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New GATEWAY vectors for High Throughput Analyses of Protein-Protein Interactions by Bimolecular Fluorescence Complementation

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molecular Plant 2009年第5期2卷 1051-1058页

作者： Christian Gehl Rainer Waadt jorg Kudla Ralf-R. mendel Robert Hansch Institut for Pflanzenbiologie Technische Universitat Braunschweig Humboldtstrasse 1 D-38106 Braunschweig Germany molekulare Entwicklungsbiologie der Pflanzen Institut for Botanik und Botanischer Garten Universitat MOnster Schlossplatz 4 D-48149 MOnster Germany

Complex protein interaction networks constitute plant metabolic and signaling systems. Bimolecular fluorescence complementation （BiFC） is a suitable technique to investigate the formation of protein complexes and the localization of protein-protein interactions in planta. However, the generation of large plasmid collections to facilitate the exploration of complex interaction networks is often limited by the need for conventional cloning techniques. Here, we report the implementation of a GATEWAY vector system enabling large-scale combination and investigation of candidate proteins in BiFC studies. We describe a set of 12 GATEWAY-compatible BiFC vectors that efficiently permit the combination of candidate protein pairs with every possible N-or C-terminal sub-fragment of S（CFP）3A or Venus, respectively, and enable the performance of multicolor BiFC （mcBiFC）. We used proteins of the plant molybdenum metabolism, in that more than 20 potentially interacting proteins are assumed to form the cellular molybdenum network, as a case study to establish the functionality of the new vectors. Using these vectors, we report the formation of the molybdopterin synthase complex by interaction of Arabidopsis proteins Cnx6 and Cnx7 detected by BiFC as well as the simultaneous formation of Cnx6/Cnx6 and Cnx6/Cnx7 complexes revealed by mcBiFC. Consequently, these GATEWAY-based BiFC vector systems should significantly facilitate the large-scale investigation of complex regulatory networks in plant cells.

关键词： BiFC GATEWAY molybdenum cofactor nitrate reductase.

Epidermal growth factor receptor pathway substrate 8 (Eps8) expression in maturing testis

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Asian Journal of Andrology 2004年第3期6卷 195-203页

作者： Alain Wunsch Kai Strothmann manuela Simoni jorg Gromoll Eberhard Nieschlag C.marc Luetjens Institute of Reproductive medicine of the University D-48129 MunsterGermanyInstitute of Reproductive Medicine of the UniversityD-48129 MunsterGermanyInstitute of Reproductive Medicine of the UniversityD-48129 MunsterGermanyInstitute of Reproductive Medicine of the UniversityD-48129 MunsterGermanyInstitute of Reproductive Medicine of the UniversityD-48129 MunsterGermanyInstitute of Reproductive Medicine of the UniversityD-48129 MunsterGermany

Aim: Although epidermal growth factor receptors are expressed in the testes, whether they signal through epidermal growth factor receptor pathway substrate 8 (Eps8) is unknown. Here we evaluated the expression pattern of Eps8 in the maturing testis. methods: The expression of Eps8 was analysed by Northern blotting, immunocy-tochemistry and Western blotting in primary Sertoli cell cultures and in testicular tissue of rodents. Results: Eps8 is specifically expressed in gonocytes, Leydig and Sertoli cells of the neonatal rats and in Leydig and Sertoli cells of the adult rats and mice. Although gonocytes express Eps8, no signal was found in prepubertal or mature spermatogonia and the expression level of Eps8 in Sertoli cells increases with age. No regulation of Eps8 expression in primary immature rat Sertoli cells by Follicle stimulating hormone (FSH) was detected by Western blotting. Conclusion: Eps8 seems to be involved in the growth factor-controlled regulation of cell proliferation and differentiation in the seminiferous epithelium. Eps8 is a possible marker for gonocytes and in Sertoli cells it could be involved in crosstalk with other growth factor pathways.

