Abnormal uterus with polycysts, accumulation of uterine prostaglandins, and reduced fertility in mice heterozygous for acyl-CoA synthetase 4 deficiency.

Arachidonate released by various stimuli is rapidly reesterified into membrane phospholipids initiated by acyl-CoA synthetase (ACS) and subsequent acyl-transfer reactions. ACS4 is an arachidonate-preferring enzyme abundant in steroidogenic tissues and postulated to modulate eicosanoid production. Female mice heterozygous for ACS4 deficiency become pregnant less frequently and produce small litters with extremely low transmission of the disrupted alleles. Striking morphological changes, including extremely enlarged uteri and lumina filled with numerous proliferative cysts of various sizes, were detected in ACS4+/- females. Furthermore, marked accumulation of prostaglandins was seen in the uterus of the heterozygous females. These results indicate that ACS4 modulates female fertility and uterine prostaglandin production.     

Elimination of artifactual immunogold labeling from secondary walls of woody stem sections

Preliminary in situ transmission electron microscopy immunogold localization of a 24-kDa dehydrin-like protein in red-osier dogwood (Cornus sericea L.) stem cross-sections was contaminated with extensive background labeling of secondary cell walls in both positive and negative control samples. Alterations in antibody dilution, buffer salt concentration and stringency of the washing solutions failed to eliminate background cell-wall labeling. A procedure was developed in which lyophilized cold-acclimated C. sericea xylem tissue was pulverized and boiled with sodium dodecyl sulfate (SDS)-protein extraction buffer to remove soluble proteins and to inactivate proteases. Wood powder, treated with SDS-protein extraction buffer, was used to pre-absorb chicken immune serum specific for a 24-kDa dehydrin-like protein prior to immunolocalization assays. Pre-incubation of primary antibodies did not compromise the recognition of the 24-kDa protein, and this technique effectively eliminated background cell-wall labeling.     

Evolutionarily Conserved Interaction between the Phosphoproteins and X Proteins of Bornaviruses from Different Vertebrate Species

Bornavirus, a non-segmented, negative-strand RNA viruses, is currently classified into several genetically distinct genotypes, such as Borna disease virus (BDV) and avian bornaviruses (ABVs). Recent studies revealed that bornavirus genotypes show unique sequence variability in the putative 5′ untranslated region (5′ UTR) of X/P mRNA, a bicistronic mRNA for the X protein and phosphoprotein (P). In this study, to understand the evolutionary relationship among the bornavirus genotypes, we investigated the functional interaction between the X and P proteins of four bornavirus genotypes, BDV, ABV genotype 4 and 5 and reptile bornavirus (RBV), the putative 5′ UTRs of which exhibit variation in the length. Immunofluorescence and immunoprecipitation analyses using mammalian and avian cell lines revealed that the X proteins of bornaviruses conserve the ability to facilitate the export of P from the nucleus to the cytoplasm via interaction with P. Furthermore, we showed that inter-genotypic interactions may occur between X and P among the genotypes, except for X of RBV. In addition, a BDV minireplicon assay demonstrated that the X and P proteins of ABVs, but not RBV, can affect the polymerase activity of BDV. This study demonstrates that bornaviruses may have conserved the fundamental function of a regulatory protein during their evolution, whereas RBV has evolved distinctly from the other bornavirus genotypes.     

Chemical analysis of the female sex pheromone in Palpita nigropunctalis (Lepidoptera: Crambidae)

