Venous blood was collected from clinically healthy (n = 5) and B. canis (n = 5) infected dogs. Parasitised red blood cells were not removed from blood samples prior to preparation. The erythrocytes were washed, lysed, the haem ring removed from the haemoglobin molecule and the globin chains denatured by ...

A phospholipid containing formulation with tetracaine (CAS 94-24-6) for topical anaesthesia was developed. In vitro drug liberation as well as in vivo efficacy and duration of the anaesthetic effect were investigated. The in vitro liberation constant was 0-.626 +/- 0.005 mg/(cm2.square root of h for a liposomal gel formulation containing ...

An acidic and a basic phospholipase A (PLA) and a toxin have been purified from the venom of W. aegyptia using a TSK G 3000 SW gel filtration column and a Mono Q ion-exchange column. Both phospholipases and the toxin were found to be in a homogeneous state on SDS-PAGE ...

We describe a group of basic isoforms of Par o 1 (cumulatively referred to as Par o 1b), purified by anion-exchange chromatography. The allergenic activity of Par o 1b was compared with that of the acidic isoform (Par o 1a) by RAST inhibition. Par o 1b showed a cathodic mobility ...

Azocasein, diffused into gels after polyacrylamide gel electrophoresis, provides a useful substrate for simple and rapid protease detection. After electrophoresis, the gels are incubated in an azocasein solution, immersed in trichloroacetic acid and then treated with NaOH. Proteolytic zones appear as uncolored bands on an orange background. The gels can ...

A lectin isolated from Saraca indica seed integument was purified by affinity chromatography on porcine thyroglobulin Sepharose followed by Sephadex G-50 and shown to be homogeneous by PAGE. It showed a single band on SDS-PAGE in the absence and presence of 2-mercaptoethanol corresponding to a M(r) of congruent to 12,000, ...

Addition of osmoprotective compounds has a positive effect on growth and monoclonal antibody production in hyperosmotic hybridoma cell cultures. In order to better understand the processes involved in the osmoprotective response, uptake of the osmoprotective compounds glycine betaine, proline, sarcosine and glycine in mouse hybridoma cell line 6H11 during exposure ...

Rat hepatic asialoglycoprotein receptors (ASGP-Rs) are hetero-oligomers composed of three homologous glycoprotein subunits, designated rat hepatic lectins (RHL) 1, 2, and 3. ASGP-Rs mediate the endocytosis and degradation of circulating glycoconjugates containing terminal N-acetylgalactosamine or galactose, including desialylated plasma glycoproteins. We have shown in permeable rat hepatocytes that the ligand ...

Characterization of normal airway mucus is required to elucidate mechanisms protecting the airways and to understand changes associated with disease and environmental insult. Toward this goal, we collected bronchial washes (10 ml saline) from healthy human subjects to 1) evaluate the yield of high-density material (delta > or = 1.35 ...

An esterase activity obtained from a strain of Bacillus stearothermophilus was purified 5,133-fold to electrophoretic homogeneity with 26% recovery. The purified esterase had a specific activity of 2,032 mumol min-1 mg-1 based on the hydrolysis of p-nitrophenyl caproate at pH 7.0 and 30 degrees C. The apparent molecular mass was ...

A unique family of proline-rich proteins (PRPs) is induced in rats following prolonged isoproterenol treatment. PRPs can be divided into glycosylated (GPRP), basic (BPRP) and acidic (APRP) proline-rich proteins based on their physicochemical characteristics. Inducible rat parotid PRPs were isolated from aqueous extracts of parotid glands of isoproterenol-treated animals by ...

1-Aminocyclopropane-1-carboxylic acid (ACC) can be oxidized to ethylene or diverted to the conjugate 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC) by an ACC N-malonyltransferase. We developed a facile assay for the ACC N-malonyltransferase that resolved [14C]MACC from [14C]ACC by thin-layer chromatography and detected and quantified them using a radioisotope-imaging system. Using this assay, we ...

Reduced alkylated glutenin subunits from wheat flour were fractionated by preparative electrophoresis at acid pH. The high molecular weight glutenin subunits (HMW-GS) and some of the low molecular weight glutenin subunits (LMW-GS) were purified by this one-step procedure, whereas the remainder of the LMW-GS, comigrating in the acid system, were ...

