physical exercise on kidneys during physiological and pathological conditions.

Inositol-6-phosphate mitigates deoxynivalenol nephrotoxicity in pig kidney explants

Abstract Deoxynivalenol (DON) is a mycotoxin that frequently contaminates food crops and negatively impacts human and animal health. While inositol-6-phosphate (IP6) is known to counteract DON-induced intestinal damage, inflammation, and oxidative stress, its ability to alleviate DON-caused kidney injury is yet unexplored. This study assessed DON’s kidney effects and IP6’s protective capability using an ex vivo renal explants model. From 360 explants taken from six pigs, four groups were formed: control (culture medium), DON (10 μM equivalent to 3 mg/kg of feed), IP6 group (5 mM), and DON + IP6 (10 μM + 5 mM). DON elevated creatinine levels and γ-glutamyl transferase activity in the culture medium. It led to cytoplasmic vacuolation in convoluted tubular epithelial cells and tubular necrosis, a drop in antioxidant potential (ABTS) and a rise in interleukin (IL)-8 mRNA. The introduction of IP6 restored renal histological and functional parameters, bolstered antioxidant status, enhanced IL-10 gene expression, and reduced superoxide anion levels. These results indicate that within the model of renal explants, IP6 significantly alleviates the nephrotoxic effects of DON while efficiently regulating the antioxidative and anti-inflammatory response within the kidneys.

Exploring the anticancer potential of double-active pharmaceutical ingredient, metforminium ibuprofenate: Integrating in vivo and in silico investigations

Rationale: A novel double active pharmaceutical ingredient (d-API), metforminium ibuprofenate (MtHIb) was synthesised by combining metformin hydrochloride (MtHCl) and sodium ibuprofen (NaIb). This formulation exhibits anti-diabetic and anti-inflammatory properties similar to its parent compounds, along with a remarkable antiproliferative effect on diverse cancer cells. AimThe present study is aimed to analyse the anticancer potential of MtHIb by mouse tumour model and molecular docking. MethodsSolid tumour was induced by Dalton's Lymphoma Ascites (DLA) cells in Swiss albino mice, and d-API was administered orally. The tumour volume was measured using Vernier calliper and the activity of γ-glutamyl transferase (GGT) was assessed. The potential interactions of MtHIb with target proteins mTOR, HER2, EGFR, CDK4, CDK6, ERα/β, progesterone, COX2, BUB1, and KEAP2 were analyzed in silico. ResultsA significant reduction in tumour size was observed in mice treated with MtHIb. On the 30th day, the tumour volume, which was 8.12 cm3 in untreated animals, decreased to 1.311 cm3 with the treatment of 5 mg/kg b.wt of MtHIb, representing an 83.8 % reduction. The activity of GGT, a marker enzyme of proliferative activity, was considerably reduced from 394.87 to 51.26 U/mg protein/L. Molecular docking studies indicated that MtHIb exhibited strong binding affinity towards all the screened target proteins, specifically HER2, EGFR, CDK4, and BUB1. DiscussionThe findings suggest that MtHIb has promising anti-tumour effect, as evidenced by its significant reduction in tumour size and GGT activity in mice. The strong binding affinity of MtHIb towards the key cancer related proteins further supports its potential as a therapeutic agent against cancer, particularly in targeting pathways involved in cell proliferation and survival. Further preclinical and clinical studies are required to validate these findings and to explore the full therapeutic potential of MtHIb in cancer treatment.

