Γ-glutamyl Transpeptidase Activity Research Articles

IMPACT OF CORONARY ARTERY DISEASE ON STEATOTIC LIVER DISEASE PROGRESSION

Introduction. Metabolic dysfunction-associated steatotic liver disease and coronary artery disease often exacerbate each other, leading to a more severe course of both conditions. The aim of this study is to evaluate the role of coronary artery disease in dislipidemia and liver fibrosis progression in patients with metabolic dysfunction-associated steatotic liver disease. Materials and methods. Forty-six patients with metabolic dysfunction-associated steatotic liver disease were examined. Patients were divided into two groups based on the presence of concomitant coronary artery disease: group I (n=24) included patients with both metabolic dysfunction-associated steatotic liver disease and coronary artery disease, while group II (n=22) included patients with metabolic dysfunction-associated steatotic liver disease only. A third group (Group III) comprised healthy individuals. Specific lipid metabolism parameters, obesity stage, alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGT) activity, and platelet count were examined. Fibrosis stage was evaluated using the FIB-4 score. The findings obtained were statistically processed. Results. Arterial hypertension was more prevalent in patients with metabolic dysfunction-associated steatotic liver disease and concomitant coronary artery disease. In group I, arterial hypertension was observed in 95.8% of cases, whereas in group II, it was observed in 40.9% of cases (χ2=16.35, p=0.00005). Patients with metabolic dysfunction-associated steatotic liver disease exhibited elevated levels of total cholesterol, low-density lipoproteins, and triglycerides, leading to an increased atherogenic index. Conversely, high-density lipoprotein levels were decreased (p>0.05). The degree of fibrosis according to the FIB-4 score was 1.45 times higher in patients with metabolic dysfunction-associated steatotic liver disease and concomitant coronary artery disease compared to those without coronary artery disease.

Synergistic Protective Effect of Fermented Schizandrae Fructus Pomace and Hoveniae Semen cum Fructus Extracts Mixture in the Ethanol-Induced Hepatotoxicity.

Schizandrae Fructus (SF), fruits of Schisandra chinensis (Turcz.) Baill. and Hoveniae Semen cum Fructus (HSCF), the dried peduncle of Hovenia dulcis Thunb., have long been used for alcohol detoxification in the traditional medicine of Korea and China. In the current study, we aimed to evaluate the potential synergistic hepatoprotective effect of a combination mixture (MSH) comprising fermented SF pomace (fSFP) and HSCF hot water extracts at a 1:1 (w:w) ratio against ethanol-induced liver toxicity. Subacute ethanol-mediated hepatotoxicity was induced by the oral administration of ethanol (5 g/kg) in C57BL/6J mice once daily for 14 consecutive days. One hour after each ethanol administration, MSH (50, 100, and 200 mg/kg) was also orally administered daily. MSH administration significantly reduced the serum activities of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and γ-glutamyl transpeptidase. Histological observation indicated that MSH administration synergistically and significantly decreased the fatty changed region of hepatic parenchyma and the formation of lipid droplet in hepatocytes. Moreover, MSH significantly attenuated the hepatic triglyceride accumulation through reducing lipogenesis genes expression and increasing fatty acid oxidation genes expression. In addition, MSH significantly inhibited protein nitrosylation and lipid peroxidation by lowering cytochrome P450 2E1 enzyme activity and restoring the glutathione level, superoxide dismutase and catalase activity in liver. Furthermore, MSH synergistically decreased the mRNA level of tumor necrosis factor-α in the hepatic tissue. These findings indicate that MSH has potential for preventing alcoholic liver disease through inhibiting hepatic steatosis, oxidative stress, and inflammation.

A Mixture of Fermented Schizandrae Fructus Pomace and Hoveniae Semen cum Fructus Extracts Synergistically Protects against Oxidative Stress-Mediated Liver Injury.

