Sodium affinity of brain Na(+)-K(+)-ATPase is dependent on isozyme and environment of the pump

1990 ◽  
Vol 258 (5) ◽  
pp. C803-C811 ◽  
Author(s):  
J. L. Brodsky ◽  
G. Guidotti
Keyword(s):  
Plasma Membranes ◽  
Fibroblast Cell ◽  
Mouse Fibroblast ◽  
Synaptic Plasma Membranes ◽  
Apparent Dissociation Constant ◽  
Low Sodium ◽  
Physiological Relevance ◽  
Mouse Fibroblast Cell Line

The sodium affinities for the two forms of the Na(+)-K(+)-ATPase in brain were characterized. To mimic physiological conditions, synaptosomes, which are pinched off presynaptic nerve termini, were used. Examination of the pump in vitro was performed by preparing synaptic plasma membranes (SPMs). It was first shown that synaptosomes contain the two forms of the Na(+)-K(+)-ATPase, alpha 1 and alpha 2, and that these forms have markedly different affinities for the inhibitory cardiac glycoside ouabain. The apparent dissociation constant (K0.5) of alpha 1 for sodium changed from 12 to 9 mM when going from synaptosomes to membranes. For alpha 2, however, a shift from 36 to 12.5 mM was evident. The conclusion is that in vivo alpha 2 exists as a low sodium affinity species but can be altered to a high-affinity form simply by vesicle disruption. By comparison, the Na(+)-K(+)-ATPase from the mouse fibroblast cell line, 3T3-F442A cells, expressed only the alpha 1-isozyme, as shown by immunoblotting and by measurement of its ouabain and sodium affinities. The physiological relevance of these observations is also presented.

scholarly journals Development of Bionanocomposites Based on PLA, Collagen and AgNPs and Characterization of Their Stability and In Vitro Biocompatibility

2020 ◽  
Vol 10 (7) ◽  
pp. 2265
Author(s):  
Maria Râpă ◽  
Laura Mihaela Stefan ◽  
Traian Zaharescu ◽  
Ana-Maria Seciu ◽  
Anca Andreea Țurcanu ◽  
...  
Keyword(s):  
Normal Growth ◽  
Fibroblast Cell ◽  
Stability Study ◽  
Mouse Fibroblast ◽  
Poly Lactic Acid ◽  
In Vitro Biocompatibility ◽  
Diphenyltetrazolium Bromide ◽  
Mouse Fibroblast Cell Line ◽  
The Stability

Bionanocomposites including poly(lactic acid) (PLA), collagen, and silver nanoparticles (AgNPs) were prepared as biocompatible and stable films. Thermal properties of the PLA-based bionanocomposites indicated an increase in the crystallinity of PLA plasticized due to a small quantity of AgNPs. The results on the stability study indicate the promising contribution of the AgNPs on the durability of PLA-based bionanocomposites. In vitro biocompatibility conducted on the mouse fibroblast cell line NCTC, clone 929, using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed high values of cell viability (>80%) after cell cultivation in the presence of bionanocomposite formulations for 48 h, while the percentages of lactate dehydrogenase (LDH) released in the culture medium were reduced (<15%), indicating no damages of the cell membranes. In addition, cell cycle analysis assessed by flow cytometry indicated that all tested bionanocomposites did not affect cell proliferation and maintained the normal growth rate of cells. The obtained results recommend the potential use of PLA-based bionanocomposites for biomedical coatings.

scholarly journals Omega-3 Fatty Acid Based Nanolipid Formulation of Atorvastatin for Treating Hyperlipidemia

2019 ◽  
Vol 9 (2) ◽  
pp. 271-280 ◽  
Author(s):  
Revathy Sreedhar ◽  
Vrinda Sasi Kumar ◽  
Anil Kumar Bhaskaran Pillai ◽  
Sabitha Mangalathillam
Keyword(s):  
Fatty Acid ◽  
Entrapment Efficiency ◽  
Tween 80 ◽  
Fibroblast Cell ◽  
Mouse Fibroblast ◽  
In Vivo Studies ◽  
Omega 3 ◽  
Omega 3 Fatty Acid

