Identification of sex-specific polymorphic sequences in the goat amelogenin gene for embryo sexing

Amelogenin (AMEL) is a conserved gene located on the sex chromosomes of mammals. It is involved in the formation of enamel, which is the hard, white material that forms the protective outer layer of each tooth. In this study, we first cloned and determined the intron sequences of the goat AMELX and AMELY genes from female and male ear tissues. The polymorphic AMEL alleles were further analyzed by PCR-based RFLP and Southern blot hybridization analyses. Results showed that intron 5 nucleotide sequences of the goat AMELY gene contains multiple deletions/insertions and shares only 48.5% identity to intron 5 of the goat AMELX gene. Based on the polymorphic AMEL intron sequences, a set of sex-specific triplex primers was designed to PCR amplify a single fragment of 264 bp from the X chromosome of female goats and 2 fragments of 264 and 206 bp from the X and Y chromosomes, respectively, of male goats. An increased sensitivity for sex determination was reached with a single blastomere at the blastula stage isolated from goat embryos. A total of 43 goat embryos were used to estimate a 100% accuracy rate of this method confirmed by chromosomal karyotyping and live births. The embryo sexing technique has been successfully applied in different strains of goats including Alpine, Saanen, Nubian, and Taiwan goats.     

Precocene II metabolism: Comparative in vivo studies among several species of insects,and structure elucidation of two major metabolites

Several species of insects, exhibiting varying responsiveness to the juvenile hormone antagonist precocene II (6,7-dimethoxy-2,2-dimethylchromene), were challenged topically with a tritiated preparation of the title compound. Metabolism of [³H]precocene II was subsequently examined by withdrawing hemolymph samples from treated animals at appropriate time intervals and characterizing the extractable radiolabel chromatographically. Quantitative (or qualitative) differences observed between the respective metabolic profiles were found not correlative with specimen sensitivity to precocene. Production of two heretofore unreported metabolites, identified by spectral and chemical means as O-β-glucosides of 6- and 7-monodemethylated precocene II, was demonstrated in both sensitive and insensitive species. No evidence for the presence of a hemolymphborne, biologically effective “activated metabolite” produced in vivo by precocene-susceptible insects could be found. The latter finding may well argue for in situ bioactivation of precocene at the target tissue(s) by these sensitive insects.     

Inhibition of juvenile hormone biosynthesis in the tobacco hornworm, Manduca sexta, by a bisthiolcarbamate juvenoid and related compounds

Biosynthesis of juvenile hormone in the tobacco hornworm, Manduca sexta, is inhibited by the bisthiolcarbamate juvenoid N-ethyl-1,2-bis(isobutylthiolcarbamoyl)ethane both in vitro and in vivo. In vitro an extremely steep dose-response curve was obtained with an ID₅₀ value of 6 × 10^?6M. However, in vivo topical treatment with the compound resulted in mild JH antagonistic symptoms, suggesting rapid metabolism of the compound. In agreement with results from metabolic studies performed on plants and in mammals, sulfoxidation of the thiocarbamate S-(4-chlorobenzyl)N,N-diethylthiocarbamate resulted in an enhanced inhibitory effect on JH biosynthesis in vitro. This suggests that the corresponding thiocarbamate sulfoxides may act as intermediates in carbomylating critical thiol sites important in the terpenoid biosynthesis pathway. Furthermore, this study shows that these prototype compounds are interesting tools for further investigation of chemical inhibition of JH biosynthesis in insects.     

