DNA strand scission and ADP-ribosyltransferase activity during murine erythroleukemia cell differentiation

The accumulation of DNA strand breaks and activation of ADP-ribosyltransferase (ADPRT) have recently been associated with cellular differentiation. Murine erythroleukemia (MEL) cells undergo erythropoietic differentiation when exposed to dimethyl sulfoxide (Me2SO) and several studies have suggested that DNA strand scission induced by this agent is a prerequisite for expression of the differentiated phenotype. Me2SO induction of MEL cells has also been associated with increases in ADPRT activity in one study, but not in another. We have monitored the effects of Me2SO on DNA strand breaks in preformed and replicating MEL cell DNA. The results clearly demonstrate that DNA fragmentation is not detectable during Me2SO induction of MEL differentiation, even in the presence of 3-aminobenzamide, an inhibitor of ADPRT. Further, these results are consistent with an absence of detectable changes in both endogenous and total potential ADPRT activity during Me2SO-induced MEL differentiation. These findings would argue against Me2SO induction of DNA strand scission and ADPRT in MEL cells undergoing differentiation.     

Stabilization of a histidine-producing strain of Serratia marcescens.

A decrease in histidine productivity was observed during subculture of a histidine-producing strain of Serratia marcescens. The decrease was accompanied by an increase in the number of wild-type revertants. Adenine accelerated the growth of producing strain HT-2892 to nearly equal that of revertants, and histidine production was stable because the depletion of ATP in strain HT-2892 was restored by adenine. To increase the intracellular ATP content, mutants resistant to 6-methylpurine, an antagonist of adenine, were isolated from strain HT-2892. 6-Methylpurine-resistant mutant MPr90 grew more rapidly than the parent producing strain and produced L-histidine stably, even when it was subjected to subculture in medium without adenine. ATP depletion was restored in strain MPr90, probably owing to the derepression of adenylosuccinate synthetase in AMP biosynthesis.     

Comparative enzyme histochemical study on the visceral yolk sac endoderm in the rat in vivo and in vitro

Summary Histochemical study of the visceral yolk-sac endoderm of the rat was performed in vitro (whole-embryo culture for 24, 48 and 72 h explanted at 9.5 days of gestation) and in vivo (10.5, 11.5 and 12.5 days of gestation) in order to compare the distribution and activity of various enzymes involved in the digestion and energy metabolism in both systems. It was shown that, both in vitro and in vivo -glytamyltransferase and dipeptidylpeptidase IV are demonstrable in the apical cell membranes (membranebound hydrolases), while acid phosphatase, dipeptidylpeptidases I, II and acid -galactosidase are concentrated in the supranuclear vacuoles (lysosomal hydrolases), and cytoplasmic lactate dehydrogenase and mitochondrial enzymes (succinate dehydrogenase, NAD-dependent isocitrate dehydrogenase, cytochrom oxidase) are localized in the whole cytoplasm and mainly in the apical cytoplasm, respectively, of the visceral yolk-sac epithelium. In vivo, the activity of all enzymes increased until 12.5 days, but in vitro, this activity increased only until 48 h after the start of culture (corresponding to 11.5 days in vivo). Comparison of the yolk sacs at 10.5 and 11.5 days in vivo with those after 24 and 48 h in vitro showed that the activities of all the investigated enzymes were almost identical. Yolk sacs which were cultured for 72 h showed lower activities of lysosomal and mitochondrial enzymes than those at 12.5 days in vivo. It is concluded that the digestive function and energy metabolism of the visceral yolk-sac epithelium are almost identical in vitro and in vivo at 10.5 and 11.5 days.Supported by the Deutsche Forschungsgemeinschaft 541/1-1)     

Plasma growth hormone and thyrotropin responses to thyrotropin releasing hormone in freely behaving and urethane anesthetized rats.

Plasma GH and TSH responses to thyrotropin releasing hormone (TRH) were examined in freely behaving and urethane anesthetized rats. The i.v. administration of TRH (200ng/100g b.wt.) resulted in consistent elevations of plasma GH only in urethane anesthetized rats, while significant elevations of plasma TSH were similarly observed in both conditions. Results suggest that urethane influences plasma GH responses to TRH.     

The synthesis and chromatography of peptide nitriles

Di- and tripeptide nitriles, glycylaminoacetonitrile (Gly-AAN), diglycylaminoacetonitrile (Gly-Gly-AAN), alanyl--aminopropionitrile (Ala-APN), and dialanyl--aminopropionitrile (Ala-Ala-APN) were synthesized first.These peptide nitriles and related peptides and peptide amides were analyzed by means of ion-exchange chromatography. The every two diastereomers of dialanine, dialanine amide, and Ala-APN were separated into two peaks by using a pH 3.25 buffer as an eluent. The four isomers of trialanine, trialanine amide, and Ala-Ala-APN gave four, two, and one peak, respectively under the same conditions.The trimethylsilyl derivatives of alanyl peptides and related compounds were analyzed by means of gas chromatography combined with mass-spectrometry. The parent (M⁺ and/or M⁺-15) and other mass numbers observed in their mass-spectra supported the introduction of various numbers of trimethylsilyl groups.     

