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Biomedical subjects

L Weber

Publications and source records attributed to L Weber.

At least 19 recordsLinked to original sources

Determination of benzene metabolites in urine of mice by solid-phase extraction and high-performance liquid chromatography.

A method was developed for quantitative measurement of trans,trans-muconic acid, catechol, hydroquinone and phenol in urine. Hydrolysis of esterified and glucuronized phenolic compounds was effected by specific enzymes. The hydrolysed mixture was purified and separated by solid-phase extraction with an anion exchanger, followed by extraction with diethyl ether. By using a clean-up procedure the natural background from mouse urine could be reduced, so that the detection limit of the metabolites was in the range 3-60 mg/l. Optimization of the chromatographic conditions resulted in a short high-performance liquid chromatography analysis time. Phenol had the longest retention time of about 10 min. The clean-up procedure could also be used for phenylmercapturic acid, an additional benzene metabolite, but for sensitive high-performance liquid chromatographic detection of phenylmercapturic acid other conditions are necessary.

Animals

Gas chromatographic determination of urinary phenol conjugates after acid hydrolysis/extractive acetylation.

Phenolic metabolites of inhaled aromatic solvent vapours were liberated by acid hydrolysis of their urinary conjugates. Steam distillation enhanced by salting-out with MgSO4 gave good recoveries. After extractive acetylation, the derivatives of all cresols and xylenols were completely separated on a Se-54 capillary column. The overall recoveries of urinary phenols relative to the internal standard, 3-chlorophenol, were in the range 92-99%.

Acetylation

Hematological toxicity of tetrachloroethylene in mice.

Female C57/BL/6 x DBA/2 hybrid mice were exposed to two concentrations (270 and 135 ppm) of tetrachloroethylene (PER) in inhalation chambers for 6 h/day, 5 days per week, up to 11.5 weeks (270 ppm) and 7.5 weeks (135 ppm), respectively, followed by a 3-week exposure-free period. In the peripheral blood a reduction of lymphocytes/monocytes and of neutrophils was observed with an almost complete regeneration in the exposure-free period. A reticulocytosis during and also after the exposure indicated a compensatory reaction in the erythroid cell system. Bone marrow pluripotent stem cells (CFU-S) were not affected up to 7.5 weeks. Erythroid committed cells (BFU-E and CFU-E) did react, with a reduction of CFU-E numbers at 7.5 and 11.5 weeks. A slight reduction of CFU-C numbers at 7.5 weeks indicated, in accordance with the peripheral blood data, a disturbance of the granulocytic cell series as well.

Animals

A RAD9-dependent checkpoint blocks meiosis of cdc13 yeast cells.

Mutations in CDC13 have previously been found to cause cell cycle arrest of Saccharomyces cerevisiae at a stage in G2 immediately preceding the mitotic division. We show here that cdc13 blocks the meiotic pathway at a stage that follows DNA replication, but in this case the spindle has not yet formed nor have the chromosomes undergone synapsis or recombination. This arrest is alleviated by rad9, thus implicating the same checkpoint function that delays mitotic progression when chromosomal lesions are present. An assessment of the spores produced upon alleviation of the meiotic arrest by rad9 reveals that the absence of recombination in strains bearing cdc13 alone is attributable to the RAD9-mediated arrest rather than to other effects of cdc13 lesions. We have tested the possibility that this checkpoint function is important in regulating meiotic progression to permit resolution of recombinational intermediates during ongoing meiosis and have found no evidence that rad9 alters the execution of functions that might depend upon such regulation. We consider the possible role of other checkpoints in yeast meiosis.

Cell Cycle Proteins

[Experimental studies on the recovery of anesthetic gases].

The volatile anesthetic agents halothane, enflurane, and isoflurane are chlorofluorocarbons (CFC) and contribute to ozone depletion. Although the contribution is small, its importance is rising, as technical CFCs will be phased out according to the Montreal protocol (1987) and the London conference (1990) by the year 2000. Alternative procedures and CFC-free volatile agents such as des- and sevoflurane do not contribute to depletion of the ozone layer, but will not replace standard methods using volatile anesthetic agents in the near future. METHODS. In an experimental setup, we filtered anesthetic waste gases from scavenging systems of rebreathing circles by activated carbon filters. The filtered substances were desorbed by a heat chamber and condensed in a cold trap. RESULTS. By this method, it was possible to retrieve 50%-60% of the applied gases. Gas chromatographic analysis showed halothane containing traces of pollutants and isoflurane and enflurane as pure substances. DISCUSSION. The retrieval of anesthetic waste gases is easy; no sophisticated technical equipment is necessary. Purity of substances could make recycling possible and offer a method to avoid environmental pollution by volatile anesthetics.

