伪狂犬病病毒Ea株糖蛋白gD基因的表达及基因免疫

构建了2个利用人类巨细胞病毒（HCMV）的启动子启动表达伪狂犬病病毒Ea株糖蛋白gD基因的真核表达质粒pCIDI和pcDDI，体外转染BHK－21细胞，用间接免疫荧光法检测，证实糖蛋白gD在细胞中得到表达。用表达质粒pCIDI和pcDDI作为核酸疫苗免疫BALB/c小鼠，ELISA检测小鼠血清中抗伪狂犬病病毒的抗体，结果其滴度为1：128－1：1512。初步证实，用gD基因作为核酸疫苗免疫动物，可以激活机体的免疫应答反应。     

The carboxyterminal processing protease of D1 protein: expression, purification and enzymology of the recombinant and native spinach proteins

The carboxyterminal processing protease of D1 protein (CtpA) is predicted to be an excellent target for a general broad-spectrum herbicide. The gene for spinach CtpA has been expressed in Escherichia coli. The expressed protein that was found mainly in inclusion bodies has been purified and refolded on a nickel-chelate column. Active recombinant CtpA was recovered. Two assays for CtpA activity were developed, a medium-throughput HPLC assay using a fluorescent substrate and a high-throughput assay based on fluorescence polarization capable of application in a high-throughput 96-well plate format. This high-throughput assay was developed to screen chemistry for CtpA inhibitors. Native spinach CtpA was partially purified and the native and recombinant enzymes were compared kinetically for their K(m) and V(max) values using different peptide substrates. Native CtpA partially purified from spinach was shown to have similar kinetic properties to recombinant CtpA. Antibodies developed against the recombinant protein were used to estimate the in planta abundance of the native enzyme in spinach. Since only a small proportion of the recombinant protein is refolded during isolation and it appears that only a small proportion of this enzyme is active, size-exclusion chromatography and light scattering experiments were performed on rCtpA in order to gain insight into its structure and the reasons why most of the protein is not active. The use of rCtpA to screen for herbicidal compounds and the more general question of how good a herbicide target the enzyme is are discussed.     

Serum Levels of 25-Hydroxycholecalciferol and 1,25-Dihydroxycholecalciferol in Dogs with Hypercalcaemia

1,25-Dihydroxycholecalciferol (1,25-(OH)2-D3) and 25-hydroxycholecalciferol (25-OH-D3) were measured among dogs with hypercalcaemia (total serum calcium > 3.01 mmol/L) due to various causes. All values were compared to those of healthy control dogs. Serum 1,25-(OH)]2-D3 was measured by a radioimmunoassay test and serum 25-OH-D3 was measured by a protein binding assay. 1,25-(OH)2-D3 ranged from 26 to 332 pmol/L (median 110.0) in dogs with lymphoma (n = 12); from 61 to 398 pmol/L (median 248.0) in dogs with primary hyperparathyreoidism (n = 5); from 28 to 310 pmol/L (median 88.5) in dogs with chronic renal failure (n = 10); and from 60 to 239 pmol/L (median 157.5) in control dogs (n = 24). There was no significant difference in 1,25-(OH)2-D3 among dogs with different causes of hypercalcaemia. 25-OH-D3 ranged from 64 to 291 nmol/L (median 101.5) in dogs with lymphoma; from 66 to 298 nmol/L (median 91.0) in dogs with primary hyperparathyreoidism; from 35 to 184 nmol/L (median 67.0) in dogs with chronic renal failure; and from 48 to 350 nmol/L (median 306.5) in control dogs. 25-OH-D3 was significantly lower in dogs with lymphoma, primary hyperparathyroidism and chronic renal failure than in control dogs. 1,25-(OH)2-D3 and 25-OH-D3 are not predictable in dogs with hypercalcaemia.     

Reversal of the inhibition of photosynthesis by herbicides affecting hydroxyphenylpyruvate dioxygenase by plastoquinone and tocopheryl derivatives in Chlamydomonas reinhardtii

Isoxaflutole or pyrazolate inhibition of tocopherol and plastoquinone biosynthesis in the green alga Chlamydomonas reinhardtii Dang leads to the inactivation of photosystem II and the degradation of its reaction centre D1 protein when exposed to strong light. Cell-permeable short-chain derivatives of plastoquinone and tocopherol were tested in the reversal. Addition of decyl-plastoquinone reverses herbicide-induced inhibition of photosynthesis and inactivation of photosystem II in short-time (1 h) exposure of the algae to high light. In high light longer than 1 h, decyl-plastoquinone alone loses effectiveness, but a synthetic permeable tocopheryl derivative retards the inhibitory effects on photosystem II and on the degradation of the D1 protein. This indicates that tocopherol deficiency induced by the herbicides makes a major contribution to their secondary mode of action in high light stress.     

辣椒钴60γ辐射突变体的筛选及利用研究

1994年用3个剂量(5.16、7.74、10.32 C/kg)的钴60γ射线处理辣椒品种兰州大羊角干种子,从辐射剂量为5.16 C/kg的处理中经过连续10 a 选择,选育出性状表现稳定的新突变系C14.以突变系C14为母本,自交系0430为父本配制杂交组合2006D2,熟性早,果实羊角形,果面有皱褶,果长26.0 cm,肉厚0.25 cm,单果质量50～55 g,果色绿,味辣,果实商品性好,品质优良,抗病毒病,耐疫病.前期产量和总产量分别为28 718.55 kg/hm2、64 351.95 kg/hm2,比对照陇椒2号分别增产2.2%、5.9%.适宜于北方保护地和露地栽培.     

