Use of electrospray mass spectrometry for mass determination of grape (Vitis vinifera) juice pathogenesis-related proteins: a potential tool for varietal differentiation

Methods based on liquid chromatography-mass spectrometry (LC-MS) and protein trap mass spectrometry (trap-MS) were developed to determine the complement of pathogenesis-related (PR) proteins in grape juice. Trap-MS was superior to LC-MS in terms of simplicity, rapidity, and sensitivity. Proteins with a wide range of masses (13--33 kDa) were found in the juices of 19 different varieties of grape (Vitis vinifera) and were identified as mostly PR-5 type (thaumatin-like) and PR-3 type (chitinases) proteins. Although the PR proteins in juices of grapes are highly conserved, small consistent differences in molecular masses were noted when otherwise identical proteins were compared from different varieties. These differences persisted through different harvest years and in fruits grown in different Australian locations. With the definition of four different masses for PR-5 proteins (range = 21,239--21,272 Da) and nine different masses of PR-3 proteins (range = 25,330--25,631 Da) and using statistical analysis, the methods developed could be used for varietal differentiation of grapes grown in several South Australian locations on the basis of the PR protein composition of the juice. It remains to be seen whether this technology can be extended to grapes grown worldwide and to wine and other fruit-derived products to assist with label integrity to the benefit of consumers.     

Marked induction of IL-6, haptoglobin and IFNgamma following experimental BRSV infection in young calves

Bovine respiratory syncytial virus (BRSV) has been identified worldwide as an important pathogen associated with acute respiratory disease in calves. An infection model has been developed reflecting accurately the clinical course and the development of pathological signs during a natural BRSV-infection. In the experiments described in the present study, calves were infected at 13-21 weeks of age and reinfected 14 weeks later. Blood samples from the entire infection period were analysed for acute phase protein (haptoglobin) by ELISA and for expression (mRNA level in peripheral blood mononuclear cells) of the cytokines interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-6 (IL-6) and interferon-gamma (IFNgamma) by quantitative real-time reverse transcribed polymerase chain reaction (RT-PCR). IFNgamma, interleukin-6 and haptoglobin were markedly induced together with development of clinical signs in response to the first infection with BRSV. The IFNgamma response was biphasic, with an early peak at day 1-3 post infection (p.i.) and a later increase between day 5 and 8 p.i. Reinfection also resulted in an induction of IFNgamma, but without induction of clinical signs, IL-6 and haptoglobin. These results indicate that early mediators connected with the innate responses are induced on a first encounter with the pathogen, but not on a second encounter (reinfection) where the adaptive immune system may act as the first line defence.     

转生长激素基因工程鲤鱼研究

约10000余尾注射牛和羊生长激素基因的鲤鱼已被生产出来，并从中筛选出生长速度快且整合了外源基因的个体143尾，也证明外源基因可传递给子代同时子代也具有快速生长的特征。建立了把外源基因直接注入受精卵的方法，确定了注入外源基因的最佳时期。     

Functional characterization of post-junctional adrenergic receptor subtypes in bovine intra-mammary arteries

We examined the functional role of adrenergic receptor subtypes (ARs) in bovine intra-mammary arteries (IMAs), 1.5–2.5 mm internal diameter. Norepinephrine (NE) and phenylephrine (PE) produced concentration-dependent increases in tone in segments maintained at a previously determined optimal basal tension in vitro . The sensitivity of the tissue to NE and PE, based on -log molar ED_50s was 6.87 ± 0.17 and 7.05 ± 0.35, respectively. In addition a Schild analysis yielded antagonist affinities for the receptor mediating contractile responses to NE (pA2 value) of 10.46 ± 0.85 for prazosin and 6.29 ± 0.18 for yohimbine. These data indicate a dominance of functional alpha 1 (α₁) over alpha 2 (α₂)-ARs in this tissue. Based on the inhibitory effects of chloroethylclonidine (CEC) on PE responses and the further reduction in sensitivity when nifedipine was added to the CEC, also in the presence of PE, we conclude that there is more than one α₁-AR subtype, with a predominant role for α_1B-ARs in phenylephrine responses. Stimulation of beta (β)-ARs, resulted in relatively small reductions in tone (the highest magnitude of response was 25.94 ± 6.46% of the papaverine maximum at 3×10⁻⁶ M isoproterenol); in addition, propranolol did not significantly alter tissue sensitivity to NE. Additional characterization of functional autonomic receptor populations in this circulatory bed will form a basis for future studies on circulatory dynamics in the mammary gland.     

Closantel缓释剂对实验感染肝片吸虫山羊机体自由基影响的研究

选用5只实验感染肝片吸虫山羊研究Closantel缓释剂对机体抗氧化功能的影响，实验用山羊每只一次性口服肝片吸虫囊蚴150个，感染后第15周随机将山羊分成对照组（n=2）和驱虫组（n=3），驱虫组每羊口服两颗Closantel缓释剂，每周定时采集颈静脉血液，检测血清抗氧化酶的变化，观察服用Closantel缓释剂对其的影响。结果表明投药后山羊血清中GSH－Px、CAT均有不同程度的升高，SOD变化不显著，而脂质过氧化产物MDA含量则下降，提示Closantel的缓释剂型对实验感染肝片吸虫山羊机体的抗氧化产生了影响，对机体有一定保护作用。     

Cardiac arrhythmias and electrolyte disturbances in colic horses

Background

Despite increased focus on cardiac arrhythmias in horses, the nature and prevalence is still poorly described. Case reports suggest that arrhythmias occurring secondary to systemic disease are seen more commonly in the clinic than arrhythmias caused by cardiac disease. The aim of this study was to investigate the prevalence of arrhythmias in colic horses referred for hospital treatment. Associations between electrolyte disturbances and arrhythmias were also investigated.The study population consisted of eight control horses and 22 referred colic horses. A Holter electrocardiography (ECG) was recorded during the first 24 hours of admission. The ECG’s were analysed by a software program followed by manual visual inspection. Arrhythmias registered included second degree atrioventricular (AV) blocks, supraventricular premature complexes (SVPCs), and ventricular premature complexes (VPCs). Blood was collected at admission and again between 12 and 24 hours after ECG was applied, and analysed for concentrations of potassium, sodium, ionised calcium, chloride, glucose, and L-lactate.

