Polymorphisms in the canine glutamate transporter-1 gene: identification and variation among five dog breeds

Excitatory amino acid transporters (EAATs) are important for terminating glutamatergic neurotransmission and protect central nervous system (CNS) neurons from glutamatergic excitotoxicity. We selected these genes as targets that may relate to canine behavioral traits. After screening four EAAT genes (glutamate transporter-1; GLT-1, excitatory amino acid transporter 4; EAAT4, excitatory amino acid carrier; EAAC1, glutamate/aspartate transporter; GLAST) for single nucleotide polymorphisms (SNPs), we identified two silent SNPs (C129T and T471C) in the GLT-1 gene. We genotyped 193 dogs of 5 breeds and found significant variation among breeds in these two SNPs in GLT-1. The C129T polymorphism was not observed in Malteses and Miniature Schnauzers. These results suggest that polymorphisms in the GLT-1 gene may be useful markers for examining how the genetic background relates to the behavioral traits of dogs.     

Isolation and characterization of some antioxidative compounds from the rhizomes of smaller galanga (Alpinia officinarum Hance)

Antioxidative compounds were isolated from the methanol extract of fresh rhizome of smaller galanga (Alpinia officinarum Hance). Seven phenylpropanoids (1-7) were finally obtained by reversed-phase HPLC, and their structures were elucidated by MS and NMR analyses. They comprised the two known compounds, (E)-p-coumaryl alcohol gamma-O-methyl ether (1) and (E)-p-coumaryl alcohol (6), and the five novel compounds, stereoisomers of (4E)-1,5-bis(4-hydroxyphenyl)-1-methoxy-2-(methoxymethyl)-4-pentene (2a and 2b), stereoisomers of (4E)-1,5-bis(4-hydroxyphenyl)-1-ethoxy-2-(methoxymethyl)-4-pentene (3a and 3b), (4E)-1,5-bis(4-hydroxyphenyl)-1-[(2E)-3-(4-acetoxyphenyl)-2-propenoxy]-2-(methoxymethyl)-4-pentene (4), (4E)-1,5-bis(4-hydroxyphenyl)-2-(methoxymethyl)-4-penten-1-ol (5), and (4E)-1,5-bis(4-hydroxyphenyl)-2-(hydroxymethyl)-4-penten-1-ol (7). Compounds 1-7 were detected for the first time as constituents of galanga rhizomes and exhibited antioxidative activities against the autoxidation of methyl linoleate in bulk phase.     

Acceleration of follicular development by administration of vascular endothelial growth factor in cycling female rats

To address the role of follicular angiogenesis in the determination of ovulatory follicles and the effects of different vascular endothelial growth factor (VEGF) isoforms on follicular angiogenesis and development, mature female rats were treated with an angiogenic inhibitor (TNP-470), and also with VEGF 120 or 164 at different dosages (0.4, 0.8, 4.0 or 8.0 microg/kg body weight) for 3 days during the estrous cycle. Ovarian follicular angiogenesis, the population of large follicles and ovulation were examined. VEGF 120 (0.8 microg/kg) and 164 (8.0 microg/kg) treatments stimulated follicular angiogenesis in the theca interna layer, while TNP-470 treatment showed severe depression of follicular angiogenesis, and completely inhibited ovulation. After administration of VEGF 120 or 164, the number of healthy preovulatory follicles and ovulated oocytes increased significantly, concomitantly with a decrease in the number of atretic preovulatory follicles. The oocytes ovulated had normal fertilizability and developed to term with the same litter size as in the control rats. Our findings suggest that follicular angiogenesis may be a determinant of follicular development during the periovulatory phase, and that VEGF isoforms may play different important roles in regulating follicular angiogenesis.     

Influence of feeding regime on timing of parturition in beef cattle and the relationship of vaginal temperature to parturition

The timing of parturition was recorded for a total of 56 beef cattle (Japanese Black × Holstein Friesian) on different dietary treatments. The rate of calving during daylight hours in cows night‐fed (18.00 hours) with a roughage diet was significantly higher than that in cows night‐fed with a high concentrate diet (79.2% vs 38.5%, P < 0.05). Subsequently, the vaginal temperature (VT) of these cows was analyzed using a cosinor method. When the feeding schedule was changed from twice daily (08.30 and 15.30 hours) to night feeding, the periodicity, the acrophase and the bathyphase, which were the parameters of the cosine curve, were unstable from the first day of night feeding until after day 6 (P < 0.05). Prior to parturition, the midline‐estimating statistic of rhythm (MESOR) and the amplitude for the cows that were fed a high‐roughage diet at night and that calved at night‐time were lower and larger, respectively, than that for the other treatments (P < 0.01). Based on these results, the time of parturition in most of the beef cows was influenced by feeding time and diet composition. Those cows that calved at night‐time in spite of night feeding had lower vaginal temperatures.     

