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https://www.readbyqxmd.com/read/28230977/extracellular-self-assembly-of-functional-and-tunable-protein-conjugates-from-bacillus-subtilis
#1
Charlie Gilbert, Mark Howarth, Colin R Harwood, Tom Ellis
The ability to stably and specifically conjugate recombinant proteins to one another is a powerful approach for engineering multifunctional enzymes, protein therapeutics and novel biological materials. While many of these applications have been illustrated through in vitro and in vivo intracellular protein conjugation methods, extracellular self-assembly of protein conjugates offers unique advantages: simplifying purification, reducing toxicity and burden, and enabling tunability. Exploiting the recently described SpyTag-SpyCatcher system, we describe here how enzymes and structural proteins can be genetically-encoded to covalently conjugate in culture media following programmable secretion from Bacillus subtilis...
February 23, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28230795/toxin-release-of-cyanobacterium-microcystis-aeruginosa-after-exposure-to-typical-tetracycline-antibiotic-contaminants
#2
Jing Ye, Yuping Du, Lumei Wang, Jingru Qian, Jiejing Chen, Qingwen Wu, Xiaojun Hu
The global usage of veterinary antibiotics is significant. Antibiotics can be released into aquatic environments and elicit toxic effects on non-target organisms. In this study, the growth characteristics and toxin release of the cyanobacterium Microcystis aeruginosa (M. aeruginosa) were examined to investigate the physiological effects of tetracycline antibiotics on aquatic life. Results showed that the degree of toxicities of the following target antibiotics was TC (tetracycline hydrochloride) > CTC (chlortetracycline hydrochloride) > OTC (oxytetracycline hydrochloride) in terms of growth parameters, EC10 (0...
February 21, 2017: Toxins
https://www.readbyqxmd.com/read/28228748/chromogranin-a-regulation-of-obesity-and-peripheral-insulin-sensitivity
#3
REVIEW
Gautam K Bandyopadhyay, Sushil K Mahata
Chromogranin A (CgA) is a prohormone and granulogenic factor in endocrine and neuroendocrine tissues, as well as in neurons, and has a regulated secretory pathway. The intracellular functions of CgA include the initiation and regulation of dense-core granule biogenesis and sequestration of hormones in neuroendocrine cells. This protein is co-stored and co-released with secreted hormones. The extracellular functions of CgA include the generation of bioactive peptides, such as pancreastatin (PST), vasostatin, WE14, catestatin (CST), and serpinin...
2017: Frontiers in Endocrinology
https://www.readbyqxmd.com/read/28228353/isochoric-and-isobaric-freezing-of-fish-muscle
#4
Gabriel Năstase, Chenang Lyu, Gideon Ukpai, Alexandru Șerban, Boris Rubinsky
We have recently shown that, a living organism, which succumbs to freezing to -4 °C in an isobaric thermodynamic system (constant atmospheric pressure), can survive freezing to -4 °C in an isochoric thermodynamic system (constant volume). It is known that the mechanism of cell damage in an isobaric system is the freezing caused increase in extracellular osmolality, and, the consequent cell dehydration. An explanation for the observed survival during isochoric freezing is the thermodynamic modeling supported hypothesis that, in the isochoric frozen solution the extracellular osmolality is comparable to the cell intracellular osmolality...
February 19, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28228338/extra-and-intracellular-calcium-signaling-pathway-s-differentially-regulate-histamine-induced-myometrial-contractions-during-early-and-mid-pregnancy-stages-in-buffaloes-bubalus-bubalis
#5
Abhishek Sharma, Udayraj P Nakade, Soumen Choudhury, Rajkumar Singh Yadav, Satish Kumar Garg
This study examines the differential role of calcium signaling pathway(s) in histamine-induced uterotonic action during early and mid-pregnancy stages in buffaloes. Compared to mid pregnancy, tonic contraction, amplitude and mean-integral tension were significantly increased by histamine to produce myometrial contraction during early pregnancy with small effects on phasic contraction and frequency. Although uterotonic action of histamine during both stages of pregnancy is sensitive to nifedipine (a L-type Ca(2+) channels blocker) and NNC55-0396 (T-type Ca(2+) channels blocker), the role of extracellular calcium seems to be more significant during mid-pregnancy as in this stage histamine produced only 9...
