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A toluidine blue-membrane filter method for the quantitative staining of bacteria.

A method for measuring the uptake of toluidine blue by bacteria on membrane filters was developed. Bacteria were filtered out of solution onto a cellulose acetate filter and stained on the filter at 50 C with toluidine blue in citrate-phosphate buffer, pH 4.0. The filter was destained in ethanol, placed on a glass slide and subsequently made transparent in a 1,4-dioxan and cyclohexanone mixture. The absorbance of the stained bacteria on the slide was measured in a spectrophotometer at 590 nm. The uptake of dye by cells of Streptococcus cremoris and Escherichia coli could be explained using the Freundlich adsorption isotherm. Cell concentrations of both these organisms can be determined with this technique.

Bacteriological Techniques↗

Influence of ketamine, phenylcyclidine, and phenobarbital on cholesterol metabolism in rats.

The effects of ip injections of phenobarbital (100 mg/kg), phenylcyclidine (Sernylan; [1-(1-phenylcyclohexyl)-piperidine-HBl] (1 mg/kg), and ketamine (Ketaset; [dl)2-O-chlorophenyl)-2-(methylamino)cyclohexanone-HCl] (1 mg/kg) on lipid metabolism in rats were compared. This study was undertaken to determine whether the two sedatives currently used in primates share any of the undesirable effects of phenobarbital on lipid metabolism. All three compounds were administered to male Wistar rats for 6 days. Phenobarbital was hepatomegalic, stimulated 7alpha hydroxylation of cholesterol, and inhibited cholesterol synthesis by rat liver slices from mevalonate, but not acetate. The two other sedatives exhibited effects very similar to those observed in the controls. From our work in rats it is concluded that the use of Sernylan or Ketaset for sedation of nonhuman primates will not significantly affect these parameters of lipid metabolism.

Animals↗

Synthesis of trifluoromethylisoxazoloazines.

When reacted with mixtures of phosphorus pentoxide, aniline, and triethylamine hydrochloride, 5-acetamidoisoxazoles (2 and 3) gave isoxazolo[5,4-d]pyrimidines (4 and 7, respectively). The same reagent and cyclohexanone were used to prepare the isoxazoloquinoline 8.

Amines↗

Epiderstatin, a new inhibitor of the mitogenic activity induced by epidermal growth factor. II. Structure elucidation.

The structure of a novel antibiotic, epiderstatin, was determined as 4-[3-((Z)-3,5-dimethyl-2-oxopiperidin-6-ylidene)-2-oxopropyl]-2, 6-piperidinedione by spectroscopic analyses of 1H NMR, 13C NMR, 1H-1H correlation spectroscopy (COSY), 13C-1H COSY, heteronuclear multiple bond correlation spectroscopy, UV and IR spectra. The antibiotic belongs to the glutarimide antibiotics, however, the characteristic feature of epiderstatin is that it has a piperidone ring instead of a cyclohexanone ring.

Antibiotics, Antineoplastic↗

Biomolecular-chemical screening: a novel screening approach for the discovery of biologically active secondary metabolites. III. New DNA-binding metabolites.

Based on the chemical screening technique, biomolecular-chemical screening has been developed which makes use of two-dimensional TLC analysis of microbial extracts and combines thin-layer chromatography (RP-18) with binding studies towards DNA. In the first dimension the metabolites of the crude microbial extract are separated, and in the second dimension binding properties towards DNA are analysed. An initial screening program with 500 microbial extracts prepared by solid-phase extraction with XAD-16 resin resulted in 17 samples which contained metabolites with significant DNA-binding behavior. Fermentation, isolation and structural characterization led to already known metabolites [phenazine-1,6-dicarboxylate (1), phencomycin (2), 11-carboxy-menoxymycin B (3), soyasaponine I (4), and (8S)-3-(2-hydroxypropyl)-cyclohexanone (5)], as well as to new secondary metabolites. Fermentation of the producing organisms of the new DNA-binding metabolites, ent-8,8adihydro-ramulosin (6). (2R,4R)-4-hydroxy-2-(1,3-pentadienyl)-piperidine (7), (5R)-dihydro-5-pentyl-4'-methyl-4'-hydroxy-2(3H)-furanone (8), and seco-4,23-hydroxyoleane-12-en-22-one-3-carboxylic acid (9), as well as isolation, structural characterization, and physico-chemical properties are reported.

