Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0272170 (SDS)
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The aim of this study was to show psychopathological and especially psychophysiological data of depressed inpatients which were collected during a 28-day double-blind study with 68 depressed inpatients given clomipramine 150 mgr/die) or maprotiline/oxaprotiline 150 mgr/die). Using Hamilton-Depression-Scale, Self-depression-scale (SDS by Zung), Beschwerdeliste (BL) and Befindlichkeitsskala (Bf-S) by v. Zerssen an impressive reduction of depression scores was observed in all rating scales with some superiority of clomipramine. But at the end of the study at treatment day 28 both groups - clomipramine and maprotiline/oxaprotilin - had similar results. During treatment each patient was part of a weekly habituation experiment using electrodermal activity in an orientation reaction as measure. Contrary to other studies (Lader 1975 or Heimann 1976, 1980) there was no difference in the course of psychophysiological data (EDA) between both treatment groups or between depressive agitation and psychomotor retardation. There was also no relationship between reactivity in EDA at study begin and treatment results.
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PMID:[Psychopathologic and psychophysiologic follow-up of inpatient depressed patients with standardized treatment with clomipramine and oxaprotiline]. 171 52

Incubation of lysates prepared from amebocytes of the horseshoe crab (Limulus polyphemus) with bacterial endotoxin results in coagulation and formation of a solid gel. Although Limulus gels remained solid indefinitely, if undisturbed, they were easily disrupted by mechanical agitation. Chemical solubility studies of gelled lysates demonstrated rapid solubilization of gels in monochloroacetic acid, a property of clots that have not been covalently stabilized; but in contrast demonstrated resistance to solubilization by urea, a property of stabilized clots. Analysis of solubilized proteins by polyacrylamide gel electrophoresis in SDS demonstrated coagulin, the designation for the activated form of coagulogen (the clottable protein) that forms a gel, only in samples derived from clotted lysate that had been previously incubated with monochloroacetic acid, but not in samples following incubation with urea, confirming the results of the chemical solubility studies. Enzymatic assays for transpeptidase (Factor XIII-like) activity in either native or gelled Limulus lysates were negative. Furthermore, analysis for covalently crosslinked peptides in gelled coagulin confirmed the absence of intermolecular gamma-glutamyl-epsilon-lysyl bonds. Therefore, the stable gels formed following coagulation of Limulus lysate by bacterial endotoxin are not covalently crosslinked.
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PMID:Stability of gels formed following coagulation of Limulus amebocyte lysate: lack of covalent crosslinking of coagulin. 278 19

Adherence of Porphyromonas gingivalis to early plaque bacteria, such as Streptococcus gordonii, is considered an important colonization mechanism. The molecules that mediate this interspecies binding have not been determined. Fimbriae were prepared from P. gingivalis 33277 by mild agitation, ammonium sulfate precipitation and DEAE-Sepharose chromatography. In a nitrocellulose blot adherence assay, purified fimbriae inhibited S. gordonii G9B-P. gingivalis 33277 binding by up to 54%. In addition, fimbriae bound to S. gordonii cells in a dot-blot assay. Incubation of fimbriae with S. gordonii cells followed by washing, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), electroblotting and probing with P. gingivalis antibodies also revealed that the fimbriae bind to S. gordonii. In contrast, S. gordonii did not interact with fimbriae that were first subjected to SDS-PAGE and electroblotting or deposited on a nitrocellulose membrane, suggesting that conformational determinants of the fimbriae may be important in binding. The results indicate that binding between P. gingivalis and S. gordonii is mediated, at least in part, by the porphyromonads' fimbriae.
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PMID:Involvement of Porphyromonas gingivalis fimbriae in adherence to Streptococcus gordonii. 790 42

Optimized acetyl esterase enzyme production conditions using Aspergillus niger ATCC 10864 in 14-L fermentation jars were determined to be 33 degrees C, 1.5 vvm aeration, and 300 rpm agitation without pH control. The acetyl esterase was purified by precipitation in 60-80% saturation in ammonium sulfate. The pellet was applied directly to a Pharmacia high-load Phenyl Sepharose column for hydrophobic interaction chromatography and purified to homogeneity in two steps. Stability and kinetic characteristics of the acetyl esterase were determined over a pH range of 4.0-7.5 and from 4 to 45 degrees C. At temperatures > 25 degrees C, stability was superior at pH values < 5.0. The temperature activity optimum was 35 degrees C, and the pH optimum was 7.0. The Vmax was determined to be 46,700 U/mg protein, and the Km was 0.023M p-nitrophenyl acetate at pH 6.5 in 0.2M phosphate buffer at 35 degrees C. The mol wt of the enzyme was 35,000 dalton by size-exclusion chromatography and SDS gel electrophoresis. The N-terminal amino acid sequence and the glycosylation composition were also determined.
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PMID:Purification and characterization of an acetyl esterase from Aspergillus niger. 801 Jul 67

