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https://www.readbyqxmd.com/read/28216260/optimal-expression-of-a-fab-effector-fusion-protein-in-escherichia-coli-by-removing-the-cysteine-residues-responsible-for-an-interchain-disulfide-bond-of-a-fab-molecule
#1
Hyeon-Ju Kang, Hye-Jin Kim, Mun-Sik Jung, Jae-Kyu Han, Sang-Hoon Cha
Development of novel bi-functional or even tri-functional Fab-effector fusion proteins would have a great potential in the biomedical sciences. However, the expression of Fab-effector fusion proteins in Escherichia coli is problematic especially when a eukaryotic effector moiety is genetically linked to a Fab due to the lack of proper chaperone proteins and an inappropriate physicochemical environment intrinsic to the microbial hosts. We previously reported that a human Fab molecule, referred to as SL335, reactive to human serum albumin has a prolonged in vivo serum half-life in rats...
February 16, 2017: Immunology Letters
https://www.readbyqxmd.com/read/28131652/precise-and-combinatorial-pegylation-generates-a-low-immunogenic-and-stable-form-of-human-growth-hormone
#2
Ling Wu, Jingxian Chen, Yiming Wu, Bo Zhang, Xiaochun Cai, Ziwei Zhang, Yan Wang, LongLong Si, Huan Xu, YongXiang Zheng, ChuanLing Zhang, Chenggang Liang, Jing Li, Lihe Zhang, Qiang Zhang, Demin Zhou
In this study, we aimed to develop a safe and stable form of human growth hormone (hGH) and to refine PEGylation methods for therapeutic proteins via genetic code expansion. Through this precise approach, a series of polyethylene glycol (PEG) moieties and sites were combined in various ways. Additionally, the effects of combinatorial PEGylation on the biological, pharmacological, and immunogenic properties of hGH in vitro and vivo were analyzed. Our results showed that combinatorial PEGylation at Y35, G131, and K145 significantly reduced immunogenicity and improved pharmacokinetic (PK) profiles compared with mono-PEGylation, while retaining biological activity...
January 25, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28105973/selective-determination-of-human-growth-hormone-somatropin-in-the-presence-of-its-chemical-degradation-products
#3
Mamdouh R Rezk, Marwa F Mohamed, Faten Abdelaziz Fathalla, Mostafa A Shehata
A rapid and sensitive HPLC method was developedand validated for selective determination of thehuman growth hormone (hGH) somatropin in thepresence of its deamidated and oxidized degradationproducts. Reversed-phase chromatographywith an acetonitrile and ammonium bicarbonatemobile phase in gradient elution mode was used.A short run time of 15 min allowed rapid andcost-effective analysis, with an average retentiontime of 7.4 min for native hGH, 6.2 min for itsdeamidated form, and 4.3 and 6 min for its oxidizedvariants...
January 19, 2017: Journal of AOAC International
https://www.readbyqxmd.com/read/28099130/epidermal-growth-factor-receptor-egfr-involvement-in-successful-growth-hormone-gh-signaling-in-gh-transduction-defect
#4
Eirini Kostopoulou, Andrea Paola Rojas-Gil, Alexia Karvela, Bessie E Spiliotis
BACKGROUND: Growth hormone (GH) transduction defect (GHTD) is a growth disorder with impaired signal transducer and activator of transcription 3 (STAT3) phosphorylation mediated by overexpression of cytokine-inducible SH2-containing protein (CIS), which causes increased growth hormone receptor (GHR) degradation. This study investigated the role of epidermal growth factor (EGF) in the restoration of normal GH signaling in GHTD. METHODS: Protein expression, cellular localization and physical contact of proteins of the GH and EGF signaling pathways were studied by Western immunoblotting, immunofluorescence and co-immunoprecipitation, respectively...
February 1, 2017: Journal of Pediatric Endocrinology & Metabolism: JPEM
https://www.readbyqxmd.com/read/28013084/expression-purification-and-characterization-of-the-authentic-form-of-human-growth-hormone-receptor-antagonist-g120r-hgh-obtained-in-escherichia-coli-periplasmic-space
#5
Ana C S C Menezes, Miriam F Suzuki, João E Oliveira, Maria T C P Ribela, Isadora C Furigo, José Donato, Paolo Bartolini, Carlos R J Soares
The human growth hormone receptor antagonist G120R-hGH precludes dimerization of GH and prolactin receptors and consequently JAK/STAT signaling. Some modifications in this antagonist resulted in a drug specific for the GH receptor, called Pegvisomant (Somavert(®)). However, the original G120R-hGH is usually synthesized in bacterial cytoplasm as inclusion bodies, not being a commercial product. The present work describes the synthesis and characterization of G120R-hGH secreted into bacterial periplasm and obtained with a vector based on a constitutive lambda-PL promoter...
