Serum-dependent depression of alkaline phosphatase in HeLa cells

Reduction in alkaline phosphatase activity was observed when HeLa S3 cells were grown in Puck's medium containing high concentrations of human serum. This effect was not seen with the enzyme of Chang liver 8A cells. The induction of increased alkaline phosphatase in HeLa S3 by prednisolone or by osmolality changes was not prevented by serum. The concentration of serum in the culture medium had no influence on acid phosphatase activity.     

Hormonal Regulation of Cell Elongation in the Hypocotyl of Rootless Soybean: An Evaluation of the Role of DNA Synthesis

A method was developed where soybean seedlings were grown without roots to study the influence of hormones of root origin on shoot growth. Excision of the root resulted in inhibition of apical section growth and DNA synthesis and inhibited elongating section growth. A synthetic cytokinin restored DNA synthesis in the apical section, but did not influence growth in either the apical or elongating sections. Low concentrations of gibberellin with the cytokinin restored growth in the apical section. Gibberellin alone was sufficient to restore growth in the elongating section.An inhibitor of DNA synthesis, 5-fluorodeoxyuridine, inhibited the increase in apical section DNA without inhibiting control or gibberellin-induced growth in the elongating section. Experiments with (14)C-thymidine resulted in no DNA labeling differences in the elongating section under conditions where gibberellin-induced elongation varied from 50% to 73% above controls. It was concluded that gibberellin-induced elongation in soybean hypocotyl occurred in the absence of DNA synthesis. Gibberellin does stimulate DNA synthesis in the apical tissue apart from its effect on cell elongation.Excised soybean hypocotyl elongated maximally at 10(-6)m auxin. At higher auxin concentrations, fresh weight and ethylene production increased, but elongation was reduced. Addition of GA to the higher auxin concentrations resulted in a 50% inhibition in auxin-induced ethylene production and resumption in maximal elongation. Added ethylene inhibited elongation 30% at 2 mul/l. Addition of up to 100 mul/l ethylene did not inhibit elongation with GA present in the incubation medium. Thus GA may counteract ehtylene inhibition of cell elongation in addition to inhibiting ethylene production in auxin-treated tissues.     

Influence of Calcium and Magnesium on Manganese Absorption

Mutual effects between Mn, Ca, and Mg were studied during steady-state absorption experiments with excised barley roots. Calcium appeared to enhance the rate of Mn absorption; whereas, Mg had a highly depressive effect. The combination of both Ca and Mg was even more inhibitory to Mn absorption than Mg alone. Manganese had no effect on the usual negligible Ca absorption by this tissue, but effectively inhibited the absorption of Mg. Although divalent cation absorption from the Ca-Mg-Mn system was essentially nil, K absorption was greatly stimulated in the presence of these cations.These mutual effects and others reported in the literature are explained by the hypothesis that selectivity in ion absorption results from cation-induced conformational changes in the structure of the carrier molecule.     

Fine structural study of gas secretion in the physoclistous swim bladder of Fundulus heteroclitus and Gadus callarias and in the euphysoclistous swim bladder of Opsanus tau

Summary The Haldane-Koch-Scholander-Kuhn-Steen theory of salting out countercurrent multiplication effect of the rete mirabile now accounts for release of most gases in the fish swim bladder. Evidence presented here indicates that final release is by microbubbles from the secretory epithelium. There is only one specific cell type in a highly vascularized epithelium. It is characterized by complex folds in the paravascular zone and by gas forming bodies which seem to form from plentiful Golgi material. The bodies are formed with dark amorphous matrix that becomes patterned (tubular or lamellar), finally froths and then is released to the gas surface. Residual material may form myelin-like layers on the lumenal surface. Active cells are also characterized by surface villi and subsurface, parallel cisternal spaces. Gas may be formed by cells not touching the gas surface and released through intercellular spaces. There are discontinuous desmosomes (maculae adhaerentes) near the gas surface and there are no tight junctions (zonulae occludentes). Gas release as bubbles would explain Wittenbergs observations that the gases found in swim bladders have ratios more closely related to their solubility coefficients in water than to ambient partial pressures. A surfactant may be present to lower the surface tension of the microbubbles. The carrier in the cytoplasm would have to be an iron-protein (or perhaps peroxidase) compound capable of binding molecular oxygen.Supported by grant-in-aid from the national Science Foundation (GB-676) and from the USPHS (General Medical Sciences Institute, GM-06836).     

