The embryonic vertebrate heart tube is a dynamic suction pump

The embryonic vertebrate heart begins pumping blood long before the development of discernable chambers and valves. At these early stages, the heart tube has been described as a peristaltic pump. Recent advances in confocal laser scanning microscopy and four-dimensional visualization have warranted another look at early cardiac structure and function. We examined the movement of cells in the embryonic zebrafish heart tube and the flow of blood through the heart and obtained results that contradict peristalsis as a pumping mechanism in the embryonic heart. We propose a more likely explanation of early cardiac dynamics in which the pumping action results from suction due to elastic wave propagation in the heart tube.     

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.     

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.     

Ellagic acid inhibits oxidized low-density lipoprotein (OxLDL)-induced metalloproteinase (MMP) expression by modulating the protein kinase C-α/extracellular signal-regulated kinase/peroxisome proliferator-activated receptor γ/nuclear factor-κB (PKC-α/ERK/PPAR-γ/NF-κB) signaling pathway in endothelial cells

Previous studies have shown that vascular endothelium-derived matrix metalloproteinases (MMPs) contribute to the destabilization of atherosclerotic plaques, a key event triggering acute myocardial infarction. In addition, studies have reported that the PKC-MEK-PPARγ signaling pathway is involved in oxidized low-density lipoprotein (oxLDL)-induced expression of MMPs. Ellagic acid, a phenolic compound found in fruits and nuts, has potent antioxidant, anti-inflammatory, and anticancerous properties. However, the molecular mechanisms underlying its antiatherogenic effects remain to be clarified. This study aimed to assess whether the effects of ellagic acid on the fibrotic markers MMP-1 and MMP-3 are modulated by the PKC-ERK-PPAR-γ signaling pathway in human umbilical vein endothelial cells (HUVECs) that have been exposed to oxLDL. It was found that ellagic acid significantly inhibited oxLDL-induced expressions of MMP-1 and MMP-3. Pretreatment with ellagic acid and DPI, a well-known ROS inhibitor, attenuated the oxLDL-induced expression and activity of PKC-α. In addition, ellagic acid as well as pharmacological inhibitors of ROS, calcium, and PKC strongly suppressed the oxLDL-induced phosphorylation of extracellular signal-regulated kinase (ERK) and NF-κB activation. Moreover, ellagic acid ameliorated the oxLDL-induced suppression of PPAR-γ expression. In conclusion, the data suggest that ellagic acid elicits its protective effects by modulating the PKC-α/ERK/PPAR-γ/NF-κB pathway, resulting in the suppression of ROS generation and, ultimately, inhibition of MMP-1 and MMP-3 expression in HUVECs exposed to oxLDL.     

Expression of recombinant phytase of Bacillus subtilis E20 in Escherichia coli HMS 174 and improving the growth performance of white shrimp,Litopenaeus vannamei,juveniles by using phytase‐pretreated soybean meal‐containing diet

To produce a thermostable and neutral phytase (phy) of Bacillus subtilis E20 in Escherichia coli HMS174 and evaluate its efficiency in improving growth performance. The phy C of B. subtilis E20 was expressed in E. coli HMS 174, and then the 42‐kDa recombinant phy C was purified by Ni‐NAT and analysed by SDS–PAGE. The recombinant phy C had optimal ranges of pH of 6 ~ 7 and temperature of 50 ~ 60 °C. A thermostability analysis showed that the enzyme is a thermostable phytase, and around 33% of residual activity was detected after being incubated at 90 ~ 100 °C for 10 min. The recombinant phy C‐pretreated soybean meal for feed preparation improved white shrimp, Litopenaeus vannamei, growth and feed efficiency. Overall, the neutral and thermostable phy C is suitable for aquafeed, and it is able to improve the nutritional utilization, resulting in enhanced shrimp growth and reduced feed costs.     