Effects of graded levels of dietary pomegranate peel on methane and nitrogen losses, and metabolic and health indicators in dairy cows

This study aimed to quantify the effects of dietary inclusion of tannin-rich pomegranate peel (PP) on intake, methane and nitrogen (N) losses, and metabolic and health indicators in dairy cows. Four multiparous, late-lactating Brown Swiss dairy cows (796 kg body weight; 29 kg/d of energy corrected milk yield) were randomly allocated to 3 treatments in a randomized cyclic change-over design with 3 periods, each comprising 14 d of adaptation, 7 d of milk, urine, and feces collection, and 2 d of methane measurements. Treatments were formulated using PP that replaced on a dry matter (DM) basis 0% (control), 5%, and 10% of the basal mixed ration (BMR) consisting of corn and grass silage, alfalfa, and concentrate. Gaseous exchange of the cows was determined in open-circuit respiration chambers. Blood samples were collected on d 15 of each period. Individual feed intake as well as feces and urine excretion were quantified, and representative samples were collected for analyses of nutrients and phenol composition. Milk was analyzed for concentrations of fat, protein, lactose, milk urea N, and fatty acids. Total phenols and antioxidant capacity in milk and plasma were determined. In serum, the concentrations of urea and bilirubin as well as the activities of alanine aminotransferase (ALT), aspartate aminotransferase, glutamate dehydrogenase, alkaline phosphatase, and γ-glutamyl transferase were measured. The data were subjected to ANOVA with the Mixed procedure of SAS, with treatment and period as fixed and animal as random effects. The PP and BMR contained 218 and 3.5 g of total extractable tannins per kg DM, respectively, and thereof 203 and 3.3 g of hydrolyzable tannins. Total DM intake, energy corrected milk, and methane emission (total, yield, and intensity) were not affected by PP supplementation. The proportions of C18:2n-6 and C18:3n-3 in milk increased linearly as the amount of PP was increased in the diet. Milk urea N, blood urea N, and urinary N excretion decreased linearly with the increase in dietary PP content. Total phenols and antioxidant capacity in milk and plasma were not affected by the inclusion of PP. The activity of ALT increased in a linear manner with the inclusion of PP. In conclusion, replacing up to 10% of BMR with PP improved milk fatty acid composition and alleviated metabolic and environmental N load. However, the elevated serum ALT activity indicates an onset of liver stress even at 5% PP, requiring the development of adaptation protocols for safe inclusion of PP in ruminant diets.

The effect of mineral water on the course of experimental oxalate nephrolithiasis

Introduction. The number of patients with urolithiasis (UCD) is increasing annually both among the adult population and among children, and therefore it is necessary to develop an integrated approach to the prevention and treatment of this pathology. Most often, with urolithiasis, oxalate-calcium crystalluria is determined, to reduce the level of which, low-mineralized mineral waters with a minimum sodium content are recommended.Aim. To study the effect of mineral water “Serebryanyy klyuch” on the course of experimental calcium oxalate nephrolithiasis.Materials and methods. During the study on laboratory rats, oxalate nephrolithiasis was simulated for 3 weeks by providing a 1% solution of ethylene glycol as a drink, while the rats of the experimental group used mineral water “Serebryanyy klyuch” as a solvent. The results of urine biochemistry (calcium, creatinine, activity of lactate dehydrogenase and γ-glutamyl transferase) and morphological studies (determination of the amount and size of calcium deposits in the kidneys, assessment of the structural and functional state of the nephrothelium) were evaluated.Results. Calcium-oxalate deposits in the control group of rats were detected in 25% of cases, while large-sized stones were observed in the kidney tubules. In the experimental group, the presence of stone deposits was detected in 1 (11.1%) case, with a clear shift towards smaller deposits. Morphological signs indicating the development of dystrophic changes with the phenomena of nephrosclerosis were much less pronounced in the conditions of consumption of mineral water “Serebryanyy klyuch”.Conclusion. Thus, the use of mineral water “Serebryanyy klyuch” in the conditions of a three-week experimental oxalate nephrolithiasis was accompanied by characteristic signs of facilitating the course of the pathology and contributed to a decrease in the proportion of large calculi in the kidneys.

