The identification of unbranched enzyme kinetic mechanisms by computer analysis of qualitative information: Logic of the method

The paper describes the logic of a computer method for identifying unbranched enzyme kinetic mechanisms on the basis of observed initial velocity and product inhibition patterns (Cleland, 1963). The method establishes initially an acceptable order of reactant addition and release, proceeds to list all the mechanisms consistent with that order and the data, and finally determines which of these can also explain data which require either non-competitive or dead-end inhibition.     

Seasonal changes in plasma testosterone and ejaculatory capacity in squirrel monkeys (Saimiri sciureus)

Seasonal changes in body weight, plasma testosterone and ejaculatory capacity were observed in five intact and two testosterone-implanted castrated squirrel monkeys for a total of 13 months. Electroejaculation was employed for obtaining data concerning ejaculation. In the intact animals, there were significant increases in body weight, ejaculate volume and plasma testosterone during the breeding season. With the exception of one animal, there was also a decrease in ejaculation latency during the season. Seasonal differences in the sperm count and sperm motility were not observed. Testosterone-implanted castrates showed changes in both ejaculate volume and ejaculation latency similar to those seen in intact monkeys during the breeding season. The body weight and plasma testosterone of the castrates remained quite constant throughout the year. Supported by NIH grants HD 00778, MH 21178 and MH 23645.     

Studies of ribosomal proteins of yeast species and their hybrids gel electrophoresis and immunochemical cross-reactions

Summary The cytoplasmic ribosomal proteins (r-proteins) of seventeen yeast species of the genera Saccharomyces and Kluyveromyces were analyzed by one-dimensional gradient polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. The electrophoretic patterns of cytoplasmic r-proteins from different species display extensive differences in both the 40S and the 60S subunit. Relatedness of species suggested by r-protein patterns correlates well with that based on DNA/DNA homology (Bicknell and Douglas 1970). Immunochemical cross-reactions and antibiotic susceptibility tests were also used to compare different species.Analyses of r-proteins from two different interspecific hybrids showed that their ribosomes were hybrid, containing r-proteins from both parents. These findings are discussed in relation to the evolution of yeast species and the regulation of expression of r-proteins in cucaryotes.     

A new species of Eugenia (Myrtaceae) from Venezuela

Eugenia mcvaughii is described from a forest remnant of the Jardin Botánico of Caracas, Venezuela. Found also in the hills adjacent to Caracas, it may be considered as an endemic species of the Interior Coastal Cordillera.     

Trypanosoma cruzi: allopurinol in the treatment of mice with experimental acute Chagas disease

The therapeutic effect of allopurinol was studied in an experimental Trypanosoma cruzi infection (Chagas disease) in outbred IVIC-NMRI and inbred C57B1/6J mice intraperitoneally inoculated with the parasites 2–6 days before drug treatment. Allopurinol protected against T. cruzi infection. This effect was evidenced by highly significant reductions in both parasitemias and mortality rates and increased survival time in allopurinol-treated animals compared with untreated infected mice. Allopurinol protected effectively when administered in 10 daily doses of 32–64 mg/kg body wt/day injected intraperitoneally. Using direct methods, parasitemia remained undetectable for at least 310 days. An indirect method, subinoculation to susceptible mice, showed a few circulating trypanosomes which decreased greatly in number after a second schedule of allopurinol treatment; finally no trypanosomes were detectable 275 days after treatment initiation. Allopurinol also induced a strong trypanostatic effect when tested in vitro on five different Trypanosoma cruzi strains (optimal inhibitory concentration: 3 μg/ml). These results suggest that allopurinol protects mice with acute Chagas infection by a direct trypanostatic effect. The low toxicity of this drug suggests its use in more chronic experimental Chagas infections.     

New species from the summit of Cerro Guaiquinima,Venezuela

Three new species,Lubaria szczerbanii (Rutaceae),Croton guaiquinimae (Euphorbiaceae), andStomatochaeta colveei (Asteraceae), are described from the summit of Cerro Guaiquinima, Venezuela. The generic affinities betweenLubaria andMyllanthus, and betweenStomatochaeta andStenopadus are discussed.     

Trypanosoma cruzi: defined medium for continuous cultivation of virulent parasites.

A liquid medium was developed for the continuous cultivation of Trypanosoma cruzi. Among the several highly purified macromolecules tested only bovine liver catalase, horseradish peroxidase, lactoperoxidase, and bovine hemoglobin supported the continuous growth, at high yield, of mice-virulent Trypanosoma cruzi; other hemoproteins were inactive. Bovine liver catalase showed optimal Trypanosoma cruzi growth-promoting activity, parasites reaching 20 × 10⁶ parasites/ml (95% epimastigotes) at about 10 days in most of the 45 subpassages to date. Furthermore, this protein in the incubation medium provided all the amino acid requirements of actively growing parasites, thus eliminating the need for exogeneous free amino acids. Additional experiments revealed that the hemoprotein's growth-promoting activity was independent of any enzymatic activity and that reconstituting the exact protein composition by means of exogeneous amino acids did not support parasite multiplication, suggesting the importance of the primary structure of the active proteins for growth-promoting activity. These active macromolecules supported the multiplication of five different strains of Trypanosoma cruzi, but did not support Leishmania brasiliensis or Leishmania mexicana proliferation, suggesting species specificity.     

New taxa from the Avila and Naiguatá Mountains,Venezuela

Ten new species (Billbergia manarae, Croizatia naiguatensis, Croton huberi, Rinorea oraria, Anaectocalyx manarae, Linociera avilensis, Cynanchum beatricis, C. manarae, Matelea costanensis, and M. manarae) are described in preparation for the forthcoming Flora of Avila and Naiguatá.     

Binding of microtubule proteins to DNA: specificity of the interaction.

Tubulin is detected among the DNA-binding proteins when an extract from fibroblasts is chromatographed on DNA-cellulose. Further purification of the colchicine-binding activity shows that purified tubulin from fibroblasts does not bind to DNA. Depolymerized brain microtubule proteins show a high affinity for DNA. The fraction bound is composed of tubulin and microtubule-associated proteins. Experiments with fractionated microtubule proteins indicate that tubulin-free microtubule associated proteins bind to DNA, while tubulin free of microtubule-associated proteins does not. Microtubule-associated proteins bind better to eukaryotic than to phage DNA suggesting a specificity of the interaction.