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Query: UMLS:C0040822 (tremor)
 18,428 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Propagules of Rhizoctonia solani grown in modified Czapek's medium containing sodium polypectate or carboxymethyl cellulose as a sole carbon source produced both extracellular and cell-bound polygalacturonase (PG), and cellulase (Cx), respectively. The cell-bound enzymes can be released to various extents by shaking the germinating propagules in solutions of NaCl, KCl, phosphate buffer, Na2EDTA (ethylenediaminetetraacetate), detergents such as Triton X-100 (octyl phenoxypolyethoxyethanol), Tween 80 (polyoxyethylene sorbitan monooleate), Celmusol, and distilled water. Sodium dodecyl sulfate (SDS) inactivated both PG and Cx but did no affect Cx activity in phosphate buffer solution. PG was more easily released by salts from the mycelium of R. solani than Cx. The release of both enzymes was a passive process and was not due to an osmotic effect. The amount of the cell-bound fraction was correlated with the total amount of the extracellular fraction rather than with the mycelial growth. At least one-third of the cell-bound fractions of both enzymes was found to be associated with the cell wall fraction of the mycelium.
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PMID:Release of cell-bound polygalacturonase and cellulase from mycelium of Rhizoctonia solani. 80 41

We have recently characterized a bradykinin (BK) receptor in rat renal mesangial cells (1). Activation of this receptor is associated with PGE2 release and IP3 formation suggesting involvement in cell contraction which can be linked to the control of the glomerular filtration rate (2). Whether this mesangial BK receptor is the unique glomerular BK receptor remains to be elucidated. In an attempt to answer to this question, we performed binding studies using decapsulated isolated glomeruli. Scatchard analysis of the binding data obtained with this preparation revealed the presence of two distinct B2-kinin binding sites. However, a consistent difference was observed in both the affinity and the density. We further investigated the pharmacological binding profile after an initial step of solubilization. Several experiments were performed to establish optimal conditions of solubilization. For this, different detergents such as Triton X-100, CHAPS and n-octyl beta-D glucopyranoside were tested at various concentrations, durations and temperatures of incubation. The binding was performed with two different [125I]-Tyr0-BK concentrations (0.5 and 7 nM) with either untreated decapsulated glomeruli or solubilized preparation for 45 minutes at +4 degrees C in the binding buffer containing a mixture of protease inhibitors. The greatest binding was achieved after treating glomeruli with 25 mM n-octyl beta-D glucopyranoside for 60 minutes at +4 degrees C under constant shaking. Two B2-kinin receptors of different affinities were detected. The same binding characteristics were obtained both in the 12,000 x g and 100,000 x g supernatant.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Characterization of two different affinity B2-kinin binding sites in rat glomeruli. 133 39

A simple, rapid SDS-Page method for protein profiles analysis of Actinomyces species was developed. Eighteen strains (12 reference strains and 6 fresh isolates) representing four species were used in this study. Eight detergents were tested for protein extraction. Cell extracts were obtained by shaking the bacteria suspended in a solution containing a detergent and glass beads. After electrophoresis, according to the Laemmli's technique, two protein stain procedure were tested (Coomassie blue and silver stain). Best results were obtained with 0.5% Triton X-100 for 4 h and the silver stain procedure of Oakley. Protein profiles analysis showed that the method is reproducible and distinguishes not only species but sometimes also subspecies.
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PMID:Improved SDS-Page method for protein profiles analysis of actinomyces species. 246 59

The acrosomal matrix of hamster spermatozoa was enriched and characterized. Acrosomal matrices were released from spermatozoa with shaking in a pH 5.2 buffer containing Triton X-100 and protease inhibitors, and enriched on a glass-bead column. Phase-contrast microscopy indicated that 70-80% of the acrosomal matrices were released from the spermatozoa and only minor contamination from sperm heads was detected. Transmission electron microscopy confirmed the low level of contamination in the preparation and revealed a bilaminar structure similar but not identical to that of guinea-pig acrosomal matrix. One- and two-dimensional SDS-polyacrylamide gel electrophoresis (SDS-PAGE) showed the acrosomal matrix to be a complex structure enriched for several polypeptides. Proteinase activity was demonstrated by gelatin-SDS-PAGE. The major activity corresponded to bands of relative molecular masses (Mr) of 56,000, 51,000 and 48,000 with two minor bands of Mr 30,000 and 28,000. The lectin Pisum sativum agglutinin (PSA) bound to the anterior head of spermatozoa and isolated acrosomal matrix as judged by fluorescence microscopy using FITC-PSA. Western blots of spermatozoa and acrosomal matrices followed by overlay with biotinylated PSA indicated that there are at least two PSA-binding glycoproteins of Mr 60,000 and 72,000.
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PMID:Characterization of isolated acrosomal matrices from hamster spermatozoa. 329 98