Schizandrae Fructus (SF) and Hoveniae Semen cum Fructus (HSCF) have long been used as medicinal herbs for treating various diseases in Asian traditional medicine. In the current study, we investigated the protective effect of fermented SF pomace and HSCF extract 1:1 (w:w) combination mixture (MSH) against carbon tetrachloride (CCl4)-induced acute liver injury mice. After MSH (50-200 mg/kg) oral administration for 7 consecutive days, animals were injected intraperitoneally with CCl4 (0.5 mL/kg). Histopathological observation revealed that administration of MSH synergistically decreased the degeneration of hepatocytes and the infiltration of inflammatory cells induced by CCl4. Moreover, MSH administration reduced the activities of alanine aminotransferase, aspartate aminotransferase, and γ-glutamyl transpeptidase in serum, and mitigated apoptotic cell death in hepatic parenchyma. In addition, MSH alleviated CCl4-mediated lipid peroxidation by restoring endogenous antioxidants capacities including glutathione contents, superoxide dismutase, and catalase activities. In vitro assessments using tert-butyl hydroperoxide-induced oxidative stress in HepG2 cells revealed that MSH protects hepatocytes by lowering ROS generation and lipid peroxidation via upregulating the transcriptional activity of nuclear factor erythroid-2-related factor 2 and the expression of antioxidant genes. Furthermore, MSH synergistically attenuated the expression of proinflammatory cytokines in CCl4-injured liver and lipopolysaccharide-stimulated RAW 264.7 cells. Taken together, these findings suggest that MSH has the potential to prevent acute liver damage by effectively suppressing oxidative stress and inflammation.

Activatable Raman Probes Utilizing Enzyme-Induced Aggregate Formation for Selective Ex Vivo Imaging

Detecting multiple enzyme activities simultaneously with high spatial specificity is a promising strategy to investigate complex biological phenomena, and Raman imaging would be an excellent tool for this purpose due to its high multiplexing capabilities. We previously developed activatable Raman probes based on 9CN-pyronins, but specific visualization of cells with target enzyme activities proved difficult due to leakage of the hydrolysis products from the target cells after activation. Here, focusing on rhodol bearing a nitrile group at the position of 9 (9CN-rhodol), we established a novel mechanism for Raman signal activation based on a combination of aggregate formation (to increase local dye concentration) and the resonant Raman effect along with the bathochromic shift of the absorption, and utilized it to develop Raman probes. We selected the 9CN-rhodol derivative 9CN-JCR as offering a suitable combination of increased stimulated Raman scattering (SRS) signal intensity and high aggregate-forming ability, resulting in good retention in target cells after probe activation. By using isotope-edited 9CN-JCR-based probes, we could simultaneously detect β-galactosidase, γ-glutamyl transpeptidase, and dipeptidyl peptidase-4 activities in live cultured cells and distinguish cell regions expressing target enzyme activity in Drosophila wing disc and fat body ex vivo.

Improvement in salt-tolerance of Aspergillus oryzae γ-glutamyl transpeptidase via protein chimerization with Aspergillus sydowii homolog

γ-Glutamyl transpeptidase is one of the key enzymes involved in glutamate production during high-salt fermentation of soy sauce and miso by koji mold, Aspergillus oryzae. However, the activity of γ-glutamyl transpeptidase from A. oryzae (AOggtA) is markedly reduced in the presence of NaCl, thus classifying it as a non-salt-tolerant enzyme. In contrast, the homologous protein from the xerophilic mold, A. sydowii (ASggtA) maintains its activity under high-salt conditions. Therefore, in this study, a chimeric enzyme, ASAOggtA, was designed and engineered to improve salt-tolerance in AOggtA by swapping the N-terminal region, based on sequence and structure comparisons between salt-tolerant ASggtA and non-salt-tolerant AOggtA. The parental AOggtA and ASggtA and their chimera, ASAOggtA, were heterologously expressed in A. oryzae and purified. The chimeric enzyme inherited the superior activity and stability from each of the two parent enzymes. ASAOggtA showed > 2-fold greater tolerance than AOggtA in the presence of 18% NaCl. In addition, the chimera showed a broader range of pH stability and greater thermostability than ASggtA. AOggtA and ASAOggtA were sy over the range pH 3.0 to pH 10.5. Thermal stability was found to be in the order AOggtA (57.5 °C, t1/2 = 32.5 min) > ASAOggtA (55 °C, t1/2 = 20.5 min) > ASggtA (50 °C, t1/2 = 12.5 min). The catalytic and structural characteristics indicated that non-salt-tolerant AOggtA would not undergo irreversible structural changes in the presence of NaCl, but rather a temporary conformational change, which might result in reducing the substrate binding and catalytic activity, on the basis of kinetic properties. In addition, the chimeric enzyme showed hydrolytic activity toward L-glutamine that was as high as that of AOggtA. The newly-designed chimeric ASAOggtA might have potential applications in high-salt fermentation, such as miso and shoyu, to increase the content of the umami-flavor amino acid, L-glutamate.

