scholarly journals Antioxidant, Antibacterial, and Cytoprotective Activity of Agathi Leaf Protein

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
A. S. Zarena ◽  
Shubha Gopal ◽  
R. Vineeth
Keyword(s):  
Safety Assessment ◽  
Crude Protein ◽  
Leaf Protein ◽  
Dependent Manner ◽  
Cytoprotective Activity ◽  
Sesbania Grandiflora ◽  
Rp Hplc ◽  
Maximum Inhibition ◽  
Sds Page ◽  
Dose Dependent

In the present study a protein termed agathi leaf protein (ALP) fromSesbania grandiflora Linn. (agathi) leaves was isolated after successive precipitation with 65% ammonium sulphate followed by purification on Sephadex G 75. The column chromatography of the crude protein resulted in four peaks of which Peak I (P I) showed maximum inhibition activity against hydroxyl radical. SDS-PAGE analysis of P I indicated that the molecular weight of the protein is≈29 kDa. The purity of the protein was 98.4% as determined by RP-HPLC and showed a single peak with a retention time of 19.9 min. ALP was able to reduce oxidative damage by scavenging lipid peroxidation against erythrocyte ghost (85.50 ± 6.25%), linolenic acid (87.67 ± 3.14%) at 4.33 μM, ABTS anion (88 ± 3.22%), and DNA damage (83 ± 4.20%) at 3.44 μM in a dose-dependent manner. The purified protein offered significant protection to lymphocyte (72% at 30 min) induced damage by t-BOOH. In addition, ALP showed strong antibacterial activity againstPseudomonas aeruginosa(20 ± 3.64 mm) andStaphylococcus aureus(19 ± 1.53 mm) at 200 μg/mL. The safety assessment showed that ALP does not induce cytotoxicity towards human lymphocyte at the tested concentration of 0.8 mg/mL.

Interaction between perchlorate and nifedipine on insulin secretion from mouse pancreastic islets

1993 ◽  
Vol 13 (2) ◽  
pp. 107-117 ◽  
Author(s):  
Gerd Larsson-Nyrén ◽  
Janove Sehlin
Keyword(s):  
Insulin Secretion ◽  
Insulin Release ◽  
Specific Effect ◽  
Maximum Effect ◽  
Dependent Manner ◽  
Inhibitory Effects ◽  
Maximum Inhibition ◽  
Second Phases ◽  
Dose Dependent ◽  
Higher Sensitivity

In order to elucidate the mechanisms responsible for the stimulatory effect of perchlorate (ClO4−) on insulin secretion, we have investigated the interaction between this chaotropic anion and the organic calcium antagonist nifedipine. This drug, known as a blocker of L-type calcium channels, was chosen as a tool to test the idea that ClO4− acts on insulin secretion by stimulating the gating of voltage-controlled Ca2+ channels. ClO4− amplified the stimulatory effect of D-glucose on insulin release from perfused pancreas (first and second phases) as well as from isolated islets incubated in static incubations for 60 min. This indicates that ClO4− amplifies physiologically regulated insulin secretion. Nifedipine reduced D-glucose-induced (20 mM) insulin release in a dose-dependent manner with half-maximum effect at about 0.8 μM and apparent maximum effect at 5 μM nifedipine. In the presence of 20 mM D-glucose, the inhibitory effects of 0.5, 1 or 5 μM nifedipine were only slightly, if at all, counteracted by perchlorate. When 12 mM ClO4− and 20 mM D-glucose were combined, calculation of the specific effect of ClO4− revealed that nifedipine produced almost maximum inhibition already at 0.05 μM. Thus, the perchlorate-induced amplification of D-glucose-stimulated insulin release shows higher sensitivity to nifedipine than the D-glucose-effect as such. This supports the hypothesis that perchlorate primarily affects the voltage-sensitive L-type calcium channel in the β-cell.

