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https://www.readbyqxmd.com/read/28902870/reduced%C3%A2-insulin-signaling-maintains-electrical-transmission-in-a-neural-circuit-in-aging-flies
#1
Hrvoje Augustin, Kieran McGourty, Marcus J Allen, Sirisha Kudumala Madem, Jennifer Adcott, Fiona Kerr, Chi Tung Wong, Alec Vincent, Tanja Godenschwege, Emmanuel Boucrot, Linda Partridge
Lowered insulin/insulin-like growth factor (IGF) signaling (IIS) can extend healthy lifespan in worms, flies, and mice, but it can also have adverse effects (the "insulin paradox"). Chronic, moderately lowered IIS rescues age-related decline in neurotransmission through the Drosophila giant fiber system (GFS), a simple escape response neuronal circuit, by increasing targeting of the gap junctional protein innexin shaking-B to gap junctions (GJs). Endosomal recycling of GJs was also stimulated in cultured human cells when IIS was reduced...
September 2017: PLoS Biology
https://www.readbyqxmd.com/read/28898422/dynamic-gastric-digestion-of-a-commercial-whey-protein-concentrate
#2
Beatriz Miralles, Roberto Del Barrio, Carolina Cueva, Isidra Recio, Lourdes Amigo
BACKGROUND: A dynamic gastrointestinal simulator, simgi(®) , has been applied to assess the gastric digestion of a whey protein concentrate. Samples collected from the outlet of the stomach have been compared to those resulting from the static digestion protocol INFOGEST developed on the basis of physiologically inferred conditions. RESULTS: Progress of digestion was followed by SDS-PAGE and LC-MS/MS. By SDS-PAGE, serum albumin and α-lactalbumin were no longer detectable at 30 and 60 min, respectively...
September 12, 2017: Journal of the Science of Food and Agriculture
https://www.readbyqxmd.com/read/28887386/fragile-x-mental-retardation-protein-restricts-small-dye-iontophoresis-entry-into-central-neurons
#3
Tyler Kennedy, Kendal Broadie
Fragile X Mental Retardation Protein (FMRP) loss causes Fragile X syndrome (FXS), a major disorder characterized by autism, intellectual disability, hyperactivity and seizures. FMRP is both an RNA- and channel-binding regulator, with critical roles in neural circuit formation and function. However, it remains unclear how these FMRP activities relate to each other and how dysfunction in their absence underlies FXS neurological symptoms. In testing circuit level defects in the Drosophila FXS model, we discovered a completely unexpected and highly robust neuronal dye iontophoresis phenotype in the well-mapped Giant Fiber (GF) circuit...
September 8, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28884428/in-vitro-and-in-vivo-anti-proliferative-evaluation-of-bis-4-4-tolyl-2-2-6-2%C3%A2-terpyridine-copper-ii-complex-against-ehrlich-ascites-carcinoma-tumors
#4
Dharmasivam Mahendiran, Raju Senthil Kumar, Vijayan Viswanathan, Devadasan Velmurugan, Aziz Kalilur Rahiman
The bis(4'-(4-tolyl)-2,2':6',2″-terpyridine)copper(II) complex [Cu(ttpy)2]Cl2 was synthesized and authenticated by single crystal analysis, which shows distorted octahedral geometry around copper(II) ion. The crystal packing is stabilized by C-H···π inter and intramolecular interactions. The complex was found to be lipophilic as determined by shake-flask method. In vitro cytotoxicity of the complex was tested against Ehrlich ascites carcinoma (EAC) and L6 myotube cell lines. The complex exhibit potent cytotoxicity with respect to the commercially available anticancer drug cisplatin...
September 7, 2017: Journal of Biological Inorganic Chemistry: JBIC
https://www.readbyqxmd.com/read/28873703/oil-in-water-emulsions-stabilized-by-tyrosinase-crosslinked-potato-protein
#5
Jovana Glusac, Sivan Isaschar-Ovdat, Biljana Kukavica, Ayelet Fishman
Potato protein (PP) holds great promise as a non-allergenic food ingredient with high nutritional value. Attempts to modulate its functional properties by crosslinking have not been reported to date. The effect of tyrosinase-mediated crosslinking of PP on the properties of o/w emulsions was studied in the present work. Among the various PPs, protease inhibitors were efficiently crosslinked by the enzyme as determined by SDS-PAGE analysis. Concentrated emulsions comprising 40% olive oil and 6.1% PP (w/w) were fabricated by a shear stress homogenizer...
