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Sensitive detection of alkaline phosphatase based on terminal deoxynucleotidyl transferase and endonuclease Ⅳ-assisted exponential signal amplification

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Journal of Pharmaceutical Analysis 2022年第4期12卷 692-697页

作者： Weicong Ye Longjie Li Zishan Feng Bocheng Tu Zhe Hu Xianjin Xiao Tongbo Wu Department of Pharmaceutical Analysis School of PharmacyTongji Medical CollegeHuazhong University of Science and TechnologyWuhan430030China Department of Biopharmaceutics School of Life Science and TechnologyWuhan Polytechnic UniversityWuhan430023China Department of Translational Medicine Institute of Reproductive HealthTongji Medical CollegeHuazhong University of Science and TechnologyWuhan430030China

Alkaline phosphatase(ALP)is widely expressed in human *** plays an important role in the dephosphorylation of proteins and nucleic ***,quantitative analysis of ALP plays a vital role in disease diagnosis and the development of biological detection *** deoxynucleotidyl transferase(TdT)catalyzes continuous polymerization of deoxynucleotide triphosphates at the 30-OH end of single-stranded DNA in the absence of a *** this study,we developed a highly sensitive and selective method based on TdT and endonuclease Ⅳ(Endo Ⅳ)to quantify ALP *** ALP hydrolyzes the 30-PO_(4) end of the substrate and generates 30-OH,TdT can effectively elongate the 30-OH end with deoxynucleotide adenine triphosphate(dATP)and produce a poly A tail,which can be detected by the poly T *** Ⅳ digests the AP site in poly T probes to generate a fluorescent signal and a new 30-OH end,leading to the generation of exponential fluorescence signal *** substrate for TdT elongation was optimized,and a limit of detection of 4.3×10^(-3) U/L was achieved for ALP by the optimized substrate *** method can also detect ALP in the cell lysate of a single *** work has potential applications in disease diagnosis and biomedical detection.

关键词： Alkaline phosphatase Terminal deoxynucleotidyl transferase EndonucleaseⅣ Exponential amplification

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Association of serum gamma-glutamyl transferase with treatment outcome in chronic hepatitis B patients

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World Journal of Gastroenterology 2015年第34期21卷 9957-9965页

作者： Rui Huang Chen-Chen Yang Yong Liu Juan Xia Ran Su Ya-Li Xiong Gui-Yang Wang Zhen-Hua Sun Xiao-Min Yan Shan Lu Chao Wu Department of Infectious Diseases Nanjing Drum Tower HospitalNanjing University Medical School Department of Infectious Diseases Qidong People’s Hospital Department of Laboratory Medicine Nanjing Drum Tower HospitalNanjing University Medical School Department of Medicine University of Massachusetts Medical School

AIM:To investigate the association of serum gammaglutamyl transferase(GGT) levels with chronic hepatitis B infection and hepatitis B e antigen(HBe Ag) ***:A retrospective study was performed on clinical data collected from patients who had been positive for hepatitis B surface antigen for > 6 mo and who were antiviral-treatment na?ve(n = 215) attending the Hepatitis Clinic at Nanjing Drum Tower Hospital between August 2010 and December 2013. Healthy individuals without liver disease(n = 83) were included as controls. Patients were categorized into four groups based on disease status as recommended by the European Association for the Study of the Liver:immune tolerance(IT; n = 47),HBe Ag-positive hepatitis(EPH; n = 93),HBe Ag-negative hepatitis(ENH;n = 20),and inactive carrier(IC; n = 55). Prediction of complete response(CR) based on serum GGT was also examined in EPH patients(n = 33) treated for 48 wk with nucleos(t)ide analogue(NA) therapy,including lamivudine plus adefovir combination therapy(n = 20) or entecavir monotherapy(n = 13). CR was defined as a serum hepatitis B virus DNA level < 500 copies/m L and HBe Ag seroconversion by 48 wk of treatment. RESULTS:Serum GGT levels were significantly increased in EPH and ENH patients relative to the IT,IC,and healthy control groups(P < 0.01 for all). However,no significant difference in serum GGT levels was found between the EPH and ENH groups. Baseline serum GGT levels were significantly higher in patients who achieved CR(7/33; 21.2%) compared to patients in the non-CR group(26/33; 78.8%; P = 0.011). In addition,the decline in serum GGT was greater in CR patients compared to non-CR patients after 24 wk and 48 wk of treatment(P = 0.012 and P = 0.008,respectively). The receiver operating characteristic curve yielded a sensitivity of 85.71% and a specificity of 61.54% at a threshold value of 0.89 times the upper limit of normal for baseline serum GGT in the prediction of CR following NA therapy. CONCLUS

