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Fluorescein-labeled beta-glucosidase as a bacterial stain.

Fluorescein isothiocyanate-labeled beta-glucosidase was used as a simple staining reagent with selected gram-positive and gram-negative organisms. Staining in situ appeared to be dependent on the presence of accessible glycosidic-type linkages in the bacterial cell wall. Extensive wall damage or lysis did not occur when stained cells were suspended in washing and mounting solutions. The apparent specificity of labeled enzyme for wall substance was tested by blocking reactions, staining of isolated cell walls, and failure to stain substances lacking appropriate glycosidic linkages. Severe cell wall lesions were produced after prolonged contact with labeled enzyme, and this phenomenon may also be related to staining specificity. Gram-negative organisms and spores were poorly stained unless protected glycopeptide substrate was previously exposed by treatment of cells with thioglycolic acid or dilute alkaline sodium hypochlorite solution. A potential for staining tissues and cell lines may also exist. Some possible applications of labeled enzymes are briefly discussed.

Bacillus anthracis↗

EFFECT OF CHELATING AGENTS ON THE GROWTH OF ESCHERICHIA COLI IN SEAWATER.

Jones, Galen E. (Scripps Institution of Oceanography, University of California, La Jolla). Effect of chelating agents on the growth of Escherichia coli in seawater. J. Bacteriol. 87:483-499. 1964.-Escherichia coli did not grow at 37 C, or grew only after a prolonged lag phase in filter-sterilized basal seawater medium (synthetic or natural seawater supplemented with glucose, NH(4)Cl, and K(2)HPO(4)). When this basal medium was enriched with 0.01% or less organic matter, such as casein hydrolysate, peptone, or yeast extract, growth always occurred after a short lag phase. Adding 10(-5)m cysteine or autoclaving the seawater gave a similar effect. A variety of organic chelating agents (histidine, glycine, methionine, glycylglycine, 8-hydroxyquinoline, thioglycolic acid, o-phenanthroline, disodium ethylenediaminetetraacetic acid, etc.) reversed the toxicity of filter-sterilized basal seawater medium in concentrations predictable from stability constants. Even metal-complexing agents such as Na(2)S(2)O(3), Na(2)S, and NaCN in appropriate concentrations reversed toxicity. The quality of the distilled water and the treatment of glassware had a significant effect on the growth of E. coli in basal seawater medium. It was concluded that iodate is probably not the toxic substance for E. coli in seawater, since relatively high concentrations were stimulatory. The inhibition resulting from the individual salts of synthetic seawater was proportional to their concentration; NaCl was most inhibitory. This toxicity is believed to be derived from trace impurities in the reagent-grade chemicals used to prepare synthetic seawater. Evidence was also found for the toxicity of heavy metals in natural seawater. Heavy metals in seawater appear to inhibit growth but not respiration.

Chelating Agents↗

In vitro thrombogenicity investigation of new water-dispersible polyurethane anionomers bearing carboxylate groups.

New segmented polyurethane (PU) anionomers based on hydroxytelechelic polybutadiene were synthesized via an aqueous dispersion process. Incorporation of carboxylic groups was achieved using thioacids of different length. Surface properties were investigated by mean of water absorption analysis and static contact-angle measurements using water, diiodomethane, formamide and ethylene glycol. Blood compatibility of the PUs was evaluated by in vitro adhesion assays using 111In-radiolabeled platelet-rich plasma and [125I]fibrinogen. Morphology of the adhered platelets was examined by scanning electron microscopy (SEM). Results were compared to two biomedical-grade PUs, namely Pellethane and Tecoflex. Insertion of carboxylic groups increased surface hydrophilicity and limited water uptake ( < 8% for an ion content of 5% by weight). Surface energy of all synthesized PUs was between 40 and 45 mJ/m2. Platelet adhesion and fibrinogen adsorption on the PU anionomer surfaces were affected as a function to the increase of graft length; thiopropionic was the most haemocompatible, followed by thiosuccinic and then thioglycolic acid. SEM analyses of all ionic PU samples exhibited low platelet adhesion to surfaces with no morphological modification. In conclusion, increased hydrophily, dynamic mobility and charge repulsion are synergistic key factors for enhanced haemocompatibility.

Absorption↗

High-grade intensification of the end-product of the diaminobenzidine reaction for peroxidase histochemistry.