Based on their mol. wts, notexin and scutoxin elute later than expected from gel filtration columns in multiple peaks [Francis et al. (1991) Toxicon 29, 85-96]. Notexins present in these peaks have identical amino acid sequences and unmodified amino acid side-chains. Scutoxin is an isoform of notexin which contains arginine ...

Earlier studies described the linkage of silver to antibodies using SH groups generated by the reduction of the SS groups using ascorbic acid (1) analogous to the Thakur and DeFulvio technique for linking technetium to antibodies. This work describes the linkage of silver to IgG after introducing SH groups by ...

Cajanus cajan lectin was isolated by ammonium sulfate fractionation and affinity chromatography on an IgM-Sepharose 6B column. Gel filtration and SDS-PAGE showed size homogeneity of the lectin. The lectin with M(r) 18,000 on SDS-PAGE had gel filtration behavior which was consistent with a molecular weight of 39 kDa and a ...

Whole desiccated venom of Bothrops pirajai was fractionated on a gel filtration (Sephadex G-75) column. Phospholipase A2, arginine esterase and clotting activity profiles of the six fractions (SI to SVI) obtained were determined. Fraction SIV from the gel filtration column was subjected to chromatography on SP-Sephadex C-25. It was resolved ...

The isolation and partial characterization of PAS-4 glycoprotein (78 kDa) from bovine milk fat globule membrane (MFGM) is described. PAS-4 was selectively extracted with Triton X-114 nonionic detergent and then fractionated on DEAE-Sepharose at pH 7.5. The PAS-4 fraction that was not bound on DEAE-Sepharose gave a single band by ...

Using human red blood cell ghost membranes, we have evaluated 5-nitro-2-[N-3-(4-azidophenyl)-propylamino]-benzoic acid and 5-nitro-2-[N-3-(4-azido-2,3,5,6-tetrafluorophenyl)-propylamino]- benzoic acid (FAzNPPB) as photoaffinity labeling agents based on the structure of the widely important Cl- channel blocker 5-nitro-2-(3-phenylpropyl-amino)-benzoic acid (NPPB). The tetrafluoro-substituted aryl azide was found to be a more effective photoinactivating agent than the ...

Two chitinases, designated PLC-A and PLC-B, were purified from the leaves of pokeweed (Phytolacca americana) using DEAE-cellulose column chromatography followed by gel filtration on Sephadex G-75, hydrophobic column chromatography, and ion-exchange FPLC. PLC-A and PLC-B are acidic and basic proteins having molecular masses of 25 and 29 kDa, and isoelectric ...

A new strain, Sphingomonas paucimobilis GS1, accumulated 6.5 g of a highly viscous exopolysaccharide per liter, using sucrose as a substrate. The anionic heteropolysaccharide contained the following, in grams per gram: glucose, 0.7; galacturonic acid, 0.11; glucuronic acid, 0.07; and acetate, 0.12. The viscosity of the exopolysaccharide (4.0 g/liter; 4,200 ...

This paper describes low-density mucus glycoconjugates released from feline trachea by dirhamnolipid (DRL), a toxin from Pseudomonas aeruginosa. Mucus glycoconjugates in feline tracheas were radiolabeled in vivo with 3H-proline and 14C-glucose. Control mucus and that released by 200 micrograms/ml DRL were dissolved in guanidine hydrochloride buffer (GuHCl) and chromatographed on ...

BACKGROUND: A topical formulation of lidocaine and prilocaine has been shown to provide effective anesthesia for certain superficial skin treatments. OBJECTIVE: To evaluate the efficacy of a topical self-occluding gel containing lidocaine 2.5% and prilocaine 3.5% in reducing the discomfort of 35% trichloroacetic acid (TCA) peels. METHODS: Ten patients who ...

A unique Cu,Zn-SOD was found and isolated from plaice Paralichthys olivaceus skin. Surprisingly, the properties of purified fish skin SOD were very different from those of SOD from other sources reported so far. The purified SOD was composed of four same subunits of 16 kDa and the molecular weight of ...

Linoleic acid is sequentially converted to 7S,8S-dihydroxy-9Z,12Z-octadecadienoic acid by the 8R-dioxygenase and hydroperoxide isomerase of the fungus Gaeumannomyces graminis, which is a common pathogen of wheat. The objective of this study was to separate and characterize the two enzyme activities. The isomerase activity was found mainly in the microsomal fraction ...

