Cystine Uptake and Glutathione Level in Fetal Brain Cells in Primary Culture and in Suspension

Abstract— The glutathione level and the factors affecting this level were investigated in fetal rat brain cells in a primary culture. Early in the culture, the glutathione level of the brain cells decreased, but after 5 h it began to increase. This increase was not observed in a cystine-free medium and was prevented by excess glutamate. Cystine was taken up in freshly isolated brain cell suspensions, and its rate increased during the culture. The cystine uptake was mediated by a Na⁺-independent, glutamate-sensitive route previously found in various types of cells and designated as system x⁻_c. The uptake of cystine is a crucial factor in maintaining the glutathione level of the cells under culture, because it provides cysteine for the cells for glutathione synthesis. Cysteine was undetectable in the medium before the culture, but it appeared, though at a very low level, when the brain cells were cultured there. The source of this cysteine was the cystine in the medium. Presumably the decrease in the glutathione level of the cells in the early stage of the culture resulted from the fact that the medium did not contain cysteine. The enhancement of the cystine uptake during culture may constitute a protective mechanism against the oxidative stress to which the cultured cells are exposed. Regulation of the glutathione level in fetal brain cells in vivo by the transport of cystine and cysteine is discussed.     

Colony specific morphological caste differences in an Old World, swarm-founding polistine, Ropalidia romandi (Hymenoptera: Vespidae)

Morphological differences between queens and workers in an Australian swarm-founding paper wasp, Ropalidia romandi , were examined, providing detailed information on the nature of morphological caste differences in swarm-founding Ropalidia for the first time. Queens and workers differed in overall size and had divergent shapes, but differences were not always significant. In colonies in which queen–worker dimorphism was distinct, while the head width of queens was nearly the same as that of workers, queens were significantly larger in measures on the metasoma than workers. Plotting of metasomal variables against the mesosomal length and analysis of covariance showed that queen–worker morphological differentiation in R. romandi cannot be explained by simple linear allometric growth, suggesting that the two castes diverge in their growth parameters in the preadult stage. The pattern of morphological caste difference varied among colonies or was colony specific; there was no clear correlation between caste differences and colony cycle progression. These results suggest that morphological caste differences in R. romandi are determined genetically or by interactions between genetic background and intrinsic factors and/or external conditions.     

Red light causes a reduction in IAA levels at the apical tip by inhibiting de novo biosynthesis from tryptophan in maize coleoptiles

When maize coleoptiles were unilaterally exposed to red light (7.9 μmol m⁻²s⁻¹ for 5 min), 3 h after treatment IAA levels in coleoptiles decreased in all regions, from top to basal, with levels about 60% of dark controls. Localized irradiation in the 5 mm top zone was sufficient to cause the same extent of IAA reduction in the tips to that in the tips of whole irradiated shoots. When coleoptiles were treated with N-1-naphthylphthalamic acid (NPA), an accumulation of IAA in the tip and a decrease of diffusible IAA from tips were simultaneously detected. IAA accumulation in red-light treated coleoptiles by NPA was much lower than that of dark controls. NPA treatment did not affect the content of conjugated IAA in either dark or light treated coleoptile tips. When ¹³C₁₁ ¹⁵N₂-tryptophan (Trp) was applied to the top of coleoptiles, substantial amounts of stable isotope were incorporated into free IAA in dark and red-light treated coleoptile tips. The ratio of incorporation was slightly lower in red-light treated coleoptile tips than that in dark controls. The label could not be detected in conjugated IAA. The rate of basipetal transport of IAA was about 10 mm h⁻¹ and the velocity was not affected by red light. These results strongly suggest that red light does not affect the rates of conversion of free IAA to the conjugate form or of the basipetal transport, but just reduces the IAA level in the tips, probably inhibited by IAA biosynthesis from Trp in this region.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .     

Studies on Organophosphorus Insecticides

Various insecticides belonging to the O-alkyl-O-(subst. phenyl) phenylphosphonothioate family were prepared and their biological activities studied. Besides O-ethyl-O-(4-nitrophenyl) phenylphosphonothioate, which is utilized in these days, another compound, O-ethyl-O-(4-cyanophenyl) phenylphosphonothioate, proved to have excellent properties as an insecticide.

In our previous paper, it was reported that O,O-dimethyl-O-(4-cyanophenyl) phosphorothioate (S-4084) has only a low toxicity towards mammals though possessing a high activity towards rice stem borers. In this paper, the chemical and biological properties of S-4084 and its phenylphosphonothioate derivative (S-4087) will be reported in detail. It should especially be noted that S-4084 had a higher activity towards rice stem borers than S-4087 by topical or pot test in a room or in a green-house, but by field test S-4087 was stronger than S-4084.     

