Visualization of the mitochondria ofToxoplasma gondii-infected mouse fibroblasts by the cationic permeant fluorescent dye rhodamine 123

Summary The mitochondria of living mouse fibroblasts infected withToxoplasma gondii were monitored with the cationic permeant fluorescent dye rhodamine 123. Fluorescence microscopy revealed that host cell mitochondria accumulated at the cytoplasmic surface of parasitophorous vacuoles and increased the dye uptake in the periparasitophorous vacuole asT. gondii multiplied.The author wishes gratefully to acknowledge the work of Miss Mayumi Kato and Mr. Isao Kimata in preparing the photographs. The publication cost was supported by a grant of the Ohyama Health Foundation.     

The course ofPlasmodium berghei infection in mice latently infected withToxoplasma gondii

Summary The course of infection with 2 different virulent strains ofPlasmodium berghei was investigated in mice latently infected withToxoplasma gondii. When given the highly virulent ANKA strain ofP. berghei all Toxoplasma-infected mice died but the survival time was prolonged. After infection with the less virulent strain K 173 mice could survive the subsequent infection. In these cases levels of parasitemia depended upon the duration of theT. gondii infection. Mice infected for about 6 weeks withT. gondii showed maximum protection. These studies were conducted in the Institut für Medizinische Parasitologie der Universit?t Bonn (D-5300 Bonn, Federal Republic of Germany).     

Antibody-induced formation of caps inToxoplasma gondii

Summary Trophozoites ofToxoplasma gondii from mouse peritoneal exudate move their surface membrane antigens towards one pole of the cell when incubated with antibodies. The phenomenon may be induced in up to 50% of incubated parasites. It is prvented by some metabolic inhibitors and low temperatures (0–4°C). These properties do not change in parasites subpassaged after repeated incubation with antibodies.We wish to thank Dr. J.lusarczyk of the National Institute of Hygiene for his help in preparation the photomicropgraphs.     

Host cell manipulation by the human pathogen Toxoplasma gondii

Toxoplasma gondii is an obligate intracellular parasite that can infect virtually any nucleated cell. During invasion Toxoplasma creates the parasitophorous vacuole, a subcellular compartment that acts as an interface between the parasite and host, and serves as a platform for modulation of host cell functions that support parasite replication and infection. Spatial reorganization of host organelles and cytoskeleton around the parasitophorous vacuole are observed following entry, and recent evidence suggests this interior redecorating promotes parasite nutrient acquisition. New findings also reveal that Toxoplasma manipulates host signaling pathways by deploying parasite kinases and a phosphatase, including at least two that infiltrate the host nucleus. Toxoplasma infection additionally controls several cellular pathways to establish an anti-apoptotic environment, and subverts immune cells as a conduit for dissemination. In this review we discuss these recent developments in understanding how Toxoplasma achieves widespread success as a human and animal parasite by manipulating its host.     

刚地弓形虫RH株感染对BALB／c小鼠情绪行为的影响

目的研究刚地弓形虫RH株感染对BALB／c小鼠学习记忆行为的影响及可能机制。方法将72只周龄、大小相近的雄性BALB／c小鼠采用随机数字表分为生理盐水对照组与不同数量（3×10^3／ml、3×10^4／ml、3×10^5／m1）弓形虫RH株感染组,每组18只。于感染第5周从每组各随机抽取6只分别进行高架十字迷宫实验、旷场实验及强迫游泳实验,观察各组小鼠在实验中情绪行为变化。并记录各项指标进行统计分析。结果在高架迷宫试验与旷场实验中,3×10^3／ml弓形虫感染组小鼠与生理盐水对照组小鼠比较,各项行为学指标无明显差异。在3×10^4／ml、3×10^5／ml弓形虫感染组小鼠与对照组小鼠比较出现明显降低（P〈0．05）．以3×10^5／ml感染组最为明显,其小鼠运动活力（0E＋cE）、进入开放臂次数比例（OE％）、开放臂停留时间比例（OT％）小鼠爬行总格数、中央格在总格数中比例分别为11．08±2．12、28．73±0．59％、25．62±2．33％、32．30±17．26、2．42±0．65％。在强迫游泳试验中,各弓形虫感染组小鼠游泳的静止时间均高于对照组小鼠（P〈0．05）,3×10^5／ml感染组静止时间最长,达226．6±1．9S。结论刚地弓形虫RH株感染可引起小鼠情绪行为改变,具有焦虑抑郁倾向。     

