Γ-glutamyl Transpeptidase Activity Research Articles

Dual-channel near-infrared fluorescent probe for real-time tracking of endogenous γ-glutamyl transpeptidase activity.

A curcuminoid difluoroboron-based tumor-targeting GGT-activatable fluorescent probe (Glu-DFB-biotin) is designed, which has remarkable characteristics of dual-channel and light-up near-infrared (NIR) fluorescence. Furthermore, Glu-DFB-biotin can differentiate hepatic cancer cells from normal cells and is successfully employed for real-time tracking of endogenous GGT activity in living cells.

Bioluminescence probe for γ-glutamyl transpeptidase detection in vivo

To detect γ-Glutamyl Transpeptidase (GGT) activity in vitro and in vivo, a bioluminescence probe with high sensitivity and specificity was well designed and synthesized. This probe can be recognized by GGT and release strong bioluminescence with its further reaction with luciferase. The performance of this probe was demonstrated in vitro and in cells. Finally, we applied the probe for detection of GGT activity in xenograft model.

Anti-tumor activities of macromolecular fractions of fresh gecko in vivo and their induction of Bel-7402 cell differentiation

ObjectiveTo investigate the anti-tumor effect of macromolecular fractions of fresh gecko (M-AG) in vivo and their differentiation-inducing activity in Bel-7402 cells in vitro. MethodsAn H22 hepatocarcinoma-bearing mouse model was used to evaluate the anti-tumor activity of M-AG samples. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was applied to analyze cell viability. Cell morphology was observed by phase contrast microscopy. The quantity of the alpha-fetoprotein was detected by a radioimmunoassay. Chromatometry was used to assay the albumin quantity. Activities of alkaline phosphatase and γ-glutamyl trans-peptidase were measured by biochemical methods. Finally, western blotting was applied to assess proteins in the mitogen-activated protein kinase (MAPK) signaling pathway. ResultsMacromolecular fractions of fresh gecko exerted a significant anti-tumor effect in mice. The inhibition rate of tumor growth was 63% in the moderate M-AG dose group. Cells treated with M-AG displayed a differentiated state. The treatment lowered alpha-fetoprotein secretion and significantly decreased the activities of γ-glutamyl trans-peptidase and alkaline phosphatase in Bel-7402 cells. In contrast, M-AG increased the amount of albumin in the cell culture medium. All biochemical indices demonstrated that M-AG induced Bel-7402 cell differentiation. Western blotting showed no changes in the quantities of extracellular signal-regulated kinase (ERK) 1/2, p38 MAPK, or c-Jun N-terminal protein kinase 1/2. However, M-AG significantly activated the phosphorylation of ERK1/2 in a dose-dependent manner. In addition, M-AG had no significant influence on the expression of nuclear factor-kappa B. ConclusionMacromolecular fractions of fresh gecko has an anti-tumor activity in H22 hepatocarcinoma-bearing mice in vivo and inhibits Bel-7402 cell proliferation in vitro by inducing cell differentiation related to activation of ERK1/2.

Functional validation of novel MKS3/TMEM67 mutations in COACH syndrome

COACH syndrome is an autosomal recessive developmental disorder, a subtype of Joubert syndrome and related disorders, characterized by cerebellar vermis hypoplasia, oligophrenia, ataxia, coloboma, and hepatic fibrosis. Although mutations in TMEM67 (transmembrane protein 67)/MKS3 (Meckel-Gruber syndrome, type 3) were reported to cause COACH syndrome, this causality has not verified by functional studies. In a 20-year-old Korean man, we found cerebellar ataxia, isolated elevation in serum γ-glutamyl transpeptidase (γ-GTP) activity, oligophrenia, the molar tooth sign (MTS) in the brain MR images and congenital hepatic fibrosis (CHF). Two novel compound heterozygous mutations were found in TMEM67 in the patient: i) missense mutation (c.395 G > C and p.Gly132Ala) in exon 3, and ii) deletion in exon 26 (c.2758delT and p.Tyr920ThrfsX40). Western blotting showed that the p.Tyr920ThrfsX40 mutation accelerates turnover of the TMEM67 protein. Although wild-type human TMEM67 RNA rescued phenotypes of zebrafish embryos injected with anti-sense oligonucleotide morpholinos against tmem67, the two human TMEM67 RNAs individually harboring the two mutations did not. Finally, Wnt signaling, but not Hedgehog signaling, was suppressed in tmem67 morphants. To the best of our knowledge, this is the first report verifying the causality between COACH syndrome and TMEM67, which will further our understanding of molecular pathogenesis of the syndrome.

