Spectral features of amines of 2-phenylbenzazoles upon interaction with DNA

Absorption and fluorescent spectra of various synthetic aminophenyl derivatives of benzoxazole, benzothiazole and benzimidazole have been studied to estimate the efficiency of their binding with DNA. The significance of different functional groups of the fluorochromes for their interaction with DNA was determined, and main demands are formulated to the compounds to be used as potential fluorescent probes for DNA studies.     

Cyclic analogs of des-Arg9-[Leu8]bradykinin

Four cyclic derivatives of des-Arg9[Leu8]bradykinin have been obtained by classical methods of peptide chemistry. They are cyclo-(-X-Arg-Pro-Pro-Gly-Phe-Gly-Pro-Leu-), where X=Lys or none, and cyclo-(Y-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Leu-), where Y= Lys or Orn. Peptide bonds have been formed by the pentafluorophenylester method, and cyclization has been carried out in a diluted dioxane solution with 40% yield. Subsequent cleavage of protecting groups was made by treatment with hydrogen fluoride. The products obtained were purified by droplet counter-current chromatography. These substances liberate histamine from the rat mast cells comparably to bradykinin and fail to produce myotripic and vascular effects.     

Isolation and physico-chemical properties of a protein, included in an endotoxin from Yersinia pseudotuberculosis

A major protein of the endotoxin from Yersinia pseudotuberculosis was isolated from the complex lipid A--protein by treatment with SDS and triton X-100 followed by gel-chromatography on Sephacryl S-300. Protein has apparent molecular mass 40 kDa and alanine as N-terminal amino acid residue. CD and IR spectroscopy conformational changes of the protein molecule in the process of its isolation. The thermal and pH stabilities of the protein were investigated by the methods of intrinsic fluorescence and differential scanning microcalorimetry. The isolated protein revealed two thermal transitions (at 30-35 and 50-55 degrees C), which depend on Ca2+ concentration.     

A numerical model of expired flow in a monoalveolar lung model subjected to pressure ramps

A numerical investigation of pulmonary flow properties was carried out in a monoalveolar model composed of a balloon and a compliant tube in series, subjected to pressure ramps. The flow is shown to become quickly limited by a wave-speed mechanism, occurring at the peak flow. The critical point then travels upstream, while the main part of the exit flow rate is provided by the tube collapse. After the critical flow period, the flow becomes subcritical and viscous effects are predominant in the deeply collapsed tube.     

A simple and sensitive spectrophotometric method for the quantitative determination of solid supported amino groups

A simple and sensitive method for the quantitative determination of free amino groups on solid support is described. This approach is a modification of Ngo's [(1986) J. Biochem. Biophys. Methods 12, 349-354] method reported earlier. The method is based on the reaction of the solid support with an excess of 5'-O-(4,4'-dimethoxytrityl)-thymidine-3'-O-(2,4-dinitrophenyl) succinate (DTDS) in the presence of a catalytic amount of 4-dimethylaminopyridine. After removing the excess reagent, solid support is treated with perchloric acid to release 4,4'-dimethoxytrityl cation into the solution. The released 4,4'-dimethoxytrityl cation, which has a strong absorption at 498 nm (epsilon 498 = 70,000), is then determined spectrophotometrically. A comparative study of DTDS, N-succinimidyl-3-(2-pyridyldithio)propionate and 4,4-dimethoxytrityl chloride is also included. The method was found to be very useful to determine those amino groups which are available for functionalization of solid supports, especially, monitoring the functionalization of solid supports for affinity chromatography and synthesis of biopolymers.     

Larval and adult morphology ofCamallanus anabantis Pearse, 1933 andC. kulasirii (Yeh, 1960) (Nematoda: Camallanidae) from freshwater fishes,with notes on the validity of some related forms

Larvae and adults ofCamallanus anabantis andC. kulasirii, recovered from the West Bengali freshwater fishes,Anabas testudineus andOphicephalus punctatus, respectively, are described on the basis of detailed morphological studies under the light microscope. Larval forms collected fromA. testudineus are deemed to be of the third and fourth stages when compared with those from experimental studies of the life cycle ofC. anabantis. Moreover, the present fourth stage female larvae are similar to the females ofC. pearsei, both morphologically and metrically.C. pearsei is, therefore, believed to represent the fourth stage female larvae ofC. anabantis. Similarly, adult males and females recovered fromO. punctatus closely resembleC. kulasirii andC. fernandoi, respectively. The larval forms from this host are fourth stage and can be distinguished as males and females, but both possess a buccal capsule bearing beaded longitudinal ridges similar to that of adult males. The late fourth stage (just prior to the final moult) female larva is, however, found to possess a buccal capsule transitional between that of the adult male and female and also betweenC. kulasirii andC. fernandoi. C. fernandoi is, therefore, presumed to represent the females ofC. kulasirii. However,C. pearsei andC. fernandoi are regarded, for the present, asspecies inquirendae.     

