Experimental studies of molecular interactions between nitrogen bases of nucleic acids.

New experimental results concerning molecular interactions between the nitrogen bases of nucleic acids in the crystalline phase and in vacuo are reported. The temperature dependence of the evaporation rate is measured for solid species. The sensitivity of conventional methods of sublimation heat measurements was improved essentially using a quartz resonator serving as a precise sensor of evaporation rate. Sublimation heats were found for both canonical bases and a number of their derivatives. The in vacuo formation of base associates interacting through hydrogen bonds was observed with a field mass spectrometer. The dimer formation enthalpies, which are indicative of a stronger attraction in complementary pairs compared with noncomplementary ones, were derived from the temperature dependence of ionic currents. Hydrogen-bound complexes of more intricate associates (base trimers and aqueous molecules associates) were studied. The energy gain in the formation of trimers of identical molecules was shown to be larger (per base molecule) than that for dimers.     

Studies of human liver bilirubin-glycosyl transferase. Bilirubin UDP-xylosyl and UDP-glucuronyl transferase activities in diseased human livers.

The activity of bilirubin UDP-xylosyl transferase as well as UDP-glucuronyl transferase in liver biopsy specimens of 3 control subjects, 42 cases with liver disease and 5 cases with Gilbert's syndrome was measured. Normal values of these enzyme levels were determined to be 142--302 U/kg protein for the former and 260--400 U/kg protein for the latter. Both enzyme levels in acute hepatitis in convalescence and chronic hepatitis were nearly in the normal range. In the cirrhotic liver they tended to a small decrease and patients with Gilbert's syndrome demonstrated significantly decreased enzyme levels. These enzyme levels were only correlated with serum unconjugated bilirubin concentration, but not with the other liver function tests. Finally, both enzyme activities were exactly correlated with each other.     

The correlation of ethanol-induced depression of glucose and water transport with morphological changes in the hamster jejunum in vivo

Glucose and water transport is depressed in the hamster jejunum in vivo by ethanol (4.8%) which also produced fluid-filled blebs at the tips of the villi. The epithelial cells over the blebs appeared stretched and cuboidal, the lateral intercellular spaces (LIS) were no longer recognizable, and the lacteals were closed. Forty-five minutes after discontinuation of the ethanol, water transport returned to normal while glucose transport remained depressed. At this time the villus structure had returned to normal. The blebs had disappeared, the LIS were again recognizable, and their appearance and number were similar to those in the control animals. Thus, the depression of water transport correlated with the obvious structural changes caused by ethanol; however, the depression of glucose absorption is associated with some effect of ethanol not evident by routine light microscopy.     

The formation of alpha-D-(1----3) branch linkages by a D-glucansucrase from Streptococcus mutans 6715 producing a soluble D-glucan

An exocellular D- glucansucrase that synthesizes a water-soluble, alpha-D-(1----6)-linked D-glucan having a high proportion of alpha-D-(1----3) branches was purified from the culture broth of Streptococcus mutans 6715. The rate of incorporation of D-[14C]glucose from [14C]sucrose into D-glucan of high molecular weight by this enzyme was increased (stimulated) by the presence of exogenous Leuconostoc mesenteroides B- 512F dextran, and it was found that this dextran could act as an acceptor. A highly branched dextran, containing 45-50% of alpha-D-(1----3) branch linkages, did not stimulate the enzyme nearly so much as B- 512F dextran, which has a low degree (5%) of alpha-D-(1----3) branches. We interpret this as evidence that the stimulating effects of dextran are not due to priming. If they were, the more highly branched dextran should have produced the greatest stimulation per unit weight, because a much greater number of nonreducing-end, priming sites would be available. We show that the D- glucansucrase was capable of transferring D-glucosyl groups from sucrose to B- 512F dextran to form alpha-D-(1----3) branches, thereby rendering the dextran more resistant to hydrolysis by endodextranase . The presence of 1.6M ammonium sulfate caused the enzyme to synthesize a D-glucan having a much higher percentage of alpha-D-(1----3) linkages.     

