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https://www.readbyqxmd.com/read/28823035/armored-dna-in-recombinant-baculoviruses-as-controls-in-molecular-genetic-assays
#1
Andrea Freystetter, Christian Paar, Herbert Stekel, Jörg Berg
The widespread use of molecular PCR-based assays in analytical and clinical laboratories brings about the need for test-specific, stable, and reliable external controls (EC) as well as standards and internal amplification controls (IC), in order to arrive at consistent test results. In addition, there is also a growing need to produce and provide stable, well-characterized molecular controls for quality assurance programs. In this study, we describe a novel approach to generate armored double-stranded DNA controls, which are encapsulated in baculovirus (BV) particles of the species Autographa californica multiple nucleopolyhedrovirus...
August 19, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28822988/the-components-of-shear-stress-affecting-insect-cells-used-with-the-baculovirus-expression-vector-system
#2
Tobias Weidner, Damir Druzinec, Martina Mühlmann, Rainer Buchholz, Peter Czermak
Insect-based expression platforms such as the baculovirus expression vector system (BEVS) are widely used for the laboratory- and industrial-scale production of recombinant proteins. Thereby, major drawbacks to gain high-quality proteins are the lytic infection cycle and the shear sensitivity of infected insect cells due to turbulence and aeration. Smaller bubbles were formerly assumed to be more harmful than larger ones, but we found that cell damage is also dependent on the concentration of protective agents such as Pluronic®...
August 21, 2017: Zeitschrift Für Naturforschung. C, A Journal of Biosciences
https://www.readbyqxmd.com/read/28822957/two-year-recombinant-human-growth-hormone-rhgh-treatment-is-more-effective-in-pre-pubertal-compared-to-pubertal-short-children-with-x-linked-hypophosphatemic-rickets-xlhr
#3
Anya Rothenbuhler, Laure Esterle, Iva Gueorguieva, Jean-Pierre Salles, Brigitte Mignot, Michel Colle, Agnes Linglart
CONTEXT: Twenty-five to 40% of patients with well-controlled X-linked hypophosphatemic rickets (XLHR) have a final height under -2 SDS. Previous studies have shown that recombinant human growth hormone (rhGH) treatment improves linear growth in short children with XLHR. OBJECTIVE: We studied the effectiveness of rhGH treatment in children with XLHR in a larger cohort. DESIGN: Monocentric, prospective, non-randomized trial. SETTING: University hospital in France...
August 15, 2017: Growth Hormone & IGF Research
https://www.readbyqxmd.com/read/28822825/comparison-of-transforaminal-lumbar-interbody-fusion-outcomes-in-patients-receiving-rhbmp-2-versus-autograft
#4
Taleef R Khan, Kalin R Pearce, Steven J McAnany, Colleen M Peters, Munish C Gupta, Lukas P Zebala
BACKGROUND CONTEXT: Recombinant human bone morphogenetic protein 2 (rhBMP-2) plays a pivotal role in complex spine surgery. Despite its limited approval, the off-label use of rhBMP-2 is prevalent, particularly in transforaminal lumbar interbody fusions (TLIF) PURPOSE: To determine the effectiveness and safety of rhBMP-2 use in TLIF procedures versus autograft. STUDY DESIGN: Retrospective Cohort Study PATIENT SAMPLE: Patients older than 18 years undergoing spine surgery for lumbar degenerative spine disease at a single academic institution...
August 16, 2017: Spine Journal: Official Journal of the North American Spine Society
https://www.readbyqxmd.com/read/28822816/genetically-modified-rabies-virus-vectored-ebola-virus-disease-vaccines-are-safe-and-induce-efficacious-immune-responses-in-mice-and-dogs
#5
Lei Shuai, Xijun Wang, Zhiyuan Wen, Jinying Ge, Jinliang Wang, Dandan Zhao, Zhigao Bu
Ebola viruses (EBOVs) are zoonotic pathogens that cause EBOV disease (EVD) with high case fatality in humans. Currently, EVD vaccines are still under development in several countries. Here, we generated two recombinant rabies viruses (RABVs), rERAG333E/ZGP and rERAG333E/SGP, expressing the Zaire EBOV glycoprotein (ZGP) or Sudan EBOV glycoprotein (SGP) gene based on a modified ERA vaccine strain (rERAG333E) vector platform. The recombinant RABVs retained growth properties similar to those of the vector virus in BSR cell culture and efficiently expressed ZGP or SGP...