Assessing the molluscicidal and biochemical activities of New-Fort®, an inorganic fertilizer against Theba pisana snails

Terrestrial snails are a significant issue in agricultural production worldwide. The use of nitrogen - phosphorus - potassium (NPK) based fertilizers played an important role in meeting the food demand throughout the world, so its effectiveness against land snails needs to be investigated. This study was conducted to evaluate toxic lethal effect of New-Fort®, an inorganic NPK based fertilizer, in the field for 3, 7 and 10 days and in the laboratory for 24, 48 and 72 h against Theba pisana snails. Also, the impact of its sub-lethal doses (1/10, 1/5, 1/4 and 1/2 of 48 h-LD50) on biochemical parameters were determined under laboratory conditions. The results showed that the snails percent reduction in the field were 21.4, 61.0 and 80.0 % after 10 days' application of quarter, half and one field rate and the values of LD50 in the laboratory were 4.94, 4.56 and 4.24 mg/g b.w at 24, 48 and 72 h, respectively. New-Fort® sub-lethal doses caused a significant inhibition in catalase, γ-glutamyl transferase and acetylcholinesterase activities. It also elicited a significant elevation in glutathione S-transferase activity post exposure to 1/10 and 1/5 of LD50, whereas an opposite effect was occurred after exposure to 1/4 and 1/2 of LD50. Lipid peroxidation level was reduced in snails treated with 1/10 and 1/5 of LD50, whereas it increased in 1/4 and 1/2 of LD50- treated snails. Moreover, a significant inhibition in alkaline phosphatase activity at all tested doses, with the exception of 1/2 of LD50 was observed. An increase in alanine aminotransferase and aspartate aminotransferase activities were occurred after all tested doses exposure. Our findings highlighted on how biochemical changes can be exploited to better understand the mechanisms underlying New-Fort® fertilizer toxicity against the land snail, T. pisana, as well as how to benefit from NPK fertilizers application in snail control.

Bio-molluscicidal potential and biochemical mechanisms of clove oil and its main component eugenol against the land snail, Theba pisana

The land snail, Theba pisana is a serious pest that adversely affects various crops in sustainable agriculture. Essential oils and their constituents represent an environmentally sound alternative to synthetic pesticides. Our study aimed to investigate the lethal and sub-lethal toxicity of clove oil and its main component eugenol to understand the mechanisms underlying its toxic action against T. pisana. The GC–MS profile of the clove oil composition was characterized. In the laboratory experiment, LD50 of clove oil and eugenol via the contact testing were determined after 48 and 72 h. Moreover, sub-lethal effects of clove oil or eugenol on the survivors following the exposure of snails to the 25 and 50% of the LD50/48 and 72 h were evaluated through using snail tissues for biochemical measurments. The GC–MS analysis showed that eugenol (64.87%) was the major constituent present in the oil. The results also showed that LD50 values at 48 and 72 h were 2006.5 and 1493.5 μg/g b.w for oil and 239.6 and 195.3 μg/g b.w for eugenol, respectively. Compared to control, the sub-lethal effects of clove oil or eugenol at 48 and 72 h showed a significant increase in reduced glutathione (GSH) levels. Catalase (CAT) and glutathione-S-transferase (GST) activities significantly elevated in oil- or eugenol-treated snails, except at low dose after 48 h. After two exposure times, snails exposed to oil or eugenol at both sub-lethal effects had considerably higher γ-glutamyltransferase (γ-GT) and aspartate aminotransferase (AST) activities. Moreover, markedly augmentation in alkaline phosphatase (ALP) and alanine aminotransferase (ALT) activities at all exposure times, with the exception of snails treated with low dose of eugenol after 48 h was observed. Both clove oil and eugenol at the tested doses caused a significant inhibition in acetylcholinesterase (AChE) activity at two exposure times. Our findings highlight the potential of clove oil and eugenol, as an efficient natural molluscicide alternative to its synthetic counterparts for snail control.

Neonatal sclerosing cholangitis with novel mutations in DCDC2 (doublecortin domain-containing protein 2) in Chinese children.

