Delta-tubulin is a component of intercellular bridges and both the early and mature perinuclear rings during spermatogenesis

Mammalian spermatogenesis involves drastic morphological changes leading to the development of the mature sperm. Sperm development includes formation of the acrosome and flagellum, translocation of nucleus-acrosome to the cell surface, and condensation and elongation of the nucleus. In addition, spermatogenic cell progenies differentiate as cohorts of units interconnected by intercellular bridges. Little is known about the structural components involved in the establishment of conjoined spermatogenic cells and the mechanism of nuclear shaping of the male gamete. We identified two isoforms of delta-tubulin and found that the long isoform is predominantly expressed in testis, while the short isoform is expressed in all tissues examined. We also found that delta-tubulin forms intercellular bridges conjoining sister spermatogenic cells. In addition, delta-tubulin is a component of the perinuclear ring of the manchette, which acts on translocation and elongation of the nucleus. Furthermore, small rings clearly distinct from the intercellular bridges, which might mature to perinuclear ring of the manchette in later stages of spermatogenesis, were detected on the cell surface of round spermatids. These results suggest that delta-tubulin is a component of two types of ring, the intercellular bridges and the perinuclear rings, which may be involved in morphological changes of spermatid to mature sperm.     

Amyloid beta peptide (Abeta42) activates PLC-delta1 promoter through the NF-kappaB binding site

The abnormal deposition of amyloid beta peptide (Abeta) is a hallmark of Alzheimer's disease (AD). Phospholipase C-delta1 (PLC-delta1) is also known to abnormally accumulate in the brains of AD patients, but no report has addressed the relationship between these two events. This study investigated the effect of Abeta42 on the PLC-delta1 expression in human neuroblastoma cell lines. The PLC-delta1 mRNA level was increased by treatment with Abeta42 in a RT-PCR analysis. In the reporter assay, Abeta42 was found to activate the PLC-delta1 promoter activity in a dose-dependent manner. A novel NF-kappaB binding site in the PLC-delta1 promoter appeared to be responsible for the Abeta42 activity. First, the dominant negative forms of the NF-kappaB activating molecules, dominant negative TGF-beta activated kinase 1 (dnTAK1) and dnNIK (dominant negative NF-kappaB-inducing kinase), abolished the Abeta42 activity in the reporter assay. Second, the Abeta42 augmented a factor binding on the NF-kappaB site in the electrophoretic mobility shift assay (EMSA), which was abolished by a molar excess of the unlabeled consensus NF-kappaB oligonucleotide. These results suggest that the PLC-delta1 promoter is under the control of NF-kappaB, which mediates the expression of PLC-delta due to the Abeta42 treatment.     

Aluminum-induced oxidative stress in rat brain: response to combined administration of citric acid and HEDTA

Aluminum, a known neurotoxic substance, has been suggested as a contributing factor in the pathogenesis of Alzheimer's disease. Therapeutic efficacy of combined administration of citric acid (CA) and N-(2-hydroxyethyl) ethylenediaminetriacetic acid (HEDTA) was evaluated in decreasing blood and brain aluminum concentration and parameters indicative of hematological disorders and brain oxidative stress. Adult male wistar rats were exposed to drinking water containing 0.2% aluminum nitrate for 8 months and treated once daily for 5 consecutive days with CA (50 mg/kg, orally) or HEDTA (50 mg/kg, intraperitoneally) either individually or in combination. Aluminum exposure significantly inhibited blood delta-aminolevulinic acid dehydratase while increased zinc protoporphyrin confirming changed heme biosynthesis. Significant decrease in the level of glutathione S-transferase in various brain regions and an increase in whole brain thiobarbituric acid reactive substance, and oxidized glutathione (GSSG) levels were also observed. Glutathione peroxidase activity showed a significant increase in cerebellum of aluminum exposed rats. Most of the above parameters responded moderately to the individual treatment with CA and HEDTA, but significantly reduced blood and brain aluminum burden. However, more pronounced beneficial effects on some of the above described parameters were observed when CA and HEDTA were administered concomitantly. Blood and brain aluminum concentration however, showed no further decline on combined treatment over the individual effect with HEDTA or CA. We conclude that in order to achieve an optimum effect of chelation, combined administration of CA and HEDTA might be preferred. However, further work is needed before a final recommendation could be made.     

昆虫肠道蛋白酶作用下δ内毒素的毒性肽及毒力特异性的变化*

本文报道杀斜纹夜娥(Prodenia lirura)δ-内毒素和杀鞘翅目昆虫δ-内毒素经昆虫肠液蛋白酶及胰蛋白酶作用后,其毒性肽及毒力特异性的变化。 以Sephadex G75柱层析提纯的130kD原毒素,经斜纹夜蛾幼虫肠液蛋白酶作用后,产生的70kD与75kD抗蛋白酶多肽(PRP)对斜纹夜蛾和家蚕皆有毒;经家蚕幼虫肠液蛋白酶作用后,产生的62kD与65kD的PRP失去对斜纹夜蛾的毒性,仅对家蚕有毒;经胰蛋白酶作用后产生的65kD与68kD的PRP,其毒力特性与经家蚕肠液蛋白酶作用后的相似。斜纹夜蛾肠液蛋白酶作用后产生的PRP,可进一步被家蚕肠液蛋白酶或胰蛋白酶降解为63-65kD的PRP,此种多肽对斜纹夜蛾无毒,对家蚕有毒。杀鞘翅目昆虫的原毒素不能为胰蛋白酶和粘虫肠液所完全降解。证明不同种类昆虫的肠道蛋白酶对占-内毒素蛋白质的作用位点不同,δ-内毒素对宿主昆虫的毒力特异性与其肠道蛋白酶的特性密切相关。     

Crystallization of coupling factor 1 (CF1) from spinach chloroplast.