Purpose: In the current study, attempts have been made to formulate an omega-3 fatty acid based nanostructured lipid carriers of atorvastatin (AT), for treating hyperlipidemia; and to evaluate their antihyperlipidemic activity using in vitro and in vivo studies. Methods: Omega-3 fatty acid based AT-loaded nanolipid carriers (NLC) were formulated by the melt emulsification ultrasonication technology. The prepared NLC consist of stearic acid (as solid lipid), omega-3 fatty acid (as liquid lipid), Tween 80, poloxamer 188 (surfactants) and soya-lecithin (co-surfactant). Results: AT loaded NLCs have a particle size of 74.76 ± 4.266 nm, a zeta potential value of -36.03 ± 1.504 mV and a high drug entrapment efficiency (EE) of 86.70 % ± 0.155. The release of AT from NLCs exhibited a sustained behaviour, which made it an ideal vehicle for drug delivery. MTT assay results indicated that NLCs are compatible with L929 (mouse fibroblast) cell lines. Anti-hyperlipidemic study showed a significant reduction in LDL and TG levels in serum with the orally administered Omega-3 fatty acid based AT loaded NLCs when compared to marketed formulation. Conclusion: The results demonstrated that the omega-3 fatty acid based NLC has the potential to be a promising nanomedicine for the treatment of hyperlipidemia.

In vitro cytotoxicity evaluation of plastic biomedical devices

2001 ◽  
Vol 20 (8) ◽  
pp. 412-417 ◽  
Author(s):  
A B Pant ◽  
A K Agarwal ◽  
V P Sharma ◽  
P K Seth
Keyword(s):  
Growth Inhibition ◽  
Fibroblast Cell ◽  
Essential Amino Acids ◽  
In Vitro Cytotoxicity ◽  
Mouse Fibroblast ◽  
Biomedical Devices ◽  
Minimum Essential Medium ◽  
Growth Inhibition Assay ◽  
Mouse Fibroblast Cell Line

Cytotoxic potential of four plastic biomedical devices (intravenous transfusion sets, IV sets; dextrose normal saline bottles, DNS bottles; Ringer lactate bottles, RL bottles; and Ryle's tubes) including 17different brands was evaluated by investigating growth inhibition, percent survival, mitotic index and colony-forming ability (cfa) in L929, an adherent type mouse fibroblast cell line. Experimental sets were exposed with leachates of biomedical products in serumfree minimum essential medium (MEM) for 1 h at 378Cina CO2 incubator. After 1 h, medium was replaced with serumrich MEM containing essential amino acids and reincubated up to 96 h. Cells in serum-free MEM only were processed under identical conditions and served as the control. The leachates from all types of biomedical devices evaluated exhibitedreductioninthegrowthandsurvivalofthecellline in the first 12 h postexposure followed by their gradual recoveryupto96h.Asignificantreductionincellgrowthwas apparent in the six brands of IV sets from 24 h onwards up to 36 h (59% growth inhibition). Though the cfa was also reduced in all the brands tested, the magnitude of reduction was less compared to growth inhibition. The results indicate that leachates of IV sets were more toxic compared to other biomedical devices screened, and growth inhibition assay was found to be more sensitive and suitable for cytotoxicity evaluation of biomedical devices.

Survival and Function of Fibroblasts Stored as Stock Cultures at a Non-freezing Temperature

1993 ◽  
Vol 21 (2) ◽  
pp. 206-209
Author(s):  
Anders H. G. Andrén ◽  
Anders P. Wieslander
Keyword(s):  
Cell Growth ◽  
Cell Line ◽  
Cell Lines ◽  
Fibroblast Cell ◽  
Freezing Temperature ◽  
Mouse Fibroblast ◽  
Toxic Response ◽  
Normal Manner ◽  
And Function