Chromatography, Thin Layer↗

[The effect of chemical noxae on the health status of a group of workers in the chemical industry of Săvineşti].

The paper presents the study on the effect of the industrial xenobiotics against some biochemical indicators, in view to detect, precociously, the probable significant changes. There are presented the medium, minimum, maximum and median value for the following parameters: total proteins, haemoglobin and glucose for 112 workers exposed or not exposed to specific noxae--benzen, phenol, cyclohexanone. The analysis of the results shows decreased values for some workers, even at the inferior limit, in rapport with the normal values, for the parameters studied. These persons have been directed for complex investigations, in view to determine the causes and to establish the adequate treatment.

Blood Glucose↗

Flow injection spectrophotometric determination of copper, iron, manganese, and zinc in animal feeds using a common manifold.

A rapid and simple procedure was developed for the determination of copper, iron, manganese, and zinc in animal feeds using an identical flow injection spectrophotometric manifold but different chromogenic reagents and different detection wavelengths. Bis(cyclohexanone)oxalydihydrazone, formaldoxime, 1,10-phenanthroline, and xylenol orange were adopted as chromogenic reagents for Cu, Mn, Fe, and Zn, respectively. Detection conditions such as manifold parameters, buffer pH, reagent concentration, temperature, and acidity of sample solution were optimized. Analytical characteristics of the method and interference of metal ions commonly present in feeds were studied. By changing the reagents and detection wavelengths, which can be done quickly, the proposed low cost flow injection system can determine Cu, Fe, Mn, or Zn in the range of 0.5-10 mg/L with a sampling throughput of 120/h.

Animal Feed↗

Enthalpy changes observed upon mixing liquid (R)- and (S)-enantiomers at 298.15 K.

Enthalpies of mixing of (R)- and (S)-enantomers of liquid chiral compounds such as 5-isopropenyl-2-methyl- 2-cyclohexanone (1), glycerol acetonide (2), 4-isopropenyl-1-methylcyclohexene(3), 2,6,6-trimethyl-bicyclo[3.1.1] hept-2-ene (4) have been measured over the whole range of mole fractions at 298.15 K, albeit very small values. Mixing of R-1 and S-1 realises enthalpic stabilisation over the whole range of mole fractions, whereas that of R-2 and S-2, R-3 and S-3, or R-4 and S-4 realise enthalpic destabilisation over entire compositions. The maximum values of enthalpies of mixing and the intermolecular interaction of bond dipoles obtained by the molecular mechanics calculations showed a linear correlation, except the compound having two chiral centers or a phenyl group.

Journal Article↗

Identical flow injection spectrophotometric manifold for determination of protein, phosphorus, calcium, chloride, copper, manganese, iron, and zinc in feeds or premixes.

A simple procedure using an identical manifold was developed for determination of nitrogen (protein) phosphorus, calcium, chloride, copper, manganese, iron, and zinc in feeds and feedstuffs. By changing appropriate reagents and detection wavelength, these 8 elements were determined successively with a simple identical double-line flow injection (FI) manifold. Fl spectrophotometric determinations were made by the blue indophenol reaction for ammonium, the molybdenum blue method for phosphate, the cresolphthalein complexone procedure for calcium, and the mercuric thiocyanate procedure for chloride. The chromogenic reagents for copper, iron, manganese, and zinc determination were bis(cyclohexanone)oxalydihydrazone (Cuprizone), 1,10-phenanthroline, formaldoxime, and xylenol orange, respectively. Sample digestion catalyst, Fl manifold, and some chemical parameters were optimized. The proposed procedure had a sampling rate of 90/h for each analyte. The determination ranges (mg/L) were 10-60 for N, 1-15 for P and Ca, 540 for Cl, and 0.5-15 for Cu, Fe, Mn, and Zn, respectively. Results of the analyses of animal feed and feedstuff samples by this procedure did not differ significantly from those obtained by proven manual methods.