Immobilization of E. coli expressing γ-glutamyltranspeptidase on its surface for γ-glutamyl compound production

An Escherichia coli strain expressing γ-glutamyltranspeptidase on its extracellular surface using the Met1 to Arg232 fragment of YiaT of E. coli as an anchor protein was immobilized with alginate for repeated use. Measurement of γ-glutamyltranspeptidase activity of the immobilized cells was performed repeatedly at pH 8.73 and 37 °C for 10 days using γ-glutamyl-p-nitroanilide in the presence of 100 mM CaCl2 and 3% NaCl with and without glycylglycine. Even after the 10th day, the enzyme activity did not decrease from the initial levels. The production of γ-glutamylglutamine from glutamine using the immobilized cells was performed repeatedly at pH 10.5 and 37 °C for 10 days in the presence of 250 mM glutamine, 100 mM CaCl2, and 3% NaCl. Sixty-four % of glutamine was converted to γ-glutamylglutamine in the first cycle. While repeating the production 10 times, the surface of the beads gradually became covered with white precipitate, and the conversion efficiency gradually decreased, but 72% of the initial value still remained even at the 10th measurement.

Alteration of Glutathione and Antioxidant Status with Exercise in Unfed and Refed Rats

The influences of food deprivation and refeeding on glutathione (GSH) status, antioxidant enzyme activity and lipid peroxidation in response to an acute bout of exercise were investigated in the liver and skeletal muscles of male Sprague-Dawley rats randomly divided into three groups: starved for 48 h without refeeding; starved for 48 h and refed for 24 or 48 h. Half of each group of rats was exercised on a treadmill until exhaustion and killed immediately, whereas the other half group was killed at rest. Food-deprived rats had significantly lower liver GSH concentration and GSH:glutathione disulfide (GSSG) ratio. Malondialdehyde concentrations in the liver and skeletal muscle were both higher in the starved than in the refed rats (P < 0.05). Refed rats had significantly greater liver GSH level, γ-glutamylcysteine synthetase and glucose 6-phosphate dehydrogenase activities and plasma insulin concentration than unfed rats. Exercised 24- and 48-h refed rats had 27% and 31% lower liver GSH (P < 0.05), respectively, and a 21% lower GSH:GSSG ratio (P < 0.05) than their rested counterparts. Plasma insulin concentrations were significantly lower, whereas glucagon concentrations were greater in the exercised than in the rested rats. Muscle GSH concentration was significantly lower in the food-deprived than in the refed rats (P < 0.05) but was unaffected by exercise. Exercised 24-h refed rats had significantly elevated muscle GSSG concentration compared with rested rats, along with a higher GSH peroxidase and a lower γ-glutamyltranspeptidase activity (P < 0.05). These data indicate that both food deprivation-refeeding and exhaustive exercise influence liver and skeletal muscle glutathione status and that these changes may be controlled by hepatic glutathione synthesis and release due to hormonal stimulation.

Evaluation of Cottonseed Meal as an Alternative to Fish Meal in Diet for Juvenile Asian Red-Tailed Catfish Hemibagrus wyckioides.