关键词： Gamma-glutamyl transferase Hepatitis B e antigen H

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Effect of Lactobacillus brevisSBC8803 on Gamma-Glutamyl transferase in Japanese Habitual Drinkers: A Double-Blind, Placebo-Controlled Study

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Food and Nutrition Sciences 2012年第5期3卷 678-684页

作者： Yoshihisa Wakita Hajime Kanda Chikako Shimizu Yasukazu Nakakita Hirotaka Kaneda Shuichi Segawa Moeko Ozaki Tatsuro Shigyo Takaaki Ohtake Mikihiro Fujiya Yutaka Kohgo Corporate Planning Department Sapporo Holdings Ltd. Tokyo Japan Division of Gastroenterology and Hematology/Oncology Department of Medicine Asahikawa Medi- cal University Hokkaido Japan Frontier Laboratories of Value Creation Sapporo Breweries Ltd. Shizuoka Japan

A randomized, double-blind, placebo-controlled clinical trial in Japanese habitual drinkers was conducted to evaluate the efficacy of Lactobacillus brevis SBC8803 to alleviate adverse effect of alcohol. Subjects who drank habitually and had moderately higher levels of gamma-glutamyl transferase (GGT) (50 - 100 IU/L) were enrolled. The levels of transaminases in these subjects were almost within normal levels (aspartate transaminase (AST) <30 IU/L and alanine transaminase (ALT) <40 IU/L). Either the capsules containing placebo (n = 23) or 130 mg (4.0 × 1010 colony-forming units) of live L. brevis SBC8803 (n = 22) per day were administered for the continuous eight weeks (56 days). During the period, the subjects both in test group and placebo groups have kept each drinking behavior as usual. Regarding lipid metabolism, triacylglycerol (TG) levels in the male test group significantly decreased at week 4 as compared with week 0. Biomarkers of hepatocytes-damage;AST and ALT levels showed no significant differences between the pla- cebo and test groups at both weeks 4 and 8. Oxidative stress marker;GGT at weeks 4 was significantly lower in the test group than that in the placebo group (p = 0.017), but not at weeks 8. However, taking a reduced rate of GGT at weeks 8 comparing with that at week 0, that in the test group showed larger value comparing with that in the placebo group. These data about TG and GGT suggest that, although efficacy of L. brevis SBC8803 is limited in this study, intake of the probiotic may alleviate alcoholic influence in lipid metabolism and oxidative stress.

关键词： Lactobacillus brevis Gamma-Glutamyl transferase Alcoholic Related Disorder Double-Blind Placebo-Controlled Study

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Glutathione transferase-P1-1 binding with naturally occurring ligands: assessment by docking simulations

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Journal of Biophysical Chemistry 2011年第4期2卷 401-407页

作者： Anupam J. Das Sreeda Chalil Poonam Nigam Pamela Magee Omar Janneh Richard Owusu-Apenten School of Biomedical Sciences University of Ulster Department of Bimolecular and Sport Sciences Coventry University