Anesthetics

Cell surface glycoproteins define distinct states of eccrine gland differentiation.

The capability to distinguish eccrine gland cells of the ductal compartment from the secretory portion on the molecular level, as well as from epidermal keratinocytes and other skin cells, is of importance for the study of eccrine differentiation and function. Furthermore, the assessment of differences between the cell systems is useful for the characterization of benign and malignant neoplasms derived from eccrine glands. In the present study, we analysed the expression of four selected epithelial cell surface glycoproteins (gp 80, gp 38, gp 115 and gp 200) in eccrine glands using specific monoclonal antibodies. We found that the glycoproteins are differentially expressed in the ductal compartments and in the secretory portion of the glands, as well as in normal epidermis and other skin cells. This permitted the assignment of specific phenotypes to cells of the ductal compartment and to those of the secretory portion.

Antibodies, Monoclonal

Action of benzene metabolites on murine hematopoietic colony-forming cells in vitro.

Five benzene metabolites--phenol, catechol, hydroquinone, parabenzoquinone, and trans,trans-muconaldehyde--were tested in vitro in murine hematopoietic cell cultures for erythrocytic (BFU-E and CFU-E) and granulocytic (CFU-C) colony growth. The dose range was 4 x 10(-4) to 4 x 10(-8) M. Of these compounds, phenol showed the lowest toxicity. In general, CFU-E cultures were far more sensitive than CFU-C and BFU-E. The results are discussed in relation to the test system used, the in vivo sensitivity of CFU-E, and the lack of knowledge about in vivo concentrations of these compounds.

Animals

Conditional power for arbitrary survival curves to decide whether to extend a clinical trial.

This article describes a computer simulation methodology to calculate conditional power for comparing two arbitrary survival curves as an aid in deciding whether or not to extend a clinical trial. The method is a modification of that by Halpern and Brown, which computes unconditional power for comparing arbitrary survival curves at the beginning of a study. The advantage of this method is that it permits conditional power calculations for comparisons of distribution-free survival curves. Power is computed for two commonly used statistical tests, the log-rank test and Gehan's generalization of the Wilcoxon test, and for a modified Kolmogorov-Smirnov test, which is particularly sensitive to crossing survival curves. This method for estimating conditional power should be useful in the situation in which a decision must be made regarding the benefit of continuing a clinical trial. The application of this method is shown using VA Cooperative Study No. 90, "Prognosis and Outcome Following Heart Valve Replacement."

Aortic Valve

Integrin alpha 2 beta 1 is upregulated in fibroblasts and highly aggressive melanoma cells in three-dimensional collagen lattices and mediates the reorganization of collagen I fibrils.

The ability of cultured human fibroblasts to reorganize and contract three dimensional collagen I gels is regarded as an in vitro model for the reorganization of connective tissue during wound healing. We investigated whether adhesion receptors of the integrin family are involved. It was found that synthesis and transcription of the alpha 2 beta 1 integrin (but not of alpha 1 beta 1 or alpha 3 beta 1) is selectively upregulated when fibroblasts are seeded into type I collagen gels. Time course experiments revealed that high synthetic levels of alpha 2 beta 1 parallel the gel contraction process and return to "baseline" levels after the contraction has subsided. Furthermore, function-blocking mAbs directed to the alpha 2 and beta 1 chain of integrins inhibited gel contraction. Remodelling of connective tissue can be important for tumor cells during invasion and formation of metastases. Therefore, we tested human melanoma cell lines for this function. Five out of nine melanoma lines contracted collagen gels in vitro. Among these, two highly aggressive melanoma cell lines (MV3 and BLM) most efficiently contracted gels almost reaching the rate of normal adult fibroblasts. In these cells, synthesis of alpha 2 beta 1 was also significantly upregulated when seeded into collagen I gels. Moreover, function blocking anti-alpha 2 in conjunction with anti-beta 1 chain mAbs completely inhibited gel contraction for several days. Other melanoma cells (530) with lower metastatic potential which were not able to contract gels, showed no induction of alpha 2 beta 1 synthesis in gel culture. Our results suggest an important role of integrin alpha 2 beta 1 in the contraction of collagen I by normal diploid fibroblasts during wound healing and in the reorganization of collagen matrices by highly aggressive human melanoma cells.