A 10-week trial was performed to investigate the effects of replacing fishmeal with cottonseed meal (CSM) on the growth rate, protein metabolism, and antioxidant response of Asian red-tailed catfish Hemibagrus wyckioides. Five isonitrogenous and isocaloric diets (C0, C8.5, C17.2, C25.7, and C34.4) were prepared to contain 0%, 8.5%, 17.2%, 25.7%, and 34.4% CSM replacing fishmeal, respectively. The weight gain, daily growth coefficient, pepsin, and intestinal amylase activities initially increased and then decreased with the raising dietary CSM levels; the highest values were observed in the C17.2 group (P < 0.05). However, feed cost exhibited the opposite trend. With the increasing dietary CSM levels, the protein efficiency ratio and intestinal trypsin activity decreased but feed conversion rate increased gradually; while no differences were observed among the C0, C8.5, and C17.2 groups (P > 0.05). Dietary CSM inclusion regardless of levels increased the plasma growth hormone level as well as hepatic aspartate aminotransferase (AST) and γ-glutamyl transpeptidase activities but decreased the plasma glutamate dehydrogenase and AST activities (P < 0.05). With the increasing dietary CSM levels, the plasma alkaline phosphatase (AKP) and hepatic superoxide dismutase activities decreased but malondialdehyde content increased gradually, while no differences were observed among the C0, C8.5, and C17.2 groups (P > 0.05). The plasma immunoglobulin M content and hepatic glutathione reductase activity initially increased but then decreased with the raising dietary CSM levels; the highest values were found in the C17.2 group. These results indicated that dietary CSM inclusion level up to 17.2% improved the growth rate, feed cost, digestive enzyme activity, and protein metabolism without compromising antioxidant capacity of H. wyckioide, whereas these parameters were depressed by further inclusion of CSM. CSM is a potentially cost-effective alternative plant protein source in diet of H. wyckioide.

First Example of the Extracellular Surface Expression of Intrinsically Periplasmic Escherichia coli γ-Glutamyltranspeptidase, a Member of the N-Terminal Nucleophile Hydrolase Superfamily, and the Use of Cells as a Catalyst for γ-Glutamylvalylglycine Production.

Although the purified Escherichia coli γ-glutamyltranspeptidase has much higher transpeptidation activity than hydrolysis activity, almost all γ-glutamyltranspeptidase activity is hydrolysis activity in vivo, that is when measured using the whole cells. By using the Met1 to Arg232 fragment of E. coli YiaT or the CapA of Bacillus subtilis subsp. Natto as an anchor protein, we succeeded in expressing E. coli γ-glutamyltranspeptidase on the extracellular surface of the cells, and these cells showed higher transpeptidation activity than hydrolysis activity in the presence of NaCl. Furthermore, E. coli cells overexpressing γ-glutamyltranspeptidase without an anchor from the T5 promoter maintained γ-glutamyltranspeptidase on the extracellular surface of the cells immediately after being harvested from the culture medium, but the enzyme was released from the extracellular surface of the cells subsequently in the absence of NaCl. Using these cells expressing γ-glutamyltranspeptidase on the extracellular surface, γ-Glu-Val-Gly, a kokumi compound, was successfully produced.

Ameliorative Effects of Vanadyl Sulfate on Some Biochemical Parameters of Experimental Diabetic Rat Kidneys