Many hormonal, metabolic and enzymatic alterations have been described in patients with chronic renal failure (CRF) after prolonged hemodialysis treatment. This toxicity has been associated with the accumulation of middle molecular weight components, called 'middle molecules', in their blood. To investigate the chemical nature of these molecules, 20 CRF patients ...

Bacillus pumilus PS213 isolated from bovine ruminal fluid was able to transform ferulic acid and p-coumaric acid to 4-vinylguaiacol and 4-vinylphenol, respectively, by nonoxidative decarboxylation. The enzyme responsible for this activity has been purified and characterized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of crude extract from a culture induced by ferulic ...

This study deals with the acid phosphatase (AcPase) of the Kurloff body (KB), a large (10 microns diameter) periodic acid-Schiff-positive lysosomal inclusion body present in Kurloff cells (KC). KC AcPase were extracted, with similar yields, either with non-ionic detergent solution or after Dounce homogenization in low ionic strength buffer suggesting ...

Dichelobacter nodosus, a Gram negative obligate anaerobe and causative organism of ovine footrot, secretes a family of extracellular acidic serine proteases with pI's in the range of 5.2 to 5.6, and a basic serine protease with a pI of approximately 9.5. Four acidic proteases (V1, V2, V3 and V5) from ...

The performance of acidic and basic urea polyacrylamide gels has been improved by adopting a two-part gel system with a concentration discontinuity to act as a stacking boundary and by increasing the urea concentration to 8 M. The contributions of primary amino-group and guanidino-group ionizations to mobility have been evaluated ...

We used an original potentiometric procedure to monitor the metabolic activity of biofilm-like structures and to assess their susceptibility to betalactam antibiotics. The reduction of exogeneous alpha-lipoic acid to dihydrolipoic acid by planktonic and agar-entrapped bacteria, metabolizing glucose, was followed by potential-time measurements using gold versus reference electrodes. Immobilized cultures ...

Comparative studies of the N-linked carbohydrate chains of human myeloma proteins of the IgA1 and IgA2 subclasses were performed. The N-linked carbohydrate chains were released by hydrazinolysis from the polypeptide backbone, converted to radioactive oligosaccharides by sodium borotritide reduction after N-acetylation and separated into one neutral and two acidic fractions ...

Two structurally distinct lectins were purified from the coelomic plasma of holothurian, Stichopus japonicus, by affinity chromatography on a porcine stomach mucin-conjugated agarose column, gel filtration on a Superose 6 column, and ion-exchange chromatography on a HiTrap Q-FPLC. The two lectins showed apparent molecular masses of about 400 kDa (SPL-1) ...

One form (form VI) of lysosomal arylamidase, having a molecular weight of 135,000 and an isoelectric point of 7.8, was purified from rat liver to apparent electrophoretic homogeneity. The enzyme consisted of three identical polypeptides with a molecular weight of 45,000, as judged by disk electrophoresis in the presence of ...

The synthesis of (2S,3S,4S)-4-[1-(4-azidobenzamidomethyl)ethenyl]-2-carboxy-3- pyrrolidineacetic acid (ABCPA) is described. This novel kainic acid analogue, bearing a photolabile functionality on the isopropenyl side chain, was proven to be a good inhibitor of [3H]CNQX and [3H]kainic acid binding on chick cerebellar membranes. [3H]ABCPA was photoaffinity cross-linked on the membrane fraction of chick ...

Polyacrylamide gel electrophoresis of fluorophore-labeled saccharides is a simple and sensitive separation method for glycan analysis. This method requires an aldehydic reducing carbon on the saccharide in order to react with the amine group of the fluorophore (8-aminonaphthalene-1,3,6-trisulfonic acid or 2-aminoacridone). We have used exoglycosidase-free ceramide glycanase from hard-shelled clam ...

Total glycans were isolated and purified from Lytechinus pictus embryos at early developmental stages by gel-filtration chromatography after pronase and DNase digestion, and alkali-borohydride treatment. Fractionation by Superose 6 and HPLC gel-filtration chromatography revealed three major glycan fractions of 580, 150 and 2 kDa consistently throughout development up to the ...

We have previously shown that mouse sarcoma 180 cells produce vascular endothelial growth factor [VEGF; Rosenthal et al., 1990, Growth Factors, 4: 53-59], an endothelial mitogen that stimulates angiogenesis. Recent reports have implicated metalloproteinases and their inhibitors in the regulation of vascular morphogenesis, tumor invasion, and metastasis. We report here ...