The effects of eluent composition, pH, and chaotropic agents on the recovery of T2, MS2, and indigenous coliphages from various foods were investigated. Additionally, methods of sample suspension and clarification were evaluated for coliphage recovery and application to various foods. Clarified sample suspensions were assayed for coliphages with a modified agar layer technique and appropriate Escherichia coli hosts. Centrifugation and polypropylene mesh filtration were more rapid and effective than glass wool filtration for clarification of sample suspensions and subsequent recovery of coliphages. Blending, stomaching, and shaking procedures were generally comparable for sample liquefaction and release of coliphages from foods. Complex basal eluents, EC medium and 1% casein, were generally more effective than a less complex eluent, phosphate buffer, for elution of coliphages from foods. For most foods, incorporation of sodium chloride or chaotropic agents, i.e., sodium trichloroacetate, urea, Tween 80, Triton X-100, and sodium nitrate, into basal eluents did not enhance recovery of coliphages. Indigenous coliphage recovery was not affected by sample suspension pH over a range of 6.0 to 9.0. With an optimal procedure, i.e., EC medium eluent, blending, and centrifugation, the recovery of T2 and MS2 ranged from 48 to 81% and from 58 to 100%, respectively, depending on the food type.
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PMID:Methodology for enumeration of coliphages in foods. 352 56

Our recent morphological studies showed that basement membranes isolated from renal tubules tended to collapse and form folded sheets while glomerular basement membranes were more resilient. In an effort to study the shapes of various isolated basement membranes in undissociated tissues, a method was developed to remove all cellular elements and leave the extracellular matrix and associated basement membranes intact. Accordingly, transplant quality human kidneys were harvested, perfused with Collin's medium and transported to the laboratory on ice. The renal cortex was then peeled away by blunt dissection, further minced to 2 mm3 and placed in 1 mM EDTA (with gentle intermittent stirring) for 72 h at 4 degrees C. Solubilization of cellular materials was carried out by successive washings with 3% Triton X-100, 0.025% DNAse and 1-4% sodium deoxycholate (all with gentle stirring or shaking at 22 degrees C). Each solution contained 0.1% sodium azide. At the level of fine structure, glomerular, tubular, Bowman's capsular and peritubular capillary basement membranes all maintained their respective shapes and did not collapse. Glomerular basement membrane was particularly striking in this regard and exhibited an open, lobulated form that closely resembled its in vivo histoarchitecture. Moreover, when the acellular tissue blocks were prepared for scanning electron microscopic observation, the glomerular basement membranes exposed at the surface of the block showed a remarkable structural rigidity. These basement membranes were free-standing, convoluted electron-dense sheets, continuous with highly folded central mesangial regions. It seems significant that glomerular basement membranes maintain their in vivo conformation irrespective of the presence of other extracellular matrix components while removal of these materials by organ subfractionation results in folding and general shapelessness of tubular basement membrane. It is possible that in addition to its unique role in filtration, glomerular basement membrane may also serve to preserve glomerular shape, regardless of changing cell populations or alterations in hydrostatic pressures.
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PMID:Preparation and histoarchitecture of ultrastructurally pure glomerular basement membrane. 703 91

A detergent scrub technique using wash fluid consisting of 0.075 M phosphate-buffered saline, pH 7.9 containing 0.1 per cent Triton X-100 was evaluated for the quantitative culture of Malassezia pachydermatis from canine skin. Preliminary studies showed that the detergents Triton X-100, Tween 40 and Tween 80 were equally able to disperse suspensions of pure cultures of M pachydermatis, but that the yeast counts were significantly reduced (P < 0.001) after suspension in saline, Triton X-100 or Tween 40 for two hours. The counts in skin washings were also reduced (P < 0.001) after suspension for three hours in 0.1 and 0.05 per cent solutions of Triton X-100. Vortexing, or manual or mechanical shaking of the samples yielded comparable counts. The correlation between the counts on diseased skin measured by using detergent scrubs and a contact plate technique was highly significant (P < 0.001). The detergent scrub technique was suitable for the quantitation of M pachydermatis on canine skin provided that the samples were processed without delay.
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PMID:Evaluation of a detergent scrub technique for the quantitative culture of Malassezia pachydermatis from canine skin. 776 91