The mutagenic potencies of plant extracts containing quercetin in Salmonella typhimurium TA98 and TA100

Four commercial ethanolic plant extracts, Tinctura Alchemillae, Extractum Crataegi, Extractum Myrtilli and Tinctura Hyperici, were tested for their mutagenicity in Salmonella typhimurium TA98 and TA100 with and without S9 mix obtained from rats pretreated with phenobarbital. The extracts studied differed greatly in their mutagenic potencies but exhibited a very similar mutation pattern in which the strongest effect was always seen in tester strain TA98 with S9 mix. Simultaneously we investigated the extracts for the presence of quercetin and kaempferol. Only quercetin was detected in small amounts by thin-layer chromatography (TLC). The fractions containing quercetin were separated and collected using a Sephadex LH-20 column. Two different methods were employed to estimate the amount of quercetin in the extracts: a colorimetric assay developed by Christ and Müller, and a complexometric method by Belikov. The quercetin concentrations ranged between 2 mg (Tinctura Alchemilla) and 89 mg (Tinctura Hyperici) per 100 g of extract. We suggest that the mutagenicity of the 4 plant extracts is mainly due to the presence of quercetin for the following reasons: (1) all the plant extracts exhibit a mutation pattern which is very similar to that of quercetin, (2) the mutagenic potential of the extracts correlates well with their quercetin content, considering the fact that plant extracts are very complex mixtures often containing toxic or antimutagenic compounds.     

Loss of penicillinase plasmids of Staphylococcus aureus after treatment with L-ascorbic acid

When 2 clinical strains of plasmid-mediated penicillin-resistant Staphylococcus aureus were treated with 1 mM sodium ascorbate for 6 h, 12-35% colony-forming units (CFU) irreversibly lost their ability to produce beta-lactamase. Agarose gel electrophoresis showed the disappearance of the plasmid bands from the lysates of colonies in which susceptibility to penicillin was induced by ascorbate treatment. Since ascorbic acid is a compound that is completely non-toxic, the possibility of its therapeutic use in the treatment of multiresistant bacterial diseases is proposed.     

Quantitative histology of the seasonal changes in the gonads of summer whiting, Sillago ciliata

The histology of the gonads of free and captive summer whiting, Sillago ciliata , were compared to assess the potential of this fish for aquaculture. There were no qualitative differences between the free and captive males: both completed spermatogenesis through to spermiation. The captive females were non-reproductive, reaching only the vitellogenic stage before the eggs were resorbed by atresia.
A reproductive score was devised to assess reproductive potential in the comparison of the two populations. The females in the captive population had lower reproductive potentials, as shown by the score, and started development a month later than the normal free population. The captive males had higher reproductive scores than the free fish but they were also not spawning and, therefore, the higher score indicated that this potential was not realized. The score provides a standard against which experimental results can be compared and subsequently assessed.     

Further data on the development of SRIF-like immunoreactive nerve cell populations in the chick embryo brain stem. II. Midbrain tegmentum.

Further immunocytochemical analysis of the neuroblasts with SRIF-like immunoreactivity (ir) was carried out on the chick embryo midbrain tegmentum. 5 or 100 microns mesencephalon sections were obtained from 60 White Leghorn chick embryos at stages (E = Embryonic days) ranging from E4 1/2 to E18 and incubated with rabbit polyclonal antibodies against synthetic cyclic Somatostatin-14, according to PAP-DAB technique. In the midbrain tegmentum the ir appeared as from E12. From E12 to E13 1/2-E14 the ir distribution gradually changed. From E14 to E18 numbers and spatial arrangement of the positive neuroblast groups did not show substantial changes; in these respects the ir distributional pattern proved to be similar to the one observed by the Authors in adult animals. From E17 to E18 a decrease in the positive neuroblast density appeared to occur, particularly in a ventrally placed group. These results are consistent with a possible local regulative role of the SRIF.