Molecular characterization of the CP gene and 3'UTR of Chilli veinal mottle virus from South and Southeast Asia

Twenty-four isolates of Chilli veinal mottle virus (ChiVMV) from China, India, Indonesia, Taiwan and Thailand were analysed to determine their genetic relatedness. Pathogenicity of virus isolates was confirmed by induction of systemic mosaic and/or necrotic ringspot symptoms on Capsicum annuum after mechanical inoculation. The 3' terminal sequences of the viral genomic RNA were determined. The coat protein (CP) coding regions ranged from 858 to 864 nucleotides and the 3' untranslated regions (3'UTR) from 275 to 289 nucleotides in length. All isolates had the inverted repeat sequence GUGGNNNCCAC in the 3'UTR. The DAG motif, conserved in aphid-transmitted potyviruses, was observed in all isolates. All 24 isolates were considered as belonging to ChiVMV because of their high CP amino acid and nucleotide identity (more than 94·8 and 89·5%, respectively) with the reported ChiVMV isolates including the pepper vein banding virus (PVBV), the chilli vein-banding mottle virus (CVbMV) and the CVbMV Chiengmai isolate (CVbMV-CM1). Based on phylogenetic analysis, ChiVMV isolates including all 24 isolates tested, PVBV, CVbMV and CVbMV-CM1 can be classified into three groups. In addition, a conserved region of 204 amino acids with more than 90·2% identity was identified in the C terminal of the CP gene of ChiVMV and Pepper veinal mottle virus (PVMV), and may explain the serological cross reaction between these two viruses. The conserved region may also provide useful information for developing transgenic resistance to both ChiVMV and PVMV.     

北京地区春玉米螟害产量损失估计方法及防治指标的商榷

根据1962—1964年北京地区12块春播玉米地心叶期卵和卵虫(第一代)成活率資料,并参考国内对螟害与玉米产量損失关系的研究成果,提出了春玉米上因玉米螟为害而造成的产量損夫估計方法和药剂防治的参考指标。心叶期卵的平均成活率及其标准誤为57.4±2.9％,卵块的脫落是死亡的主要原因。幼虫期的平均成活率及其标准誤为5.64±0.89％。这两个平均成活率的变異系数都比较小,相关分析表明,百株着卵量与成长幼虫数是相关的。故可职利用卵和幼虫的平均成活率由着卵密度来估計成长幼虫密度。心叶期卵和幼虫的合計平均成活率及其标准誤为3.4±0.51％。在95％可靠性时的置信区間为3.4±1.173％,卵块的平均粒数为31.7粒。若职单株平均一虫所造成的产量損失率为5％計,可用丁式来估計产量損失: 产量損失％=[(心叶期百株累計卵块数×31.7)×(0.034±0.012)]×0.05 在經济核算士,作者初步認为职損失率1.5％作为防治标准比較合适。以此推算,欲达到这一損失率,心叶期百株累計卵块应为28块(用平均成活率計算)至21块(用成活率上界計算)。一般可定为24块。根据心叶期百株累計卵块数与累計着卵株百分率或百株高峯卵块数之間存在的相关,百株累計24块卵,相当于累計着卵株率28％,或百株高峯卵块6块。在实践上就可以用它們作为心叶期的防治指标。     

Genetic relationships of Rhododendron (Ericaceae) in Taiwan based on the sequence of the internal transcribed spacer of ribosomal DNA

Summary

The genetic relationships of 20 Rhododendron species in Taiwan was determined based on the sequence of the internal transcribed spacer (ITS) region of ribosomal DNA. Sequences of the complete ITS region including ITS1, 5.8S rDNA, and ITS2, were obtained by direct sequencing of polymerase chain reaction (PCR)-amplified fragments. Gaultheria itoana was used as an outgroup. Aligned sequences of ITS1 and ITS2 from the 21 taxa resulted in 493 characters. According to the dendrogram, six main clusters were classified among the 20 species of the genus Rhododendron in Taiwan. Rhododendron oldhamii, R. nakaharai, R. taiwanalpinum, R. simsii, R. lasiostylum, R. rubropilosum, R. breviperulatum, R. kanehirai, and R. noriakianum were grouped with R. longiperulatum in cluster I. Rhododendron lamprophyllum was grouped with R. ovatum in cluster II. Rhododendron pseudochrysanthum, R. morri, R. hyperythrum, and R. rubropunctatum were grouped with R. formosanum in cluster III. In addition, R. mariesii, R. ellipticum and R. kawakamii formed three independent clusters. In this study, the findings based on ITS sequences are in agreement with the systematics of Rhododendron.     

