Protein expression signatures: an application of proteomics

The methods of proteomics, the study of the protein complement of the genome, are applicable to environmental testing. Sets of proteins specific to different stressors can be isolated using computer imaging software. Individual proteins can be identified by mass spectrometry. The Protein Expression Signatures (PES) obtained have potential in diagnosing adverse factors in the environment. The challenge is to demonstrate their feasibility in complex environments. We have shown that PES for three endocrine disrupting compounds in trout (Onchorhynchus mykiss), can be detected in mixed sewage effluent. Other studies support these results. As protein databases expand, identification becomes routine, and capture molecules specific to each protein are developed, the possibility of simple field tests for multiple stressors becomes real.     

藻酸双酯钠对蛋白激酶A活性的影响

大鼠肝脏经匀浆、硫酸铵盐析和DEAE-纤维素柱层析,得到部分纯化的Ⅰ型和Ⅱ型蛋白激酶A。将两型酶制剂在体外分别与藻酸双酯钠孵育,藻酸双酯钠对两型酶的活性均有剂量依赖性抑制作用。     

饲料中主要能量物质对大菱鲆幼鱼生长的影响

于1999年6－8月，采用正交设计统计方法，对主要能量物质蛋白质、脂肪和糖类对大菱Ping幼鱼生长的影响进行研究。利用L16（4^3）正交设计了16组大菱Ping幼鱼饲料，在各级饲料中，蛋白质、脂肪和糖类的含量范围分别为36％－45％、8％－20％和0％－6％，共进行了60d的喂养实验，并对饲料及鱼体成分进行分析测定。结果表明，各主要能量物质对鱼体增重和配合饲料转化率的影响顺序为蛋白质＞糖类＞脂肪，且蛋白质的变化对增重率的影响极显著，对饲料转化率的影响显著。     

A comparison of non-hydrolytic methods for extracting amino acids and proteins from coastal marine sediments

Advances in analytical techniques now allow for the potential analysis of intact peptides and proteins isolated from marine sediments. However, there is no established technique for the extraction of macromolecular materials from marine sediments. Six different methods for extracting the amino acid component from coastal marine sediments were compared to the standard hot acid hydrolysis technique for their percent recovery and amino acid composition. The standard hot acid hydrolysis on dried, whole sediments released the greatest concentration of total amino acids (PS-THAA; 3.52 mg gdwt^− 1 ± 10% (SMD)), yet this only accounted for 22% of the total nitrogen in Puget Sound sediments (Washington, USA). Repeated hydrolysis of the same samples did not improve the recovery of nitrogen by more than an additional 10%. Base extraction (0.5 N NaOH) was the second best method for recovering amino acid nitrogen, releasing 60% of the Puget Sound total hydrolyzable amino acids (PS-THAA) (corresponding to 13% of the total sedimentary nitrogen), and has the advantage that it does not rely on peptide hydrolysis to free the nitrogenous component from the sediment matrix. The amino acid distribution of the 0.5 N NaOH extract was not significantly different than the initial THAA. Other non-hydrolyzing methods released lower yields of amino acids (Triton X-100 ≥ hot water > 50 mM NH₄HCO₃ > HF), but might prove to be of use to investigators interested in specific fractions of sedimentary organic nitrogen because these four methods had distinctly different amino acid compositions (enrichments in basic amino acids and depletions in acidic amino acids). Treatments with HF both before and after traditional hydrolysis and/or extractions with base did not release any more of the sedimentary nitrogen. Our results are consistent with the hypothesis that a large fraction of the sedimentary nitrogen (TN) is protected within an organic matrix.     

不同蛋白质水平的饲料对南美白对虾生长的影响

采用鱼粉，豆粉等作为饲料蛋白源，加入等量的混合油（鱼油：豆油＝1：1），制成不同蛋白质梯度的试验饲料，研究不同蛋白质水平饲料对南美白对虾生长的影响，结果表明，不同蛋白质水平饲料显著地影响南美白对虾生长（F＞F0．01＝11．4），饲料中蛋白质为44．12％，能量蛋白比为33．0kJ/g时南美白对虾生长最快，饲料系数最低，体重生长比速最高，通过计算机拟合曲线方程并进行回归分析得出的南美白对虾配合饲料中适宜蛋白质含量为42．37％－44．12％，不同蛋白质水平的饲料对南美白对虾虾体蛋白质含量没有明显的影响。     

Environmental Copper and Manganese in the Pathophysiology of Neurologic Diseases (Alzheimer's Disease and Manganism)

Alzheimer's disease (AD) is the most common form of dementia. Populations migrating from developing to industrialized countries seem to elicit a higher incidence and prevalence rate of AD, suggesting lifestyle and environmental factors to have a role in the pathophysiology of AD. One of its major neuropathological hallmarks is the deposition of Aβ peptides as amyloid plaques in the brain of AD patients. Aβ is proteolytically cleaved out of the larger amyloid precursor protein (APP). Cu and Mn are often found in drinking water and may have a neurotoxic potential. APP is involved in Cu homeostasis in mouse and man. In vitro observations and in vivo data obtained from APP mouse models provide strong evidence that APP overexpression enables intracellular Cu to be transported out of the cell. Disturbed metal‐ion homeostasis with elevated serum Cu levels occurs in Alzheimer and Down's patients and lowered levels in post‐mortem AD brain. We observed that bioavailable Cu has specific beneficial effects in an Alzheimer's disease mouse model. This should be regarded as a proof‐of‐concept for a prophylactic approach to overcome the observed CNS Cu deficiency in the brain of Alzheimer's disease patients. Manganism is a disorder with symptoms similar to that of Parkinson's disease. The precise mechanism how manganese can damage the nervous system is unknown. There is some evidence that iron and manganese may utilize similar transport systems. Epidemi ologic data strongly suggests that manganese enters the body primarily via inhalation and through the ingestion of manganese in drinking water.     