关键词： gene expression isoform Sertoli cell gonocyte

Epidemiology andinteractions of Human Immunodeficiency Virus - 1 and Schistosoma mansoni in sub-SaharanAfrica

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Infectious Diseases of Poverty 2013年第1期2卷 6-16页

作者： Humphrey D mazigo Fred Nuwaha Shona Wilson Safari m Kinunghi Domenica morona Rebecca Waihenya jorg Heukelbach David W Dunne Department of medical Parasitology and Entomology School of MedicineCathollc University of Health and Aled SclencesP.O.Box 1464MwanzaTanzanla Department of Environmental Health and Communicable Disease Control School of Public HealthCollege of Health SclencesMakerere UniversiyP.O.Box 7072KampalaUganda National Institute for medical.Research Mwanza Research CentreP.Q.Box 1462MwanzaTanzanla Department of Pathology University of CambidgeTennis Court RoadCambridge CB21OPUK Department of Zoology Faculty of SclenceJomo Kenyatta University of Agriculture and TechnologyP.O.Box ***NairobiKenya Department of Community Health School of MedicineFederal University of CearaCearaBail

Human Immunodeficiency Virus-1/AIDS and Schis tosoma mansoni are widespread in sub Saharan Africa and co-infection occurs *** the early 1990s,it has been suggested that the two infections may interact and potentiate the effects of each other within co-infected human ***,S mansoni infection has been suggested to be a risk factor for HIV transmission and progression in *** so,it would follow that mass deworming could have beneficial effects on HIV-1 transmission *** epidemiology of HIV in African countries is changing,shifting from urban to rural areas where the prevalence of Schistosoma mansoni is high and public health services are *** the other side,the consequent pathogenesis of HIV-1/*** co-infection remains *** we give an account of the epidemiology of HIV-1 and ***,discuss co-infection and possible biological causal relationships between the two infections,and the potential impact of praziquantel treatment on HIV-1 viral loads,CD4^(+) counts and CD4^(+)/CD8^(+) *** review of the available literature indicates that there is evidence to support the hypothesis that *** infections can influence the replication of the HIV-1,cel-to-cell transmission,as well as increase HIV progression as measured by reduced CD4^(+) T lymphocytes *** so,then deworming of HIV positive individuals living in endemic areas may impact on HV-1 viral loads and CD4^(+) T lymphocyte counts.

关键词： Schitosoma mansoni HIV-1 Co-infections Immunological interactions Deworming

One-step preparation of a novel graphitic biochar/Cu^(0)/Fe_(3)O_(4) composite using CO_(2)-ambiance pyrolysis to activate peroxydisulfate for dye degradation

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Journal of Environmental Sciences 2023年第3期125卷 26-36页

作者： Yangfan Yuan Changai Zhang Chenhao Zhao BingWang Xiaozhi Wang Bin Gao Shengsen Wang jorg Rinklebe College of Environmental Science and Engineering Yangzhou UniversityYangzhou 225127China Joint International Research Laboratory of Agriculture and Agri-Product Safety of ministry of Education of China Yangzhou UniversityYangzhou 225127China Jiangsu Collaborative Innovation Center for Solid Organic Waste Resource Utilization Yangzhou 225127China School of Environmental and Natural Resources Zhejiang University of Science and TechnologyHangzhou 310023China School of Resources and Environmental Engineering Guizhou UniversityGuiyang 550025China Department of Agricultural and Biological Engineering University of FloridaGainesville FL32611USA University of Wuppertal School of Architecture and Civil EngineeringInstitute of Foundation EngineeringWaterand Waste-ManagementLaboratory of Soil-and Groundwater-ManagementWuppertal 42285Germany Department of Environment Energy and GeoinformaticsSejong UniversityGuangjin-GuSeoul 05006South Korea

