An endogenous peptide that stimulates lanthanum-resistant calcium uptake in vascular tissue

1987 ◽  
Vol 65 (9) ◽  
pp. 1991-1995 ◽  
Author(s):  
William D. McCumbee ◽  
Peter Johnson ◽  
Peter J. Kasvinsky ◽  
Gary L. Wright
Keyword(s):  
High Performance ◽  
Calcium Uptake ◽  
Vascular Tissue ◽  
Paper Electrophoresis ◽  
Exchange Chromatography ◽  
Calcium Dependent ◽  
Significant Stimulation ◽  
Cation Exchange Column ◽  
Residue Concentration

A recent report has described the preparation of an extract from hemolyzed erythrocytes that has a stimulatory effect on lanthanum-resistant calcium uptake by vascular tissue in vitro and a hypertensive effect when injected into normotensive rats. The compound having a stimulatory effect on calcium uptake was further fractionated by molecular sieve and ion exchange chromatography, precipitation with CaCl2, high voltage paper electrophoresis, and high performance liquid chromatography (HPLC). HPLC yielded only a single fraction containing biological activity. This fraction was ninhydrin positive and acid labile. The amino acid composition was as follows: Asp/Asn (1.41), Ser (1.02), Glu/Gln (1.00), and Gly (2.00). Based on the assumption that the compound contains a single glutamic acid or glutamine residue, concentration–response data indicated that only nanomolar amounts of material were necessary to achieve significant stimulation. There was a marked increase in stimulatory activity of the resolubilized compound following calcium precipitation. The compound became inactive or showed a reduction in activity after being applied to a cation exchange column to remove calcium. Subsequent reprecipitation with CaCl2 and resolubilization restored the lost activity. Thus, we conclude that the compound is a small, acidic, calcium-dependent peptide that is extremely potent in stimulating lanthanum-resistant calcium uptake in vascular tissue.

Co-purification of bone resorbing activity and adenylate cyclase stimulating activity from human tumours associated with the humoral hypercalcaemia of malignancy

1987 ◽  
Vol 114 (1) ◽  
pp. 18-26 ◽  
Author(s):  
Chohei Shigeno ◽  
Itsuo Yamamoto ◽  
Shegiharu Dokoh ◽  
Megumu Hino ◽  
Jun Aoki ◽  
...  
Keyword(s):  
Bone Resorption ◽  
High Performance ◽  
Basic Protein ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Protein Factor ◽  
Human Tumours ◽  
Acid Stable ◽  
Bone Resorbing Activity

Abstract. We have partially purified a tumour factor capable of stimulating both bone resorption in vitro and cAMP accumulation in osteoblastic ROS 17/2 cells from three human tumours associated with humoral hypercalcaemia of malignancy. Purification of tumour factor by sequential acid urea extraction, gel filtration and cation-exchange chromatography, reverse-phase high performance liquid chromatography followed by analytical isoelectric focussing provided a basic protein (pI > 9.3) with a molecular weight of approximately 13 000 as a major component of the final preparation which retained both the two bioactivities. Bone resorbing activity and cAMP-increasing activity in purified factor correlated with each other. cAMP-increasing activity of the factor was heat- and acid-stable, but sensitive to alkaline ambient pH. Treatment with trypsin destroyed cAMP-increasing activity of the factor. Synthetic parathyroid hormone (PTH) antagonist, human PTH-(3– 34) completely inhibited the cAMP-increasing activity of the factor. The results suggest that this protein factor, having its effects on both osteoclastic and osteoblastic functions, may be involved in development of enhanced bone resorption in some patients with humoral hypercalcaemia of malignancy.

