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In vitro assay for HCV serine proteinase expressed in insect cells

World Journal of Gastroenterology 2003年第7期9卷 1629-1632页

作者： Li-Hua Hou Gui-Xin Du Rong-Bin Guan Yi-Gang Tong Hai-Tao Wang, department of applied Molecular Biology, Institute of microbiology and Epidemiology, Beijing 100071, China department of applied Molecular Biology Institute of Microbiology and Epidemiology Beijing 100071 China. houlihua@***

AIM: To produce the recombinant NS3 protease of hepatitis C virus with enzymatic activity in insect ***: The gene of HCV serine proteinase domain which encodes 181 amino acids was inserted into pFastBacHTc and the recombinant plasmid pFBCNS3N was transformed into DH10Bac competent cells for *** the recombinant bacmids had been determined to be correct by both blue-white colonies and PCR analysis, the isolated bacmid DNAs were transfected into Sf9 insect *** bacmids DNA was verified to replicate in insect cells and packaged into baculovirus particles via PCR and electronic microscopic analysis. The insect cells infected with recombinant baculovirus were determined by SDS-PAGE and Western-blot assays. The recombinant protein was soluted in N-lauryl sarcosine sodium (NLS) and purifed by metalchelated-affinity chromatography, then the antigenicity of recombinant protease was determined by enzyme-linked immunoabsorbant assay and its enzymatic activity was ***: The HCV NS3 protease domain was expressed in insect cells at high level and it was partially solved in *** 0.2 mg recombinant serine proteinase domain with high purity was obtained by metal-chelated-affinity chromatography from 5×107 cells, and both antigenicity and specificity of the protein were evaluated to be high when used as antigen to detect hepatitis C patients′ sera in indirect ELISA format. In vitro cleavage assay corroborated its enzymatic ***: The recombinant HCV NS3 proteinase expressed by insect cells is a membrane-binding protein with good antigenicity and enzymatic activity.

关键词：丙型肝炎病毒丝氨酸蛋白酶基因重组聚合酶链反应

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Assessment of probiotic and antifungal activity of Lactiplantibacillus plantarum MYSAGT3 isolated from locally available herbal juice against mycotoxigenic Aspergillus species

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Food Bioscience 2022年第6期50卷 1549-1560页

作者： G.Adithi Rakesh Somashekaraiah S.Divyashree B.Shruthi M.Y.Sreenivasa applied Mycology Lab Department of Studies in MicrobiologyUniversity of MysoreManasagangotriMysuruIndia

This study investigated the antifungal activities of probiotic bacteria isolated from traditional herbal juice, which was a blend of Tulsi, Mint, Bael and Indian borage leaves. About 20 isolates were obtained from the herbal juice sample;these isolates were initially screened against fungal pathogens such as Aspergillus parasiticus , Aspergillus flavus , Aspergillus ochraceus , Aspergillus carbonarius and Aspergillus fumigatus . Of the 20 isolates obtained, MYSAGT3 and MYSAGT4 exhibited potential antifungal activity against the tested fungal pathogens. Moreover, MYSAGT3 and MYSAGT4 was found to survive acidic pH and bile conditions, but were susceptible to tested antibiotics. Both the isolates exhibited 40–60% of autoaggregation ability, >90% of cell surface hydrophobicity and tolerance to 0.4–0.6% phenol. The 16S rRNA sequencing results identified the isolates MYSAGT3 and MYSAGT4 as Lactiplantibacillus plantarum. The MYSAGT3 exhibited potent antagonistic and probiotic activity compared to MYSAGT4;hence MYSAGT3 was further used for the control of Aspergillus ochraceus. Initial antifungal activity of MYSAGT3 inhibited about 76.67% growth of Aspergillus ochraceus comparatively higher than other Aspergillus sp. tested. Co-inoculation and microdilution assay revealed the high inhibitory activity of Lactiplantibacillus plantarum MYSAGT3. Further, cell free supernatant (CFS) of Lactiplantibacillus plantarum MYSAGT3 reduced the growth of fungal hyphae and conidial germination of Aspergillus ochraceus in conidial germination assay and bio preservative activity on peanuts. The CFS characterization and Liquid chromatography-mass spectroscopy (LCMS) results showed that, the antifungal activity was due to different organic acids present in the CFS of MYSAGT3. MYSAGT3 and its CFS also reduced the growth of Aspergillus ochraceus in Poultry feeds and about 21.80% of ochratoxin A (OTA) degradation was attained after 24 h of incubation with MYSAGT3. The isolated Lactic acid bacteria