February 7, 2017: Animal Reproduction Science
https://www.readbyqxmd.com/read/28226216/overcoming-immune-dysregulation-with-immunoengineered-nanobiomaterials
#6
Evan A Scott, Nicholas B Karabin, Punn Augsornworawat
The immune system is governed by an immensely complex network of cells and both intracellular and extracellular molecular factors. It must respond to an ever-growing number of biochemical and biophysical inputs by eliciting appropriate and specific responses in order to maintain homeostasis. But as with any complex system, a plethora of false positives and false negatives can occur to generate dysregulated responses. Dysregulated immune responses are essential components of diverse inflammation-driven pathologies, including cancer, heart disease, and autoimmune disorders...
February 9, 2017: Annual Review of Biomedical Engineering
https://www.readbyqxmd.com/read/28225057/intracellular-drug-bioavailability-a-new-predictor-of-system-dependent-drug-disposition
#7
André Mateus, Andrea Treyer, Christine Wegler, Maria Karlgren, Pär Matsson, Per Artursson
Intracellular drug exposure is influenced by cell- and tissue-dependent expression of drug-transporting proteins and metabolizing enzymes. Here, we introduce the concept of intracellular bioavailability (Fic) as the fraction of extracellular drug available to bind intracellular targets, and we assess how Fic is affected by cellular drug disposition processes. We first investigated the impact of two essential drug transporters separately, one influx transporter (OATP1B1; SLCO1B1) and one efflux transporter (P-gp; ABCB1), in cells overexpressing these proteins...
February 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28224799/preparation-and-practical-applications-of-2-7-dichlorodihydrofluorescein-in-redox-assays
#8
Megan J Reiniers, Rowan F van Golen, Sylvestre Bonnet, Mans Broekgaarden, Thomas M van Gulik, Maarten R Egmond, Michal Heger
Oxidative stress, a state in which intra- or extracellular oxidant production outweighs the antioxidative capacity, lies at the basis of many diseases. DCFH2-DA (2',7'-dichlorodihydorfluorescein diacetate) is the most widely used fluorogenic probe for the detection of general oxidative stress. However, the use of DCFH2-DA, as many other fluorogenic redox probes, is mainly confined to the detection of intracellular oxidative stress in vitro. To expand the applicability of the probe, an alkaline hydrolysis and solvent extraction procedure was developed to generate high-purity DCFH2 (2',7'-dichlorodihydorfluorescein) from DCFH2-DA using basic laboratory equipment...
February 22, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28224564/increased-glucocorticoid-receptor-expression-in-sepsis-is-related-to-heat-shock-proteins-cytokines-and-cortisol-and-is-associated-with-increased-mortality
#9
Konstantinos Vardas, Stavroula Ilia, Amalia Sertedaki, Evangelia Charmandari, Efrossini Briassouli, Dimitris Goukos, Kleovoulos Apostolou, Katerina Psarra, Efthimia Botoula, Stylianos Tsagarakis, Eleni Magira, Christina Routsi, Constantine A Stratakis, Serafim Nanas, George Briassoulis
BACKGROUND: The purposes of this study are to examine if the human glucocorticoid receptor (hGR) isoform-α mRNA and hGR protein expressions are deficient in the acute phase of sepsis (S) compared to systemic inflammatory response syndrome (SIRS) and healthy subjects (H) and to evaluate if the hGRα and hGR alterations are associated with cortisol changes and if they are related to (1) extracellular and intracellular heat shock proteins (HSP) 72 and 90α; (2) ACTH, prolactin, and interleukins (ILs); and (3) outcome...
December 2017: Intensive Care Medicine Experimental
https://www.readbyqxmd.com/read/28223427/calcium-dependent-enhancement-by-extracellular-acidity-of-the-cytotoxicity-of-mitochondrial-inhibitors-against-melanoma
#10
Fumihito Noguchi, Shigeki Inui, Clare Fedele, Mark Shackleton, Satoshi Itami
Extracellular acidity is a hallmark of cancers and is independent of hypoxia. Since acidity potentiates malignant phenotypes, therapeutic strategies that enhance the targeting of oncogenic mechanisms in an acidic microenvironment should be effective. We report here that drugs which abrogate mitochondrial respiration show enhanced cytotoxicity against melanoma cells in a normoxic but acidic extracellular pH, independent from P53 mutations, BRAF (V600E) mutations and/or resistance against BRAF inhibitors. Conversely, the cytotoxicity against melanoma cells of mitochondrial inhibitors is impaired by a neutral or alkaline extracellular pH and in vivo systemic alkalinization with NaHCO3 enhanced subcutaneous tumor growth and lung metastasis of B16F10 cells in mice treated with the mitochondrial inhibitor phenformin...