A simple and reliable method is described for the intensification of the end-product of the diaminobenzidine reaction demonstrating peroxidase activity. After completing the diaminobenzidine reaction, the preparations to be intensified are immersed first in thioglycolic acid solution, then in distilled water, and finally in a special physical developer containing silver nitrate.

3,3'-Diaminobenzidine↗

Silver-enhanced diaminobenzidine-sulfide (SEDS): a technique for high-resolution immunoelectron microscopy demonstrated with monoamine immunoreactivity in monkey cerebral cortex and caudate.

A common frustration of immunoelectron microscopy (IEM) is the density of the 3,3'-diaminobenzidine (DAB) label, which obscures intracellular details of labeled structures. To overcome this problem, a silver enhancement protocol was developed which leaves silver deposits on very low levels of DAB. The resulting label is composed of easily visualized punctate silver deposits, localized in processes with little or no detectable DAB. This technique incorporates several modifications into previously described methods for silver enhancement of DAB. The principal innovation is to pretreat the DAB label with sodium sulfide before silver enhancement, which substantially increases the sensitivity of the silver enhancement. In addition, cysteine was used in place of thioglycolic acid to suppress tissue argyrophilia, allowing use of both glutaraldehyde- and paraformaldehyde-fixed tissue without degradation of ultrastructure. We demonstrate this technique with dopamine, norepinephrine (NE), and serotonin (5HT) immunoreactivity in monkey prefrontal cerebral cortex and with dopamine immunoreactivity in the anterior caudate. The punctate label allows essentially unobscured visualization of the intracellular details and cell membranes of these monoamine axons. Whereas 5HT axons formed small asymmetric synapses, dopamine and NE axons typically formed small symmetric synapses with notably subtle membrane specializations. It is likely that these are often obscured by conventional DAB labeling. The use of several preparations indicates that this technique will be useful with a variety of antibodies. It might also provide an attractive alternative to colloidal gold, especially with glutaraldehyde-fixed tissue which is not easily penetrated by gold particles.

3,3'-Diaminobenzidine↗

Isolation of a hepatic iodothyronine 5'-monodeiodinase by nondenaturing agarose gel electrophoresis.

We have examined the application of nondenaturing agarose gel electrophoresis for isolation of the catalytically active iodothyronine 5'-monodeiodinase (5'-MD) in rat liver microsomes. Preliminary studies showed that most ingredients of conventional sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, including acrylamide, SDS, and bromophenol blue, markedly inhibited 5'-MD activity in solubilized rat liver microsomal proteins (SRLMP). We replaced these inhibitory components with those that had little or no inhibitory effect on the 5'-MD and devised conditions for nondenaturing agarose gel electrophoresis for isolation of a 5'-MD protein. SRLMP (up to 120 micrograms protein/well), prepared by preincubation with 5 mM 3-[(3-cholamidopropyl)dimethylammonio 1-propanesulfonate] (CHAPS) and 1% 2-mercaptoethanol, were subjected to electrophoresis in 0.35% agarose gel in 20 mM Tris-HCl, 10 mM EDTA, pH 6.6 buffer containing 2 mM CHAPS and 1 mM thioglycolic acid. Electrophoresis was carried out for 14 h at 4 C using 2.5 V/cm. After phoresis, the gel was sliced into 16 fractions, and homogenates of each fraction were tested for 5'-MD activity by incubation with 0.6 nM [125I]rT3 for 30 min at 37 C in the presence of 10 mM dithiothreitol. The peak 5'-MD activity was detected in fraction 6 (Rf, 0.375). This fraction accounted for about 30% of total 5'-MD activity and only 6% of protein in the gel. The gel slice containing the peak 5'-MD activity was next subjected to a second electrophoresis at pH 7.6 for 4 h in a 90 degrees different direction. The most 5'-MD activity was demonstrated in the third of 8 gel fractions. SDS-polyacrylamide gel electrophoresis of this fraction demonstrated only one 55 K protein in most experiments and occasionally an additional 35 K protein. The specific activity of 5'-MD in this fraction of greater than 11 pmol I-/mg protein.h was at least 2.3-fold that in SRLMP. Rabbits were immunized with agarose gel containing the peak 5'-MD activity. Immunized rabbit immunoglobulin G bound up to 70% of 5'-MD activity in SRLMP, and the binding effect varied in a dose-dependent manner. Western blot analysis revealed that the anti-5'-MD antibody bound only one 55 K protein among innumerable proteins in SRLMP. A specific antiprotein disulfide isomerase antibody bound a slightly larger, 57 K protein in SRLMP.(ABSTRACT TRUNCATED AT 400 WORDS)

Acrylamide↗

The localization and some properties of the N-deacetylase of Ascaris lumbricoides var suum.

