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Silk fibroin as an additive for cell-free protein synthesis

Synthetic and Systems biotechnology 2020年第3期5卷 145-154页

作者： Marilyn S.Lee Chia-Suei Hung daniel A.Phillips Chelsea C.Buck Maneesh K.Gupta Matthew W.Lux US Army Combat Capabilities Development Command Chemical and biological Center 8567 Ricketts Point RoadAberdeen Proving GroundMD21010USA US Air Force Research Laboratory 217912th St.B652/R122 Wright-Patterson Air Force BaseOH45433USA US Naval Research Laboratory Center for bio/Molecular Science and Engineering Bldg.42Room 3034555 Overlook Ave.WashingtonDC 20375UES Inc.4401 Dayton Xenia Rd.BeavercreekOH 45432USA

Cell-free systems contain many proteins and metabolites required for complex functions such as transcription and translation or multi-step metabolic *** into expanding the delivery of these systems by drying or by embedding into other materials is enabling new applications in sensing,point-of-need manufacturing,and responsive ***,silk fibroin from the silk worm,Bombyx mori,has received attention as a protective additive for dried enzyme formulations and as a material to build biocompatible hydrogels for controlled localization or delivery of biomolecular *** this work,we explore the effects of silk fibroin as an additive in cell-free protein synthesis(CFPS)*** of silk fibroin on CFPS activity and stability after drying,as well as the potential for incorporation of CFPS into hydrogels of crosslinked silk fibroin are *** find that simple addition of silk fibroin increased productivity of the CFPS reactions by up to 42%,which we attribute to macromolecular crowding ***,we did not find evidence that silk fibroin provides a protective effects after drying as previously described for purified ***,the enzymatic crosslinking transformations of silk fibroin typically used to form hydrogels are inhibited in the presence of the CFPS reaction *** attempts did not impact CFPS activity,but did yield localized protein aggregates rather than a *** discuss the mechanisms at play in these results and how the silk fibroin-CFPS system might be improved for the design of cell-free devices.

关键词： Cell-free systems Cell-free protein synthesis CFPS Silk fibroin Preservation

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Reaction mechanism of aqueous-phase conversion of γ-valerolactone(GVL) over a Ru/C catalyst

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Journal of Energy Chemistry 2016年第6期25卷 1008-1014页

作者： Abigail Rozenblit Adam J.Avoian Qiaohua Tan Tawan Sooknoi daniel E.Resasco School of Chemical Biological and Materials Engineering (CBME) and Center for Interfacial Reaction Engineering (CIRE) University of Oklahoma

The present work explores the reaction pathways of γ-valerolactone(GVL) over a supported ruthenium catalyst. The conversion of GVL in aqueous phase over a 5% Ru/C catalyst was investigated in a batch reactor operating at 463 K under 500–1000 psi of H2. The main reaction products obtained under these conditions were 2-butanol(2-BuOH), 1,4-pentanediol(1,4-PDO), 2-methyltetrahydrofuran(2-MTHF) and 2-pentanol(2-PeOH). A complete reaction network was developed, identifying the primary and/or secondary products. In this reaction network, production of 2-BuOH via decarbonylation of a ring-opened surface intermediate CH3CH(O*)–(CH2)2–CO*is clearly the dominant pathway. From the evolution of products as a function of reaction time and theoretical(DFT) calculations, a mechanism for the formation of intermediates and products is proposed. The high sensitivity of 2-BuOH production to the presence of CO, compared to a much lower effect on the production of the other products indicates that the sites responsible for decarbonylation are particularly prone to CO adsorption and poisoning. Also, since the decarbonylation rate is not affected by the H2 pressure it is concluded that the direct decarbonylation path of the CH3CH(O*)–(CH2)2–CO*intermediate does not required a previous dehydrogenation step, as is the case in decarbonylation of short alcohols.