Cytotoxicity, measured as inhibition of cell growth of cultured cell lines, is a widely used method for testing the safety of biomaterials and chemicals. One major technical disadvantage with this method is the continuous routine maintenance of the cell lines. We decided to investigate the possibility of storing stock cultures of fibroblasts (L-929) in an ordinary refrigerator as a means of reducing the routine workload. Stock cultures of the mouse fibroblast cell line L-929 were prepared in plastic vials with Eagle's minimum essential medium. The vials were stored in a refrigerator at 4–10°C for periods of 7–31 days. The condition of the cells after storage was determined as cell viability, cell growth and the toxic response to acrylamide, measured as cell growth inhibition. We found that the L-929 cell line can be stored for 2–3, weeks with a viabilty > 90% and a cell growth of about 95%, compared to L-929 cells grown and subcultured in the normal manner. The results also show that the toxic response to acrylamide, using refrigerator stored L-929 cells, corresponds to that of control L-929 cells. We concluded that it is possible to store L-929 cells in a refrigerator for periods of up to 3 weeks and still use the cells for in vitro cytotoxic assays.

scholarly journals A Proposed Fabrication Method of Novel PCL-GEL-HAp Nanocomposite Scaffolds for Bone Tissue Engineering Applications

2010 ◽  
Vol 19 (4) ◽  
pp. 096369351001900 ◽  
Author(s):  
A. Hamlekhan ◽  
M. Mozafari ◽  
N. Nezafati ◽  
M. Azami ◽  
H. Hadipour
Keyword(s):  
Tissue Engineering ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Mechanical Test ◽  
Fibroblast Cell ◽  
Mouse Fibroblast ◽  
Fabrication Method ◽  
Engineering Applications ◽  
Poly Caprolactone

In this study, poly(∊-caprolactone) (PCL), gelatin (GEL) and nanocrystalline hydroxyapatite (HAp) was applied to fabricate novel PCL-GEL-HAp nanaocomposite scaffolds through a new fabrication method. With the aim of finding the best fabrication method, after testing different methods and solvents, the best method and solvents were found, and the nanocomposites were prepared through layer solvent casting combined with freeze-drying. Acetone and distillated water were used as the PCL and GEL solvents, respectively. The mechanical test showed that the increasing of the PCL weight through the scaffolds caused the improvement of the final nanocomposite mechanical behavior due to the increasing of the ultimate stress, stiffness and elastic modulus (8 MPa for 0% wt PCL to 23.5 MPa for 50% wt PCL). The biomineralization investigation of the scaffolds revealed the formation of bone-like apatite layers after immersion in simulated body fluid (SBF). In addition, the in vitro cytotoxity of the scaffolds using L929 mouse fibroblast cell line (ATCC) indicated no sign of toxicity. These results indicated that the fabricated scaffold possesses the prerequisites for bone tissue engineering applications.

scholarly journals Cofactor bypass variants reveal a conformational control mechanism governing cell wall polymerase activity

2016 ◽  
Vol 113 (17) ◽  
pp. 4788-4793 ◽  
Author(s):  
Monica Markovski ◽  
Jessica L. Bohrhunter ◽  
Tania J. Lupoli ◽  
Tsuyoshi Uehara ◽  
Suzanne Walker ◽  
...  
Keyword(s):  
Polymerase Activity ◽  
Activation Mechanism ◽  
Phenotypic Analysis ◽  
Outer Membranes ◽  
Division Site ◽  
Physiological Relevance ◽  
Membrane Lipoproteins ◽  
Shed Light