Postconditioning attenuates bile duct cell damage by apoptosis from liver ischemia-reperfusion injury in rats

Objective To study the protective effect of postconditioning on bile duct cell damage in the form of apoptosis during liver ischemia-reperfusion (IR) injury by observing the expression of B cell lymphoma/leukemia-2 (bcl-2) and B cell lymphoma/leukemia-2 associated X protein (bax) on bile ducts. Methods Thirty healthy male SD rats were randomly divided into three groups: sham-operation (SO) group, IR group and ischemic postconditioning (IPO) group. IPO was achieved by 6 brief reperfused/ischemia cycles before the persistent reperfusion procedure. The samples of blood and hepatic tissue of all groups were taken after experiment. The activity of γ-glutamyl transpeptidase (GT) in serum was determined. The expression of bcl-2 and bax on bile ducts was detected by immunohistochemistry. The apoptosis index on bile ducts was observed by TdT-mediated dUTP nick end labeling (TUNEL) methods. Results The activity of γ-GT was lower in IPO group [(26.30±3.32) U/L] than that in IR group [(34.64±3.28) U/L] with the difference being statistically significant (P=0.001). As compared with the IR group, the expression of bcl-2 on bile ducts was up-regulated [(77.94±5.25)% vs. (42.67±3.73)%], and that of bax down-regulated [(70.10±4.27)% vs. (90.17±2.12)%] in IPO group, with the difference being statistically significant (P=0.000). As compared with IR group, the apoptosis index in IPO group was decreased [(58.90±2.75)% vs. (79.31±1.12)%, P=0.000], and the pathologically morphologic damages of the bile duct cells were attenuated. Conclusion In the process of liver IR, the postconditioning can promote the expression of bcl-2 and inhibit the expression of bax on bile duct, which may withstand the happening of the bile duct cell apoptosis, thus attenuating the bile duct damage caused by liver IR injury. Key words: Reperfusion injury; Postconditioning; Bile duct; Apoptosis; B cell lymphoma/leukemia-2; B cell lymphoma/leukemia-2 associated X protein

Rice bran phytic acid protects against methotrexate -induced oxidative stress and acute liver injury in rats

Methotrexate is an effective and extensively used chemotherapeutic agent to treat range of malignancies, but its therapeutic use is limited because of dose dependent hepatotoxicity. Several published reports advocate that supplementation with antioxidant can influence methotrexate induced acute liver injury. Twenty four adult male rats (aged 56 days and weighted 138±8.8g) were randomly assigned into four equal groups (control and three treated groups), first group as control group given normal saline, a second single injection 20mg /kg B.W. i.p. of methotrexate, third group given15mg/kg B.W. of phytic acid, then given single injection 20mg /kg B.W. i.p. of methotrexate and fourth group given 15mg/kg B.W. of phytic acid for 10 days, and male rats were sacrificed. Blood and liver, subcellular fluid was obtained to assess subcellular activity of Alanine aminotranferease (ALT), Aspartate aminotransferase (AST), reduced glutathione (GSH), glutathione reductase, glutathione-S-transferase (GST), glutathione peroxidase (GPx), catalase, quinone reductase (QR), superoxide dismutase (SOD), γ-glutamyl transpeptidase (GGT), lactate dehydrogenase (LDH), lipid peroxidation, glucose-6-phosphate dehydrogenase (G6PD) and total protein. The results of liver injury rats (T) showed significant increased activity of ALT, AST, GGT, LDH, decreased GSH, GR, GST, GPx, CAT, QR, SOD, (p<0.05) when compared with the control, and histological findings further supported the protective effects of phytic acid against methotrexate induced acute liver injury. phytic acid (D) therapy moderated liver damage and normalized the activities of all antioxidant enzymes. In conclusion, present study demonstrate that oxidative stress and liver injury are closely associated with methotrexate induced toxicity and phytic acid shows the protective efficacy against methotrexate induced acute liver injury possibly via attenuating the oxidative stress and inflammatory response.

