Sex determination in malaria parasites

A century ago, W. G. MacCallum identified distinct male and female forms in malaria parasites of both birds and humans. Since then, scientists have been puzzled by the high female-to-male ratios of parasites in Plasmodium infections and by the mechanism of sex determination. The sex ratio of malaria parasites was shown to become progressively more male as conditions that allow motility and subsequent fertilization by the male parasites become adverse. This resulted from an increased immune response against male gametes, which coincides with intense host erythropoietic activity. Natural and artificial induction of erythropoiesis in vertebrate hosts provoked a shift toward male parasite production. This change in parasite sex ratio led to reduced reproductive success in the parasite, which suggests that sex determination is adaptive and is regulated by the hematologic state of the host.     

Cross-species interactions between malaria parasites in humans

The dynamics of multiple Plasmodium infections in asymptomatic children living under intense malaria transmission pressure provide evidence for a density-dependent regulation that transcends species as well as genotype. This regulation, in combination with species- and genotype-specific immune responses, results in nonindependent, sequential episodes of infection with each species.     

A cryptic subgroup of Anopheles gambiae is highly susceptible to human malaria parasites

Population subgroups of the African malaria vector Anopheles gambiae have not been comprehensively characterized owing to the lack of unbiased sampling methods. In the arid savanna zone of West Africa, where potential oviposition sites are scarce, widespread collection from larval pools in the peridomestic human habitat yielded a comprehensive genetic survey of local A. gambiae population subgroups, independent of adult resting behavior and ecological preference. A previously unknown subgroup of exophilic A. gambiae is sympatric with the known endophilic A. gambiae in this region. The exophilic subgroup is abundant, lacks differentiation into M and S molecular forms, and is highly susceptible to infection with wild Plasmodium falciparum. These findings might have implications for the epidemiology of malaria transmission and control.     

Genetic loci affecting resistance to human malaria parasites in a West African mosquito vector population

Successful propagation of the malaria parasite Plasmodium falciparum within a susceptible mosquito vector is a prerequisite for the transmission of malaria. A field-based genetic analysis of the major human malaria vector, Anopheles gambiae, has revealed natural factors that reduce the transmission of P. falciparum. Differences in P. falciparum oocyst numbers between mosquito isofemale families fed on the same infected blood indicated a large genetic component affecting resistance to the parasite, and genome-wide scanning in pedigrees of wild mosquitoes detected segregating resistance alleles. The apparently high natural frequency of resistance alleles suggests that malaria parasites (or a similar pathogen) exert a significant selective pressure on vector populations.     

An entomopathogenic fungus for control of adult African malaria mosquitoes

Biological control of malaria mosquitoes in Africa has rarely been used in vector control programs. Recent developments in this field show that certain fungi are virulent to adult Anopheles mosquitoes. Practical delivery of an entomopathogenic fungus that infected and killed adult Anopheles gambiae, Africa's main malaria vector, was achieved in rural African village houses. An entomological inoculation rate model suggests that implementation of this vector control method, even at the observed moderate coverage during a field study in Tanzania, would significantly reduce malaria transmission intensity.     

Chloroquine resistance in Plasmodium falciparum malaria parasites conferred by pfcrt mutations

Plasmodium falciparum chloroquine resistance is a major cause of worldwide increases in malaria mortality and morbidity. Recent laboratory and clinical studies have associated chloroquine resistance with point mutations in the gene pfcrt. However, direct proof of a causal relationship has remained elusive and most models have posited a multigenic basis of resistance. Here, we provide conclusive evidence that mutant haplotypes of the pfcrt gene product of Asian, African, or South American origin confer chloroquine resistance with characteristic verapamil reversibility and reduced chloroquine accumulation. pfcrt mutations increased susceptibility to artemisinin and quinine and minimally affected amodiaquine activity; hence, these antimalarials warrant further investigation as agents to control chloroquine-resistant falciparum malaria.     

Inhibition of development of exoerythrocytic forms of malaria parasites by gamma-interferon

A specific DNA probe was used to study the effect of recombinant rat, mouse, and human gamma-interferon (gamma-IFN) on the course of sporozoite-induced malaria infections. In mice and rats infected with sporozoites of Plasmodium berghei, mouse and rat gamma-IFN's strongly inhibited the development of the exoerythrocytic forms in the liver liver cells of the hosts, but not the development of the erythrocytic stages. The degree of inhibition of the exoerythrocytic forms was proportional to the dose of gamma-IFN administered, but was independent of the number of sporozoites used for challenge. A 30 percent reduction in the development of exoerythrocytic forms in rat liver was achieved when 150 units (about 15 nanograms of protein) of rat gamma-IFN were injected a few hours before sporozoite challenge; the reduction was 90 percent or more with higher doses of gamma-IFN. The effect was less pronounced if the gamma-IFN was administered 18 hours before or a few hours after challenge. Human gamma-IFN also diminished the parasitemia in chimpanzees infected with sporozoites of the human malaria parasite Plasmodium vivax. The target of gamma-IFN activity may be the infected hepatocytes themselves, as shown by in vitro experiments in which small doses of the human lymphokine inhibited the development of exoerythrocytic forms of Plasmodium berghei in a human hepatoma cell line. These results suggest that immunologically induced interferon may be involved in controlling malaria infection under natural conditions.     