L(+)-Lactate Binding to a Protein in Rat Skeletal Muscle Plasma Membranes

1995 ◽  
Vol 20 (1) ◽  
pp. 112-124 ◽  
Author(s):  
Karl J. A. McCullagh ◽  
Arend Bonen
Keyword(s):  
Skeletal Muscle ◽  
Plasma Membranes ◽  
Mass Range ◽  
Dependent Manner ◽  
Rat Skeletal Muscle ◽  
Lactate Transport ◽  
Sds Page ◽  
Lactate And Pyruvate ◽  
Potential Inhibitors ◽  
Dose Dependent

Biochemical studies were conducted to determine the location of a putative lactate transport protein in rat skeletal muscle plasma membranes (PM). PM (50-100 μg protein) were incubated with [U-14C] L(+)-lactate, in the presence or absence of unlabeled monocarboxylates or potential inhibitors, after which proteins were separated by SDS-PAGE. Gel slices (2 mm) were cut and analyzed for14C. [U-14C] L(+)-lactate was bound to plasma membranes in the 30 to 40 kDa molecular mass range. Binding of [U-14C] L(+)-lactate was inhibited by N-ethylmaleimide, unlabeled L-lactate and pyruvate, and in a dose dependent manner by α-cyano-4-hydroxycinnamate (r = 0.995), but not by cytochalasin-B. The inhibition of [U-14C] L(+)-lactate binding was similar to the inhibition of lactate transport. Therefore the transport of L(+)-lactate across skeletal muscle plasma membranes involves a polypeptide of 30 to 40 kDa. Key words: transport, affinity labeling

scholarly journals Prevention of atherosclerosis by specific AT1-receptor blockade with candesartan cilexetil

2001 ◽  
Vol 2 (1_suppl) ◽  
pp. S77-S80
Author(s):  
Vasilios Papademetriou ◽  
Philippe Mammillot ◽  
Robert Redman ◽  
Aldo Notargiacomo ◽  
Puneet Narayan ◽  
...  
Keyword(s):  
At1 Receptor ◽  
Receptor Blockade ◽  
Dependent Manner ◽  
Vascular Events ◽  
Oxidised Ldl ◽  
Atherosclerotic Plaque Formation ◽  
Maximum Inhibition ◽  
Prevention Of Atherosclerosis ◽  
Dose Dependent

Several studies indicate that blockade of the renin-angiotensin-aldosterone system (RAAS) can prevent atherosclerosis and vascular events, but the precise mechanisms involved are still unclear. In this study, we investigated the effect of the AT 1-receptor blocker, candesartan, in the prevention of atherosclerosis in Watanabe heritable hyperlipidaemic (WHHL) rabbits and also the effect of AT1-receptor blockade in the uptake of oxidised LDL by macrophage cell cultures. In the first set of experiments, 12 WHHL rabbits were randomly assigned to three groups: placebo, atenolol 5 mg/kg daily or candesartan 2 mg/kg daily for six months. Compared with controls and atenolol-treated rabbits, candesartan treatment resulted in a significant 50—60% reduction of atherosclerotic plaque formation and a 66% reduction in cholesterol accumulation in the thoracic aorta. Studies in macrophage cultures indicated that candesartan prevented uptake of oxidised LDL-(oxLDL)-cholesterol by cultured macrophages. Candesartan inhibited the uptake of oxLDL in a dose-dependent manner, reaching a maximum inhibition of 70% at concentrations of 5.6 µg/ml. Further studies in other animal models and well-designed trials in humans are warranted to further explore the role of AT1-receptor blockade in the prevention of atherosclerosis.

scholarly journals Measurement of tissue transglutaminase activity in a permeabilized cell system: its regulation by Ca2+ and nucleotides

1996 ◽  
Vol 313 (3) ◽  
pp. 803-808 ◽  
Author(s):  
Peter A. SMETHURST ◽  
Martin GRIFFIN
Keyword(s):  
Tissue Transglutaminase ◽  
Cell System ◽  
Dependent Manner ◽  
Human Endothelial Cells ◽  
Permeabilized Cell ◽  
Protein Substrates ◽  
Endogenous Protein ◽  
Sds Page ◽  
Competitive Inhibitors ◽  
Dose Dependent