Earthquake in a maze: compressional rupture branching during the 2012 M(w) 8.6 Sumatra earthquake

Seismological observations of the 2012 moment magnitude 8.6 Sumatra earthquake reveal unprecedented complexity of dynamic rupture. The surprisingly large magnitude results from the combination of deep extent, high stress drop, and rupture of multiple faults. Back-projection source imaging indicates that the rupture occurred on distinct planes in an orthogonal conjugate fault system, with relatively slow rupture speed. The east-southeast-west-northwest ruptures add a new dimension to the seismotectonics of the Wharton Basin, which was previously thought to be controlled by north-south strike-slip faulting. The rupture turned twice into the compressive quadrant, against the preferred branching direction predicted by dynamic Coulomb stress calculations. Orthogonal faulting and compressional branching indicate that rupture was controlled by a pressure-insensitive strength of the deep oceanic lithosphere.     

Dihydroaustrasulfone Alcohol Inhibits PDGF-Induced Proliferation and Migration of Human Aortic Smooth Muscle Cells through Inhibition of the Cell Cycle

Dihydroaustrasulfone alcohol is the synthetic precursor of austrasulfone, which is a marine natural product, isolated from the Taiwanese soft coral Cladiella australis. Dihydroaustrasulfone alcohol has anti-inflammatory, neuroprotective, antitumor and anti-atherogenic properties. Although dihydroaustrasulfone alcohol has been shown to inhibit neointima formation, its effect on human vascular smooth muscle cells (VSMCs) has not been elucidated. We examined the effects and the mechanisms of action of dihydroaustrasulfone alcohol on proliferation, migration and phenotypic modulation of human aortic smooth muscle cells (HASMCs). Dihydroaustrasulfone alcohol significantly inhibited proliferation, DNA synthesis and migration of HASMCs, without inducing cell death. Dihydroaustrasulfone alcohol also inhibited platelet-derived growth factor (PDGF)-induced expression of cyclin-dependent kinases (CDK) 2, CDK4, cyclin D1 and cyclin E. In addition, dihydroaustrasulfone alcohol inhibited PDGF-induced phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), whereas it had no effect on the phosphorylation of phosphatidylinositol 3-kinase (PI3K)/(Akt). Moreover, treatment with PD98059, a highly selective ERK inhibitor, blocked PDGF-induced upregulation of cyclin D1 and cyclin E and downregulation of p27^kip1. Furthermore, dihydroaustrasulfone alcohol also inhibits VSMC synthetic phenotype formation induced by PDGF. For in vivo studies, dihydroaustrasulfone alcohol decreased smooth muscle cell proliferation in a rat model of restenosis induced by balloon injury. Immunohistochemical staining showed that dihydroaustrasulfone alcohol noticeably decreased the expression of proliferating cell nuclear antigen (PCNA) and altered VSMC phenotype from a synthetic to contractile state. Our findings provide important insights into the mechanisms underlying the vasoprotective actions of dihydroaustrasulfone alcohol and suggest that it may be a useful therapeutic agent for the treatment of vascular occlusive disease.     

Carriage speed control of a cross-lapper system for nonwoven web quality

In a cross-lapper system, the carriage unit is used to make uniform web in appropriate width and thickness. Traditionally, this is a stable, sluggish system with large attenuation of the input in the steady state. In this paper, the modeling, stability, order reduction, dynamic analysis and controller design of a cross-lapper system for good nonwoven web quality will be presented to deal with this issue. A realizable controller is designed which can not only make the closed-loop system efficient for good tracking property but also can achieve the meaningful design objectives. The performance can be effectively shown from the computer simulation.