Adrenergic regulation during acute hepatic infection with Entamoeba histolytica in the hamster: involvement of oxidative stress, Nrf2 and NF-KappaB.

Oxidative stress and transcriptional pathways of nuclear factor erythroid 2-related factor 2 (Nrf2) and nuclear factor kappa-B (NF-κB) are critically involved in the etiopathology of amebic liver abscess (ALA). In this work, we studied the relationship between the adrenergic nervous system and ALA in the hamster. ALA was visible at 12 h of infection. While 6-hydroxidopamine (6-OHDA) decreased infection, propranolol (β-adrenergic blocker) treatment was associated with less extensive liver damage, and phentolamine treatment (α-adrenergic blocker) significantly reduced ALA compared to 6-OHDA and propranolol. Serum enzymatic activities of alanine aminotransferase (ALT) and γ-glutamyl transpeptidase (γ-GTP) were increased at 12 h post-infection. Chemical denervation and α and β-adrenergic blockers decreased ALT to normal levels, while 6-OHDA and propranolol showed a trend to decrease γ-GTP but phentolamine significantly reduced γ-GTP. Amebic infection increased oxidized glutathione (GSSG) and decreased both reduced glutathione (GSH) and the GSH/GSSG ratio. Propranolol and 6-OHDA showed a tendency to decrease GSSG. However, GSH, GSSG and GSH/GSSG returned to normal levels with phentolamine. Furthermore, amebic infection increased pNF-κB and interleukin-1β (IL-1β), and showed a tendency to decrease hemoxigenase-1 (HO-1), but not Nrf2. Chemical denervation showed a trend to decrease pNF-κB and IL-1β, and neither Nrf2 nor HO-1 increased significantly. In addition, NF-κB and IL-1β were attenuated by propranolol and phentolamine treatments, although phentolamine showed significant overexpression of Nrf2 and HO-1. This suggests that the adrenergic system may be involved in oxidative stress and in modulation of the Nrf2 and NF-κB pathways during ALA development.

An efficient two-photon fluorescent probe for measuring γ-glutamyltranspeptidase activity during the oxidative stress process in tumor cells and tissues.

Oxidative stress, a disturbance in the balance between oxidant/antioxidant ratios, is associated with cancer, aging, inflammation, neurodegenerative diseases and other conditions. γ-Glutamyltranspeptidase (GGT) is a redox-related enzyme that plays a key role in mitigating the effects of oxidative stress by maintaining cellular glutathione (GSH) metabolism and homeostasis. Therefore, oxidative stress will upregulate the intracellular GGT level. To better understand the major pathophysiological resist mechanism to oxidative injury in mediating many disease states, we designed and synthesized a novel two-photon (TP) fluorescent turn-on probe, Np-Glu, for GGT detection and bioimaging. Under the optimized conditions, Np-Glu exhibited remarkable fluorescence enhancement (150-fold), good selectivity and high sensitivity (LOD is 0.033 U L-1), with a wide linear concentration range (0-50 U L-1). More importantly, the probe Np-Glu was successfully applied in one-photon and TP fluorescence imaging of GGT activity in an oxidative stress model in living cells and tissues, suggesting Np-Glu as an ideal indicator for clinical and biological samples.

Utility of urinary alkaline phosphatase and γ-glutamyl transpeptidase in diagnosing acute kidney injury in dogs