Diabetes mellitus (DM), closely related to diabetic nephropathy, is one of the major public health problems worldwide. Today, with the increasing understanding of the underlying pathophysiology of DM, new oral anti-diabetic treatment strategies are being developed. Vanadium is a transition element that is widely distributed in nature, and its oral administration has been reported to improve DM in humans and a variety of diabetic animal models. The purpose of the research is to explore the effect of vanadyl sulfate (VS) administration on the different enzyme activities associated with kidney injury in streptozotocin- (STZ) induced diabetic rats. Male rats were assigned into groups as follows: untreated control, control animals given VS (100 mg/kg), diabetic (a single dose of intraperitoneal STZ, 65 mg/kg), and diabetic + VS (same dose) group. VS was administered orally for 60 days after the induction of diabetes. On the 60th day of experiment, kidney samples were taken for analysis. According to the data obtained from the biochemical analysis, the activities of transaminases, alkaline phosphatase, carbonic anhydrase, and γ-glutamyl transpeptidase decreased, whereas superoxide dismutase activity elevated in the kidney tissue of VS treated hyperglycemic animals. The results suggested that VS improved the diabetic renal injury, probably by VS insulin-mimic and antioxidant behavior through decreased oxidative stress and increased antioxidant capacity. Therefore, vanadyl sulfate might be used as a potential oral anti-diabetic compound in the treatment of the diabetic nephropathy, and as an important control for elevated blood glucose levels in the diabetic state.

Association of serum brain-derived neurotrophic factor with hepatic enzymes, AST/ALT ratio, and FIB-4 index in middle-aged and older women.

Substantial evidence suggests an important role of liver function in brain health. Liver function is clinically assessed by measuring the activity of hepatic enzymes in the peripheral blood. Brain-derived neurotrophic factor (BDNF) is an important regulator of brain function. Therefore, we hypothesized that blood BDNF levels are associated with liver function and fibrosis. To test this hypothesis, in this cross-sectional study, we investigated whether serum BDNF concentration is associated with liver enzyme activity, aspartate aminotransferase (AST)/ alanine aminotransferase (ALT) ratio, and fibrosis-4 (FIB-4) index in middle-aged and older women. We found that serum BDNF level showed a significant positive association with ALT and γ-glutamyltranspeptidase (GGT) activity and negative association with FIB-4 index, and a trend of negative association with the AST/ALT ratio after adjustment for age. Additionally, these associations remained statistically significant even after adjustment for body mass index (BMI) and fasting blood glucose level. These results demonstrate associations of serum BDNF levels with liver enzymes and hepatic fibrosis-related indices, which may underlie liver-brain interactions.

A water-soluble fluorescent probe for real-time visualization of γ-glutamyl transpeptidase activity in living cells

γ-glutamyl transpeptidase (GGT) is a kind of cell-surface enzyme that is overexpressed in many cancer cells. It is of great significance to develop an ideal tool for the diagnosis of GGT-rich cancer cells. Here, we reported a simple-structured but effective imaging probe for the detection of GGT activity. In the presence of GGT, the γ-glutamyl linkage could be cleaved specifically to produce amino-substituted product, resulting in significant fluorescence enhancement at 578 nm. Moreover, we successfully employed the probe to monitor GGT activity in HepG2 cells. We envisaged that such a simple but effective imaging tool could improve the practical applications for bioimaging.

Sensitive sensing of alkaline phosphatase and γ-glutamyltranspeptidase activity for tumor imaging.

Alkaline phosphatase (ALP) and γ-glutamyltranspeptidase (GGT) are regarded as two important biomarkers in several human cancers. There are many probes for ALP or GGT detection but no probe was reported to sense the activity of both ALP and GGT in vitro or in vivo. Herein, a bioluminescent probe P-Bz-Luc was designed for realizing sensitive and specific tumor imaging via the co-cleavage of ALP and GGT. In solution experiments showed that an excellent linear relationship was found between the bioluminescence signal of the P-Bz-Luc solution and the enzyme concentration at limits of detection of 0.172 for ALP and 0.634 U L-1 for GGT. Eventually, the BL probe P-Bz-Luc was successfully applied for sensitive imaging of the ALP and GGT co-overexpressed fLuc-231 breast cancer cells and tumors, with the help of the coordination cleavage of P-Bz-Luc by ALP and GGT.

