Understanding Animal Companion Surplus in the United States: Relinquishment of Nonadoptables to Animal Shelters for Euthanasia

Relinquishing a nonhuman animal to a shelter is a complex decision that, it is often believed, ultimately may represent a breakdown of the human-animal bond. The result of such a breakdown is an animal companion surplus in the United States, which is no better evidenced than by the statistics documenting the millions of animals euthanized at shelters every year. This research examined the companion animals who are relinquished by their owners to shelters for adoption and compared them specifically to those relinquished for euthanasia. The study also compared the owner characteristics of the animals in these two groups. Although the majority of dogs and cats relinquished are clearly adoptable, a sizable number of these pets presented to shelters for euthanasia have problems precluding their adoptability: old age, illness, and refractory behavior.     

Biochemical Screening of Five Protein Kinases from Plasmodium falciparum against 14,000 Cell-Active Compounds

In 2010 the identities of thousands of anti-Plasmodium compounds were released publicly to facilitate malaria drug development. Understanding these compounds’ mechanisms of action—i.e., the specific molecular targets by which they kill the parasite—would further facilitate the drug development process. Given that kinases are promising anti-malaria targets, we screened ~14,000 cell-active compounds for activity against five different protein kinases. Collections of cell-active compounds from GlaxoSmithKline (the ~13,000-compound Tres Cantos Antimalarial Set, or TCAMS), St. Jude Children’s Research Hospital (260 compounds), and the Medicines for Malaria Venture (the 400-compound Malaria Box) were screened in biochemical assays of Plasmodium falciparum calcium-dependent protein kinases 1 and 4 (CDPK1 and CDPK4), mitogen-associated protein kinase 2 (MAPK2/MAP2), protein kinase 6 (PK6), and protein kinase 7 (PK7). Novel potent inhibitors (IC₅₀ < 1 μM) were discovered for three of the kinases: CDPK1, CDPK4, and PK6. The PK6 inhibitors are the most potent yet discovered for this enzyme and deserve further scrutiny. Additionally, kinome-wide competition assays revealed a compound that inhibits CDPK4 with few effects on ~150 human kinases, and several related compounds that inhibit CDPK1 and CDPK4 yet have limited cytotoxicity to human (HepG2) cells. Our data suggest that inhibiting multiple Plasmodium kinase targets without harming human cells is challenging but feasible.     

基于CRISPR/Cas9基因编辑技术的本科教学实验体系的创建与实践

CRISPR/Cas9是新兴的基因编辑技术,在生命科学研究中发挥着重要的作用。将它引入本科生的实验教学,使本科生了解这项前沿科研技术很有意义。我们创建了一个基于CRISPR/ Cas9技术的本科教学实验体系。该实验体系侧重CRISPR/Cas9技术在哺乳动物细胞中的应用,选用一株基因组上被插入mCherry基因的小鼠胚胎成纤维细胞为实验材料,命名为STO-82。首先设计靶向mCherry的sgRNA,构建CRISPR-Cas9/sgRNA共表达质粒。经测序验证无误后,转染到STO-82细胞。采用流式细胞仪分析检测mCherry阴性和阳性两群细胞,分选出阴性单细胞并扩大培养。最后用测序检验单克隆细胞中靶标DNA序列的编辑情况。结果显示,靶位点有插入或缺失突变,说明体系创建成功。该实验体系将sgRNA设计、CRISPR-Cas9/sgRNA共表达质粒的构建、细胞转染、单细胞分选、单克隆细胞培养、测序序列分析等内容融合为一个综合实验,用于高年级本科生的实验教学。根据实际情况,将教学实践内容分解分块教学,也可以做完整性项目教学。本教学实践采用10人左右的小班分块教学,2人一组,经过3个班（共13组）的实践,绝大部分学生都能完成实验,得到预期结果。通过这个实验,学生加深了对CRISPR/Cas9技术的原理和实验流程的理解,锻炼了实验操作能力和严谨的科研思维,也使学生对该技术的医疗应用风险有了一些认识。     

Changes in serum leptin levels in strenuous exercise and its relation to zinc deficiency in rats

This study aimed to investigate the possible changes in serum leptin concentration caused by acute exercise and the effects of zinc deficiency on these changes. Forty male rats were divided into control-control, control-elercise, zinc-deficient-control, and zinc-deficient-exercise groups (10 rats in each). Control-exercise and zinc-deficient-exercise groups performed exercisse at 6 m/min speed on a rodent treadmill for 60 min or until exhaustion. All rats were decapitated 48h after the exercise, and blood samples were collected to determine serum leptin and zinc levels. Serum leptin levels in the zinc-deficient-control group were lower than in the control-control group. The mean exercise time of control-exercise group was significantly longer than the zinc-deficient-exercise group. We conclude that serum leptin levels significantly decrease both 48 h after strenuous exercise and in the zinc-deficient rats, and there is a further decrease in leptin levels when rats fed on a zinc-deficient diet performed exercise.     