Electropermeabilized human endothelial cells (ECV-304) were used to study the regulation of tissue transglutaminase (tTGase) activity in the intracellular environment. An ELSA (enzyme-linked sorbent assay) plate assay was developed for intracellular tTGase activity, using the incorporation of a biotinylated primary amine, 5-{[(N-biotinoylamino)hexanoyl]amino}pentylamine (biotin-x-cadaverine; BTC), into endogenous protein substrates of tTGase. This incorporation process was inhibited by competitive inhibitors of tTGase, cystamine and monodansylcadaverine, in a dose-dependent manner. Over a 30 min period tTGase and its protein substrates did not leak out of the cell, and no incorporation of BTC occurred in unpermeabilized cells, indicating the reaction to be intracellular. In the presence of 10 nM or 10 μM Ca2+, when nucleotides ATP and GTP were added at concentrations mimicking cytosolic levels, tTGase activity was decreased virtually to zero. Only at 100 μM Ca2+, when nucleotides were low or absent was tTGase activity observed. Under these conditions a variety of proteins was labelled by the enzyme, with the major labelling found in a protein of molecular mass around 51 kDa when analysed by SDS/PAGE/Western blotting.

Human mast cells recovered by bronchoalveolar lavage: their morphology, histamine release and the effects of sodium cromoglycate

1985 ◽  
Vol 68 (4) ◽  
pp. 427-432 ◽  
Author(s):  
K. C. Flint ◽  
K. B. P. Leung ◽  
F. L. Pearce ◽  
B. N. Hudspith ◽  
J. Brostoff ◽  
...  
Keyword(s):  
Mast Cells ◽  
Bronchoalveolar Lavage ◽  
Histamine Release ◽  
Immunoglobulin E ◽  
Morphological Characteristics ◽  
Dependent Manner ◽  
Disodium Cromoglycate ◽  
Mucosal Mast Cells ◽  
Maximum Inhibition ◽  
Dose Dependent

1. Mast cells make up between 0.5 and 3% (mean 1.35%) of total cells recovered by bronchoalveolar lavage (BAL). 2. The majority of these cells have the morphological characteristics of mucosal mast cells in that they fail to stain in the alcian blue-safranin reaction after fixation in formol-saline but stain well after fixation in Carnoy's solution. Cells staining with berberine sulphate were seen in only four of the 26 lavages. 3. BAL cells released histamine in response to anti-human immunoglobulin E (IgE) in a dose-dependent manner that was optimal at a dilution of anti-IgE of 1:100. Maximum release was obtained by 2 min. 4. Histamine release was completely inhibited by a combination of 2-deoxyglucose (5 mmol/l) and antimycin A (1 μmol/l). 5. Disodium cromoglycate (DSCG) significantly inhibited this histamine release at 1 mmol/l (P<0.02), 100 μmol/l (P<0.002) and 10 μmol/l (P<0.003), with maximum inhibition of 50.1% at 10 μmol/l.

Proanthocyanidins (condensed tannin) destabilise plant protein foams in a dose dependent manner

1995 ◽  
Vol 46 (6) ◽  
pp. 1101 ◽  
Author(s):  
GJ Tanner ◽  
PJ Moate ◽  
LH Davis ◽  
RH Laby ◽  
YuGuang Li ◽  
...  
Keyword(s):  
Compressive Strength ◽  
Condensed Tannin ◽  
Red Clover ◽  
Plant Protein ◽  
Leaf Protein ◽  
Dependent Manner ◽  
Forage Legume ◽  
Chemical Structures ◽  
Dose Dependent

The compressive strength and persistence of protein foams produced from extracts of legume leaves were measured. The presence of foliar proanthocyanidin (PA, condensed tannin) in species such as Onobrychis and Lotus correlated with foams of negligible compressive strength and persistence and also with a reputation for bloat safety in the field. PA purified from forage legume leaves significantly decreased the compressive strength of protein foams produced from purified red clover leaf protein. The decrease in the compressive strength was related to the concentration of added PA in a dose-dependent manner. A ratio of PA to protein of 0.1 reduced the compressive strength by approximately half relative to the strength of the PA-free control. Purified PAS with different chemical structures had similar effects on the compressive strength of protein foams. The significance of these observations to pasture bloat and forage legume improvement is discussed.

scholarly journals Whole Peptidoglycan Extracts from theLactobacillus paracaseisubsp.paracaseiM5 Strain Exert Anticancer ActivityIn Vitro