October 2017: Food Research International
https://www.readbyqxmd.com/read/28869737/-a-method-to-increase-the-production-of-insulin-precursor-in-pichia-pastoris
#6
Chenchen Liang, Li Wang, Qiuling Luo, Jing Wu
To improve the yield of insulin precursor (PI), we constructed a recombinant expression vector pPIC9K-PI and transformed it into Pichia pastoris GS115 using electroporation. After screening, a mutant strain CL012 with 12 copies was obtained on the YPDS plate containing 4.0 mg/mL G418. Then, the components of SNAREs (Soluble N-ethylmaleimide-sensitive factor attachment receptor proteins), SNC2 and SNC2-SSO2, were expressed in the strain CL012 to explore the effect of SNAREs on the yield of PI. In shake flask culture, the strains expressing SNC2 and SNC2-SSO2 yielded PI of 1...
July 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/28865117/development-of-a-general-defined-medium-for-pichia-pastoris
#7
Catherine Bartlett Matthews, Angel Kuo, Kerry Routenberg Love, J Christopher Love
Pichia pastoris is widely used as a host for recombinant protein production. More than 500 proteins have been expressed in the organism at a variety of cultivation scales, from small shake flasks to large bioreactors. Large-scale fermentation strategies typically employ chemically-defined growth medium because of its greater batch-to-batch consistency and in many cases, lower costs compared to complex medium. For biopharmaceuticals, defined growth medium may also simplify downstream purification and regulatory documentation...
September 2, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28861662/extracellular-expression-in-aspergillus-niger-of-an-antibody-fused-to-leishmania-sp-antigens
#8
Denis Magaña-Ortíz, Francisco Fernández, Achim M Loske, Miguel A Gómez-Lim
Nucleoside hydrolase and sterol 24-c-methyltransferase, two antigenic proteins of Leishmania sp., were expressed in Aspergillus niger. Genetic transformation of conidia was achieved using underwater shock waves. scFv antibody addressed to DEC205, a receptor of dendritic cells, was fused to two proteins of Leishmania sp. Receptor 205 has a relevant role in the immune system in mammals; it can modulate T cell response to different antigens. Extracellular expression strategy of recombinant antibody was achieved using a fragment of native glucoamylase A (514 aa) as a carrier...
August 31, 2017: Current Microbiology
https://www.readbyqxmd.com/read/28861059/characterization-of-full-length-and-truncated-recombinant-%C3%AE%C2%BA-carrageenase-expressed-in-pichia-pastoris
#9
Yuan Yu, Zhemin Liu, Min Yang, Meng Chen, Zhihan Wei, Lixia Shi, Li Li, Haijin Mou
κ-Carrageenase belongs to glycoside hydrolase family 16 and cleaves the β-(1→4) linkages of κ-carrageenan. In this study, genes encoding the full-length (cgkZ), Por secretion tail-truncated (cgkZΔPst) and carbohydrate binding domain-truncated (cgkZΔCBM) κ-carrageenase proteins were expressed in Pichia pastoris. The copy numbers of gene cgkZ, cgkZΔPst and cgkZΔCBM were 7, 7 and 6, respectively. The enzymatic activities of recombinant enzymes cgkZ, cgkZΔPst and cgkZΔCBM reached 4.68, 5.70, and 3.02 U/mL, respectively, after 120 h of shake flask fermentation at 22°C and pH 6 in the presence of 1 % (v/v) methanol...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28860027/preparation-of-wool-follicles-for-proteomic-studies
#10
Jeffrey E Plowman, Joy L Woods, Bede van Schaijik, Duane P Harland
A variety of techniques were applied to wool follicles stored in William's E culture medium to optimise the extraction of keratin and keratin associated proteins (KAPs). A time course study indicated that the maximum storage time for live skin in this buffer at 20 °C was 24 h, after which degradative loss of protein became significant. Maceration of the skin for 10 min followed by reciprocal action shaking for 14 h had a detrimental effect on keratin extractability. The best approach involved using a Dounce homogeniser as this resulted in the highest amount of Type I and II keratins and KAPs...
August 28, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28842980/escherichia-coli-tatexpress-strains-super-secrete-human-growth-hormone-into-the-bacterial-periplasm-by-the-tat-pathway
#11
Douglas F Browning, Kirsty L Richards, Amber R Peswani, Jo Roobol, Stephen J W Busby, Colin Robinson
Numerous high-value proteins are secreted into the Escherichia coli periplasm by the General Secretory (Sec) pathway, but Sec-based production chassis cannot handle many potential target proteins. The Tat pathway offers a promising alternative because it transports fully folded proteins; however, yields have been too low for commercial use. To facilitate Tat export, we have engineered the TatExpress series of super-secreting strains by introducing the strong inducible bacterial promoter, ptac, upstream of the chromosomal tatABCD operon, to drive its expression in E...
