Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

1-(2-Chlorophenyl-N-methylpropyl)-3-isoquinolinecarboxamide (PK11195) is a proven enhancer of apoptotic cell death in a variety of cellular models. This effect is independent of its established cellular target, the mitochondrial benzodiazepine receptor (mBzR), since it is able to promote cell death also in mBzR knockout cells. Thus recently it was suggested that PK11195 might exert its effect by modulating the expression and function of the oncogene Bcl-2. We have previously demonstrated that Bcl-2 modulates cellular Ca2+ homeostasis as its overexpression reduces the Ca2+ concentration in the endoplasmic reticulum (ER) ([Ca2+](er)), impairing mitochondrial and cytosolic Ca2+ overload during cellular stress and therefore inhibiting the induction of the apoptotic cascade. Here, using ER, mitochondria and cytosolic targeted aequorin probes, we show that cellular treatment with PK11195 induces opposite changes in cellular Ca2+ homeostasis, increasing the [Ca2+](er) and amplifying IP(3) induced Ca2+ transients in mitochondria ([Ca2+](m)) and cytosol ([Ca2+](c)). This work provides evidence for a novel pharmacological effect of PK11195 on Ca2+ signalling which may be linked to its effect on Bcl-2 and account for its role in apoptotic cell death.
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PMID:Modulation of intracellular Ca2+ signalling in HeLa cells by the apoptotic cell death enhancer PK11195. 1892 43

In uremic patients, hyperphosphatemia is associated with cardiovascular calcification and increased cardiovascular mortality. Despite the use of phosphate binders, only half of hemodialysis (HD) patients achieve recommended serum phosphate levels. A hyperphosphoric salivary content, which correlates linearly with serum phosphate, has been reported in HD patients. We hypothesized that binding salivary phosphate during periods of fasting in addition to using phosphate binders with meals could improve the treatment of hyperphosphatemia. We assessed the phosphate-binding capacity of the natural polymer chitosan by (31)P nuclear magnetic resonance and established that 10 and 20% (wt/vol) middle viscosity chitosan solutions bind 30 and 50% of the phosphate contained in PBS, respectively. Thirteen HD patients with serum phosphate levels >6.0 mg/dl despite treatment with sevelamer hydrochloride chewed 20 mg of chitosan-loaded chewing gum twice daily for 2 wk at fast in addition to their prescribed phosphate-binding regimen. Salivary phosphate and serum phosphate significantly decreased during the first week of chewing; by the end of 2 wk, salivary phosphate decreased 55% from baseline (73.21 +/- 19.19 to 33.19 +/- 6.53; P < 0.00001), and serum phosphate decreased 31% from baseline (7.60 +/- 0.91 to 5.25 +/- 0.89 mg/dl; P < 0.00001). Salivary phosphate returned to baseline by day 15 after discontinuing the chewing gum, whereas serum phosphate levels took 30 d to return to baseline. Parathyroid hormone and serum calcium concentrations were not affected by the gum. In conclusion, adding salivary phosphate binding to traditional phosphate binders could be a useful approach for improving treatment of hyperphosphatemia in HD patients.
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PMID:Salivary phosphate-binding chewing gum reduces hyperphosphatemia in dialysis patients. 2440 15

This study examined whether collagen association by an endodontic isolate of Enterococcus faecalis conferred resistance to the bacterium against calcium hydroxide. E. faecalis A197A was grown at 46 degrees C until early stationary phase. Standardized bacterial suspensions were pretreated for 1 hour either with acid-soluble collagen or acidified phosphate-buffered saline (ac-PBS) and cultured to determine the baseline viable bacterial numbers. The bacterial suspensions were challenged with calcium hydroxide solution. Samples were removed at 6, 12, and 24 hours and cultured on tryptone soy agar plates. An adherence assay was performed to confirm that the collagen in the pretreatment medium was bound by the bacteria. Significantly more bacteria were cultivated at 12 hours in the collagen-pretreated group than the ac-PBS-pretreated group (p < 0.01). No bacteria could be cultivated at 24 hours in either group. Collagen association by E. faecalis A197A was found to increase the tolerance of the bacterium to calcium hydroxide.
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PMID:The resistance of collagen-associated, planktonic cells of Enterococcus faecalis to calcium hydroxide. 1908 23