Results

Heart rate was 37.4 ± 3.7 bpm in the control group, and 51.6 ± 11.8 bpm, in the colic group, which was significantly different (P < 0.0001). AV blocks and SVPCs were found in both groups, however only colic horses showed VPCs. No significant difference between the two groups was found for AV blocks, SVPCs, and VPCs (P = 0.08 - 0.76). The mean levels of potassium, sodium, ionized calcium, and chloride were significantly lower in the colic group compared to the control group at admission. Mean levels of glucose and L-lactate were significantly elevated in the colic group (P < 0.05).

Conclusions

This study describes prevalence of cardiac arrhythmias and electrolytes concentrations in colic horses compared to healthy controls. Although we only observed VPCs in the colic horses, no significant differences between colic horses and controls were found. Despite the colic horses having electrolyte changes at admission no correlation was found between the electrolyte disturbances and cardiac arrhythmias. Although no clear conclusions can be drawn from the present study, the results indicate that relatively mild colic per se is not pro-arrhythmogenic, whereas severe colic probably are more likely to result in ventricular arrhythmia.     

肝片吸虫感染对山羊机体自由基代谢影响的研究

实验选择9只健康、6－10月龄雄性白山羊，经粪便学检查和Dot-ELISA检测，确认无肝片吸虫感染，随机分成感染组（n=5)和对照组（n=4)，试验组每只1次口服接种150个肝片吸虫囊蚴，每周定时从颈静脉采集感染前（0周）和感染后15周血液1次分离血清。测定血清谷胱甘肽过氧化物酶（GSH－Px)活性、超氧化物歧化酶（SOD）活性、过氧化物酶（CAT）活性、丙二醛（MDA）和一氧化氮含量的变化，结果表明肝片吸虫感染后血清中GSH－Px活性除第8周试验组显著高于对照组，其他各周无显著差异；在整个实验期间，试验组山羊血清中SOD活性在1－5周呈波动状，以后两组基本处于同一水平；试验组山羊血清CAT活性在感染后第3周显著低于对照组，第6－15周时，试验组始终高于对照组，但各周差异均不显著；试验组山羊血清中MDA含量在整个实验期间始终呈波动状，而在第8、9周试验组显著高于对照组，中周与对照组相比无显著差异；感染后试验组血清中NO开始下降，并且显著低于对照组，至第3－6周时试验组显著高于或低于对照组，第6－15周，试验组一直高于对照组，并在第7、10周时显著地高于对照组。提示山羊感染肝片吸虫后，机体发生多种功能的改变，机体的自由基产生与清除处于相互对抗之中，机体的自由基代谢参与了肝片吸虫病的发展过程。     

Optimizing phosphorus fertilization in an intensive vegetable-rice cropping system

A study was conducted in a sweet pepper-maize-rice cropping system in six farmers fields in Batac, Ilocos Norte, the Philippines, to determine the optimum P fertilizer rate for sweet pepper that will benefit the succeeding crops, maximize system-level productivity and profitability, and reduce the excessive accumulation of P in the soil. Single super phosphate was applied to sweet pepper at rates of 0, 28, 56, 84, 112, and 140 kg P ha^–1 and the succeeding crops were grown without P fertilization. Maize residue was incorporated into the soil at puddling of soil for rice. Phosphorus fertilization at 56 kg P ha^–1 and above had a residual effect on maize and rice. A reduction in the P applied to sweet pepper from 140 to 84 kg P ha^–1 reduced extractable P in the soil at rice harvest from 52 to 29 kg P ha^–1. Phosphorus applied at 111 kg P ha^–1 to sweet pepper was optimum for maximum productivity and economic returns of the sweet pepper-maize-rice cropping system. This rate of P also significantly reduced P accumulation in the soil, thereby reducing the chances of negative effects on soil nutrient balance/availability. The results suggested the need for a cropping systems approach to conserve and effectively use native and fertilizer P in the sweet pepper-rice cropping system.     

Global status of wheat leaf rust caused by <Emphasis Type="Italic">Puccinia triticina</Emphasis>

Leaf rust caused by Puccinia triticina is the most common and widely distributed of the three wheat rusts. Losses from leaf rust are usually less damaging than those from stem rust and stripe rust, but leaf rust causes greater annual losses due to its more frequent and widespread occurrence. Yield losses from leaf rust are mostly due to reductions in kernel weight. Many laboratories worldwide conduct leaf rust surveys and virulence analyses. Most currently important races (pathotypes) have either evolved through mutations in existing populations or migrated from other, often unknown, areas. Several leaf rust resistance genes are cataloged, and high levels of slow rusting adult plant resistance are available in high yielding CIMMYT wheats. This paper summarizes the importance of leaf rust in the main wheat production areas as reflected by yield losses, the complexity of virulence variation in pathogen populations, the role cultivars with race-specific resistance play in pathogen evolution, and the control measures currently practiced in various regions of the world.