Computer simulations of cyclic enkephalin analogues

Molecular dynamics simulations and energy minimization studies of cyclic enkephalin analogues incorporating retro-inverso modifications have been carried out. The dynamic trajectories are analyzed in terms of the relative mobility of the 14-membered rings, conformational transitions among equilibrium states, and hydrogen-bonding patterns. The cyclization of the molecules reduces the motion of the ring structures substantially. Time-correlated conformational transitions resulting in the reorientation of peptide units are observed. Hydrogen bonds form principally C7 structures. Because of the incorporation of retro-inverso residues, C6 and C8 structures are also formed. Starting conformations for energy minimizations were obtained from the molecular dynamics simulations and from a systematic search of the conformational space available to the molecules. Several minimum energy backbone and side-chain conformations were found for each analogue. The effect of retro-inverso residues on hydrogen-bonding patterns and backbone conformations is discussed.     

Chicken developmental antigens in 15I5-B-congenic lines

Six partially developed 15I5-B-congenic lines of chickens were used to assess the genetic influence on the developmental expression of selected epitopes of two avian developmental antigen systems: chicken fetal antigen (CFA) and chicken adult antigen (CAA). Both CFA and CAA are serologically and molecularly complex hematopoietic antigen systems, yet little is known about genetic influences on their expression. Using polyclonal rabbit anti-CFA, only slight variations in overall CFA expression on peripheral erythrocytes were observed during neonatal development; no consistent trend was evident. In contrast, analysis with monoclonal antibody 10C6 revealed that the incidence of CFA determinant 8 (CFA8) on erythrocytes of the early neonate was significantly reduced in line 15I5 compared with lines .6-2, .7-2 and .15I-5; line .C-12 also exhibited a reduced CFA8 incidence at hatching. Likewise, the CAA epitope detected by monoclonal antibody 3F12 was found to appear at a slower rate on erythrocytes from lines 15I5 and .C-12 than on those of other lines. Similar results were obtained using the anti-CAA monoclonal 4C2 where reduced expression was found in lines 15I5, .C-12, and .P-13. Results of complement-mediated cytolysis using the positive control 9F9 monoclonal antibody suggested that observed genetic differences were not due to inherent differences in erythroid cytolytic sensitivity. Neither could the results be explained by the incidence of circulating reticulocytes vs. mature erythrocytes within the lines. Rather, the results suggest that different genetic lines of chickens vary in the developmental kinetics of definitive erythrocyte subpopulations bearing specific phenotypes defined by monoclonal antibodies. These findings are discussed in light of previous observations using these B-congenic lines.     

Regulation of the manganese-containing superoxide dismutase is independent of the inducible DNA repair system in Escherichia coli

Studies on the induction of the manganese-containing superoxide dismutase in several strains of Escherichia coli with different mutations in recA and lexA revealed that the inductions of the Mn-isozyme and of the SOS system by oxygen free radicals are not coregulated. We also studied the synthesis of the manganese-superoxide dismutase in the temperature-dependent, protease-constitutive strain recA441(tif-1) that also contained a lac fusion in an SOS gene. A shift to the temperature at which recA441 has constitutive protease activity did not induce Mn-superoxide dismutase but did induce beta-galactosidase. The data clearly demonstrate that induction of the Mn-superoxide dismutase is independent of the SOS system.     

Location of polar substituents and fatty acyl chains on lipid A precursors from a 3-deoxy-D-manno-octulosonic acid-deficient mutant of Salmonella typhimurium. Studies by 1H, 13C, and 31P nuclear magnetic resonance

Eight anionic disaccharide precursors of lipid A accumulate at 42 degrees C in 3-deoxy-D-manno-octulosonic acid-deficient temperature-sensitive mutants of Salmonella typhimurium. These compounds comprise a series of lipids based on the minimal structure, O-[2-amino-2-deoxy-N2,O3-bis(3-hydroxytetradecanoyl)-beta-D-glucopyranos yl] -(1----6)-2-amino-2-deoxy-N2, O3-bis(3-hydroxytetradecanoyl)-alpha-D-glucopyranose 1,4'- bisphosphate (designated lipid IVA) that differ from each other by the presence of an additional phosphoethanolamine moiety (IIIA), or an aminodeoxypentose moiety (IIA), or both (IA). A homologous set of metabolites is further derivatized with a palmitoyl function; these are designated IVB, IIIB, IIB, and IB (Raetz, C. R. H., Purcell, S., Meyer, M. V., Qureshi, N., and Takayama, K. (1985) J. Biol. Chem. 260, 16080-16088). The attachment of the palmitoyl moiety, known to be on the reducing terminal GlcN residue by mass spectrometry, was determined to be O-beta of the N2-linked beta-hydroxymyristoyl group of that residue of IVB by 13C NMR and two-dimensional 1H chemical shift correlation spectroscopy experiments. 31P NMR indicated the presence of diphosphodiester moieties in IIIA, IIIB, and IA and monophosphodiester moieties in IIA and IA. Selective 1H decoupling of the 31P spectrum of IIIA demonstrated that the O-diphosphoethanolamine moiety is attached to the O4' position in IIIA. On the basis of the observed 31P chemical shifts it was concluded that the aminodeoxypentose is located at position 1 in IIA and IA, while diphosphoethanolamine is most likely located at O-4' in IA and IIIB, as in IIIA.     