The diagnostic utility of urinary alkaline phosphatase (uALP) and γ-glutamyl transpeptidase (uGGT) activities in naturally occurring acute kidney injury (AKI) was investigated in a heterogeneous group of dogs. The study included client-owned dogs with AKI (n = 32), chronic kidney disease (CKD, n = 13), lower urinary tract infection (LUTI, n = 15) and healthy controls (n = 24). uGGT and uALP activities were normalised to urinary creatinine (uCr) concentrations (uGGT/uCr and uALP/uCr, respectively). uALP/uCr and uGGT/uCr were positively and significantly correlated (r = 0.619, P < 0.001), and differed significantly (P ≤ 0.001) among groups, as well as between AKI and LUTI or CKD groups (P < 0.05), but not between the AKI and control groups. Areas under the receiver operator characteristics (ROC) curve for uALP/uCr and uGGT/uCr as predictors of AKI were 0.75 and 0.65, respectively, with optimal cut-off points showing poor to moderate sensitivity (59% for uALP/uCr and 79% for uGGT/uCr) and specificity (59% for uALP/uCr and 75% for uGGT/uCr). Higher cut-off points, with 90% specificity, showed low sensitivity (41% for both uALP/uCr and uGGT/uCr). In conclusion, uALP/uCr is superior to uGGT/uCr as a marker of AKI, but both uGGT/uCr and uALP/uCr have unsatisfactory discriminatory power for diagnosing naturally occurring AKI in dogs and therefore cannot be recommended as sole screening tests for canine AKI. However, both may serve as ancillary, confirmatory, biomarkers for detecting AKI in dogs if appropriate cut-off points with high specificities are used.

Elevated liver enzymes

Elevated liver enzymes are a frequent finding in both symptomatic and asymptomatic patients necessitating further evaluation to clarify the underlying disease. Three different patterns of increased liver enzymes can be defined to allow for a more precise and rational further diagnostic approach. A predominant increase in transaminase activities reflects a disturbance of hepatocellular integrity which can be found in patients with viral hepatitis, genetic liver diseases like Wilson`s disease or hemochromatosis, and drug-induced liver diseases. A second pattern is characterized by high serum alkaline phosphatase and γ-glutamyltranspeptidase activities indicating cholestatic liver diseases. The next important diagnostic measure in this group is an ultrasound study discerning intra- from extrahepatic cholestasis. Intrahepatic cholestatic diseases include primary and secondary sclerosing cholangitis, genetic disturbances of canalicular membrane transporters or drug-induced liver dieseases. Extrahepatic cholestasis involves obstruction of the large bile ducts by gall stones or tumors. The third enzym pattern is defined by a predominant rise in γ-glutamyl transpeptidase which is observed in alcoholic or non-alcoholic fatty liver disease and infiltrating liver diseases. A rise in liver enzymes is not necessarily indicative of a primary hepatic origin. Extrahepatic diseases often cause similarly increased serum activities. In addition even higher values can be observed under normal conditions during pregnancy or in adolescens. Lower values in asymptomatic patients should only be controlled since more than 30% of elevated transaminases spontaneously normalize during follow-up.

Hepatoprotective potential of glyceryl trinitrate against chemically induced oxidative stress and hepatic injury in rats.

Glyceryl trinitrate (GTN) has been used widely as a potent vasodilator to treat heart conditions, such as angina pectoris and chronic heart failure. This study aims to elucidate the effect of exogenous nitric oxide (NO) administration, using GTN, on carbon tetrachloride (CCl4)-induced oxidative stress and liver injury in rats. The results obtained demonstrated that NO generated by the administration of GTN affords protection against CCl4-induced oxidative stress and liver injury. Administration of CCl4 resulted in a significant ( p < 0.001) increase in lipid peroxidation and tissue damage markers (aspartate and alanine transaminase and lactate dehydrogenase) release in serum. Parallel to these changes, CCl4 also caused downregulation of antioxidant enzymes including glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione-S-transferase (GST), and several fold induction in γ-glutamyl transpeptidase (GGT) activity. Subsequent administration of GTN resulted in significant ( p < 0.001) recovery of GSH-metabolizing enzymes in a dose-dependent manner. Further, administration of NO inhibitor, NG-nitro-l-arginine methyl ester (l-NAME), exacerbated CCl4-induced oxidative tissue injury. Overall, the study suggests that GTN might suppress oxidant-induced tissue injury and hepatotoxicity in rats.

Steroid Metabolomic Disease Signature of Nonsyndromic Childhood Obesity.

