Expression of the tobacco β-1,3-glucanase gene, PR-2d, following induction of SAR with Peronospora tabacina

Systemic acquired resistance (SAR) is induced following inoculation of Peronospora tabacina sporangia into the stems of Nicotiana tabacum plants highly susceptible to the pathogen. Previous results have shown that accumulation of acidic β-1,3-glucanases (PR-2's) following induction of SAR by P. tabacina may contribute to resistance to P. tabacina. We showed that up-regulation of the PR-2 gene, PR-2d, following stem inoculation with P. tabacina, is associated with SAR. Studies using plants transformed with GUS constructs containing the full length promoter from PR-2d or promoter deletions, provided evidence that a previously characterized regulatory element that is involved in response to salicylic acid (SA), may be involved in regulation of PR-2d following induction of SAR with P. tabacina. This work provides evidence that regulation of PR-2 genes during P. tabacina-induced SAR may be similar to regulation of these genes during infection of N-gene tobacco by TMV or following exogenous application of SA, and provides further support for the role of SA in regulation of genes during P. tabacina-induced SAR.     

DCAB对荷斯坦奶牛泌乳性能、血液酸碱平衡及血清矿物质含量的影响

试验选用荷斯坦泌乳早期奶牛15头,采用3X3复拉丁方试验设计,研究日粮DCAB水平分别为+367、+514、+663mmol/kgDM对奶牛生产性能、血液酸碱平衡和血清矿物质含量的影响。试验结果,不同DCAB水平日粮对奶牛奶产量差异不显著(P>0.05),高DCAB水平日粮可增加奶牛血液标准HCO-3含量(P<0.05),不同DCAB水平对奶牛各种血清常量矿物质含量没有影响(P>0.05)。试验表明,日粮DCAB水平超过+514mmol/kgDM可对奶牛奶产量产生不利影响。     

HBV/HAV复合抗原基因在CHO细胞中的表达

为探讨HBV／HAV复合疫苗的可行性，利用DNA重组技术将HBsAg与HAW复合多表位抗原基因VPX进行融合，插入到真核表达质粒pVAX1多克隆位点，构建成核酸表达疫苗pVAX1／SVPX，将其转染CHO细胞。转染细胞培养48h后，进行RTPCR、Western-blot、ELISA分析，结果表明HBV／HAV复合抗原基因SVPX能够在CHO细胞内转录、表达，融合蛋白具有HBV和HAV抗原的特性。     

0～3周龄北京鸭氨基酸理想模式的研究

依据扣除法原理，根据现有氨基酸需要量标准和目前的研究结果规划出一种氨基酸模式，依次将模式中赖氨酸、蛋氨酸、色氨酸、苏氨酸、异亮氨酸扣除20％，而非扣除的氨基酸量保持不变，来确定扣除某一种氨基酸后对育雏期北京鸭生产性能和血液生化指标的影响，确定出理想的氨基酸比例。试验共设6个处理组，为6种氨基酸模式，每个处理设4个重复。结果表明：北京鸭育雏期氨基酸理想模式为赖氨酸：蛋氨酸：色氨酸：苏氨酸：异亮氨酸=100：42：22：38：48。     

核酸疫苗研究进展(下)

Sato等报道，在质粒载体上引入大肠杆菌Amp^ 抗性基因(内含有6核苷酸片段的ISS免疫增强序列)，真皮免疫小鼠后ISS增强CTL和Th1细胞的免疫反应。有人提出利用含有病毒免疫原性基因的基因大片段进行基因免疫，以弥补单基因免疫活性的不足。将协同因子如IL-2、IL-12、TCA等的表达质粒与抗原表达质粒混合注射可使Th1型细胞免疫增强，而GMcsf能明显加强体液免疫，     

Distribution of alpha‐neoendorphin,ACTH (18–39) and beta‐endorphin (1–27) in the alpaca brainstem