August 26, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28840658/experimental-design-and-metabolic-flux-analysis-tools-to-optimize-industrially-relevant-haemophilus-influenzae-type-b-growth-medium
#12
Mateus Ribeiro da Silva, Carla Andreia Freixo Portela, Silvia Maria Ferreira Albani, Paola Rizzo de Paiva, Martha Massako Tanizaki, Teresa Cristina Zangirolami
Haemophilus influenzae type b (Hib), a Gram-negative capsulated bacterium, is a causative agent of meningitis worldwide. The capsular polysaccharide, a high molecular mass polymer consisting of the repeated units of the polyribosyl-ribitol-phosphate, is considered the main virulence factor and it is used as an antigen to vaccines, conjugated to a carrier protein. The industrial production of the polysaccharide requires the cultivation of Hib in rich medium, which impacts process costs and product recovery. In this study, a central composite rotational experimental design strategy was used to access the influence of key components of culture medium (soy peptone, yeast extract and glucose) on biomass formation and polysaccharide production in shake-flasks...
August 25, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28837572/efficient-and-stable-production-of-modified-vaccinia-ankara-virus-in-two-stage-semi-continuous-and-in-continuous-stirred-tank-cultivation-systems
#13
Felipe Tapia, Ingo Jordan, Yvonne Genzel, Udo Reichl
One important aim in cell culture-based viral vaccine and vector production is the implementation of continuous processes. Such a development has the potential to reduce costs of vaccine manufacturing as volumetric productivity is increased and the manufacturing footprint is reduced. In this work, continuous production of Modified Vaccinia Ankara (MVA) virus was investigated. First, a semi-continuous two-stage cultivation system consisting of two shaker flasks in series was established as a small-scale approach...
2017: PloS One
https://www.readbyqxmd.com/read/28833557/enhancing-recombinant-interleukin-6-production-yield-by-fermentation-optimization-two-steps-denaturing-2dr-and-one-step-purification
#14
Nadeem Ahmed, Rabbia Abbas, Mohsin Ahmad Khan, Hamid Bashir, Saad Tahir, Ahmad Usman Zafar
Interleukin-6 a pleiotropic cytokine involved in wide range biological activities. So the large scale Production of biologically active recombinant human interleukin-6 is important for its structural and functional studies. Here we report an optimized method for shake flask fermentation and simplified high yield purification procedure for the recombinant interleukin -6. This high yield expression method not only involves the optimization of the fermentation condition, but also the single step purification method as well as two-steps denaturing and one step refolding process (2DR)...
August 22, 2017: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/28826344/scalable-lentiviral-vector-production-using-stable-hek293sf-producer-cell-lines
#15
Aziza P Manceur, Howard Kim, Vanja Misic, Nadejda Andreev, July Dorion-Thibaudeau, Stéphane Lanthier, Alice Bernier, Sonia Tremblay, Anne-Marie Gélinas, Sophie Broussau, Rénald Gilbert, Sven Ansorge
Lentiviral vectors (LV) represent a key tool for gene and cell therapy applications. The production of these vectors in sufficient quantities for clinical applications remains a hurdle, prompting the field towards developing suspension processes that are conducive to large scale production. We describe here a LV production strategy using a stable inducible producer cell line. The HEK293 cell line employed grows in suspension, thus offering direct scalability, and produces a Green Fluorescent Protein (GFP)-expressing lentiviral vector in the 10<sup>6</sup> Transduction Units (TU)/ml range without optimization...
August 21, 2017: Human Gene Therapy Methods
https://www.readbyqxmd.com/read/28817974/applications-of-the-real-time-quaking-induced-conversion-assay-in-diagnosis-prion-strain-typing-drug-pre-screening-and-other-amyloidopathies
#16
Niccolò Candelise, Matthias Schmitz, Susana Margarida Da Silva Correia, Amandeep Singh Arora, Anna Villar-Piqué, Saima Zafar, Franc Llorens, Maria Cramm, Inga Zerr
The development of in vitro protein misfolding amplification assays for the detection and analysis of abnormally folded proteins, such as proteinase K resistant prion protein (PrP(res)) was a major innovation in the prion field. In prion diseases, these types of assays imitate the pathological conversion of the cellular PrP (PrP(C)) into a proteinase resistant associated conformer or amyloid, called PrP(res). Areas covered: The most prominent protein misfolding amplification assays are the protein misfolding cyclic amplification (PMCA), which is based on sonication and the real-time quaking-induced conversion (RT-QuIC) technique based on shaking...