December 22, 2016: Protein Expression and Purification
https://www.readbyqxmd.com/read/28003839/twin-arginine-translocation-system-in-secretory-expression-of-recombinant-human-growth-hormone
#6
Mohammad Reza Bagherinejad, Hamid Mir-Mohammad Sadeghi, Daryoush Abedi, C Perry Chou, Fatemeh Moazen, Mohammad Rabbani
Recombinant protein production in E. coli has several advantages over other expression systems. Misfolding, inclusion body formation, and lack of eukaryotic post translational modification are the most disadvantages of this system. Exporting of correctly folded proteins to the outside of reductive cytoplasmic environment through twin-arginine system could help to pass these limiting steps. Two signal sequences, TorA and SufI are used at N-terminal of human growth hormone (hGH) bearing DsbA gene sequence at C-terminal to enhance folding...
December 2016: Research in Pharmaceutical Sciences
https://www.readbyqxmd.com/read/27973772/relativistic-effects-on-electron-nucleus-hyperfine-coupling-studied-with-an-exact-2-component-x2c-hamiltonian
#7
Jochen Autschbach
An exact 2-component (X2C) transformation of the one-electron Hamiltonian is used to transform nuclear hyperfine magnetic field operators from the 4-component Dirac picture to 2-component form. Numerical applications are concerned with hyperfine coupling constants of one-electron and many-electron atoms, as well as the HgH radical, using spin-unrestricted scalar X2C Hartree-Fock and Kohn-Sham theory. Reference data for 2-component generalized-collinear X2C calculations, including spin-orbit coupling, are also provided for selected cases...
January 3, 2017: Journal of Chemical Theory and Computation
https://www.readbyqxmd.com/read/27942455/the-age-dependent-effects-of-a-single-dose-methylphenidate-challenge-on-cerebral-perfusion-in-patients-with-attention-deficit-hyperactivity-disorder
#8
A Schrantee, Hjmm Mutsaerts, C Bouziane, Hgh Tamminga, M A Bottelier, L Reneman
Methylphenidate (MPH) is a stimulant drug and an effective treatment for attention-deficit/hyperactivity disorder (ADHD) in both children and adults. Pre-clinical studies suggest that the response to stimulants is dependent on age, which may reflect the ontogeny of the dopamine (DA) system, which continues to develop throughout childhood and adolescence. Therefore, the aim of this study was to investigate the modulating effect of age on the cerebral blood flow (CBF) response to MPH in stimulant treatment-naive children and adults with ADHD...
2017: NeuroImage: Clinical
https://www.readbyqxmd.com/read/27932411/mod-4023-a-long-acting-carboxy-terminal-peptide-modified-human-growth-hormone-results-of-a-phase-2-study-in-growth-hormone-deficient-adults
#9
RANDOMIZED CONTROLLED TRIAL
Christian J Strasburger, Peter Vanuga, Juraj Payer, Marija Pfeifer, Vera Popovic, László Bajnok, Miklós Góth, Veˇra Olšovská, L'udmila Trejbalová, Janos Vadasz, Eyal Fima, Ronit Koren, Leanne Amitzi, Martin Bidlingmaier, Oren Hershkovitz, Gili Hart, Beverly M K Biller
OBJECTIVE: Growth hormone (GH) replacement therapy currently requires daily injections, which may cause distress and low compliance. C-terminal peptide (CTP)-modified growth hormone (MOD-4023) is being developed as a once-weekly dosing regimen in patients with GH deficiency (GHD). This study's objective is to evaluate the safety, pharmacokinetics (PK), pharmacodynamics (PD) and efficacy of MOD-4023 administered once-weekly in GHD adults. DESIGN: 54 adults with GHD currently treated with daily GH were normalized and randomized into 4 weekly dosing cohorts of MOD-4023 at 18...
March 2017: European Journal of Endocrinology
https://www.readbyqxmd.com/read/27923197/single-chain-fc-dimer-human-growth-hormone-fusion-protein-for-improved-drug-delivery
#10
Li Zhou, Hsuan-Yao Wang, Shanshan Tong, Curtis T Okamoto, Wei-Chiang Shen, Jennica L Zaro
Fc fusion protein technology has been successfully used to generate long-acting forms of several protein therapeutics. In this study, a novel Fc-based drug carrier, single chain Fc-dimer (sc(Fc)2), was designed to contain two Fc domains recombinantly linked via a flexible linker. Since the Fc dimeric structure is maintained through the flexible linker, the hinge region was omitted to further stabilize it against proteolysis and reduce FcγR-related effector functions. The resultant sc(Fc)2 candidate preserved the neonatal Fc receptor (FcRn) binding...
