Cryofractographic study of intercellular junctions in the populations of agar-cultivated Bordetella pertussis

The characteristic feature of replicas obtained from the freeze-fractures of B. pertussis unfixed cultures developing on casein charcoal agar for 1-7 days is the associative growth of highly polymorphic cells, ensured by the ramified system of intercellular connections (IC) formed by the derivatives of the outer layers of the cell wall. This proves that the associative location of bacterial cells, linked by numerous IC, in the preparation is not the artefact appearing in the process of their chemical fixation. In replicas obtained from the freeze-fractures of B. pertussis cultures, previously fixed with glutaraldehyde, osmic acid and uranyl acetate, oval cells with the cytoplasm having a relatively homogeneous structure and with the smoothed-out three-layer cell wall prevail. As a rule, IC are limited to the sites of direct contacts between individual cells.     

Cellular ultrastructure of the meningococcus when incubated in a continuous culture of human FL-line amnion

Some details of the ultrastructure of several meningococcal strains having had contacts with cells in continuous human amnion cell culture FL for 6 hours to 2 days have been defined with greater precision by means of electron microscopy. The study has shown that the contact of meningococci with the tissue culture is accompanied by the appearance of meningococcal forms with the defective cell wall, similar to L-forms: spheroplast, protoplast, gigantic cells and microcells, as well as budding variants. The meningococcal variants with the defective cell wall, appearing in the cell culture, and the forms occurring (in different proportions) in "ripe" meningococcal populations developing in the culture media for a long time and isolated from a human body have been found to have no essential differences in their fine structure. These data indicate that any external influences (meningococci are highly sensitive to such influences) produce sufficiently rapid changes, similar to L-transformation, in the fine structure of these microorganisms.     

Ultrastructure of the meningococcus in logarithmic and stationary development phases

The complex study of Neisseria meningitidis cultures A-208 in the time course of their development has disclosed that broth cultures in the logarithmic and stationary phases of their development are most valid on account of all their biological properties (the specific character of the reaction of agglutination, viability, the morphology of colonies and cells in light and electron microscopy). The use of scanning electron microscopy has made it possible to reveal bubbly endotoxin excretion in N. meningitidis and funnel-shaped depressions on their surface corresponding, probably, to nucleoid epicenters . In ultrathin sections some previously unknown features of the ultrastructure of N. meningitidis in the logarithmic and stationary phase of their development have been detected: (a) the morphological heterogeneity of N. meningitidis represented by cells of the "light" (L) and "dark" (D) types; (b) the surface structures of meningococcal cells from the cultures in the stationary phase of development show the tendency to smoothing out, which is accompanied by their sharper differentiation.     

Abnormal methylation of p16/CDKN2A AND p14/ARF genes GpG Islands in non-small cell lung cancer and in acute lymphoblastic leukemia

Multiplex methylation-sensitive PCR and methylation-specific PCR were employed in studying the methylation of CpG islands in the p16/CDKN2A and p14/ARF promoter and the first exon regions in non-small cell lung cancer (54 samples) and acute B-cell lymphoblastic leukemia (61 samples). Differences in methylation were detected between types of neoplasia as well as between CpG islands studied within the same types of tumors. High level of the p16/CDKN2A first exon CpC island methylation was revealed in non-small cell lung cancer (68%) and in acute B-cell lymphoblastic leukemia (55%) and the CpG island of p14/ARF first exon was nonmethylated in these types of tumors. The methylation of CpG-rich fragments of genes p16/CDKN2A and p14/ARF promoters was analysed. As was found out, CpG islands located in 5' areas of one and the same gene can differ in methylation frequencies. The comparison of sensitivity between methylation-specific PCR and methylation-sensitive PCR used in the methylations studies was carried out.     

Metabolism of the androgenic precursor androstenedione and effects of aromatase inhibitors in cultured blood lymphocytes

Blood lymphocytes from 26 women were cultivated in RPMI-1640 for 72 hrs with or without dexamethasone, dbcAMP, and TPA. Androstenedione conversion was most often activated by the dbcAMP, then by Dex and TPA. Response to dbcAMP was most obvious in women under 50, whereas response to Dex--in women over 50. Aromatase inhibitor fadrosol displayed a tendency towards prevention of the activation of androstenedione conversion in cultivated lymphocytes. A possibility of aromatase gene induction in cultivated lymphocytes cannot be completely excluded and demands further investigation with the aid of molecular-genetic methods.     

