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Biomedical subjects

D Ostrovsky

Publications and source records attributed to D Ostrovsky.

At least 19 recordsLinked to original sources

Effect of oxidative stress on the biosynthesis of 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate and isoprenoids by several bacterial strains.

In this study, the gram-negative bacteria Xanthomonas campestris, Xanthomonas maltophilia, and Pseudomonas putida, facultative parasites of plants and animals, were shown to accumulate 2-C-methyl-d-erythritol-2,4-cyclopyrophosphate (MEC) in response to benzyl-viologen-induced oxidative stress. Corynebacterium ammoniagenes mutants capable of accumulating MEC in the absence of an exogenous oxidative stress inducer were obtained. Isoprenoid synthesis and MEC synthesis in these and other bacteria were shown to be alternative processes, while biosynthesis of brominated polyene xanthomonadin (an antioxidant pigment of X. campestris) increased concomitantly with the accumulation of MEC.

Carotenoids↗

Long-term patient self-management of oral anticoagulation.

BACKGROUND: The management of oral anticoagulation is fraught with difficulties. This study assessed a new model of anticoagulation management regarding the ability, safety, and efficacy of patients to self-monitor and self-adjust the dose of their oral anticoagulants guided by a capillary whole-blood prothrombin time (PT) monitor. METHODS: This investigation is a retrospective cohort study of 20 patients compared with 20 matched control patients receiving oral anticoagulation at a tertiary medical institution. RESULTS: Study patients monitored their PTs 2153 times during a mean interval of 44.7 months compared with 1608 PTs in matched control patients during a mean interval of 42.5 months. Study patients made an average of 11.5 dosage changes per patient, contrasted with 22.7 changes per control patient (P < .001). The PTs in study patients were within the recommended therapeutic range in 88.6% (95% confidence interval, 87.2 to 89.9) of the determinations compared with 68.0% (95% confidence interval, 65.7 to 70.3; P < .001) of the determinations made by the matched control patients. In response to the 2153 PTs, study patients made 67 (3.1%) dosage decisions that were considered incorrect based on physician guidelines. None of these changes led to adverse outcomes. There was no significant difference in complication rates between the two groups. CONCLUSIONS: Results from what is the first long-term study of patient self-monitoring of PTs and self-adjustment of the warfarin sodium dosage for oral anticoagulation suggest that patients can successfully measure their own PTs, adjust their own warfarin dosage, and achieve a degree of therapeutic effectiveness at least as good, if not better than patients managed in an anti-coagulation clinic. Larger, prospective, randomized trials are needed to confirm the efficacy and safety of this new approach to therapy and to assess its cost-effectiveness.

Administration, Oral↗

Heat treatment of Corynebacterium ammoniagenes leads to aeration dependent accumulation of 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate.

2-C-methyl-D-erythritol-2,4-cyclopyrophosphate (MEC) identified as a new bacterial oxidative stress substance (Ostrovsky D. et al. (1993) Biochem. J., 295, 901-902) was shown to accumulate in Corynebacterium (Brevibacterium) ammoniagenes cells aerobically cultivated in peptone-yeast extract-glucose broth on heating for 1 hour at 45 degrees C. The enzyme(s) responsible for MEC biosynthesis is evidently oxidized for activation and is completely loosing its activity on anaerobic incubation at this temperature in an hour. Salt stress or drying did not provoke the MEC biosynthesis.

Aerobiosis↗

Bacteria and pesticides: a new aspect of interaction--involvement of a new biofactor.

Positively charged hydrophobic pesticides of the dipyridyl family [diquat, paraquat, benzylviologen (BV++), etc.] were shown to provoke accumulation of 2-methylbutane-1,2,3,4-tetraol-2,4- cyclopyrophosphate in the cells Corynebacterium (Brevibacterium) ammoniagenes while neutral dipyridyls were not. Hydrophobicity was also an important factor in this phenomenon. Of the other pesticides tested, only linuron was effective. BV++ also induced biosynthesis of the compound in Rhodococcus rhodochrous, Rh.ruber, Rh.sp. (Nocardia corynebacteroides). These microorganisms as well as most of the previously identified oxidative stress activated producers of this new cyclopyrophosphate were able to synthesize free radical generating compounds. The microorganisms concerned belong mainly to the order Actinomycetales.

2,2'-Dipyridyl↗

Bacterial oxidative stress substance spontaneously recyclizes to form 2-methylbutane-1,2,3,4-tetraol-1,2-cyclophospho-4-phosphate.