Determination of Taurine in Infant Formulas Using Ultrafiltration and Cation-Exchange Chromatography

1990 ◽  
Vol 73 (4) ◽  
pp. 627-631
Author(s):  
Edgar C Nicolas ◽  
Kathleen A Pfender ◽  
Michael A Aoun ◽  
Jane E Hemmer
Keyword(s):  
Human Milk ◽  
Cation Exchange ◽  
High Performance ◽  
Exchange Chromatography ◽  
Infant Formulas ◽  
Citrate Buffer ◽  
Simple Method ◽  
Cation Exchange Column ◽  
Chromatographic Resolution

Abstract A fast and simple method for determination of taurine in Infant formulas has been developed. The sample preparation uses disposable ultrafiltration cartridges to remove protein and clarify the sample. Hydrolysis Is avoided, simplifying the procedure and increasing efficiency. One mL sample Is centrlfuged In a cartridge for 45 mln. The filtrate Is diluted with pH 2.2 citrate buffer and Injected into a high performance amino acid analyzer. A cation-exchange column (sodium phase) Is used with a single buffer eluant and an Isocratic chromatographic program. Colorimetrlc detection is performed following post-column nlnhydrln reaction. Chromatographic resolution from other nlnhydrln-posltive compounds is excellent. Average recoveries for 3 levels of spike for various products were 100-102%. Precision Is 1-3% RSD, depending on product. Linearity, specificity, and ruggedness are excellent. The method Is applicable to quality control testing of milk-based, soy-based, and prehydrolyzed proteinbased Infant formulas In the ready-to-use, concentrate, and powder forms. A variety of commercially available Infant formulas from different manufacturers were analyzed and all were found to contain taurine levels comparable to human milk. Some human milk and cow's milk samples were also analyzed and results compare well with literature values

scholarly journals Brain peptides and glial growth. II. Identification of cells that secrete glia-promoting factors.

1986 ◽  
Vol 102 (3) ◽  
pp. 812-820 ◽  
Author(s):  
D Giulian ◽  
D G Young
Keyword(s):  
Nervous System ◽  
High Performance ◽  
Gel Filtration ◽  
Anion Exchange Chromatography ◽  
Brain Cell ◽  
Exchange Chromatography ◽  
Secretory Cells ◽  
Brain Peptides ◽  
The Central Nervous System

Glia-promoting factors (GPFs) are brain peptides which stimulate growth of specific macroglial populations in vitro. To identify the cellular sources of GPFs, we examined enriched brain cell cultures and cell lines derived from the nervous system for the production of growth factors. Ameboid microglia secreted astroglia-stimulating peptides, while growing neurons were the best source of the oligodendroglia-stimulating factors. These secretion products co-purified by gel filtration, anion exchange chromatography, and reverse-phase high performance liquid chromatography with GPFs isolated from goldfish and rat brain. Our findings suggest that glial growth in the central nervous system is regulated in part by a signaled release of peptides from specific secretory cells.

scholarly journals Structural Characterization and Antioxidative Activity of Low-Molecular-Weights Beta-1,3-Glucan from the Residue of ExtractedGanoderma lucidumFruiting Bodies

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Pai-Feng Kao ◽  
Shwu-Huey Wang ◽  
Wei-Ting Hung ◽  
Yu-Han Liao ◽  
Chun-Mao Lin ◽  
...  
Keyword(s):  
Cell Death ◽  
High Performance ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Water Soluble ◽  
Dependent Manner ◽  
Ros Production ◽  
Fruiting Bodies ◽  
Concentration Dependent Manner

