bluuebunny/crossref_metadata_2025_split huggingface dataset:
Der Einbau der neuen Hilfsbrücke ZH 31 an der Eisenbahnüberführung Stauffenbergallee in Dresden
Authors: Tusche Jens 
Published: October 2017

AbstractIm Zuge der Eisenbahnstrecke Görlitz–Dresden befindet sich die Eisenbahnüberführung über die Stauffenbergallee in Dresden. Diese Brücke wurde bedingt durch ihren schlechten Zustand durch Hilfsbrücken ersetzt. Dabei handelt es sich um einen der ersten Einsätze der neu entwickelten Zwillingsträgerhilfsbrücke ZH 31, die Stützweiten von 28,80–31,20 m ermöglicht und damit das Zwillingsträgerhilfsbrückensortiment der Deutschen Bahn von Stützweiten von bisher bis 26,40 auf 31,20 m ergänzt.
The Role of Independence and Stationarity in Probabilistic Models of Binary Choice
Authors: Regenwetter Michel , Davis‐Stober Clintin P. 
Published: October 2017

AbstractAfter more then 50 years of probabilistic choice modeling in economics, marketing, political science, psychology, and related disciplines, theoretical and computational advances give scholars access to a sophisticated array of modeling and inference resources. We review some important, but perhaps often overlooked, properties of major classes of probabilistic choice models. For within‐respondent applications, we discuss which models require repeated choices by an individual to be independent and response probabilities to be stationary. We show how some model classes, but not others, are invariant over variable preferences, variable utilities, or variable choice probabilities. These models, but not others, accommodate pooling of responses or averaging of choice proportions within participant when underlying parameters vary across observations. These, but not others, permit pooling/averaging across respondents in the presence of individual differences. We also review the role of independence and stationarity in statistical inference, including for probabilistic choice models that, themselves, do not require those properties. Copyright © 2017 John Wiley & Sons, Ltd.
Comparison of platform host cell protein ELISA to process‐specific host cell protein ELISA
Authors: Gunawan Feny , Nishihara Julie , Liu Peter , Sandoval Wendy , Vanderlaan Marty , Zhang Heidi , Krawitz Denise 
Published: October 2017

AbstractDuring expression of biotherapeutic proteins, complex mixtures of additional proteins are also produced by normal expression machinery of the host cell (termed “host cell proteins,” or HCP). HCPs pose a potential impact to patient safety and product efficacy, and therefore must be well‐characterized and the ability of the process to clear these proteins must be demonstrated. Due to the complexity of HCP, the method(s) used for monitoring must be demonstrated to provide sufficient information about relevant proteins. The most commonly used analytical method for monitoring HCP is an enzyme‐linked immunosorbent assay (ELISA). To ensure development of a suitable HCP ELISA, careful selection of critical reagents (anti‐HCP antibodies and analytical standard) is crucial. During a recent major update to the manufacturing process of a biotherapeutic, we re‐evaluated the suitability of the existing HCP ELISA for monitoring the HCP population in the updated process. In the evaluation, we compared a process‐specific ELISA to a platform ELISA. Despite qualitative differences in the HCP profiles in 2D PAGE, LC‐MS/MS showed that the HCP populations in the two analytical standards were similar. The process‐specific HCP antibody had adequate HCP coverage, but was more sensitive to a few dominant proteins that were present in the upstream purification process. The platform HCP antibody had very broad coverage and additionally, could detect the majority of potential HCP impurities from this process. Furthermore, the platform HCP antibody was not biased toward a few dominant proteins and was more sensitive in the downstream purification process. Due to its broad HCP coverage and sensitivity, we conclude that our platform HCP ELISA method is superior to the process‐specific HCP ELISA method.
One‐step affinity capture and precipitation for improved purification of an industrial monoclonal antibody using Z‐ELP functionalized nanocages
Authors: Swartz Andrew R. , Xu Xuankuo , Traylor Steven J. , Li Zheng J. , Chen Wilfred 
Published: October 2017

