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Biomedical subjects

J Hollender

Publications and source records attributed to J Hollender.

At least 19 recordsLinked to original sources

Contribution to the Detection and Identification of Oxidation Metabolites of Nonylphenol in Sphingomonas sp. strain TTNP3.

Sphingomonas sp. strain TTNP3 has been previously described as a bacterium that is capable of degrading the technical mixture of nonylphenol (NP) isomers and also the 4(3',5'-dimethyl-3'-heptyl)-phenol single isomer of NP. Until recently, 3,5-dimethyl-3-heptanol was the only reported metabolite of 4(3',5'-dimethyl-3'-heptyl)-phenol. A short time ago, the detection of an intracellular metabolite resulting from the oxidation of 4(3',5'-dimethyl-3'-heptyl)-phenol which was identified as 2(3,5-dimethyl-3-heptyl)-benzenediol has been reported. A decisive element for this identification was the occurrence of some slight differences with the two most probable metabolites i.e. 4(3',5'-dimethyl-3'-heptyl)-resorcinol and 4(3',5'-dimethyl-3'-heptyl)-catechol. These facts led us to hypothesise some NIH shift mechanisms explaining the formation of 2(3',5'-dimethyl-3'-heptyl)-benzenediol. In the present work, we describe the steps that led to the detection of these metabolites in the intracellular fraction of Sphingomonas sp. strain TTNP3. The formation of analogous intracellular metabolites resulting from the degradation of the technical mixture of NP is reported. To further elucidate these degradation products, studies were carried out with cells grown with 4(3',5'-dimethyl-3'-heptyl)-phenol as sole carbon source. The description of the syntheses of reference compounds, i.e. 4(3',5'-dimethyl-3'-heptyl)-resorcinol and 4(3',5'-dimethyl-3'-heptyl)-catechol and their comparative analyses with the intermediates of the degradation of 4(3',5'-dimethyl-3'-heptyl)-phenol are presented.

Chromatography, High Pressure Liquid↗

The degradation of alpha-quaternary nonylphenol isomers by Sphingomonas sp. strain TTNP3 involves a type II ipso-substitution mechanism.

The degradation of radiolabeled 4(3',5'-dimethyl-3'-heptyl)-phenol [nonylphenol (NP)] was tested with resting cells of Sphingomonas sp. strain TTNP3. Concomitantly to the degradation of NP, a metabolite identified as hydroquinone transiently accumulated and short-chain organic acids were then produced at the expense of hydroquinone. Two other radiolabeled isomers of NP, 4(2',6'-dimethyl-2'-heptyl)-phenol and 4(3',6'-dimethyl-3'-heptyl)-phenol, were synthesized. In parallel experiments, the 4(2',6'-dimethyl-2'-heptyl)-phenol was degraded more slowly than the other isomers of NP by strain TTNP3, possibly because of effects of the side-chain structure on the kinetics of degradation. Alkylbenzenediol and alkoxyphenol derivatives identified as metabolites during previous studies were synthesized and tested as substrates. The derivatives were not degraded, which indicated that the mineralization of NP does not proceed via alkoxyphenol as the principal intermediate. The results obtained led to the elucidation of the degradation pathway of NP isomers with a quaternary alpha-carbon. The proposed mechanism is a type II ipso substitution, leading to hydroquinone and nonanol as the main metabolites and to the dead-end metabolites alkylbenzenediol or alkoxyphenol, depending on the substitution at the alpha-carbon of the carbocationic intermediate formed.

Biodegradation, Environmental↗

Degradation of a nonylphenol single isomer by Sphingomonas sp. strain TTNP3 leads to a hydroxylation-induced migration product.

Sphingomonas sp. strain TTNP3 degrades 4(3',5'-dimethyl-3'-heptyl)-phenol and unidentified metabolites that were described previously. The chromatographic analyses of the synthesized reference compound and the metabolites led to their identification as 2(3',5'-dimethyl-3'-heptyl)-1,4-benzenediol. This finding indicates that the nonylphenol metabolism of this bacterium involves unconventional degradation pathways where an NIH shift mechanism occurs.

Benzene Derivatives↗

Efficiency of nanofiltration for the elimination of steroids from water.