Leishmania mexicana: purine-metabolizing enzymes of amastigotes and promastigotes

Cultured promastigote and isolated amastigote forms of Leishmania mexicana mexicana have been surveyed for the presence of enzymes involved in purine metabolism. Quantitative but not qualitative differences between the enzymes of two forms were discovered. There were found to be significant differences between the enzyme content of L. m. mexicana and that reported for L. donovani. Extracts of both parasite forms of L. m. mexicana were found to have higher levels of adenine deaminase (EC 3.5.4.2) and guanine deaminase (EC 3.5.4.3) than adenosine deaminase (EC 3.5.4.4). There appeared to be two distinct nucleosidases (EC 3.2.2.1), one active on nucleosides, the other on deoxynucleosides. Phosphorylase (EC 2.4.2.1) could be detected only in the catabolic direction. Nucleotidases were present, but were more active on 3' (EC 3.1.3.6)- than 5' (EC 3.1.3.5)-nucleotides. Phosphoribosyltransferase (EC 2.4.2.7,.8 and .22) and nucleoside kinase (EC 2.7.1.20) activities were detected in both forms. Nucleotide-interconverting enzymes were found to be present, with IMP dehydrogenase (EC 1.2.1.14) being the most active. Cell fractionation experiments revealed that, in the promastigote, enzyme separation within the parasite may play an important part in regulating cellular purine metabolism.     

Role of verocytotoxigenic Escherichia coli in cattle and buffalo calf diarrhoea

Abstract 273 Strains of Escherichia coli isolated from diarrhoeic and healthy control cattle and buffalo calves in Sri Lanka, were tested for Verocytotoxin (VT) and for heat-stable (ST) and heat-labile (LT) enterotoxins. VT and ST toxigenic E. coli were significantly associated with diarrhoea, accounting for 28% and 18% of diarrhoeic episodes, respectively. LT toxigenic E. coli were not significantly associated with diarrhoea.     

Mathematical analysis of the stimulus for the lateral line organ

Behavioral studies have shown that a blind fish is capable of detecting and recognizing stationary objects in its surroundings. It is proposed that the displacement of water caused by the fish as it moves is the basis for this detection capability. Alterations in the displacement of water around the fish, caused by the obstacle, act as stimuli for the lateral line organ. The question of how these stimuli acting on the skin of the fish, image the environment and what information is thus made available to the fish is the concern of this paper. The stimuli for the lateral line organ are derived mathematically. Two cases are treated: that of a fish gliding past an obstacle and that of one approaching an obstacle.     

Lineage-specific expression of c-fos and c-fms in human hematopoietic cells: Discrepancies with the in vitro differentiation of leukemia cells

In vitro differentiation studies using the bipotential human leukemia cell line, HL60, have indicated that high levels of expression of two proto-oncogenes, c-fos and c-fms, are restricted to the myelomonocytic lineage. No such expression has been detected in induced granulocytic cells. In striking contrast to these observations, we found that c-fos mRNA levels are very high in purified human granulocytes, but barely detectable in blood monocytes and tissue macrophages. Human granulocytes contain, however, relatively low levels of c-fos protein, indicating that c-fos mRNA is inefficiently translated or that the protein is rapidly degraded in these cells. In closer agreement with the in vitro results, the level of the expression of c-fms is high in purified blood monocytes and undetectable in granulocytes. We found, however, that the evolution of monocytes into tissue macrophages is accompanied by a significant decrease in c-fms expression, suggesting that the function of c-fms is restricted to specific stages of monocytic differentiation. Our observations also show that results obtained using in vitro differentiation systems have to be regarded with caution, since they may not reflect the in vivo situation.     

Serotypes of verocytotoxigenic Escherichia coli isolated from cattle and buffalo calf diarrhoea

Abstract Parasporal crystals of the recently isolated Bacillus thuringiensis var. tenebrionis are toxic for coleopteran larvae. Unlike those of other strains they are soluble either in aqueous solutions of NaBr at neutral pH or in water after titration to pH values above pH 10.0. The dissolved crystal protein readily forms crystals after removal of the salt or neutralization. The crystal protein was not found to differ much in the amino acid composition from other crystal proteins. The parasporal crystals are composed of subunits of M _r 68 000 which are not linked by disulfide bridges.