Noninvasive estimation of the oxygen state of experimental tumor by diffuse optical spectroscopy

The potentialities of diffuse optical spectroscopy for the noninvasive estimation of the oxygen state of experimental tumors have been demonstrated. The distribution of total, oxygenated and deoxygenated hemoglobin, as well as the level oxygen saturation of blood have been shown using two tumor models differing in the histological structure and functional characteristics. The results obtained by the optical method have been verified by immunohistochemical examination of tissue specimens with the exogenous hypoxia marker pimonidazole.     

Evolutionarily-conserved gene CKAP2,located in region 13q14.3 of the human genome, is frequently rearranged in various tumors

The human CKAP2 gene, which is involved in diffuse large B-cell lymphomas, was localized via screening the GeneBridge 4 somatic cell radiation hybrid panel by means of the polymerase chain reaction (PCR). The CKAP2 gene was mapped between the WI-15460 and WI-3673 markers at the boundary between regions 13q14.3 and 13q21.1, at the distance of 14.39 cR (with 4.8 cR per cM) from the WI-5867 framework marker (lod score > 2.26). The human CKAP2 gene displayed high homology to mouse and rat expressed orthologs, A CKAP2-like sequence was found in human chromosome 14 and assumed to be a pseudogene resulting from duplication and subsequent mutations of the CKAP2 gene on chromosome 13. A possible role of the CKAP2 gene in oncogenesis associated with deletions and rearrangements of region 13q14.3-21.1 is discussed.     

New steroid glycosides from the starfish Fromia milleporella

Two new steroid glycosides from the starfish Fromia milleporella collected in the Seychelles were isolated and characterized: milleporoside A, (20R, 24R)-29-O-[3-O-methyl-beta-D-xylopyranosyl-(1-->4)-3-O-methyl-beta-D-xylopyranosyl]-24-ethyl-5alpha-cholestane-3beta,4beta,6alpha,8,15beta,16beta,29-heptaol, and milleporoside B, (20R, 24R)-(22E)-28-O-[3-O-methyl-beta-D-xylopyranosyl-(1-->4)-3-O-methyl-beta-D-xylopyranosyl]-24-methyl-5alpha-cholest-22-ene-3beta,4beta,6alpha,8,15beta,16beta,28-heptaol. The structures of the glycosides were determined from their spectra and a comparison with spectral characteristics of known compounds. These compounds exhibit a moderate cytostatic activity toward the embryos of the sea urchin Strongylocentrotus intermedius.     

Microbiological and production characteristics of the high-temperature Kongdian bed revealed during field trial of biotechnology for the enhancement of oil recovery

Microbiological technology for the enhancement of oil recovery based on the activation of the stratal microflora was tested in the high-temperature horizons of the Kongdian bed (60 degrees C) of the Dagang oil field (China). This biotechnology consists in the pumping of a water-air mixture and nitrogen and phosphorus mineral salts into the oil stratum through injection wells in order to stimulate the activity of the stratal microflora which produce oil-releasing metabolites. Monitoring of the physicochemical, microbiological, and production characteristics of the test site has revealed large changes in the ecosystem as a result of the application of biotechnology. The cell numbers of thermophilic hydrocarbon-oxidizing, fermentative, sulfate-reducing, and methanogenic microorganisms increased 10-10 000-fold. The rates of methanogenesis and sulfate reduction increased in the near-bottom zone of the injection wells and of some production wells. The microbial oil transformation was accompanied by the accumulation of bicarbonate ions, volatile fatty acids, and biosurfactants in the formation waters, as well as of CH4 and CO2 both in the gas phase and in the oil. Microbial metabolites promoted the additional recovery of oil. As a result of the application of biotechnology, the water content in the production liquid from the test site decreased, and the oil content increased. This allowed the recovery of more than 14000 tons of additional oil over 3.5 years.     

A study of the inhomogeneity of bilayer lipid membranes by measurements of higher harmonics of transmembrane current

Higher harmonics of alternating current in bilayer lipid membranes caused by sinusoidal voltage applied to the membrane were measured. The bilayer lipid membranes were prepared from diphytanoylphosphatidylcholine in n-decane and n-tetradecane, and measurements were conducted with the aid of an analog-to-digital converter of 16th category. Sinusoidal voltage was formed using a digital-to-analog converter of the 16th category. The dynamic region of measurements was up 90 dB. The results of measurements were used to determine the alpha and beta coefficients of the expansion of membrane capacity C in terms of membrane voltage U C = C0 (1 + alphaU2 + betaU4). We showed in the framework of the electrostriction model that the relation between the alpha and beta coefficients characterizes the inhomogeneity of bilayer lipid membrane with respect to its thickness and Young modulus of elasticity.