Antimetabolic activities of 2-fluoro-L-histidine.

2-Fluoro-L-Histidine inhibits protein synthesis in various cell cultures, as measured by ³H-leucine incorporation. This histidine analog also inhibits the cytopathogenicity of a number of RNA and DNA viruses in primary and continuous cell cultures; it blocks the transformation of normal mouse (MO) cells by murine sarcoma virus, and partially suppresses the release of murine leukemia virus by a continuously infected mouse cell line (JLSV5). In human skin fibroblasts, it reduces the interferon-inducing capacity of poly(I)·poly(C). Inhibition of cell protein synthesis may be the common cause of the various effects. 4-Fluoro-L-histidine is essentially inert in all of the test systems examined.     

Na+,K+-ATPase: on the nature of the "cross-linked" subunits obtained in the presence of o-phenanthroline and cupric ion.

In previous studies on the quaternary structure of Na⁺,K⁺-ATPase, cupric-phenanthroline complex (CP) has been used for the cross-linking of the enzyme subunits. Here we show that the same products obtained in the presence of CP (α,α-dimer, α,β-dimer, and products of higher molecular weight) are also obtained when the enzyme is exposed to Cu²⁺ without o-phenanthroline. The α,β-dimer (but not the α,α-dimer) is dissociated in the presence of EDTA; indicating that this product is not the result of the covalent cross-linking of the subunits through a disulfide bond. The nature of the α,α-dimer remains to be determined. The findings suggest that the results of “cross-linking” experiments with CP should be interpreted with caution until the products are more clearly identified.     

A spectrofluorimetric investigation of calf thymus DNA modified by BP diolepoxide and 1-pyrenyloxirane

7β,8α-Dihydroxy-9α,10α-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BP diolepoxide, $\text{1}$) and 1-pyrenyloxirane ($\text{2}$) bind chemically to calf thymus DNA. The fluorescence efficiency of pyrenyl groups in mutagen modified DNA varies appreciably with its conformation and decreases in the order: pyrenees, modified denatured DNA and modified native DNA. A particularly interesting observation is that the fluorescence efficiency of mutagen modified DNA intensifies substantially upon denaturation. Our results suggest that the pyrenyl groups in mutagen modified DNA are intercalated between the base pairs of DNA. Since both $\text{1}$ and $\text{2}$ are powerful frame-shifting mutagens for S. typhimurium TA-98, the intercalative covalent binding of these compounds to DNA may provide a molecular basis for their mutagenic activity.     

Biosynthesis of a ubiouitin-related peptide in rat brain and in human and mouse pituitary tumors

A biosynthetic labeled peptide structurally related to the thymic peptide ubiquitin was first identified fortuitously in bovine pars intermedia cells in regard to its partial NH₂ terminal amino acid sequence (Met 1, Leu 8, 15 and Lys 6, 11, 27, 29, 33) after a protein segment data bank search. A peptide with the same behavior on carboxymethylcellulose chromatography and polyacrylamide gel electrophoresis has been purified after labeling experiments in two areas of rat brain, hypothalamus and striatum, and in a mouse and a human ACTH-secreting pituitary tumors. It represents about 1 to 10% of the total labeled proteins in the various experiments. Its identity with the above mentioned bovine pituitary peptide was confirmed by microsequence analysis with respect to Met 1, Lys 6, 11 in hypothalmus, Met 1 in striatum, and Lys 6, 11, 27, 29, 33 in the two pituitary tumors. The availability of standard purified ubiquitin allowed us to show that labeled and cold peptides have the same electrophoretic mobility and elution volume on Sephadex G-50 chromatography this further confirms their identity. Possible interests of such a biosynthetic characterization of a ubiquitin-related peptide are discussed, particularly in view of the structural relationship of ubiquitin to the non histone component of nuclear protein A-24, and as a test of tissue viability and biosynthetic efficiency in our in vitro biosynthetic systems.     

Boophilus microplus: characterization of larval proteases.

Cercariae of Schistosoma mansoni were treated with undecenyl-pseudothiourea. After centrifugation, they agglutinated into a mass. Resuspended in water, they remained immobilized. When injected sub-cutaneously into mice, they produced bisexual infections. The immobilizing drug effect, together with a reduced worm recovery rate, are time and concentration dependent. The cercariae become avirulent (99.8%) only when the flame cell is affected. Immobilizing and “cercaricidal” effects are not necessarily related properties; the latter can be determined only by in vivo tests of infectivity. No protection against reinfection was noticed in mice injected with immobilized cercariae of reduced virulence. The immobilized cercariae produced infections with a 0.7% worm recovery rate by percutaneous exposure, compared to 2.2% by subcutaneous injection. Normal cercariae produced infections with average recovery rates of 11.1% subcutaneously and 45% percutaneously.     

The occurrence of the second moult of Ascaris lumbricoides and Ascaris suum

Maung M. 1978. The occurrence of the second moult of Ascaris lumbricoides and Ascaris suum. International Journal for Parasitology 8: 371–378. Eggs of Ascaris lumbricoides and A. suum were cultured at 28°C and observed daily. Larvae were released by pressure, by artificial hatching with CO₂, and by natural hatching after infection of laboratory mice. The early stages of development in the egg were observed to comprise two moults, one occurring immediately after the other. Both moults were initiated within the egg, but the time of completion of the second moult varied considerably, and in some instances was not completed until the larvae reached the liver of experimentally infected animals.     

Relationship between acute and chronic exposures in mutagenicity studies in mice

The relationship between acute and chronic exposures in mutagenicity studies on mammals still lacks experimental data that might permit the decision whether or not the long-term exposures are of significance in mutagenicity testing.Fractional application of TEPA, THIOTEPA, EMS cyclophosphamide and sodium arsenite was made in experiments with mice, by using the dominant-lethal test and cytogenetic analysis of bone marrow. In most experiments the repeated application yielded the same or higher genetic injury than the same total dose at a single application. Negative results are discussed in relation to the threshold dose and the different sensitivity of the germ-cell stage.Possible interaction of mutagens was also studied by analyzing the combined effect of a long-term exposure to sodium arsenite, which probably affected the repair mechanism, and of a single dose of TEPA. It is concluded that the present stage of knowledge requires acceptance of the opinion that the genetic risk induced by chronic exposure to a chemical is as serious as that induced by an acute exposure.