The heparin-binding lectin, Anadarin MS, from the plasma of the marine clam Anadara granosa is purified through affinity chromatography on heparin-Sepharose 4B followed by gel filtration on a Sepharose 6B column. The purified lectin is a pentameric protein of native M(r) 300 kDa and is composed of identical subunits of ...

The synthesis of methanol insoluble-[14C]labeled-polymers from [14C]sucrose by S. mutans glucosyltransferases was stimulated up to four-fold by the presence of highly purified, lipoteichoic acid (LTA) from S. sanguis. Gel filtration chromatography (Sepharose 4B) and ion exchange chromatography (DEAE-Bio-Gel A) of the [14C]labeled-polymers formed in the presence of [3H]glycerol-labeled-LTA showed that ...

N-Carboxymethylchitosan from crab and shrimp chitosans was obtained in water-soluble form by proper selection of the reactant ratio, i.e. using equimolar quantities of glyoxylic acid and amino groups. HPLC determinations of glyoxylic and glycolic acids, in conjunction with NMR analysis, permitted identification of the structure of the product, which is ...

The isolation of human serum transferrin (Tf) using recycling isotachophoresis (RITP) and recycling isoelectric focusing (RIEF) with simple buffers is described. Serum fractionation, the first step in the protocol for Tf purification, is shown to be easily performed either by RITP of filtered serum using low molecular mass spacers or ...

The egg shell of Hymenolepis diminuta is composed of 31.7% protein and 2.9% carbohydrate (by weight), as determined using the Lowry and phenol-sulfuric acid assays and bovine serum albumin and glucose, respectively, as standards. Amino acid analyses of egg shells demonstrated the presence of 16 naturally occurring amino acids and ...

An abnormal fibrinogen was discovered in a clinically asymptomatic woman from Italy. Routine coagulation studies revealed prolonged thrombin and reptilase clotting times and a discrepancy between the plasma fibrinogen levels determined by the clotting assay and electroimmunoassay. Release of fibrinopeptides A and B from fibrinogen Milano V by thrombin was ...

A new bacteriocin, gassericin A, was purified from the culture fluid of Lactobacillus gasseri LA39 mainly by reverse-phase (RP) chromatography. The purification of gassericin A from a modified MRS broth, in which Tween 80 had been replaced by oleic acid, resulted in a 4500-fold increase in specific activity with a ...

A method is described for the electrophoretic analysis of fluorophore-labeled acidic glycans. Various glycoproteins were treated with the enzyme, peptide N-glycosidase F, to release mixtures of asparagine-linked glycans that were then labeled with the fluorophore 2-aminoacridone, and the resulting derivatives electrophoresed in high-density polyacrylamide gels. The acidic glycans were separated ...

A lysophosphatidic acid (LPA)-hydrolysing lysophospholipase was purified from rat brain and characterized. This membrane-bound lysophospholipase was solubilized by using n-octyl glucoside and purified by sequential cation, hydrophobic and gel-filtration chromatography. The purified protein has a mass of 80 kDa as assayed by SDS/PAGE. This lysophospholipase catalysed the hydrolysis of a ...

Pseudomonas strain BL072 produces an acylase enzyme active in hydrolyzing glutaryl-7-aminocephalosporanic acid to 7-aminocephalosporanic acid. This acylase was purified by column chromatography and gel electrophoresis. The native acylase was composed of two subunits of approximately 65 and 24 kDa, though some heterogeneity was seen in both the native acylase and ...

The physiologically relevant stress of a flux of H2O2 increased hemoglobin (Hb) degradation in red blood cells (RBC) and increased the proteolytic susceptibility of Hb in vitro. After exposure to low H2O2 flux rates (6-32 microM/min) Hb exhibited increased exposure of hydrophobic (Trp, Met) and basic (Lys) amino acid R ...

Purification of sialyl-Le(a)-carrying mucins from primary human bile by trichloroacetic acid precipitation, delipidation, and gel filtration in guanidinium chloride gave three separable fractions, one of which was further purified by affinity chromatography. These fractions, named SBG1 (for soluble bile glycoprotein), SBG2, and SBG3 had molecular masses of > 1100, 800-950, ...