Glutathione transferase-P1-1 (hGSTP1-1), which is associated with acquired drug resistance in some tumour cells, requires two identical subunits for full activity. Naturally occurring inhibitors for GSTP1-1 quaternary structure could be interesting therapeutic agents. The aim of this study was to investigate potential binding sites for hGSTP1-1 interaction with ligands many of which occur naturally. Simulations were performed with commercial docking software and with GST monomer or dimer as template. Docking results using hGSTP1-1 dimer showed one binding site for most of the ligands tested. Lycopene, glutathione, ellagic acid, ethacrynic acid, quercetin, caffeic acid, ferulic acid, porphyrin, curcumin, cinnamic acid, and also α-tocopherol bound at the enzyme dimer subunit-subunit interface. In contrast, investigations using hGSTP1-1 monomer revealed three additional sites for ligand binding. In conclusion, the docking simulations suggest that the enzyme subunit interface may be important for hGSTP1-1 interactions with ligands. These findings may provide valuable insights for further research to identify naturally occurring therapeutic agents.

关键词： Glutathione transferase Docking Vitamins Phytochemicals

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Description of a Putative Oligosaccharyl:S-Layer Protein transferase from the Tyrosine O-Glycosylation System of Paenibacillus alveiCCM 2051T

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Advances in Microbiology 2012年第4期2卷 537-546页

作者： Robin Ristl Bettina Janesch Julia Anzengruber Agnes Forsthuber Johanna Blaha Paul Messner Christina Schaffer Department of NanoBiotechnology NanoGlycobiology Unit Universitat fur Bodenkultur Wien Wien Austria Department of NanoBiotechnology NanoGlycobiology Unit Universitat für Bodenkultur Wien Wien Austria

Surface (S)-layer proteins are model systems for studying protein glycosylation in bacteria and simultaneously hold promises for the design of novel, glyco-functionalized modules for nanobiotechnology due to their 2D self-assembly capability. Understanding the mechanism governing S-layer glycan biosynthesis in the Gram-positive bacterium Paenibacillus alvei CCM 2051T is necessary for the tailored glyco-functionalization of its S-layer. Here, the putative oligosaccharyl:S-layer protein transferase WsfB from the P. alvei S-layer glycosylation gene locus is characterized. The enzyme is proposed to catalyze the final step of the glycosylation pathway, transferring the elongated S-layer glycan onto distinct tyrosine O-glycosylation sites. Genetic knock-out of WsfB is shown to abolish glycosylation of the S-layer protein SpaA but not that of other glycoproteins present in P. alvei CCM 2051T, confining its role to the S-layer glycosylation pathway. A transmembrane topology model of the 781-amino acid WsfB protein is inferred from activity measurements of green fluorescent protein and phosphatase A fused to defined truncations of WsfB. This model shows an overall number of 13 membrane spanning helices with the Wzy_C domain characteristic of O-oligosaccharyl:protein transferases (O-OTases) located in a central extra-cytoplasmic loop, which both compares well to the topology of OTases from Gram-negative bacteria. Mutations in the Wzy C motif resulted in loss of WsfB function evidenced in reconstitution experiments in P. alvei ΔWsfB cells. Attempts to use WsfB for transferring heterologous oligosaccharides to its native S-layer target protein in Escherichia coli CWG702 and Salmonella enterica SL3749, which should provide lipid-linked oligosaccharide substrates mimicking to some extent those of the natural host, were not successful, possibly due to the stringent function of WsfB. Concluding, WsfB has all features of a bacterial O-OTase, making it the most probable candidate for

关键词： Bacterial Glycosylation S-Layer Oligosaccharyl transferase Tyrosine-O-Glycosylation Trans-Membrane Topology

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Repeat Whole Blood Donation Correlates Significantly with Reductions in BMI and Lipid Profiles and Increased Gamma Glutamic transferase (GGT) Activity among Nigerian Blood Donors

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Open Journal of Blood Diseases 2012年第4期2卷 90-94页

作者： Teddy C. Adias Anastecia Chika Igwilo Zaccheaus Awortu Jeremiah College of Health Technology Otuogidi Ogbia Nigeria Department of Haematology and Blood Transfusion University of Calabar Teaching Hospital Calabar Nigeria Haematology and Blood Transfusion Science Unit Department of Medical Laboratory Science College of Health Sciences Niger Delta University Wilberforce Island Nigeria