February 2017: Biomaterials
https://www.readbyqxmd.com/read/27920886/evaluation-of-recombinant-human-growth-hormone-secretion-in-e-coli-using-the-l-asparaginase-ii-signal-peptide
#11
Mozhdeh Zamani, Navid Nezafat, Younes Ghasemi
BACKGROUND: In the recent years, there has been an increasing interest in secretory production of recombinant proteins, due to its various advantages compared with intracellular expression. Signal peptides play a critical role in prosperous secretion of recombinant proteins. Accordingly, different signal peptides have been assessed for their ability to produce secretory proteins by trial-and-error experiments. The aim of this study was to evaluate the effect of L-asparaginase II signal peptide on the recombinant human Growth Hormone (hGH) protein secretion in the Escherichia coli (E...
October 2016: Avicenna Journal of Medical Biotechnology
https://www.readbyqxmd.com/read/27857836/refractory-hypoglycemia-in-a-patient-with-functional-adrenal-cortical-carcinoma
#12
Katia Regina Marchetti, Maria Adelaide Albergaria Pereira, Arnaldo Lichtenstein, Edison Ferreira Paiva
: Adrenacarcinomas are rare, and hypoglycemic syndrome resulting from the secretion of insulin-like growth factor II (IGF-II) by these tumors have been described infrequently. This study describes the case of a young woman with severe persistent hypoglycemia and a large adrenal tumor and discusses the physiopathological mechanisms involved in hypoglycemia. The case is described as a 21-year-old woman who presented with 8 months of general symptoms and, in the preceding 3 months, with episodes of mental confusion and visual blurring secondary to hypoglycemia...
2016: Endocrinology, Diabetes & Metabolism Case Reports
https://www.readbyqxmd.com/read/27812793/uk-iatrogenic-creutzfeldt-jakob-disease-investigating-human-prion-transmission-across-genotypic-barriers-using-human-tissue-based-and-molecular-approaches
#13
Diane L Ritchie, Marcelo A Barria, Alexander H Peden, Helen M Yull, James Kirkpatrick, Peter Adlard, James W Ironside, Mark W Head
Creutzfeldt-Jakob disease (CJD) is the prototypic human prion disease that occurs most commonly in sporadic and genetic forms, but it is also transmissible and can be acquired through medical procedures, resulting in iatrogenic CJD (iCJD). The largest numbers of iCJD cases that have occurred worldwide have resulted from contaminated cadaveric pituitary-derived human growth hormone (hGH) and its use to treat primary and secondary growth hormone deficiency. We report a comprehensive, tissue-based and molecular genetic analysis of the largest series of UK hGH-iCJD cases reported to date, including in vitro kinetic molecular modelling of genotypic factors influencing prion transmission...
November 3, 2016: Acta Neuropathologica
https://www.readbyqxmd.com/read/27765563/predictors-of-aggressive-clinical-phenotype-among-immunohistochemically-confirmed-atypical-adenomas
#14
Hasan A Zaidi, David J Cote, Ian F Dunn, Edward R Laws
Despite formal pathological criteria, not all atypical pituitary adenomas display clinically aggressive behavior. We set out to determine which factors predict a clinically aggressive phenotype among a cohort of atypical pituitary adenomas. Medical records were retrospectively reviewed from April 2008 to July 2015. Of 569 pituitary adenomas, 47 (8.3%) patients were surgically treated for atypical adenomas as defined by the WHO criteria. Clinically aggressive adenomas were defined as occurring in those patients who necessitated additional therapeutic intervention after the index (first) surgery, including additional surgery, medical therapy, or radiosurgery...
December 2016: Journal of Clinical Neuroscience: Official Journal of the Neurosurgical Society of Australasia
https://www.readbyqxmd.com/read/27713394/effect-of-growth-hormone-and-igg-aggregates-on-dendritic-cells-activation-and-t-cells-polarization
#15
Yann Gallais, Natacha Szely, François-Xavier Legrand, Arnaud Leroy, Marc Pallardy, Isabelle Turbica
Patients treated with therapeutic biological products (BP) frequently develop anti-drug antibodies (ADA) with potential neutralizing capacities leading to loss of clinical response or serious side effects. BP aggregates have been suggested to promote immunogenicity, thus enhancing ADA production. Dendritic cells (DC) are key effectors in T-cell and B-cell fates, and the subsequent generation of immunogenicity. The objective of this work was to determine if BP aggregates can participate to DC maturation and T-cell activation...