Herein,a one-step co-pyrolysis protocol was adopted for the first time to prepare a novel pyrogenic carbon-Cu^(0)/Fe_(3)O_(4)heteroatoms (FCBC) in CO_(2)ambiance to discern the roles of each component in PDS *** co-pyrolysis,CO_(2)catalyzed formation of reducing gases by biomass which facilitated reductive transformation of Fe^(3+)and Cu^(2+)to Cu^(0)and Fe_(3)O_(4),*** to the analysis,the resulting metal (oxide) catalyzed graphitization of biocharand decomposition of volatile substances resulting in an unprecedented surface area (1240 m^(2)/g).The resulting FCBC showed greater structural defects and less electrical *** experiments indicated that Rhodamine B (RhB) degradation by FCBC (100%) was superior to Fe_(3)O_(4)(50%) and Cu^(0)/Fe_(3)O_(4)(76.4%) in persulfate (PDS) system,which maintained reasonable efficiency (75.6%-63.6%) within three *** reactive oxygen species (ROS) associated with RhB degradation was identified by an electron paramagnetic resonance and confirmed by scavenging *** degradation invoked both(sulfate and dominantly hydroxyl) radical and non-radical (singlet oxygen,^(1)O_(2)) *** FCBC,Cu^(0)can continuously react with Fe^(3+)in Fe_(3)O_(4)to generate larger quantities of Fe^(2+),and both Cu^(0)and Fe^(2+)activated PDS to yield sulfate radicals which was quickly converted to hydroxyl ***,Cu^(0)/Cu^(2+)could complex with PDS to form a metastable complex,which particularly contributed to1O_(2)*** cascade reactions by FCBC were reinforced by carbonyl group of biochar and favorable electron transfer *** work highlighted a new approach to prepare a magnetic and environment-benign heterogonous catalyst to remove organic pollutants in water.

关键词： Biochar Singlet oxygen Persulfate Water pollution Waste biomass

A New β-Estradiol-lnducible Vector Set that Facilitates Easy Construction and Efficient Expression of Transgenes Reveals CBL3-Dependent Cytoplasm to Tonoplast Translocation of CIPK5

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molecular Plant 2013年第6期6卷 1814-1829页

作者： Kathrin Schliicking Kai H. Edel maria m. Drerupa Philipp Koster Christian Eckert Leonie Steinhorst Rainer Waadt Oliver Batistic jorg Kudla Institut for Biologie und Biotechnologie der Pflanzen Universit~t MQnster Schlossplatz 4 48149 MQnster Gel'many _ . _ Present address: University of California San Diego Division of Biological Sciences Cell and Developmental Biology Section 9500 Gilman Drive #0116 LaJolla CA 92093-0116 USA

Transient and stable expression of transgenes is central to many investigations in plant biology research. Chemical regulation of expression can circumvent problems of plant lethality caused by constitutive overexpression or allow inducible knock （out/down） approaches. Several chemically inducible or repressible systems have been described and successfully applied. However, cloning and application-specific modification of most available inducible expression systems have been limited and remained complicated due to restricted cloning options. Here we describe a new set of 57 vectors that enable transgene expression in transiently or stably transformed cells. All vectors harbor a synthetically optimized XVE expression cassette, allowing I^-estradiol mediated protein expression. Plasmids are equipped with the reporter genes GUS, GFP, mCherry, or with HA and Strepll epitope tags and harbor an optimized multiple cloning site for flexible and simple clon- ing strategies. moreover, the vector design allows simple substitution of the driving promoter to achieve tissue-specificity or to modulate expression ranges of inducible transgene expression. We report details of the kinetics and dose-dependence of expression induction in Arabidopsis leaf mesophyll protoplasts, transiently transformed Nicotiana benthamiana leaves, and stably transformed Arabidopsis plants. Using these vectors, we investigated the influence of CBL （Calcineurin B-like） protein expression on the subcellular localization of CIPKs （Calcineurin B-like interacting protein kinases）. These analyses uncovered that induced co-expression of CBL3 is fully sufficient for dynamic translocation of CIPK5 from the cytoplasm to the tonoplast. Thus, the vector system presented here facilitates a broad range of research applications.

关键词： vector inducible gene expression βestradiol Arabidopsis CBL CIPK.

Detection and characterization of murine colitis and carcinogenesis by molecularly targeted contrast-enhanced ultrasound

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World Journal of Gastroenterology 2017年第16期23卷 2899-2911页

作者： markus Brückner Jan Heidemann Tobias m Nowacki Friederike Cordes jorg Stypmann Philipp Lenz Faekah Gohar Andreas Lügering Dominik Bettenworth Department of medicine B University Hospital of MünsterD-48149 MünsterGermany Department of Gastroenterology Klinikum Bielefeld MitteAcademic Teaching HospitalUniversity of MünsterD-33604 BielefeldGermany Department of Cardiology and Angiology University Hospital of MünsterD-48149 MünsterGermany Department of Palliative Care University Hospital of MünsterD-48149 MünsterGermany Department of Paediatric Rheumatology and Immunology University Hospital of MünsterD-48149 MünsterGermany medical Care Center Portal 10 D-48155 MünsterGermany

