Inositol phosphates are a family of water-soluble intracellular signaling molecules derived from membrane inositol phospholipids. They undergo variety complex interconversion pathways, and their levels dynamically regulated within the cytosol in response to agonists. Relatively little is known about biological function most members this family, with exception 1,4,5-trisphosphate. Specifically, functions tetrakisphosphates largely obscure. In paper, we report that D-myo-inositol...

Polyphenols, including tannins, are phytochemicals with pronounced antimicrobial properties. We studied the activity of two hydrolysable (i) gallotannin—1,2,3,4,5-penta-O-galloyl-β-D-glucose (PGG) and (ii) ellagitannin—1,2-di-O-galloyl-4,6-valoneoyl-β-D-glucose (dGVG), applied alone in combination antibiotics against Staphylococcus aureus strain 8324-4. also evaluated effect these tannins on bacterial membrane integrity fluidity their interaction proteins lipids. A correlation between...

Cystic fibrosis (CF) airway epithelia exhibit enhanced Na+ reabsorption in parallel with diminished Cl- secretion. We tested the hypothesis that actin plays a role regulation of cloned epithelial channel (ENaC) by cystic transmembrane conductance regulator (CFTR). found immunopurified bovine tracheal CFTR coreconstituted into planar lipid bilayer alpha,beta,gamma-rat ENaC (rENaC) decreased single-channel open probability (Po) rENaC presence over 60%, significantly greater effect than was...

Anions such as Cl− and HCO3 − are well known to play an important role in glucose-stimulated insulin secretion (GSIS). In this study, we demonstrate that glucose-induced efflux from β-cells is mediated by the Ca2+-activated channel anoctamin 1 (Ano1). Ano1 expression rat demonstrated reverse transcriptase–polymerase chain reaction, western blotting, immunohistochemistry. Typical currents observed whole-cell inside-out patches presence of intracellular Ca++: at μM, current outwardly...

Pulmonary fibrosis is a devastating disease without effective treatment. Rosemary appreciated since ancient times for its medicinal properties, while biomolecules originated from the plant have an antioxidant and antifibrotic effect.The effects of Rosmarinus officinalis L. (Lamiaceae) leaves extract (RO) on bleomycin-induced lung were investigated.Male Wistar rats given single dose bleomycin (BLM, 4 mg/kg, intratracheal), RO (75 intraperitoneal) was administered 3 days later continued weeks...

Abstract The objective of the study was a comparative analysis antihemolytic activity against two Staphylococcus aureus strains (8325-4 and NCTC 5655) as well α-hemolysin membrane modifying action four hydrolysable tannins with different molecular mass flexibility: 3,6-bis- O -di- -galloyl-1,2,4-tri- -galloyl- β - d -glucose (T1), 1,2,3,4,5-penta- (T2), 3- -galloyl-1,2-valoneoyl- (T3) 1,2-di- -galloyl-4,6-valoneoyl- (T4). We showed that all compounds studied manifested effects in range 5–50...

Limited information is available regarding domains within the epithelial Na⁺ channel (ENaC) which participate in amiloride binding. We previously utilized anti-amiloride antibody (BA7.1) as a surrogate receptor to delineate amino acid residues that contact amiloride, and identified putative binding domain WYRFHY (residues 278–283) extracellular of αrENaC. Mutations were generated examine role this sequence Functional analyses wild type (wt) mutant αrENaCs performed by cRNA...

The action of aldosterone to increase apical membrane permeability in responsive epithelia is thought be due activation sodium channels. Aldosterone stimulates methylation a 95-kDa protein A6 cells, and we have previously shown that the immunopurified Na⁺ channel complex increases open probability these channels planar lipid bilayers. We report here carboxylmethylation β subunit xENaC cells. <i>In vitro</i> translated subunit, but not α or γ, serves as substrate for...