The major cell wall constituent ofGanoderma lucidum(G. lucidum) isβ-1,3-glucan. This study examined the polysaccharide from the residues of alkaline-extracted fruiting bodies using high-performance anion-exchange chromatography (HPAEC), and it employed nuclear magnetic resonance (NMR) and mass spectrometry (MS) to confirm the structures. We have successfully isolated low-molecular-weightβ-1,3-glucan (LMG), in high yields, from the waste residue of extracted fruiting bodies ofG. lucidum. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay evaluated the capability of LMG to suppress H2O2-induced cell death in RAW264.7 cells, identifying that LMG protected cells from H2O2-induced damage. LMG treatment decreased H2O2-induced intracellular reactive oxygen species (ROS) production. LMG also influenced sphingomyelinase (SMase) activity, stimulated by cell death to induce ceramide formation, and then increase cell ROS production. Estimation of the activities of neutral and acid SMasesin vitroshowed that LMG suppressed the activities of both neutral and acid SMases in a concentration-dependent manner. These results suggest that LMG, a water-solubleβ-1,3-glucan recycled from extracted residue ofG. lucidum, possesses antioxidant capability against H2O2-induced cell death by attenuating intracellular ROS and inhibiting SMase activity.

Hypertensive factor: calcium stimulatory activity obtained from different tissues and animal species

1988 ◽  
Vol 66 (10) ◽  
pp. 1278-1281 ◽  
Author(s):  
G. L. Wright ◽  
B. S. Huang ◽  
P. J. Johnson ◽  
W. D. McCumbee
Keyword(s):  
Blood Pressure ◽  
Rat Brain ◽  
Calcium Uptake ◽  
Contractile Response ◽  
Mammalian Species ◽  
Rat Tissues ◽  
Stimulatory Activity ◽  
Significant Stimulation ◽  
Stimulation Of

It was recently shown that a peptide (hypertensive factor, HF) isolated from erythrocyte hemolysates from spontaneously hypertensive rats induced a prolonged elevation of blood pressure in normotensive rats. In addition, the peptide produced a marked stimulation of the in vitro uptake of lanthanum-resistant calcium by the aortae and enhanced the contractile response of aortic rings to constrictor agents. The present report describes findings of calcium stimulatory activity, enhancement of contractile function, or pressor activity in extracts of homogenates from several tissues of the rat and from erythrocyte hemolysates of several mammalian species. Significant stimulation of calcium uptake in aortic rings was obtained with preparations from rat brain, liver, and kidney. The activity per weight of tissue was similar for brain and kidney (approximately 2 units/g), while liver exhibited somewhat higher concentrations (4 units/g). The diffusate of cardiac tissue did not significantly alter in vitro calcium uptake by aortae. The injection of the cardiac and liver diffusates into normotensive Wistar–Kyoto rats produced slight (10 Torr) (1 Torr = 133.3 Pa) and moderate (25 Torr) elevations of blood pressure, respectively. Finally, a peptide purified from homogenates of rat brain by the protocol developed for the purification of HF from erythrocytes was shown to significantly enhance the contractile response of aortic rings to K+ and norepinephrine. Diffusates of erythrocytes from the rat, rabbit, dog, and guinea pig each caused a significant stimulation of calcium uptake and contained approximately the same level of activity (500 units/L of whole blood). Diffusates prepared from outdated human erythrocytes had no significant effect on calcium uptake, whereas those of freshly drawn samples exhibited high levels of activity. Purification of the causal compound from several rat tissues and from erythrocytes of freshly drawn human blood indicated a peptide whose amino acid composition was qualitatively similar to that of the peptide isolated from erythrocytes of rats. The results suggest that a peptide or family of similar compounds may be present in a variety of tissues of the rat and occurs in the erythrocytes of several mammalian species. These peptides influence blood pressure, but we suggest that their principal role is in the regulation of cellular calcium metabolism.

Purification of 3 monomeric monocot mannose-binding lectins and their evaluation for antipoxviral activity: potential applications in multiple viral diseases caused by enveloped viruses

2007 ◽  
Vol 85 (1) ◽  
pp. 88-95 ◽  
Author(s):  
Amandeep Kaur ◽  
Sukhdev Singh Kamboj ◽  
Jatinder Singh ◽  
Rajinder Singh ◽  
Melissa Abrahams ◽  
...  
Keyword(s):  
Cell Line ◽  
High Performance ◽  
Human Cancer ◽  
Gel Filtration ◽  
Cancer Cell Line ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Gloriosa Superba ◽  
African Green Monkey Kidney