AIm To study mucosal addressin cellular adhesion molecule-1(mAd CAm-1) and vascular endothelial growth factor(VEGF)-targeted contrast enhanced ultrasound(CEUS) for the assessment of murine colitis and carcinogenesis. mETHODS C57BL/6 mice were challenged with 3% dextran sodium-sulfate(DSS) for three, six or nine days to study the development of acute colitis. Ultrasound was performed with and without the addition of unspecific contrast agents. mAd CAm-1-targeted contrast agent was used to detect and quantify mAd CAm-1 expression. Inflammatory driven colorectal azoxymethane(AOm)/DSS-induced carcinogenesis was examined on day 42 and 84 using VEGF-targeted contrast agent. Highly specific tissue echogenicity was quantified using specialized software. Sonographic findings were correlated to tissue staining, western blot analysis and immunohistochemistry to quantify the degree of inflammation and stage of carcinogenesis. RESULTS Native ultrasound detected increased general bowel wall thickening that correlated with more progressed and more severe DSS-colitis(healthy mice: 0.3 mm ± 0.03 vs six days DSS: 0.5 mm ± 0.2 vs nine days DSS: 0.6 mm ± 0.2, P moreover, these sonographic findings correlated well with clinical parameters such as weight loss(r2 = 0.74) and histological damage(r2 = 0.86)(P murine colitis, CEUS targeted against mAd CAm-1 detected and differentiated stages of mild, moderate and severe colitis via calculation of mean pixel contrast intensity in decibel(9.6 d B ± 1.6 vs 12.9 d B ± 1.4 vs 18 d B ± 3.33, P mploying the AOm/DSSinduced carcinogenesis model, tumor development was monitored by CEUS targeted against VEGF and detected a significantly increased echogenicity in tumors as compared to adjacent healthy mucosa(healthy mucosa, 1.6 d B ± 1.4 vs 42 d, 18.2 d B ± 3.3 vs 84 d, 18.6 d B ± 4.9, P mmunohistochemistry findings(VEGF-posi

关键词： Colitis Dextran sodium-sulfate AOmDSS Carcinogenesis Ultrasound Contrast-enhanced ultrasound Vascular endothelial growth factor mucosal addressin cellular adhesion molecule-1

Effects of Exposure to GSm mobile Phone Base Station Signals on Salivary Cortisol,Alpha-Amylase,and Immunoglobulin A

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Biomedical and Environmental Sciences 2010年第3期23卷 199-207页

作者： CHRISTOPH AUGNER GERHARD W.HACKER GERD OBERFELD mATTHIAS FLORIAN WOLFGANG HITZL jorg HUTTER GERNOT PAUSER IGGmB-Research Institute for Frontier Questions of medicine and Biotechnology Salzburg Federal Hospital-University Clinics of the Paracelsus Medical Private UniversitySalzburg Federal Clinics(SALK)SalzburgAustria Public Health Department Salzburg SalzburgAustria Research office Paracelsus Medical Private UniversityComma SalzburgAustria University Clinic of Surgery Salzburg Federal Hospital University Clinics of the Paracelsus Medical Private UniversitySalzburg Federal Clinics(SALK)SalzburgAustria University Clinic for Anesthesiology Perioperative Medicine and General Intensive CareSalzburg Federal Hospital-University Clinics of the Paracelsus Medical Private UniversitySalzburg Federal Clinics(SALK)SalzburgAustria