AbstractProtein A chromatography has been identified as a potential bottleneck in the monoclonal antibody production platform, leading to increased interest in non‐chromatographic capture technologies. Affinity precipitation using environmentally responsive, Z‐domain‐elastin‐like polypeptide (Z‐ELP) fusion proteins has been shown to be a promising alternative. However, elevated temperature and salt concentrations necessary for precipitation resulted in decreased antibody monomer content and reduced purification capacity. To improve upon the existing technology, we reported an enhanced affinity precipitation of antibodies by conjugating Z‐ELP to a 25 nm diameter, self‐assembled E2 protein nanocage (Z‐ELP‐E2). The enlarged scale of aggregate formation and IgG‐triggered crosslinking through multi‐valent binding significantly outperformed traditional Z‐ELP‐based methods. In the current work, we sought to develop an affinity precipitation process capable of purifying industrial monoclonal antibodies (mAbs) at ambient temperature with minimal added salt. We discovered that the mAb‐nanocage complex aggregated within 10 min at room temperature without the addition of salt due to the enhanced multi‐valent cross‐linking. After precipitating out of solution, the complex remained insoluble under all wash buffers tested, and only resolubilized after a low pH elution. Through optimization of key process steps, the affinity precipitation yield and impurity clearance met or exceeded protein A chromatography performance with 95% yield, 3.7 logs host cell protein reduction, and >5 logs of DNA reduction from mAb cell culture. Because of the operational flexibility afforded by this one‐step affinity capture and precipitation process, the Z‐ELP‐E2 based approach has the potential to be a viable alternative to platform mAb purification.
Biocatalytic conversion of cycloalkanes to lactones using an in‐vivo cascade in Pseudomonas taiwanensis VLB120
Authors: Karande Rohan , Salamanca Diego , Schmid Andreas , Buehler Katja 
Published: October 2017

AbstractChemical synthesis of lactones from cycloalkanes is a multi‐step process challenged by limitations in reaction efficiency (conversion and yield), atom economy (by‐products) and environmental performance. A heterologous pathway comprising novel enzymes with compatible kinetics was designed in Pseudomonas taiwanensis VLB120 enabling in‐vivo cascade for synthesizing lactones from cycloalkanes. The respective pathway included cytochrome P450 monooxygenase (CHX), cyclohexanol dehydrogenase (CDH), and cyclohexanone monooxygenase (CHXON) from Acidovorax sp. CHX100. Resting (non‐growing) cells of the recombinant host P. taiwanensis VLB120 converted cyclohexane, cyclohexanol, and cyclohexanone to ϵ‐caprolactone at 22, 80–100, and 170 U gCDW−1, respectively. Cyclohexane (5 mM) was completely converted with a selectivity of 65% for ϵ‐caprolactone formation in 2 hr without accumulation of intermediate products. Promiscuity of the whole‐cell biocatalyst gave access to analogous lactones from cyclooctane and cyclodecane. A total product concentration of 2.3 g L−1 and a total turnover number of 36,720 was achieved over 5 hr with a biocatalyst concentration of 6.8 gCDW L−1.
Enzymatic synthesis of chiral amino‐alcohols by coupling transketolase and transaminase‐catalyzed reactions in a cascading continuous‐flow microreactor system
Authors: Gruber Pia , Carvalho Filipe , Marques Marco P. C. , O'Sullivan Brian , Subrizi Fabiana , Dobrijevic Dragana , Ward John , Hailes Helen C. , Fernandes Pedro , Wohlgemuth Roland , Baganz Frank , Szita Nicolas 
Published: October 2017

AbstractRapid biocatalytic process development and intensification continues to be challenging with currently available methods. Chiral amino‐alcohols are of particular interest as they represent key industrial synthons for the production of complex molecules and optically pure pharmaceuticals. (2S,3R)‐2‐amino‐1,3,4‐butanetriol (ABT), a building block for the synthesis of protease inhibitors and detoxifying agents, can be synthesized from simple, non‐chiral starting materials, by coupling a transketolase‐ and a transaminase‐catalyzed reaction. However, until today, full conversion has not been shown and, typically, long reaction times are reported, making process modifications and improvement challenging. In this contribution, we present a novel microreactor‐based approach based on free enzymes, and we report for the first time full conversion of ABT in a coupled enzyme cascade for both batch and continuous‐flow systems. Using the compartmentalization of the reactions afforded by the microreactor cascade, we overcame inhibitory effects, increased the activity per unit volume, and optimized individual reaction conditions. The transketolase‐catalyzed reaction was completed in under 10 min with a volumetric activity of 3.25 U ml−1. Following optimization of the transaminase‐catalyzed reaction, a volumetric activity of 10.8 U ml−1 was attained which led to full conversion of the coupled reaction in 2 hr. The presented approach illustrates how continuous‐flow microreactors can be applied for the design and optimization of biocatalytic processes.
Impact of carbon monoxide partial pressures on methanogenesis and medium chain fatty acids production during ethanol fermentation
Authors: Esquivel‐Elizondo Sofia , Miceli Joseph , Torres Cesar I. , Krajmalnik‐Brown Rosa 
Published: October 2017