利用DNA分子标志鉴别台湾茶树品种及评估种原之遗传歧异性

为评估台湾茶树种原之遗传歧异性。本研究由100条ISSR引子申筛选出12条可产生多型性条带明显的引子．这些引子共可产生67个的多型性条带。依据每一种原之分子标志数据进行UPGMA法分群分析结果。可将台湾133个茶树种原区分成六大群，包括油茶群、赤芽山茶群、野生茶树群、大叶变种与小叶变种混合群、大叶、小叶及大叶、小叶杂交种混合群及小叶变种群。而主成分向量分析的结果与利用群聚分析得到的亲缘关系树形固结果相符合。台湾茶树种原高比例的遗传歧异度是由台湾的野生茶树所贡献．部分重要栽培种间的相似性仍极高。为了探讨制茶过程封分子级品种鉴定之影响及DNA分子标志应用于咸茶品种鉴定之可行性，本研究分析不同发酵程度的茶类。在制茶过程申封DNA质量之影响。试验结果显示高温杀菁过程严重造成咸茶DNA的降解。利用各种类别咸茶与新鲜茶叶（封照）所抽取之DNA样品进行PCR扩增反应．结果发现分子量小于1,000bp的ISSR DNA条带表现较稳定。     

Acceleration of post‐mortem changes in Tsaiya duck (Anas platyrhynchos) breast muscle by lactic acid marination

1. The effect of lactic acid marination at 5°C on post mortem changes in breast muscle pectoralis major of spent layer Tsaiya duck was studied.

2. Myofibrils were prepared from 0.1 M and 0.2 M lactic acid marinated muscle and control (non‐marinated samples) sampled at 0, 1, 3, 7 and 14 d post mortem.

3. Changes in myofibril fragmentation index (MFI), myofibrillar proteins and Z‐line structure were examined.

4. Marination of duck breast muscle in lactic acid at 5°C enhanced fragmentation of myofibrils and degradation of myofibrillar proteins and Z‐line structure as compared to control samples.

5. In summary, lactic acid marination at 5°C can accelerate the post mortem degradation of myofibrils in Tsaiya duck breast muscle.     

Size distribution approaches for monitoring and conservation of coastal Cymodocea habitats

• 1. Cymodocea nodosa's leaf length distribution was studied as an easily measurable indicator to monitor and conserve Macedonian, North Aegean, Greek coastal habitats.
• 2. Three Cymodocea meadows off the eastern Kavala Gulf coast (Nea Karvali, Erateino, Agiasma), with that of Nea Karvali close to an industrial area being the most degraded, were sampled during the seagrass main growing season in July 2004. Two further meadows, one pristine to less degraded (Brasidas, Gulf of Kavala) and one degraded (Biamyl, Inner Thessaloniki Gulf), were sampled as benchmarks in July 2005. The results were evaluated using Gaussian fit curves, and non‐parametric and nested parametric ANOVA on a hierarchy of spatial scales: area (tens of metres), site (hundreds of metres) and meadow (kilometres).
• 3. Frequency (%) distribution of leaf length values and CymoSkew index variation were best associated with anthropogenic stress. Frequency (%) distribution of adult and intermediate photosynthetic leaf length values revealed a unimodal distribution possible to be fitted, at least at pristine to less degraded meadows, by normal distribution (R²>0.5).
• 4. Statistically significant variation was estimated for CymoSkew index, a quantitative expression of leaf length asymmetry, on the meadow scale (P<0.001). Biamyl (3.82) and Nea Karvali (3.64) were indicated as heavily degraded meadows, Erateino (2.93) as a degraded meadow, Agiasma (2.18) as a meadow with the first signs of degradation, and Brasidas (1.68) as a pristine to less degraded meadow. These results in combination with other meadow specific biotic parameters were used to suggest a preliminary angiosperm ‘Ecological Status Classes’ classification scheme useful for the implementation of WFD in the north Aegean Sea.
• 5. The CymoSkew index seems to respond to lower levels of stress than is needed for other more conservative plant modules and therefore, could be regarded as an early warning indicator of Cymodocea habitat degradation. Copyright © 2009 John Wiley & Sons, Ltd.