Background: The effect of repeated blood donation on some biochemical values of healthy adult male blood donors in Calabar, Nigeria was studied. Methods: One hundred and fifty three (153) healthy repeat blood donors and 90 first time blood donors constituted the study population. Samples were analyzed using colorimetric procedures. Results: The BMI values of 24.4 ± 2.4 kg/ m2 in the first time donors was significantly higher than the 21.7 ± 1.7 kg/m2 obtained in repeat donors (P 2 = 0.7934, P Conclusion: A reduction in the values of some lipid profiles and high GGT activity is associated with repeated blood donations in this study population. Repeated blood donation may play a significant role in reducing the incidence of heart disease.

关键词： Blood Donation Gamma Glutamic transferase Body Mass Index Lipid Profiles

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Association of adverse effects with monoamine oxidase type B inhibitor and catechol-o-methyl transferase inhibitor combination therapy in Parkinson’s disease patients

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Advances in Parkinson's Disease 2012年第1期1卷 5-10页

作者： Rui Zhang Danielle C. Spengler Marie-Helene Saint-Hilaire Anna D. Hohler Department of Neurology Boston University School of Medicine Boston USA Department of Neurology Boston University School of Medicine Boston USA

Currently, levodopa is the most effective and commonly used medication to control motor symptoms in Parkinson’s disease (PD). However, its long-term use is associated with adverse effects (AEs). Combination therapy of a monoamine oxidase type B inhibitor (MAOBI) with levodopa or a catechol-O-methyl transferase inhibitor (COMTI) with levodopa provides benefits to PD patients. Direct comparison of efficacy and side effect profiles is complex. The aim of this study is to investigate the different AE profiles of MAOBI and COMTI combination therapies. Data used to analyze the AEs of different PD medications were retrieved from “The Boston University Medical Center’s Parkinson’s Disease and Movement Disorder Database”. Ten categories of AEs were compared between patients receiving MAOBI and COMTI combination treatment. In total, 87 subjects were included in the analysis. Out of ten AEs, the presence of dementia was signifi- cantly different between the MAOBI and COMTI groups with an OR of 6.9 (COMTI vs MAOBI, 95% CI 1.3 - 37.0). Motor fluctuations were also found to be differently distributed in the two medication groups with an OR of 3.1 (COMTI vs MAOBI, 95% CI 1.0 - 9.8). In this retrospective database analysis of patients treated with combination treatment for PD, combination therapy of a COMTI with levodopa was more likely to be associated with dementia and motor fluctuations than a MAOBI with levodopa.

关键词： Catechol-O-Methyl transferase Inhibitors Dementia Monoamine Oxidase Type B Inhibitors Motor Fluctuations Parkinson’s Disease

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Challenges in diagnosing mesenteric ischemia

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World Journal of Gastroenterology 2013年第9期19卷 1338-1341页

作者： Teun C van den Heijkant Bart AC Aerts Joep A Teijink Wim A Buurman Misha DP Luyer Department of Surgery Catharina Hospital Eindhoven 5623 EJ Eindhoven The Netherlands Department of Surgery Amphia Hospital 4818 CK Breda The Netherlands Institute Nutrim Maastricht University 6200 MD Maastricht The Netherlands

