New Plant Growth Retardants

Phosphatidylcholine hydroperoxide was assayed with phospholipase A₂ and glutathione peroxidase, based on fluorometry with N(9-acridinyl)maleimide. The hydroperoxide was poorly reduced by glutathione peroxidase, and was converted by phospholipase A₂ into reactable forms of glutathione peroxidase. A linear relationship was found between hydroperoxides assayed by the enzymatic and chemical methods in the range from 0.05 to 5.0 nmol with 0.5 to 1.5 mg of the sample. The hydroperoxides of fatty acids, triacylglycerol and phosphatidylcholine were assayed in their mixtures and in commercial lecithins.     

Mice with Alopecia,Osteoporosis, and Systemic Amyloidosis Due to Mutation in Zdhhc13, a Gene Coding for Palmitoyl Acyltransferase

Protein palmitoylation has emerged as an important mechanism for regulating protein trafficking, stability, and protein–protein interactions; however, its relevance to disease processes is not clear. Using a genome-wide, phenotype driven N-ethyl-N-nitrosourea–mediated mutagenesis screen, we identified mice with failure to thrive, shortened life span, skin and hair abnormalities including alopecia, severe osteoporosis, and systemic amyloidosis (both AA and AL amyloids depositions). Whole-genome homozygosity mapping with 295 SNP markers and fine mapping with an additional 50 SNPs localized the disease gene to chromosome 7 between 53.9 and 56.3 Mb. A nonsense mutation (c.1273A>T) was located in exon 12 of the Zdhhc13 gene (Zinc finger, DHHC domain containing 13), a gene coding for palmitoyl transferase. The mutation predicted a truncated protein (R425X), and real-time PCR showed markedly reduced Zdhhc13 mRNA. A second gene trap allele of Zdhhc13 has the same phenotypes, suggesting that this is a loss of function allele. This is the first report that palmitoyl transferase deficiency causes a severe phenotype, and it establishes a direct link between protein palmitoylation and regulation of diverse physiologic functions where its absence can result in profound disease pathology. This mouse model can be used to investigate mechanisms where improper palmitoylation leads to disease processes and to understand molecular mechanisms underlying human alopecia, osteoporosis, and amyloidosis and many other neurodegenerative diseases caused by protein misfolding and amyloidosis.     

Some regulation profiles of ornithine transcarbamylase synthesis in vitro.

The regulation profiles of OTCase (argF, argI) synthesis in vitro were investigated by using the in vitro system described in the accompanying paper. Addition of 2.6 mM arginine, crude repressor and partially purified repressor to the in vitro system demonstrated that lambdadargF-DNA-directed OTCase-FFF synthesis is more sensitive to the repressor than lambdapargI-DNA-directed OTCase-III synthesis. The effects of some low-molecular substances on FFF and III syntheses were investigated; guanosine 3'-diphosphate 5'-diphosphate (ppGpp) stimulated both syntheses while cAMP and guanosine 5'-tetraphosphate (Gpppp) were not effective on III synthesis and were slightly inhibitory for FFF synthesis. The substances had no effect on the maturation of the enzyme or on the activity of the enzyme, FFF or III, synthesized. We suggest that argF- and argI-genes are regulated in a slightly different fashion and that the operator-promotor regions are not completely identical for these two genes.     

Degradation of 17β-Estradiol by a Gram-Negative Bacterium Isolated from Activated Sludge in a Sewage Treatment Plant in Tokyo, Japan

A 17β-estradiol (E2)-degrading bacterium was isolated from activated sludge in a sewage treatment plant in Tokyo, Japan. The isolate was suggested to be a new Novosphingobium species. Gas chromatography-mass spectrometry and ¹H nuclear magnetic resonance analyses of the metabolites of E2 degradation suggested that no toxic products accumulated in the culture medium.     