刚地弓形虫培养上清抑制THP-1细胞增殖及诱导凋亡的研究

摘要：目的提取弓形虫体外细胞共培养上清,并研究上清对人急性单核细胞白血病细胞THP-1增殖及凋亡的影响。方法收集对数生长期的THP-1细胞以5X10^7／ml细胞浓度接种于不同培养瓶中,对照组加入含10％胎牛血清的RPMll640,实验组加入相同体积不同数量（2×10^7／ml、4X10^7／ml、8×10^7／m1）弓形虫速殖子培养上清,采用四甲基氮噻唑蓝（MTY）法检测吸光度（A490值）并计算THP-1细胞增殖抑制率;倒置显微镜下观察细胞形态变化;Annexin-V-FITC／PI染色细胞后上流式细胞仪检测各个时间点细胞凋亡率变化,以Western印迹方法分析凋亡相关蛋白Bax、Bcl-2的表达或活性。结果MTY法检测结果弓形虫培养上清呈时间剂量依赖性抑制THP-1细胞株增殖,倒置显微镜下观察处理组细胞有发泡现象和凋亡小体出现。流式细胞仪检测弓形虫感染后的THP-1细胞凋亡率较对照组有升高趋势（P〈0．05）,呈量效依赖性,Westernblot检测刚地弓形虫培养上清作用于THP-l细胞48h后实验组的Bax、Bcl-2蛋白表达较对照组的比值分别有明显的升高与降低（P〈0．05）。结论刚地弓形虫速殖子培养上清对体外培养THP-l细胞增殖有明显的抑制作用,并可诱导THP-1细胞凋亡。     

Suppression and enhancement of humoral immune response to Toxoplasma gondii by passive antibody

Anti-toxoplasma antibodies administered passively to mice may lead to suppression of enhancement (reported for the first time with Protozoan parasites) of subsequent antibody response when these animals are later infected with Toxoplasma gondii. The outcome is dependent on infecting strain of Toxoplasma and the antigen-antibody ratio.     

Visualization of the mitochondria of Toxoplasma gondii-infected mouse fibroblasts by the cationic permeant fluorescent dye rhodamine 123

The mitochondria of living mouse fibroblasts infected with Toxoplasma gondii were monitored with the cationic permeant fluorescent dye rhodamine 123. Fluorescence microscopy revealed that host cell mitochondria accumulated at the cytoplasmic surface of parasitophorous vacuoles and increased the dye uptake in the periparasitophorous vacuole as T. gondii multiplied.     

A latent ability to persist: differentiation in <Emphasis Type="Italic">Toxoplasma gondii</Emphasis>

A critical factor in the transmission and pathogenesis of Toxoplasma gondii is the ability to convert from an acute disease-causing, proliferative stage (tachyzoite), to a chronic, dormant stage (bradyzoite). The conversion of the tachyzoite-containing parasitophorous vacuole membrane into the less permeable bradyzoite cyst wall allows the parasite to persist for years within the host to maximize transmissibility to both primary (felids) and secondary (virtually all other warm-blooded vertebrates) hosts. This review presents our current understanding of the latent stage, including the factors that are important in bradyzoite induction and maintenance. Also discussed are the recent studies that have begun to unravel the mechanisms behind stage switching.     

Chemical and tactile communication between the root aphid parasitoidParalipsis enervis and trophobiotic ants: consequences for parasitoid survival

Females of the aphid parasitoidParalipsis enervis received liquid food by regurgitation (trophallaxis) from workers of the ant speciesLasius niger, but were not fed by workers ofMyrmica laevinodis andTetramorium caespitum. WhileP. enervis was not treated aggressively by workers of any of these species,Lasius flavus workers killed the parasitoid. This different ant behaviour resulted in a different parasitoid longevity. WhileP. enervis survived for only 10 min in the presence ofL. flavus (due to ant aggression) or for approximately one day in the presence ofT. caespitum andM. laevinodis (due to lack of trophallaxis), survival increased significantly to more than five days in the presence ofL. niger, which provided food regularly to the parasitoids. Our study suggests thatP. enervis mimics behavioural signals ofL. niger, as well as odor cues of its host aphidAnoecia corni, to avoid aggression byL. niger.     