关键词： Aspergillus ochraceus Lactiplantibacillus plantarum Cell free supernatant Antifungal activity Ochratoxin A

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Prevalence and co-infections of schistosomiasis/hepatitis B and C viruses among school children in an endemic areas in Taiz,Yemen

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Asian Pacific Journal of Tropical Medicine 2011年第5期4卷 404-408页

作者： Adam H Al-Shamiri Mohammed A Al-Taj Ameera S Ahmed department of microbiology Faculty of Applied ScienceTaiz UniversityYemen department of Parasitology and Immunology Faculty of Applied ScienceTaiz UniversityYemen

Objective:To determine the disease prevalence and its relationship with hepatitis B and C viruses among school children in five endemic areas by ***:During June 2007 and March 2009,1 484 school children aged between 5-16 years participated in the current study from 32 basic schools in five districts(Al-Dhabab,Hedran,Warazan,Al-Barh and Al-Shmaytin) in Taiz Governorate,Republic of *** of school children who participated in the study;1 406 stool samples,1 484 urine samples and 214 blood samples were collected and ***:Schistosoma mansoni(***) was found in all the studied areas except ***,Schistosoma haematobium(***) was recorded only in Al-Shmaytin and *** *** and *** were observed in Al-Shmaytin *** overall prevalence was 20.76%for *** and 7.41%for ***. The prevalence rate of infection among males was higher than females,shwing no significant *** of light,moderate and heavy infections in the case of *** were 41.78%, 25.34%and 32.87%***,for *** it was 50.90%for light infection and 49.09%for heavy *** to the prevalence of viral hepatitis among infected school children with schistosomiasis,it could be noticed that hepatitis B virus was higher than the prevalence of hepatitis C ***,the presence of HBsAg and anti-HCV was not associated with Schistosoma ***:Schistosomiasis infection is an important public health problem in Taiz Governorate,Republic of *** was a correlation between S. haematobium and hepatitis B,but no association between *** infections and hepatitis B and C viruses.

关键词： Schistosomosis Intensity Hepatitis B and C virus School children Yemen

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Lignin Biosynthesis Studies in Plant Tissue Cultures

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Journal of Integrative Plant Biology 2010年第2期52卷 176-185页

作者： Anna Krknen Sanna Koutaniemi department of applied Biology 00014 University of HelsinkiFinland MTT Agrifood Research Finland 92400 RuukkiFinland department of applied Chemistry and microbiology 00014 University of HelsinkiFinland

Lignin, a phenolic polymer abundant in cell walls of certain cell types, has given challenges to scientists studying its structure or biosynthesis. In plants lignified tissues are distributed between other, non-lignified tissues, Characterization of native lignin in the cell wall has been difficult due to the highly cross-linked nature of the wall components. Model systems, like plant tissue cultures with tracheary element differentiation or extracellular lignin formation, have provided useful information related to lignin structure and several aspects of lignin formation. For example, many enzyme activities in the phenylpropanoid pathway have been first identified in tissue cultures. This review focuses on studies where the use of plant tissue cultures has been advantageous in structural and biosynthesis studies of lignin, and discusses the validity of tissue cultures as models for lignin biosynthesis.

关键词： cell Lignin Biosynthesis Studies in Plant Tissue Cultures

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<i>In Vivo</i>Study of Healing Effects of Sida acuta Leaf Extracts on <i>Helicobacter pylori</i>Induced Ulceration in Mice

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Journal of Biosciences and Medicines 2020年第9期8卷 1-14页

作者： Chito Clare Ekwealor Chidinma Chinwendu Okorie Chinwe Clarice Ukoha Anthonia Nkiruka Mba department of applied microbiology & brewing Nnamdi Azikiwe University Awka Nigeria department of microbiology Chukwuemeka Odimegwu Ojukwu University Uli Nigeria

Helicobacter pylori is a human pathogen that is widely distributed and known to cause peptic ulcer, gastritis and gastric cancer. Treatment is usually by combination of acid inhibitory therapy and antibiotics because of drug resistance problems. Studies now focus on the use of medicinal plants for treatment of H. pylori induced ulcers because they are inexpensive and have limited side effects. This study was undertaken to determine the effects of cold water and ethanol extracts of Sida acuta leaves on H. pylori induced ulcer in mice. Oral administration with the aid of a feeding tube of 0.5 ml of 107 CFU/ml of H. pylori (Accession number LT799736) in phosphate buffer saline (pH 6.8) was used to induce ulcer in mice. Effects of various concentrations of cold water and ethanol extracts of S. acuta leaves on the ulcer parameters and histology examinations of stomach tissues of mice were investigated after 2 weeks of administration of extracts into the mice. Mice treated with clarithromycin served as positive control while mice administered with H. pylori but not treated served as negative control. One-way analysis of variance (ANOVA) was used for data analysis and results were considered significant if p 6 CFU/ml to 3.5 × 106 CFU/ml and 2.6 × 106 CFU/ml for cold water and ethanol extracts respectively. Reduction in ulcer severity (2.00 ± 0.10 to 0.080 ± 0.05), ulcer index (8.50 ± 1.10 to 4.26 ± 0.03) and gastric volume (1.32 ± 0.12 to 0.54 ± 0.13) were observed. Histology of stomach tissues of mice treated with extracts revealed clear granulation indicating tissue repair and wound healing. S. acuta extracts were observed to enhance ulcer healing in a mice model.

