Structures of spider toxins: hydroxyindole-3-acetylpolyamines and a new generalized structure of type-E compounds obtained from the venom of the Joro spider, Nephila clavata

Facile structure determination of acylpolyamines, glutamatergic nerve blocker obtained from the venom of the Joro spider (Nephila clavata) was carried out with the use of micro-column LC/MS and high energy collision induced dissociation (CID) mass spectrometry. 6-hydroxyindole-3-acetyl was proposed previously as a putative partial structure, for the acyl moiety of hydroxyindole-type polyamines (NPTX-1 to -6). The NMR data obtained for NPTX-6, NPTX-687 and hydroxyindole-3-acetic acid which was released by acid hydrolysis of Nephila clavata crude venom extracts proved that the lipophilic head is the 4-hydroxyindole-3-acetic acid. Various hydroxyindole-3-acetyl polyamines were found in N. Clavata venom and characterized by mass spectrometry. As a result, type-E, a new class of generalized acylpolyamine structure was proposed in addition to the previously reported polyamine backbones type-A to -D.     

Evidence against a direct role for the induction of c-jun expression in the mediation of drug-induced apoptosis in human acute leukemia cells

Previous reports have demonstrated that a variety of anticancer drugs, e.g., 1-beta-D-arabinofuranosylcytosine (ara-C), mitoxantrone, etoposide, camptothecin, and cisplatin, induce the expression of c-jun oncogene in leukemic cells prior to producing internucleosomal DNA fragmentation and the morphological features of apoptosis. This has led to the impression that the induction of c-jun expression may be directly involved in the molecular signaling of the final common pathway of programmed cell death or apoptosis. In the present study, we examined the role of c-jun expression in three different settings of anticancer drug-induced apoptosis in human leukemic cells. First, exposure of human myeloid leukemia HL-60 cells to high-dose ara-C for 4 h produced internucleosomal DNA fragmentation preceded by c-jun induction. However, pretreatment of HL-60 cells with staurosporine, a protein kinase C inhibitor, repressed c-jun yet enhanced DNA fragmentation and apoptosis due to ara-C. Second, in human pre-B leukemia 697/BCL-2 cells which are transfected with the cDNA of the bcl-2 oncogene and overexpress p26BCL-2, although ara-C or mitoxantrone treatment caused greater c-jun induction than in the 697/neo cells, significantly reduced endonucleolytic DNA fragmentation and apoptosis was observed in 697/BCL-2 cells. Finally, taxol-induced internucleosomal DNA fragmentation and morphological features of apoptosis in HL-60 cells were not associated with the induction of c-jun expression. These lines of evidence indicate that the induction of c-jun expression may not have a direct role in the molecular signaling of anticancer drug-induced apoptosis, and that the anticancer drug-induced apoptosis can occur by a mechanism that does not involve the induction of c-jun expression.     

Fabrication of low-loss single-mode fibres

Low-loss single-mode fibres with GeO2-doped SiO2 cores have been made by the c.v.d. method and the loss spectrum from 0.6 to 1.8 ?m measured. The minimum loss of the fibre with suitably designed parameters, characterised by the refractive-index difference and the core diameter, was 0.75 dB/km at 1.05 ?m.     

Application of a novel polypropylene to the insulation of an electric power cable

Cross‐linked polyethylene (XLPE) has been widely adopted as insulating material for high‐voltage power cables up to 500 kV. Further improvement of electrical and thermal properties on insulating material is required in order to increase cable operation efficiency. Therefore, the development of novel insulating material possessing high thermal properties will be necessary. Recent progress of catalysis technology contributes to obtain new polymeric materials which may be applied to electrical insulation. The authors investigated the basic properties of newly developed stereoregular syndiotactic polypropylene (s‐PP) which is synthesized with homogeneous metallocene catalyst. Though recycling of cross‐linked polymers such as conventionally used XLPE may be difficult because of their poor heat deformation, the s‐PP which is not cross‐linked must be suitable for recycling. A series of experiments on its physical and electrical properties gave the following results.

• (1) s‐PP has sufficient flexibility compared with isotactic polypropylene (i‐PP ).
• (2) Both AC and lightning impulse breakdown strength of s‐PP in spite of no cross‐linking are superior to those of XLPE in the temperature range from 25 to 90 °C.
• (3) Degradation by copper of s‐PP is less than that of i‐PP.
• (4) s‐PP/VLDPE blend shows sufficient brittleness temperature for use.

These results suggested that s‐PP should serve as insulating material for power cables at higher‐temperature operation. © 2003 Wiley Periodicals, Inc. Electr Eng Jpn, 146(1): 18–26, 2004; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002//eej.10210     

Noncompetitive inhibition by L-cysteine and activation by L-glutamate of the iron-oxidizing activity of a mixotrophic iron-oxidizing bacterium strain OKM-9

A mesophilic, mixotrophic iron-oxidizing bacterium strain OKM-9 uses ferrous iron as a sole source of energy and L-glutamate as a sole source of cellular carbon. Uptake of L-glutamate into OKM-9 cells is absolutely dependent on ferrous iron oxidation. Thus, the Fe(2+)-dependent L-glutamate uptake system of strain OKM-9 is crucial for the bacterium to grow mixotrophically in iron medium with L-glutamate. The relationship between iron oxidation and L-glutamate transport activities was studied. Iron oxidase containing cytochrome a was purified 9-fold from the plasma membrane of OKM-9. A purified iron oxidase showed one rust-colored band following disc gel electrophoresis after incubation with Fe(2+). The Fe(2+)-dependent L-glutamate transport system was also purified 14.5-fold from the plasma membrane using the same purification steps as for iron oxidase. Fe(2+)-dependent L-glutamate and L-cysteine uptake activities of OKM-9 were 0.36 and 0.24 nmol/mg/min, respectively, when a concentration of 18 mM of these amino acids was used as a substrate. Both uptake activities were completely inhibited by potassium cyanide (KCN), suggesting that cytochrome a in the iron oxidase is involved in the transport process. The iron-oxidizing activity of strain OKM-9 was activated 1.7-fold by 80 mM L-glutamate. In contrast, the activity was noncompetitively inhibited by L-cysteine. The Michaelis constant of iron oxidase for Fe(2+) was 12.6 mM and the inhibition constant for L-cysteine was 41.6 mM. A marked inhibition of iron oxidase by 50 mM L-cysteine was completely reversed by the addition of 60 mM L-glutamate. The results suggest the possibility that iron oxidase has a binding site for L-cysteine and the cysteine first bound to the iron oxidase was replaced by the added L-glutamate.     

Efficient construction of cDNA microarrays utilizing normalized cDNA libraries of Arabidopsis thaliana

We have demonstrated the utility of normalization for efficient cDNA microarray preparation using Arabidopsis as a model. Nonredundant cDNAs including 5722 species were efficiently collected from random 7914 expressed sequence tags (ESTs) in four normalized cDNA libraries. The prepared microarrays were successfully used to monitor gene expression. These methodologies should be applicable to the study of other species in plant biotechnology.     

Simulation of confined and enhanced optical near fields for a long narrow aperture in a pyramidal structure on a thick metallic screen

A new type of long narrow aperture in a pyramidal structure on a thick metallic screen is proposed, and optical wave scattering by this structure is simulated. This aperture structure provides high emission intensity and small spot size simultaneously through excitation of the surface plasmon polaritons on the sidewalls of the pyramidal structure. Scattering of optical waves by this structure in the thick metallic screen is solved numerically with a volume integral equation by generalized conjugate residual iteration and fast Fourier transformation. The basic characteristics of the near-field intensities of the aperture are investigated in detail.