Confluent monolayers of epithelial cells grown on nonporous support form fluid-filled hemicysts called domes, which reflect active ion transport across the epithelium. Clara-like H441 lung adenocarcinoma glass supports and exposed to 50 nM dexamethasone developed domes in a time-dependent fashion. Uplifting small groups occurred within 6-12 h, well formed appeared between 24 48 after 7 days, individual started merge. Cells inside compared with those outside or not dexamethasone, differed by...

Pulmonary fibrosis is characterized by over-population and excessive activation of fibroblasts myofibroblasts disrupting normal lung structure functioning. Rosemary extract rich in carnosic acid (CA) rosmarinic (RA) was reported to cure bleomycin-(BLM)-induced pulmonary fibrosis. We demonstrate that CA decreased human fibroblast (HLF) viability with IC50 value 17.13±1.06 μM, while RA had no cytotoxic effect. In the presence 50 μM RA, dose-response for shifted 11.70±1.46 indicating synergic...

The hypothesis that actin interactions account for the signature biophysical properties of cloned epithelial Na(+) channels (ENaC) (conductance, ion selectivity, and long mean open closed times) was tested using planar lipid bilayer reconstitution patch clamp techniques. We found following. 1) In bilayers, produced a more than 2-fold decrease in single channel conductance, 5-fold increase versus K(+) permselectivity, substantial times wild-type alphabetagamma-rENaC but had no effect on...

The animal welfare principles are forcing undergraduate teaching to avoid the use of animals. Therefore, many hands-on lab sessions using laboratory animals progressively replaced by computer simulations. These versatile software simulations permit observation behavior biological systems under a great variety experimental conditions. While this versatility is important, often work even when student makes wrong assumptions, situation that poses its own pedagogical problem. Hands-on learning...

Alteration in the control of bone morphogenetic protein (BMP)-regulated genes and increased expression endothelin (ET)-1 are both believed to play important roles still incompletely understood pathobiology pulmonary vascular remodeling fibrosis. Recent studies have drawn attention contribution adventitial fibroblast activation these phenomena. Because chloride channels involved physiological function fibroblasts, we hypothesized that differentially regulated by BMPs ET. We measured ion...

Pulmonary hypertension is characterized by cellular and structural changes in the vascular wall of pulmonary arteries. We hypothesized that lysophosphatidic acid (LPA), a bioactive lipid, implicated this remodeling rat model hypoxic hypertension. Exposure Wistar rats to 10% O2 for 3 weeks induced an increase mean serum levels LPA, 40.9 (log-detransformed standard deviations: 23.4-71.7) μM versus 21.6 (11.0-42.3) matched control animal group (P = 0.037). also observed perivascular LPA...

The activity of epithelial Na + selective channels is modulated by various factors, with growing evidence that membrane lipids also participate in the regulation. In present study, Triton X-100 extracts whole cells and apical membrane-enriched preparations from cultured A6 renal were floated on continuous-sucrose-density gradients. channel protein, probed immunostaining Western blots, was detected high-density fractions gradients (between 18 30% sucrose), which contain detergent-soluble...

We have recently cloned the α subunit of a bovine amiloride-sensitive Na+ channel (αbENaC). This shares extensive homology with both rat and human αENaC subunits but shows marked divergence at C terminus beginning amino acid 584 697-residue sequence. When incorporated into planar lipid bilayers, αbENaC almost exclusively exhibits main transition to 39 picosiemens (pS) very rare 13 pS step transitions one two subconductance states (26 pS). In contrast, renal homolog ENaC (αrENaC) has that is...

Hypoxia and epithelial stretch that are commonly observed in patients with acute lung injury have been shown to promote the release of serotonin (5-hydroxytryptamine, 5-HT) vitro. However, whether 5-HT contributes decrease alveolar fluid transport, which is a hallmark injury, unknown. Thus, we investigated effect on ion transport across epithelium. caused dose-dependent inhibition amiloride-sensitive current primary rat human type II cell monolayers, but did not affect Na(+)/K(+) ATPase...