Pyrene degradation by Mycobacterium sp. strain KR2.

A Mycobacterium sp., strain KR2 which was able to utilise pyrene as sole source of carbon and energy was isolated from a polycyclic aromatic hydrocarbon (PAH) contaminated soil originating from the area of a former gaswork plant. The isolate metabolised up to 60% of the pyrene added (0.5 mg/mL) within 8 days at 20 degrees C. Cis-4,5-pyrene dihydrodiol, 4,5-phenanthrene dicarboxylic acid, 1-hydroxy-2-naphthoic acid, 2-carboxybenzaldehyde, phthalic acid, and protocatechuic acid were identified as degradation products. Based on these findings a degradation pathway for pyrene is suggested which is in good accordance with the data published so far on bacterial pyrene metabolism.

Distribution and transmission of Pseudomonas aeruginosa and Burkholderia cepacia in a hospital ward.

Genotyping and antibiotic susceptibility testing were used to analyze Pseudomonas aeruginosa and Burkholderia cepacia strains from sink drain from 14 pediatric patients with cystic fibrosis (CF) and from hospital personnel as part of a 4 week prospective study of strain transmission in a pediatric ward. A total of 87.5% of all washbasin drains were contaminated with P. aeruginosa [10(2) to 10(5) colony forming units (CFU)/ml sink fluid], whereas B. cepacia was found only once in a sink drain. From the eight CF patients already infected with P. aeruginosa upon entering the ward, we isolated six genotypes that were identical with strains found in sink drains of the ward. Four of the 16 members of the personnel had one positive P. aeruginosa hand culture. B. cepacia was never found in patients or on personnel hands. Hand washing in contaminated sinks (> or = 10(3) CFU/ml) led to positive P. aeruginosa or B. cepacia hand cultures. P. aeruginosa or B. cepacia embedded in sputum were transmissable by hand shaking for up to 180 min, whereas both pathogens suspended in physiological saline were transmissable to other hands only up to 30 min. Genotyping of P. aeruginosa revealed strain transmission from CF patients or the environment to other patients or the personnel, as well as one transmission from the environment to a CF patient. The ability of CF sputum to prolong survival of P. aeruginosa and B. cepacia may be important for strain transmission. The results suggest that improved hygienic measures are required to prevent routes of bacterial transmission via the hands and sink drains.

Mitral valve resistance as a hemodynamic indicator in mitral stenosis.

Variability of the valve area calculated by the Gorlin formula has been noted in bioprosthetic and aortic valves, but few data are available for native stenotic mitral valves. Valve resistance has been proposed as an alternative hemodynamic indicator; however, its value in mitral stenosis has not been assessed. Thirty-four patients had simultaneous recordings of left atrial and ventricular pressures, 26 after percutaneous balloon mitral dilatation (PBMD). Patients with shunt or mitral regurgitation were excluded. Mitral valve resistance correlated exponentially with Gorlin mitral area (y = 133*[area]-1.5; p less than 0.0001). Both Gorlin mitral area and mitral resistance improved after PBMD (0.89 +/- 0.07 cm2 to 2.22 +/- 0.15 cm2; p less than 0.001; and 166 +/- 20 to 40 +/- 8 dynes.s.cm-5; p less than 0.001). Gorlin area and mitral resistance correlated with New York Heart Association functional class. After infusion of isoproterenol in 17 patients, there was an increase in Gorlin area (baseline 1.77 +/- 0.22 cm2, change 0.23 +/- 0.10; p less than 0.03), whereas mitral resistance did not change (baseline 96 +/- 16 dynes.s.cm-5, change 2 +/- 5; p = not significant). Mitral resistance is valuable in the assessment of mitral stenosis. It varies less than Gorlin mitral area under changing hemodynamic conditions.

Storage, ultrastructural targeting and function of toposomes and hyalin in sea urchin embryogenesis.

This study compares by immunogold labeling the ultrastructural localization of a hexameric 22S glycoprotein, called toposome, with that of hyalin in unfertilized eggs and cells of hatched sea urchin blastulae. Nearly all hyalin is present in the electron translucent compartment of the cortical granules and in the translucent non-cortical pigment granules. In the blastula both of these intracellular stores have vanished and hyalin now forms a broad band below the apical lamina. By contrast, in the egg toposomes are present on the surface, as well as stored in yolk granules and in the electron dense lamellar compartment of the cortical granules. In the hatched blastula, toposomes that have been modified by limited proteolysis in the yolk granules, are associated with the plasma membranes of all newly formed cells, while the toposomes originating from the cortical granules have been incorporated as unmodified 160 kDa polypeptides into an extracellular double layer enveloping the embryo on the outside of the hyaline layer. From evidence discussed in detail, we conclude that the extracellular toposomes rivet the apical lamina to the surface and underlying cytoskeleton of the microvilli, while the modified toposomes from the yolk granules are responsible for position specific intercellular adhesion as they are released to the surface of newly formed cells. We propose that all the material stored in yolk granules is utilized for the assembly of new membranes.

Functional characterization of toposomes from sea urchin blastula embryos by a morphogenetic cell aggregation assay.

