Adenine deaminase of a eukaryotic animal cell,Crithidia fasciculata

Adenine deaminase (adenine aminohydrolase, EC 3.5.4.2) has been found to occur in Crithidia fasciculata with a specific activity higher than that of the same enzymes of bacteria and yeasts. It is remarkable for its stability to heat, exhibiting no appreciable loss of activity after 60 min at 55 °C. It occurs in the soluble portion of cell extracts but can be released into the suspending medium by osmotic and/or cold shock.     

Critical adjustment of cysteine and glutamine concentrations for improved clonal growth of WI-38 cells

Summary Clonal growth of WI-38 cells with a plating efficiency of 45% has been achieved in a synthetic nutrient mixture (MCDB 102) supplemented with either whole or dialyzed fetal bovine serum. For optimum growth, the concentration of cysteine in the medium must be adjusted precisely. Deviation by a factor of three in either direction from the optimum concentration (9.0×10⁻⁵M) eliminates essentially all clonal growth. A high concentration of glutamine (2.5×10⁻³M) is also needed for, optimum clonal growth. Presented in preliminary form at the 26th Annual Meeting of the Tissue Culture Association, June 4, 1975. This work was supported by Grant No. HD-08181 from the National Institute of Child Health and Human Developement, Grant No. AG-00310 from the National Institute on Aging, and by Contract No. 223-74-1156 from the Bureau of Biologics, Food and Drug Administration.     

Fatty acid positional distribution in phospholipids of a psychrophilic bacterium during changes in growth temperature

The major phospholipids of the psychrophilic bacteriumMicrococcus cryophilus, phosphatidylethanolamine and phosphatidylglycerol, have similar fatty acid compositions, comprising almost entirely palmitoleic and oleic acids. We show that there is a preference for the longer chains in thesn-1 position of both phosphatidylethanolamine and phosphatidylglycerol, both during isothermal growth and after temperature shifts, despite the fact that the overall phospholipid C18/C16 acyl chain ratio decreases with a lowering of growth temperature. Although it has been shown using model systems that the isomeric configurationsn-1-long,sn-2-short lowers lipid melting temperature, this paper reports the first clear-cut demonstration of such an isomeric preference in a natural system. We discuss how this acyl chain configuration contributes to membrane fluidity inM. cryophilus, in terms of adaptation to its psychrophilic habitat.     

Inactivation of Glutamine Synthetase by Ammonia Shock in the Gram-Positive Bacterium Streptomyces cattleya

In cultures of the gram-positive bacterium Streptomyces cattleya, a rapid inactivation of glutamine synthetase was seen after ammonia shock. pH activity curves for ammonia-shocked and control cultures are shown. A peak of glutamine synthetase activity was seen during fermentation for production of the antibiotic thienamycin.     

Fertilization acid of sea urchin eggs: Evidence that it is H+, not CO2

The report of R. J. Gillies, M. P. Rosenberg, and D. W. Deamer (1981, J. Cell. Phys., 108, 115–122) that sea urchin fertilization acid is anaerobically produced CO₂, was reinvestigated by inseminating Strongylocentrotus purpuratus eggs in HCO^?₃-free seawater, then bubbling the seawater with N₂ to remove volatile acid. Fertilization acid production occurred in HCO^?₃-free seawater and with N₂-bubbling, the pH rose 0.28 ± 0.08 unit, significantly less than the rise of 0.63 ± 0.14 unit during N₂-bubbling of HCO^?₃-free seawater that had been acidified with CO₂ and similar to the rise of 0.18 ± 0.07 unit when acidification was with HCl. We conclude that most, if not all, of the sea urchin fertilization acid is nonvolatile and thus is not CO₂; since it is not a weak acid, it must be H⁺.     

Cerebral Metabolic Effects of Monoamine Oxidase Inhibition in Normal and 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Acutely Treated Monkeys

The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induces dopaminergic cell death in the substantia nigra pars compacta (SNpc) and clinical parkinsonism in humans and experimental animals. Pretreatment with monoamine oxidase inhibitors prevents this cell death and associated parkinsonism by blocking the oxidation of MPTP to a toxic intermediate. The 2-deoxyglucose method was used to study the acute effects of MPTP in the monkey brain and the effects of monoamine oxidase inhibition on local cerebral glucose utilization in both normal and MPTP-treated monkeys. MPTP administration alone caused a major increase in glucose utilization in the SNpc and smaller increases in some subnuclei within the ventral tegmental area in which eventual dopaminergic cell loss also occurs. Pretreatment with pargyline abolished these metabolic increases, a finding suggesting both that the oxidized product of MPTP generates the metabolic increases and that the increased glucose consumption may contribute to cell toxicity. On the other hand, in most cortical, thalamic, striatal, brainstem, and cerebellar areas MPTP alone caused reductions in glucose utilization, and pargyline failed to prevent these effects. Pargyline alone depressed metabolism in the locus coeruleus and a few other monoaminergic structures.     