Changes in the physicochemical composition of Auricularia auricula during growth stages and control of endogenous formaldehyde

To maintain quality and reduce endogenous formaldehyde content in the edible fungus, Auricularia auricula, the critical period of formaldehyde generation was identified and controlled. In this study, different growth stages of A. auricula were used to assess the nutritional composition (polyphenol monomers, 5′-nucleotides, organic acids and water-soluble vitamins) and related enzyme activities; the correlation between quality and endogenous formaldehyde content was also analysed. The results showed that formaldehyde was positively correlated with soluble protein and melanin levels; it was negatively correlated with the total sugar and flavonoid contents. Among the four growth stages, ripe A. auricula (RAA) had the highest endogenous formaldehyde content. The activities of γ-glutamyl transpeptidase (GGT) and l-cysteine sulphoxide lyase (C-S lyase) were highest in RAA; they promoted endogenous formaldehyde production. RAA is a critical period for interventions to reduce endogenous formaldehyde contents. Experiments to distinguish the effects of 10 exogenous substances on GGT and C-S lyase indicated that cysteine (Cys) and glutamic acid (Glu) inhibited the production of endogenous formaldehyde by 17.13% and 14.89%, respectively. This study provides a new perspective for controlling safety during the growth of edible fungi and maintaining the quality of processed products.

Prospective evaluation of 5 urinary biomarkers as predictors of acute kidney injury in nonazotemic, hospitalized dogs.

BackgroundEarly recognition of acute kidney injury (AKI) is hindered by current definitions and use of traditional, insensitive markers.Hypothesis/ObjectivesUrinary (u) activity of γ‐glutamyl transpeptidase (GGT) and alkaline phosphatase (ALP), and concentrations of heat‐shock protein 70 (HSP70) and interleukins (ILs) ‐6 and ‐18, are predictive biomarkers for AKI and survival.AnimalsNonazotemic, hospitalized dogs (n = 118) and healthy controls (n = 20).MethodsA prospective observational study. Nonazotemic dogs at risk of AKI were recruited and their urinary biomarker concentrations were measured at presentation. Serum creatinine (sCr) and symmetric dimethylarginine (sSDMA) were measured daily until discharge/death.ResultsThe overall case fatality rate was 18.6%. Fifteen dogs (12.7%) developed AKI, which was associated with death (relative risk, 3.2; 95% confidence interval [CI], 1.57‐6.55). All 5 urinary biomarkers were significantly higher in hospitalized dogs compared to controls, with minimal overlap. uHSP70/uCr, uGGT/uCr, and uIL‐6/uCr at presentation were higher in dogs which later developed AKI. Areas under the receiver operator characteristic curve (AUROC) (95% CI) for the 3 biomarkers as predictors of AKI were 0.67 (0.51‐0.83), 0.68 (0.55‐0.81), and 0.78 (0.65‐0.91), respectively. When they were categorically classified as elevated/normal, each additional elevated biomarker increased the odds for AKI (OR, 2.83; 95% CI, 1.23‐6.52, P = .01). Agreement between sCr and sSDMA was poor (Cohen's kappa = .071). The AUROC of SDMA at presentation for AKI prediction was 0.73 (0.51‐0.95).Conclusions and Clinical ImportanceKidney injury was common, irrespective of subsequent worsening of azotemia or death. The predictive value of individual urinary biomarkers was reduced by moderate sensitivities and specificities. SDMA showed moderate discriminatory utility for AKI prediction, and often displayed discordant results with sCr.

Enzymatic and fecundity evaluation of Fasciola hepatica exposed to different doses of γ- irradiation in Ethiopian sheep

The upshot of γ-irradiated Fasciola hepatica infection on the activity of plasma glutamate dehydrogenase (GLDH) and γ-glutamyl transpeptidase (GGT) was evaluated in 36 sheep infected with a single dose of 30, 60, 120, and 240 grays and those kept as negative and positive control over 17 weeks. During this period, serum and faecal samples, as well as body weight gains, were taken at weekly intervals. Furthermore, the effects of the irradiation dose (500) for oral vaccination and on the recovery of adult flukes were assessed following primary infection. Eggs were first detected in the faeces of infected sheep on week 8 post-infection. The parasite viability was severely affected by doses of γ-irradiation of 120 Gy or 240 Gy. In the aforementioned doses, relatively low numbers of mature flukes of about 60 (17.1%) and 38 (10.8%) were recovered than the control group, respectively. The sensitized lambs also showed less hepatic damage compared with the controls as indicated by lower levels of the serum enzyme glutamate dehydrogenase and γ- glutamyl transferase significant body weight loss was observed between weeks 6 and 8 post-infection followed by a steady increase of the mean weight of infected animals across time. In conclusion, vaccination of sheep with γ irradiated metacercariae of F. hepatica appeared to affect the number and development of the fluke population resulting in reduced hepatic damage during migration, reduced fecundity after patency, as measured by worm and egg counts, levels of serum glutamate dehydrogenase and γ- glutamyl transferase.

