Effect of 4-chloro-2-methylphenoxyacetic acid and 2,4-dimethylphenol on human erythrocytes

The effects of exposure of human erythrocytes to different concentrations of 4-chloro-2-methylphenoxyacetic acid (MCPA) and its metabolite—2,4-dimethylphenol (2,4-DMP) were studied. The investigations concerned mainly the content of glutathione (GSH and GSSG), glutathione peroxidase (GSH-Px), glutathione transferase (GST), and the level of adenine energy charge (AEC). Reactive oxygen species (ROS) such as hydroxyl radical, superoxide anion, hydrogen peroxide, and nitric oxide are produced during normal processes in the cell. Under normal conditions, antioxidant systems of the cell minimize damage caused by ROS. When ROS generation increases to an extent that it overcomes the cellular antioxidant systems, the result is oxidative stress. We observed that MCPA and 2,4-DMP decreased the level of GSH in erythrocytes in comparison with control. MCPA did not affect glutathione peroxidase and glutathione transferase activity, while 2,4-DMP increased their activity. 2,4-DMP decreased the level of ATP and increased the content of ADP and AMP, leading to the fall of the level of AEC. MCPA and 2,4-DMP transform hemoglobin into methemoglobin, thus preventing oxygen transport. Comparison of the toxicity of MCPA and 2,4-DMP revealed that the most prominent changes occurred in human erythrocytes incubated with 2,4-DMP.     

肉苁蓉对感染性休克肝线粒体功能影响的实验研究

目的:研究肉苁蓉对感染性休克肝线粒体功能的影响,探讨肉苁蓉在细胞水平治疗感染性休克的机制.方法:40只SD大鼠随机分为5组:假手术组,12 h对照组,12 h给药组,16 h对照组和16 h给药组.采用盲肠结扎穿孔术(CLP)复制感染性休克模型,比较各组存活率和平均动脉压(MAP),肝线粒体ATP酶活性,呼吸控制率(RCR),ADP/O比值的变化.结果:大鼠在复制感染性休克模型后,肝线粒体ATP酶活性,呼吸控制率和ADP/O比值显著降低,24 h存活率明显下降.给药组肝线粒体ATP酶活性,呼吸控制率(RCR)和ADP/O比值较对照组明显改善(P＜0.05),与大鼠死亡率的降低呈明显正相关.结论:肉苁蓉能增强线粒体摄氧功能和氧化磷酸化功能,恢复感染性休克线粒体的正常的能量代谢功能.     

Platelet function in dogs: breed differences and effect of acetylsalicylic acid administration

BACKGROUND: Clinical studies investigating platelet function in dogs have had conflicting results that may be caused by normal physiologic variation in platelet response to agonists. OBJECTIVES: The objective of this study was to investigate platelet function in clinically healthy dogs of 4 different breeds by whole-blood aggregometry and with a point-of-care platelet function analyzer (PFA-100), and to evaluate the effect of acetylsalicylic acid (ASA) administration on the results from both methods. METHODS: Forty-five clinically healthy dogs (12 Cavalier King Charles Spaniels [CKCS], 12 Cairn Terriers, 10 Boxers, and 11 Labrador Retrievers) were included in the study. Platelet function was assessed by whole-blood aggregation with ADP (1, 5, 10, and 20 microM) as agonist and by PFA-100 using collagen and epinephrine (Col + Epi) and Col + ADP as agonists. Plasma thromboxane B(2) concentration was determined by an enzyme immunoassay. To investigate the effect of ASA, 10 dogs were dosed daily (75 or 250 mg ASA orally) for 4 consecutive days. RESULTS: A higher platelet aggregation response was found in CKCS compared to the other breeds. Longer PFA-100 closure time (Col + Epi) was found in Cairn Terriers compared to Boxers. Plasma thromboxane B(2) concentration was not statistically different between groups. Administration of ASA prolonged the PFA-100 closure times, using Col + Epi (but not Col + ADP) as agonists. Furthermore, ASA resulted in a decrease in whole-blood platelet aggregation. CONCLUSIONS: Platelet function is influenced by breed, depending upon the methodology applied. However, the importance of these breed differences remains to be investigated. The PFA-100 method with Col + Epi as agonists, and ADP-induced platelet aggregation appear to be sensitive to ASA in dogs.     