Steroid hormones such as the synthetic 17-alpha-ethinylestradiol often persist in the sewage treatment process and metabolites of the natural steroids like estrone from estradiol accumulate resulting in relevant amounts in the aquatic environment. Since biological removal of steroids in the sewage treatment plant is not successfully complete, physical processes such as membrane filtration may become more important. We investigated the elimination of natural and synthetic steroids by nanofiltration using a laboratory membrane reactor. Chemical analysis of 17-beta-estradiol, estrone, estriol, 17-alpha-ethinylestradiol, mestranol, diethylstilbestrol, progesterone and beta-sitosterine was performed after solid phase extraction by GC-MS with standard addition. The elimination rate depended on the nanofiltration membrane material. LFC1 membrane consisting of polyamide removed the steroids over 99% whereas PES10 membrane consisting of hydrolysed polyethersulfone was less efficient, obviously caused by different pore sizes and permeability of the membrane structure.

Animals↗

Microbial degradation of a single branched isomer of nonylphenol by Sphingomonas TTNP3.

The endocrine disrupting chemical nonylphenol (NP) is a technical product which consists of a complex mixture of nonylphenols with different alkyl side-chain isomers. Since the bio-degradation of each NP isomer may lead to its own range of metabolites, the isolation and identification of transformation products is very difficult. In order to overcome this difficulty, the nonylphenol isomer 4(3',5'-dimethyl-3'-heptyl)-phenol (p353NP) was synthesized, and its degradation by an axenic culture of Sphingomonas TTNP3 was investigated with [ring-U-14C]-labelled and non-labelled p353NP including a time-course study. Radioactive mass balancing resulted in different polar soluble fractions, in insoluble radioactivity associated with biomass, and volatile radioactivity in the form of the mineralization product 14CO2. In the extracellular media, the presence of nonanol corresponding to the nonyl chain of the NP isomer was confirmed and its concentration was determined during the course of fermentation. No other radioactive compounds were detected beside the parent isomer. Radioactive metabolites were only found in the intracellular fraction of S. TTNP3.

Biodegradation, Environmental↗

Effect of sample preparation on the in vitro genotoxicity of a light curable glass ionomer cement.

The glass ionomer cement Vitrebond showed a clear genotoxic effect in the in vitro Mammalian Cell Gene Mutation Test (HPRT Test) with CHO cells as well as in the bacterial umu-test with Salmonella typhimurium TA1535/pSK1002. Both DMSO and Ham's F12 cell culture medium extracts according to ISO 10993-12 (Biological evaluation of medical devices-Part 12: sample preparation and reference materials, Geneva, Switzerland) exhibit a clear genotoxic effect in the umu-test. The effect is independent of the extraction volume in a range from 0.5 to 4 ml Ham's F12 cell culture medium. Subsequent extractions of Vitrebond showed no significant difference in the genotoxic response although weight loss and content of 2-hydroxyethyl-methacrylate dropped significantly. In vivo conditions of Vitrebond were simulated by extractions with artificial and collected human saliva. These extracts showed a clear genotoxic effect in the umu-test, even if only a few seconds of extraction time were applied. In conclusion, sample preparations for genotoxicity testing according to ISO 10993-12 reflect the in vivo conditions of Vitrebond applications. This seems to be mostly due to the hydrophilic nature of the genotoxic ingredients.

Animals↗

Selective enrichment and characterization of a phosphorus-removing bacterial consortium from activated sludge.

Under alternating aerobic/anaerobic conditions and without additional carbon sources, a bacterial consortium consisting initially of 18 bacterial strains was obtained in a sequence batch reactor. The phosphorus removal capability could only be maintained using sterile filtrate of activated sludge as medium. The addition of calcium and magnesium salts, as well as vitamins and trace elements, to autoclaved sterile filtrate of activated sludge was not sufficient to achieve stable phosphorus removal. A further enrichment by subcultivation on solid, agar, freezing, and shortening of the aerobic and anaerobic phases led to a defined bacterial consortium consisting of four strains. On the basis of physiological and chemotaxonomic characterization, and partial 16S rRNA sequencing, one of the organisms was identified as Delftia acidovorans. A further isolate belonged to the Bacillus cereus group, and the third isolate was identified as Microbacterium sp.. The remaining strain seems to represent a new genus within the Flavobacteriaceae. Under continuous chemostat conditions, this consortium was able to remove up to 9.6 mg P/l phosphate in the aerobic phase and released up to 8.5 mg/l in the anaerobic phase. Up to 25 mg P-polyphosphate/g dry mass was stored under aerobic conditions.