1. Hydrolysis of acetanilide by the nematode Ascaris lumbricoides var suum occurred in the cytosol of the intestinal epithelium and to a lesser degree in the cytosol of the reproductive organs. Cuticle and mesenchyme fluid showed no measurable activity. 2. The molecular weight of the enzyme responsible for hydrolysis was estimated to be about 90 000. 3. Hydrolysis of N-acetyl compounds occurred optimally at pH 7-4 at 30-35 degrees and at a substrate concentration of 10(-2) M. 4. The reaction was inhibited 50% by Cu2+, Zn2+, n-ethylmaleimide or p-chloromercuribenzoate at 10(-4) M. The anthelmintic organophosphates inhibited N-deacetylation by 50% at 10(-7) M. Dithiothreitol enhanced the reaction rate but glutathione and thioglycollic acid were without effect.

Acetanilides↗

Gold alloys for resin bonding including small amount base metals--structural changes of alloy surface by the high-temperature oxidation.

To achieve durable bonding with adhesive resin, the surface roughness and the kinds of oxides, respectively to increase mechanical retention to enhance the chemical affinity of adhesive monomer with the gold alloy, were regulated by high-temperature oxidation together with the addition of small amounts of base metals. Alloys containing 2 mass% of Ni, In, or Cr with Cu were oxidized at 800 degrees C for 20 min in air, pickled in thioglycolic acid, and subsequently oxidized at 500 degrees C for 10 min in air. The morphology of the internal oxidation zone changed markedly according to the added base metals. Although the internal oxide particle composed of only Cu2O was removed by pickling, NiO, In2O3, and chromium oxides could not removed and remained on the alloy surface. The surface roughness was increased by addition of Ni, In, or Cr. Applying the present method can control the roughness and chemical states on a gold alloy surface to increase its adhesive ability with adhesive resins.

Acrylic Resins↗

Development of a new medium for the isolation of Arcobacter spp.

Arcobacter, the newly reclassified Campylobacter species, has been shown to cause diarrhea in both humans and animals. Few studies have been conducted regarding its occurrence in foods because of the lack of effective isolation and identification methods. The purpose of this study was to develop a plating medium that would be selective for the three most commonly found Arcobacter species. The effect of common components used in media intended for the isolation of Campylobacter, Helicobacter, and other gram-negative rods was examined. These components were divided into five distinct groups: (1) basic growth nutrients, (2) reducing and growth-promoting agents, (3) detoxifying agents, (4) antibiotics, and (5) color-enhancing compounds. Components from each of these groups were tested for their ability to recover Arcobacter on a solid medium when incubated aerobically at 30 degrees C for up to 72 h. Growth was evaluated by the ecometric technique, colony size, and differential colony morphology after incubation. After initial evaluations, five formulas showing the best results were selected and tested in detail and compared with brucella agar. A medium containing a basal nutrient mix along with 0.05% thioglycolic acid, 0.05% sodium pyruvate, and 5% sheep's blood (pH 6.9+/-0.2) was found to be the most effective for the growth of A. butzleri, A. cryaerophilus, and A. nitrofigilis. In addition to superior growth characteristics, a deep red color around the colonies also was observed with this formulation.

Aerobiosis↗

New pleuromutilin derivatives with enhanced antimicrobial activity.II.Structure-activity correlations.

Structural modification of the antibiotic pleuromutilin has afforded several derivatives with considerably enhanced activity against bacteria and mycoplasmas, and has permitted conclusions to be reached about structure-activity relationships. The carbonyl group in the five-membered ring and the hydroxyl group at C11 seem to be essential for activity. The vinyl group can be hydrogenated without loss of activity. Chemical modification at C14 offers the most possibilities for achieving the best activity and solubility properties. Mutilin, and other compounds with a free OH at C14, are inactive. It was shown that mutilin esters of substituted thioglycolic acids had distinctly superior MIC values, especially in combination with a tertiary amino group in the side chain, the latter group of derivatives having MIC values better than pleuromutilin by a factor of more than 10. Further variation within this group led to the development of 14-deoxy-14-[(2-diethylaminoethyl) thioacetoxy]-mutilin hydrogen fumarate (81.723 hfu, tiamulin) for extensive investigation of its chemotherapeutic potential.

