Tumour necrosis factor and the lysosomal enzymes of macrophages or macrophage-like cell line

Summary The relationship between tumour necrosis factor (TNF) and macrophages or macrophage-like cell line, especially the lysosomal enzymes was investigated. The serum lysosomal enzymes and LDH activities were increased in proportion to the TNF production even in different strains of mice. Lysosomal enzymes and TNF activity were released into the supernatant of the culture medium of macrophage-enriched peritoneal exudate cells (PEC) or spleen cells derived from Propionibacterium acnes-primed mice after addition of lipopolysaccharide (LPS). After passage through a Sephadex G-10 column, TNF activity could not be detected in the supernatant of these spleen cells after addition of LPS. Also TNF activity could not be detected in the supernatant following destruction of PEC. These results suggest that TNF producibility is strongly related to the degree of activation of macrophages, especially the lysosomal enzymes. The murine macrophage-like cell line, J 774, also released TNF activity and lysosomal enzymes after addition of LPS.     

Protein kinase C may regulate resting anion conductance in vascular smooth muscle cells

The blocker of protein kinase C(PKC) activated large-conductance channel(337.1 pS) in cell attached patch mode in cultured vascular smooth muscle cells. The channel showed time-dependent inactivation whose time course became faster as the amplitude of the command potential was increased. These characteristics of large-conductance channel activated by the application of the PKC blocker were very similar to those of voltage-dependent Cl channels in these cells, indicating the channel activated by the drug is Cl channel. Since voltage-dependent Cl channels were reported to be only activated by forming inside-out patch, these findings suggest Cl permeability of vascular smooth muscle cells is at least partially regulated by protein kinase C.     

The Amino Acid Content in Gamma-irradiated Rice†

High phosphate accumulating bacteria were isolated by autoradiography. One isoate, Arthrobacter globiformis PAB-6 accumulated phosphate intracellularly at 20% of dry cell mass in a simple synthetic medium. This amount was 3~7 times higher than type cultures examined. Almost no phosphate was released into the medium after cessation of growth. Fifty percent of total intracellular phosphate was fractionated as nucleic acids, while 20% each was recovered from cold PCA soluble fractions and polyphosphate fractions. The large content of nucleic acids in this bacterium appeared due to increased RNA content, specifically 4 S RNA fraction.     

Changes in Lysozyme due to Interaction with Vaporized Hexanal

In order to elucidate the mechanism of the alteration of proteins induced by vaporized aldehydes, unmodified and chemically-modified lysozymes were exposed in the solid state to vaporized hexanal at 50°C and 5.8 or 75% relative humidity (RH). On exposure at 75%RH, the unmodified lysozyme exhibited polymerization, browning, loss of solubility, fluorescence production and impairment of lysine, tryptophan and methionine residues. Methionine residues seemed to be oxidized to methionine sulfoxide residues. The polymerization did not proceed at 5.8RH. All the above alterations were almost completely prevented by the removal of oxygen from the reaction cells. Acetylation of lysozyme retarded these alterations fairly well except that the impairment of tryptophan residues was unaffected.

On the basis of all the results it is suggested that at the first step the concerned reaction essentially requires hexanal derivatives such as peroxyhexanoic acid and/or related radicals induced through the reaction with oxygen. The second step seems to consist at least of two routes which are independent of each other and require water. One route is assumed to be an amino-carbonyl reaction involving lysine residues. The other route seems responsible for the attack on tryptophan and methionine residues through oxidation involving the radicals.     

Diversity and host associations of dipteran insects exploiting fungal fruiting bodies in Hokuriku,central Japan

To clarify the diversity and host associations of dipteran insects exploiting fungal fruiting bodies, we collected fruiting bodies at 18 localities in Hokuriku region, central Japan, from 2012 to 2015 and examined them for the emergence of insects. In total, 14,107 dipteran individuals belonging to 20 families emerged from fungi of 8 orders, 25 families, 49 genera and 129 species. Approximately 79% of dipteran individuals belonged to three families, Phoridae, Muscidae and Drosophilidae. The faunal similarity at the family level was relatively high between central (warm‐temperate) and northern (cool‐temperate) areas of Japan. However, the species composition of Drosophilidae was much different between central and northern Japan. The difference in the species composition was discussed in relation to the climatic conditions and fungal flora. None of the species from Drosophilidae, Phoridae, Muscidae, Mycetophilidae, Lonchaeidae and Chloropidae were specialists (they exploited more than one species of fungi), but they showed differences their fungi preference. Adults of some families, especially Drosophilidae, were frequently collected from fruiting bodies, but those of other families were seldom collected, probably reflecting differences in adult feeding ecology.     

Polyglutamylated Tubulin Binding Protein C1orf96/CSAP Is Involved in Microtubule Stabilization in Mitotic Spindles

The centrosome-associated C1orf96/Centriole, Cilia and Spindle-Associated Protein (CSAP) targets polyglutamylated tubulin in mitotic microtubules (MTs). Loss of CSAP causes critical defects in brain development; however, it is unclear how CSAP association with MTs affects mitosis progression. In this study, we explored the molecular mechanisms of the interaction of CSAP with mitotic spindles. Loss of CSAP caused MT instability in mitotic spindles and resulted in mislocalization of Nuclear protein that associates with the Mitotic Apparatus (NuMA), with defective MT dynamics. Thus, CSAP overload in the spindles caused extensive MT stabilization and recruitment of NuMA. Moreover, MT stabilization by CSAP led to high levels of polyglutamylation on MTs. MT depolymerization by cold or nocodazole treatment was inhibited by CSAP binding. Live-cell imaging analysis suggested that CSAP-dependent MT-stabilization led to centrosome-free MT aster formation immediately upon nuclear envelope breakdown without γ-tubulin. We therefore propose that CSAP associates with MTs around centrosomes to stabilize MTs during mitosis, ensuring proper bipolar spindle formation and maintenance.     

Hypergravity Provokes a Temporary Reduction in CD4+CD8+ Thymocyte Number and a Persistent Decrease in Medullary Thymic Epithelial Cell Frequency in Mice

Gravity change affects many immunological systems. We investigated the effects of hypergravity (2G) on murine thymic cells. Exposure of mice to 2G for three days reduced the frequency of CD4⁺CD8⁺ thymocytes (DP) and mature medullary thymic epithelial cells (mTECs), accompanied by an increment of keratin-5 and keratin-8 double-positive (K5⁺K8⁺) TECs that reportedly contain TEC progenitors. Whereas the reduction of DP was recovered by a 14-day exposure to 2G, the reduction of mature mTECs and the increment of K5⁺K8⁺ TEC persisted. Interestingly, a surgical lesion of the inner ear’s vestibular apparatus inhibited these hypergravity effects. Quantitative PCR analysis revealed that the gene expression of Aire and RANK that are critical for mTEC function and development were up-regulated by the 3-day exposure and subsequently down-regulated by the 14-day exposure to 2G. Unexpectedly, this dynamic change in mTEC gene expression was independent of the vestibular apparatus. Overall, data suggest that 2G causes a temporary reduction of DP and a persistent reduction of mature mTECs in a vestibular system-dependent manner, and also dysregulates mTEC gene expression without involving the vestibular system. These data might provide insight on the impact of gravity change on thymic functions during spaceflight and living.