Action spectra for polarotropism and phototropism in protonemata of the fern Adiantum capillus-veneris

The action spectrum for polarotropism was determined, using the Okazaki large spectrograph, by brief irradiation with light between 260 nm and 850 nm in single-celled protonemata of the fern Adiantum capillus-veneris L., which had been cultured for 6 days in red light and then in the dark for 15 h. The action spectrum had a peak at around 680 nm. This effect was nullified by subsequent irradiaton with far-red light, and typical red/far-red reversibility was observed, indicating the involvement of phytochrome. Polarized ultraviolet or blue light had no effect on the direction of apical growth. The action spectrum for phototropism was also determined in the red light region by means of brief microbeam irradiation of a flank of the subapical region of the protonema. This spectrum showed a peak at 662 nm which was consistent with the absorption peak of phytochrome, but not with the peak of the action spectrum for polarotropism.     

Tissue Binding Patterns and In Vitro Effects of <Emphasis Type="Italic">Campylobacter jejuni</Emphasis> DNA-Binding Protein from Starved Cells

Campylobacter jejuni (C. jejuni) is frequently associated with axonal Guillain-Barré syndrome (GBS). We reported that C. jejuni DNA-binding protein from starved cells (C-Dps) binds to and damages myelinated nerves in vivo. We studied the binding patterns of C-Dps to nervous tissues and its in vitro effects on neural cells. Immunohistochemically, C-Dps labeled the nodes of Ranvier, the outermost parts of internodal myelin and the basement membrane in the peripheral nerves, and neurons and myelin in the central nervous tissues. Its binding was blocked by sulfatide. C-Dps bound to the cell surfaces of nerve growth factor (NGF)-treated PC12 cells leading to dose-dependent LDH release, which was inhibited by either heat-denaturation of C-Dps or coincubation with an anti-C-Dps mAb. However, its binding to the surfaces of cultured NSC34 cells, S16 cells, or dorsal root ganglion cells, did not induce cytotoxicity. These findings suggest a possible involvement of C-Dps in C. jejuni-related GBS.     

Purification and Some Properties of the Endo α-1,4 Polygalactosaminidase from Pseudomonas sp.

The endo α-1,4 polygalactosaminidase from Pseudomonas sp. 881 was purified from the culture nitrate by ethanol precipitation and sequential column chromatographies on CM-Sephadex C-25, Sephadex G-50 and Phenyl-Sepharose CL-4B. The purified enzyme was electrophoretically homogeneous and its molecular weight and isoelectric point were 31,000 and 6.7, respectively. The optimum pH and temperature for hydrolysis of polygalactosamine were 5.0 and 55°C, respectively. The enzyme was stable up to 45°C for 15min and from pH 4.0 to 7.6 at 37°C for 1 hr.

The Km value was 0.05% α-1,4 polygalactosamine and the V was 0.154μmol reducing sugar (galactosamine)/min/μg protein. This polygalactosaminidase was inhibited by Sn²⁺ , Fe²⁺ , Fe³⁺ , Hg²⁺, Cu²⁺ ions and SDS. The enzyme did not hydrolyze oligo galactosamines (n < tetramer) or N-acetyl-polygalactosamines. It acted only on oligo galactosamine (n > trimer) and polygalactosamine endogeneously so far tested.     

Identification of multiple quantitative trait loci affecting the size and shape of the mandible in mice

A quantitative trait locus (QTL) analysis was performed on the size and shape of the mandible in F₂ mice between KK-A ^y and C57BL/6 J strains and the effect of the A ^y allele on the morphology of the mandible was analyzed. A total of 13 measurements were taken on each right mandible. By means of discriminant and canonical discriminant analyses, KK-A ^y males and KK males were exactly discriminated from each other. In contrast to its known effects on body weight, the A ^y allele reduced the overall size of the mandible. QTL analysis of the 13 measurements and on three principal components extracted from these measurements identified multiple QTLs. When F₂ a/a and F₂ A ^y /a were analyzed separately, 11 significant main-effect QTLs were identified in F₂ a/a, whereas only two QTLs were identified in F₂ A ^y /a. Although four significant interactions were identified, all were in F₂ a/a. The A ^y allele thus made the difference in the size and shape of the mandible between strains obscure. Among mandible QTLs, those on Chrs 5 (Mssq6 and Mssq7) and 15 (Mssq14) were important. Mssq6 had an effect on the height of the posterior mandible. Mssq7 had an effect on mandible length. Mssq14 had an effect on the height of the anterior and posterior mandible. Mssq7 and Mssq14 also had an effect on the overall size. Thus, mandible QTLs have distinct and characteristic sites of action. Therefore, mandible morphology will be determined largely by the combination of these QTLs.