The profile of urinary steroids as measured by gas chromatography-mass spectrometry defines a subject's "steroidal fingerprint." Here, we clustered steroidal fingerprints to characterize patients with nonsyndromic childhood obesity by "steroid metabolomic signatures." Nonsyndromic obesity is a symptom of different diseases and conditions, some of them will have their own signature. A total of 31 steroid metabolites were quantified by gas chromatography-mass spectrometry, and their excretion rates were z-transformed. Using MetaboAnalyst 3.0, we divided the subjects into 5 distinctive groups by k-means clustering. Steroidal fingerprints and clinical/biochemical data of patients in each cluster were analyzed. A total of 87 obese children (44 females), aged 8.5-17.9 years, were clinically characterized, and their 24-hour urine was collected. Cluster 1 (n = 39, 21 females) had normal steroid profile. Cluster 2 (n = 20, 11 females) showed mild, nonspecific elevation of C19 and C21 steroids, females' resistance to polycystic ovary morphology, and hirsutism. Cluster 3 (n = 7 female), with relative 21-hydroxylase insufficiency, was characterized by partial or full polycystic ovary syndrome. Cluster 4 (n = 4 males), showed markedly elevated C21 steroids and imbalance in the 11β-hydroxysteroid dehydrogenase system, higher insulin, increased frequency of glucose/insulin index more than 0.3, γ-glutamyl transpeptidase activity, systolic blood pressure, and tendency to liver steatosis. Cluster 5 (n = 17, 5 females) had elevated dehydroepiandrosterone and 17-OH-pregnenolone metabolites, suggesting 3β-hydroxysteroid dehydrogenase insufficiency but no clinically unique phenotype. Z-score body mass index values were not significantly different between the clusters. We defined a novel concept of disease-specific steroid metabolomic signature based on urinary steroidal gas chromatography-mass spectrometry. Clustering by software designed for metabolic data analysis reclassified childhood obesity into 5 groups with distinctive signatures; groups require further definition and may require cluster-specific therapeutic strategies.

Rapid Cancer Fluorescence Imaging Using A γ-Glutamyltranspeptidase-Specific Probe For Primary Lung Cancer

BACKGROUND: We set out to examine the activity of γ-glutamyltranspeptidase (GGT) in lung cancer and the validity of γ-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG) for intraoperative imaging of primary lung cancer. METHODS: GGT activities and mRNA expression levels of GGT1 (one of the GGT subtypes) in five human lung cancer cell lines were examined by fluorescence imaging and quantitative reverse transcription polymerase chain reaction. In vivo imaging of an orthotopic A549 xenograft model in nude mice was performed to confirm its applicability to intraoperative imaging. Furthermore, ex vivo imaging of 73 specimens from lung cancer patients were performed and analyzed to calculate the sensitivity/specificity of gGlu-HMRG for lung cancer diagnosis. RESULTS: GGT activities and mRNA expression levels of GGT1 are diverse depending on cell type; A549, H441, and H460 showed relatively high GGT activities and expression levels, whereas H82 and H226 showed lower values. In the in vivo mouse model study, tiny pleural dissemination and hilar/mediastinal lymph node metastasis (less than 1 mm in diameter) were clearly detected 15 minutes after topical application of gGlu-HMRG. In the ex vivo study of specimens from patients, the sensitivity and specificity of gGlu-HMRG were calculated to be 43.8% (32/73) and 84.9% (62/73), respectively. When limited to female patients, never smokers, and adenocarcinomas, these values were 78.9% (15/19) and 73.7% (14/19), respectively. CONCLUSIONS: Although GGT activity of lung cancer cells vary, gGlu-HMRG can serve as an intraoperative imaging tool to detect small foci of lung cancer when such cells have sufficient GGT activity.

The prolonged effect of glucagon-like peptide 2 pretreatment on growth performance and intestinal development of weaned piglets.