November 8, 2016: Immunology and Cell Biology
https://www.readbyqxmd.com/read/27681688/dysregulation-of-calcium-channels-decreases-parasecretion-in-pancreatic-%C3%AE-cells-in-rats-born-small-for-gestational-age
#16
Yan-Yan Jin, Meng-Zao He, Zhen-Yong Wu, Ke Huang, Ying Shen, Li Liang, Jian-Hua Mao
OBJECTIVE: To investigate the role of intrauterine malnourishment in the development and function of pancreatic islet β-cells. METHODS: Whole-cell patch clamping was used to record voltage-gated calcium channel (VGCC)-mediated currents. Insulin secretion was detected by measuring capacitance using a sequence of sine wave stimuli. VGCC currents and insulin secretion were measured in the small for gestational age (SGA) group treated with human recombinant growth hormone (hGH)...
September 29, 2016: Growth Factors
https://www.readbyqxmd.com/read/27677452/comparative-expression-and-regulation-of-duplicated-fibroblast-growth-factor-1-genes-in-grass-carp-ctenopharyngodon-idella
#17
Dan-Dan Guo, Wen-Zhi Guan, Yi-Wen Sun, Jie Chen, Xia-Yun Jiang, Shu-Ming Zou
Fibroblast growth factor 1 (Fgf1) is known as a mitogenic factor involved in the regulation of cell growth, proliferation, and differentiation in vertebrates. Here, we report the isolation and characterization of two fgf1 genes in grass carp (Ctenopharyngodon idella). Grass carp fgf1a and fgf1b cDNAs are highly divergent, sharing a relatively low amino acid sequence identity of 50%, probably due to fish-specific gene duplication. fgf1a and fgf1b mRNAs were detected in the zygote and expressed throughout embryogenesis...
January 1, 2017: General and Comparative Endocrinology
https://www.readbyqxmd.com/read/27670405/unexpected-effects-of-the-mip-creer-transgene-and-tamoxifen-on-%C3%AE-cell-growth-in-c57bl6-j-male-mice
#18
Bethany A Carboneau, Thao D V Le, Jennifer C Dunn, Maureen Gannon
Transgenic mouse models have been fundamental in the discovery of factors that regulate β-cell development, mass, and function. Several groups have recently shown that some of these models display previously uncharacterized phenotypes due to the transgenic system itself. These include impaired islet function and increased β-cell mass due to the presence of a human growth hormone (hGH) minigene as well as impaired β-cell proliferation in response to tamoxifen (TM) administration. We aimed to determine how these systems impact β-cell mass and proliferation during high fat diet (HFD)...
September 2016: Physiological Reports
https://www.readbyqxmd.com/read/27669684/direct-and-label-free-detection-of-the-human-growth-hormone-in-urine-by-an-ultrasensitive-bimodal-waveguide-biosensor
#19
Ana Belén González-Guerrero, Jesús Maldonado, Stefania Dante, Daniel Grajales, Laura M Lechuga
A label-free interferometric transducer showing a theoretical detection limit for homogeneous sensing of 5 × 10(-8) RIU, being equivalent to a protein mass coverage resolution of 2.8 fg mm(-2) , is used to develop a high sensitive biosensor for protein detection. The extreme sensitivity of this transducer combined with a selective bioreceptor layer enables the direct evaluation of the human growth hormone (hGH) in undiluted urine matrix in the 10 pg mL(-1) range.
January 2017: Journal of Biophotonics
https://www.readbyqxmd.com/read/27665368/preparation-of-soluble-isotopically-labeled-human-growth-hormone-produced-in-escherichia-coli
#20
Jin-Hee Lee, Ji-Seon Jeong, Sook-Kyung Kim, Jimyeong Song, Ji Youn Lee, Soyun Baek, Jun-Hyuk Choi
Isotopically labeled proteins have been used as internal standards for mass spectrometry (MS)-based absolute protein quantification. Although this approach can provide highly accurate analyses of proteins of interest within a complex mixture, one of the major limitations of this method is the difficulty in preparing uniformly labeled standards. Human growth hormone (hGH) is one of the most important hormones that circulate throughout the body, and its measurement is primarily of interest in the diagnosis and treatment of growth disorders...
November 1, 2016: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
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