August 16, 2017: Antiviral Research
https://www.readbyqxmd.com/read/28822800/amelogenin-induces-m2-macrophage-polarisation-via-pge2-camp-signalling-pathway
#6
Kensuke Yamamichi, Takao Fukuda, Terukazu Sanui, Kyosuke Toyoda, Urara Tanaka, Yuki Nakao, Karen Yotsumoto, Hiroaki Yamato, Takaharu Taketomi, Takeshi Uchiumi, Fusanori Nishimura
OBJECTIVES: Amelogenin, the major component of the enamel matrix derivative (EMD), has been suggested as a bioactive candidate for periodontal regeneration. Apart from producing a regenerative effect on periodontal tissues, amelogenin has also been reported to have an anti-inflammatory effect. However, the precise molecular mechanisms underlying these effects remain unclear. In the present study, we examined the immunomodulatory effects of amelogenin on macrophages. DESIGN: Human phorbol 12-myristate 13-acetate (PMA)-differentiated U937 macrophages and CD14+ peripheral blood-derived monocytes (PBMC)-derived macrophages were stimulated with recombinant amelogenin (rM180)...
August 12, 2017: Archives of Oral Biology
https://www.readbyqxmd.com/read/28822700/full-length-genome-of-a-novel-genotype-3-hepatitis-e-virus-strain-obtained-from-domestic-pigs-in-japan
#7
Putu Prathiwi Primadharsini, Masao Miyake, Satoshi Kunita, Tsutomu Nishizawa, Masaharu Takahashi, Shigeo Nagashima, Tanggis, Hiroshi Ohnishi, Tominari Kobayashi, Takashi Nishiyama, Suljid Jirintai, Hiroaki Okamoto
Hepatitis E virus (HEV) causes acute or chronic hepatitis in humans and can be transmitted via the fecal-oral route. Pigs are one of the main reservoirs for this infection. Sixty pigs, 4-5 months of age, on a swine herd in Japan had detectable anti-HEV IgG antibodies, and five (8.3%) of them had ongoing infection of genotype 3 HEV. Five HEV strains obtained from the viremic pigs shared 98.8-100% nucleotide identity, and one representative strain (swHE1606845), whose entire genomic sequence was determined in this study, differed by 14...
August 16, 2017: Virus Research
https://www.readbyqxmd.com/read/28822642/a-suicidal-strain-of-listeria-monocytogenes-is-effective-as-a-dna-vaccine-delivery-system-for-oral-administration
#8
Shubhra Sinha, Cheng-Yi Kuo, Joan K Ho, Paul J White, Jalal A Jazayeri, Colin W Pouton
In this study we determined the in vivo activity of model ovalbumin vaccines delivered by direct intramuscular delivery of plasmid DNA or oral delivery using a recombinant suicidal Listeria monocytogenes strain (rsΔ2). In a previous report we described how rsΔ2 is capable of delivering luciferase, as protein or DNA, in vitro, into non-dividing intestinal epithelial cells (Kuo et al., 2009). This is achieved by engineering a dual expression shuttle vector, pDuLX-Luc, that replicates in E. coli and rsΔ2 and drives gene expression from the Listeria promoter (Phly) as well as the eukaryotic cytomegalovirus promoter (CMV), thereby delivering both protein and plasmid DNA to the cell cytoplasm...
August 16, 2017: Vaccine
https://www.readbyqxmd.com/read/28822454/-effects-of-up-regulation-of-cathepsin-s-by-hbx-gene-on-hepg2-cell
#9
Z Zhang, P Guo, X Sun, Y Xiao, J Xu, Z S Fang, S Y Wu, H Huang
Objective: To explore the expression of cathepsin S (Cat S) and Hepatitis B virus X protein (HBx) in HBeAg(-) and HBeAg(+ ) hepatocellular carcinoma (HCC), and discuss the effects of Cat S and HBx interaction on HepG2 cell. Methods: Seventy HCC tissue specimens were collected from the surgical resection which were confirmed by pathology in the Fifth Affiliated Hospital of Guangxi Medical University. The tissue samples were separated into two groups: HBeAg(-) group and HBeAg(+ ) group according to the serology of HBeAg...
August 8, 2017: Zhonghua Yi Xue za Zhi [Chinese medical journal]
https://www.readbyqxmd.com/read/28822324/preventive-effects-of-interleukin-6-in-lipopolysaccharide-d-galactosamine-induced-acute-liver-injury-via-regulating-inflammatory-response-in-hepatic-macrophages
#10
Long Li, Chaoli Duan, Yan Zhao, Xiaofang Zhang, Hongyan Yin, Tianxi Wang, Caoxin Huang, Suhuan Liu, Shuyu Yang, Xuejun Li
Lipopolysaccharide/d-Galactosamine (LPS/d-Gal)-induced acute liver injury is characterized by significant inflammatory responses including TNF-α and interleukin-6 (IL-6) and is a widely applied experimental model for inflammation research. TNF-α is critical in the progression of LPS/d-Gal-induced liver injury. However, the role of IL-6 in this model is still unknown. In the present study, we aim to elucidate the involvement of IL-6 in the pathogenesis of acute liver injury induced by LPS/d-Gal in mice and its underlying mechanism...