关键词： γ-Valerolactone GVL Butanol Pentanol Pentanediol Ring-opening Hydrogenolysis Hydrogenation Decarbonylation Ru/C catalyst bio-oil upgrading in aqueous phase

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Predicting Stem Windthrow Probability in a Northern Hardwood Forest Using a Wind Intensity bio-Indicator Approach

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Open Journal of Forestry 2012年第2期2卷 77-87页

作者： Philippe Nolet Frédérik Doyon daniel Bouffard Insitut des Sciences de la Forêt tempérée Ripon Canada

Unlike fire or insect outbreaks, for which a suppression program can be implemented, it is impossible to prevent a windstorm event or stop it while it is occurring. Reducing stand susceptibility to windstorms requires a good understanding of the factors affecting this susceptibility. Distinct species- and size-related differences in stem windthrow susceptibility are difficult to obtain because it is impossible to distinguish their relative effects from those of wind intensity. Using a damage assessment database (60 20-metre radius plots) acquired after an exceptional wind storm in Western Quebec in 2007, we developed an approach in which proportions of windthrown sugar maple poles were used as bio-indicators of wind intensities affecting the plots. We distinguished between single and interactive effects of wind intensity, species, stem size, and local basal area on stem windthrow susceptibility. The best logistic regression model predicting stem windthrow included the wind intensity bio-indicator, species, basal area, and the species by diameter at breast height (DBH, 1.3 m) interaction. Stem windthrow probability generally increased with DBH and decreased with basal area. Species wind-firmness was ordered as: yellow birch > sugar maple = eastern hemlock = American beech > ironwood > basswood = other hardwoods = other softwoods. Our method remained an indirect method of measuring wind intensity and its real test would require a comparison with anemometer measurements during a windstorm. Despite its indirect nature, the method is both simple and ecologically sound. Hence, it opens the door to conducting similar windthrow studies in other ecosystems.

关键词： Windthrow Prediction Species Wind-Firmness Tolerant Hardwood Forests Silviculture Guidelines

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Molecularly specific detection of bacterial lipoteichoic acid for diagnosis of prosthetic joint infection of the bone

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Bone Research 2018年第2期6卷 189-196页

作者： Julie E.Pickett John M.Thompson Agnieszka Sadowska Christine Tkaczyk Bret R.Sellman Andrea Minola Davide Corti Antonio Lanzavecchia Lloyd S.Miller daniel LJ Thorek Division of Nuclear Medicine and Molecular Imaging Department of Radiology and Radiological ScienceJohns Hopkins University School of MedicineGaithersburgMDUSA Department of Orthopaedic Surgery Johns Hopkins University School of MedicineBaltimoreMD 21205USA Department of Infectious Disease MedImmune LLCGaithersburgMDUSA Humabs bio Med SA BellinzonaSwitzerland Institute for Research in biomedicine Universitàdella Svizzera italianaBellinzonaSwitzerland Department of Dermatology Johns Hopkins University School of MedicineBaltimoreMD 21205USA Division of Infectious Disease Department of MedicineJohns Hopkins University School of MedicineBaltimoreMDUSA Cancer Molecular and Functional Imaging Program Department of OncologySidney Kimmel Comprehensive Cancer CenterJohns Hopkins University School of MedicineBaltimoreMD 21205USA

Discriminating sterile inflammation from infection, especially in cases of aseptic loosening versus an actual prosthetic joint infection, is challenging and has significant treatment implications. Our goal was to evaluate a novel human monoclonal antibody（mAb） probe directed against the Gram-positive bacterial surface molecule lipoteichoic acid（LTA）. Specificity and affinity were assessed in vitro. We then radiolabeled the anti-LTA mAb and evaluated its effectiveness as a diagnostic imaging tool for detecting infection via immuno PET imaging in an in vivo mouse model of prosthetic joint infection（PJI）. In vitro and ex vivo binding of the anti-LTA mAb to pathogenic bacteria was measured with Octet, ELISA, and flow cytometry. The in vivo PJI mouse model was assessed using traditional imaging modalities, including positron emission tomography（PET） with [^（18）F]FDG and [^（18）F]Na F as well as X-ray computed tomography（CT）, before being evaluated with the zirconium-89-labeled antibody specific for LTA（[^（89）Zr]SAC55）.The anti-LTA mAb exhibited specific binding in vitro to LTA-expressing bacteria. Results from imaging showed that our model could reliably simulate infection at the surgical site by bioluminescent imaging, conventional PET tracer imaging, and bone morphological changes by CT. One day following injection of both the radiolabeled anti-LTA and isotype control antibodies, the anti-LTA antibody demonstrated significantly greater（P 〈 0.05） uptake at S. aureus-infected prosthesis sites over either the same antibody at sterile prosthesis sites or of control non-specific antibody at infected prosthesis sites. Taken together, the radiolabeled anti-LTA mAb, [^（89）Zr]SAC55, may serve as a valuable diagnostic molecular imaging probe to help distinguish between sterile inflammation and infection in the setting of PJI. Future studies are needed to determine whether these findings will translate to human PJI.