Neonatal sclerosing cholangitis (NSC) is a rare and severe autosomal recessive inherited liver disease with mutations in DCDC2, commonly requiring liver transplantation (LT) for decompensated biliary cirrhosis in childhood. The information of four Chinese patients with NSC caused by mutations in DCDC2 from Children's Hospital of Fudan University were gathered. The four patients' clinicopathological and molecular features were summarized by clinical data, liver biopsy, immunohistochemical, and molecular genetic analysis. All patients presented with jaundice, hepatosplenomegaly, hyperbilirubinemia and bile embolism, and high serum γ-glutamyl transferase activity (GGT). Liver biopsies revealed varying degrees of bile duct hyperplasia, portal-tract inflammation, and/or fibrosis. Whole-exome sequencing (WES) found novel heterozygous variants of c.1024-1G > T /p.? and c.544G > A /p. Gly182Arg in the DCDC2. This study expands the genetic spectrum of DCDC2 in NSC.

In vivo exposure to pyriproxyfen causes ovarian oxidative stress and inhibits follicle maturation in zebrafish.

We analyzed the effects of pyriproxyfen (PPF) on oxidative stress and ovarian morphology in zebrafish. PPF (10-9 M) exposure increased reactive oxygen species generation in ovaries, in association with a decrease in glutathione content. The activities of glutathione S-transferase, superoxide dismutase, and catalase were increased, while γ-glutamyltransferase activity was not altered by pesticide treatment. The histology of ovarian tissue showed an increase in the number of previtellogenic oocytes I, and a decrease in the rate of vitellogenic oocyte (VIT) count, suggesting inhibition of follicular maturation. An increase in the thickness of the vitelline envelope was observed in VIT, as was a tendency toward an increase in atresia in the ovary of the PPF-treated group. These findings indicate that the deleterious effect of PPF on ovarian maturation is mediated by a redox imbalance and oxidative damage. So, PPF acts as an endocrine disruptor chemical and may compromise fish reproduction by reducing female fertility.

Effects of triterpenoids in fatty products on liver condition of laboratory animals with acute toxic hepatitis

One of the principles of prevention and non-medicamentous treatment of liver diseases, including hepatitis of different etiology, is the normalization of the diet through the consumption of food with physiologically active ingredients, in particular betulin, which helps to eliminate the causes of metabolic and oxidative disorders within liver cells. The aim of the research was to assess in vivo the influence of triterpene alcohol betulin extracted from Betula pendula Roth. birch bark in fat-containing products (for example mayonnaise) on the blood biochemical parameters and liver morphological structure of rats with initiated acute toxic hepatitis. Material and methods. Hepatoprotective and antioxidant activities of betulin as part of mayonnaise samples has been investigated in vivo on the model of toxic hepatitis initiated by carbon tetrachloride in male Wistar rats weighing 210-265 g. The animals were divided into 4 groups of 10 animals each: CG-1 - intact, CG-2 and MG - with carbon tetrachloride initiated toxic hepatitis. rats of the main groups were orally administered mayonnaise once a day at a dosage of 1 ml for 21 days after the formation of the model pathology: OG-1 with the added betulin (1 mg per 1 kg of body weight), OG-2 without betulin. Disorders of metabolic and oxidative processes in liver cells of animals were evaluated by biochemical indicators of blood plasma: the level of glucose, albumin, total cholesterol, triglycerides and urea and the activity of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and γ-glutamyltransferase. Oxidative stress in rats was estimated by the activity of catalase and superoxide dismutase in blood hemolysate (at a dilution of 1:200 and 1:10, respectively); the total prooxidant (in blood plasma) and total antioxidant (in blood hemolysate at a dilution of 1:10) activity were determined spectrophotometrically (colored complexes of TWIN-80 oxidation products with thiobarbituric acid). The morphological structure of rats' liver was estimated by microscopy of prepared cuts of hepatic tissue. Results. Based on biochemical parameters of rat blood plasma, it has been established that the administration of mayonnaise with betulin prevents the development of cytolic syndrome and suppresses the process of peroxidation by directly neutralizing free radicals. Aspartate aminotransferase and alkaline phosphatase activity in blood plasma of the experimental animals of the main group MG-1 reduced by 20.7 and 35.2% compared with indicators of the rats of the main group MG-2. Glucose concentration normalized to the level of the control group CG-1. The concentration of bilirubin and triglycerides decreased by 22.9 and by 48.1%, which indicates a significant reduction in the indicators of cholestatic syndrome in the group of animals OG-1 compared to OG-2. The total prooxidant activity and the concentration of thiobarbiturate-reactive products decreased compared to the CG-2 and MG-2 groups, which indicates the suppression of oxidative stress and, as a result, an improvement in liver conditions of animals with toxic hepatitis even when taking a fat-containing product. In liver histopeparates of animals receiving mayonnaise with betulin, necrobotic changes were less pronounced in comparison with the group MG-2. They were estimated at 1 point: small-drip dystrophy spots were found, haemorrhages in the interregional septum with inflammatory infiltration in the course of hemorrhages against the presence of necrosis hepatocytes with pronounced adipose dystrophy in the centres of the lobules, step necrosis with signs of replacing the damaged hepatocytes of the connective tissue, accompanied by centrolobular hemorrhages in MG-2 rats. Conclusion. Introduced into the composition of mayonnaise betulin, reduces the development of cytolic syndrome in toxic hepatitis and suppresses the process of peroxidation, on the basis of which fat-containing foods with betulin can be recommended for clinical examination as specialized products in acute and chronic liver diseases, including complicated cholestasis.

