Heteropolypeptides from hydrogen cyanide and water? Solid state15N NMR investigations

Cross-polarization magic-angle spinning¹⁵NMR spectra have been used to determine the composition of hydrogen cyanide polymers both before and after treatment with water. The unambiguous presence of secondary amide groups (as in peptide links) has been established by double-cross-polarization studies on the polymers synthesized from equimolar amounts of H¹³CN and HC¹⁵N. The NMR results are consistent with the hypothesis that the original heteropolypeptides on Earth were synthesized directly from hydrogen cyanide and water without the intervening formation of -amino acids.     

Giardia muris: scanning electron microscopy of in vitro excystation

A recently developed in vitro excystation procedure results in almost total excystation of Giardia muris, an intestinal parasite of mice. The present experiment examines the G. muris cyst morphology by scanning electron microscopy and the efficacy of the excystation procedure. Untreated cysts of G. muris were elliptical and displayed a distinctive surface structure. Excystation began almost immediately after incubation had begun and most trophozoites emerged within 30 min. Excystation appears to involve flagellar action of the encysted trophozoite. A tear of the wall occurred at one pole. This opening was subsequently enlarged, presumably by flagellar action. Trophozoites emerged, posterior end first, and an associated mucoid-like material was extruded. Newly emerged trophozoites were nearly oval in shape. Trophozoites quickly became flattened, elongate, and underwent cytokinesis resulting in two daughter trophozoites. Few organisms not excysted were seen after 30 min incubation.     

Elemental distribution in striated muscle and the effects of hypertonicity: Electron probe analysis of cryo sections

A method of rapid freezing in supercooled Freon 22 (monochlorodifluoromethane) followed by cryoultramicrotomy is described and shown to yield ultrathin sections in which both the cellular ultrastructure and the distribution of diffusible ions across the cell membrane are preserved and intracellular compartmentalization of diffusabler ions can be quantitated. Quantitative electron probe analysis (Shuman, H., A.V. Somlyo, and A.P. Somlyo. 1976. Ultramicros. 1:317-339.) of freeze-dried ultrathin cryto sections was found to provide a valid measure of the composition of cells and cellular organelles and was used to determine the ionic composition of the in situ terminal cisternae of the sarcoplasmic reticulum (SR), the distribution of CI in skeletal muscle, and the effects of hypertonic solutions on the subcellular composition if striated muscle. There was no evidence of sequestered CI in the terminal cisternae of resting muscles, although calcium (66mmol/kg dry wt +/- 4.6 SE) was detected. The values of [C1](i) determined with small (50-100 nm) diameter probes over cytoplasm excluding organelles over nuclei or terminal cisternae were not significantly different. Mitochondria partially excluded C1, with a cytoplasmic/ mitochondrial Ci ratio of 2.4 +/- 0.88 SD. The elemental concentrations (mmol/kg dry wt +/- SD) of muscle fibers measured with 0.5-9-μm diameter electron probes in normal frog striated muscle were: P, 302 +/- 4.3; S, 189 +/- 2.9;C1, 24 +/- 1.1;K, 404 +/- 4.3, and Mg, 39 +/- 2.1. It is concluded that: (a) in normal muscle the "excess CI" measured with previous bulk chemical analyses and flux studies is not compartmentalized in the SR or in other cellular organelles, and (b) the cytoplasmic C1 in low [K](0) solutions exceeds that predicted by a passive electrochemical distribution. Hypertonic 2.2 X NaCl, 2.5 X sucrose, or 2.2 X Na isethionate produced: (a) swollen vacuoles, frequently paired, adjacent to the Z lines and containing significantly higher than cytoplasmic concentrations of Na and Cl or S (isethionate), but no detectable Ca, and (b) granules of Ca, Mg, and P = approximately (6 Ca + 1 Mg)/6P in the longitudinal SR. It is concluded that hypertonicity produces compartmentalized domains of extracellular solutes within the muscle fibers and translocates Ca into the longitudinal tubules.     

An alkali-soluble polysaccharide from the cell walls of Coprinus lagopus

An alkali-soluble polysaccharide was isolated from the purified mycelial walls of Coprinus lagopus. The hydrolysis products, optical rotation, and infrared spectrum indicate a -glucan. Hydrolysis of the glucan after permethylation gave only 2,3,4,6-tetra-, 2,4,6-tri-, and 2,4-di-O-methyl-d-glucose. These methylated sugars and their relative quantities reveal that the glucan is a polysaccharide containing -1,3-linked glucose units with about 14% of the sugars having 1,6-linked branch points. Partial hydrolysis of the product derived from Smith degradation of the glucan released laminaribiose and gentiobiose suggesting that the branches are generally longer than a single glucose unit.Adapted from a portion of a dissertation submitted in partial fullfillment of the requirements for the Ph.D. degree in botany at the University of Connecticut under the direction of Dr. Ralph P. Collins     

In vivo metabolism of a mutant apolipoprotein, apoA-IIowa, associated with hypoalphalipoproteinemia and hereditary systemic amyloidosis.