Animal Feed↗

Determination of amphetamine by Schiff base formation and quantitative gas-liquid chromatography.

A procedure was developed to determine amphetamine salts in solid dosage forms. Amphetamine was extracted from the solid matrix with dilute hydrochloric acid and reacted with cyclohexanone in a strongly basic aqueous methanolic solution. The Schiff base reaction product was extracted with hexane for gas chromatographic determination. Reaction time and optimum conditions were studied. Phenethylamine, similarly treated, was used as an internal standard. Results compared favorably with those obtained by using USP methods.

Amphetamine↗

[Ketamine--dreams and realities].

Ketamine is an anaesthetic used in human medicine and veterinary practice, synthesised on 1962 and marketed on 1970 in France. Recreational uses were described during 1992 in the medical community and in 1996 in the dance settings. The chemical name of ketamine is 2--(2chlorophenyl)2-(methylamine)-cyclohexanone, an aryl cyclohexylamine, structurally related to phencyclidine. Ketamine is known under the following street names: Keta K, Kate, Special K, Vitamin K, la Golden, la Vétérinaire. Ketamine is used intranasally, orally and intramusculary in recreational use. Ketamine is manufactured by the chemical industry. Due to the complicated synthesis, it is sold illicitly for recreational use. Ketamine is a dissociative drug, and the user enters in a psychedelic dream with hallucinations, floating sensation, feeling of dissociation of the mind from the body. The dream is forgotten, the user full in reality with loss of self control, risk of acute intoxication. In long-term exposure, tolerance, dependence, withdrawal signs and flash back are described. Ketamine trademarks are subject to control in France through medicine legislation Ketamine and its salts are subject to control under the national legislation on narcotics and psychotropics substance. From September 2001, the theft of medical and veterinary trademarks have to be declared to police, care health authority Pharmacy control authority and French Health Products Safety Agency.

Anesthetics, Dissociative↗

[Determination of 1,1'-peroxydicyclohexylamine by gas chromatography].

A method for the determination of 1,1'-peroxydicyclohexylamine and cyclohexanone by gas chromatography was investigated. The conditions were optimized by changing the stationary liquid, internal standard and injector temperature. The gas chromatographic conditions were as follows: flame ionization detector; 10% OV-101 column; the injector temperature, 140 degrees C; temperature programming; internal standard, dodecane. All components and internal standard were separated in 11 min. The method is simple, rapid, sensitive and reproducible.

English Abstract↗

Syntheses and biological activities of (+/-)-streptovitacin A and E-73.

(+/-)-Streptovitacin A (1) and its stereoisomers were synthesized by an aldol reaction of (+/-)-2,4-dimethyl-4-trimethylsiloxy-1-cyclohexanones (4b and 9) with 4-(2-oxoethyl)-2,6-piperidinedione (5). E-72 (2) was derived from synthetic 1. (+/-)-1 showed moderate growth inhibition against fungi and lettuce seeds.

Antifungal Agents↗

Synthesis and antidepressant activity of some new 8-thiocarbamoyl-7,8-diazabicyclo[4.3.0]non-6-ene derivatives.

Several 8-thiocarbamoyl-7,8-diazabicyclo[4.3.0]non-6-enes were prepared by condensing two alpha,beta-unsaturated ketones (2-benzylidene and 2,6-dibenzylidene cyclohexanones) with hydrazine hydrate and reacting the resulting compounds with various isothiocyanates. The structures of the synthesized compounds were confirmed by spectra (IR, 1H-NMR) and microanalysis; they were tested for antidepressant activity using Porsolt's behavioural despair test.

Animals↗

Analysis of occupational risk due to exposure to carcinogenic factors in the work environment of a chemical plant.

The evaluation of exposure to some carcinogenic chemicals and seemingly carcinogenic chemicals, e.g. mist of sulphuric acid, benzene, chromium (VI) and formaldehyde among employees of some departments of a chemical plant was carried out. The study was carried out in the years 1999--2002. The analysis of concentration of the studied substances revealed the highest risk of exposure in the Cyclohexanon Department and The Maintenance Services Department in Caprolactam department. No direct relationship between the exposure to the studied chemicals and the development of cancer in the working population was found.