B12 Coenzyme-dependent Ribonucleotide Reductase in Rhizobium Species and the Effects of Cobalt Deficiency on the Activity of the Enzyme

This investigation revealed that the ribonucleotide reductases in extracts of Rhizobium leguminosarum, R. trifolii, R. phaseoli, R. japonicum, and R. meliloti 3DOal (ineffective in nitrogen fixation) are dependent upon B(12) coenzyme for activity. Rhizobium and certain Lactobacillus species are the only two groups of organisms known to contain B(12) coenzyme-dependent ribonucleotide reductases. Extracts of cobalt-deficient R. meliloti cells assayed in the presence of optimum B(12) coenzyme showed a 5- to 10-fold greater ribonucleotide reductase activity than comparable extracts from cells grown on a complete medium. Furthermore, cobalt-deficient cells were abnormally elongated and contained reduced contents of deoxyribonucleic acid. The addition of purified deoxyribonucleosides to cobalt-deficient cultures of R. meliloti failed to alleviate deficiency symptoms.     

Gene-Enzyme Relationships in Histidine Biosynthesis in Bacillus subtilis

Mutants for 9 of the 10 steps in histidine biosynthesis have been isolated and identified by enzyme assay. Each locus has been mapped in relation to the aro cluster and to other histidine loci by deoxyribonucleic acid-mediated transformation. The genes which code for enzymes 3, 6, and 8 of the pathway are linked to the aro cluster. A major histidine linkage group is composed of the genes which specify enzymes 1, 2, 5, 7, and 10. The locus which codes for step 9 of the pathway is unlinked to any other identified his loci. The major histidine cluster is loosely linked to cysB and is unlinked to any of the loci concerned with aromatic amino acid biosynthesis.     

Deoxyribonucleic Acid Synthesis During Microcyst Germination in Myxococcus xanthus

Deoxyribonucleic acid (DNA) synthesis was measured during microcyst germination in Myxococcus xanthus by radioactive thymidine incorporation, autoradiography, and chemical analysis. Microcysts contained an average of 6.6 conserved units of DNA, corresponding to 3 to 4 chromosomes per cell. Correlation of the DNA content and chromosome number of microcysts indicated that the molecular weight of the nonreplicating M. xanthus chromosome is 4.9 x 10(9) daltons. DNA synthesis was initiated 3.5 to 4 hr after induction of germination. From 4 to 6 hr, the rate of synthesis was constant and the accumulation was linear. After a lag period (6 to 6.5 hr), the rate of DNA synthesis increased, reaching a second plateau at 9 hr. From 9 to 11 hr, the rate was again constant and the accumulation was linear. Cellular division during germination showed an unusual kind of synchrony. A model is presented that accounts for chromosomal replication and cell division during microcyst germination.     

Distribution of β-thalassemia trait and erythrocyte glucose-6-phosphate dehydrogenase deficiency in the Markham river valley of New Guinea

Ten villages in or near the Markham Valley, northeastern New Guinea, have provided a sample of 476 males for the ascertainment of glucose-6-phosphate dehydrogenase (G6PD) deficiency and 810 individuals of both sexes for the investigation of the β-thalassemia trait. An extreme heterogeneity was found in the prevalence of both traits when the villages were analyzed separately (from 1.5% to 18.2% demonstrating G6PD deficiency and from 0 to 22.8% evincing β-thalassemia trait). The latter result counters an earlier report from the same region that β-thalassemia trait frequencies correlated negatively with altitude and (presumably) positively with endemicity of malaria. The present findings, based on a more representative sample, do not necessarily invalidate the relationship between malaria and β-thalassemia trait suggested by earlier studies, but they do make clear that correlation in New Guinea, at least, may be complicated by random genetic drift, although other possibilities are discussed. The heterogeneous distribution of G6PD deficiency in the present study confirms similar earlier findings. The importance of sample provenience and anthropological data in the interpretation of differences in the distribution of genetically determined traits is stressed.     

MICROSPECTROPHOTOMETRIC ANALYSIS OF NUCLEAR DNA IN CHARA ZEYLANICA

Microspectrophotometric analysis of the DNA content of nuclei in various parts of Chara zeylanica Willd. revealed that the amount of DNA in the nucleus of an internodal cell equals twice the amount of DNA in the nucleus of a sperm, while the half-anaphase stage of the same nodal cells contains the same amount of DNA as the nuclei of the male gametes. The DNA content of the nuclei of internodal cells may rise as much as 50 times higher than that of the gametes. However, in the oldest (most basal) internodal cells, the DNA content of the minute nuclei falls again to the basic (1 C) amount. Measurements of sister nuclei derived by amitosis indicated that both nuclei have equal amounts of DNA; this was interpreted as further evidence that amitosis is not a disorganized process or manifestation of degeneration. The bearing of these analyses on the question of the site of meiosis in these plants is discussed.