Effects of crude protein and neutral detergent fiber percentages in the diet of Japanese Black steers on rumen fluid properties, blood biochemical properties, and carcass characteristics.

The effects of crude protein (CP) and neutral detergent fiber (NDF) percentages in the diet of Japanese Black steers on rumen fluid properties, blood biochemical properties, and carcass characteristics were examined. Twelve 13-month-old Japanese Black steers were used for this study and slaughtered at 30 months of age. Steers were assigned to a control group (n = 6) and test group (n = 6) and were fed a concentrate containing 12.9%-13.9% CP and 26.5%-29.8% NDF or 9.1%-9.6% CP and 29.9%-31.2% NDF, respectively. Lipopolysaccharide activity levels in rumen fluid were lower in the test group than in the control group. Plasma urea nitrogen concentration and activities of aspartate aminotransferase and γ-glutamyltransferase remained lower in the test group than in the control group. In contrast, plasma vitamin A concentrations remained higher in the test group than in the control group. Carcass characteristics did not significantly differ between the two groups. These results suggest that dietary CP and NDF percentages in feed for Japanese Black steers older than 13 months of age affected rumen fluid properties and blood biochemical properties, indicating a reduced load on the liver with a small effect on carcass characteristics.

Transient increases in glutamate dehydrogenase (GLDH) activity occur in neonatal foals.

Liver analyte measurement is important in the evaluation of sick animals. Liver injury in horses is recognized by increased glutamate dehydrogenase (GLDH), sorbitol dehydrogenase (SDH), and aspartate aminotransferase (AST) activities, whereas biliary pathology is identified by increased alkaline phosphatase and γ-glutamyl transferase (GGT) activities or bilirubin concentrations. We have observed high GLDH, but not SDH, activities in neonatal foals admitted for conditions other than liver disease. Only one previous study have evaluated GLDH activity over time in healthy neonatal foals; however, SDH activity was not measured. We aimed to evaluate changes in liver analytes in neonatal foals over time. We measured serum liver analytes (GLDH, SDH, GGT, AST, total, direct, and indirect bilirubin) and creatine kinase activity of 11 clinically healthy foals before and at various times after suckling until 46 days of age. Analytes were also measured in colostrum and mare serum. Median GLDH activities increased after birth to peak at 3-4 days of age (106 U/L, reference interval, 0-8U/L). Median SDH activities had a lower peak at 3-4 days (15 U/L, reference interval, 0-11 U/L) and were frequently discordant with GLDH. There was no association between foal and mare serum or colostral enzyme activities. AST activity plateaued at 5-6 days, whereas GGT activity and total and indirect bilirubin concentrations peaked at 14 and 3-4 days of age, respectively. Transient increases in GLDH, SDH, and GGT activities and total and indirect bilirubin concentrations occur in clinically healthy neonatal foals and do not necessarily indicate relevant liver disease.