September 8, 2017: Expert Review of Molecular Diagnostics
https://www.readbyqxmd.com/read/28802025/simple-and-rapid-determination-of-zaltoprofen-in-human-plasma-by-manual-shaking-assisted-dispersive-liquid-liquid-microextraction-followed-by-liquid-chromatography-with-ultraviolet-detection
#17
Se Young Han, Kyung Min Jeong, Da Eun Yoo, Yan Jin, Eun Mi Kim, Su Hyun Kim, Seok-Yong Lee, Se-Hyung Kim, Jing Zhao, Jeongmi Lee
A readily applicable method was developed to determine the concentration level of zaltoprofen, a non-steroidal anti-inflammatory drug from the propionic acid family, in human plasma. This method is based on manual shaking-assisted dispersive liquid-liquid microextraction coupled with liquid chromatography with UV detection. Factors affecting the extraction efficiency were screened and optimized by experimental design using fractional factorial and central composite designs, respectively. Optimal conditions were: 220 μL of C2 H4 Cl2 (extraction solvent), 5 mL of 3...
August 12, 2017: Journal of Separation Science
https://www.readbyqxmd.com/read/28800398/a-13-week-randomized-pilot-trial-of-a-low-glycemic-load-diet-and-lifestyle-modification-program-combining-low-glycemic-load-protein-shakes-and-targeted-nutraceuticals-improved-weight-loss-and-cardio-metabolic-risk-factors-over-a-low-glycemic-load-diet-alone
#18
Clinton J Dahlberg, Joseph J Ou, John G Babish, Joseph J Lamb, Sarah Eliason, Holly Brabazon, Wei Gao, Mohan R Kaadige, Matthew L Tripp
An open label, randomized, exploratory study comprising 44 healthy, overweight subjects with cardio-metabolic syndrome (CMS) risk factors was conducted to assess the safety, tolerability and efficacy of a proprietary lifestyle modification program without (DIET) and with (PROG) targeted nutraceutical supplementation including soy, pea and whey proteins, phytosterols, antioxidants, probiotics, fish oil and berberine over 13-weeks. Key metrics were recorded at baseline, weeks 9 and 13. For DIET and PROG compliance was 85% and 86%, respectively, with no adverse events related to the diet or supplements...
August 11, 2017: Canadian Journal of Physiology and Pharmacology
https://www.readbyqxmd.com/read/28798045/global-mrna-polarization-regulates-translation-efficiency-in-the-intestinal-epithelium
#19
Andreas E Moor, Matan Golan, Efi E Massasa, Doron Lemze, Tomer Weizman, Rom Shenhav, Shaked Baydatch, Orel Mizrahi, Roni Winkler, Ofra Golani, Noam Stern-Ginossar, Shalev Itzkovitz
Asymmetric mRNA localization facilitates efficient translation in cells such as neurons and fibroblasts. However, the extent and significance of mRNA polarization in epithelial tissues are unclear. Here, we used single molecule transcript imaging and subcellular transcriptomics to uncover global apical-basal intracellular polarization of mRNA in the mouse intestinal epithelium. The localization of mRNAs did not generally overlap protein localization. Instead, ribosomes were more abundant on the apical sides, and apical transcripts were consequently more efficiently translated...
August 10, 2017: Science
https://www.readbyqxmd.com/read/28780391/structural-characterization-and-physicochemical-stability-profile-of-a-double-mutant-heat-labile-toxin-protein-based-adjuvant
#20
Vishal M Toprani, John M Hickey, Neha Sahni, Ronald T Toth, George A Robertson, C Russell Middaugh, Sangeeta B Joshi, David B Volkin
A novel protein adjuvant double-mutant Escherichia coli heat-labile toxin, LT (R192G/L211A) or dmLT, is in preclinical and early clinical development with various vaccine candidates. Structural characterization and formulation development of dmLT will play a key role in its successful process development, scale-up/transfer, and commercial manufacturing. This work describes extensive analytical characterization of structural integrity and physicochemical stability profile of dmLT from a lyophilized clinical formulation...
August 2, 2017: Journal of Pharmaceutical Sciences
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