Anti-Bacterial Agents↗

Synthesis and reactions of some new substituted 6-imidazolyl-4-oxo-4H-1-benzopyran-3-carboxaldehyde and use of DNA in evaluation of their biological activity.

Vilsmeier-Haack reaction of imidazolyl acetophenone I gave 6-imidazolyl-4-oxo-4H-1-benzopyran-3-carboxaldehyde II. The compound II was reacted with primary amines (1:1 molar ratio) to form the corresponding n-aril (meteroaryl) imino derivatives IIIa-f. Treatment of aldehyde II with excess amines (1:2 molar ratio) gave the corresponding 2-arylamino-3-arylaminomethylenebenzopyran derivatives IVa-c. The n-aril (meteroaryl) imino derivatives IIIb,d,e,f were reacted with thioglycollic acid to give benzopyranothiazepinone derivatives VIa-d. When the aldehyde II was treated with secondary amines gave the corresponding trans-enaminoketones VIIIa-c. Trans-enaminoketones VIIIa-c were reacted with hydrazines and/or hydroxylamine hydrochloride to give pyrazolyl and/or isoxazolyl benzene IXa-c and X, respectively. The reaction of aldehyde II with hydrazines on cold gave the corresponding hydrazones XIIIa-d. However, the reaction of aldehyde II with hydrazines on refluxing gave the corresponding pyrazole derivatives 5 XIVa,b and XVa,b.. The structural formula of the new compounds were established by using different instrumental analyses. Some compounds in this study were biologically evaluated for their ability to bind to DNA.

Benzopyrans↗

Synthesis of some thiazolyl and thiadiazolyl derivatives of 4(3H)-quinazolinone as anti-inflammatory-antimicrobial agents.

This paper describes the synthesis of four series of 4(3H)-quinazolinone derivatives. The first series was prepared by cyclization of the intermediate 3-aryl-2-substituted thiocarbamoylhydrazonomethyl-4(3H)-quinazolinones 2a,b with ethyl bromoacetate to afford the corresponding thiazolidinonyl derivatives 3a,b. The second series were prepared by the cyclization of the intermediate 2a,b with phenacyl bromide or 4-substituted phenacyl bromide giving rise to thiazolinyl derivatives 4a-f. Furthermore, the thiazolidinonyl derivatives 5a,b were obtained by reaction of the intermediate 2a,b with thioglycolic acid. On the other hand, heating the intermediate 2a,b with acetic anhydride afforded the corresponding thiadiazolinyl derivatives 6a,b. Some of the synthesized compounds showed promising anti-inflammatory-antimicrobial activities.

Animals↗

Tetrazolo[1,5-a] quinoline derivatives as anti-inflammatory and antimicrobial agents [1].

Tetrazolo[1,5-a]quinoline derivatives bearing in the 4-position various thiazolidinone 3a-c, 5a-c and 7a-c, thiazinone 8a,b, thiazoline 9a-d and thiadiazoline 10a,b moieties have been synthesized and evaluated for anti-inflammatory activity and antimicrobial properties. The synthetic routes involved the reaction of tetrazolo[1,5-a]quinoline-4-carboxaldehyde 1 with amines and hydrazines to give the corresponding aniles 2a-c and hydrazones 4a-c respectively The latter compounds when treated with thioglycolic acid, furnished the corresponding thiazolidinone derivatives 3a-c and 5a-c. Moreover; thiosemicarbazone 6a-c derivatives were subsequently cyclized by various reagents giving rise the title compounds. Some of the products proved to possess potent anti-inflammatory and antimicrobial properties.

Animals↗

Contact dermatitis in hairdressers, 10 years later: patch-test results in 300 hairdressers (1994 to 2003) and comparison with previous study.