Estrogen Deficiency Aggravates Fluoride-Induced Liver Damage and Lipid Metabolism Disorder in Rats.

Estrogen exerts essential role in liver metabolism, and its deficiency is frequently accompanied by a series of metabolic disorder diseases. To investigate the role of estrogen deficiency in fluorine ions (F-) induced liver injury, the ovariectomy (OVX) rat models were performed by surgically removing the ovaries, and the rats from OVX and non-OVX models were exposed to differential dose of F- (0, 25, 50 and 100mg/L) in drinking water for 90days. The liver morphological structure was evaluated by hematoxylin-eosin staining. Proliferation ability of hepatocytes was evaluated by 5-bromo-2-deoxyuridine (BrdU) assay. And distribution of lipid droplets in liver tissue was observed via oil red O staining. In addition, the liver function and lipid metabolism parameters in serum were detected by commercial kits. Results showed that F- induced hepatocytes morphological damage and inhibited the proliferation ability of hepatocytes; estrogen deficiency exacerbated these changes. The deposition of lipid droplets in the liver tissue was multiplicative with increased F- dose, especially after estrogen deficiency. In addition, F- exposure increased (P < 0.05 or P < 0.01) serum aminotransferase (ALT), aminotransferase (AST), alkaline phosphatase (ALP), and γ-glutamyl transpeptidase (γ-GT) activities and total bilirubin (T-bil) level; meanwhile, serum triglyceride (TG) and cholesterol (TC) levels were also elevated (P < 0.05 or P < 0.01). F--induced liver function and lipid metabolism indexes were further increased (P < 0.05 or P < 0.01) in the state of estrogen deficiency. In conclusion, estrogen deficiency aggravated F--induced liver damage and lipid metabolism disorder.

Simultaneous sensing γ-glutamyl transpeptidase and alkaline phosphatase by robust dual-emission carbon dots

Rapid, convenient, sensitive and simultaneous detection of distinct enzymes is urgently needed for diagnosis, therapeutics and prognostic of related diseases. Here, a new strategy for simultaneous monitoring γ-glutamyl transpeptidase (GGT) and alkaline phosphatase (ALP) activity has been fabricated based on dual-emission carbon dots (CDs). CDs were prepared by solvothermal treatment of Actinidia chinensis, which presents two fluorescent emissions at 471 nm (blue channel) and 671 nm (red channel). GGT and ALP activity can be detected based on inner filter effect (IFE) and static quenching effect (SQE) of blue and red channels of CDs, respectively. Linear ranges were 2.5–90 U L−1 and 5–200 U L−1, and limit of detection (LOD) were 0.71 U L−1 and 1.2 U L−1 for GGT and ALP, respectively. Developed CDs can monitor GGT and ALP activity in human serum samples with satisfied recoveries (99.3%–108.6% for GGT, 98.4%–105.4% for ALP). Furthermore, the combination of CDs to sense GGT and ALP activity with OR logic gate can predict human health status. The design and application of dual-emission CDs can also be extended as promising tools to detect multianalytes using different channel signals.

Alterations in acid-base balance, blood gases, and hematobiochemical profiles of whole-blood and thoracic fluid in goats with contagious caprine pleuropneumonia.