关键词： Helicobacter pylori Sida acuta Leaves Peptic Ulcer Ethanol Extract

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Impact of Helicobacter pylori virulence markers on clinical outcomes in adult populations

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World Journal of Gastroenterology 2023年第1期29卷 190-199页

作者： Halim Roshrosh Hanan Rohana Maya Azrad Tamar Leshem Segula Masaphy Avi Peretz applied Mycology and microbiology MigalKiryat Shemona 1101202Israel department of microbiology Padeh Poriya Medical CenterTiberias 111508Israel

BACKGROUND In recent years,associations between specific virulence markers of Helicobacter pylori(***)and gastrointestinal disorders have been *** To investigate the presence of virulence factors including vacuolating cytotoxin A genotypes(s1m1,s1m2,s2m1,and s2m2),cytotoxin-associated gene A(CagA),and urease activity in *** strains isolated from Arab and Jewish populations in northern Israel and to assess associations between these factors and patients’demographics and clinical *** Patients(n=108)who underwent gastroscopy at the Baruch Padeh Medical Center,Poriya due to symptomatic gastroduodenal pathologies as part of *** diagnosis were enrolled in the *** biopsy specimens were collected from the antrum of the *** condition was assessed by clinical pathology *** were isolated on modified BD Helicobacter Agar(BD Diagnostics,Sparks,MD,United States).Bacterial DNA was extracted,and PCR was performed to detect CagA and vacuolating cytotoxin A *** activity was assessed using a rapid urease *** A significant correlation was found between disease severity and patient ethnicity(P=0.002).A significant correlation was found between CagA presence and the s1m1 genotype(P=0.02),which is considered the most virulent ***,a higher level of urease activity was associated with isolates originating from the Jewish ***,higher urease activity levels were measured among CagA-/s1m1 and CagA-/s2m2 *** Our study highlights the importance of incorporating molecular methods for detection of virulence markers of *** in order to tailor optimal treatments for each *** investigation should be performed regarding associations between *** virulence factors and ethnicity.

关键词： Helicobacter pylori Virulence factors Vacuolating cytotoxin A Cytotoxin-associated gene A Urease activity

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Purification and molecular weight distribution of a key exopolysaccharide component of Bacillus megaterium TF10

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Journal of Environmental Sciences 2018年第1期30卷 9-15页

作者： Lingling Wang Longfei Wang Qingshan Shi Hanqing Yu State Key Laboratory of applied microbiology Southern China Guangdong Provincial Key Laboratory of Microbial Culture Collection and ApplicationGuangdong Institute of Microbiology department of Chemistry University of Science & Technology of China

Extracellular polymeric substances（EPS） are organic metabolic compounds excreted by microorganisms. They largely impact microbial aggregate structures and *** polysaccharides（EP） in EPS are responsible for the formation of microbial aggregates. In this work, we successfully separated and characterized EP from EPS of the bacterium Bacillus megaterium TF10. Extraction of EP from EPS was optimized using Sevag＇s reagent. Chemical characteristics, functional groups, and molecular weight（MW） distribution of EP were compared with the harvested EPS and soluble microbial products（SMP）. We found that the polymers of lower MW and free proteins were successfully removed by Sevag＇s reagent. The higher MW components of EPS were predominantly polysaccharides,while the polymers of lower MW tended to secrete to the supernatant and were described as SMP. A part of the proteins in the EP was polysaccharide-bonded. Our results can be further used in elucidating the complex flocculation mechanisms in which EP play a major role.