关键词： Zr Molecularly specific detection bacterial lipoteichoic acid diagnosis prosthetic infection bone LTA SAC PET FDG tracer

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Transoral outlet reduction:Outcomes of endoscopic Roux-en-Y gastric bypass revision in 284 patients at a community practice

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World Journal of Gastrointestinal Endoscopy 2023年第10期15卷 602-613页

作者： daniel B Maselli Vibhu Chittajallu Chase Wooley Areebah Waseem daniel Lee Michelle Secic Lauren LDonnangelo Brian Coan Christopher E McGowan Clinical Research True You Weight LossAtlantaGA 30342United States Department of Gastroenterology and Hepatology University Hospitals Cleveland Medical CenterClevelandOH 44106United States Clinical Research True You Weight LossCaryNC 27513United States Secic Statistical Consulting ClevelandOH 44106United States

BACKGROUND Transoral outlet reduction(tore)is a minimally invasive endoscopic revision of Roux-en-Y gastric bypass(RYGB)for weight recurrence;however,little has been published on its clinical implementation in the community *** To characterize the safety and efficacy of tore in the community setting for adults with weight recurrence after *** This is a retrospective cohort study of argon plasma coagulation and purse-string suturing for gastric outlet reduction in consecutive adults with weight recurrence after RYGB at a single community center from September 2020 to September *** were provided longitudinal nutritional support via virtual *** primary outcome was total body weight loss(TBWL)at twelve months from *** outcomes included TBWL at three months and six months;excess weight loss(EWL)at three,six,and twelve months;twelve-month TBWL by obesity class;predictors of twelve-month TBWL;rates of post-tore stenosis;and serious adverse events(SAE).Outcomes were reported with descriptive *** Two hundred eighty-four adults(91.9%female,age 51.3 years,body mass index 39.3 kg/m^(2))underwent tore an average of 13.3 years after *** pre-and post-tore outlet diameter was 35 mm and 8 mm,*** was 11.7%±4.6%at three months,14.3%±6.3%at six months,and 17.3%±7.9%at twelve *** was 38.4%±28.2%at three months,46.5%±35.4%at six months,and 53.5%±39.2%at twelve *** number of follow-up visits attended was the strongest predictor of TBWL at twelve months(R^(2)=0.0139,P=0.0005).Outlet stenosis occurred in 11 patients(3.9%)and was successfully managed with endoscopic *** was one instance of post-procedural nausea requiring overnight observation(SAE rate 0.4%).CONCLUSION When performed by an experienced endoscopist and combined with longitudinal nutritional support,purse-string tore is safe and effective in the community setting for adults with weight recurrence after RYGB.

关键词： Transoral outlet reduction Purse-string Roux-en-Y gastric bypass Obesity Endoscopic revision Weight recurrence Gastrojejunal anastomosis

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蛇形画廊

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城市环境设计 2019年第2期118卷 44-53页

作者： Aaron Powers Alice Cladet Claire Thomas daniel Sundlin Jakob Lange Kai-Uwe Bergmann Kristian Hindsberg Kristoffer Negendahl Lorenz Krisai Maria Holst Maxwell Moriyama Rune Hansen Tianze Li tore Banke Wells Barber Iwan Baan Derek Shapton 陈子钰

对于2016年的蛇形画廊展厅,我们尝试设计一种包含多个通常意义上的对立面的结构:一种既形态自由又结构严谨、既模块化又具有雕塑感、既透明又不透明、既似盒子又难以名状的结构。我们决定使用建筑最基本的元素之一:砖墙。并非粘土砖或... 详细信息