Serotonin in neuroblastoma × glioma NG108-15 hybrid cells

Mouse neuroblastoma × rat glioma NG108-15 hybrid cells contain a considerable amount of serotonin, and possess small but significant tryptophan hydroxylase activity. The results suggest that NG108-15 hybrid cells are serotonergic, in addition to the known cholinergic property.     

The complete nucleotide sequence of a 23-S rRNA gene from tobacco chloroplasts

The nucleotide sequence of a tobacco chloroplast 23-S rRNA gene, including the spacer between it and the 4.5-S rRNA gene, has been determined. The 23-S rRNA coding region is 2804-base-pairs long. A comparison with the 23-S rRNA sequence of Escherichia coli reveals strong homology and further shows a similarity between the chloroplast 4.5-S rRNA and the 3'-terminal region of E. coli 23-S rRNA. However, the 101-base-pair spacer sequence between the 23-S and 4.5-S rRNA genes has little homology with E. coli 23-S rRNA.     

Purification, characterization, and amino acid sequences of pepsinogens and pepsins from the esophageal mucosa of bullfrog (Rana catesbeiana)

Two pepsinogens (pepsinogens 1 and 2) were purified from the esophageal mucosa of the bullfrog (Rana catesbeiana), and their molecular weights were determined to be 40,100 and 39,200, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The NH2-terminal 70-residue sequences of both pepsinogens are the same, including the 36-residue activation segment. Furthermore, a cDNA clone encoding frog pepsinogen was obtained and sequenced, which permitted deduction of the complete amino acid sequence (368 residues) of one of the pepsinogen isozymogens. The calculated molecular weight of the protein (40,034) coincided well with the values obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results are incompatible with the previous report (Shugerman R. P., Hirschowitz, B. I., Bhown, A. S., Schrohenloher, R. E., and Spenney, J. G. (1982) J. Biol. Chem. 257, 795-798) that the major pepsinogen isolated from the bullfrog esophageal gland is a unique "mini" pepsinogen with a molecular weight of approximately 32,000-34,000. The two pepsinogens were immunologically indistinguishable from each other and related to human pepsinogen C. The deduced amino acid sequence was also more homologous with those of pepsinogens C than those of pepsinogens A and prochymosin. These results indicate that the frog pepsinogens belong to the pepsinogen C group. They were both glycoproteins, and therefore, this is the first finding of carbohydrate-containing pepsinogens C. Both pepsinogens were activated to pepsins in the same manner by an apparent one-step mechanism. The resulting pepsins were enzymatically indistinguishable from each other, and their properties resembled those of tuna pepsins.     

Anti-pituitary antibodies in patients with hypopituitarism and their families: longitudinal observation.

In an attempt to investigate the clinical significance of anti-pituitary antibodies in patients with hypopituitarism, anti-pituitary antibody in plasma was examined in 10 such patients (7 cases of isolated ACTH deficiency, 1 of partial hypopituitarism, and 2 of Sheehan's syndrome), on two or three occasions with an interval of more than 6 months (longitudinal study). In a total of 16 relatives of these 4 patients (2 cases of Sheehan's syndrome, one in each of partial hypopituitarism and isolated ACTH deficiency) and one patient not involved in the longitudinal study, anti-pituitary antibodies were also examined (family study). Anti-pituitary antibodies reacting with rat pituitary cytoplasmic antigens (pituitary cell antibodies: PCA) and pituitary cell surface antibodies (PCSA) reacting with GH3 cells and/or AtT-20 cells were measured with indirect immunofluorescence. The longitudinal study revealed the disappearance of antibodies in 3 patients, 2 PCA positive and one both PCA and PCSA positive. In 3 patients, altered antibody titers or a newly appearing antibody were found during the follow-up period. In 4 patients, the pituitary antibodies were negative during the follow-up periods. Of 16 family members studied, positive PCA was found in 3 members (2 in the families of patients with PCA positive Sheehan's syndrome, and 1 in the family of the patients with PCA positive partial hypopituitarism). Positive PCSA was found in 4 members (one in each of families of patients with partial hypopituitarism and isolated ACTH deficiency and of two cases of Sheehan's syndrome), and weakly positive PCSA was found in one family member of a patients with PCA positive Sheehan's syndrome.(ABSTRACT TRUNCATED AT 250 WORDS)