August 16, 2017: International Immunopharmacology
https://www.readbyqxmd.com/read/28822187/-enzymatic-glycosylation-of-4-demethylepipodophyllotoxin
#11
Ke-Bo Xie, Ri-Dao Chen, Yu-Jiao Zhang, Da-Wei Chen, Jun-Gui Dai
The glycosides of 4'-demethylepipodophyllotoxin (DMEP) possess various pharmacological activities; however, the chemical synthesis of these glycosides faces challenges in regioselectivity, stereoselectivity, and the protection and de-protection of functional groups. In this work, a novel glycosyltransferase (GT) gene AbGT5 from Aloe barbadensis was successfully cloned, heterogeneously expressed and purified. Recombinant AbGT5 was able to catalyze the glycosylation of DMEP and the glycosylated product, which was separated from the preparative scale reaction, was characterized as DMEP 4'-O-β-D-glucoside via MS, 1H-NMR, 13C-NMR, HSQC and HMBC...
June 2017: Zhongguo Zhong Yao za Zhi, Zhongguo Zhongyao Zazhi, China Journal of Chinese Materia Medica
https://www.readbyqxmd.com/read/28822127/in-vitro-analysis-of-metabolite-transport-proteins
#12
Marc-Sven Roell, Franziska Kuhnert, Shirin Zamani-Nour, Andreas P M Weber
The photorespiratory cycle is distributed over four cellular compartments, the chloroplast, peroxisomes, cytoplasm, and mitochondria. Shuttling of photorespiratory intermediates between these compartments is essential to maintain the function of photorespiration. Specific transport proteins mediate the transport across biological membranes and represent important components of the cellular metabolism. Although significant progress was made in the last years on identifying and characterizing new transport proteins, the overall picture of intracellular metabolite transporters is still rather incomplete...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28822125/in-vitro-alkylation-methods-for-assessing-the-protein-redox-state
#13
Flavien Zannini, Jérémy Couturier, Olivier Keech, Nicolas Rouhier
Cysteines are important residues for protein structure, function, and regulation. Owing to their modified reactivity, some cysteines can undergo very diverse redox posttranslational modifications, including the reversible formation of disulfide bonds, a widespread protein regulatory process as well exemplified in plant chloroplasts for Calvin-Benson cycle enzymes. Both core- and peripheral-photorespiratory enzymes possess conserved cysteines, some of which have been identified as being subject to oxidative modifications...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28822070/chemical-shift-assignments-for-the-apo-form-of-the-catalytic-domain-the-linker-region-and-the-carbohydrate-binding-domain-of-the-cellulose-active-lytic-polysaccharide-monooxygenase-sclpmo10c
#14
Gaston Courtade, Zarah Forsberg, Gustav Vaaje-Kolstad, Vincent G H Eijsink, Finn L Aachmann
The apo-form of the 21.4 kDa catalytic domain and the 10.7 kDa carbohydrate binding domain of the AA10 family lytic polysaccharide monooxygenase ScLPMO10C from Streptomyces coelicolor have been isotopically labeled and recombinantly expressed in Escherichia coli. In this paper, we report the (1)H, (13)C, and (15)N chemical shift assignments of each individual domain as well as an ensemble of the assignment for the full-length protein, including its approximately 30-amino acid long linker.
August 18, 2017: Biomolecular NMR Assignments
https://www.readbyqxmd.com/read/28822054/utility-and-comparative-efficacy-of-recombinant-allergens-versus-allergen-extract
#15
REVIEW
Hardik D Patel, Jeffrey M Chambliss, Meera R Gupta
Allergy immunotherapy (AIT) is the only disease-modifying therapy for the treatment of allergic diseases. Although its efficacy and utility are well-established, the potential for serious adverse events, cumbersome and lengthy treatment protocols, and variability of natural allergen preparations have limited its widespread application. Recent advances in recombinant technology have opened new avenues for the development of AIT vaccines. The purpose of this review is to highlight recent evidence on the use of novel recombinant vaccines and review the mechanisms, efficacy, safety, and limitations of AIT...