AbstractMedium‐chain fatty acids (MCFA) are important biofuel precursors. Carbon monoxide (CO) is a sustainable electron and carbon donor for fatty acid elongation, since it is metabolized to MCFA precursors, it is toxic to most methanogens, and it is a waste product generated in the gasification of waste biomass. The main objective of this work was to determine if the inhibition of methanogenesis through the continuous addition of CO would lead to increased acetate or MCFA production during fermentation of ethanol. The effects of CO partial pressures (PCO; 0.08–0.3 atm) on methanogenesis, fatty acids production, and the associated microbial communities were studied in batch cultures fed with CO and ethanol. Methanogenesis was partially inhibited at PCO ≥ 0.11 atm. This inhibition led to increased acetate production during the first phase of fermentation (0–19 days). However, a second addition of ethanol (day 19) triggered MCFA production only at PCO ≥ 0.11 atm, which probably occurred through the elongation of acetate with CO‐derived ethanol and H2:CO2. Accordingly, during the second phase of fermentation (days 20–36), the distribution of electrons to acetate decreased at higher PCO, while electrons channeled to MCFA increased. Most probably, Acetobacterium, Clostridium, Pleomorphomonas, Oscillospira, and Blautia metabolized CO to H2:CO2, ethanol and/or fatty acids, while Peptostreptococcaceae, Lachnospiraceae, and other Clostridiales utilized these metabolites, along with the provided ethanol, for MCFA production. These results are important for biotechnological systems where fatty acids production are preferred over methanogenesis, such as in chain elongation systems and microbial fuel cells.
Metabolic engineering of Bacillus subtilis for production of D‐lactic acid
Authors: Awasthi Deepika , Wang Liang , Rhee Mun S. , Wang Qingzhao , Chauliac Diane , Ingram Lonnie O. , Shanmugam Keelnatham T. 
Published: October 2017

ABSTRACTPoly lactic acid (PLA) based plastics is renewable, bio‐based, and biodegradable. Although present day PLA is composed of mainly L‐LA, an L‐ and D‐ LA copolymer is expected to improve the quality of PLA and expand its use. To increase the number of thermotolerant microbial biocatalysts that produce D‐LA, a derivative of Bacillus subtilis strain 168 that grows at 50°C was metabolically engineered. Since B. subtilis lacks a gene encoding D‐lactate dehydrogenase (ldhA), five heterologous ldhA genes (B. coagulans ldhA and gldA101, and ldhA from three Lactobacillus delbrueckii) were evaluated. Corresponding D‐LDHs were purified and biochemically characterized. Among these, D‐LDH from L. delbrueckii subspecies bulgaricus supported the highest D‐LA titer (about 1M) and productivity (2 g h−1 g cells−1) at 37°C (B. subtilis strain DA12). The D‐LA titer at 48°C was about 0.6 M at a yield of 0.99 (g D‐LA g−1glucose consumed). Strain DA12 also fermented glucose at 48°C in mineral salts medium to lactate at a yield of 0.89 g g−1 glucose and the D‐lactate titer was 180 ± 4.5 mM. These results demonstrate the potential of B. subtilis as a platform organism for metabolic engineering for production of chemicals at 48°C that could minimize process cost.
A synthetic biology approach to transform Yarrowia lipolytica into a competitive biotechnological producer of β‐carotene
Authors: Larroude Macarena , Celinska Ewelina , Back Alexandre , Thomas Stephan , Nicaud Jean‐Marc , Ledesma‐Amaro Rodrigo 
Published: October 2017

AbstractThe increasing market demands of β‐carotene as colorant, antioxidant and vitamin precursor, requires novel biotechnological production platforms. Yarrowia lipolytica, is an industrial organism unable to naturally synthesize carotenoids but with the ability to produce high amounts of the precursor Acetyl‐CoA. We first found that a lipid overproducer strain was capable of producing more β‐carotene than a wild type after expressing the heterologous pathway. Thereafter, we developed a combinatorial synthetic biology approach base on Golden Gate DNA assembly to screen the optimum promoter‐gene pairs for each transcriptional unit expressed. The best strain reached a production titer of 1.5 g/L and a maximum yield of 0.048 g/g of glucose in flask. β‐carotene production was further increased in controlled conditions using a fed‐batch fermentation. A total production of β‐carotene of 6.5 g/L and 90 mg/g DCW with a concomitant production of 42.6 g/L of lipids was achieved. Such high titers suggest that engineered Y. lipolytica is a competitive producer organism of β‐carotene.
Metabolite profiling of ginsenosides in rat plasma, urine and feces by LC–MS/MS and its application to a pharmacokinetic study after oral administration of Panax ginseng extract
Authors: Dong Wei‐Wei , Han Xiong‐Zhe , Zhao Jinhua , Zhong Fei‐Liang , Ma Rui , Wu Songquan , Li Donghao , Quan Lin‐Hu , Jiang Jun 
Published: October 2017