Incidence of positive heartworm antibody and antigen tests at IDEXX Laboratories: trends and potential impact on feline heartworm awareness and prevention

Data from the IDEXX Laboratories Reference Laboratory Network were retrospectively examined for feline heartworm testing trends in testing frequency, geographic bias, and prevalence for the years 2000--2006. Examination of the data supports the commonly held view that veterinarians do not embrace heartworm disease testing or prevention in cats to the same degree they do in dogs. Despite significant awareness and adoption of heartworm testing and prevention in dogs, we hypothesized that heartworm testing rates are lower for cats than for dogs despite a significant prevalence of feline infection. This is important because a perceived low rate of infection in cats is likely to manifest in a low adoption of testing and prevention. In reality, the overall feline heartworm antigen-positive rate is significant--on average 0.9% over the period studied--and in some regions was estimated to be as high as 4.6%. This compares with an average canine heartworm prevalence rate of 1.2%, a feline leukemia virus prevalence of 1.9%, and a feline immunodeficiency prevalence of 1.0%. Based on the low rate of testing and these prevalence rates, practitioners are routinely missing cases of adult feline heartworm infections and the recently defined heartworm-associated respiratory disease (H.A.R.D). Increased antigen testing would result in detection of a significant number of positive cases. In addition, this population of infected cats would represent the "tip of the iceberg" relative to the greater number of cats that have early infection or are at risk for infection.     

Vitamin D receptor amounts across different segments of the gastrointestinal tract in Brown Swiss and Holstein Frisean cows of different age

During different stages of lactation, different requirements of calcium have to be met depending on the milk amount. Vitamin D receptors (VDR) regulate calcium homeostasis by increasing the entry of Ca into blood from bone stores and dietary sources. The purpose of this study was to investigate if age and breed of cows influence VDR amounts across different segments of the gastrointestinal tract. Thirty-six cows were used (18 Brown Swiss, 18 Holstein Friesan, both > 5.5 years or < 4.5 years). Tissue specimens of the intestines were collected from the cows. Formaldehyde-fixed and microwave-treated paraffin sections were used for VDR immunohistochemistry employing a biotinylated monoclonal rat antibody and streptavidin peroxidase technique. The results showed that nuclei and cytoplasm of enterocytes stained positively for VDRs. Strongest immunoreactions were observed in intermediate and basal glandular cells. No significant differences were observed between the different groups. Vitamin D receptors immunoreactivities were prominent in duodenal mucosa, lower in jejunum and in colon, decreased further in ileum and were lowest in caecum. Decreases in number of positively marked cells and staining intensities resulted in reduced immunoreactions. The results of this study indicate that VDR are highly expressed at the site of maximal intestinal calcium absorption. No significant influence of age and breed was observed. The animals used were not in a negative Ca balance. The cows were all in the stage of late or mid lactation. During these periods, the Ca requirements are low and the diets are high in Ca concentration; and the animals are adapted to these circumstances. Passive absorption in adult animals seems to dominate when Ca intake is adequate or high. The active absorption may play a considerably more significant role during the peripartal period, when Ca homeostatic mechanisms are challenged because of tremendous Ca demand at the initiation of lactation.     

Preventing species invasion: A role for integrative taxonomy?

Integrative taxonomy, a multi‐disciplinary approach adding modern techniques to traditional morphology‐based methods (e.g. molecular and morphological criteria), can play an important role in bioinvasion research to identify introduced taxa, discover pathways of introduction and inform authorities to control and prevent future introductions. The present study is the first on introduced populations of Callosciurus, Asiatic tree squirrels, known as potentially invasive species in Europe (Italy, Belgium and France). We combined molecular (mitochondrial DNA markers: CoxI, D‐loop) and morphometric analysis on skulls, comparing them to the widest morphological and molecular datasets ever assembled for Callosciurus. Squirrels collected in Italy and Belgium share the same haplotypes and skull characteristics, but are conspicuously different from the French population in Antibes. Genetic data revealed close similarity between French squirrels and Pallas's squirrels, Callosciurus erythraeus, from Taiwan, China. Italian and Belgian squirrels formed an independent taxonomic lineage in genetic analyses, whose taxonomic rank needs further investigation. The morphological and morphometric characteristics of these 2 populations are, however, similar to known specimens assigned to Callosciurus erythraeus. These results may indicate a common origin for the populations found in Belgium and Italy. In contrast, French specimens suggest an independent introduction event of squirrels originating from Asia.     

Expression of APOB,ADFP and FATP1 and their correlation with fat deposition in Yunnan’s top six famous chicken breeds

ABSTRACT

1. Adipose differentiation related protein (ADFP), fatty acid transport protein 1 (FATP1) and apolipoprotein B (APOB) are suspected to play an important role in determining intramuscular fat and in overall meat quality.

2. Yunnan’s top six famous chicken breeds (the Daweishan Mini, Yanjin Black-bone, Chahua, Wuding, Wuliangshan Black-bone and Piao chicken) are known for the high quality of their meat, but little is known about their expression of these three genes.

3. The present study aimed to examine the ADFP, FATP1 and APOB genes in different tissues of these six breeds at different development stages. The subcutaneous fat from the back midline and front, abdominal fat, liver and muscle tissue was sampled at 28, 49, 70, 91 and 112 days. The expression of ADFP, FATP1 and APOB was measured by real-time PCR.

4. The results showed that the expression of the three genes differed depending on age, tissue types and breeds. However, the expression of the three genes correlated with fat traits. In conclusion, the expression of the ADFP, FATP1 and APOB genes is associated with the fat traits of Yunnan’s top six chicken breeds. These results could help with molecular marker screening and marker-assisted breeding to improve the quality of poultry for meat production.