Two ultrastructurally distinct types of transporting tissues,the branchiostegal and the gill epithelia,in an estuarine tanaid,Sinelobus stanfordi (Crustacea,Peracarida)

Summary On each lateral side of the cephalothorax segments, the adult Sinelobus stanfordi has a branchial chamber which contains an elongated bag-shaped gill and is covered by a thick branchiostegite. The ultrastructural study revealed that the inner surface of the branchiostegite is composed of a transporting-type epithelium which is morphologically distinct from the gill epithelium. Both epithelia are covered by extremely thin (about 80 nm) cuticle layers, suggesting high permeability to gases, ions, and water. In contrast, the outer surface of the branchiostegite consists of ordinary epithelium covered by a very thick cuticle layer in common with the body surface. The inner branchiostegal epithelium (4–10 m thick) has a shallow (about 1 m deep) apical infolding system of the cell membrane (AIS) and an extensive three-dimensional tubular network (about 120 nm in diameter) which is formed by the invagination of the basolateral cell membrane (TNB). The TNB is associated with slender mitochondria and occupies the majority of the cytoplasmic area of the epithelial cell. The gill epithelium, on the other hand, is about 10 m thick and characterized by an abundance of oval mitochondria, well-developed (4–5 m deep) AIS, and a smooth basal cell membrane lacking any infoldings. These morphological features indicate that not only the gill epithelia, but also the inner branchiostegal epithelia, are involved in the ion-transporting processes. The ultrastructural differences between these two kinds of epithelia also suggest their different roles in the osmoregulation of this animal, since it inhabits estuaries which are subject to extreme changes in salinity.     

Streptomyces subtilisin inhibitor-like proteins are distributed widely in streptomycetes.

Streptomyces subtilisin inhibitor-like proteins were found to be distributed widely in streptomycetes by using the combination of the convenient, newly developed plate assay system and an established liquid culture assay. Almost all the strains formerly categorized as Streptoverticillium species produced proteins that exhibited inhibitory activity against both subtilisin BPN' and trypsin. N-terminal regions of three purified proteins showed high structural similarity to those of other previously reported SIL inhibitors.     

Comparison of the ability of lactate dehydrogenase-elevating virus and its virion RNA to infect murine leukemia virus-infected or -uninfected cell lines.

Lactate dehydrogenase-elevating virus (LDV) has a strict species specificity. Cells or cell lines other than a particular subset of mouse primary macrophages which can support LDV replication in vitro have not been identified. LDV induces neurological disorders in old C58 or AKR strains, in which the involvement of multiple copies of the endogenous N-tropic murine leukemia virus (MuLV) genome and the Fv-1 locus of the mouse has been implicated. Our previous studies have demonstrated that LDV could infect and replicate in cell lines of the mouse or other species in vitro when they were infected with MuLV. The significance of and the precise mechanism underlying this phenomenon, however, remain unclear. We demonstrated in this study the efficient infection and replication of the virus in vitro by inoculation of its RNA mixed with liposome. No significant difference either in the efficiency of RNA transfection or in the ability to support its replication was observed among the various species' cell lines examined. In addition, by RNA transfection the virus replicated with equal efficiency in MuLV-infected and -uninfected cells or in macrophages derived from mice irrespective of their age. In contrast, the pattern of the infection by virus particles was quite different; LDV replication was observed only in macrophages (particularly from newborn mice) and MuLV-infected cells. By using various LDV isolates, it was demonstrated that the capability of replication between neurovirulent, LDV type C, and the other avirulent strains was almost the same in mouse cell lines when their RNA was introduced into the cells. Higher infectivity of LDV-C to MuLV-infected cells may be due to its efficient incorporation of the particles into MuLV-infected cells.     

Differences in pectic polysaccharides between carrot embryogenic and non-embryogenic calli

Summary Cell walls and media were obtained from three kinds of carrot cell culture, namely, embryogenic callus (EC), non-embryogenic callus (NC) and somatic embryos (SE), and analyzed for their sugar content and sugar composition by electrophoresis and gas chromatography. EC formed large cell clusters while NC formed small clusters. Observations under the light microscope revealed that the intercellular contacts in NC were much more limited than those in EC. The analysis of pectic polysaccharides revealed that the level of neutral sugars was higher than that of acidic sugars in EC, while the opposite was true in NC. Gaschromatographic analysis of neutral sugars in pectic fractions revealed that EC and SE were rich in arabinose, while NC was rich in galactose. On the basis of these results, we discuss the possible involvement of neutral sugars, and of arabinose and galactose in particular, in pectic polysaccharides in intercellular contacts.Abbreviations EC embryogenic callus - NC non-embryogenic callus - SE somatic embryo - MS Murashige and Skoog - PAS periodic acid-Schiff s reagent