February 21, 2017: Molecular Cancer Therapeutics
https://www.readbyqxmd.com/read/28223252/the-origin-and-evolution-of-basigin-bsg-gene-a-comparative-genomic-and-phylogenetic-analysis
#11
Xinyan Zhu, Shenglan Wang, Mingjie Shao, Jie Yan, Fei Liu
Basigin (BSG), also known as extracellular matrix metalloproteinase inducer (EMMPRIN) or cluster of differentiation 147 (CD147), plays various fundamental roles in the intercellular recognition involved in immunologic phenomena, differentiation, and development. In this study, we aimed to compare the similarities and differences of BSG among organisms and explore possible evolutionary relationships based on the comparison result. We used the extensive BLAST tool to search the metazoan genomes, N-glycosylation sites, the transmembrane region and other functional sites...
February 18, 2017: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/28223223/berberine-improves-cognitive-impairment-by-promoting-autophagic-clearance-and-inhibiting-production-of-%C3%AE-amyloid-in-app-tau-ps1-mouse-model-of-alzheimer-s-disease
#12
Min Huang, Xin Jiang, Yubin Liang, Qiong Liu, Siyan Chen, Yi Guo
This study investigates the neuroprotective properties of berberine (a natural isoquinoline alkaloid isolated from the Rhizoma coptidis) and finds that berberine could promote β-amyloid (Aβ) clearance and inhibit Aβ production in the triple-transgenic mouse model of Alzheimer's disease (3×Tg-AD). During the study, berberine was first administrated to treat 3×Tg-AD mice and primary neurons. Morris water maze assay, western blotting, enzyme-linked immunosorbent assay (ELISA), immunofluorescence staining and histological analysis, transmission electron microscopic analysis were then used to evaluate the effects of the berberine administration...
February 20, 2017: Experimental Gerontology
https://www.readbyqxmd.com/read/28223156/electrophysiological-properties-of-computational-human-ventricular-cell-action-potential-models-under-acute-ischemic-conditions
#13
REVIEW
Sara Dutta, Ana Mincholé, T Alexander Quinn, Blanca Rodriguez
Acute myocardial ischemia is one of the main causes of sudden cardiac death. The mechanisms have been investigated primarily in experimental and computational studies using different animal species, but human studies remain scarce. In this study, we assess the ability of four human ventricular action potential models (Tentusscher et al., 2006; Grandi et al., 2010; Carro et al., 2011; O'Hara et al., 2011) to simulate key electrophysiological consequences of acute myocardial ischemia in single cell and tissue simulations...
February 18, 2017: Progress in Biophysics and Molecular Biology
https://www.readbyqxmd.com/read/28222887/rare-er-protein-misfolding-mistrafficking-disorders-therapeutic-developments
#14
REVIEW
Ramanath Narayana Hegde, Advait Subramanian, Prathyush Pothukuchi, Seetharaman Parashuraman, Alberto Luini
The presence of a functional protein at the appropriate location in the cell is the result of the processes of transcription, translation, folding and trafficking to the correct destination. There are numerous diseases that are caused by protein misfolding, mainly due to mutations in the respective gene. The consequences of this misfolding may be that proteins effectively lose their function, either by being removed by the cellular quality control machinery or by accumulating at the incorrect intracellular or extracellular location...
February 9, 2017: Tissue & Cell
https://www.readbyqxmd.com/read/28222432/extracellular-hmgb1-modulates-glutamate-metabolism-associated-with-kainic-acid-induced-epilepsy-like-hyperactivity-in-primary-rat-neural-cells
#15
Yuji Kaneko, Colleen Pappas, Teresita Malapira, Fernando Ĺ Vale, Naoki Tajiri, Cesar V Borlongan
BACKGROUND/AIMS: Neuroinflammatory processes have been implicated in the pathophysiology of seizure/epilepsy. High mobility group box 1 (HMGB1), a non-histone DNA binding protein, behaves like an inflammatory cytokine in response to epileptogenic insults. Kainic acid (KA) is an excitotoxic reagent commonly used to induce epilepsy in rodents. However, the molecular mechanism by which KA-induced HMGB1 affords the initiation of epilepsy, especially the role of extracellular HMGB1 in neurotransmitter expression, remains to be elucidated...