Carcinogens, Environmental↗

Experimental demyelination caused by primary oligodendrocyte dystrophy. Regional distribution of the lesions in the nervous system of mice [corrected].

BACKGROUND AND PURPOSE: Heterogeneity of multiple sclerosis lesions has been recently indicated: In addition to T-cell-mediated or T-cell plus antibody-mediated autoimmune mechanisms (patterns I-II) two other patterns (IIl-IV) were described. Patterns Ill-IV are characterized by primary oligodendrocyte dystrophy, reminiscent of virus- or toxin-induced demyelination rather than autoimmunity. It was described more than 30 years ago that dietary application of a copper-chelating agent called cuprizone results in primary oligodendrocyte degeneration which is followed by demyelination. The aim of the present study was to examine the regional distribution of cuprizone induced oligodendrocyte dystrophy and demyelination in the nervous system of mice. MATERIAL A METHODS: Demyelination was induced in male weanling Swiss-Webster mice by feeding them on a diet containing 0.6% (W/W) cuprizone bis(cyclohexanone)-oxalyldihydrazone (G. F. Smith Chemical, Columbus OH) for 8 weeks. Animals were sacrificed after 3, 7, 14, 27, 35, 56 days of cuprizone administration. Samples were taken from corpus callosum, anterior commissure, optic nerve, cervical spinal cord and sciatic nerve. Samples were examined by immunohistochemistry, in situ hybridization for myelin proteins and myelin protein mRNA-s, respectively. Conventional neuropathological stainings and electron microscopy was also performed. RESULTS: Oligodendrocyte degeneration and demyelination followed a particular standard pattern in the central nervous system. Profound myelin loss developed in the superior cerebellar peduncle, anterior commissure and corpus callosum, whereas the optic nerves, velum medullare anterior and spinal cord showed little or no demyelination. Sciatic nerves were unaffected. No infiltration by lymphocytes or blood-brain barrier damage was observed during cuprizone treatment. CONCLUSION: Cuprizone induced oligodendrocyte damage and demyelination follows a particular standard pattern in the central nervous system of mice. Cuprizone induced demyelination might be considered as a model for human demyelinating disorders with primary oligodendrocyte dystrophy and apoptosis.

Animals↗

Toxicity potential of compounds found in parenteral solutions with rubber stoppers.

Leached stopper components found in parenteral solutions produced by several manufacturers were identified and quantitated. Their toxicity potential was determined by comparing the types and quantities of the leached components with known toxicity levels and/or harmful effects. Toxicity potentials for benzaldehyde, 2-butoxyethanol, cyclohexanone, ethylbenzene, 1,1,2,2-tetrachloroethane, and tetrachloroethylene are listed. Breakdown products of dextrose (furfural and 5-hydroxymethylfurfural), which may also have harmful effects, were quantitated.

Drug Contamination↗

[Pollution survey of carbonyl compounds in train air].

The methods for sampling and analyzing 10 carbonyl compounds such as formaldehyde, acetaldehyde, acrolein, acetone, propionaldehyde, crotonaldehyde, butyraldehyde, benzaldehyde, cyclohexanone, valeraldehyde in indoor air were proposed with the sampling efficiency, recovery and detection limit being 92% - 100%, 91% - 104%, 0.26ng/m3 - 0.82ng/m3, respectively. Pollution survey of 10 carbonyl compounds in air of 6 trains was conducted. It was indicated that the total concentrations of carbonyl compounds were 0. 1591 mg/m3 - 0.2828 mg/m3 with the average concentration of 0.2330 mg/m3, the average concentrations of formaldehyde, acetaldehyde and acetone were 0.0922 mg/m3, 0.0499 mg/m3 and 0.0580 mg/m3, accounted for 42.6%, 21.4%, 24.9% of the total carbonyl compounds' concentrations, respectively. The carbonyl compounds probably came from woodwork and tobacco. The intake of carbonyl compounds for the passengers was approximately 0.043 mg/h - 0.076 mg/h.

Acetaldehyde↗