Three monomeric monocot lectins from Zephyranthes carinata, Zephyranthes candida, and Gloriosa superba with carbohydrate specificity towards mannose derivatives and (or) oligomannose have been isolated and purified from their storage tissues. The lectins were purified by anion-exchange chromatography on DEAE–Sephacyl and by gel filtration chromatography on Biogel P-200 followed by high-performance liquid chromatography. The purified lectins, Z. carinata, Z. candida, and G. superba had molecular masses of 12, 11.5, and 12.5 kDa, respectively, as determined by gel filtration and SDS–PAGE, indicating that they are monomers. In a hapten inhibition assay, methyl-α-d-mannopyranoside inhibited agglutination of both Z. candida and Z. carinata; the latter was also inhibited by Man(α1-2)Man and Man(α1-3)Man. Gloriosa superba showed inhibition only with Man(α1-4)Man of all of the sugars and glycoproteins tested. All purified lectins agglutinated red blood cells from rabbit, whereas G. superba was also reactive towards erythrocytes from guinea pig. All of the lectins were nonglycosylated and did not require metal ions for their activity. They were labile above 60 °C and were affected by denaturing agents such as urea, thiourea, and guanidine–HCl. The lectins were virtually nonmitogenic, like other members of Amaryllidaceae and Liliaceae. Of the 3 lectins, G. superba was found to be highly toxic to the BSC-1 cell line (African green monkey kidney epithelial cells), while both of the Zephyranthes species showed significant in vitro inhibition of poxvirus replication in BSC-1 cells without any toxic effects to the cells. In addition, Z. candida also exhibited significant anticancer activity against SNB-78, a CNS human cancer cell line.

scholarly journals Potential Role of Epithelial Cell-Derived Histone H1 Proteins in Innate Antimicrobial Defense in the Human Gastrointestinal Tract

1998 ◽  
Vol 66 (7) ◽  
pp. 3255-3263 ◽  
Author(s):  
F. R. A. J. Rose ◽  
K. Bailey ◽  
J. W. Keyte ◽  
W. C. Chan ◽  
D. Greenwood ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Gastrointestinal Tract ◽  
Epithelial Cells ◽  
High Performance ◽  
Histone H1 ◽  
Exchange Chromatography ◽  
Human Gastrointestinal Tract ◽  
Proximal Small Intestine ◽  
Colonic Lumen

ABSTRACT In the human gastrointestinal tract, microorganisms are present in large numbers in the colon but are sparse in the proximal small intestine. In this study, we have shown that acid extracts of fresh human terminal ileal mucosal samples mediate antimicrobial activity. Following cation-exchange chromatography, one of the eluted fractions demonstrated antibacterial activity against bacteria normally resident in the human colonic lumen. This activity was further fractionated by reverse-phase high-performance liquid chromatography and identified as histone H1 and its fragments. We have also shown that in tissue sections, immunoreactive histone H1 is present in the cytoplasm of villus epithelial cells. In vitro culturing of detached (from the basement membrane) villus epithelial cells led to the release of antimicrobial histone H1 proteins, while the cells demonstrated ultrastructural features of programmed cell death. Our studies suggest that cytoplasmic histone H1 may provide protection against penetration by microorganisms into villus epithelial cells. Moreover, intestinal epithelial cells released into the lumen may mediate antimicrobial activity by releasing histone H1 proteins and their fragments.