关键词： Extracellular polysaccharides(EP) Extracellular polymeric substances(EPS) Soluble microbial products(SMP) Molecular weight(MW) distribution

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Differential degradation of crude oil (Bonny Light) by four Pseudomonas strains

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Journal of Environmental Sciences 2009年第2期21卷 243-248页

作者： Oluwafemi S.Obayori Sunday A.Adebusoye Adams O.Adewale Ganiyu O.Oyetibo Odunola O.Oluyemi Rashid A.Amokun Matthew O.Ilori department of microbiology Lagos State University department of Botany and microbiology University of Lagos

Four hydrocarbon degraders isolated from enriched oil- and asphalt-contaminated soils in Lagos, Nigeria, were tested for their petroleum degradation potentials. All the isolates were identified as species of Pseudomonas. Pseudomonas putida P 11 demonstrated a strong ability to degrade kerosene, gasoline, diesel, engine oil and crude oil while P. aeruginosa BB3 exhibited fair degradative ability on crude oil, gasoline, engine oil, anthracene and pyrene but weak on kerosene, diesel and dibenzothiophene. Pseudomonas putida WL2 and P. aeruginosa MVL1 grew on crude oil and all its cuts tested with the latter possessing similar polycyclic aromatic potentials as P11. All the strains grew logarithmically with 1-2 orders of magnitude and with generation time ranging significantly between 3.07 and 8.55 d at 0.05 level of confidence. Strains WL2 and MVL1 utilized the oil substrate best with more than 70% in 6 d experimental period, whereas the same feat was achieved by P11 in 12 d period. BB3 on the other hand degraded only 46% within 6 d. Interestingly, data obtained from gas chromatographic analysis of oil recovered from the culture fluids of MVL1 confirmed near-disappearance of major peaks （including aliphatics and aromatics） in the hydrocarbon mixture.

关键词： biodegradation crude oil hydrocarbons pollution Pseudomonas strains

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Comprehensive characterization of indoor airborne bacterial profile

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Journal of Environmental Sciences 2009年第8期21卷 1148-1152页

作者： P. L. Chan P. H. F. Yu Y. W. Cheng C. Y. Chan P. K. Wong department of Biology The Chinese University of Hong Kong department of applied Biology and Chemical Technology The Hong Kong Polytechnic University department of microbiology The Chinese University of Hong Kong

This is the first detailed characterization of the airborne bacterial profiles in indoor environments. Two restaurants were selected for this study. Fifteen genera of bacteria were isolated from each restaurant and identified by three different bacterial identification systems including MIDI, Biolog and Riboprinter. The dominant bacteria of both restaurants were Gram-positive bacteria in which Micrococcus and Bacillus species were the most abundant. Most bacteria identified were representative species of skin and respiratory tract of human, and soil. Although the bacterial levels in these two restaurants were below the limit of the Hong Kong Indoor Air Quality Objective （HKIAQO） Level 1 standard （i.e., 〈 500 cfu/m^3）, the majority of these bacteria were opportunistic pathogens. These results suggested that the identity of airborne bacteria should also be included in the IAQ to ensure there is a safety guideline for the public.

关键词： airborne bacterial profile bacterial identification indoor air quality

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Effects of shifting cultivation on biological and biochemical characteristics of soil microorganisms in Khagrachari hill district, Bangladesh

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Journal of Forestry Research 2014年第3期25卷 689-694页

作者： Sohag Miah S.M. Sirajul Haque Wahida Sumi Mohammad Mosharraf Hossain Institute of Forestry and Environmental Sciences University of Chittagong department of microbiology University of Chittagong department of applied Physics Graduate School of Engineering Osaka University

We collected soil samples from two representative sites at Aatmile of Khagarachari hill district in Chittagong Hill Tracts. One of the sites was under shifting cultivation and the other an adjacent 13-year old teak plantation. Both sites were in the same physiographic condition and same aspect with parable soil type, which enabled us to measure the effects of shifting cultivation on soil micro-flora. We studied soil phys-ico-chemical properties and the biochemical and biological properties of soil microbes. Moisture and organic matter content as well as fungi and bacterial populations, both in surface and subsurface soils, were signifi-cantly （p≤0.001） lower in shifting cultivated soils compared to soils not under shifting cultivation, i.e. the teak plantation site. The most abundant bacteria in surface （0-10 cm） and sub-surface （10-20 cm） soils under shifting cultivation were Pseudomonas diminuta and Shigella, respec-tively, while in corresponding soil layers of teak plantation, predominant microbes were Bacillus firmus （0-10 cm） and Xanthomonas （10-20 cm）. The microbial population differences cannot be explained by soil texture differences because of the textural similarity in soils from the two sites but could be related to the significantly lower moisture and organic mat-ter contents in soils under shifting cultivation.

关键词： shifting cultivation soil biological properties soil biochemical properties soil microflora Chittagong Hill Tracts

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