Actinomycetales↗

Microcosms-experiments to assess the potential for natural attenuation of contaminated groundwater.

Groundwater samples from six wells of a former gas plant site were characterised using chemical, microbial and ecotoxicological methods. Degradation studies were performed in batch-culture under aerobic conditions with the groundwater samples containing their autochthonous microflora and original contaminant mixture. The highest O2-consumption (3 mmol 100 ml-1), combined with BTEX (8.3 mg l-1) and naphthalene (171.3 mg l-1) degradation, as well as formation of organic acids was found after N- and P-supplementation with the highest contaminated groundwater sample. The other highly polluted groundwater sample showed no activity obviously because of the toxicity of some compounds. The major part of the PAHs and BTEX was eliminated in the assays with the low contaminated groundwater samples. The results indicate that the microbial degradation capacity and thereby the natural attenuation capacity in each groundwater differ and cannot be assessed simply by chemical, microbial and toxicological data. Additionally activity tests with authentic groundwater samples with and without nutrient supplementation are recommended.

Carbon Dioxide↗

Sensitive indoor air monitoring of formaldehyde and other carbonyl compounds using the 2,4-dinitrophenylhydrazine method.

The toxic potential of formaldehyde and other aliphatic/aromatic carbonyl compounds requires the determination of even low amounts of these compounds in indoor air. The existing DFG-method for workplace monitoring using adsorption at 2,4-dinitrophenylhydrazine (DNPH)-coated sorbents followed by HPLC-UV/DAD analysis of the extract was modified in order to decrease detection limits. The improvement included an increase in volume and rate of the air sampling, testing applicability of different adsorption materials and a decrease of the extraction volume of the hydrazones. 13 DNPH-derivatives could be separated well on a RP18-column followed by UV/DAD-detection at 365 nm. Recovery rates of 70-100% were determined (apart from acetone with 19%) using dynamically produced artifical carbonyl atmospheres. Detection limits of 0.05-0.4 microgram/m3 were reached by this method which are sufficient for indoor air monitoring.

Air Pollutants, Occupational↗

Biomonitoring of environmental polycyclic aromatic hydrocarbon exposure by simultaneous measurement of urinary phenanthrene, pyrene and benzo[a]pyrene hydroxides.

A high-performance liquid chromatographic method with fluorescence detection was developed which enables the simultaneous determination of the urinary polycyclic aromatic hydrocarbon metabolites 3-hydroxyphenanthrene, 1-hydroxypyrene and 3-hydroxybenzo[a]pyrene. The method has small solvent consumption because of the use of a microbore RP C18 column and a relatively short run time. Low detection limits of 0.02 nmol/l for 3-hydroxypyrene to 0.19 nmol/l for 3-hydroxybenzo[a]pyrene were attained. In contrast, the detection limits of alpha-naphthol and 9,10-dihydroxy-9,10-dihydrophenanthrene were not adequate for the determination of environmental exposure. The developed method was successfully used for the analysis of urine samples from children.

Air Pollutants↗

[Airborne molds and their metabolites at workplaces in composting plants].

Airborne fungal contaminants in composting facilities gain more and more importance in regard to health hazards for workers and an emission of spores and microbial metabolites in the environment. Possible health impacts of bioaerosols are widely discussed from both an allergological and a toxicological point of view. The present study investigates in detail the exposure to airborne fungal spores at working places in compost plants. These fungal spores are compared to the spectrum of microbial volatile metabolites with regard to the physiological properties of each fungal species. The results indicate, that the spectrum of microbial volatile organic compounds (MVOC) as well as the production of mycotoxins can be specific for certain species. In addition to the pathogenic and allergological relevance, the fungi may have different toxicological health impacts. However, an evaluation of health effects caused by the great variety of volatile organic compounds remains difficult, since information on their toxicological impact is lacking.

Air Microbiology↗

Degradation of 4-Chlorophenol via the meta Cleavage Pathway by Comamonas testosteroni JH5.