BackgroundGlucagon-like peptide 2 (GLP-2) is a potent epithelium-specific intestinal growth factor. The aim of this study was to demonstrate the prolonged effect of GLP-2 on the growth performance of weaned piglets. Forty piglets weaned at the age of 28 d with an average BW of 6.8 ± 0.4 kg were assigned to four treatments: (i) non-challenged control; (ii) LPS-challenged control; (iii) LPS + low GLP-2; and (iv) LPS + high GLP-2. Piglets in groups (i), (ii), and (iv) were s.c. injected with PBS supplemented with human [Gly2]GLP-21-34 at doses of 0, 2 and 10 nmol/kg BW per day for seven consecutive days. BW, gain:feed ratio (G:F), and plasma GLP-2 levels were determined on d 0, 7, and 14 after weaning. Piglets were challenged with i.p. administration of Escherichia coli lipopolysaccharide (LPS) at a dose of 100 μg/kg on d 14 to induce intestinal damage. Twenty-four hours later, intestinal tract samples were collected to assess intestinal morphology and quantify enzyme activity.ResultsPlasma GLP-2 levels decreased after weaning, but in the high GLP-2 group, plasma GLP-2 was maintained on d 7 and even increased to a level higher than the preweaning level on d 14 (P < 0.05). High GLP-2 treatment significantly increased the duodenal, jejunal and ileal weight, as well as the gross weight of the small intestine (SI), and the SI weight index (P < 0.05). LPS caused villous atrophy and disrupted intestinal morphology in the duodenum, jejunum and ileum. GLP-2 also significantly increased the villus height and the villus height/crypt depth ratio (VCR) of the duodenum, jejunum, and ileum (P < 0.05). Histological examination revealed that in GLP-2-treated groups, the integrity of the villus was maintained, and the villus was protected against LPS-induced damage. GLP-2 significantly increased the activity of alkaline phosphatase (AKP), γ-glutamyltranspeptidase (γ-GT), and pancreatic lipase in the duodenum and jejunum (P < 0.05). GLP-2 treatment also significantly increased the average daily gain (ADG) and G:F of piglets at 0 to 7, 7 to 14, as well as 0 to14 d (P < 0.05), resulting in a significant increase of final BW in high GLP-2 pigs (P = 0.016).ConclusionsExogenous GLP-2 improved the growth of weaned piglets and protected them against LPS-induced intestinal damage. These effects may be due to the ability of GLP-2 to promote the secretion of endogenous GLP-2 to stimulate the small intestinal development.

Monitoring γ-glutamyl transpeptidase activity and evaluating its inhibitors by a water-soluble near-infrared fluorescent probe

The first near-infrared fluorescent probe with excellent water-solubility for γ-glutamyl transpeptidase (GGT) has been developed by combining glutathione (GSH) as a recognition unit with a near-infrared hemicyanine fluorophore through an acrylyl linker. The probe exhibits a highly selective and sensitive fluorescent off-on response to GGT with a detection limit of 0.50U/L, and the response mechanism is based on the enzyme-catalyzed cleavage of the γ-glutamyl bond of GSH, followed by the spontaneous intramolecular cyclization and the release of the fluorophore. Notably, the probe has been used to image GGT in zebrafish and evaluate the inhibition ability of three common inhibitors of GGT both in vitro and in vivo, revealing that their inhibition efficiencies are acivicin >6-diazo-5-oxo-L-norleucine >L-serine-borate complex, and their corresponding IC50 values are 0.11±0.01mM, 0.34±0.04mM and 2.06±0.24mM, respectively. The proposed probe is simple, and may have great potential for screening GGT inhibitors.

Effects of Lavender Oil on Hair Growth-Relevant Enzyme Activity and Cytokine Expression in C57BL/6 Mice