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Shumei Wang ◽  
Xue Han ◽  
Lanwei Zhang ◽  
Yingchun Zhang ◽  
Hongbo Li ◽  
...  
Keyword(s):  
Lactobacillus Paracasei ◽  
Molar Ratio ◽  
Dependent Manner ◽  
Apoptotic Pathway ◽  
Trypan Blue Exclusion ◽  
Anticancer Effects ◽  
Transmission Electron ◽  
Sds Page ◽  
Dose Dependent ◽  
Ht 29

TheLactobacillus paracaseisubsp. paracaseiM5 strain exerted potential anticancer activity through the cell wall. In this study, whole peptidoglycan (WPG) was extracted from theLactobacillus paracaseisubsp. paracaseiM5 strain and was evaluated for anticancer effects as well as its properties. SDS-PAGE analysis confirmed the presence of WPG with dominant bands of approximately 14.4 kDa. Further analysis revealed that the amino acids present in the WPG consisted of alanine, glycine, glutamic acid, and lysine in a molar ratio of approximately 8 : 5 : 3 : 3.5. In addition, the cell viability of HT-29 cells with WPG addition was investigated with methyl thiazolyl tetrazolium (MTT) and trypan blue exclusion (TBE) assays, and cell apoptosis was analyzed with a transmission electron microscope, flow cytometry, and semiquantitative RT-PCR. These results showed that WPG exerted cytotoxic effects on colon cancer HT-29 cells in a dose-dependent manner and upregulated proapoptotic genes, while downregulating antiapoptotic genes. The gene expression study definitively revealed that WPG induced a mitochondria-mediated apoptotic pathway.

Leishmanolysin (gp63 metallopeptidase)-like activity extracellularly released byHerpetomonas samuelpessoai

Parasitology ◽  
2005 ◽  
Vol 132 (1) ◽  
pp. 37-47 ◽  
Author(s):  
C. G. R. ELIAS ◽  
F. M. PEREIRA ◽  
B. A. SILVA ◽  
C. S. ALVIANO ◽  
R. M. A. SOARES ◽  
...  
Keyword(s):  
Proteolytic Activity ◽  
Protein Profile ◽  
Immunoblot Analysis ◽  
Secretory Protein ◽  
Biochemical Properties ◽  
Maximum Activity ◽  
Dependent Manner ◽  
Sds Page ◽  
Dose Dependent ◽  
Anti Gp63

In previous studies, we showed thatHerpetomonas samuelpessoaiproduced a large amount of a surface-located metallopeptidase that presented similar biochemical properties to that of gp63 fromLeishmaniaspp., which is a well-known virulence factor expressed by these digenetic parasites. The present study aims to identify the proteolytic activity released by livingH. samuelpessoaicells. In this context, the parasites were incubated in phosphate buffer up to 4 h, and the supernatants were obtained by centrifugation and filtration steps and were then applied on SDS–PAGE to determine the secretory protein profile and on gelatin-SDS–PAGE to identify the proteolytic activity. The results demonstrated thatH. samuelpessoaisecreted at least 12 polypeptides and an extracellular peptidase of 66 kDa. This enzyme had its activity diminished by 1,10-phenanthroline, EDTA and EGTA. This metallopeptidase was active in a broad spectrum of pH, showing maximum activity at pH 6·0 at 37 °C. Casein was also cleaved by this secretory proteolytic enzyme, while bovine serum albumin and haemoglobin were not degraded under these conditions. Fluorescence microscopy and flow cytometry using anti-gp63 antibody against leishmanolysin ofL. amazonensisdemonstrated the presence of similar molecules on the cell-surface ofH. samuelpessoai. Moreover, immunoblot analysis showed the presence of a reactive polypeptide in the cellular extract and in the supernatant fluid ofH. samuelpessoai, which suggests immunological similarities between these two distinct trypanosomatids. The zinc-metallopeptidase inhibitor 1,10-phenanthroline was able to inhibit the secretion of the 66 kDa metallopeptidase in a dose-dependent manner, while the phospholipase C inhibitor (p-CMPS) did not alter the secretion pattern. Additionally, anti-cross-reacting determinant (CRD) antibody failed to recognize any secreted polypeptide fromH. samuelpessoai. Collectively, these results suggest that the gp63-like molecule was released from theH. samuelpessoaisurface by proteolysis instead of phospholipolysis, in a similar mechanism to that observed inLeishmania.

