Effect of streptococcal pyrogenic exotoxin on rabbit macrophage functions in vitro: mediation by splenic lymphocytes

Streptococcal pyrogenic exotoxin (SPE) showed no direct effect on rabbit macrophage functions in vitro. However, when splenic lymphocytes were added to macrophage cultures, SPE caused marked augmentation of glucose consumption and superoxide anion production, and concomitant inhibition of phagocytosis without loss of cell viability. The SPE effects were demonstrated to be mediated by a soluble factor(s) released from the splenic lymphocytes in response to SPE stimulus.     

Generation of inositol phosphates, cytosolic Ca and secretion of noradrenaline in PC12 cells treated with glutamate

Glutamate transiently stimulated rat pheochromocytoma PC12 cells and caused an inositol trisphosphate formation and an increase in levels of Ca⁺ in the cytosol. The rank order of potency of glutamate> N-methyl-D-aspartate (NMDA) > KAINATE = quisqualate is characteristic of an interaction with NMDA receptors. The effect of glutamate on inositol trisphosphate formation disappeared in a low Mg²⁺ buffer and was not blocked by DL-2-amino-5-phosphonovalerate, an antagonist for NMDA receptors coupled to ion channels. Although glutamate failed to stimulate noradrenaline secretion, glutamate enhanced the effect of bradykinin, but not of Ca ionophore A23187, or KC1. These results suggest the existence of metabotropic glutamate receptors, different from previously reported receptors, in PC12 cells.     

Capability of intensity discrimination in multiple receptor model of insect taste

A simple model in which one CNS taste neuron integrates inputsfrom multiple peripheral receptors is introduced in order toinvestigate the mechanism of acute intensity taste discriminationin flies. Information theory is applied to evaluate the acuteability of discrimination provided by the model. The presentanalysis is carried out under two statistical conditions concerningthe uncertainty of receptor response. Based on experimentaldata obtained by Smith et al. (1983), we estimate the mutualinformation entropy of the model. The numerical results obtainedhere indicate that the uncertainty observed in a single receptorresponse is dramatically reduced by the central integration.Furthermore, each of the eleven stimulus intensity levels experimentallyapplied by Smith et al., can be discriminated completely byintegrating the responses of the realistic number of receptors(33–212). Such a great improvement of the differentialsensitivity in the model resolves the discrepancy between thelow differential sensitivity of single sugar receptors (Smithet al., 1983) and the high sensitivity obtained in the feedingbehavior (Dethier and Rhoades, 1954; Dethier and Bowdan, 1984)of the blowfly.     

Correlation between the leaf turnover rate and anti-herbivore defence strategy (balance between ant and non-ant defences) amongst ten species of <Emphasis Type="Italic">Macaranga</Emphasis> (Euphorbiaceae)

We measured variation in the intensities of ant and non-ant anti-herbivore defences amongst ten Macaranga species in Sarawak, Malaysia. Intensities of non-ant defences were estimated by measuring effects of fresh leaves (provided as food) of these Macaranga species on survival of common cutworm larvae [Spodoptera litura (Fabricius), Lepidoptera: Noctuidae]. Intensities of ant defences were estimated by measuring ant aggressiveness in the presence of artificial damage inflicted on plants. As part of our examination of non-ant defences, we measured leaf toughness (punch strength, by penetrometry), and the contents of total phenols and condensed tannin. We demonstrated interspecific variation in intensities of both ant and non-ant defences amongst ten Macaranga species and showed that the rank order of ant defence intensity was negatively correlated with the intensity of non-ant defence. We also found that the balance between ant and non-ant defence intensity was correlated with the rates of leaf turnover and shoot growth. Species investing more in ant defence tended to have higher leaf turnover rates. Macaranga species that occur preferentially in shadier microhabitats had lower leaf turnover rates, suggesting that non-ant defences are more cost-effective in more shade-tolerant species. Our results also suggest that the total intensity of non-ant defences is positively correlated with both leaf toughness and total phenol content.     

Mitochondrial phospholipid hydroperoxide glutathione peroxidase suppresses apoptosis mediated by a mitochondrial death pathway.