Using an immunocytochemical technique, we have studied in the alpaca brainstem the distribution of immunoreactive structures containing prodynorphin (alpha‐neoendorphin)‐ and pro‐opiomelanocortin (adrenocorticotrophin hormone (18–39) (ACTH), beta‐endorphin (1–27))‐derived peptides. No peptidergic‐immunoreactive cell body was observed. Immunoreactive fibres were widely distributed, although in most of the brainstem nuclei the density of the peptidergic fibres was low or very low. In general, the distribution of the immunoreactive fibres containing the peptides studied was very similar. A close anatomical relationship occurred among the fibres containing alpha‐neoendorphin, ACTH or beta‐endorphin (1–27), suggesting a functional interaction among the three peptides in many of the brainstem nuclei. The number of fibres belonging to the prodynorphin system was higher than that of the pro‐opiomelanocortin system. A moderate/low density of immunoreactive fibres was observed in 65.11% (for alpha‐neoendorphin (1–27)), 18.18% (for ACTH) and 13.95% (for beta‐endorphin) of the brainstem nuclei/tracts. In the alpaca brainstem, a high density of immunoreactive fibres was not observed. The neuroanatomical distribution of the immunoreactive fibres suggests that the peptides studied are involved in auditory, motor, gastric, feeding, vigilance, stress, respiratory and cardiovascular mechanisms, taste response, sleep‐waking cycle and the control of pain transmission.     

Genetic risk factors for osteochondrosis in various horse breeds

Osteochondrosis (OC) is an injury to cartilage canals with a following necrosis in the growth cartilage, from there it can develop to osteochondrosis dissecans (OCD). Due to its high impact in the equine industry, new insights into predisposing factors and potential high‐risk genetic variants are warranted. This article reviews advancements in quantitative and molecular genetics in refining estimation of genetic parameters and identifying predisposing genetic loci. Heritabilities were highest for hock OC with estimates at 0.29–0.46 in Hanoverian warmblood and Norwegian trotters, whereas in Thoroughbreds only very low genetic variation seemed to be present in hock OC lesions. Whole genome scans using the Illumina Equine SNP50 or SNP70 Beadchip were performed in Thoroughbred, Standardbred, French and Norwegian trotter, Hanoverian and Dutch warmblood. Validation studies in Spanish Purebred and Hanoverian warmblood horses corroborated OC risk loci on ECA 3, 14, 27 and 29. Particularly, a strong association with hock‐OCD was found for a single nucleotide polymorphism (SNP) on horse chromosome (ECA) 3 upstream to the LCORL gene. Gene expression and microRNA analyses may be helpful to understand pathophysiological processes in equine OC and to connect OCD‐associated genomic regions with potential candidate genes. Furthermore progress in elucidating the underlying genetic variants and pathophysiological changes in OC may be expected from whole genome DNA and RNA next‐generation sequencing studies.     

Canine GM2‐Gangliosidosis Sandhoff Disease Associated with a 3‐Base Pair Deletion in the HEXB Gene

Background

GM2‐gangliosidosis is a fatal neurodegenerative lysosomal storage disease (LSD) caused by deficiency of either β‐hexosaminidase A (Hex‐A) and β‐hexosaminidase B (Hex‐B) together, or the GM2 activator protein. Clinical signs can be variable and are not pathognomonic for the specific, causal deficiency.

Objectives

To characterize the phenotype and genotype of GM2‐gangliosidosis disease in an affected dog.

Animals

One affected Shiba Inu and a clinically healthy dog.

Methods

Clinical and neurologic evaluation, brain magnetic resonance imaging (MRI), assays of lysosomal enzyme activities, and sequencing of all coding regions of HEXA, HEXB, and GM2A genes.

Results

A 14‐month‐old, female Shiba Inu presented with clinical signs resembling GM2‐gangliosidosis in humans and GM1‐gangliosidosis in the Shiba Inu. Magnetic resonance imaging (MRI) of the dog's brain indicated neurodegenerative disease, and evaluation of cerebrospinal fluid (CSF) identified storage granules in leukocytes. Lysosomal enzyme assays of plasma and leukocytes showed deficiencies of Hex‐A and Hex‐B activities in both tissues. Genetic analysis identified a homozygous, 3‐base pair deletion in the HEXB gene (c.618‐620delCCT).

Conclusions and Clinical Importance

Clinical, biochemical, and molecular features are characterized in a Shiba Inu with GM2‐gangliosidosis. The deletion of 3 adjacent base pairs in HEXB predicts the loss of a leucine residue at amino acid position 207 (p.Leu207del) supporting the hypothesis that GM2‐gangliosidosis seen in this dog is the Sandhoff type. Because GM1‐gangliosidosis also exists in this breed with almost identical clinical signs, genetic testing for both GM1‐ and GM2‐gangliosidosis should be considered to make a definitive diagnosis.