The purpose of this study was to determine the effects of lavender oil (LO) on hair growth-relevant enzyme activity and cytokine expression in C57BL/6 mice. The experimental animals were divided into the following groups: i) normal (N: saline), ii) vehicle control (VC: jojoba oil), iii) positive control (PC: 3% minoxidil), iv) experimental 1 (E1: 3% LO), and v) experimental 2 (E2: 5% LO). Test compound solutions were topically applied to the backs of the mice (100 μL per application), once per day, 5 times a week, for 4 weeks. At week 4, the activities of γ-glutamyl transpeptidase (GT) in the E1 and E2 groups were 11% and 15% higher than the N group (p<0.05), respectively. The expression levels of keratinocyte growth factor (KGF) mRNA were significantly greater [10%; ( p<0.05)] in the E1 and E2 groups, compared to the N group. The expression of insulin-like growth factor (IGF)-1 mRNA also significantly increased by 21% and 22% (p<0.05) in the E1 and E2 groups, compared to the N group, respectively. Hepatocyte growth factor (HGF) mRNA expression levels were significantly higher in the E1 and E2 groups than the N group [14% and 17%; (p<0.05)], respectively. The expression of vascular endothelial growth factor (VEGF) mRNA increased by 22% (p<0.05) in both the E1 and E2 groups, when compared to the N group. The correlation coefficients among hair growth-relevant factors were significantly high each other (p<0.001). The correlation coefficient between hair growth area and KGF mRNA expression was 0.940. These results confirm that LO has hair growth-promoting effects as identified by hair growth-relevant enzyme activity and cytokine expression in C57BL/6 mice.

고지방식이와 저용량 스트렙토조토신으로 유도된 2형 당뇨병 흰쥐에서 녹차 폴리페놀과 비타민 C 병합 투여 효과

녹차 폴리페놀과 비타민 C가 2형 당뇨병에 미치는 영향을 관찰하기 위해 고지방식이와 저용량 스트렙토조토신을 이용하여 당뇨병이 유도된 흰쥐에 polyphenon 60(P60)과 sodium ascorbate(SA)를 16주간 투여한 후 공복 혈당, 혈장 인슐린, 혈청 중성지질, BUN 및 creatinine과 신장 GGT와 AP 활성도를 측정하였다. 당뇨병 흰쥐에 P60이나 SA투여로 인한 식이섭취량과 체중의 변화는 없었다. 공복 혈당은 당뇨 대조군(DC 군)에서 대조군(NC 군)의 5배로 증가하였으며, 당뇨+녹차 폴리페놀(DM+P60) 군은 14%, 당뇨+녹차 폴리페놀+비타민 C(DM+P60+SA) 군은 28%가 DC군보다 감소하였고, 당뇨+비타민 C(DM+SA) 군은 DC 군과 유의한 차이가 없었다. 혈장 인슐린 양은 각 실험군 사이에 유의한 차이가 없으며, 인슐린 저항성 지수 HOMA-IR은 DC 군에서 증가하였고 DM+P60+SA 군은 DC 군보다 감소하였다. 혈청 creatinine 양은 DC 군에서 증가하였고 DM+P60 군은 DC 군보다 17% 감소하였으며, DM+P60+SA 군은 DC 군보다 43% 감소하였다. 혈청 BUN 양은 DC 군에서 증가하였고 DM+P60+SA 군은 DC 군보다 41% 감소하였다. 신장 GGT와 AP 활성도는 DC 군에서 감소하였고 DM+P60+SA 군은 DC 군보다 감소가 적었으며, DM+SA군은 DC 군과 유의한 차이가 없었다. 이상의 실험 결과 2형당뇨병에서 녹차 폴리페놀과 비타민 C 복합 투여가 혈당, 인슐린 저항성 및 혈청 중성지질량 개선 및 신장 손상 방지효과가 녹차 폴리페놀이나 비타민 C를 단독으로 투여하였을 때보다 더 증가함을 확인할 수 있었다. 녹차 폴리페놀과 비타민 C의 혈당 상승 억제와 신장 손상 방지 효과에 대한 계속된 연구가 필요하며, 녹차 폴리페놀과 비타민 C는 당뇨병 환자에게 혈당을 조절하고 신장 손상을 방지하는 기능성식품 제재로서 개발 가능성이 있는 것으로 생각한다.