The lilac pyralid, Palpita nigropunctalis Bremer (Lepidoptera: Crambidae), is a common pest of Oleaceae plants. A crude extract of the female sex pheromone glands was examined by gas chromatography-electroantennogram detection (GC-EAD) and GC coupled to a mass spectrometer (GC/MS). The GC-EAD analysis revealed three EAG-active components (I–III) in a ratio of 1:0.2:0.01 (I: II: III). GC/MS analysis successfully recorded the mass spectra of I and II. For I, ions at m/z 238 (M⁺) and 220 ([M-18]⁺) indicated the structure of a monoenyl aldehyde with a 16-carbon chain. For II, M⁺ was not detected, but ions at m/z 222 ([M-60]⁺) and 61 ([AcOH+1]⁺) suggested that II was a monoenyl acetate with a 16-carbon chain. Further GC/MS analysis of the extract treated with dimethyl disulfide revealed that the double bonds in both I and II are located at the same position of 11th-carbon. In addition, the pheromone extract was examined by GC/Fourier transform-infrared spectrophotometer (GC/FT-IR). An IR spectrum of I showed characteristic absorption at 1716 and 966?cm^?1, indicating a formyl group and E configuration of the double bond, respectively. In the case of II, absorption at 1745 and 968?cm^?1 indicated an ester carbonyl and E configuration, respectively. Taken together and by comparison with authentic standards, I and II were confirmed as (E)-11-hexadecenal and (E)-11-hexadecenyl acetate, respectively; while III was speculated as (E)-11-hexadecen-1-ol. The synthetic I, II and III all coincided well with those of the natural components in chemical data, and elicited strong electroantennographic activity in male P. nigropunctalis.     

Retroviral gene transfer for the assignment of Fanconi anemia (FA) patients to a FA complementation group

Fanconi anemia (FA) is an autosomal recessive disorder characterized by bone marrow failure, cancer susceptibility, and a variety of developmental defects. The disease is clinically heterogeneous; eight different complementation groups (FA A–H) and, thus, genetic loci have been discovered. Two genes, FAA and FAC, have been cloned. Disease-associated mutations have been detected and rapid mutation screening makes possible the assignment of patients without resorting to time-consuming cell fusion and complementation analysis. Amplification of specific cDNAs from RNA followed by direct or indirect sequence analysis is a standard method for mutation detection. During the course of such examinations of the FAC gene, we have noted that frequently only one of the expressed alleles is successfully amplified. This can lead to false assignment of patients to a complementation group. As we report here, such cases can be rapidly clarified by retroviral gene transfer and complementation analysis. Received: 30 July 1997 / Accepted: 13 October 1997     

Aggregation of Lepidostomatidae in small mesh size litter-bags: implication to the leaf litter decomposition process

Invertebrate colonization during leaf litter decomposition was studied at the 2nd order of Yanase River, Iruma city, Saitama, Japan from November 13, 2002 to May 20, 2003. Two different mesh sizes (1 and 5 mm) of litter-bags were used to evaluate the decomposition of leaf litter of Sakura (Prunus lannesiana), bags were placed equally in riffle (water flow velocity: 0.2–0.6 m s⁻¹) and pool (water flow velocity: 0.04–0.06 m s⁻¹). Mass loss and invertebrates in the litter-bags were monitored at interval between 1 and 3 weeks, and the invertebrates were classified based on their functional feeding group. Among the invertebrates found inside the litter-bags, the case-bearing shredder Lepidostomatidae was the most dominant invertebrates and they were the early colonizer that appeared about 3 months after the litter-bags immersion. In absence or low number of leaf-shredders, the decomposition rates in 1 and 5 mm litter mesh bags followed the exponential (or first-order) decay kinetic (R ²: 0.72–0.92). However, the presence of a large number of leaf-shredders in 1 mm litter-bags caused an acceleration of decomposition process; that even resulted faster mass loss than the loss from the 5 mm mesh bags placed in riffle area (0.030 day⁻¹ vs. 0.011 day⁻¹). Our results shows the importance of using different mesh sizes of litter-bags in decomposition study, which is applicable to the experiment in lotic or lentic ecosystem. Using smaller mesh size of litter-bags can provide information on how significant the effect of detritus feeders on the decomposition process, while the bigger mesh size can represent better the natural decomposition process when a large number detritus feeders is present in the smaller mesh size of litter-bags.