When environmental analysis is performed, the high number of samples required and handling conditions during the transport of these samples to the laboratory are common problems. The comet assay is a useful, highly sensitive tool in biomonitoring. Some studies in the literature aim to preserve slides in lysis solution for use in the comet assay. Until now, however, no efficient methodology for preserving blood samples for this assay has been described. Because of this, the present report aimed to establish the proper conditions for samples maintenance prior to comet assay analysis. Samples were conserved in three different solutions: a high protein concentration solution (fetal bovine serum-FBS), an anticoagulant agent (a calcium chelator - ethylenediaminetetracetic acid - EDTA), and a salt buffered solution (phosphate buffered saline-PBS). Therefore, peripheral blood samples of Rhamdia quelen specimens were collected and maintained in these solutions until testing at 72h. Analyses of DNA fragmentation via the comet assay and cell viability via flow cytometry were performed at intervals of 24h. The results showed that samples maintained in FBS were preserved better; this was followed by those preserved in PBS and then last by those preserved in EDTA. In conclusion, blood samples from freshwater fish can be preserved up to 48h in fetal bovine serum at 4 degrees C in the absence of light. In this period, no DNA fragmentation occurs. We thus describe an excellent method of sample conservation for subsequent analysis in the laboratory.
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PMID:Establishment of experimental conditions for preserving samples of fish blood for analysis with both comet assay and flow cytometry. 1910 2

In a search for genes involved in regulation of uterine contractility, we cloned a novel calcium-activated chloride channel gene, named rat Clca4, from pregnant rat uterus. The gene shares approximately 83% and 70% nucleotide homology with mouse Clca6 and human CLCA4, respectively, and was expressed primarily in rat uterus. The transcripts were upregulated at Gestational Day 22 (prior to parturition), implying a functional involvement in parturition. Western blot analysis showed that rat CLCA4 protein was present in uterus, lung, and heart, but not in any other tissues examined. Confocal microscopy revealed that rat CLCA4 is localized in cell membrane and could not be removed by alkaline or PBS washing. Transient transfection of rat CLCA4-enhanced green fluorescent protein in Chinese hamster ovary cells resulted in production of characteristic Cl(-) currents that could be activated by Ca(2+) and ionomycin but inhibited by niflumic acid, a CLCA-channel blocker. The identification and characterization of rat Clca4 help decipher the contribution of Ca(2+)-activated Cl(-) conductance in myometrial contractility.
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PMID:Cloning and characterization of a calcium-activated chloride channel in rat uterus. 1914 63

A simple sensor method was developed for aflatoxin M(1) analysis to be applied directly with milk by using antibody modified screen-printed carbon working electrode with carbon counter and silver-silver chloride pseudo-reference electrode. A competitive ELISA assay format was constructed on the surface of the working electrode using 3,3,5',5'-tetramethylbenzidine dihyrochloride (TMB)/H(2)O(2) electrochemical detection scheme with horseradish peroxidase (HRP) as the enzyme label. The performance of the assay and the sensor was optimised and characterised in pure buffer conditions before applying to milk samples. Extensive interference to the electroanalytical signal was observed upon the analysis of milk. Through a series of chemical fractionations of the milk, and testing the electrochemical properties of the fractions, the interference was attributed to whey proteins with focus towards alpha-lactalbumin. A simple pre-treatment technique of incorporating 18 mM calcium chloride, in the form of Dulbucco's PBS, in a 1:1 ratio to the milk sample or standards and also to the washing buffer stabilised the whey proteins in solution and eliminate the interfering signal. The resulting immunosensor was interference free and achieved a limit of detection of 39 ng l(-1) with a linear dynamic detection range up to 1000 ng l(-1). The developed immunosensor method was compared to a commercial ELISA kit and an in-house HPLC method. The immunsensor was comparable, in term of sensitivity, but vastly superior in term of portability and cost therefore a key instrument for the detection of aflatoxin M(1) at the source of the contamination.
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PMID:Development of an electrochemical immunosensor for aflatoxin M1 in milk with focus on matrix interference. 1916 7

Encapsulation of imaging agents and drugs in calcium phosphate nanoparticles (CPNPs) has potential as a nontoxic, bioresorbable vehicle for drug delivery to cells and tumors. The objectives of this study were to develop a calcium phosphate nanoparticle encapsulation system for organic dyes and therapeutic drugs so that advanced fluoresence methods could be used to assess the efficiency of drug delivery and possible mechanisms of nanoparticle bioabsorption. Highly concentrated CPNPs encapsulating a variety of organic fluorophores were successfully synthesized. Well-dispersed CPNPs encapsulating Cy3 amidite exhibited nearly a 5-fold increase in fluorescence quantum yield when compared to the free dye in PBS. FCS diffusion data and cell staining were used to show pH-dependent dissolution of the particles and cellular uptake, respectively. Furthermore, an experimental hydrophobic cell growth inhibitor, ceramide, was successfully delivered in vitro to human vascular smooth muscle cells via encapsulation in CPNPs. These studies demonstrate that CPNPs are effective carriers of dyes and drugs for bioimaging and, potentially, for therapeutic intervention.
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PMID:Encapsulation of organic molecules in calcium phosphate nanocomposite particles for intracellular imaging and drug delivery. 1936 37