Effect of dietary cottonseed meal on growth performance, physiological response, and gossypol accumulation in pre-adult grass carp, Ctenopharyngodon idellus

Cottonseed meal (CM) was used at up to 36.95% content in the diet (replacing 60% of dietary fish meal protein) without any negative eff ects on growth performance of pre-adult grass carp (initial body weight, 761 g) under outdoor conditions. A culture trial was conducted in net cages installed in a large concrete pond. Seven isonitrogenous and isoenergetic diets containing a gradient of CM concentrations (0, 12.2%, 24.4%, 36.6%, 48.8%, 54.8%, and 61.0%) as replacement for dietary fish meal protein (0, 20%, 40%, 60%, 80%, 90%, and 100%) were formulated. Dietary non-resistant starch (from maize) was inverse to dietary CM. Growth performance and feed utilization of fish fed the diets containing CM replacing 0–40% fishmeal protein were not aff ected after the 6-week feeding trial. Accumulation of hepatopancreatic total gossypol in the hepatopancreas was significantly correlated with free gossypol content in the diets (HTG=88.6+1.5×DFG, R2=0.89, P<0.05). Intestinal α-amylase and γ-glutamyl transpeptidase activities rose along with increasing dietary CM level. The structure of the mid-intestinal tissues and the ultrastructure of the enterocyte microvilli were normal when dietary CM was <36.6% (60% protein replacement). Increasing dietary CM content increased serum alanine aminotransferase levels but decreased serum alkaline phosphatase, cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, and albumin (P<0.05).

The Ginkgo biloba Extract Reverses the Renal Effects of Titanium Dioxide Nanoparticles in Adult Male Rats.

The Ginkgo biloba extract (GbE) is a commercial product used as a nutraceutic herbal remedy in Europe and US. It contains 27% of the polyphenols isorhamnetin, kaempferol, and quercetin, as antioxidants. We used male adult Wistar rats (200–300 g), divided into four groups: control group (treated with 5.0 mg/kg of sodium chloride, intravenous), titanium dioxide nanoparticles (TiO2-NPs) group (5.0 mg/kg, intravenous), GbE group (10 mg/kg, intraperitoneal), and GbE + TiO2-NPs group (treated 24 h before with 10 mg/kg of GbE, intraperitoneal), followed, 24 h later, by 5.0 mg/kg of TiO2-NPs intravenously. The statistical analysis was performed using Student's t-test for grouped data with ANOVA posttest. The GbE protected renal cells against the effects of TiO2-NPs because it reversed the increased activity of γ-glutamyltranspeptidase and the enzymatic activity of dipeptidylaminopeptidase IV at all times tested (0–5, 5–24, 24–48, and 48–72 h). Also it reversed the glucosuria, hypernatriuria, and urine osmolarity at three times tested (5–24, 24–48, and 48–72). Thus, we conclude that GbE has a beneficial activity in the cytoplasmic membranes of brush border cells on the renal tubules, against the adverse effects that can be produced by some xenobiotics in this case the TiO2-NPs, in experimental rats.

Determination of enzyme activity in rabbit seminal plasma and its relationship with quality semen parameters

&lt;p&gt;The objective of this study was to determine rabbit seminal plasma enzyme activity. Furthermore, correlations between semen parameters and enzyme activity and male age were examined. The study was performed using 17 New Zealand White males from 5 to 9 mo old. Overall, 252 semen samples were collected from bucks from May to September. Semen characteristics were analysed and the seminal plasma was obtained by centrifugation. The activities of alanyl aminopeptidase (APN), aspartate transaminase (AST), alanine aminotransferase (ALT), γ-glutamyl transpeptidase (GGT), lactate dehydrogenase (LDH) and alkaline phosphatase (ALKP) in the seminal plasma fluid were measured. Significant differences between males were found in APN, GGT, LDH, ALKP and ALT activities (P&amp;lt;0.05). No significant differences between enzyme activity and male age were found. We also observed significant positive correlations between male age and sperm concentration (r=0.26), progressive motility (r=0.17) and amplitude of lateral head displacement (r=0.21), and negative ones between male age and average path velocity (r=–0.56), velocity of the sperm head along its actual curvilinear path (r=–0.61), straight line velocity (r=–0.50), linearity index (r=–0.13), and cytoplasmic droplet (r=–0.33). Furthermore, a significant negative correlation between APN activity and the status of the acrosome (r=–0.20) and significant positive correlations between APN activity and the sperm abnormalities (r=0.21), GGT activity and sperm concentration (r=0.34) and the status of the acrosome (r=0.31), and ALKP activity and sperm concentration were observed (r=0.41). In our study, APN and GGT seem to be the most predictive enzymes for rabbit semen quality.&lt;/p&gt;