Isolation and characterization of a type II restriction endonuclease from Streptococcus thermophilus

The alpha-toxin (phospholipase C) of Clostridium perfringens has been reported to contain catalytically essential zinc ions. We report here that histidine residues are essential for the co-ordination of these ion(s). Incubation of alpha toxin with diethylpyrocarbonate, a histidine modifying reagent, did not result in the loss of phospholipase C activity unless the protein was first incubated with EDTA, suggesting that zinc ions normally protect the susceptible histidine residues. When the amino acid sequences of three phospholipase C's were aligned, essential zinc binding histidine residues in the non-toxic B. cereus phospholipase C were found in similar positions in the toxic C. perfringens enzyme and the weakly toxic C. bifermentans phospholipase C.     

Partial purification and characterization of cysteine proteinases from various developmental stages of Paragonimus westermani

1. During development of Paragonimus westermani, larvae develop during migration within the host, and adult worms feed on pulmonary tissues, causing significant pathology in the mammalian host. In this report acidic extracts of various developmental stages (metacercariae and worms at one, two and three months of development) were examined for cysteine proteinase activity. 2. A soluble thiol-dependent proteinase activity with a native molecular weight of approximately 20,000 was isolated and partially purified. 3. The enzymes purified from the various developmental stages of the parasite had maximal activity at acidic pH and showed inhibitor susceptibilities similar to the vertebrate acidic cysteine proteinases. 4. Enzymatic activity was stable at pH 5.0 for at least two days when stored at 4 degrees C. 5. It is suggested that these enzymes may be involved in the nutrition of these parasites and/or during penetration and lysis of the tissues.     

Simulation study on effects of channel noise on differential conduction at an axon branch.

Effects of membrane channel noise (random opening and closing of ion channels) are studied on spike conduction at a branching point on an axon. Computer simulation is done on the basis of a stochastic version of the Hodgkin-Huxley cable model, into which the channel noise is incorporated. It is shown that the channel noise makes conduction of spikes into daughter branches random; spikes randomly succeed or fail in conduction into daughter branches. The conduction is then randomly differential even though the forms and properties of daughter branches are the same. The randomness is considerable when the radius of an axon is small (approximately 1 microns).     

Significance of exercise and bed rest in pregnancy--study on the lying postures of gravidas during sleep (2)

For the purpose of determining the most reasonable lying posture for pregnant women, we investigated the lying positions of both 247 non-pregnant women and 302 pregnant women during sleep. As for the rate of each position during the entire period of observation, 33.2% of the non-pregnant women were in the supine position, 41.2% in the lateral position, 18.4% in Sims' position and 7.1% in the prone position. In the pregnant group, the rate of supine position, simple lateral position and Sims' position was 34.2%, 52.2% and 12.7%, respectively, but the rate of the prone position was limited to 0.8%. All of the gravidas assuming the prone position were at less than 16 weeks of gestation. Non-pregnant women could sleep in a variety of positions, but pregnant women could assume the prone position during sleep only in the stage when the abdomen is not yet prominent or distended. Pregnant women were restricted significantly, either consciously or unconsciously, with progress in gestation.     

Thermotropic phase behavior and stability of monosialoganglioside micelles in aqueous solution.

The thermotropic phase behavior of monosialoganglioside in a dilute aqueous dispersion at pH 6.8 was measured by using synchrotron radiation small-angle x-ray scattering and was analyzed by a shell-modeling method. Previous calorimetric studies on ganglioside systems have shown quite different thermotropic behaviors from other biological lipid systems, however, the details have still been ambiguous. Because of high statistical data and a shell-modeling analysis, we could elucidate the internal structural change of monosialoganglioside micelle induced by the elevation of temperature from 6 to 60 degrees C, that is, the shrinkage of the hydrophilic region and the slight expansion of the hydrophobic region occurring simultaneously, accompanying the elongation of the axial ratios of the ellipsoidal micelles. The model structures obtained explain the changes in the experimental scattering curves, the distance distribution functions, and the gyration radii. In addition we have also found an evident thermal hysteresis in the scattering curves and in the structural parameters. The present result suggests that the thickness of the hydrophilic region, namely, the conformation of oligosaccharide chains, is sensitive to a change of temperature.     

Ubiquitin metabolism in HeLa cells starved of amino acids.

Radio-iodinated ubiquitin (Ub) was introduced into HeLa cells by red blood cell-mediated microinjection. The half-life and solubility of Ub, as well as the molecular weight distributions of Ub conjugates, were then measured in HeLa cells grown in complete medium or in medium lacking amino acids and fetal calf serum. Ub metabolism was similar in the two sets of cells. Thus, the dramatic changes in Ub metabolism induced by thermal stress are not observed upon amino acid deprivation.     

Methylation patterns at the hypervariable X-chromosome locus DXS255 (M27 beta): correlation with X-inactivation status.

Methylation patterns surrounding a hypervariable X-chromosome locus, DXS255, have been analyzed with the restriction enzyme MspI and its methylation-sensitive isoschizomer HpaII. HpaII sites flanking the hypervariable region were found to be methylated on 41 active X chromosomes and unmethylated on 11 inactive X chromosomes present in a range of male, female, and hybrid cells and tissues. This differential methylation pattern coupled with the previously described high level (greater than 90%) of heterozygosity at the DXS255 locus can therefore be applied to determine the inactivation status of X chromosomes in females heterozygous for X-linked disease and in tumor clonality studies.