1. Immunocytochemical and biochemical techniques have been used to localize and characterize a novel plasma membrane-associated, neutral-pH-optimum alpha-L-fucosidase from rat spermatozoa. Light and electron microscopy specifically localized the fucosidase on the plasma membrane of the convex region of the principal segment of testicular and cauda epididymal sperm heads. Immunoreactivity for alpha-L-fucosidase was also detected in the Golgi apparatus of spermatocytes and spermatids but no immunoreactivity was observed in the acrosome. 2. Fractionation of epididymal sperm homogenates indicated that over 90% of the alpha-L-fucosidase activity was associated with the 48,000 g pellet. This pellet-associated activity could be solubilized with 0.5 M NaCl but not with 0.5% Triton X-100, suggesting that fucosidase is peripherally associated with membranes. Sucrose-density-gradient centrifugation of sperm homogenates indicated that fucosidase was enriched in the plasma membrane-enriched fraction. Analysis of alpha-L-fucosidase on intact epididymal sperm indicated that the enzyme was active, displayed linear kinetics and had a pH-activity curve (with an optimum near 7) which was comparable to that of fucosidase from epididymal sperm extracts. These results further suggest that fucosidase is associated with plasma membranes, and that its active site is accessible to fucoconjugates. Evidence that most of the fucosidase is associated with the exterior of the plasma membrane came from studies in which intact sperm had fucosidase activity comparable to that of sperm sonicates, and from studies in which approx. 90% of the fucosidase activity on intact sperm could be released from the sperm by gentle shaking with 0.5 M NaCl. Isoelectric focusing indicated that the NaCl-solubilized epididymal sperm fucosidase appears to have one major and one minor isoform with pIs near 7.2 and 5.2, respectively. SDS/PAGE and Western blotting indicated that the NaCl-solubilized extract of epididymal sperm contains two protein bands of 54 and 50 kDa which were highly immunoreactive with the IgG fraction of anti-fucosidase antibodies. Although the function of the novel sperm fucosidase is not known, its specific localization to the plasma membrane of the region of the rat sperm head involved in sperm-egg binding and its high enzymic activity at neutral pH on intact sperm suggest that this enzyme may have a role in sperm-egg interactions.
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PMID:Immunocytochemical localization and biochemical characterization of a novel plasma membrane-associated, neutral pH optimum alpha-L-fucosidase from rat testis and epididymal spermatozoa. 883 25

The ability to produce cellulose and xylan degrading enzymes by different strains of Thecotheus pelletieri, in liquid synthetic media with cellulose and xylan as inducers, was compared. All the strains tested were able to grow and produce cellulases and xylanases, being the strain BAFC 2077 the best producer. Several cultural conditions were analysed in order to optimise enzyme production by strain 2077. Shaking cultures gave higher yields of cellulases and xylanases compared with stationary ones. Asparagine at 0.75 g N/L was the best nitrogen source in promoting enzyme production. The influence of different surfactants on enzyme production was studied. Tween 80 exhibited no effect on growth and enzyme production, whereas Tween 20 and Triton X-100 were inhibitory. By means of studies of variation of cellulose/xylan ratio in the culture medium we determined that cellulose and xylan induced cellulase and xylanase synthesis, being the specific substrates the most effective. The inducible behavior of cellulases and xylanases in T. pelletieri was determined by means of inhibition of protein synthesis by cycloheximide and ethidium bromide. Moreover, we found that glucose as well as xylose repressed cellulase and xylanase synthesis in T. pelletieri.
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PMID:Cellulose and xylan degrading enzymes in Thecotheus pelletieri. 1114 50

Cultivation conditions affecting feather degradation by Bacillus sp. FK 46 were investigated. The results showed that feather was almost completely degraded under the following conditions: 1% whole chicken feather as a substrate at the initial medium pH of 9 with 5% bacterial inoculum, at a temperature of 37 degrees C and a shaking speed of 250 rev/min. Glucose, methanol, Tween 80 and Triton X-100, however, had no effect on feather degradation. After feather was degraded, its residue and fermented broth would become a protein feed for animals.
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PMID:Feather degradation by Bacillus sp. FK 46 in submerged cultivation. 1268 66


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