Sickle-cell trait in human biological and cultural evolution. Development of agriculture causing increased malaria is bound to gene-pool changes causing malaria reduction

The particular agricultural adaptation we have been considering is the ultimate determinant of the presence of malaria parasites in the intracellular environment of the human red blood cell. This change in the cellular environment is deleterious for normal individuals, but individuals with the sickle-cell gene are capable of changing their red-cell environment so that intense parasitism never develops. Normal individuals suffer higher mortality rates and lower fertility rates in a malarious environment than individuals with the sickle-cell trait do, so the latter contribute proportionately more people to succeeding generations.     

表达人血清白蛋白转基因猪的研究

本研究采用显微注射法，使含有猪血清白蛋白侧翼序列的微小人血清白蛋白导入猪的基因组中，生产出转入血清白蛋白（HSA）基因猪，经PCR和Southern杂交检测，有4头活仔整合有HSA基因，其中的3头不同程度的表达了人血清白蛋白。     

Synthesis of functional human hemoglobin in transgenic mice

Human alpha- and beta-globin genes were separately fused downstream of two erythroid-specific deoxyribonuclease (DNase) I super-hypersensitive sites that are normally located 50 kilobases upstream of the human beta-globin gene. These two constructs were coinjected into fertilized mouse eggs, and expression was analyzed in transgenic animals that developed. Mice that had intact copies of the transgenes expressed high levels of correctly initiated human alpha- and beta-globin messenger RNA specifically in erythroid tissue. An authentic human hemoglobin was formed in adult erythrocytes that when purified had an oxygen equilibrium curve identical to the curve of native human hemoglobin A (Hb A). Thus, functional human hemoglobin can be synthesized in transgenic mice. This provides a foundation for production of mouse models of human hemoglobinopathies such as sickle cell disease.     

转基因棉花连续种植对土壤AM真菌群落结构的影响

为评价转基因棉花种植对土壤中AM真菌群落结构的影响,以转基因棉花013011（抗旱）、SGK321（抗虫）和非转基因棉花TH2（抗旱受体）、石远321（抗虫受体）为材料,研究不同生育期转基因棉花土壤中AM真菌的群落结构。采用PCR-DGGE技术对土壤中AM真菌的群落结构进行分析。结果发现：转基因棉花与非转基因棉花在花铃期和吐絮期土壤中AM真菌群落的最小相似度均大于0.6,这说明了转基因棉花的种植在花铃期和吐絮期均未对土壤中AM真菌的群落结构产生影响。另外转基因棉花和非转基因棉花的种植在花铃期和吐絮期均未对土壤AM真菌的丰富度（S）造成影响;土壤AM真菌香农-维纳指数（H）仅在吐絮期石远321显著低于SGK321,其余品种和时期均未出现显著性差异;土壤AM真菌的均匀度（E_H）仅在吐絮期发现TH2显著高于013011,其余品种和时期均未出现显著性差异。DGGE指纹图谱结果表明转基因棉花与非转基因棉花同一生长时期多为共有条带,且同一时期土壤AM真菌群落结构相似性较高,AM真菌的群落结构无明显变化。两个不同生育期优势属均为Glomus（球囊霉属）。研究表明：土壤AM真菌的群落结构变化只随着棉花生育期的不同会有短暂的变化,与是否为转基因棉花无显著性相关。     

Antigenic diversity in the human malaria parasite Plasmodium falciparum

Monoclonal antibodies against blood forms of Plasmodium falciparum were used to demonstrate considerable antigenic diversity in this species. Different isolates were distinguished by their ability to react with certain antibodies, and most of the antibodies reacted specifically with merozoites, schizonts, or both. The distribution of different antigenic types appeared not to be related to geographic origin. Serological typing with monoclonal antibodies extends the range of methods for identification of different strains of this malaria parasite.     

Allelic exclusion in transgenic mice that express the membrane form of immunoglobulin mu

Antibody-producing cells display a special form of regulation whereby each cell produces immunoglobulin from only one of its two sets of antibody genes. This phenomenon, called allelic exclusion, is thought to be mediated by the product of one heavy chain allele restricting the expression of the other. Heavy chains are synthesized in two molecular forms, secreted and membrane bound. In order to determine whether it is specifically the membrane-bound form of the immunoglobulin M (IgM) heavy chain (mu) that mediates this regulation, transgenic mice were created that carry a human mu chain gene altered so that it can only direct the synthesis of the membrane-bound protein. The membrane-bound form of the human mu chain was made by most of the B cells in these animals as measured by assays of messenger RNA and surface immunoglobulins. Further, the many B cells that express the human gene do not express endogenous mouse IgM, and the few B cells that express endogenous mouse mu do not express the transgene. Thus, the membrane-bound form of the mu chain is sufficient to mediate allelic exclusion. In addition, the molecular structures recognized for this purpose are conserved between human and mouse systems.     

人胰岛素原基因在转基因小鼠乳汁中的表达

将构建的绵羊 β-乳球蛋白基因 ( BLG)调控人胰岛素原基因的重组基因通过显微注射生产转基因小鼠。共移植 888枚注射 BLG-胰岛素原基因的小鼠卵 ,移植受体 31只 ,怀孕 1 4只 ,产仔 53只。PCR检测 51只 ,有 5只为阳性 ,并均为雌性 ,其中 2只小鼠泌乳 ,经放免检测小鼠乳汁中人胰岛素原的质量浓度分别为 37.44mg/L和 39.99mg/L。另外 3只异常消瘦而不孕 ,其中 1只极度衰竭而死亡     

食用菌菌渣的开发利用

主要报道了栽培食用菌后菌渣的营养价值及以其作为食用菌再生产的配料、动物饲料及肥料的研究状况。     

转基因作物产业化发展状况和面临的问题

文章介绍了近年来转基因作物产业化在全球以及国内的发展状况，目前全球种植大豆、玉米、棉花和卡诺拉油菜籽中有五分之一以上是转基因作物，允许种植转基因作物国家的人口已经超过全球人14的50％，我国转基因作物的产业化也获得了较大发展。目前主要存在问题是类型较单一，以及对安全性的争论等。     

Expression of recombinant human butyrylcholinesterase in the milk of transgenic mice

Butyrylcholinesterase (BCHE) is a natural bioscavenger that protects humans against organophosphate toxicity. Due to the limited yield of human BCHE (hBCHE) when purifying from human plasma, it is necessary to find an alternative method to produce this protein. One potential method is to produce transgenic livestock that make modified milk containing high concentration of hBCHE. In this study, we cloned the hBCHEgene into a human lactoferrin (hLF) bacterial artificial chromosome (BAC) construct to make a hLF-hBCHE BAC construct. Subsequently, we injected the BAC construct into pronuclei of mouse fertilized embryos and generated transgenic mice. Expression analysis showed that recombinant hBCHE (rhBCHE) was expressed efficiently in the mammary gland of the transgenic mice and the concentration of rhBCHE in the milk of individual mice ranged from 76±12 to 159±28 mg·L^-1. Protein function tests showed that rhBCHE has the same enzymatic activity as the native hBCHE. Our results pave the way for making transgenic livestock to produce large quantities of rhBCHE.     

Circumsporozoite protein heterogeneity in the human malaria parasite Plasmodium vivax

Phenotypic heterogeneity in the repetitive portion of a human malaria circumsporozoite (CS) protein, a major target of candidate vaccines, has been found. Over 14% of clinical cases of uncomplicated Plasmodium vivax malaria at two sites in western Thailand produced sporozoites immunologically distinct from previously characterized examples of the species. Monoclonal antibodies to the CS protein of other P. vivax isolates and to other species of human and simian malarias did not bind to these nonreactive sporozoites, nor did antibodies from monkeys immunized with a candidate vaccine made from the repeat portion of a New World CS protein. The section of the CS protein gene between the conserved regions I and II of a nonreactive isolate contained a nonapeptide repeat, Ala-Asn-Gly-Ala-Gly-Asn-Gln-Pro-Gly, identical at only three amino acid positions with published nonapeptide sequences. This heterogeneity implies that a P. vivax vaccine based on the CS protein repeat of one isolate will not be universally protective.     

香蕉转基因受体系统的建立

采用横切薄片培养方法建立了香蕉基因转化的适宜受体系统.结果表明:在30℃、黑暗培养15 d,而后转入24℃、光照培养5 d的条件下,香蕉横切薄片的出芽率较高,植株生长较健壮,适宜用作转基因的受体;羧苄青霉素对香蕉薄片的生长没有明显的抑制作用,可以作为转化的抑菌剂;香蕉对潮霉素比较敏感,能够显著抑制香蕉横切薄片的出芽率,确定用40mg.L-1作为潮霉素的筛选含量.