小麦白粉菌ADP/ATP载体蛋白基因的克隆及表达特征分析

 小麦白粉病是小麦上的一种重要病害。研究小麦白粉菌致病相关基因及其在致病过程中的作用,对于丰富小麦白粉菌致病的分子机制和筛选防治新靶标具有重要意义。前期转录组测序结果提示一个小麦白粉菌ADP/ATP载体蛋白（ADP/ATP carrier protein,AACP）在小麦与白粉菌互作时上调表达。本研究利用cDNA末端快速克隆技术（rapid\|amplification of cDNA ends,RACE）获得了长945 bp具有完整开放阅读框的小麦白粉菌ADP/ATP carrier基因序列,暂命名为BgtAACPx（GenBank登录号为MF197857）该基因编码314个氨基酸残基,利用相关软件进行生物信息分析,结果表明该蛋白为疏水性,含有6个跨膜区,亚细胞定位在线粒体上。与已有小麦白粉菌AACP蛋白的同源性为97%,是一个新的蛋白,同时构建了与其同源蛋白的遗传进化树。定量结果表明该基因在小麦白粉菌吸器期（48 h）、次生菌丝（72 h）至白粉菌刚产孢子又未形成孢子堆（120 h）这段时期高表达,提示该基因可能参与小麦白粉菌对小麦的致病过程。     

Platelet aggregation responses in clinically healthy adult llamas

Background: Limited information exists regarding hemostasis in camelids despite the importance of platelet function testing in the accurate identification of platelet disorders. As further importation of llamas to North America is restricted, variability in breeding stock will continue to decrease, potentially leading to an increase in heritable bleeding disorders. Objective: The objective of this study was to measure platelet aggregation responses in clinically healthy llamas and provide baseline data to which abnormal platelet function may be compared in the future. Methods: Blood samples were collected from 39 healthy adult llamas, citrated, and centrifuged to produce platelet‐rich plasma (PRP). Within 4 hours of the blood draw, 20 μL of each agonist reagent were added to 180 μL of PRP. Final concentrations of agonists were 2 × 10^?5 M ADP, 0.19 mg collagen/mL PRP, 1 × 10^?4 M epinephrine, and 500 μg arachidonic acid/mL PRP. Results: Llama platelets were most responsive to ADP and collagen, with a maximum percent aggregation (mean±SD) of 71.3±18.6% and 55.8±19% and aggregation rates of 9.5±3.9 and 6.7±3.7 cm/min, respectively. Llama platelet aggregation in response to epinephrine and arachidonic acid was minimal to absent. Conclusions: This study is the first of its kind to establish baseline values for platelet aggregation in healthy adult llamas.     

小麦TaAAC基因克隆及功能的初步验证

为明确ATP/ADP转运蛋白在小麦籽粒粒重及淀粉合成方面的作用,以百农207 cDNA为模板,利用RT-PCR技术获得TaAAC的基因全长。生物信息学分析表明,TaAAC基因CDS长度为1 143 bp,编码380个氨基酸;TaAAC蛋白位于线粒体内膜和质体膜上。随后,以大麦条纹花叶病毒(BSMV)为载体,以GFP基因为指示基因,利用VIGS和qRT-PCR技术对TaAAC基因的功能进行研究。结果表明,在病毒诱导接种后24 d和29 d,TaAAC基因的相对表达量分别较对照下降51%和67%。BSMV病毒诱导TaAAC基因沉默后的百农207籽粒内总淀粉含量和千粒重均较对照显著降低。以上结果表明TaAAC基因正向调控小麦籽粒的淀粉合成,提高该基因表达有助于增加小麦粒重。     