朱砂叶螨对三种杀螨剂的抗性选育与抗性风险评估

为评价朱砂叶螨Tetranychus cinnabarinus对3种杀螨剂的抗性风险,在实验室抗性品系选育基础上,应用数量遗传学中的域性状分析法,研究了朱砂叶螨北碚种群对甲氰菊酯、阿维菌素和哒螨灵3种杀螨剂的抗性现实遗传力,并对3种药剂在不同杀死率下抗性发展的速率进行了预测。结果表明：分别单一连续汰选16代后,朱砂叶螨对甲氰菊酯、阿维菌素的抗性倍数分别达26.54和4.51倍,对哒螨灵表现为敏感性降低（抗性倍数为1.16倍）;朱砂叶螨对甲氰菊酯、阿维菌素和哒螨灵的抗性现实遗传力分别为0.2472,0.1519和0.0160。在室内选择条件下,杀死率为50%～90%时,要获得10倍抗性,甲氰菊酯仅需要13～6代,阿维菌素需要约21～10代;哒螨灵需要约197～89代;在田间选择,三种药剂都将需要更长的时间。抗性筛选16代结果表明,抗性风险较高的是菊酯类的甲氰菊酯,其次是生物源农药阿维菌素,杂环类的哒螨灵抗性风险较小。试验结果可为朱砂叶螨抗性治理提供参考。     

Cotton S-adenosylmethionine decarboxylase-mediated spermine biosynthesis is required for salicylic acid- and leucine-correlated signaling in the defense response to Verticillium dahliae

Planta - Cotton S-adenosylmethionine decarboxylase-, rather than spermine synthase-, mediated spermine biosynthesis is required for salicylic acid- and leucine-correlated signaling in the defense...     

用单性结实基因2A11-iaaM转化罗汉果的研究

为了实现罗汉果生产中免除人工授粉和果实无籽化,该研究利用pBI121-Gus构建果实特异启动子2A11与生长素合成相关基因iaaM的嵌合基因(2A11-iaaM)过量表达载体,以罗汉果雌株叶盘为材料,采用农杆菌介导法建立罗汉果高效遗传转化体系,转化和创制单性结实罗汉果种质,通过基因特异引物对的PCR扩增,初步检测出转基因阳性植株,将之移栽大田,观察转基因植株的单性结实性的表现。结果表明:构建罗汉果单性结实性相关的pBAI-Gus植物双元表达载体获得成功;建立了农杆菌介导的罗汉果叶盘遗传转化优化体系,即农杆菌菌液OD_(600)值为0.3～0.5,侵染10 min,最优选择培养基为MS+TDZ 0.7 mg·L~(-1)+IBA 0.5 mg·L~(-1)+Kan 5 mg·L~(-1)+Cef 300 mg·L~(-1);经PCR鉴定共获得4株转基因阳性雌株;将阳性植株扩繁后移栽田间,经田间调查发现,24株阳性扩繁植株中有5株正常开花,占总植株数的20.8%,且其子房未经人工授粉发育成幼果,表现单性结实性。在载体构建和农杆菌介导的罗汉果遗传转化体系优化的基础上,将外源单性结实相关嵌合基因整合进罗汉果基因组并得到表达,为后续研究单性结实罗汉果的遗传生理,创制转基因罗汉果单性结实新种质,以及克服其产业化中需要人工授粉和无籽化提供了理论和应用基础。     

Myeloperoxidase Polymorphism,Menopausal Status,and Breast Cancer Risk: An Update Meta-Analysis

Myeloperoxidase (MPO) is a metabolic/oxidative lysosomal enzyme secreted by reactive neutrophils at the sites of inflamed organs and tissues during phagocytosis. MPO has been either directly or indirectly linked to neoplasia, which is a well-established risk factor for many types of cancer. A large number of studies have reported the role of MPO G-463A polymorphism regarding breast-cancer risk. However, the published findings are inconsistent. Therefore, we conducted a meta-analysis to determine more precise estimations for the relationship. Eligible studies were identified by searching several electronic databases for relevant reports published before June 2012. According to the inclusion criteria and exclusion criteria, a total of five eligible studies were included in the pooled analyses. When the five eligible studies concerning MPO G-463A polymorphism were pooled into this meta-analysis, there was no evidence found for a significant association between MPO G-463A polymorphism and breast-cancer risk in any genetic model. We also categorized by ethnicity (Caucasian or Asian) for subgroup analysis; according to this subgroup analysis, we found no significant association between MPO G-463A polymorphism and breast-cancer risk in any genetic model. However, in the stratified analysis for the premenopausal group, women carrying the AA genotype were found to have a significantly reduced risk (OR = 0.56, 95% CI 0.34–0.94, p = 0.027). Under the recessive model, there was a significant association between MPO G-463A polymorphism and breast-cancer risk (OR = 0.57, 95% CI 0.34–0.93, p = 0.025). We conclude that MPO-G463A polymorphism might not be a good predictor of breast-cancer risk, though menopausal status modified women’s risk of developing breast cancer.