Spectrophotometric assay of immobilized tannase

A procedure for the assay of immobilized tannase with Polyacrylamide gel, collagen and Duolite-S-762 as matrices is described. It is based on the spectrophotometric determination of gallic acid formed by the enzymatic hydrolysis of tannic acid. The kinetic parameters of the enzymatic reaction have been studied and an assay procedure has been formulated. This method appears to be much more accurate than those reported earlier.     

Synthesis of lysine-containing fragments of the Proteus mirabilis O27 O-specific polysaccharide and neoglycoconjugates therefrom.

Amide-linked lysine mono- and di-uronic acid fragments of the O-specific polysaccharide from P. mirabilis O27 have been synthesised. N epsilon-Boc-L-lysine tert-butyl ester was condensed with 2-azidoethyl glycosides of glucuronic acid and beta-D-GlcpNAc-(1----3)-beta-D-GlcpA. Transformation of the products into 2-acrylamidoethyl glycosides, followed by deprotection using trifluoroacetic acid, gave the target monomers that were converted into high-molecular-weight copolymer-type neoglycoconjugates.     

Identification of a new group-specific determinant on hepatitis B surface antigen with a synthetic peptide.

In a recent study we demonstrated that a synthetic peptide representing residues 124-147 of the major protein of hepatitis B surface Ag (HBsAg) undergoes spontaneous oligomerization to reconstruct one or more conformational group-specific determinants on HBsAg. The present study was undertaken to identify and characterize the HBsAg-related antigenic determinants on this oligomeric peptide (peptide OS[124-147]). A panel of nine analogs of this peptide was generated by either deleting, substituting, or chemical side chain modification of specific amino acid residues. With HBsAg subtype-specific antisera a single "a" epitope was identified as one that includes Met133 and Lys141. In addition a "d" epitope toward the amino-terminal end of the sequence was also observed. Perturbation of certain amino acid residues was found to enhance a antigenicity and subsequent experiments indicated that maximal expression of this a antigenicity is dependent in part on accessibility of the Lys141 side chain and in part on the primary sequence. With a total of 50 human anti-HBsAg serum samples obtained from individuals vaccinated against hepatitis B, it was demonstrated that these sera recognize the Met133-Lys141-dependent a epitope as the dominant, and in many cases the only, determinant on peptide OS[124-147]. Finally, on immunization, peptide OS[124-147] elicits an anti-HBsAg response that is predominantly anti-a though a lesser contribution from an anti-d response was also obtained.     

DNA damage and repair in brain: relationship to aging.

The usefulness of conducting DNA damage and repair studies in a postmitotic tissue like brain is emphasized. We review studies that use brain as a tissue to test the validity of the DNA damage and repair hypothesis of aging. As far as the accumulation of age dependent DNA damage is concerned, the data appear to overwhelmingly support the hypothesis. However, attempts to demonstrate a decline in DNA repair capacity as a function of age are conflicting and equally divided. Possible reasons for this discrepancy are discussed. It is suggested that assessment of the repair capacity of neurons with respect to a specific type of damage in a specific gene might yield more definitive answers regarding the role of DNA repair potential in the aging process and as a longevity assurance system.     

Amide isosteres of lexitropsins: synthesis, DNA binding characteristics and sequence selectivity of thioformyldistamycin.

The synthesis and properties of an amide isostere of the antibiotic distamycin, thioformyldistamycin 3 is described. Compound 3 exists predominantly in the E conformation of the thioamide group in freshly prepared DMSO solution but is converted into the Z form, predicted by molecular mechanics to be more stable, on standing for 24 h. The coalescence temperature in DMSO is 110 degrees C by 1H-NMR. The thioformyl moiety of 3 is resistant to both peptidase action and acid treatment. Complementary strand MPE footprinting on a EcoRI/Hind III restriction fragment of pBR322 DNA demonstrated that either E or Z forms of 3 give a single set of footprints very similar to that of the parent antibiotic with strongest protection at TAAG and TATTAT with moderately strong protection at ATTT and AAAA. The strength of binding of 3 and distamycin from delta Tm measurements to either poly.d(AT) or calf thymus DNA is comparable. Molecular modeling predicted a preferred conformation for 3 wherein the C = S bond has a torsional angle of 110 degrees with the pyrrole ring. The energy difference between this conformation and the E form is less than 1 kcal/mole. In contrast the E-form has an energy 17.3 kcal/mole greater than the Z and a value of 26.3 kcal/mole was calculated for the energy barrier between the two isomers.     