Adult

Identification of a melanoma progression antigen as integrin VLA-2.

The expression of the integrin receptors VLA-1, -2, -3, and -6 was studied in normal cultured melanocytes and in five melanoma cell lines. Normal melanocytes synthesized VLA-3, but did not reveal detectable levels of VLA-1, -2, and -6. All melanoma cell lines, however, expressed VLA-2, -3, and -6. VLA-1 was synthesized by two of five melanoma lines. In parallel, we had analyzed the expression of four previously characterized melanoma cell surface antigens. One of them (antigen A.1.43), which is associated with tumor progression of human melanoma, revealed a striking similarity to VLA-2. In sequential immunoprecipitation experiments, we show that A.1.43 is identical with the integrin VLA-2, a cell surface receptor for collagen, laminin, and fibronectin.

Antibodies, Monoclonal

[Photoallergic reaction to olaquindox].

Olaquindox, an antibiotic belonging to the quinoxaline group, is a growth promoter used in pig breeding. A farmer developed a photoallergic contact eczema to it, which progressed to a persistent light reaction. Besides this, we found a photosensitivity to chlorpromazine, a tranquillizer that is also used in pig breeding. In addition, after using potent light-protective drugs containing UV-A and UV-B filters, this patient developed a (photo)allergy to different filter substances. The photoallergy to olaquindox was caused by mixing a food additive containing olaquindox with the pigswill for both young and adult animals. We point out the dangers (potent photoallergen, cancer induction) resulting from the widespread use of this substance in the EC.

Animal Husbandry

Haemopoietic stem cells in mice chronically exposed to styrene vapour.

Female C57BL/6X DBA/2 hybrid mice were exposed to two concentrations of styrene in inhalation chambers for 6 h/day, 5 days per week. The concentrations were 220 ppm for a period of up to 8 weeks and 440 ppm for 2 days, followed by 330 ppm for up to 8 weeks due to early deaths after 440 ppm. Parallel groups were also treated with 5% ethanol in the drinking water during the days of styrene exposure. In the peripheral blood only a lymphocytopenia was found. The numbers of pluripotent haemopoietic stem cells, CFU-S, were unaffected, as were granuloid committed stem cells, CFU-C. Early and late erythroid stem cell numbers, BFU-E and CFU-E, showed high variability. At 220 ppm about half of the animals had CFU-E numbers below 80% of controls, at 440/330 ppm about two-thirds of the animals had BFU-E and CFU-E numbers below this level. An additional effect of ethanol was not found.

Air

Use of principal components analysis to develop a composite score as a primary outcome variable in a clinical trial. The VA Cooperative Study Group on Cochlear Implantation.

This article describes the use of principal components analysis to derive a composite score from a battery of 24 audiologic tests. The composite score is being used as the primary outcome variable in a clinical trial comparing the efficacy of three cochlear implant devices for people with bilateral, profound hearing loss. The first principal component from the within-class pooled variance-covariance matrix over four time periods was chosen to establish the coefficients for the composite score. This component accounted for 61% of the total variance of the 24 audiologic tests. The first principal component had its largest coefficients associated with the most difficult audiologic tests. The mean composite score of all patients improved over time; some patients showed dramatic improvement. The changes in the composite score over time were also closely related to subjective impressions of implant performance by the patient, audiologist, and otolaryngologist.

Analysis of Variance

Trace analysis of chloramphenicol residues in egg powders by capillary gas chromatography-electron capture detection.

Chloramphenicol (CAP) in egg powder samples was extracted with acetonitrile by sonication. After centrifugation, the supernatant was diluted with water and defatted with n-hexane, and cleanup was performed on a microcolumn of graphitized carbon black (GCB). Different silylation methods were compared for the derivatization of the analytes. Trimethylsilyl N,N-dimethylcarbamate produced stable di-TMS derivatives which were analyzed by capillary gas chromatography-electron capture detection (CGC-ECD) after splitless injection. The meta isomer of the chloramphenicol served as the internal standard for quantitation, and 80% + mean recovery was obtained in the 0.1-10 micrograms/kg (ppb) concentration range.

Acetonitriles