The cells of Corynebacterium (Brevibacterium) ammonia-genes cultivated in a medium supplemented with diquat or benzylviologen accumulate 2-methylbutane-1,2,3,4-tetraol-2,4- cyclopyrophosphate as revealed by 31P-NMR spectroscopy. On heating at 120 degrees C for 30 min the cells still maintain a substantial portion of this compound and acquire new cyclic phosphates characterized by 31P-NMR chemical shifts of +17.3 and +20 p.p.m. The +17.3 p.p.m. component was isolated from the preparation of the purified cyclopyrophosphate kept for some time at pH above 7 and it was shown to be 2-methylbutane-1,2,3,4-tetraol-1,2,- cyclophospho-4-phosphate on the grounds of two-dimensional NMR spectroscopy.

Benzyl Viologen↗

A new cyclopyrophosphate as a bacterial antistressor?

In a number of bacteria an unusual glycosyl pyrophosphate (31P NMR signal chemical shift at about -15 ppm) was detected when the cells were subjected to oxidative stress. This substance from Brevibacterium ammoniagenes has now been identified as 2-methyl-butan-1,2,3,4,-tetraol-2,4-cyclopyrophosphate, which is accumulated in the cell under certain conditions in concentrations of of about 50 mM. It is now suggested that this compound is the long sought after bacterial antistressor.

Brevibacterium↗

The ability of bacteria to synthesize a new cyclopyrophosphate correlates with their tolerance to redox-cycling drugs: on a crossroad of chemotherapy, environmental toxicology and immunobiochemical problems.

Many redox-cyclers were recently shown to induce, in some bacterial species, large-scale biosynthesis of a new 2-methylbutan-1,2,3,4-tetraol-2,4-cyclopyrophosphate believed to be involved in anti-stress reactions. In the present study Mycobacterium smegmatis, Micrococcus luteus and Brevibacterium ammoniagenes were shown to begin synthesis of the new cyclopyrophosphate when cultivated in a medium containing furacilin or furadonin (widely used nitrofuran antibacterial drugs) and to maintain close to normal growth rates, whereas Staphylococcus aureus, Bacillus subtilis and Escherichia coli were inhibited by the drugs and were unable to synthesize the cyclopyrophosphate compound. Preferential binding of Mg2+ and Cd2+ with one or other phosphoryl groups of the cyclopyrophosphate, which was indicated by selective changes of 31P-NMR chemical shifts and intramolecular hydrogen bonding, is suggested as a reason for this selectivity.

Bacteria↗

Synthesis of a new organic pyrophosphate in large quantities is induced in some bacteria by oxidative stress.

Brevibacterium ammoniagenes and Micrococcus luteus were shown to synthesize up to 50 mM of a novel substance, 2-methylbutan-1,2,3,4-tetraol 2,4-cyclopyrophosphate, in response to oxidative stress created by benzyl viologen and other redox mediators under aerobic conditions. The substance, which represents greater than 50% of the extractable phosphorus, is suggested to play a role as a bacterial antistressor and is thought to be a product of condensation of two molecules of phosphoenolpyruvate whose accumulation is prompted by conversion of intracellular NADPH into an oxidized form.

Brevibacterium↗

Interconversion of radical and nitrone forms of lysodektose--a new trisaccharide from Micrococcus lysodeikticus.

Isolated from Micrococcus lysodeikticus, 6-O-(2-deoxy-2-(N-methyl)hydroxilamino-beta-D-glucopyranosyl)-alph a-alpha- trehalose (lysodektose) is oxidized by K3Fe(CN)6 in a stepwise manner to become a nitroxyl radical and a nitrone with a double bond in the fragment O-N = CH2 which could be reduced to the original hydroxylamine form with sodium borohydride. Thus derivatives of lysodektose specifically labelled with 2H and (or) 3H in the methyl group are easily obtained. When oxidized in cells poisoned with vitamin K analogues, lysodektose is transformed into nitrone concomitant with modification of its methyl group. Participation in the antioxidant defence of the bacteria is suggested for this new trisaccharide.

Carbohydrate Sequence↗

Migration of chick blastoderm under the vitelline membrane: the role of fibronectin.