Brugia malayi and Brugia pahangi: transmission blocking activity of ivermectin and brugian filarial infections in Aedes aegypti

Brugia malayi- or Brugia pahangi-infected, microfilaremic jirds (Meriones unguiculatus) were treated with ivermectin at a single dose of 200 micrograms/kg body weight, administered subcutaneously. After different time intervals, Aedes aegypti mosquitoes were fed on treated or untreated jirds. Sausage stage, L2, and L3 larvae failed to develop in mosquitoes that fed on jirds from 15 to 30 days post-treatment. After 1 month, the numbers of L3 larvae recovered from mosquitoes fed on treated B. pahangi jirds were comparable to controls. However, the number of L3's recovered from mosquitoes fed on B. malayi jirds remained significantly lower than controls, 2 and 3 months after treatment. This reduction suggests that ivermectin may be more effective in blocking transmission of B. malayi than B. pahangi. Ivermectin treatment had no effect on the mean number of circulating microfilariae in treated jirds. Therefore, mosquitoes ingested comparable numbers of microfilariae when compared to those mosquitoes fed on untreated controls. Only in the case of jirds infected with B. malayi did the circulating microfilarial counts fall 30 days after treatment. The failure of microfilariae to develop to the L3 stage in mosquitoes fed on jirds within 30 days of treatment was not due to failure of mosquitoes to ingest microfilariae. Brugia malayi microfilariae also failed to develop to L3 in mosquitoes that were allowed to feed on microfilaremic jird blood treated with ivermectin (50 ng/ml) in vitro, indicating its efficacy at low concentrations. In addition to N-acetyl glucosamine, microfilariae obtained for a period of 15 days from ivermectin-treated but not control jirds showed D-mannose, N-acetyl galactosamine, and L-fucose moieties on the surface of the sheath.(ABSTRACT TRUNCATED AT 250 WORDS)     

The cincinnati lipid research clinic family study: Familial determinants of plasma uric acid

Summary Commingling analysis of plasma uric acid levels in a random sample of 160 nuclear families supports the hypothesis that there is a mixture of three distributions. Assuming one, two, and three components in the underlying distribution, we obtained the corresponding p-values (for power transformation) as 0.059, 1.040, and 1.643, respectively. Path analysis with p=0.059 gives genetic (h ²) and cultural (c ²) heritabilities as 0.256 and 0.199, without much support for intergenerational differences, assortative mating, or maternal effects. Complex segregation analysis with p=0.059 supports multifactorial inheritance, consistent with the findings of Gulbrandsen et al. (1979) and Morton (1979) in other populations. This study also fails to support a major locus hypothesis, contrary to earlier reports.This work was supported in part by N.I.H. and N.I.M.H. Grants GM 28719, and MH 31302, and by contract NO-1-HV-2-2914L from the National Heart, Lung, and blood Institute (Lipid Research Clinic's Program), General Clinical Research Center, and the CLINFO center Grant RR-00068-19     

Cytological studies on F1 hybrids of Solanum integrifolium with S. melongena and S. melongena var. insanum

S. integrifolium (2n = 24) can easily be crossed as the pistillate parent with S. melongena (2n = 24) and S. melongena var. insanum (2n = 24). However, crosses in the other direction do not succeed. Both hybrids are vigorous. Chromosome association at diakinesis and metaphase I was studied. Chromosome associations higher than bivalents were observed in the hybrids indicating structural repatterning of chromosomes. The modal chromosome association in hybrids was twelve bivalents per PMC. This is suggestive of the retention of ancestral chromosome homeologies by the taxa concerned. Despite regular meiosis both hybrids were highly pollen-sterile (about 95%), which was attributed to segregational events of the recombined chromosomes.     

Synthesis of phosphatidylcholine under possible primitive Earth conditions

Summary Using a primitive Earth evaporating pond model, the synthesis of phosphatidylcholine was accomplished when a reaction mixture of choline chloride and disodium phosphatidate, in the presence of cyanamide and traccs of acid, was evaporated and heated at temperatures ranging from 25° to 100°C for 7 hours. Optimum yields of about 15% were obtained at 80°C. Phosphatidylcholine was identified by chromatographic, chemical and enzymatic degradation methods. On enzymatic hydrolysis with phospholipase A₂ and phospholipase C, lysophosphatidylcholine and phosphorylcholine were formed, respectively. Alkaline hydrolysis gave glycerophosphorylcholine. The synthesis of phosphatidylcholine as the major compound was accompanied by the formation of lysophosphatidylcholine in smaller amounts. Cyanamide was found to be essential for the formation of phosphatidylcholine, and only traces of HCl, of the order of that required to convert the disodium phosphatidate to free phosphatidic acid were found necessary for the synthesis. This work suggests that phosphatidylcholine, which is an essential component of most biological membranes, could have been synthesized on the primitive Earth.