February 20, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28220076/actions-and-mechanisms-of-polyunsaturated-fatty-acids-on-voltage-gated-ion-channels
#16
REVIEW
Fredrik Elinder, Sara I Liin
Polyunsaturated fatty acids (PUFAs) act on most ion channels, thereby having significant physiological and pharmacological effects. In this review we summarize data from numerous PUFAs on voltage-gated ion channels containing one or several voltage-sensor domains, such as voltage-gated sodium (NaV), potassium (KV), calcium (CaV), and proton (HV) channels, as well as calcium-activated potassium (KCa), and transient receptor potential (TRP) channels. Some effects of fatty acids appear to be channel specific, whereas others seem to be more general...
2017: Frontiers in Physiology
https://www.readbyqxmd.com/read/28219986/functional-analysis-of-the-transmembrane-and-cytoplasmic-domains-of-pcdh15a-in-zebrafish-hair-cells
#17
Reo Maeda, Itallia V Pacentine, Timothy Erickson, Teresa Nicolson
Protocadherin 15 (PCDH15) is required for mechanotransduction in sensory hair cells as a component of the tip link. Isoforms of PCDH15 differ in their cytoplasmic domains (CD1, CD2, and CD3), but share the extracellular and transmembrane (TMD) domains, as well as an intracellular domain known as the common region (CR). In heterologous expression systems, both the TMD and CR of PCDH15 have been shown to interact with members of the mechanotransduction complex. The in vivo significance of these protein-protein interaction domains of PCDH15 in hair cells has not been determined...
February 20, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28219946/distinct-intracellular-ca-2-dynamics-regulate-apical-constriction-and-differentially-contribute-to-neural-tube-closure
#18
Makoto Suzuki, Masanao Sato, Hiroshi Koyama, Yusuke Hara, Kentaro Hayashi, Naoko Yasue, Hiromi Imamura, Toshihiko Fujimori, Takeharu Nagai, Robert E Campbell, Naoto Ueno
Early in the development of the central nervous system, progenitor cells undergo a shape change, called apical constriction, that triggers the neural plate to form a tubular structure. How apical constriction in the neural plate is controlled, and contributes to tissue morphogenesis, are not fully understood. In this study, we show that intracellular calcium ions (Ca(2+)) are required for Xenopus neural tube formation, and that there are two types of Ca(2+)-concentration changes, a single-cell and a multicellular wave-like fluctuation, in the developing neural plate...
February 20, 2017: Development
https://www.readbyqxmd.com/read/28219892/an-infectious-disease-associated-il12b-polymorphism-regulates-il-12-23-p40-transcription-involving-poly-adp-ribose-polymerase-1
#19
Quanju Zhao, Qinglin Du, Fang Wei, Jianping Xie, Xiaojing Ma
IL-12 and IL-23 are important host defense factors produced by APCs against certain intracellular and extracellular pathogens. Their dysregulation has also been implicated in several autoimmune diseases. The nucleotide polymorphism in the promoter region of Il12b (rs41292470 consisting of the long or short allele) encoding the shared subunit of IL-12 and IL-23, p40, has been reported to associate with susceptibility to infectious diseases and autoimmune disorders. How these genetic variants impact Il12b expression at the molecular level was unclear...
February 20, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28219080/survival-of-tissue-resident-memory-t-cells-requires-exogenous-lipid-uptake-and-metabolism
#20
Youdong Pan, Tian Tian, Chang Ook Park, Serena Y Lofftus, Shenglin Mei, Xing Liu, Chi Luo, John T O'Malley, Ahmed Gehad, Jessica E Teague, Sherrie J Divito, Robert Fuhlbrigge, Pere Puigserver, James G Krueger, Gökhan S Hotamisligil, Rachael A Clark, Thomas S Kupper
Tissue-resident memory T (TRM) cells persist indefinitely in epithelial barrier tissues and protect the host against pathogens. However, the biological pathways that enable the long-term survival of TRM cells are obscure. Here we show that mouse CD8(+) TRM cells generated by viral infection of the skin differentially express high levels of several molecules that mediate lipid uptake and intracellular transport, including fatty-acid-binding proteins 4 and 5 (FABP4 and FABP5). We further show that T-cell-specific deficiency of Fabp4 and Fabp5 (Fabp4/Fabp5) impairs exogenous free fatty acid (FFA) uptake by CD8(+) TRM cells and greatly reduces their long-term survival in vivo, while having no effect on the survival of central memory T (TCM) cells in lymph nodes...
February 20, 2017: Nature
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