Norcantharidin toxicity profile: an in vivo murine study.

Norcantharidin (NCTD) is the demethylated analog of cantharidin, with allegedly reduced toxicity. However, there is still limited information regarding its posology and potential risk in its use in cancer treatment. Healthy BDF1 mice were intraperitoneally administered with norcantharidin (0, 3, 6, 12, and 25mg/kg) every 24h for 6days. Survivor mice were euthanized, and the brain, lungs, kidneys, spleen, and liver were procured for enzymatic and histopathological analysis in the liver and kidney. DL50 were 8.86mg/kg for females and 11.77mg/kg for males. The treatments with 3.0mg/kg and 6.0mg/kg significantly modified the phosphorylase, alanine transaminase, and γ-glutamyl transferase activities; however, an organ-specific response was detected. A significant dose-dependent decrease was observed in the kidney for ROS, while the liver had the opposite effect. Histopathological analysis revealed a significant elevation in hepatocytes' nuclei average size and total area (3mg/kg), as well as centrilobular vein and adjacent sinusoidal capillaries showed a significant difference. The portal triad presented a significant difference in veins and capillarity count in 6mg/kg. Renal samples showed cortex convoluted tubules' average size significantly augmented in both doses' groups, and tubule count was found augmented in 6mg/kg. These physiological effects of NCTD can be exploited as treatment strategies if able to operate in an established posology and proper testing.

A water-soluble fluorescent probe for the determination of γ-glutamyltransferase activity and its application in tumor imaging

γ-glutamyltransferase (GGT), an important tumor marker, is highly expressed in tumor tissues, and precise detection of its activity provides a vital indicator for the diagnosis and treatment. In this work, a “lighting-on” probe (TCF-GGT) was elaborated to detect endogenous GGT with high selectivity and sensitivity. Dicyanomethyldifuranyl (TCF-OH) was employed as the fluorescence reporter and short peptide glutathione (GSH) worked as the GGT-active trigger, the introduction of which prevented the initial proton transfer of TCF-OH contributing to a blank sensing background. A bright red fluorescence could be switched on upon GGT catalytic hydrolysis, avoiding the potential interference from background. There displayed an excellent water-solubility, and little organic solvent was required during the exploration, which otherwise avoided the potential damage to enzyme and organism. TCF-GGT has been proved to be workable at cellular and organism level with highly effective imaging and a short metabolic cycle, which is expected to offer an alternative solution or reference to the early diagnosis and treatment of tumor.

Red Palm Oil Ameliorates Oxidative Challenge and Inflammatory Responses Associated with Lipopolysaccharide-Induced Hepatic Injury by Modulating NF-κβ and Nrf2/GCL/HO-1 Signaling Pathways in Rats.