Background and Aim:Contagious caprine pleuropneumonia (CCPP) is a highly contagious and fatal disease affecting goats and some wild ruminants. It is a cause of major economic losses in the goat industry in Africa, Asia, and the Middle East. This study aimed to investigate the acid-base balance, blood gases, and hematobiochemical profiles of whole-blood and fluid collected from the thoracic cavity in goats with CCPP.Materials and Methods:Fifty-five goats suffering from weight loss, anorexia, dyspnea, polypnea, cough, and nasal discharges due to CCPP were studied. Twenty-five healthy goats were used as controls. Diseased animals were enrolled in this study based on a positive serological latex agglutination test (LAT) that confirmed the detection of Mycoplasma capricolum subsp. capripneumoniae. The control goats were enrolled based on a negative result of the LAT.Results:Compared with a mean value of 7.38±0.04 in controls, the pH in the diseased group was 7.41±0.05. The blood pressure of carbon dioxide (PCO2), pressure of oxygen (PO2), base excess (BE), bicarbonate (HCO3), total carbon dioxide (TCO2), and saturation of oxygen (SO2) were lower in goats with CCPP than in controls. However, the anion gap (AnGap) was higher in the diseased goats than in the healthy ones. Compared with the levels in blood samples, the thoracic fluid PCO2, PO2, BE, and SO2 were higher while pH, HCO3, TCO2, and AnGap were lower. Compared with the findings in healthy goats, hematological alterations included significant increases in white blood cells and neutrophils, and a significant decrease in the red blood cell count. In the thoracic fluid, neutrophilic leukocytosis was a remarkable finding. The serum concentrations of globulin, blood urea nitrogen, and glucose, and the activities of aspartate aminotransferase (AST) and g-glutamyl transpeptidase (GGT) increased significantly compared with those in controls. In contrast, serum concentrations of albumin, calcium, and magnesium, and the activity of alkaline phosphatase (ALP) decreased significantly compared with those of healthy animals. The activities of ALP, AST, GGT, and creatine kinase and the concentration of phosphorus were higher in thoracic fluid than the serum values in the diseased group.Conclusion:When compared with the healthy controls, goats with CCPP have metabolic acidosis. Compared with the levels in healthy goats, the blood PCO2, PO2, BE, HCO3, TCO2, and SO2 are low in goats with CCPP; however, the AnGap is higher in diseased goats.

Megalin-mediated albumin endocytosis in renal proximal tubules is involved in the antiproteinuric effect of angiotensin II type 1 receptor blocker in a subclinical acute kidney injury animal model

BackgroundTubule-interstitial injury (TII) is one of the mechanisms involved in the progression of renal diseases with progressive proteinuria. Angiotensin II (Ang II) type 1 receptor blockers (ARBs) have been successfully used to treat renal diseases. However, the mechanism correlating treatment with ARBs and proteinuria is not completely understood. The hypothesis that the anti-proteinuric effect of losartan is associated with the modulation of albumin endocytosis in PT epithelial cells (PTECs) was assessed. MethodsWe used a subclinical acute kidney injury animal model (subAKI) and LLC-PK1 cells, a model of PTECs. ResultsIn subAKI, PT albumin overload induced TII development, measured by: (1) increase in urinary lactate dehydrogenase and γ-glutamyltranspeptidase activity; (2) proteinuria associated with impairment in megalin-mediated albumin reabsorption; (3) increase in luminal and interstitial space in tubular cortical segments. These effects were avoided by treating the animals with losartan, an ARB. Using LLC-PK1 cells, we observed that: (1) 20 mg/mL albumin increased the secretion of Ang II and decreased megalin-mediated albumin endocytosis; (2) the effects of Ang II and albumin were abolished by 10−8 M losartan; (3) MEK/ERK pathway is the molecular mechanism underlying the Ang II-mediated inhibitory effect of albumin on PT albumin endocytosis. ConclusionOur results show that PT megalin-mediated albumin endocytosis is a possible target during the treatment of renal diseases patients with ARB. General significanceThe findings obtained in the present work represents a step forward to the current knowledge on about the role of ARBs in the treatment of renal disease.