Comamonas testosteroni JH5 used 4-chlorophenol (4-CP) as its sole source of energy and carbon up to a concentration of 1.8 mM, accompanied by the stoichiometric release of chloride. The degradation of 4-CP mixed with the isomeric 2-CP by resting cells led to the accumulation of 3-chlorocatechol (3-CC), which inactivated the catechol 2,3-dioxygenase. As a result, further 4-CP breakdown was inhibited and 4-CC accumulated as a metabolite. In the crude extract of 4-CP-grown cells, catechol 1,2-dioxygenase and muconate cycloisomerase activities were not detected, whereas the activities of catechol 2,3-dioxygenase, 2-hydroxymuconic semialdehyde dehydrogenase, 2-hydroxymuconic semialdehyde hydrolase, and 2-oxopent-4-enoate hydratase were detected. These enzymes of the meta cleavage pathway showed activity with 4-CC and with 5-chloro-2-hydroxymuconic semialdehyde. The activities of the dioxygenase and semialdehyde dehydrogenase were constitutive. Two key metabolites of the meta cleavage pathway, the meta cleavage product (5-chloro-2-hydroxymuconic semialdehyde) and 5-chloro-2-hydroxymuconic acid, were detected. Thus, our previous postulation that C. testosteroni JH5 uses the meta cleavage pathway for the complete mineralization of 4-CP was confirmed.

Journal Article↗

Regulation of chloro- and methylphenol degradation in Comamonas testosteroni JH5.

Comamonas testosteroni JH5 was isolated from a mixed bacterial culture enriched on different chloro- and methylphenols. The strain completely mineralized a mixture consisting of 4-chlorophenol (4-CP) and 4-methylphenol (4-MP). During degradation of the mixture, 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, 4-hydroxybenzoic acid, and 4-chlorocatechol were detected as short-lived intermediates. Mineralization of 4-CP and that of 4-MP occurred successively and were accompanied by diauxic growth, whereas 4-CP and 2-methylphenol were mineralized simultaneously. It was ascertained that neither a reversible enzyme inhibition nor potential toxic intermediates caused the observed diauxie. Some facts support the hypothesis that the successive degradation of 4-CP and 4-MP is regulated on the level of transcription. C. testosteroni JH5 contained a meta-cleaving enzyme when pregrown on 4-CP and the isomeric monomethylphenols. Inactivation of this enzyme in the presence of 3-chlorocatechol was observed.

Biodegradation, Environmental↗

Catalytic center of cyclodextrin glycosyltransferase derived from X-ray structure analysis combined with site-directed mutagenesis.

An X-ray structure analysis of a crystal of mutant Asp229----Ala of cyclodextrin glycosyltransferase from Bacillus circulans (Ec 2.4.1.19) that had been shortly exposed to beta-cyclodextrin showed density corresponding to a maltose bound at the catalytic center. The crystal structure was refined to an R-factor of 18.7% at 2.5-A resolution. The catalytic center is defined by homology with the structurally known alpha-amylases and by the observation that mutants Asp229----Ala and Asp328----Ala are almost inactive. By model building, the density-defined maltose was extended to a full beta-cyclodextrin, which then indicated the general locations of seven subsites for glucosyl units. The catalytically competent residues Asp229, Glu257, and Asp328 are at the reducing end of the density-defined maltose. In the unligated wild-type structure, Glu257 and Asp328 form a 2.6-A hydrogen bond between their carboxylates in an arrangement that resembles those of the catalytically competent carboxylates in acid proteases. Presumably, the first catalytic step is an attack of the proton between Glu257 and Asp328 on the oxygen of the glycosidic bond.

Bacillus↗

Functional recovery from cancer surgery: estimation of expectations.

Level of expected functional recovery from colostomy, laryngectomy and mastectomy was established by the Delphi technique. One hundred fifteen cancer rehabilitation experts responded to 2 rounds of questionnaires and provided their judgments regarding degrees and timing of recovery in 14 functional, physical and psychological areas. Feedback of first round judgments was provided in the second round and 85% agreement was set as the standard for consensus. The mastectomy patient is expected to have the quickest recovery with former level of functioning reached in most areas by 3 months postdischarge. The colostomy patient is expected to be functioning normally in most areas by 6 months postdischarge. The laryngectomy patient is expected to be functioning normally in most areas by 12 months postdischarge. Some functions were judged to have the same rate of recovery for all 3 patient types; for example, the experts agreed that for the 55-year-old patients described in the case abstracts, sexual activities would not reach the preoperative frequency until 12 months postdischarge.

Breast Neoplasms↗