Dopamine inhibits vasopressin-dependent cAMP production in the rat cortical collecting duct

1998 ◽  
Vol 275 (1) ◽  
pp. F62-F67 ◽  
Author(s):  
Li Li ◽  
James A. Schafer ◽  
Keyword(s):  
Collecting Duct ◽  
Dependent Manner ◽  
Cortical Collecting Duct ◽  
Camp Production ◽  
Kinase C ◽  
Osmotic Water ◽  
Maximum Inhibition ◽  
Inhibitory G Protein ◽  
Opposing Effects ◽  
Dose Dependent

Dopamine inhibits Na+ and water reabsorption in the rat cortical collecting duct (CCD) in the presence of arginine vasopressin (AVP). This inhibition appears to involve the D4 dopamine receptor isoform, which inhibits cAMP production; however, the D1A receptor, which stimulates cAMP production, is also expressed in the CCD. To discriminate between these opposing effects, we measured cAMP production in intact CCD segments. The basal rate of cAMP production ranged from 6.5 to 10 fmol/mm of tubule length over a 7-min incubation period, and it was unaffected by either dopamine or the D1A-specific agonist fenoldopam. AVP increased cAMP production to the range of 85–153 fmol ⋅ mm−1 ⋅ 7 min−1. Whereas neither 0.1 nor 1.0 μM fenoldopam affected AVP-dependent cAMP production, dopamine reduced it in a dose-dependent manner, achieving a maximum inhibition of 50% at 10 μM. This effect was reversed by the D4 receptor antagonist clozapine but not by pimozide or spiperone (antagonists of D2 and D3 receptors) or by calphostin C or chelerythrine (inhibitors of protein kinase C). We conclude that dopamine inhibits transepithelial Na+ transport and osmotic water permeability in the presence of AVP by inhibition of cAMP production, which is mediated by the D4receptor isoform linked via the inhibitory G protein Gi.

scholarly journals Leishmania donovani promastigote: Effect of Adrenergic Agonists and Antagonist on Growth and Lipophosphoglycan Synthesis

2018 ◽  
Vol 14 (2) ◽  
pp. 111-115
Author(s):  
Kalipada Kar ◽  
Sujata Kar
Keyword(s):  
Leishmania Donovani ◽  
Drug Regimen ◽  
Defined Medium ◽  
Dependent Manner ◽  
Self Healing ◽  
Sds Page ◽  
Second Messenger Systems ◽  
Cutaneous Form ◽  
Dose Dependent

ABSTRACTBackground: Leishmania is the causative agent of a spectrum of diseases in human ranging in severity from self-healing cutaneous form to disfiguring mucocutaneous lesion to deadly visceral form of leishmaniasis. The treatment is still suffering from toxicity of the present drug regimen and the emergence of drug resistant leishmaniasis. Successful immunotherapy is yet to develop. It is beneficial to venture the parasite second messenger systems for the development of successful antileishmanial agents. The role of adrenergic receptor agonists and antagonist in the growth of L. donovani promastigotes and the synthesis of phosphorylated macromolecules were addressed in this report. Materials and Methods: The present work is entirely experimental. The effect of dibutyryl cAMP (db-cAMP), dibutyryl cGMP (db-cGMP), epinephrine, isoproterenol and propranolol on the growth of L. donovani promastigotes using defined medium were studied. Biosynthetically pulse labeling of promastigotes with [32P]-orthophosphate with or without experimental agents following SDS-PAGE and autoradiography was analyzed. Results: The growth of L. donovani promastigotes at 96 h culture was inhibited by 69%, 83, 52% and 95% with db-cAMP, epinephrine, isoproterenol and propranolol at 100 μM each, respectively in compared to control. Multiplication of parasite was stimulated by db-cGMP. The expression of lipophosphoglycan of the parasite was drastically affected in the following order with propranolol>epinephrine>cAMP>isoproterenol at 100 μM of each agent for 6 h exposure to the promastigotes. Conclusion: The growth of the parasite and the synthesis of lipophosphoglycan were significantly more inhibited by epinephrine or propranolol in a dose dependent manner than cAMP or isoproterenol.Keywords: cAMP; cGMP; epinephrine; Leishmania donovani; LPG; propronolol.

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