对于2016年的蛇形画廊展厅,我们尝试设计一种包含多个通常意义上的对立面的结构:一种既形态自由又结构严谨、既模块化又具有雕塑感、既透明又不透明、既似盒子又难以名状的结构。我们决定使用建筑最基本的元素之一:砖墙。并非粘土砖或石块砖——这座墙是由

关键词：内部空间玻璃纤维

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Handheld endomicroscope using a fiber-optic harmonograph enables real-time and in vivo confocal imaging of living cell morphology and capillary perfusion

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Microsystems & Nanoengineering 2020年第1期6卷 449-459页

作者： Kyungmin Hwang Yeong-Hyeon Seo daniel YKim Jinhyo Ahn Soyoung Lee Kyung Hee Han Koun-Hee Lee Sangyong Jon Pilhan Kim Kate EYu Hyungsin Kim Shin-Hyuk Kang Ki-Hun Jeong Department of bio and Brain Engineering KAIST and KAIST Institute of Health Science and TechnologyDaejeon 34141Republic of Korea Graduate School of Nanoscience and Technology KAIST and KAIST Institute of Health Science and TechnologyDaejeon 34141Republic of Korea Department of biological Sciences KAIST and KAIST Institute for the BioCenturyDaejeon 34141Republic of Korea Hyunjoo In-Tech Seoul 08390Republic of Korea VPIX Medical IncDeajeon 34141Republic of Korea Graduate School of Medical Science and Engineering Daejeon 34141Republic of Korea Department of Neurosurgery Korea University Anam HospitalKorea University MedicineSeoul 02842Korea

Confocal laser endomicroscopy provides high potential for noninvasive and in vivo optical biopsy at the cellular ***,we report a fully packaged handheld confocal endomicroscopic system for real-time,high-resolution,and in vivo cellular imaging using a Lissajous scanning fiber-optic *** endomicroscopic system features an endomicroscopic probe with a fiber-optic harmonograph,a confocal microscope unit,and an image signal *** fiber-optic harmonograph contains a single mode fiber coupled with a quadrupole piezoelectric tube,which resonantly scans both axes at~1 kHz to obtain a Lissajous *** fiber-optic harmonograph was fully packaged into an endomicroscopic probe with an objective *** endomicroscopic probe was hygienically packaged for waterproofing and disinfection of medical instruments within a 2.6-mm outer diameter stainless tube capable of being inserted through the working channel of a clinical *** probe was further combined with the confocal microscope unit for indocyanine green imaging and the image signal processor for high frame rate and high density Lissajous *** signal processing unit delivers driving signals for probe actuation and reconstructs confocal images using the auto phase matching process of Lissajous fiber *** confocal endomicroscopic system was used to successfully obtain human in vitro fluorescent images and real-time ex vivo and in vivo fluorescent images of the living cell morphology and capillary perfusion inside a single mouse.

关键词： fiber processor confocal

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Development of a chromatographic lateral flow immunoassay for detection of African swine fever virus antigen in blood

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Animal Diseases 2022年第3期2卷 139-153页

作者： daniel W.Madden Sun-Young Sunwoo Natasha N.Gaudreault Jessie D.Trujillo Igor Morozov Carmina Gallardo Juergen A.Richt Department of Diagnostic Medicine/Pathobiology College of Veterinary MedicineKansas State UniversityManhattanKSUSA Centro de Investigación en Sanidad Animal(CISA-INIA/CSIC) MadridSpain

African swine fever(ASF)is a highly lethal disease of domestic and wild swine caused by African swine fever virus(ASFV).The disease currently circulates in Africa,Europe,Asia and on the island of *** ongoing epizootics in Europe and Asia have produced millions of animal deaths and severe economic *** effective vaccine is available for ASF,making rapid and accurate detection of ASFV essential for disease mitigation *** available diagnostics for ASFV possess significant limitations related to assay performance,deployability,and/or turn-around time;therefore there is an unmet need for pen-side diagnostic tests with sufficient sensitivity and specificity.A chromatographic lateral flow immunoassay(LFIA)was developed for the detection of ASFV antigen in EDTA-treated whole blood using monoclonal antibodies targeting the viral p30 *** assay requires only water to perform and provides results in 25 min,making it well-suited for field *** LFIA was capable of detecting genotype I and genotype II strains of ASFV in EDTA blood from experimentally infected pigs at varying time-points after infection,though it was unable to detect a genotype X ASFV *** sensitivity correlated with clinical disease severity,body temperature,and viral DNA levels,and was over 90%in animals showing moderate to severe ASF-related symptoms after challenge with virulent genotype II *** LFIA also showed a robust diagnostic specificity of over 98%,which is essential to field testing for a high consequence to foregin animal *** LFIA targeting the viral p30 protein can reliably detect ASFV in whole blood from animals showing moderate to severe clinical signs of infection with virulent genotype I and II isolates,making it a promising candidate for use as a field-deployable antigen detection *** evaluation using field samples and different virus strains is required to further assess the utility of this rapid diagnostic test.