AbstractPanax ginseng is widely consumed as a functional food in the form of tea, powder, capsules, among others, and possesses a range of pharmacological activities including adaptogenic, immune‐modulatory, anti‐tumor, anti‐aging and anti‐inflammatory effects. The aim of this study was to identify and quantify the major ginsenosides and their metabolites in rat plasma, urine and feces after administration of P. ginseng extract using LC–MS/MS. We collected rat plasma samples at 0.5, 1, 2, 4, 8, 12, 24 and 48 h, and the amounts of urine and fecal samples accumulated in 24 h. Fourteen major ginsenosides and their metabolites were observed in fecal samples at high levels; however, low levels of 11 ginsenosides were detected in urine samples. The pharmacokinetics of the major ginsenosides and their metabolites was investigated in plasma. The results indicated that the maximum plasma concentration, time to maximum concentration and area under the curve of compound K were significantly greater than those of other ginsenosides. This study thus provides valuable information for drug development and clinical application of P. ginseng.
Vision In Stroke cohort: Profile overview of visual impairment
Authors: Rowe Fiona J. ,   the VIS writing Group
Published: October 2017

AbstractAimTo profile the full range of visual disorders from a large prospective observation study of stroke survivors referred by stroke multidisciplinary teams to orthoptic services with suspected visual problems.MethodsMulticenter prospective study undertaken in 20 acute Trust hospitals. Standardized screening/referral forms and investigation forms documented data on referral signs and symptoms plus type and extent of visual impairment.ResultsOf 1,345 patients referred with suspected visual impairment, 915 were recruited (59% men; mean age at stroke onset 69 years [SD 14]). Initial visual assessment was at median 22 days post stroke onset. Eight percent had normal visual assessment. Of 92% with confirmed visual impairment, 24% had reduced central visual acuity <0.3 logMAR and 13.5% <0.5 logMAR. Acquired strabismus was noted in 16% and acquired ocular motility disorders in 68%. Peripheral visual field loss was present in 52%, most commonly homonymous hemianopia. Fifteen percent had visual inattention and 4.6% had other visual perceptual disorders. Overall 84% were visually symptomatic with visual field loss the most common complaint followed by blurred vision, reading difficulty, and diplopia. Treatment options were provided to all with confirmed visual impairment. Targeted advice was most commonly provided along with refraction, prisms, and occlusion.ConclusionsThere are a wide range of visual disorders that occur following stroke and, frequently, with visual symptoms. There are equally a wide variety of treatment options available for these individuals. All stroke survivors require screening for visual impairment and warrant referral for specialist assessment and targeted treatment specific to the type of visual impairment.
Native Chemical Ligation Directed by Photocleavable Peptide Nucleic Acid (PNA) Templates
Authors: Middel Stephen , Panse Cornelia H. , Nawratil Swantje , Diederichsen Ulf 
Published: October 2017

AbstractA novel peptide–peptide ligation strategy is introduced that has the potential to provide peptide libraries of linearly or branched coupled fragments and will be suited to introduce simultaneous protein modifications at different ligation sites. Ligation is assisted by templating peptide nucleic acid (PNA) strands, and therefore, ligation specificity is solely encoded by the PNA sequence. PNA templating, in general, allows for various kinds of covalent ligation reactions. As a proof of principle, a native chemical ligation strategy was elaborated. This PNA‐templated ligation includes easy on‐resin procedures to couple linkers and PNA to the respective peptides, and a traceless photocleavage of the linker/PNA oligomer after the ligation step. A 4,5‐dimethoxy‐2‐nitrobenzaldehyde‐based linker that allowed the photocleavable linkage of two bio‐oligomers was developed.
CRISPR‐Cas9 mediated gene knockout in human coronary artery endothelial cells reveals a pro‐inflammatory role of TLR2
Authors: Wang Yingge , Chen Lu , Tian Zheng , Shen Xueyi , Wang Xiaohong , Wu Honghai , Wang Yayi , Zou Jiayu , Liang Jingyan 
Published: October 2017