Bovine trophoblastic cell vesicle attachment to polarized endometrial epithelial cells in vitro

Summary Interactions between bovine trophoblastic cell vesicles and bovine endometrial epithelial cells were investigated by light and electron microscopy and lectin histochemistry in a cell culture model of early blastocyst attachment. Primary lines of bovine endometrial epithelial cells were polarized by subculturing on substrata and maintaining cultures at the air-medium interface. Trophoblastic cell vesicles were obtained from elongated Day 14 blastocysts. In co-cultures, trophoblastic cell vesicles adhered to endometrial epithelial cells through microvillus interdigitation and formation of primitive membrane junctional complexes. After 3 d in co-culture, a multilayered cellular plaque formed at the trophoblastic cell-endometrial epithelial cell interface. The type of cells contributing to this local proliferative response could not be identified specifically as trophoblastic or endometrial cells, and areas of membrane fusion between cells were noted. Ultrastructural features of vesicle adhesion in cultures were similar to features of conceptus attachment in vivo. Lectins bound to apical membranes of trophoblastic cells and endometrial epithelial cells in all locations except contact sites between vesicles and endometrial cells. These findings suggest that local cellular proliferation and membrane fusion between trophoblastic and endometrial epithelial cells may be early events in conceptus implantation in the cow and these events can be reproduced in culture. This work was supported by a grant from U.S. Department of Agriculture Animal Health and Disease Program, Washington, DC.     

Conformational changes of recombinant human granulocyte-colony stimulating factor induced bypH and guanidine hydrochloride

Fluorescence and circular dichroism were used to follow thepH-dependent conformational changes of granulocyte colony stimulating factor (G-CSF). Tryptophan fluorescence of the spectra monitored at 344 nm, or after deconvolution of the emission spectra, at 345 nm, showed a decrease in intensity on going frompH 7 to 4, with a midtransitionpH of 5.8. On the other hand, tyrosine fluorescence measured either by the ratio of intensity at 308 nm to that at 344 nm, or by the fluorescence intensity at 303 nm after deconvolution of the spectra, increased in intensity as thepH was changed from 6 to 2.5, with a midtransitionpH of 4.5. Near UV circular dichroic spectra also showed changes betweenpH 7.5 and 4.5, which correlated with the transition monitored by the tryptophan fluorescence. The guanidine hydrochloride-induced conformational changes of G-CSF at fivepH values from 2.5 to 7.5 were also studied. Circular dichroic and fluorescence spectra revealed minor conformational changes by the addition of 1 or 2 M guanidine HCl at allpH values examined, while the major conformational transition occurred between 2 and 4 M guanidine hydrochloride. The secondary structure of the protein was most stable betweenpH 3.3 and 4.5. The guanidine HCl-induced denaturation of G-CSF involved more than a two-state transition, with detectable intermediate(s) present, and the structure of the intermediate(s) appeared to depend on thepH used. These results are consistent with thepH dependence of the structure described above, and demonstrate the complex conformational properties of G-CSF.     

New records of within-group infanticide and cannibalism in wild chimpanzees

Two cases of within-group infanticide and cannibalism were observed among the M Group chimpanzees of the Mahale Mountains, Tanzania. In both cases, victimized infants were male, 5 – 6 months of age, and in good health when killed. Four to five years have passed since the mothers of the victims immigrated into M Group as nulliparous immigrants. In one case the 2nd-ranking male was observed to detach the infant from the mother's belly. Both infants were finally killed by the alpha male after several adult males scrambled for the bodies. There was no evidence that the mothers had mated with males other than those of M Group. Nor was there evidence that the mothers had restrictive mating relationships with some of the M Group adult males. What little evidence is available shows that the mothers had mated mostly with adolescent and other immature males during their conception cycles. However, at least in one case, the mother began to mate more with adult males rather than with immature males after the infanticide. It is proposed that the function of within-group male infanticide can be explained by the male-male competition hypothesis developed for hanuman langurs and other nonhuman primates.