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.     

Eicosanoids in insect immunity: bacterial infection stimulates hemocytic phospholipase A2 activity in tobacco hornworms

Intracellular phospholipase A(2) (PLA(2)) is responsible for releasing arachidonic acid from cellular phospholipids, and is thought to be the first step in eicosanoid biosynthesis. Intracellular PLA(2)s have been characterized in fat body and hemocytes from tobacco hornworms, Manduca sexta. Here we show that bacterial challenge stimulated increased PLA(2) activity in isolated hemocyte preparations, relative to control hemocyte preparations that were challenged with water. The increased activity was detected as early as 15 s post-challenge and lasted for at least 1 h. The increased activity depended on a minimum bacterial challenge dose, and was inhibited in reactions conducted in the presence of oleyoxyethylphosphorylcholine, a site-specific PLA(2) inhibitor. In independent experiments with serum prepared from whole hemolymph, we found no PLA(2) activity was secreted into serum during the first 24 h following bacterial infection. We infer that a hemocytic intracellular PLA(2) activity is increased immediately an infection is detected. The significance of this enzyme lies in its role in launching the biosynthesis of eicosanoids, which mediate cellular immune reactions to bacterial infection.     

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.     

The connection of visual evoked potentials with the subjective differences between emotional expressions of the "schematic face"

Human cortical visual potentials (VEP) were studied to obtain electrophysiological data concerning face discrimination and to compare them with the direct estimates of differences between faces obtained in the previous publications. The present schematic faces varied in curvature of a mouth and/or declination of eyebrows. These features determined the emotional expression of the schematic faces. We recorded the VEP as the response to the instant replacement of one schematic face (referent stimulus) by an other one (test stimulus) rather then to presentation of a single stimulus. Thus we recorded direct electrophysiological differences between schematic faces. A characteristic feature of this approach was the application of the set of functionally connected test stimuli with monotonously increasing values of differences between the referent and test stimuli. In a result of analysis the complex of components P120-N180-P230 in sites O1, O2, P3, P4, T5, T6 was described. Interpeaks amplitudes of the components shows high correlations with subjective differences between the same pairs of stimuli as well as with physical (configurative) differences between stimuli measured as the angles of lines, defining curvature of a mouth and a declination of eyebrows. The highest correlation with subjective estimates of emotional differences between faces was shown by interpeaks amplitudes N180-P230 in sites O1 and P3. In the some time the interpeaks amplitudes P120-N180 in sites O1 and T5 reflected highest correlation between configurative measures and subjective estimates of stimuli differences.