Selenocysteine lyase activity in a cysteine-requiring mutant ofEscherichia coli K-12

Selenocysteine lyase activity was detected in crude extracts from a cysteine-requiring mutant ofEscherichia coli K-12. The level of activity was the same whether cells had been grown aerobically or anaerobically, with or without selenocysteine. Selenocysteine lyase catalyzes the conversion of selenocysteine to alanine and elemental Se, a reaction that is followed by a nonenzymatic reduction of the Se to hydrogen selenide. Both of these end products were identified in this study. With cysteine as the substrate, alanine and H₂S were formed, but only at levels 50% less than the products formed from selenocysteine. Selenocysteine lyase has been identified in a number of mammals and bacteria; its presence in a cysK mutant ofE. coli K-12 suggests a common route whereby hydrogen selenide, derived from selenocysteine, can then be assimilated into selenoproteins.     

High Bulk Diet for Diverticular Disease of the Colon

In the past century diverticular disease of the colon has changed from being almost unknown to becoming the most common disease of the colon. Studies in Britain indicated that the pathological basis of the disease is a thickening of the colonic musculature, with diverticulosis and diverticulitis developing because of increased intracolonic pressures generated by the thickened colon wall. This pressure can be sharply reduced by increased colonic bulk. Geographical and anthropological data reveal that diverticular disease results from Western civilization''s food habits, specifically the reduced fiber content in food. There is evidence that increasing the dietary intake of fiber by the addition of bran can prevent formation of diverticula and relieve the symptoms of established disease. Large scale studies are recommended both as treatment and to further test the validity of this concept.     

Branched-chain glycosyl α-amino acids : Part II. Synthesis of 2-D- and 2-L-(3-deoxy-1,2:5,6-di-O-isopropylidene-α-D-glucofuranos-3-yl)glycine. Analogs of the sugar moiety of the polyoxins

Stereospecific hydroxylation of 3-deoxy-1,2:5,6-di-O-isopropylidene-3-C-trans-and 3-C-cis-(methoxycarbonylmethylene)-α-D-ribo-hexofuranose (2 and 3, respectively), with potassium permanganate in pyridine afforded 3-C-[S- and R-hydroxy-(methoxycarbonyl)methyl]-1,2:5,6-di-O-isopropylidene-α-D-glucofuranose, (6 and 7, respectively), in a combined yield, after chromatography, of 43%. Selective formation of monomethanesulfonates (9a and 10a) and p-toluenesulfonates (9b and 10b), followed by treatment with sodium azide and reduction of the azide, afforded the methyl 2-D-(and 2-L-)(3-deoxy-1,2:5,6-di-O-isopropylidene-α-D-glucofuranos-3-yl)-glycinates (12a and 13a, respectively). Basic hydrolysis of the latter compounds yielded 2-D- and 2-L-(3-deoxy-1,2:5,6-di-O-isopropylidene-α-D-glucofuranos-3-yl)glycine (12b and 13b, respectively). The structures of the glycosyl amino acids were correlated with that of L-alanine by circular dichroism.     

DISTRIBUTION OF PEROXISOMES (MICROBODIES) IN THE NEPHRON OF THE RAT : A Cytochemical Study

The distribution of peroxisomes (microbodies) in the rat nephron was studied cytochemically, using glutaraldehyde- or formaldehyde-fixed tissue, by means of α-hydroxy acid oxidase activity in light microscopy or oxidation of 3,3'-diaminobenzidine (DAB) at pH 9 in both light and electron microscopy.The two cytochemical methods show peroxisomes to be nearly sperical particles found only in cells of the proximal convoluted tubule. Lysosomes were identified in the same or parallel sections, with β-glycerophosphate or 5'-cytidylic acid as substrate. They are found in all cells of the nephron. These cytochemical methods visualize the two organelles for light microscopy; they also permit unequivocal differentiation of all kidney peroxisomes from lysosomes in electron micrographs. Peroxisomes are larger and more reactive in the cells of the pars descendens (P₃ segment) of the proximal convolution, located in the outer medulla and medullary rays, than in the cells of the pars convoluta (P₁ and P₂ segments), situated in the cortex. In contrast, lysosomes are much smaller in the P₃ segment and larger and more reactive in the P₁ and P₂ segments. In all cells of the proximal convolution, peroxisomes tend to be concentrated nearer the base of the cells than do lysosomes. Mitochondria in P₃ cells also show low levels of DAB oxidation at pH 6, in contrast to those in P₁ and P₂ cells. The possibility is discussed that P₃ cells possess an extramitochondrial means of oxidation in which peroxisome oxidases play an important role.     

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).     

LYSOSOMES IN THE RAT SCIATIC NERVE FOLLOWING CRUSH

Peripheral nerves undergoing degeneration are favorable material for studying the types, origins, and functions of lysosomes. The following lysosomes are described: (a) Autophagic vacuoles in altered Schwann cells. Within these vacuoles the myelin and much of the axoplasm which it encloses in the normal nerve are degraded (Wallerian degeneration). The delimiting membranes of the vacuoles apparently form from myelin lamellae. Considered as possible sources of their acid phosphatase are Golgi vesicles (primary lysosomes), lysosomes of the dense body type, and the endoplasmic reticulum which lies close to the vacuoles. (b) Membranous bodies that accumulate focally in myelinated fibers in a zone extending 2 to 3 mm distal to the crush. These appear to arise from the endoplasmic reticulum in which demonstrable acid phosphatase activity increases markedly within 2 hours after the nerve is crushed. (c) Autophagic vacuoles in the axoplasm of fibers proximal to the crush. The breakdown of organelles within these vacuoles may have significance for the reorganization of the axoplasm preparatory to regeneration. (d) Phagocytic vacuoles of altered Schwann cells. As myelin degeneration begins, some axoplasm is exposed. This is apparently engulfed by the filopodia of the Schwann cells, and degraded within the phagocytic vacuoles thus formed. (e) Multivesicular bodies in the axoplasm of myelinated fibers. These are generally seen near the nodes of Ranvier.     

An Association between Mitochondria and the Endoplasmic Reticulum in Cells of the Pseudobranch Gland of a Teleost

An elaborate and apparently unique specialization of the endoplasmic reticulum having the form of tubules and a precise orientation with respect to the mitochondria has been described for the specific cell of the pseudobranch gland. The tubules also are concentrated near the vascular border of the cell where they show continuity with the plasma membrane and open directly against the basement membrane. On the other side of the basement membrane, the endothelial cells of the sinusoid show openings or discontinuities characteristically associated with secretory cells. The pseudobranch gland is presumed to have carbonic anhydrase as one of its primary products, if not its only one, and the elaborate ultrastructure is thought to be associated with the special problems of secreting this enzyme.     

Barbiturate Mortality as an Index of Barbiturate Use,Canada, 1950-1963

Changes in Canadian rates of mortality from barbiturates are examined, and their relation to barbiturate use in the general population is discussed. While the number of deaths attributed to barbiturates quadrupled, from 63 in 1950 to 232 in 1963, there has been a concomitant decrease in the number of deaths from inhalation of utility gas.Combined rates for deaths from utility gas and barbiturates declined steadily for most age groups between 1950-52, 1955-57, and 1959-63. It is possible that the increased mortality from barbiturates represents a change in fashion in regard to method of suicide. Changed mortality from barbiturates is not a valid measure of the extent to which consumption of barbiturates has increased in the Canadian population.