In the earliest stages of its development the chick blastoderm is a flattened disc at the surface of the yolk. It gradually increases in diameter, partially because the cells are rapidly proliferating, but also because the cells at the periphery (the margin of overgrowth) are migrating in a centrifugal direction. These cells utilize the inner surface of the vitelline membrane as their substratum. In the normal blastoderm, these cells at the edge of the spreading blastoderm are the only cells which are attached to the vitelline membrane. This investigation is concerned with the possible role played by fibronectin in the interaction between these migrating cells and the vitelline membrane. Chick blastoderms, explanted by the New (1955) technique have been treated with synthetic peptides that mimic the adhesive recognition signal of the fibronectin molecule. The pentapeptide GRGDS (containing the specific RGD cell adhesion sequence) caused the edge cells of the blastoderm to detach within minutes, and the expansion of the blastoderm was inhibited for about 4 hr. After this period there was gradual recovery and the cells reattached and spreading resumed. Examination of the margin of the blastoderm by scanning electron microscopy showed that cell processes were lost soon after treatment with GRGDS but concomitant with reattachment and the resumption of spreading, the cell processes reformed. The pentapeptide GRDGS (with the amino acids G and D inverted) produced a brief inhibition of spreading, but after an hour these blastoderms spread at the same rate as controls. Immunocytochemical staining with anti-fibronectin demonstrated that fibronectin was not only present at the interface of the edge cells and the vitelline membrane, but also between the epiblast and the hypoblast. These results indicate that tissue movement during blastoderm spreading is dependent upon fibronectin and that the specific RGD amino acid sequence, and presumably the VLA/integrin family of receptors, is involved in this embryonic morphogenetic movement.

Amino Acid Sequence↗

Somitogenesis in the mouse embryo.

This report describes the initiation of somitogenesis in the mouse embryo. Correlations are made with fibronectin distribution around the unsegmented mesoderm and the distribution of cytoskeletal elements within the cells as they undergo morphogenetic movements. The same temporal and topological changes in fibronectin, laminin, and cytoskeletal elements are seen in mouse somitogenesis as in the chick embryo. A notable exception is that the epithelial stage of somitogenesis in the mouse does not form a closed vesicle as it does in the chick. In the mouse the mesial portion of the forming somite does not become epithelial before the migration of sclerotomal cells.

Actins↗

Alpha actinin distribution and extracellular matrix products during somitogenesis and neurulation in the chick embryo.

A discrete stage in two different morphogenetic processes has been examined employing fluorescently labelled alpha-actinin as a probe to localize native alpha-actinin and antibodies to localize fibronectin and collagen type I. The stage of somitogenesis examined is the transition from the compact mesenchymal somitic mass to the epithelial somitic vesicle (ie, epithelialization of the somite). The stage of neurulation examined is the transition from the relatively flat neuroepithelium to the approximation of the neural folds. Before these morphogenetic movements begin, the neuroepithelium is sitting upon a basal lamina and interstitial collagen, and the somite is surrounded by a meshwork of interstitial collagen. During both of these processes, the cells become narrowed at their apices in the region of the tissue that is becoming concave, and alpha-actinin is localized in the apices. The localization of intracellular alpha-actinin and extracellular fibronectin, and the distribution of collagen, suggest that there is a coordinated appearance and distribution of these molecules that is temporally associated with these discrete morphogenetic events.

Actinin↗

On the role of fibronectin during the compaction stage of somitogenesis in the chick embryo.

During the early stages of somitogenesis in the chick embryo the presomitic cells in the segmental plate undergo compaction. The aggregation of segmental plate cells is stimulated by fibronectin. The stimulation of segmental plate cells to aggregate and undergo compaction can be effected in isolated segmental plate cells, in isolated segmental plates, and in intact embryos removed from the yolk. The fact that the segmental plate cells react with greater vigor to cellular fibronectin than to plasma fibronectin suggests a specific molecular mechanism in the initiation of somitogenesis.

Animals↗

Fibronectin distribution during somitogenesis in the chick embryo.

Somite formation in vertebrates is a multi-stage process. From a relatively homogeneous rod of mesenchyme, the segmental plate, somites are formed in a repeating sequence. Cell-cell adhesion has been proposed as a causal factor in somitogenesis. This led to an analysis of fibronectin in the segmental plate with respect to the initiation of somitogenesis. The pattern of fibronectin distribution can be correlated with the initiation of somitogenesis in the anterior portion of the segmental plate. Fibronectin distribution was determined using a high resolution antibody localization technique. Differences in fibronectin distribution were verified with computer-assisted image analysis. The evidence presented supports the hypothesis that an increase in cell-cell adhesion is a significant factor in the initiation of somitogenesis.

Animals↗

Light microscope observations on actin distribution during morphogenetic movements in the chick embryo.

The cellular distribution of actin during two morphogenetic processes in the chick embryo has been observed, using a high-resolution fluorescent technique, with heavy meromyosin as a probe. These cytoskeletal elements have been implicated in all cell and tissue movements in the embryo. It is now commonly accepted that microfilaments are necessary to provide the motive force for morphogenesis. Two morphogenetic movements in the early embryo have been studied at the light microscope level. During somitogenesis, the mesenchymal segmental plate becomes transformed into a meristic series of transient epithelial vesicles. Again, actin distribution is diffuse and random before the morphogenetic event. During epithelialization, actin becomes prominent in the apical regions of the epithelial cells. Cells in the somitic epithelial vesicles, the core cells, appear to be passive participants in this process, and consequently show no distinct cellular localization of actin.

Actins↗