<Emphasis Type="Italic">Toxoplasma gondii</Emphasis> chromosomal passenger complex is essential for the organization of a functional mitotic spindle: a prerequisite for productive endodyogeny

The phylum Apicomplexa encompasses deadly pathogens such as malaria and Cryptosporidium. Apicomplexa cell division is mechanistically divergent from that of their mammalian host, potentially representing an attractive source of drug targets. Depending on the species, apicomplexan parasites can modulate the output of cell division, producing two to thousands of daughter cells at once. The inherent flexibility of their cell division mechanisms allows these parasites to adapt to different niches, facilitating their dissemination. Toxoplasma gondii tachyzoites divide using a unique form of cell division called endodyogeny. This process involves a single round of DNA replication, closed nuclear mitosis, and assembly of two daughter cells within a mother. In higher Eukaryotes, the four-subunit chromosomal passenger complex (CPC) (Aurora kinase B (ARKB)/INCENP/Borealin/Survivin) promotes chromosome bi-orientation by detaching incorrect kinetochore–microtubule attachments, playing an essential role in controlling cell division fidelity. Herein, we report the characterization of the Toxoplasma CPC (Aurora kinase 1 (Ark1)/INCENP1/INCENP2). We show that the CPC exhibits dynamic localization in a cell cycle-dependent manner. TgArk1 interacts with both TgINCENPs, with TgINCENP2 being essential for its translocation to the nucleus. While TgINCENP1 appears to be dispensable, interfering with TgArk1 or TgINCENP2 results in pronounced division and growth defects. Significant anti-cancer drug development efforts have focused on targeting human ARKB. Parasite treatment with low doses of hesperadin, a known inhibitor of human ARKB at higher concentrations, phenocopies the TgArk1 and TgINCENP2 mutants. Overall, our study provides new insights into the mechanisms underpinning cell cycle control in Apicomplexa, and highlights TgArk1 as potential drug target.     

Dominance for enzyme activity inDrosophila melanogaster

Summary A regulatory element tightly linked to theGpdh locus inDrosophila melanogaster has been isolated from a natural population. Flies homozygous for second chromosomes bearing the element,H31, have half the GPDH activity of normal homozygotes. Heterozygotes betweenH31 andF orS alleles exhibit dominance in GPDH activity. Heterozygotes betweenH31, F orS andDf(2L) GdhA have half the diploid level. The contribution of theS allele to the activity inS/H31 heterozygotes is more than four times that ofH31. The regulatory element distinguishingH31 is tightly linked to theGpdh ⁺ locus.     

Monogenic determinants of familial Alzheimer's disease: presenilin-2 mutations

Presenilin-2 (PS2) is one of three genes [amyloid precursor protein (APP), presenilin-1 (PS1) and PS2] shown to cause familial Alzheimer's disease (FAD), and is highly homologous to PS1. Currently demonstrated functions of PS2 include interactions with APP and Aβ, and participation in apoptotic pathways. PS2 FAD mutations influence APP processing in a manner predicted to promote amyloid formation and also enhance the proapoptotic effect of wild-type PS2. Other possible functions of PS2 are related to its homology to Notch pathway genes in Caenorhabditis elegans, suggesting it may have a developmental role. PS2-associated AD is the most reminiscent of the sporadic form of the disease in terms of older age of onset and longer disease duration. Since PS2 mutations are incompletely penetrant and age of onset in carriers is highly variable (40 – 88 years), elucidation of PS2 mechanisms may reveal factors which modify AD and are therapeutically relevant to sporadic AD.     

Forecasting for the generation of trading signals in financial markets

In this paper we show that optimal trading results can be achieved if we can forecast a key summary statistic of future prices. Consider the following optimization problem. Let the return r_i (over time i=1, 2, ..., n) for the ith day be given and the investor has to make investment decision d_i on the ith day with d_i=1 representing a ