Transgene expression in mammary glands of newborn rats

Transgene expression in the mammary glands of newborn rats was studied to establish an early selection system for transgenic animals producing exogenous proteins in their milk during lactation. A fusion gene composed of the bovine alpha S1 casein gene promoter and the human growth hormone gene was microinjected into rat embryos. Transgenic lines that produced human growth hormone in their milk were established and used in this study. Immediately after birth, and without any hormone treatment, human growth hormone was found in the extracts of mammary glands from both male and female rats derived from the line secreting human growth hormone in their milk. The expression of the transgene in mammary glands of newborn rats was also detected by the presence of human growth hormone mRNA. Nontransgenic newborn rats did not express the human growth hormone gene in their mammary glands, while the mRNA for rat alpha casein, an endogenous milk protein, was found in all mammary glands from both transgenic and nontransgenic neonates. These results show that analyzing the expression of transgenes in the mammary glands of neonates is a valuable tool to select the desired transgenic animals and to shorten the selection schedules establishing the transgenic animals. © 1996 Wiley-Liss, Inc.     

Metabolic Reconfiguration in C. elegans Suggests a Pathway for Widespread Sterol Auxotrophy in the Animal Kingdom

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New approaches to estimating the mutagenic potential of chemicals

New developments in mutagenic risk assessment have appeared in the past few years. New methods have been developed such asin vitro micronucleus assay for chromosomal alterations, comet assay for primary DNA damage, use of transgenic animals to detectin vivo gene mutations, and fluorescent in situ hybridization method to detect aneuploidy. Other new methods will be developed in the few next years, including the use of DNA chips and the use of molecular biological methods. Several micromethods have been developed to test a great number of chemical compounds. New concepts have appeared concerning interpretation of data, and particularly of thresholds especially in the case of aneugens; in some cases metabolic or mechanistic thresholds were demonstrated. Genotoxic studies are best integrated into toxicological testing: for example, some genotoxicity tests can be integrated into subacute toxicology; interpretation of data includes metabolism; and toxicokinetic data relate to other toxicological studies. Conversely, genotoxicity data can be used to interpret toxicology studies. This revised version was published online in July 2006 with corrections to the Cover Date.     

Protective immunity induced by vaccination with SAG1 gene-transfected cells against Toxoplasma gondii-infection in mice

To develop a vaccine by augmenting the protective cellular immunity against Toxoplasma gondii (T. gondii)-infection, T gondii SAG1 gene-transfectants were established by using RMA.S (H-2b), a murine transporter associated with the antigen processing (TAP) molecule-deficient lymphoma line, as a host antigen-presenting cell (APC). Immunization of C57BL/6 mice with the SAG1-transfected RMA.S induced CD8+ cytotoxic T lymphocytes (CTL) specific for not only SAG1-transfected RMA.S but also T gondii-infected RMA.S, and elicited protective responses to infection with a virulent T. gondii strain, RH.     

Review of Neospora caninum and neosporosis in animals

Neospora caninum is a coccidian parasite of animals. It is a major pathogen for cattle and dogs and it occasionally causes clinical infections in horses, goats, sheep, and deer. Domestic dogs are the only known definitive hosts for N. caninum. It is one of the most efficiently transmitted parasite of cattle and up to 90% of cattle in some herds are infected. Transplacental transmission is considered the major route of transmission of N. caninum in cattle. Neospora caninum is a major cause of abortion in cattle in many countries. To elicit protective immunity against abortion in cows that already harbor a latent infection is a major problem. This paper reviews information on biology, diagnosis, epidemiology and control of neosporosis in animals.     

Anthrax toxin triggers endocytosis of its receptor via a lipid raft-mediated clathrin-dependent process

The protective antigen (PA) of the anthrax toxin binds to a cell surface receptor and thereby allows lethal factor (LF) to be taken up and exert its toxic effect in the cytoplasm. Here, we report that clustering of the anthrax toxin receptor (ATR) with heptameric PA or with an antibody sandwich causes its association to specialized cholesterol and glycosphingolipid-rich microdomains of the plasma membrane (lipid rafts). We find that although endocytosis of ATR is slow, clustering it into rafts either via PA heptamerization or using an antibody sandwich is necessary and sufficient to trigger efficient internalization and allow delivery of LF to the cytoplasm. Importantly, altering raft integrity using drugs prevented LF delivery and cleavage of cytosolic MAPK kinases, suggesting that lipid rafts could be therapeutic targets for drugs against anthrax. Moreover, we show that internalization of PA is dynamin and Eps15 dependent, indicating that the clathrin-dependent pathway is the major route of anthrax toxin entry into the cell. The present work illustrates that although the physiological role of the ATR is unknown, its trafficking properties, i.e., slow endocytosis as a monomer and rapid clathrin-mediated uptake on clustering, make it an ideal anthrax toxin receptor.     