Lipopolysaccharide (LPS), a well-conserved cell wall component of Gram positive bacteria, exerts its toxic effects via inducing oxidative and pro-inflammatory responses. Red palm oil (RPO) is a unique natural product with a balanced ratio of saturated and unsaturated fatty acids, with reported antioxidant and anti-inflammatory effects. In this study, we assess the protective effect and mechanistic action of RPO using a lipopolysaccharide (LPS)-induced hepatic injury model. Male Wistar rats were assigned into four groups (10 animals/group): normal control (NC), RPO, LPS and RPO + LPS. Animals in the RPO and RPO + LPS groups were administered RPO (200 μL/day) for 28 days. On the 27th day of experiment, animals in LPS and RPO + LPS groups were injected with LPS (0.5 mg/kg body weight). Animals were sacrificed 24 h later, and blood and liver tissues harvested for biochemical and molecular analysis. RPO resolved hepatic histological dysfunction induced by LPS, and lowered alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and γ-glutamyl transferase activities in the serum. Hepatic malondialdehyde and conjugated dienes, as well as pro-inflammatory cytokines, including interleukin (IL)-1β, IL-6 and TNFα were significantly diminished (p < 0.05) by RPO pre-treatment. Activity of hepatic antioxidant enzymes including superoxide dismutase, glutathione reductase, glutathione peroxidase, as well as glutathione redox status (GSH:GSSG), and markers of antioxidant capacity that decreased as a result of LPS injection were improved by RPO pre-treatment. Mechanistically, RPO up-regulated mRNA expression of redox sensitive transcription factor Nrf2 and its downstream targets GCL and HO-1, while also suppressing the expression of NFκβ and associated inflammatory protein, Iκβ kinase (IκKβ). In conclusion, this study highlights the ameliorating effects of RPO against LPS-induced hepatic injury and revealed the Nrf2/GCL/HO-1 and NFκβ signaling axis as potential contributing mechanisms.

Influence of growth medium and yeast species on the formation of γ-glutamyl peptides

γ-Glutamyl peptides are known to impart a kokumi sensation and increase the palatability of food. To compare the γ-glutamyl peptides produced by different yeasts, Saccharomyces cerevisiae and Debaryomyces hansenii were cultured in a conventional medium (YPG) and a medium composed of porcine hemoglobin hydrolysates (YHG) for six days. Quantification of γ-glutamyl peptides as well as analysis of yeast growth and γ-glutamyltransferase (GGT) activity were carried out. Results showed that both strains could form a range of γ-glutamyl dipeptides and glutathione in either medium. The total concentration of dipeptides and the concentration of glutathione in each medium showed an upward trend over fermentation time. Compared with yeast species, medium composition had a greater impact on the formation of γ-glutamyl dipeptides. During most of the fermentation period, D. hansenii produced more γ-glutamyl dipeptides (reaching a maximum of 2.395 μM on day 6 in YHG), while S. cerevisiae generated more glutathione (reaching a maximum of 13.465 μM on day 5 in YHG). In addition, D. hansenii showed higher GGT activity than S. cerevisiae in the latter half of fermentation. This study illustrates the influence of growth medium and the crucial choice of yeast species for obtaining target γ-glutamyl peptides for kokumi sensation.

Protective effect of protexin concentrate in reducing the toxicity of chlorpyrifos in common carp (Cyprinus carpio)

The present study aimed to evaluate the protective effect of protexin supplementation against chlorpyrifos-induced oxidative stress and immunotoxicity in Cyprinus carpio. After 21 days, the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GR), and total antioxidant levels significantly decreased in hepatocytes of fish exposed to chlorpyrifos, while malondialdehyde (MDA) increased. Treatment with protexin was able to reverse the decrease in SOD and GR and significantly reduce MDA levels. Exposure to chlorpyrifos also induced alterations in blood biochemical parameters and caused immunosuppression. Dietary protexin return some parameters (aspartate aminotransferase, lactate dehydrogenase, and γ-glutamyltransferase activities, and glucose, cholesterol, total protein, creatinine, and complement C4 levels) to values similar to those of the control group. Based on the results, it can be concluded that protexin exerted protective effects against chlorpyrifos exposure in C. carpio reducing oxidative damage, and ameriorating blood biochemical alterations and the immunosuppression.

Assessment of urine sample quality by the simultaneous measurement of urinary γ-glutamyltransferase and lactate dehydrogenase enzyme activities: possible application to unravel cheating in drugs of abuse testing.