Three denaturing techniques have been evaluated for their ability to induce irreversible aggregation and precipitation of recombinant porcine growth hormone (pGH). The denaturing stimuli included thermal denaturation, interfacial denaturation through the introduction of a high air/water interface by vortex agitation, and a guanidine (Gdn) HCl technique which involved rapid dilution of a partially unfolded state of pGH to nondenaturing conditions. Soluble and insoluble pGH fractions were evaluated for the presence of covalently modified species and soluble aggregates by size exclusion chromatography (SEC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and isoelectric focusing (IEF). In each of the three denaturation methods, precipitation was found to be irreversible, as the precipitated pellet could not be solubilized upon resuspending in buffer. The soluble pGH fractions consisted of only monomeric material and the insoluble protein pellet could be completely solubilized with Gdn HCl or SDS. There was no evidence of detectable covalent modifications in the precipitated protein pellet following any of the three denaturation techniques. Three excipients, Tween 20, hydroxypropyl-beta-cyclodextrin (HPCD), and sorbitol were evaluated for their stabilizing ability using each of the three denaturation methods and the degree of stabilization was found to be dependent upon the denaturing stimulus incorporated. Tween 20 was found to be highly effective in preventing pGH precipitation using the interfacial and Gdn techniques and was moderately effective using the thermal denaturation method. Inclusion of HPCD in the sample buffer significantly reduced precipitation using the thermal and interfacial methods but was ineffective in the Gdn technique.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Techniques for assessing the effects of pharmaceutical excipients on the aggregation of porcine growth hormone. 837 57

Aeromonas sp., grown in tryptone soya broth supplemented with yeast extract, 0.6%, pH 7.5, and incubated with agitation at 100 oscillations/min for 15 h at 37 degrees C produced optimal amounts of beta-haemolysin and cytotonic enterotoxin. More prolonged incubation resulted in the loss of enterotoxic activity and anion exchange chromatographic analysis indicated the presence of a moiety capable of breaking down the toxin. Anion exchange fast protein liquid chromatography resulted in a single peak of haemolytic activity and two peaks with enterotoxic activity. The cytotonic enterotoxin was purified from the fraction most active in the infant mouse assay; the second peak, which did not cross-react immunologically, may represent a second cytotonic enterotoxin. Neither peak was observed in the chromatographic fractions of filtrates from strains devoid of activity in the infant mouse assay. Purified enterotoxin, estimated to have a mol. wt of 15 kDa by SDS-PAGE, caused fluid accumulation in the infant mouse assay, was non-haemolytic to rabbit erythrocytes, caused an increase in cAMP activity in tissue culture cells and did not cross-react immunologically with components of cholera toxin or the whole toxin. Purified beta-haemolysin had an estimated mol. wt of 55 kDa, lysed rabbit erythrocytes and did not cause fluid accumulation in the infant mouse test.
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PMID:Isolation and purification of Aeromonas sobria cytotonic enterotoxin and beta-haemolysin. 838 63

Aspergillus fumigatus and other species of the same genus can cause different pathological processes amongst which can be found pulmonary hypersensitivity, intracavitary colonization and invasive and disseminated aspergillosis. The diagnosis of these processes is helped in a great way by the immunological response of the patient to Aspergillus antigens or by the detection of these as a metabolic product in blood and other fluids such as urine. To date, no firm agreement exists regarding the best preparation and standardization technique of aspergillar antigens, which is however the most important step towards the development of immunological diagnostic tests. In this study, two methods for the obtention of A. fumigatus somatic antigens have been compared, using 3 strains of different origins. The main differences between both antigenic extraction methods were the liquid media used for cultures (Czapek and Glucose peptone), the incubation times (4 weeks and 3 days respectively), the type of agitation during the incubation period (intermittent or continuous) and the dialysis and concentration method. The antigens obtained were standardized by the determination of the proteic and hydrocarbonate content, vertical electrophoresis in SDS PAGE, immunoprecipitation and immunoblotting against experimental antisera. From the comparison between both extraction methods, it was found that the highest proteic content was obtained with the somatic antigens of mycelium developed in Czapek for 4 weeks, whereas fungi cultivated in Glucose peptone had a higher glucide content. Vertical electrophoresis revealed the presence of 6 common fractions in all the antigens, whilst some fractions were in majority, only found in one antigen such as the MW 109 kDa, present in the GP 69 antigen.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Aspergillus fumigatus somatic antigens: comparison between two methods of preparation and immunochemical characterization. 855 91