August 18, 2017: Current Allergy and Asthma Reports
https://www.readbyqxmd.com/read/28822035/identification-and-functional-characterization-of-a-p-coumaroyl-coa-2-hydroxylase-involved-in-the-biosynthesis-of-coumarin-skeleton-from-peucedanum-praeruptorum-dunn
#16
Ruolan Yao, Yucheng Zhao, Tingting Liu, Chuanlong Huang, Sheng Xu, Ziwei Sui, Jun Luo, Lingyi Kong
A p-coumaroyl CoA 2'-hydroxylase responsible for the formation of coumarin lactone ring was identified from Peucedanum praeruptorum Dunn and functionally characterized in vitro. Coumarins are important plant secondary metabolites with a variety of biological activities. Ortho-hydroxylation of cinnamates leads to the formation of coumarin lactone ring and is generally thought to be a key step in coumarin biosynthesis. However, ortho-hydroxylases, especially p-coumaroyl CoA 2'-hydroxylase (C2'H) responsible for the biosynthesis of the most common coumarin skeleton, have received insufficient attention...
August 18, 2017: Plant Molecular Biology
https://www.readbyqxmd.com/read/28822007/brca1-like-profile-is-not-significantly-associated-with-survival-benefit-of-non-myeloablative-intensified-chemotherapy-in-the-gain-randomized-controlled-trial
#17
A G J van Rossum, P C Schouten, K E Weber, V Nekljudova, C Denkert, C Solbach, C H Köhne, C Thomssen, H Forstbauer, G Hoffmann, A Kohls, S Schmatloch, C Schem, G von Minckwitz, T Karn, V J Möbus, S C Linn, S Loibl, F Marmé
PURPOSE: The BRCA1-like profile identifies tumors with a defect in homologous recombination due to inactivation of BRCA1. This profile has been shown to predict which stage III breast cancer patients benefit from myeloablative, DNA double-strand-break-inducing chemotherapy. We tested the predictive potential of the BRCA1-like profile for adjuvant non-myeloablative, intensified dose-dense chemotherapy in the GAIN trial. METHODS: Lymph node positive breast cancer patients were randomized to 3 × 3 dose-dense cycles of intensified epirubicin, paclitaxel, and cyclophosphamide (ETC) or 4 cycles concurrent epirubicin and cyclophosphamide followed by 10 cycles of weekly paclitaxel combined with 4 cycles capecitabine (EC-TX)...
August 18, 2017: Breast Cancer Research and Treatment
https://www.readbyqxmd.com/read/28821968/near-adult-height-in-male-kidney-transplant-recipients-started-on-growth-hormone-treatment-in-late-puberty
#18
Silvia Gil, Mariana Aziz, Marta Adragna, Marta Monteverde, Alicia Belgorosky
BACKGROUND: Growth retardation and its impact on adult height is considered to be one of the most common complications in patients with chronic kidney disease (CKD). Treatment with recombinant human growth hormone (rhGH) has been effective in improving growth in kidney transplantation (KTx) patients, but little data are available on adult height in patients who began rhGh treatment in late puberty. METHODS: Near-adult height was evaluated in 13 KTx patients treated with rhGH [growth hormone group (GHGr); dose 9...
August 18, 2017: Pediatric Nephrology: Journal of the International Pediatric Nephrology Association
https://www.readbyqxmd.com/read/28821946/quantitative-expression-analysis-of-spa-fnba-and-rsp-genes-in-staphylococcus-aureus-actively-associated-in-the-formation-of-biofilms
#19
Sthanikam Yeswanth, Abhijit Chaudhury, Potukuchi Venkata Gurunadha Krishna Sarma
In Staphylococcus aureus, adherence and secretory proteins play chief role in the formation of biofilms. This mode of growth exhibits resistance to a variety of antibiotics and spreads its infections. In the present study, secretary and adherence proteins, Protein-A, Fibronectin-binding protein-A (FnbA) and Rsp (a transcription regulator encoding proteolytic property) expression levels were evaluated at different stages of growth in S. aureus ATCC12600 a drug-sensitive strain and multidrug-resistant strains of S...
August 18, 2017: Current Microbiology
https://www.readbyqxmd.com/read/28821784/high-copy-and-stable-expression-of-the-xylanase-xynhb-in-saccharomyces-cerevisiae-by-rdna-mediated-integration
#20
Cheng Fang, Qinhong Wang, Jonathan Nimal Selvaraj, Yuling Zhou, Lixin Ma, Guimin Zhang, Yanhe Ma
Xylanase is a widely-used additive in baking industry for enhancing dough and bread quality. Several xylanases used in baking industry were expressed in different systems, but their expression in antibiotic free vector system is highly essential and safe. In the present study, an alternative rDNA-mediated technology was developed to increase the copy number of target gene by integrating it into Saccharomyces cerevisiae genome. A xylanase-encoding gene xynHB from Bacillus sp. was cloned into pHBM367H and integrated into S...
August 18, 2017: Scientific Reports
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