BACKGROUND: In the last 20 years, the hairdressing profession has undergone important modifications, mainly because of a change in the substances and techniques used and improved occupational education. OBJECTIVE: To evaluate the modifications in the hairdressing profession and its actual risk of occupational allergic contact dermatitis (OACD). METHODS: We studied all 300 hairdressers seen in our department from 1994 to 2003 and compared the results with those of a previous study of 379 hairdressers who attended our department from 1980 to 1993. All were patch-tested with the European Standard series and specific hairdressing products. As previously, most of the workers were women (93%), with a mean age (23.7 years) slightly higher than that of the workers in our previous study. RESULTS: We found a significant increase in the frequency of positive patch-test responses (78.3% vs 58.8%) and OACD (58% vs 48.8%) with respect to our previous study. We also observed a significant increase in sensitization to most allergens, including p-phenylenediamine base (54% vs 45.9%), 4-aminobenzene (40.7% vs 31.9%), ammonium thioglycolate (2.7% to 12.3%), ammonium persulfate (7.9% to 14.3%), p-toluenediamine sulfate (6.8% to 15.3%), p-aminodiphenylamine (2.9% to 7.7%), o-nitro-4-phenylenediamine (2.1% to 7.3%), and aminophenols (0% to 9%), whereas a decrease was found in sensitization to Disperse Orange (17% vs 32.7%) and thioglycolic acid (15.3% to 3%). CONCLUSION: The high frequency and increase of sensitizations among hairdressers require urgent measures to improve protective measures and their application.

Adolescent↗

[Protected mercaptoalkylpyrimidinones: synthesis and test for immunostimulating activity].

Protected mercaptoalkylpyrimidinones: synthesis and test for immunostimulating activity 3-Hydroxyalkyl-pyrimidine 1 reacts with phosphoroxychloride and thioglycolic acid or thiourea to yield pyrimidin-3-ylalkylthioacetic acids 3 or pyrimidin-3-ylalkylthiouroniumsalts 5 respectively. Some of the pyrimidines 3 and 5 showed immunomodulatory activity.

Adjuvants, Immunologic↗

[The antigenic structure of ferritin. 1. Isolation of ferritin subunits and their immunochemical characteristics].

A method for the isolation of monomers of ferritin subunits has been developed. The procedure comprises dissociation of ferritin by treatment with thioglycolic acid in the presence of phosphate ions and subsequent gel-permeation chromatography. Ferritin and a number of its structural analogues (apoferritin, carboxymethylated ferritin, H- and L-subunits of ferritin) have been immunochemically characterized. The immunoreactivity of ferritin is shown to vary along with the degree of denaturation. Isolation of monomers of H- and L-subunits results in appearance of new antigenic sites. These "hidden" antigenic determinants are presumed to be localized in the regions of intersubunit contacts and intracapsular surface of the ferritin molecule and are responsible for the differences in immunochemical properties of its H- and L-subunits.

Apoferritins↗

Synthesis of some N-substituted indole derivatives and their biological activities.

Acylation of 2,3-diphenyl-5-methoxy-indole using ethyl chloroformate or chloroacetyl chloride in dimethylformamide and sodium hydride yielded the N-substituted derivatives 1 and 2, respectively. While Friedel-Crafts acylation using chloroacetyl chloride afforded di-4,6-chloroacetyl derivative 3, the reaction of the N-chloroacetyl derivative 2 with amines, hydrazines, urea, semicarbazide hydrochloride, thiophenol, benzimidazole-2-thiol, thiosemicarbazide, 2-mercaptoethanol and thioglycolic acid was studied. Several of the compounds were tested for their effect on arterial blood pressure, antiinflammatory and ulcerogenic activities.

Animals↗

Synthesis and biological activity of some new 5-hydrothiazolo[4,3-b]-1,3,4-oxa(thia)diazoles and 5-hydrothiazolo[3,4-b]-1,2,4-triazoles containing 1,2,3-selena(thia)diazole moiety.

p-Acetylphenyl thiosemicarbazide (I) was reacted with different aldehydes yielding the corresponding semicarbazones II. Addition condensation of thioglycolic acid with compounds II gave 4-thiazolidinones III. Reaction of compounds III with conc. H2SO4, KI/I2-NaOH and NaOH afforded the cyclized products (IV-VI). Compounds (IV-VI) reacted with semicarbazide to give the corresponding semicarbazones (VII-IX) which either were oxidized with selenium dioxide or thionyl chloride to give 1,2,3-selenadiazoles (X-XII) and 1,2,3-thiadiazoles (XIII-XV) respectively. Some of the prepared compounds were tested against some strains of bacteria.

Chemistry, Pharmaceutical↗