关键词： African swine fever Lateral flow immunoassay Point of care diagnostics

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The Effect of Indigenous Growth Media on Allanblackia parviflora A. Chev in Ghana

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Open Journal of Soil Science 2016年第5期6卷 89-97页

作者： Edward Yeboah daniel Aninagyei Ofori Theresa Peprah Ramni Harmanjeet Jamnadass Alain Tsobeng CSIR-Soil Research Institute Academy Post Office Kumasi Ghana CSIR-Forestry Research Institute of Ghana University Post Office KNUST Kumasi Ghana World Agroforestry Centre Nairobi Kenya World Agroforestry Centre Yaoundé Cameroon

Allanblackia parviflora A. Chev. also called vegetable tallow tree provides a variety of non-timber forest products of great importance to rural households including shade, timber, medicine and seed oil but attempts have not been made to improve the tree species and increase its production. Consequently, the species is being threatened due to unsustainable exploitation and poor regeneration and cultivation appears as the only viable option. In order to cultivate the species at meaningful scale, it is necessary to establish the optimum range of environmental factors that influence its propagation and growth. This study was therefore designed to investigate Allanblackia growth parameters and bio-accumulation under different growth media in a greenhouse study. The media were: 1) TS = top soil alone, 2) AB soil = Allanblackia soil alone, 3) TS + H = Top soil alone + humus, 4) AB + TS = Allanblackia soil alone + Top soil alone and 5) SAB = Sterilized Allanblackia soil alone. Each treatment was replicated three times in a complete randomized design. The experiment lasted for 18 months. Results showed that Fe was the micronutrient that accumulated greatest in the plant tissue. Among the treatments, Allanblackia soil showed the highest accumulation of Zn in the plant tissue with the top soil showing the least (7.67 mg·kg-l). Humus contributed largely to the bio-accumulation of Cu in the plant tissue. bio-accumulation of manganese in the plant tissue ranged from 13.30 mg·kg-l to 207 mg·kg-l suggesting difference in manganese absorption by Allanblackia as influenced by the treatments. The growth parameters of Allanblackia parviflory were impacted differently by the growth media. The result was however controversial since no differences were found between growth of seedlings in sterilized Allanblackia soil and Allanblackia soil.

关键词： Allanblackia bio-Accumulation Growth Media Mycorrhiza

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Chemoenzymatic Synthesis and Molecular biology: Searching an Access to Single form Glycoproteins

Chemoenzymatic Synthesis and Molecular Biology: Searching an...

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8th Chinese International Peptide Symposium

作者： Carlo UNVERZAGT Stefano MEZZATO Manuela SCHAFFRATH daniel VARON bioorganische Chemie Gebude NWI Universitt Bayreuth95440 BayreuthGermany

Recent efforts to obtain functional glycoproteins as a single glycoform have been a challenge for interdisciplinary chemical research. Approaches to reduce the diversity of natural glycoproteins by glycan remodeling, transglycosy-lation, reverse proteolysis or native chemical ligation have been proposed. The most promising approach for the synthesis of complex type N-glycoproteins appears the native chemical ligation of glycoprotein fragments. As a model glycoprotein we chose RNase because the reference enzyme can be readily obtained in a glycosylated and a non-gly-cosylated form from natural sources and structure-activity studies indicated that the enzymatic activity is dependent on the glycosylation pattern. Glycosylated RNase B was retrosynthetically disconnected into two fragments ranging

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