Investigating the in situ degradation of atrazine in groundwater

This study focused on whether or not atrazine could be degraded by indigenous groundwater bacteria as part of an in situ remediation approach. Groundwater was taken from an unconfined middle upper chalk site where concentrations of atrazine and nitrate were typically in the ranges 0.02-0.2 microg litre-1 and 11.6-25.1 mg NO3-N litre-1 respectively. Sacrificial batch studies were performed using this groundwater spiked with atrazine at a concentration of 10 microg litre-1 in conjunction with a minimal mineral salts liquid (Glu-MMSL) medium which contained glucose as the sole carbon source. Treatments comprised either the Glu-MMSL groundwater cultured bacteria or Pseudomonas sp. strain ADP. Results from sacrificial batches indicated the occurrence of bacterial growth and denitrification as monitored by optical density (absorbance at 600 nm) and NO3-N content. Analysis of atrazine content by solid phase extraction coupled with high-performance liquid chromatography showed no degradation of atrazine over a period of 103 days in either treatment. These results indicated that no acclimatised bacterial community featuring positive degraders to the herbicide atrazine had become established within this chalk aquifer in response to the trace levels encountered.     

小麦叶片线粒体的分离

用差速离心法分离绿色小麦叶片线粒体。结果表明,经两次低速离心和两次高速离心,分离所得线粒体具有较高的呼吸活性和ADP/O比值,线粒体完整率达70％左右。本文对影响线粒体分离效果的因素亦进行了讨论。     

野菊花抗血小板聚集有效成分的筛选

以野菊花为材料,顺次用石油醚、氯仿、乙酸乙酯、正丁醇、水抽提,抽提物终浓度相当于1g草药/ml,对5μmol/L ADP诱导的血小板聚集分别抑制30.8±20.2％、49.3±29.4％、51.9±22.8％、43.5±18.0％、13.7±1.6％。野菊花乙酸乙酯抽提物抑制5μmol/L ADP诱导的血小板聚集IC_(50)=1mg/ml。乙酸乙酯抽提物用硅胶柱制备分离得C1、C2、C3、C4、C5、C6六种结晶,经TLC检识分别为一个斑点。定性分析知为基黄酮化合物。在0.5mg/ml浓度下,对2μmol/L ADP诱导的血小板聚集后三种物质分别抑制78.9％、85.3％、83.5％。     

Measurement of platelet aggregation in ovine blood using a new impedance aggregometer

Background: Whole blood platelet aggregometry (impedance) is an important method to investigate platelet function disorders. Examination of hemostatic function in sheep is important with respect to their role as an animal model of human disease. Objective: The aim of this study was to evaluate and optimize selected methodological aspects (anticoagulant, agonist concentration) of impedance aggregometry in ovine blood using the new Multiplate 5.0 analyzer. Methods: Blood samples were collected in hirudin anticoagulant from 40 clinically healthy sheep. Samples from selected sheep were collected in citrate, with or without the addition of calcium chloride. The agonists adenosine diphosphate (ADP), collagen, ristocetin, arachidonic acid, and thrombin receptor‐activating peptide (TRAP) were added in several concentrations to induce aggregation. Results: Based on maximum aggregation values and internal precision, no significant difference was found between ADP concentrations of 3–10 μmol/L and collagen concentrations of 3–5 μg/mL (P>.05). The lowest interindividual variation of approximately 3–4‐fold was seen with 4 and 5 μmol/L ADP and 4 and 5 μg/mL collagen. Ristocetin, arachidonic acid, and TRAP did not induce significant aggregation at any concentration. Aggregation results were significantly lower when measured in citrate‐ vs hirudin‐anticoagulated blood, regardless of the presence of calcium chloride. Conclusions: Our results indicate that the multiplate impedance aggregometer is suitable for the measurement of platelet aggregation in sheep using optimal agonist concentrations of 4–5 μmol/L ADP and 4–5 μg/mL collagen. Hirudin‐anticoagulated blood is the preferred sample material.