Traumatic brain injury (TBI) induces physical, cognitive, and psychosocial deficits that affect millions of patients. TBI activates numerous cellular mechanisms and molecular cascades that produce detrimental outcomes, including neuronal death and loss of function. The mitochondrion is one of the major targets of TBI, as seen by increased mitochondrial activity in activated and proliferating microglia (due to high energy requirements and/or calcium overload) as well as increased reactive oxygen species, changes in mitochondrial permeability transition, release of cytochrome c, caspase activation, reduced ATP levels, and cell death in neurons. Translocator protein (TSPO) is an 18-kDa outer mitochondrial membrane protein that interacts with the mitochondria permeability transition pore and binds with high affinity to cholesterol and various classes of drug ligands, including some benzodiazepines such as 4'-chlorodiazepam (Ro5-4864). Although TSPO levels in the brain are low, they are increased after brain injury and inflammation. This finding has led to the proposed use of TSPO expression as a marker of brain injury and repair. TSPO drug ligands have been shown to participate in the control of mitochondrial respiration and function, mitochondrial steroid and neurosteroid formation, as well as apoptosis. This review and commentary will outline our current knowledge of the benefits of targeting TSPO for TBI treatment and the mechanisms underlying the neuroprotective effects of TSPO drug ligands in neurotrauma.
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PMID:Translocator protein (18 kDa) TSPO: an emerging therapeutic target in neurotrauma. 1940 85

In leukocytes, as in many other cell types, cytoplasmic calcium ([Ca(2+)](i)) changes play a key role in a series of pathways leading to activation. Here we describe a flow cytometric method allowing the simultaneous kinetic analysis of changes in [Ca(2+)](i) in the three types of leukocytes, i.e. monocytes, granulocytes and lymphocytes. Heparinised whole blood was diluted in phosphate buffered saline with Ca(2+) and 1 mM sodium pyruvate and incubated with the Ca(2+) indicator fluo3-acetoxymethyl ester. Leukocytes were identified by labelling with the phycoerythrin-conjugated antibody against CD45, the leukocyte common antigen. Resuspension of the cells in PBS with or without Ca(2+) allowed us to detect the origin of Ca(2+) changes. During flow cytometric analysis only CD45-positive cells were counted and monocytes, granulocytes and lymphocytes were evaluated separately. Baseline fluorescence of the fluo3-Ca(2+)-complex was determined and changes in [Ca(2+)](i) after stimulation with the calcium ionophore A23187 or the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP) were recorded over a time period of 150 s. Stimulation with A23187 resulted in a rise in [Ca(2+)](i) in all three leukocyte subpopulations. This rise was sustained in the presence of extracellular Ca(2+) (Ca(2+)(ex)) but had a transient character in the absence of Ca(2+)(ex). For fMLP, [Ca(2+)](i) changes occurred only in monocytes and granulocytes and were transient irrespective of the presence or absence of Ca(2+)(ex). In conclusion, the present method is a simple, fast and easy tool to analyse in vitro [Ca(2+)](i) changes over time in leukocytes under physiologically relevant conditions, without the need for their isolation or the lysis of erythrocytes. The whole blood approach allows a continuous interaction between the different leukocyte subpopulations and other blood components and a minimum of preparative manipulations avoids artefactual activation of the cells. A distinction can be made between Ca(2+) release from the intracellular stores and the entry of Ca(2+) from outside the cell. The approach allows to evaluate the effect of various agonists on [Ca(2+)](i) changes in leukocytes, with physiological, patho-physiological or therapeutic purposes.
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PMID:Flow cytometric calcium flux assay: evaluation of cytoplasmic calcium kinetics in whole blood leukocytes. 1961 51

The purpose of this study was to evaluate the growth patterns and osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) when seeded onto new biodegradable chitosan/polyester scaffolds. Scaffolds were obtained by melt blending chitosan with poly(butylene succinate) in a proportion of 50% (wt) each and further used to produce a fiber mesh scaffold. hBMSCs were seeded on those structures and cultured for 3 weeks under osteogenic conditions. Cells were able to reduce MTS and demonstrated increasing metabolic rates over time. SEM observations showed cell colonization at the surface as well as within the scaffolds. The presence of mineralized extracellular matrix (ECM) was successfully demonstrated by peaks corresponding to calcium and phosphorus elements detected in the EDS analysis. A further confirmation was obtained when carbonate and phosphate group peaks were identified in Fourier Transformed Infrared (FTIR) spectra. Moreover, by reverse transcriptase (RT)-PCR analysis, it was observed the expression of osteogenic gene markers, namely, Runt related transcription factor 2 (Runx2), type 1 collagen, bone sialoprotein (BSP), and osteocalcin. Chitosan-PBS (Ch-PBS) biodegradable scaffolds support the proliferation and osteogenic differentiation of hBMSCs cultured at their surface in vitro, enabling future in vivo testing for the development of bone tissue engineering therapies.
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PMID:Osteogenic differentiation of human bone marrow mesenchymal stem cells seeded on melt based chitosan scaffolds for bone tissue engineering applications. 1962 27


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