Evaluation of the ability of intact platelets to accumulate acridine orange

A new approach to the evaluation of the uptake of fluorescent probes by intact cells is described. Acridine orange (AO) was used because it can be selectively accumulated by serotonin-containing granules of platelets. Analysis of the fluorescence signal allows the estimation of the relative volume of the granules and the equilibrium coefficients for AO transport across the cytoplasm and granule membranes. The following results were obtained for human and rabbit platelets: the relative volumes of the granules were 14 +/- 1% and 29 +/- 2%, the ratios of intragranular-extracellular probe concentration were 2260 +/- 382 and 30,000 +/- 5550, and the cytoplasm-extracellular medium concentration ratios were 375 +/- 60 and 225 +/- 60, respectively.     

Age distribution of circulatingα-interferon

Summary A sensitive radioimmunoassay showed that circulating -interferon in the plasma of healthy individuals was low in children and reached the highest level in the young adult, then declined gradually with age. Circulating -interferon was 0.201±0.059 ng/ml in males (n=19) and 0.184±0.076 ng/ml in females (n=14) at ages 30–39 years old. It was noted that circulating -interferon was maintained up to a certain level even in elderly individuals.     

De novo mutations in the gene encoding STXBP1 (MUNC18-1) cause early infantile epileptic encephalopathy

Early infantile epileptic encephalopathy with suppression-burst (EIEE), also known as Ohtahara syndrome, is one of the most severe and earliest forms of epilepsy. Using array-based comparative genomic hybridization, we found a de novo 2.0-Mb microdeletion at 9q33.3-q34.11 in a girl with EIEE. Mutation analysis of candidate genes mapped to the deletion revealed that four unrelated individuals with EIEE had heterozygous missense mutations in the gene encoding syntaxin binding protein 1 (STXBP1). STXBP1 (also known as MUNC18-1) is an evolutionally conserved neuronal Sec1/Munc-18 (SM) protein that is essential in synaptic vesicle release in several species. Circular dichroism melting experiments revealed that a mutant form of the protein was significantly thermolabile compared to wild type. Furthermore, binding of the mutant protein to syntaxin was impaired. These findings suggest that haploinsufficiency of STXBP1 causes EIEE.     

Effect of insulin on the synthesis and release of lipid peroxide by cultured hepatocytes isolated from normal and diabetic rats

Lipid peroxide content in hepatocytes isolated from ketotic diabetic rats was higher than normal, and the release of peroxide into the media was also elevated for the initial 18 h. Insulin suppressed both peroxide release and synthesis by cultured hepatocytes isolated from normal and from diabetic rats.     

The scent scale substances of malePieris butterflies (Pieris melete andPieris napi)

Summary 7 monoterpenes, -pinene, -pinene, myrcene, p-cymene, limonene, neral and geranial, were identified together with n-undecane in the scent scales ofP. melete, and also, together with considerable amounts of linalool, in the scent scales ofP. napi. Content ratio of neral to geranial inP. melete ranged from 0.77 to 1.04, and that inP. napi from 1.84 to 2.43. The present chemotaxonomic approach shows the 2 species ofPieris to be distinct.The authors thank Dr T. Yushima (National Institute of Agricultural Science, Japan) for supplying the references 3 and 4.     

Inhibition of IgM antibody-mediated aggregation ofTrypanosoma gambiense in the presence of complement

This paper deals with the immune reaction betweenTrypanosoma gambiense and monoclonal IgM mouse antibody at equivalence with or without rabbit complement. Antibody-mediated trypanosome clumps formed in the absence of complement, and were readily dissociated by complement to become free. In the presence of complement, on the other hand,T. gambiense were not aggregated by the antibody. Free parasites adhered readily to cultured peritoneal macrophages. Complement-mediated dissociation of the clumped trypanosomes in the equivalence area released a large number of previously bound surface antigens. These antigens were capable of binding again to fresh IgM antibody. Experimental results further indicated that the complement system caused a functional alteration, changing the multivalent nature of the IgM antibody in the immune complex into a univalent one. This phenomenon is of great advantage to the infected host in clearing pathogens in vivo, as it allows more antibodies to attach to trypanosomes and subsequently initiate complement activity.     

Synthesis of rhodotorucine A,the inducing factor of mating tube formation ofRhodosporidium toruloides

Summary The inducing factor of mating tube formation ofRhodosporidium turuloides, named rhodotorucine A (H-Tyr-Pro-Glu-Ile-Ser-Trp-Thr-Arg-Asn-Gly-Cys(S-farnesyl)-OH), has been synthesized to confirm the structure proposed for the natural lipopeptide. The synthetic S-farnesyl undecapeptide has identical Rf values on TLC using several different solvents, and also the same biological activity as the natural hormone.Acknowledgments. We wish to express our thanks to Drs E. Ohmura, M. Nishikawa and M. Yoneda of Takeda Chemical Industries and Dr I. Banno of Institute for Fermentation, Osaka, for their encouragement throughout this work.     

Sylvaticin: a new cytotoxic and insecticidal acetogenin from Rollinia sylvatica (Annonaceae)

Sylvaticin (I), a new tetrahydroxy annonaceous acetogenin with nonadjacent tetrahydrofuran rings, has been isolated from the dried fruits of Rollinia sylvatica St. Hil. (Annonaceae). This compound is extremely cytotoxic to human tumor cells and shows promising insect control properties.     

Defining the resistance to oxygen transfer in tissue hypoxia

Studies of O2 supply in freshly isolated adult mammalian cells provide new insight into the factors that limit mitochondrial oxygenation in vivo. Of particular importance, mitochondria are present at high densities and often in apparent clusters, both of which contribute to local O2 gradients under hypoxic conditions. Current evidence indicates that the mitochondrial distribution is a component of the differentiated phenotype of adult mammalian cells and that specific motors and anchoring mechanisms are present to allow redistribution in response to developmental, physiological and pathological challenges. To compare the importance of resistance to O2 transfer under different conditions and at different sites along the supply path in vivo, a simple mathematical expression of relative resistance to O2 supply is introduced. Under various pathophysiological conditions, this resistance increases in specific regions of the pulmonary, circulatory or cellular supply path and results in O2 deficiency in the mitochondria. Regardless of cause, the relative resistance increases dramatically in the vicinity of mitochondrial clusters during hypoxia.