Identification of oyster-derived hypotensive peptide acting as angiotensin-I-converting enzyme inhibitor

Angiotensin-I-converting enzyme (ACE) plays a crucial role in the crisis of hypertension. Some peptides that originate from protease hydrolysates are known to suppress ACE activity in vitro and in vivo. Here, we investigated whether trypsin hydrolysate of oyster Crassostrea gigas showed hypotensive activity and ACE inhibition. The hydrolysate significantly suppressed systolic blood pressure and ACE activity in spontaneously hypertensive rats following a one-shot oral administration and a long-term feeding experiment lasting 9 weeks. Each hydrolysate from oyster tissue showed ACE inhibitory activity, indicating the hypotensive effect was due to synergism. One potent ACE inhibitory peptide, Asp-Leu-Thr-Asp-Tyr, was identified from the hydrolysate of the striate muscle, and the peptide exhibited hypotensive activity in vivo. Protease digestion analysis suggested that Asp-Tyr could be the real effector of this penta-peptide in vivo.     

Investigation of the prevalence of neurologic equine herpes virus type 1 (EHV-1) in a 23-year retrospective analysis (1984–2007)

A single nucleotide polymorphism in the equine herpesvirus 1 (EHV-1) DNA polymerase gene (ORF30 A₂₂₅₄ to G) has been associated with clinical signs of equine herpes myeloencephalopathy (EHM). The purpose of our study was to determine the odds ratio for this genetic marker and EHM using a panel of field isolates from North America collected over the past twenty-three years. EHV-1 isolates cultured at the Cornell University Animal Health Diagnostic Laboratory from 1984 to 2007 were retrieved along with their clinical histories. DNA was extracted from these EHV-1 cultures and allelic discrimination was performed using real-time PCR. The results were confirmed by sequencing of the target region in ORF30. PCR and sequencing were in 100% agreement and showed that 19 out of the 176 isolates had the ORF30 G₂₂₅₄ allele (11%), of which16 were EHM cases and 3 respiratory or abortion cases. The odds of having neurologic disease with the ORF30 G₂₂₅₄ genotype were computed as 162 times greater than those with the opposite allele ORF30 A₂₂₅₄ (95% confidence interval: 35–742). Despite this strong statistical significance, 24% (5/21) of horses with neurologic disease in our study population harbored the “non-neurologic” form of the allele (ORF30 A₂₂₅₄), suggesting that other factors may also contribute to the onset of EHM.     

The profile of urinary lipid metabolites in healthy dogs

Polyunsaturated fatty acids, including arachidonic acid (AA), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA), are converted to hundreds of lipid mediators by cyclooxygenases (COX), lipoxygenases (LOX), and cytochrome P450 (CYP), or through non-enzymatic processes, and they reflect inflammatory states of the body. We comprehensively analyzed lipid metabolites in dog urine using a liquid chromatograph-mass spectrometry (LC-MS/MS) to describe their metabolic characteristics. We detected 31 AA-derived metabolites, four EPA-derived metabolites, and a DHA-derived metabolite in all urine samples. Among AA-derived metabolites, 15, 5, 3, and 8 were generated by COX, LOX, CYP, and non-enzymatic oxidation respectively. This study will be the first step to use profiles of urinary lipid metabolites for better understanding and diagnosis of canine diseases.     

Interaction between the helicase domain of the tobacco mosaic virus replicase and a tobacco arginine decarboxylase

In a yeast two-hybrid screening test for tobacco proteins that interact with TMV replicase using the helicase (H) domain as bait, a cDNA clone was selected that encodes a polyamine biosynthetic enzyme, arginine decarboxylase (ADC). In yeast cells, the C-terminal internal region of ADC interacted with the H domain. This observation was confirmed in vitro by far-Western blotting. Inhibition of the binding between the H domain and the IRnHEL (I region and N-terminus of helicase domain) region by ADC using a yeast three-hybrid assay suggested possible interference of the heterodimerization of 126K and 183K by ADC.The nucleotide sequence data of pADCF reported in this study is available in the DDBJ/EMBL/GenBank databases under accession number AB110952     

不同含水量下工具材料切向土壤黏附特性（摘选）

为了提高越野车辆性能和建筑、农业领域机械工作效率,就需要了解土壤黏附机理。该试验旨在找到不同工具材料的土壤切向黏附与含水率之间的关系,探讨土壤切向黏附对土壤剪切力学性能的影响。试验对橡胶（RUB）、钢（SS400）和树脂（PEEK）3种材料进行了直剪试验。结果表明,土壤和材料之间的切向黏附随着土壤含水量的增加而增加至最大值后降低。土壤切向黏附最大值的排序是钢（SS400）,其次是橡胶（RUB）,最后是树脂（PEEK）。结果还表明,3种材料道具与测试土壤间的外摩擦角随着土壤含水率的增加先降低后增加,当发生剪切的时候,随着剪切的发生,土壤含水量导致切向黏附的变化进而直接影响剪切应力和外摩擦角。