Measurement of Methionine-Enkephalin [Arg6,Phe7] in Rat Brain by Specific Radioimmunoassay Directed at Methionine Sulphoxide Enkephalin[Arg6,Phe7]

Abstract: A specific and sensitive radioimmunoassay procedure for Metenkephalin[Arg⁶,Phe⁷] which allows its measurement in regions of the rat brain is described. The antiserum was raised against the methionine sulphoxide derivative of the peptide, and all samples and standards were oxidized with hydrogen peroxide prior to use in the assay with chloramine T-oxidized ¹²⁵I-labelled Met(O)-enkephalin[Arg⁶,Phe⁷]. The only significant cross-reactivity was 30% with the reduced heptapeptide Met-enkephalin[Arg⁶,Phe⁷]. The assay showed less than 0.15% cross-reactivity with fragments of the heptapeptide and with leucine-enkephalin-containing peptides. Acid acetone extraction of rat striatum followed by Sephadex G-50 chromatography and reverse-phase high pressure liquid chromatography showed that essentially all immunoreactivity co-chromatographed with Met-enkephalin[Arg⁶,Phe⁷]. This confirmed the specificity of the assay and showed that the striatum does not contain a high concentration of larger molecular weight forms with the heptapeptide at the COOH terminus. Distribution of the heptapeptide followed that of methionine enkephalin, with highest concentrations in the globus pallidus, intermediate levels in caudate-putamen and hypothalamus, and low levels in cortex and cerebellum.     

Alterations in plasma-membrane functions after poliovirus infection

After exposure of HeLa cells to poliovirus there is a rapid decline (within minutes) in fluorescence polarization of DPH (1,6-diphenyl-1,3,5-hexatriene). Within one hour after infection the (Na+/K+)ATPase activity of an isolated plasma-membrane-rich fraction is enhanced, the cell volume decreases, and the intracellular concentration of a potent low-molecular-weight inhibitor of host protein synthesis increases.     

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.     

Transfer RNA and protein synthesis

The three-dimensional structure of yeast phenylalanine transfert RNA has been determined in orthorhombic crystals. The current status of this work is reviewed together with the relationship of the transfer RNA structure in the crystal to its biologically active form. In addition some speculations are put forward regarding the mode of interaction of tRNA molecules in the ribosome and the manner in which tRNA interacts with aminoacyl synthetase.