重组质粒HF443-EGFP的构建及其在赤潮毒素检测中的应用初探

采用基因工程的方法，进行重组质粒HF443一EGFP的构建和在哺乳动物细胞中的表达，并进行了其用于赤潮毒素检测的可能性研究。结果表明，用PCR技术扩增EGFP基因，用XbaⅠ、Hind Ⅲ双酶切并纯化，与同样双酶切并纯化的HF443质粒连接，可构建重组质粒HF443-EGFP；利用Lipofect AMINE2000转染体外培养哺乳动物BIU-87细胞。在培养体系中加入不同浓度的GTX，而后加入Na^ 通道激活剂乌头碱以诱导绿色荧光蛋白表达，荧光显微镜下观察细胞中绿色荧光蛋白表达的变化情况。细胞发出的荧光随毒素浓度的增加而减弱，存在一定的剂量效应关系，提示毒素GTX可显著影响绿色荧光蛋白的表达，重组质粒：HF443-EGFP用于赤潮霉素检测有一定的可行性.     

黄河三角洲河蚬(Corbicula fluminea)的繁殖生物学研究

自2009年2月—2010年1月,共采集黄河三角洲流域分布的河蚬498个,采用组织学方法分析其性腺的年周期变化,利用考马斯亮蓝方法进行了蛋白质含量的年周期测量。结果表明,黄河三角洲分布的河蚬雌雄异体,性比为1:1,性腺发育分为6个时期:增殖期、生长期、成熟期、排放期、恢复期和排尽休止期。配子发生期开始于2月份,一年内有两个繁殖高峰期,分别为每年的6月中旬和10月上旬。蛋白质含量无性别差异,闭壳肌和外套膜的蛋白质含量全年都处于较低水平,无明显波动,性腺-内脏团和足的蛋白质含量相对较高,并分别在5月份和5、8月份达到峰值,与河蚬的繁殖盛期显著相关。     

CORNELL DESIGN APPLIED ON OPTIMAL MACRONUTRIENT COMPOSITION OF FORMULATED DIETS FOR DAVID＇S SCHIZOTHORACIN [SCHIZOTHORAX （RACOMA） DAVID1] JUVENILES

采用Comell方法研究了初始重为（2．0±0．1）g的重口裂腹鱼幼鱼日粮中常量营养物质的适宜需求量。日粮中蛋白质、脂肪和碳水化合物梯度变化范围分别为33％-51％、3％-18％和24％-33％,试验周期为60d。试验结果表明,除日粮蛋白质33％组鱼成活率明显下降外（P〈0．05）,其它试验日粮中3种常量营养物质含量对重口裂腹鱼幼鱼成活率没有显著影响（P〉0．05）。当日粮蛋白质水平≤45％时,其特定生长率随着日粮蛋白质水平上升而明显增加;当日粮蛋白质水平在33％-36％时,重口裂腹鱼的特定生长率显著降低（P〈0．05）。除12％日粮脂肪组试验鱼外,9％日粮脂肪组试验鱼的特定生长率显著性高于其它脂肪水平日粮组（P〈0．05）,但当日粮中脂肪含量≥15％后,重口裂腹鱼鱼体脂肪含量显著上升（P〈0．05）,肝细胞脂肪变性明显。日粮中不同的碳水化合物水平对重口裂腹鱼特定生长率没有显著影响（P〉0．05）,但当日粮中碳水化合物为33％时,鱼体糖原含量显著增加（P〈0．05）。试验表明,重口裂腹鱼幼鱼日粮蛋白质、脂肪和碳水化合物3种常量营养物质的适宜需求量分别为45％、9％和≤30％。日粮总能为19．0MJ／kg,蛋白质能量比（P／E）为23mg／J。     

Growing Spirodela polyrrhiza in Swine Wastewater for the Production of Animal Feed and Fuel Ethanol: A Pilot Study

To evaluate the performance of Spirodela polyrrhiza grown in swine wastewater for protein and starch production under field conditions, a pilot‐scale duckweed culture pond was installed at Barham Farm, Zebulon, North Carolina and operated from May to November 2010. The anaerobically treated swine wastewater was fed to the duckweed pond intermittently to provide nutrients for the growth of duckweed, and the duckweed biomass was harvested regularly from the pond and prepared as a protein‐ or starch‐rich feedstock for the production of animal feed or fuel ethanol. Over the experimental period, the duckweed pond produced protein and starch at rates of 2.68 and 1.88 g m^?2 day^?1, respectively. During the same time, NH₄–N and o‐PO₄–P in the wastewater were, respectively, removed at rates of 92.9 and 2.90 mmol m^?2 day^?1.