Enhanced liver metabolism of mutagens and carcinogens in fish living in polluted seawater.

Specimens of the seawater fish annular seabream (Diplodus annularis) were caught from a polluted harbor area and from a clean reference area. Seawater concentrates and fish-muscle extracts were not mutagenic in the Salmonella reversion test. Liver preparations of fish from the 2 sources were comparatively assayed for microsomal mixed-function oxidases and cytosolic biochemical parameters, as well as for the ability of S12 fractions to activate promutagens or to detoxify direct-acting mutagens. A shift of the cytochrome P-450 peak from 450.3 to 448.5 was accompanied by a 4.5-fold increase in arylhydrocarbon hydroxylase activity in fish living in the polluted environment. At the same time, glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase were doubled in the cytosol of the same animals, while reduced glutathione (GSH) peroxidase and GSH S-transferase were slightly yet significantly depressed. No significant difference was recorded for other biochemical parameters, including GSH, oxidized glutathione (GSSG) reductase, NADH- and NADPH-dependent diaphorases, and DT diaphorase. In parallel, fish exposed to polluted seawater exhibited a significant and marked enhancement of the metabolic activation of the pyrolysis product Trp-P-2 and of benzo[a]pyrene-trans-7,8-diol, and at the same time were less efficient in detoxifying the antitumor compound ICR 191. Liver S12 fractions from both sources efficiently decreased the direct mutagenicity of sodium dichromate, and failed to activate benzo[a]pyrene and aflatoxin B1 to mutagenic metabolites. These results provide evidence that both biochemical parameters and the overall capacity of fish liver to activate or detoxify certain mutagens can be assumed to be sensitive indicators of exposure to mixed organic pollutants in the marine environment.     

Influence of dietary deficiency of nicotinamide on lead toxicity in young rats

The influence of dietary nicotinamide deficiency on lead intoxication in young developing rats was investigated. The Pb induced an increase in brain dopamine and noradrenaline, inhibition in blood δ-aminolevulinic acid dehydratase activity, an elevation in urinary excretion of δ-aminolevulinic acid and blood and tissue uptake of Pb were significantly more marked in animals maintained on a nicotinamide-deficient diet than those fed a nicotinamide-sufficient diet. The nicotinamide deficiency may enhance the susceptibility to Pb intoxication possible by enhancing the absorption of Pb and altering nicotinic acid metabolism.     

A DEVELOPMENTAL STUDY OF THE LEAVES OF NICOTIANA GLUTINOSA AS RELATED TO THEIR SMOG–SENSITIVITY

Glater , Ruth Bobrov , Richard A. Solberg , and Flora M. Scott . (U. California, Los Angeles, and Los Angeles County Air Pollution Control District.) A developmental study of the leaves of Nicotiana glutinosa as related to their smog-sensitivity. Amer. Jour. Bot. 49(9): 954–970. Illus. 1962.—Plants growing in the fields of Los Angeles County as well as those experimentally fumigated in the laboratory show gross markings in response to smog which vary from species to species, from a glistening appearance of the leaf undersurface due to a temporary accumulation of water in the affected cells through complete necrosis. In dicotyledonous leaves, “silvering,” “bronzing,” brown-black mottling or an increase in anthocyanin may be seen. In monocotyledons, transverse banding, tan in color, or longitudinal streaking of leaves are the usual responses. This damage appears in a characteristic pattern on the leaves, different from that produced by other phytotoxicants. Nicotiana glutinosa plants were grown in the air-filtered greenhouses at UCLA. The normal anatomical development of the foliage was studied and correlated with its susceptibility to smog injury. On a given plant, leaves of different ages show damage in different positions. Very young leaves at the apex of the plant and old leaves at the base of the plant are not sensitive. Expanding leaves between young and old in age are sensitive; in this group a distinct pattern of damage is discernible. Damage markings in the youngest leaves appear only at the tip; in leaves somewhat older, close to midblade; in fully mature leaves, only at the base. This localization of damage is shown to be correlated with the gradient of cellular differentiation from tip toward base as the leaf matures. Those cells which have just attained maximum size (young mature) are sensitive; damage, therefore, is a function of cellular development and maturity. The following anatomical details were analyzed: (1) differentiation and distribution of stomata and their opening and closing on both upper and lower epidermal surfaces and (2) development of intercellular air spaces in palisade and spongy parenchyma tissue. These studies indicate that damage occurs in the region of the leaf where stomata have just become functional and ambient polluted air can make direct contact with interior leaf tissues by virtue of large substomatal chambers and intercellular air spaces.     

Using stable MutS dimers and tetramers to quantitatively analyze DNA mismatch recognition and sliding clamp formation

The process of DNA mismatch repair is initiated when MutS recognizes mismatched DNA bases and starts the repair cascade. The Escherichia coli MutS protein exists in an equilibrium between dimers and tetramers, which has compromised biophysical analysis. To uncouple these states, we have generated stable dimers and tetramers, respectively. These proteins allowed kinetic analysis of DNA recognition and structural analysis of the full-length protein by X-ray crystallography and small angle X-ray scattering. Our structural data reveal that the tetramerization domains are flexible with respect to the body of the protein, resulting in mostly extended structures. Tetrameric MutS has a slow dissociation from DNA, which can be due to occasional bending over and binding DNA in its two binding sites. In contrast, the dimer dissociation is faster, primarily dependent on a combination of the type of mismatch and the flanking sequence. In the presence of ATP, we could distinguish two kinetic groups: DNA sequences where MutS forms sliding clamps and those where sliding clamps are not formed efficiently. Interestingly, this inability to undergo a conformational change rather than mismatch affinity is correlated with mismatch repair.