To fortify their cytoplasmic membrane and protect it from osmotic rupture, most bacteria surround themselves with a peptidoglycan (PG) exoskeleton synthesized by the penicillin-binding proteins (PBPs). As their name implies, these proteins are the targets of penicillin and related antibiotics. We and others have shown that the PG synthases PBP1b and PBP1a ofEscherichia colirequire the outer membrane lipoproteins LpoA and LpoB, respectively, for their in vivo function. Although it has been demonstrated that LpoB activates the PG polymerization activity of PBP1b in vitro, the mechanism of activation and its physiological relevance have remained unclear. We therefore selected for variants of PBP1b (PBP1b*) that bypass the LpoB requirement for in vivo function, reasoning that they would shed light on LpoB function and its activation mechanism. Several of these PBP1b variants were isolated and displayed elevated polymerization activity in vitro, indicating that the activation of glycan polymer growth is indeed one of the relevant functions of LpoB in vivo. Moreover, the location of amino acid substitutions causing the bypass phenotype on the PBP1b structure support a model in which polymerization activation proceeds via the induction of a conformational change in PBP1b initiated by LpoB binding to its UB2H domain, followed by its transmission to the glycosyl transferase active site. Finally, phenotypic analysis of strains carrying a PBP1b* variant revealed that the PBP1b–LpoB complex is most likely not providing an important physical link between the inner and outer membranes at the division site, as has been previously proposed.

scholarly journals Cortistatin Is Not a Somatostatin Analogue but Stimulates Prolactin Release and Inhibits GH and ACTH in a Gender-Dependent Fashion: Potential Role of Ghrelin

Endocrinology ◽  
2011 ◽  
Vol 152 (12) ◽  
pp. 4800-4812 ◽  
Author(s):  
José Córdoba-Chacón ◽  
Manuel D. Gahete ◽  
Ana I. Pozo-Salas ◽  
Antonio J. Martínez-Fuentes ◽  
Luis de Lecea ◽  
...  
Keyword(s):  
Metabolic Phenotype ◽  
Somatostatin Analogue ◽  
Growth Hormone Secretagogue Receptor ◽  
Growth Hormone Secretagogue ◽  
Ancestral Gene ◽  
Ghrelin Receptor ◽  
Physiological Relevance

Cortistatin (CST) and somatostatin (SST) evolve from a common ancestral gene and share remarkable structural, pharmacological, and functional homologies. Although CST has been considered as a natural SST-analogue acting through their shared receptors (SST receptors 1–5), emerging evidence indicates that these peptides might in fact exert unique roles via selective receptors [e.g. CST, not SST, binds ghrelin receptor growth hormone secretagogue receptor type 1a (GHS-R1a)]. To determine whether the role of endogenous CST is different from SST, we characterized the endocrine-metabolic phenotype of male/female CST null mice (cort−/−) at hypothalamic-pituitary-systemic (pancreas-stomach-adrenal-liver) levels. Also, CST effects on hormone expression/secretion were evaluated in primary pituitary cell cultures from male/female mice and female primates (baboons). Specifically, CST exerted an unexpected stimulatory role on prolactin (PRL) secretion, because both male/female cort−/− mice had reduced PRL levels, and CST treatment (in vivo and in vitro) increased PRL secretion, which could be blocked by a GHS-R1a antagonist in vitro and likely relates to the decreased success of female cort−/− in first-litter pup care at weaning. In contrast, CST inhibited GH and adrenocorticotropin-hormone axes in a gender-dependent fashion. In addition, a rise in acylated ghrelin levels was observed in female cort−/− mice, which were associated with an increase in stomach ghrelin/ghrelin O-acyl transferase expression. Finally, CST deficit uncovered a gender-dependent role of this peptide in the regulation of glucose-insulin homeostasis, because male, but not female, cort−/− mice developed insulin resistance. The fact that these actions are not mimicked by SST and are strongly gender dependent offers new grounds to investigate the hitherto underestimated physiological relevance of CST in the regulation of physiological/metabolic processes.

Rosette formation by a mouse fibroblast cell line

Nature ◽  
1974 ◽  
Vol 248 (5448) ◽  
pp. 514-515 ◽  
Author(s):  
E. V. ELLIOTT ◽  
R. S. KERBEL ◽  
B. J. PHILLIPS
Keyword(s):  
Cell Line ◽  
Fibroblast Cell ◽  
Mouse Fibroblast ◽  
Rosette Formation ◽  
Fibroblast Cell Line ◽  
Mouse Fibroblast Cell ◽  
Mouse Fibroblast Cell Line
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