Objective The present study aimed to test whether exposure to radiofrequency electromagnetic fields （RF-EmF） emitted by mobile phone base stations may have effects on salivary alpha-amylase, immunoglobulin A （IgA）, and cortisol levels. methods Fifty seven participants were randomly allocated to one of three different experimental scenarios （22 participants to scenario 1, 26 to scenario 2, and 9 to scenario 3）. Each participant went through five 50-minute exposure sessions. The main RF-EmF source was a GSm-900-mHz antenna located at the outer wall of the building. In scenarios 1 and 2, the first, third, and fifth sessions were ＂low＂（median power flux density 5.2 μW/m^2） exposure. The second session was ＂high＂（2126.8 μW/m^2）, and the fourth session was ＂medium＂（153.6 μW/m^2） in scenario 1, and vice versa in scenario 2. Scenario 3 had four ＂low＂ exposure conditions, followed by a ＂high＂ exposure condition. Biomedical parameters were collected by saliva samples three times a session. Exposure levels were created by shielding curtains. Results In scenario 3 from session 4 to session 5 （from ＂low＂ to ＂high＂ exposure）, an increase of cortisol was detected, while in scenarios 1 and 2, a higher concentration of alpha-amylase related to the baseline was identified as compared to that in scenario 3. IgA concentration was not significantly related to the exposure. Conclusions RF-EmF in considerably lower field densities than ICNIRP-gnidelines may influence certain psychobiological stress markers.

关键词： GSm base stations mobile phone Salivary IgA Alpha amylase Cortisol Radiofrequency electromagnetic fields (RF-EmF)

Improving the haploidization toolbox:maternal factors take the stage

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molecular Plant 2023年第4期16卷 651-653页

作者： maria Flores-Tornero Helena Sapeta jorg D.Becker Instituto de Tecnologia Quimica e Biologica Universidade Nova de LisboaAv.daRepublica2780-157OeirasPortugal

Plant breeders have a great interest in obtaining plants that are genetically homogeneous as quickly as possible,with high efficiency and at low *** approaches,including selfing or backcrossing,cannot provide *** technologies,however,offer a solution and are therefore the subject of intensive research,in particular during the last decade.

关键词： breed homogeneous quickly

Effect of toasting grain silages from field peas(Pisum sativum)and field beans(Vicia faba)on in vitro gas production,methane production,and post-ruminal crude protein content

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Animal Nutrition 2020年第3期6卷 342-352页

作者： martin Bachmann Christian Kuhnitzsch Sebastian michel Annabel Thierbach mandy Bochnia jorg m.Greef Siriwan D.martens Olaf Steinhofel Annette Zeyner Institute of Agricultural and Nutritional Sciences Martin Luther University Halle-WittenbergHalle(Saale)06120Germany Saxon State Office for Environment Agriculture and GeologyKollitsch04886Germany Julius Kühn Institute Federal Research Centre for Cultivated PlantsInstitute for Crop and Soil ScienceBraunschweig38116Germany

Legume grains such as field peas and field beans can be produced on a local level,and may be reliable sources of dietary protein and energy apart from common soybean and rapeseed *** ruminants,protein,starch,and carbohydrates from peas and field beans are fermented in large part before reaching the small *** objective of this study was to evaluate the effects of a combination of ensiling and hydro-thermic treatment(i.e.,toasting at 160℃for 30 min)of grains of peas and field beans on the concentrations of post-ruminal crude protein(PRCP)and rumen-undegraded protein(RUP).moreover,24-h gas production and methane production were *** this,an in vitro batch culture system with ruminal fluid from sheep was ***-undegraded protein was determined using the Streptomyces griseus protease *** electron micrographs were used to visualize morphological changes of starch granules and their joint matrices in peas and field beans after ensiling,toasting,or a combination of *** pea grains contained crude protein(CP)at 199 g/kg Dm,PRCP at 155 g/kg Dm at a ruminal passage rate of 0.08/h(Kp8),RUP at 33 g/kg Dm at Kp8,and starch at 530 g/kg *** field beans contained CP at 296 g/kg Dm,PRCP at 212 g/kg Dm at Kp8,RUP at 54 g of/kg Dm at Kp8,and starch at 450 g/kg *** PRCP did not considerably differ among native and treated peas or field *** in the peas,RUP at Kp8 increased after ensiling by 10 g/kg Dm(i.e.,30%;Pmethane production were not affected by any treatment,and scanning electron micrographs did not reveal structural changes on the starches doubtless of any *** seemed to be more affected by treatment with ensiled+toasted peas than with ensiled+toasted field beans,but starches and other carbohydrates from both legumes remained unaffec

关键词： Legume grain silage Toasting In vitro gas production Protein evaluation Streptomyces griseus protease test