Stimulation of aortic tissue calcium uptake by an extract of spontaneously hypertensive rats erythrocytes possessing hypertensive properties

1986 ◽  
Vol 64 (12) ◽  
pp. 1515-1520 ◽  
Author(s):  
G. L. Wright ◽  
M. E. Rogerson ◽  
W. D. McCumbee
Keyword(s):  
Spontaneously Hypertensive Rats ◽  
Calcium Uptake ◽  
Vascular Tissue ◽  
Aortic Tissue ◽  
Hypertensive Rats ◽  
Spontaneously Hypertensive ◽  
High K ◽  
Tissue Sensitivity ◽  
Tissue Calcium

In earlier reports we have described the isolation of a fraction from the erythrocytes of spontaneously hypertensive rats that produced hypertension when administered to normotensive rats. In addition, it was found that the fraction stimulated the uptake of "lanthanum-resistant" calcium by aortic rings excised from normotensive rats. In these studies we have found that the fraction causes a greater increase in the in vitro uptake of calcium by aortic tissue than that produced by depolarization of the tissue with high K+ or the receptor-mediated influx of calcium induced with norepinephrine. The hypertensive fraction appeared to be more effective in promoting increased calcium uptake in rabbit than in rat aortic tissue, suggesting that significant differences in tissue sensitivity to the active compound(s) may exist between species. In addition, we obtained evidence indicating that the tissue sensitivity to the action of the hypertensive fraction was greater in aortae from spontaneously hypertensive rats than from those of normotensive animals. Attempts to block the action of the hypertensive fraction with verapamil, nifedipine, and sodium nitroprusside had no significant effect on the elevation in tissue calcium. It was found, however, that the action of the hypertensive fraction was temperature dependent with reduced activity at lower temperatures. The data suggest that a compound(s) is present in the erythrocytes of rats that may have a marked effect on vascular tissue metabolism of calcium.

scholarly journals New Amylase Inhibitor Present in Corn Seeds Active In Vitro Against Amylase from Fusarium verticillioides

Plant Disease ◽  
2003 ◽  
Vol 87 (3) ◽  
pp. 233-240 ◽  
Author(s):  
Edson L. Z. Figueira ◽  
Alejandro Blanco-Labra ◽  
Antônio Carlos Gerage ◽  
Elisabete Y. S. Ono ◽  
Elizabeth Mendiola-Olaya ◽  
...  
Keyword(s):  
High Performance ◽  
Fusarium Verticillioides ◽  
Gel Filtration ◽  
Anion Exchange Chromatography ◽  
Human Saliva ◽  
Exchange Chromatography ◽  
Experimental Conditions ◽  
Amylase Inhibitor ◽  
Fusarium Sp

A screening for specific amylase inhibitor levels against amylase from Fusarium verticillioides (Fusarium moniliforme), the most relevant mycotoxigenic fungus in corn, was conducted on 37 corn hybrids. The amylase inhibitor levels in these hybrids ranged from 5.5 to 16.0 amylase inhibitor units per gram of corn (AIU/g) in the MASTER and AG5011 hybrids, respectively. The hybrid with the maximum content of inhibitor was used as the source of this new protein. The inhibitor was partially purified using fractional precipitation, gel filtration on Sephadex G-75 column, high performance liquid chromatography (HPLC) Superose HR 10/30 column, and HPLC anion exchange chromatography, obtaining a 20.7-fold purification. Electrophoresis after denaturing and heating under reductive conditions showed an apparent 23.8 kDa molecular mass and an acidic isoelectric point of 5.4, which differs from previous molecular masses reported for other inhibitors present in corn seeds (14 and 22 kDa). This inhibitor showed activity against amylases from human saliva and pancreas, from the fungi F. verticillioides and Aspergillus flavus, and from the insects Acanthoscelides obtectus, Zabrotes subfasciatus, Tribolium castaneum, and Sitotroga cerealella. The mycoflora found in the corn grain indicated Fusarium sp. as the most prevalent fungi (81.1% of the samples), with a count ranging from 1.5 × 102 to 2.4 × 106 CFU/g of corn. The presence of fumonisins was detected in 21 out of the 37 hybrids studied, ranging from 0.05 to 2.67 μg of FB per gram of corn. No correlation could be established between this amylase inhibitor level in the corn seeds and the presence of Fusarium sp. or with the fumonisin content under the experimental conditions of the test.

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