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a key enzyme in the protection of biomembranes exposed to oxidative stress. We investigated the role of mitochondrial PHGPx in apoptosis using RBL2H3 cells that overexpressed mitochondrial PHGPx (M15 cells), cells that overexpressed non-mitochondrial PHGPx (L9 cells), and control cells (S1 cells). The morphological changes and fragmentation of DNA associated with apoptosis occurred within 15 h in S1 and L9 cells upon exposure of cells to 2-deoxyglucose (2DG). The release of cytochrome c from mitochondria was observed in S1 cells after 4 h and was followed by the activation of caspase-3 within 6 h. Overexpression of mitochondrial PHGPx prevented the release of cytochrome c, the activation of caspase-3, and apoptosis, but non-mitochondrial PHGPx lacked the ability to prevent the induction of apoptosis by 2DG. An ability to protect cells from 2DG-induced apoptosis was abolished when the PHGPx activity of M15 cells was inhibited by diethylmalate, indicating that the resistance of M15 cells to apoptosis was indeed due to the overexpression of PHGPx in the mitochondria. The expression of members of the Bcl-2 family of proteins, such as Bcl-2, Bcl-xL, Bax, and Bad, was unchanged by the overexpression of PHGPx in cells. The levels of hydroperoxides, including hydrogen and lipid peroxide, in mitochondria isolated from S1 and L9 cells were significantly increased after the exposure to 2DG for 2 h, while the level of hydroperoxide in mitochondria isolated from M15 cells was lower than that in S1 and L9 cells. M15 cells were also resistant to apoptosis induced by etoposide, staurosporine, UV irradiation, cycloheximide, and actinomycin D, but not to apoptosis induced by Fas-specific antibodies, which induces apoptosis via a pathway distinct from the pathway initiated by 2DG. Our results suggest that hydroperoxide, produced in mitochondria, is a major factor in apoptosis and that mitochondrial PHGPx might play a critical role as an anti-apoptotic agent in mitochondrial death pathways.     

Analysis of Nuclear Accumulation of Influenza Nucleoprotein Antigen in the Presence of p-Fluorophenylalanine

When p-fluorophenylalanine (FPA) was added to influenza virus RI/5+-infected cells 4 hr after infection, virus-specific proteins were synthesized but infectious progeny virus was not produced. In these cells, synthesis of viral RNA was strongly inhibited and nucleoprotein (NP) antigen was found predominantly in the nucleus in contrast to untreated cells in which NP antigen was distributed throughout the whole cell. The intracellular location and migration of NP were examined by isotope labeling followed by fractionation of infected cells. In untreated cells, a large portion of the NP was present in the cytoplasm and most of it was detected in the form of ribonucleoprotein (RNP). In contrast, in FPA-treated cells little viral RNP was detectable and NP was present predominantly in the nucleus in a nonassembled, soluble form. When FPA was removed from the culture, synthesis of viral RNA was soon restored and a large amount of viral RNP appeared in the cytoplasm; this was followed by the production of infectious virus. The results of the experiments suggest that the NP synthesized in the presence of FPA is not assembled into viral RNP because of the lack of available RNA, and such NP migrates readily into the nucleus and accumulates there.     

Mitogenic Activity of Cytoplasmic Membranes Isolated from L-Forms of Staphylococcus aureus

Cytoplasmic membranes of L-forms of Staphylococcus aureus exerted a strong mitogenic effect on splenocytes of athymic nude mice as well as normal mice, while a cytoplasmic fraction of the same bacteria did not show definite mitogenicity. The mitogenic principle(s) of the membrane fraction was resistant to treatment with trypsin and was heat stable (at 100 C for 10 min). The active principle(s) in the insoluble residue of the membrane fraction digested with trypsin was not extracted with cold acetone, but could be solubilized by extraction with a cold chloroform-methanol mixture (2:1, v/v). The mitogenic principle(s) in the extract was fractionated by silicic acid column chromatography. Among five fractions separated by chromatography, fractions eluted with chloroform-methanol mixtures (1:1 and 1:20, v/v) were found to be strongly mitogenic. The cytoplasmic membranes of the L-forms also exerted a definite mitogenic effect on guinea pig splenocytes, but not on the thymocytes.     

Induction of erythroid differentiation by cytoplast fusion in mouse erythroleukemia (Friend) cells

An intracellular activity, which is induced by dimethyl sulfoxide (DMSO) or hexamethylenebisacetamide (HMBA) and leads to erythroid differentiation in mouse Friend cells, was characterized by cell fusion between genetically marked intact cells and cytoplasts. For this, a procedure for rapid selection of cybrids was devised by sensitizing non-fused cells with oligomycin. We were able to demonstrate that cytoplasts derived from DMSO- (or HMBA)-treated cells trigger erythroid differentiation upon fusion with UV-irradiated cells. The activity in the cytoplasts remained only transiently and its induction was inhibited by biologically active phorbol esters or cycloheximide. The activity, however, was not induced in cytoplasts by directly treating them with DMSO (or HMBA). These results indicate that (1) the intracellular erythroid-inducing activity is located in cytoplasts, (2) it acts in trans and induces erythroid differentiation as a dominant factor and (3) its production requires de novo nuclear protein synthesis. The mechanisms of the induction of the intracellular activity and of how it triggers erythroid differentiation are discussed.     

Body mass and prostatic cancer: a prospective study.

Previous studies have suggested that increased body mass is associated with an increased risk of prostatic cancer, but these studies have been limited by the fact that they were based on a few simple measurements such as height and weight. Similar results were found in a prospective study of the incidence of prostatic cancer in a cohort of Japanese men born in 1900-19 and living in Hawaii. Further evaluation of the extensive anthropomorphic measurements made in this cohort suggested that the association between measures of body mass and prostatic cancer might be accounted for more by lean tissue than by fat tissue. There was a significant positive association of the risk of prostatic cancer with area of muscle in the arm but not with area of fat in the arm. Further research is needed on the biological mechanisms of carcinogenesis that may be related to both lean and fat tissue and the development of prostatic cancer.