Apolipoprotein (apo) A-I is the major protein constituent of plasma high density lipoproteins (HDL). A kindred has been identified in which a glycine to arginine mutation at residue 26 in apoA-I is associated with hypoalphalipoproteinemia and hereditary systemic amyloidosis. We isolated the mutant protein, termed apoA-IIowa, from the plasma of an affected subject and studied its in vivo metabolism compared to that of normal apoA-I in two heterozygous apoA-IIowa subjects and two normal controls. Normal and mutant apoA-I were radioiodinated with 131I and 125I, respectively, reassociated with autologous plasma lipoproteins, and simultaneously injected into all subjects. Kinetic analysis of the plasma radioactivity curves demonstrated that the mutant apoA-IIowa was rapidly cleared from plasma (mean fractional catabolic rate [FCR] 0.559 day-1) compared with normal apoA-I (mean FCR 0.244 day-1) in all four subjects. The FCR of normal apoA-I was also substantially faster in the heterozygous apoA-IIowa subjects (mean FCR 0.281 days-1) than in the normal controls (mean FCR 0.203 days-1). Despite the rapid removal from plasma of apoA-IIowa, the cumulative urinary excretion of its associated radioactivity after 2 weeks (44%) of the injected dose) was substantially less than that associated with normal apoA-I (78% of injected dose), indicating extravascular sequestration of radiolabeled apoA-IIowa.(ABSTRACT TRUNCATED AT 250 WORDS)     

Interferon-induced alterations in membrane functions and the growth of Daudi lymphoma and Friend leukemia cells

The Friend murine erythroleukemia cell system and the Daudi Burkitt's lymphoma cell system were used to study the effect of growth-inhibitory concentrations of interferon on membrane functions. Experiments with Friend-cell clones sensitive and resistant to interferon indicated that a number of changes in membrane transport occur rapidly after the addition of interferon to sensitive cells. While no change was observed in the activity of the (Na+/K+) ATPase in Friend cells sensitive or resistant to interferon, a piretanide-inhibitable Na+,K+, 2Cl- co-transport system was specifically inhibited after interferon treatment of sensitive cells. In contrast, treatment of Daudi cells with purified molecularly cloned or standard preparations of human leukocyte interferon gave rise to no early changes in the transport of amino acids, 32Pi, sugars, or 86Rb+. The major change observed in Daudi cells was a marked reduction in the uptake and incorporation of thymidine, which begins to decrease after 8-10 h of exposure to interferon.     

Different repetition frequencies of a 120 base-pair DNA-element and its arrangement in Chironomus thummi thummi and Chironomus thummi piger

The repetition frequency of a highly repetitive DNA sequence has been measured in the genomes of Ch. thummi thummi and Ch. th. piger. This sequence is known to be involved in the evolutionary duplication of defined chromosomal segments leading to a significant increase in the genome size of Ch. th. thummi. Reassociation of this highly repetitive DNA sequence which has a repeat length of 120 base-pairs, with total Ch. th. thummi and Ch. th. piger DNA has shown that the repetition frequency in the Ch. th. thummi DNA is 5.5 fold higher than in Ch. th. piger. In both genomes a 120 base-pair sequence is present as tandemly repeated sequence as shown by Southern analysis.     

Preparation of membrane vesicles from isolated myelin: studies on functional and structural properties.

Myelin membranes purified from bovine brain are shown to form membrane vesicles when incubated in hypotonic buffer. Following restoration of isotonicity a resealing of the membrane occurs as judged by a significant decrease in 22Na+ permeability. Electron spin resonance measurements using stearic acid spin label I indicate a small decrease in membrane fluidity with increasing ionic strength between 50 and 80 mM NaCl. Iodination of myelin membrane vesicles by lactoperoxidase shows a four-fold increase in the amount of iodine incorporation into the myeline basic protein from 0--150 mM NaCl, while the iodination of the proteolipid protein remains essentially unaffected by the change in ionic strength. This dependence of the iodination of the myelin basic protein on the ionic strength can be explained by the electrostatic interactions of this protein with membrane lipids. In view of striking analogies with studies on model membranes correlating protein binding with membrane permeability changes, we suggest a similar structure-function relationship for the myelin basic protein.     

Seasonal fluctuations of food intake and bodyweight in the dormouse Glis glis (author's transl)]

Cycles of body weight lasting a few weeks occurred in dormice fed ad libitum, throughout the year with the same food. The pattern of the fluctuations suggests an oscillation between an upper and a lower limit. When the lower limit is reached, compensatory mechanisms acting on the food intake are induced which cause an increase in body weight to the upper one; when this upper limit is reached, other mechanisms cause the body weight to decrease to the lower one. Thus, body weight and variations in food intake may well be controlled by the difference between the body weight and the limit it will reach. In a warm environment (23-25 degrees C) these fluctuations are smaller than in the cold (5-6 degrees C). They are also smaller during summer than during winter in animals placed in natural climatic conditions. In summer, the body weight stays near the upper limit; in this season, as in the warm environment, the lower limit approaches the upper one. Thus, the main circannual variation of body weight regulation in dormice may be the elevation during summer of the lower body weight limit.