Spinach chloroplast coupling factor (CF₁) was crystal-lized at 20°C from 0.05 M TRIS-PO₄, containing 4 mM ATP, 15mM KCl, 1.0 mM EDTA and 1.80 M (NH₄)₂SO₄, at pH 7.8. Some unit cell parameters were determined by electron microscopy and by X-ray diffraction. The cube shaped crystals have a tetragonal lattice, a = b = 135 Å, c = 280 Å with eight molecules per unit cell; possible space group P422 or P4₂2₁2, hence half a molecule in the asymmetric unit. Crystals grown at pH 7.5 in the absence of ATP have an orthorhombic lattice, a = 125 Å, b = 145 Å, c = 169 Å (C222₁), eight molecules per unit cell.     

The role of 4,5-dioxovaleric acid in porphyrin and vitamin B12 formation by clostridia

4,5-Dioxovalerate, which has been proposed as an intermediate in the newly discovered so-called C₅ pathway that leads from L-glutamate to δ-aminolevulinate, strongly inhibits uroporphyrin formation from δ-aminolevulinate in cells of $\text{Clostridium}\text{tetanomorphum}$ and in cell-free extracts of this organism, in spite of the presence of L-alanine: 4,5-dioxovalerate aminotransferase (aminolevulinate aminotransferase, EC 2.6.1.43). The interference by 4,5-dioxovalerate with porphyrin formation is due to strong inhibition of δ-aminolevulinate dehydratase (EC 4.2.1.24). Since 4,5-dioxovalerate hence effectively prevents the operation of the reaction sequence from L-glutamate to porphyrin, it is concluded that 4,5-dioxovalerate does not function as a physiological δ-aminolevulinate precursor.     

Chloroplast culture VIII a new effect of kinetin in enhancing the synthesis and accumulation of protochlorophyllide invitro

By pretreating etiolated cucumber cotyledons with kinetin in the dark, it was observed that the plastids isolated from such tissues were 400% more active in the conversion of δ-aminolevulinic acid into protochlorophyllide, than plastids prepared from water-treated controls. The experimental evidence is consistent with the hypothesis that (a) the kinetin dark-pretreatment of the etiolated tissue, uncouples the joint biosynthesis of prothylakoids and protochlorophyll and results in the accumulation of excess prothylakoid membranes poorly supplied with protochlorophyllide (b) upon isolation of the plastids and incubation with δ-aminolevulinic acid, the latter is very rapidly converted into membrane-bound protochlorophyllide.     

Differential functions of PKC-delta and PKC-zeta in cisplatin response of normal and transformed thyroid cells

We investigated the effects of cisplatin (cisPt) in normal PC Cl3 and in transformed and tumourigenic PC E1Araf cells. cisPt cytotoxicity was higher in PC Cl3 than in PC E1Araf cells. In both cell lines, cisPt provoked the ERK1/2 phosphorylation; this was unaltered by G?6976, a conventional PKC inhibitor, whilst it was blocked by low doses (0.1 microM) or high doses (10 microM) of GF109203X, an inhibitor of all PKC isozymes, in PC Cl3 and in PC E1Araf cells, respectively. In PC E1Araf, the cisPt-provoked ERK phosphorylation was also blocked by the use of a myristoylated PKC-zeta pseudosubstrate peptide. Conversely, in PC Cl3 the cisPt-provoked ERK phosphorylation was blocked by the use of rottlerin, a PKC-delta inhibitor. Results show that cisPt activates both PKC (the -delta and the -zeta isozymes in PC Cl3 and in PC E1Araf cells, respectively) and ERK in association with prolonged survival of thyroid cell lines.     

Catabolism of 6-aminolaevulinic acid in etiolated Euglena gracilis

[4-¹⁴C]-δ-Aminolaevulinic acid (δ-ALA) was employed to study the specificity of δ-ALA incorporation into chlorophyll in etiolated Euglena gracilis. Degradation of chlorophyll a to acetate and propionate, obtained from ring B, indicated that, although some of the radioactivity was incorporated, it did not result from incorporation of intact δ-ALA.     

Eurotiaceae Trovate in Risaia

Abstract

Studying the mycological flora of paddy and grass-fields we had the opportunity to isolate some species of Eutotiaceae which are of particular interest. Three species were not yet known in Italy and only one was found in a soil of mountain pasture. The last one is also common in paddy fields. One of the isolated species was a Pseudoeurotium van Beyma, ascomycete with asci containing 16 ascospores, not identifiable with the systematically related species described as Pseudoeurotium indicum (Chattopadhyay and Das Gupta) Chattopadhyay from « oryzetis » in India and Pseudoeurotium multisporum (Saito and Minoura) Stolk in Japan. Some distinct morphological differences exist between our isolates and P. multisporum; but they differ only faintly from P. indicum: that is our isolates have pedicellate asci. Therefore we suggest the new variety: Pseudoeurotium indicum (Chattopadhyay and Das Gupta) Chattopadhyay var. orizetorum nobis. Three species of the genus Emericellopsis van Beyma: E. minima Stolk, E. terricola van Beyma and E. humicola (Cain) Cain were also isolated. The first one from the paddy-fields and the last two, not yet described in Italy, from grass fields. The possible relationship between E. minima Stolk arid E. pusilla Mathur, Sukapure and Thirumalachar has been discussed.