Production, recovery and immunogenicity of the protective antigen from a recombinant strain of Bacillus anthracis

The protective antigen (PA) is one of the three components of the anthrax toxin. It is a secreted nontoxic protein with a molecular weight of 83 kDa and is the major component of the currently licensed human vaccine for anthrax. Due to limitations found in the existing vaccine formulation, it has been proposed that genetically modified PA may be more effective as a vaccine. The expression and the stability of two recombinant PA (rPA) variants, PA-SNKE-ΔFF-E308D and PA-N657A, were studied. These proteins were expressed in the nonsporogenic avirulent strain BH445. Initial results indicated that PA-SNKE-ΔFF-E308D, which lacks two proteolysis-sensitive sites, is more stable than PA-N657A. Process development was conducted to establish an efficient production and purification process for PA-SNKE-ΔFF-E308D. pH, media composition, growth strategy and protease inhibitors composition were analyzed. The production process chosen was based on batch growth of B. anthracis using tryptone and yeast extract as the only source of carbon, pH control at 7.5, and antifoam 289. Optimal harvest time was 14–18 h after inoculation, and EDTA (5 mM) was added upon harvest for proteolysis control. Recovery of the rPA was performed by expanded-bed adsorption (EBA) on a hydrophobic interaction chromatography (HIC) resin, eliminating the need for centrifugation, microfiltration and diafiltration. The EBA step was followed by ion exchange and gel filtration. rPA yields before and after purification were 130 and 90 mg/l, respectively. The purified rPA, without further treatment, treated with small amounts of formalin or adsorbed on alum, induced, high levels of IgG anti-PA with neutralization activities. Journal of Industrial Microbiology & Biotechnology (2002) 28, 232–238 DOI: 10.1038/sj/jim/7000239 Received 28 August 2001/ Accepted in revised form 20 December 2001     

中国濒危鱼类、两栖爬行类和哺乳类的地理分布格局与优先保护区域——基于《中国濒危动物红皮书》的分析

《中国濒危动物红皮书》依据我国动物所面临的濒危,对我国濒危动物的濒危等级划分,种群现状,致危因素和保护措施等进行了描述说明,首批收录了535种濒危动物。本文以《中国濒危动物红皮书》中所收录的352种濒危脊椎动物（不含鸟类（以下简记为“濒危植物”）为研究对象,整理统计了现有濒危物种的分布资料,在GIS支持下,对中国濒危动物的地理分布进行了研究。结果表明,中国濒危动物物种呈明显的集聚分布,最密集的地区是横断山区,海南岛,西双版纳和云贵高原;而在华北平原,内蒙古东部,黄土高原和东北平原等地区出现大片空白区。影响濒危动物分布的主要因素有热量和水分条件,地形条件等自然条件以及历史开发,人为破坏等人文条件。山地因地形屏障作用而保留了较多的古老物种,其较为复杂的环境因子也有益物种的生存,因而物种丰富度较高。大多数动物对于水分和热量的依赖性较强,因此水热条件对其分布的限制作用十分明显。人为破坏较为严重的地区,濒危物种稀少;污染,开荒等引起的环境问题对于现存动物的威胁很。运用Dobson排除算法得到云南勐腊县等9个县分布有168种濒危动物,占全国总数（海生种类及仅分布于台湾和香港的特有种除外）的51．5％,而其土地面积之和仅为全国陆地总面积的0．9％。而云南勐腊县等94个县市就分布有中国所有的濒危动物。这些地区是我国生物多样性保护应该优先考虑的地方。     

Study on the mechanism of cerium nitrate effects on germination of aged rice seed

Attempts were made to promote germination of naturally and artificially aged rice seeds by treating them with cerium nitrate. The germination rate, germination index, and vigor index of aged rice seed were significantly increased by cerium. It was because the treatments of aged rice seed with cerium nitrate enhanced respiratory rate and activities of superoxide dismutase, catalase, and peroxidase, and decreased superoxide O₂ ⁻ and malondialdehyde contents that plasma membrane permeability was reduced. It was suggested that cerium be used for the seed treatment before sowing.     

羊草草原土壤动物群落多样性的研究

对东北羊草草原不同生境土壤动物群落多样性研究表明 ,生境条件愈优越 ,土壤动物的多样性指数愈高 ,种类则愈丰富 .土壤动物群落的多样性与土壤有机质和全N含量呈正相关 ,与 pH呈负相关 ,与土壤自然含水量和全P含量关系不明显 .土壤动物群落多样性随土层深度的增加而减少 ,且表聚性明显 .