This paper documents the evidence that the large oligomeric glycoprotein complexes of unknown function first isolated as 22S particles from sea urchin embryos are the sole agents responsible for the adhesive integrity of sea urchin blastula embryos. The conclusion rests on the demonstration that polyclonal IgG (as serum or monovalent Fab) against whole membranes or butanol-solubilized components of membranes, as well as against the purified particle itself, completely blocks reaggregation of dissociated blastula cells and that this inhibition is reversed by neutralization of the inhibitory antibodies with purified 22S antigen. An essential aspect of the evidence is the combination of quantitative endpoint titrations in microtiter wells with the qualitative parameters of morphogenesis. The new data complement previous evidence that morphogenesis is mediated by a general class of particles, toposomes, responsible for mechanical linkage between cells and their positional guidance in embryogenesis.

Sequencing the human genome.

The title of the report by R. Myerowitz and N. Hogikyan on page 1646 of the issue of 27 June was incorrect. It should have been "Different mutations in Ashkenazi Jews and non-Jewish French Canadians with Tay-Sachs disease."

Characterization of toposomes from sea urchin blastula cells: a cell organelle mediating cell adhesion and expressing positional information.

Cell adhesion in the sea urchin blastula is mediated by a 22S genus-specific glycoprotein complex consisting initially of six 160-kDa subunits that are processed proteolytically as development proceeds. Noncytolytic removal of the 22S particle from the surface with either 2.5% butanol or trypsin renders dissociated cells reaggregation incompetent, and addition restores reaggregation and development. Polyclonal antibodies against the 22S complex prevent reaggregation in a genus-specific manner while monoclonal antibodies stain cell surface structures in a pattern consistent with a code that specifies the position of a cell in the embryo by a unique combination of subunits in its cell adhesion particles. The existence of similar particles in Drosophila and amphibian embryos suggests that these glycoprotein complexes are a general class of organelles, the toposomes, that in the embryo mediate cell adhesion and express positional information.

How to shape incentive plans for the hospital's top executives.

Surprisingly few hospital boards use incentive payment plans for their top executives. In this article, the author provides a rationale for their use, reviews various incentive plan formats, and suggests a sample incentive contract.

Interaction of D-LSD with blood platelets of rabbits: shape change and specific binding.

In blood platelets of rabbits, the shape change-inducing effect of D-lysergic acid diethylamide (D-LSD) has been compared with the D-LSD-binding. D-LSD, but not L-LSD, caused a shape change reaction (EC50 1.3 x 10(-9) M) which was inhibited by various 5-HT antagonists (methergoline and neuroleptic drugs), butaclamol showing marked stereospecificity. Strong inhibitors of 5 HT uptake were only weak in counteracting the D-LSD-induced shape change. Furthermore, D-[3H]LSD bound to platelets at a single saturable, high affinity, stereoselective site [Kd = (31.1 +/- 3.3) x 10(-9) M; Bmax = 28.9 +/- 2.7 fmols per 10(8) platelets]. This binding was strongly antagonized by D-LSD and methergoline and less by the hallucinogenic drugs, psilocin, bufotenin and N'N'-dimethyltryptamine. L-LSD an 5 HT, inhibitors of 5 HT uptake and neuroleptics, especially spiroperidol and butaclamol, had only a weak antagonistic effect. The latter showed stereospecificity. It is concluded that 1) the shape change reaction caused by D-LSD in platelets is mediated by the specific 5 HT-receptor, 2) the sites mediating the D-LSD-induced shape change reaction are not identical with those responsible for D-[3H]LSD-binding and both these sites are different from the 5 HT-transport sites and 3) with respect to D-LSD-binding sites, platelets and neurons are not exactly identical.

Inverse affects on thymidine incorporation in dissociated blastula cells of the sea urchin Paracentrotus lividus induced by butanol treatment and fab addition.

When dissociated blastula cells of the sea urchin Paracentrotus lividus were prevented from aggregating by extracting them with 2.5% n-butanol in sea water, they became unable to incorporate labelled thymidine, like over-diluted dissociated cells. If, however, cells were prevented from aggregating by the addition of Fab against plasma membranes, thymidine incorporation was not blocked, but increased compared to that of spontaneously aggregating cells. At the same time amino acid incorporation also increased.

Reconstitution of membranes and embryonic development in dissociated blastula cells of the sea urchin by reinsertion of aggregation-promoting membrane proteins extracted with butanol.

Blastula embryos of the sea urchin Paracentrotus lividus, when dissociated into single cells by exposure to Ca2+- and Mg2+-free sea water, reassociate spontaneously to form aggregates capable of development to the final larval form (pluteus). This aggregation is prevented by Fab fragments obtained by immunization with purified membranes from blastula embryos. The inhibition was reversed by soluble proteins extracted with butanol from purified membranes or from intact cells. These extracts also strongly stimulated the rate of reaggregation of dissociated cells in the absence of Fab fragments. Exposure of dissociated cells to 2.5% (vol/vol) butanol removed completely the protein(s) responsible for reaggregation of the cells without impairing their viability. Reaggregation and embryonic development were completely restored to the extracted cells by readdition of the proteins extracted from either membranes or cells. Extracted cells from Paracentrotus could be reconstituted with proteins from Arbacia.