Comparison of helodermin, VIP and PHI in pancreatic secretion and blood flow in dogs

Helodermin, VIP and PHI, which share a high degree of homology with secretin, have been identified in the gut but their physiological role is unknown. In this study 3 series of tests were carried out to determine the actions of helodermin, VIP and PHI on pancreatic secretion in 6 conscious dogs and amylase release from the dispersed canine pancreatic acini and to correlate the alterations in pancreatic secretory and circulatory effects in 24 anesthetized dogs. Helodermin, VIP and PHI infused i.v. in graded doses (12.5-200 pmol/kg.h) resulted in a dose-dependent increase in pancreatic HCO3 secretion reaching, respectively, 100%, 7% and 2% of secretin maximum. When combined with constant dose infusion of CCK-8 (100 pmol/kg.h), helodermin but not VIP or PHI augmented dose-dependently the HCO3 secretion. When added in various concentrations (10(-10)-10(-5)M) to the incubation medium of dispersed pancreatic acini only helodermin but not VIP or PHI increased dose-dependently amylase release reaching about 50% of CCK-8 maximum. In anesthetized dogs, the pancreatic blood flow (PBF) measured by electromagnetic blood flowmetry showed an immediate and dose-dependent increase following the injections of various doses of helodermin, VIP, PHI and secretin, the peak blood flow preceding by about 1 min the peak secretory stimulation. This study shows that helodermin resembles secretin in its potent pancreatic HCO3 stimulation but differs from VIP or PHI which are poor secretagogues but potent vasodilators. We conclude that if tested peptides are released in the gut, helodermin, like secretin, may be involved in the hormonal stimulation of exocrine pancreas, whereas VIP and PHI may serve mainly as vasodilators in the pancreatic circulation.     

Epithelial differentiation in Drosophila pupae

The construction of cell hairs on the wings in developing pupae of Drosophila provides a unique system for studies of the regulation of differentiation in the absence of cell division. Early steps in hair construction are the extrusion of cell hairs and the deposition of the external impervious layer called "cuticulin." Some properties of six of the most abundant proteins that are present during the early stages of hair construction are described. These proteins make up about 40% of the total protein of the preparation.     

On the subcellular localization of the polyamines

Putrescine, spermidine and spermine were determined in the nuclear fraction of rat liver which was obtained by density gradient centrifugation in non-aqueous media, i.e. under conditions which avoid migration of water-soluble compounds. Calculations of the distribution of the polyamines between nuclear and extranuclear compartments were based on the assumption that the DNA is concentrated in the nuclei. No significant losses of the polyamines occurred during fractionation. From the polyamine determination in tissue and nuclear fraction it appeared that 16-17% of the liver spermidine and spermine, and about 8% of the putrescine content was localized in the nuclei. The spermidine/spermine-ratios in nuclei and whole tissue were not significantly different. Pretreatment of the animals with inhibitors of ornithine decarboxylase caused a decrease of putrescine exclusively in the extranuclear compartments, in agreement with a higher proportion of the inhibitors in the cytoplasm. Since the nuclear volume of rat liver corresponds to about 5% of total liver volume, the concentration of spermidine and spermine is higher in the nucleus than in extranuclear compartments. Published histochemical localizations of the polyamines suggested very low polyamine concentrations in the nuclei of non-dividing liver and HeLa cells, but dramatic polyamine accumulations in metaphase and anaphase nuclei. These results are in disagreement with previously reported autoradiographic data, subcellular localizations based on density gradient centrifugations, and with our present results. Since subcellular localization is a key issue in all attempts to clarify cellular functions of the polyamines the careful revision of the techniques involved in subcellular polyamine localizations seems imperative.     

Characteristics of the differentiation of hematopoietic stem cells of the bone marrow in experimental lesions of the liver

The liver lesion in the CBA mice has been induced by administration of one of three agents five times every day; gamma-globulin fraction of antihepatocytotoxic serum in doses of 4.8 and 7.7 mg of protein per 100 g of body mass; gamma-globulin fraction of normal rabbit serum and bovine serum albumin in a dose of 4.8 mg of protein; three- four- or five-fold introduction of carbon tetrachloride in a dose of 0.5 ml per 100 g of body mass with oil (1:1) each three days; calibrated stenosis of the portal vein was produced. The total number of hemopoietic stem cells in the bone marrow was estimated by the colony-forming unit/spleen assay. Histological analysis of the colony-forming units was applied. The liver lesion was accompanied by a decrease in the ratio of the erythroid/granulocytic colonies.