AbstractEndothelial inflammatory responses promote the development and progression of atherosclerosis. It was reported that Toll‐like receptors 2 (TLR2) is associated with endothelial inflammation. However, the effect of TLR2 on inflammatory responses in human coronary artery endothelial cells (HCAECs) remains largely unknown. Here, we tested the hypothesis that TLR2 can enhance inflammatory reactions in HCAECs after stimulated by TLR2 agonist. First, we used CRISPR‐Cas9 technology to knockout TLR2 gene in HCAECs. Then, TLR2‐KO and wild type HCAECs were treated with TLR2 agonist peptidoglycan (PGN). The expression levels of intercellular cell adhesion molecule‐1 (ICAM‐1), interleukin‐6 (IL‐6), and interleukin‐8 (IL‐8) were analyzed by real‐time PCR, Western blot, and ELISA. The expression status of myeloid differentiation primary response gene 88 (MyD88), phosphorylated IRAK‐1 (pIRAK‐1) and phosphorylated NF‐κB (pNF‐κB) were detected by Western blot. Our results show that after treated with TLR2 agonist, the expression levels of ICAM‐1, IL‐6, and IL‐8 were downregulated in TLR2‐KO cells compared to those of wild type cells. Further, Western blots of MyD88, pIRAK‐1, and pNF‐κB show that the expression levels of these pro‐inflammatory molecules were much lower in TLR2‐KO cells compared to that of wild type cells by stimulating with TLR2 agonist. We suggest that TLR2 may affect inflammatory reaction in HCAECs by introducing pro‐inflammatory molecules like MyD88, pIRAK‐1, and pNF‐κB.
Brain biopsy in the diagnosis of leptomeningeal involvement in stage I chronic lymphocytic leukemia
Authors: Cervilla Muñoz Eva , Demelo Rodríguez Pablo , García García Alejandra , Menarguez Palanca Javier , del Toro Cervera Jorge 
Published: October 2017

Key Clinical MessageLeptomeningeal involvement of CLL is usually underdiagnosed as neurological symptoms are unspecific. It is important to carefully evaluate neurological status in these patients and consider this entity between the differential diagnosis of a neurological deterioration as adequate treatment improves the prognosis. Imaging techniques, analyses of cerebrospinal fluid, and brain biopsy are useful to establish a definitive diagnosis.
Molecular Viscosity Sensors with Two Rotators for Optimizing the Fluorescence Intensity–Contrast Trade‐Off
Authors: Lee Seung‐Chul , Lee Chang‐Lyoul , Heo Jeongyun , Jeong Chan‐Uk , Lee Gyeong‐Hui , Kim Sehoon , Yoon Woojin , Yun Hoseop , Park Sung O. , Kwak Sang Kyu , Park Sung‐Ha , Kwon O‐Pil 
Published: October 2017

AbstractA series of fluorescent molecular rotors obtained by introducing two rotational groups (“rotators”), which exhibit different rotational and electron‐donating abilities, are discussed. Whereas the control molecular rotor, PH, includes a single rotator (the widely used phenyl group), the PO molecular rotors consist of two rotators (a phenyl group and an alkoxy group), which exhibit simultaneous strongly electron‐donating and easy rotational abilities. Compared with the control rotor PH, PO molecular rotors exhibited one order of magnitude higher quantum yield (fluorescence intensity) and simultaneously exhibited significantly higher fluorescence contrast. These properties are directly related to the strong electron‐donating ability and low energy barrier of rotation of the alkoxy group, as confirmed by dynamic fluorescence experiments and quantum chemical calculations. The PO molecular rotors exhibited two fluorescence relaxation pathways, whereas the PH molecular rotor exhibited a single fluorescence relaxation pathway. Cellular fluorescence imaging with PO molecular rotors for mapping cellular viscosity was successfully demonstrated.
The effective promotion of trace amount of Cu on Ce/WO3‐ZrO2‐TiO2 monolithic catalyst for the low‐temperature NH3‐SCR of NOx
Authors: Li Yuanshan , Xu Haidi , Feng Xi , Liu Shuang , Chen Yaoqiang 
Published: October 2017

AbstractThe selective catalytic reduction of NOx with NH3 (NH3‐SCR) activity of 0.10 g/g CeO2/WO3‐ZrO2‐TiO2 (Ce/WZT) in low temperature range was greatly enhanced by the addition of 0.00010 g/g Cu (Cu/Ce mass ratio = 1:1000). The temperature of complete NOx conversion (T90) decreased as much as 36  °C (from 263  °C to 227  °C) under a gas hourly space velocity (GHSV) of 30 000 h−1. XPS, NH3/O2‐TPD, H2‐TPR, XRD, and FTIR characterizations revealed that the interaction between Cu and Ce, more active oxygen species, stronger NH3 and NO oxidation ability, better redox ability, together with the abundant Brønsted acid sites contributed to better catalytic activity of 0.00010 g/g Cu‐0.10 g/g Ce/WO3‐ZrO2‐TiO2 (CC/WZT) catalyst at low temperatures.