Early identification of acute mesenteric ischemia (AMI) is challenging. The wide variability in clinical presentation challenges providers to make an early accurate diagnosis. Despite major diagnostic and treatment advances over the past decades, mortality remains high. Arterial embolus and superior mesenteric artery thrombosis are common causes of AMI. Non-occlusive causes are less common, but vasculitis may be important, especially in younger people. Because of the unclear clinical presentation and non-specific laboratory findings, low clinical suspicion may lead to loss of valuable time. During this diagnostic delay, progression of ischemia to transmural bowel infarction with peritonitis and septicemia may further worsen patient outcomes. Several diagnostic modalities are used to assess possible AMI. Multi-detector row computed tomographic angiography is the current gold standard. Although computed tomographic angiography leads to an accurate diagnosis in many cases, early detection is a persistent problem. Because early diagnosis is vital to commence treatment, new diagnostic strategies are needed. A non-invasive simple biochemical test would be ideal to increase clinical suspicion of AMI and would improve patient selection for radiographic evaluation. Thus, AMI could be diagnosed earlier with follow-up computed tomographic angiography or high spatial magnetic resonance imaging. Experimental in vitro and in vivo studies show promise for alpha glutathione S transferase and intestinal fatty acid binding protein as markers for AMI. Future research must confirm the clinical utility of these biochemical markers in the diagnosis of mesenteric ischemia.

关键词： Acute mesenteric ischemia Diagnosis Biological markers Intestinal fatty acid binding protein Alpha-glutathione S transferase

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Simple and effective detection method for gene expression product of transgenic plant

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Chinese Science Bulletin 1999年第10期44卷 912-916页

作者： Jiaxi Xu Mi Ma Yuan Ma Zhongping Lin Peking Univ Coll Chem & Mol Engn Beijing 100871 Peoples R China Chinese Acad Sci Inst Bot Beijing 100093 Peoples R China Tsing Hua Univ Dept Chem Bioorgan Phosphorus Chem Lab Beijing 100864 Peoples R China

It is a crucial problem in gene engineering whether an exogenous gene could be correctly expressed in cells of transgenic animal or plant, and how the expression products could be detected quantitatively in translational level. It is very difficult to analyze some gene expression products, such as isopentenyl transferase (ipt), in transgenic investigation because they are trace in organisms. A convenient method is described to determine trace content gene expression product which is hard to purify. The method includes predicting and synthesizing the antigenic peptide according to the cDNA sequence, coupling the synthetic antigenic peptide with carrier protein, raising specific antibodies against the synthetic antigen, and detecting the gene expression product by ELISA and Western blot.

关键词： transgenic plant isopentenyl transferase antigenic peptide detective method.

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Challenges in diagnosing adhesive small bowel obstruction

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World Journal of Gastroenterology 2013年第43期19卷 7489-7493页

作者： Thijs R van Oudheusden Bart AC Aerts Ignace HJT de Hingh Misha DP Luyer Department of Surgery Catharina Hospital5623 EJEindhovenThe Netherlands Department of Surgery Amphia Hospital4818 CKBredaThe Netherlands

Adhesive small bowel obstruction(ASBO)is the most frequently encountered surgical disorder of the small *** to 80%of ASBO cases resolve spontaneously and do not require invasive *** is important to identify such patients that will benefit from conservative treatment in order to prevent unnecessarily exposing them to the risks associated with surgical intervention,such as morbidity and further adhesion *** the remaining ASBO patients,timely surgical intervention is necessary to prevent small bowel strangulation,which may cause intestinal ischemia and bowel *** early identification of these patients is key to decreasing ASBO-related morbidity and mortality,the non-specific signs and laboratory findings upon clinic presentation limit timely diagnosis and implementation of appropriate clinical *** the clinical presentation findings with those from other diagnostic imaging modalities,such as abdominal X-ray,computed tomography-scan and water-soluble contrast studies,will improve diagnosis of ASBO and help clinicians to better evaluate the potential of conservative management as a safe strategy for a particular ***,patients who present with moderate findings by all these approaches continue to represent a challenge.A new diagnostic strategy is urgently needed to further improve our ability to identify early signs of strangulated bowel,and this diagnostic modality should be able to indicate when surgical management is required.A number of potential serum markers have been proposed for this purpose,including intestinal fatty acid binding protein andα-glutathione S ***-going research is attempting to clearly define their diagnostic utility and to optimize their potential role in determining which patients should be managed surgically.

关键词： Adhesive small bowel obstruction Diagnosis Clinical management Biological markers Intestinal fatty acid binding protein α-glutathione S transferase

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