Evaluation of the simultaneous measurement of urinary γ-glutamyltransferase (γGT) and lactate dehydrogenase (LDH) to discriminate fresh from previously frozen specimens in urine drug monitoring. Two widely available photometric tests (Siemens Healthineers Atellica) were used to determine the range of urinary γGT and LDH excretion and to study the decay in urinary enzyme activity under various storage conditions (room temperature, 4-8°C,-18°C,-80°C). From these data, cut-off values were established and evaluated in split (fresh/frozen) specimens. Both assays allow robust, reliable, and simultaneous determination of urinary γGT and LDH. In healthy subjects, the 95% reference intervals for enzyme activity in native urine were γGT: 24.4-100.4U/g Crea (creatinine) and LDH: 2.5-45.8U/g Crea. Frozen storage for at least 7days at-18°C resulted in a loss of activity to less than 50% in both enzymes. Cut-offs for frozen samples were γGT≤33.2U/g Crea and LDH≤ 8.4U/g Crea. When applied to 100 sample pairs (fresh/frozen), 86.5% (173/200) of the measurements were conclusive and the combination of concordant enzyme measurements (low γGT/low LDH or high γGT/high LDH) was able to predict the mode of storage with a sensitivity of 96.3% and a specificity of 96.7%. The additional measurements of urinary γGT and LDH can be used to detect previously frozen urine specimens. A simple protocol is proposed to provide additional information on sample quality when deceit is suspected. The procedure can be easily integrated into the standard workflow of urinary drug monitoring.

Extruded Enzyme-Added Corn Improves the Growth Performance, Intestinal Function, and Microbiome of Weaning Piglets.

Simple SummaryEnergy feeds such as corn, wheat, and barley usually contain large amounts of raw starch, which is generally required to be extruded for animals. In addition, amylase preparations are widely added into the diet to degrade raw starch and promote its digestion in livestock animals. The in vitro degradation of corn starch could be a strategy for improving the starch digestion, as it is unknown whether the starch structure and its in vivo digestion could be improved by adding amylase during extrusion, regulating the extrusion moisture content. In this study, we aimed to investigate the effects of extruded corn with added amylase under different moisture conditions on the growth performance, intestinal function, and microbiome of weaning piglets.The objective of this study was to evaluate the effects of extruded corn with added amylase under different moisture conditions on the growth performance, intestinal function, and microbiome of weaning piglets. Fourty-eight 24-day-old weaning piglets (Duroc × Landrace × Yorkshire, weaned at 22 ± 1 d) with an initial body weight of 6.76 ± 0.15 kg were randomly assigned to one of four dietary treatments with six replicates per treatment and two pigs per replicate: (1) NL (adding 7.5% water before corn extrusion, negative treatment with low moisture); (2) NH (adding 15.0% water before corn extrusion, negative treatment with high moisture); (3) PL (adding 7.5% water and 4 kg/t α-amylase before corn extrusion, positive treatment with low moisture); and (4) PH (adding 15% water and 4 kg/t α-amylase before corn extrusion, positive treatment with high moisture). Results showed that amylase supplementation (4 vs. 0 kg/t) increased the contents of small molecular oligosaccharides of extruded corn (p < 0.05). Amylase supplementation significantly improved the average daily feed intake, apparent total tract digestibility (ATTD) of dry matter, crude protein, gross energy, crude fat, ash, phosphorus, and calcium, and also increased the activities of jejunal trypsin, α-amylase, lipase, sucrase, maltase, γ-glutamyl transferase and alkaline phosphatase activities, improved the duodenal, jejunal and ileal morphology, and increased the relative mRNA expressions of the ZO-1, OCLN, SGLT1, and GLUT2 genes in the jejunum (p < 0.05), whereas it decreased the contents of isobutyric acid in cecal digesta, as well as acetic acid and isobutyric acid in colonic digesta (p < 0.05). Moreover, the linear discriminant analysis effect size (LEfSe) showed that piglets fed extruded corn with added enzymes contained less intestinal pathogenic bacteria, such as Holdemanella and Desulfovibrio, compared with piglets fed just extruded corn. In summary, the results of the present study indicated that the supplementation of α-amylase during the conditioning and extruding process of corn increased the small molecular oligosaccharide content of corn starch. Moreover, piglets receiving extruded enzyme-added corn had better growth performance, which was associated with the improved intestinal digestive and absorptive function, as well as the intestinal microbiome.