The present study reports on a 17-year-old male who ingested approximately 70 ml trichloroethene (TRI) in a suicide attempt. The patient developed fever, tremor, general motor restlessness, and sinus tachycardia and lost consciousness 5 h after poisoning. After 5 days of intubation under narcosis with forced hyperventilation and diuresis he regained consciousness. During this period blood and urine were collected and TRI and its metabolites were quantified. The highest concentration of TRI in blood was detected 13 h after ingestion. Trichloroethanol and trichloroacetic acid, metabolites of the cytochrome P450-mediated pathway, and N-acetyl-S-(1, 2-dichlorovinyl)-l-cysteine and N-acetyl-S-(2, 2-dichlorovinyl)-l-cysteine from the glutathione-dependent pathway of TRI were quantified in urine samples. Besides these known metabolites in humans, chloroacetic acid and dichloroacetic acid were identified for the first time in urine of a human exposed to TRI. Although the patient exhibited normal levels of glucose and total protein in urine, excretion of alpha1- and beta2-microglobulin as well as beta-NAG was significantly increased. In addition to these typical markers of selective tubule damage, analysis of the urinary protein pattern by SDS-PAGE revealed increased excretion of several low-molecular-mass proteins between 10,000 and 50,000 Da, clearly indicating tubular damage. Based on the elucidated glutathione-dependent mechanism for the nephrotoxicity of TRI, activation of the formed S-conjugates by beta-lyases to reactive intermediates may account for the observed renal effects after a single, high dose of TRI.
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PMID:Acute intoxication with trichloroethene: clinical symptoms, toxicokinetics, metabolism, and development of biochemical parameters for renal damage. 952 Mar 51

This work evaluates the efficacy of the spray-drying technique to prepare poly(epsilon-caprolactone) (PCL) microparticles containing an entrapped model antigen (bovine albumin, BSA). The presence of a stabiliser was found to be an important parameter when preparing PCL microparticles containing a hydrophilic antigen. The effect of various technological parameters (concentration of the polymer and protein solutions, organic/aqueous phases ratio, nature of solvents and emulsion parameters such as duration and speed of agitation) on microparticle morphology and size, BSA entrapment and encapsulation efficiency was studied. Microparticles were characterized by a mean size from 9.56+/-0.25 to 24.31+/-2.87 microm and a BSA entrapment from 0.80+/-0.02 to 24.21+/-0.23% (w/w). SDS-PAGE electrophoresis and isoelectric focusing (IEF) confirmed the conservation of the physicochemical characteristics of the BSA entrapped within PCL microparticles produced by spray-drying. Together, these results showed that spray-drying is an efficient technique to overcome the key obstacle that represents the scaling-up of the manufacturing process to produce sufficient quantities of vaccine for clinical trials and, ultimately, commercialization.
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PMID:Influence of various technological parameters on the preparation of spray-dried poly(epsilon-caprolactone) microparticles containing a model antigen. 1089 88

In a 5L fermentor the production conditions of alkaline protease gene engineering strain BA071 were investigated. The maximum activity of alkaline protease reached 24,480 u/mL in 40 hours of fermentation by combination of enhancing aeration and changing the agitation rate. The fast purification method of recombinant protease was conducted with FPLC (Fast Protein Liquid Chromatography). The crude enzyme, treated with ammonium sulfate fractionation and decolored with DEAE-A-50 and polyethylene glycol concentration, was purified with CM-Sephadex-C-50 and Sephadex-G-75. The purified enzyme appears homologous on SDS-PAGE. The purity of enzyme was increased 76.2 times. SDS-PAGE analysis showed that the molecular weights of expressed recombinant products were about 28 kD. The optimal reaction pH and temperature of recombinant enzyme were at pH11 and 60 degrees C, respectively. The recombinant enzyme exhibited high temperature tolerance and was stable at a wide range of pH. Ca2+, MG2+ can enhance the stability of the recombinant enzyme. While the protease activity of the enzyme was strongly inhibited by Hg2+, Ag+, PMFS [symbol: see text] DFP, and was not affected by SDS and Urea.
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PMID:[Study on fermentation condition of alkaline protease gene engineering strain and the purification and characterization of recombinant enzyme]. 1267 45


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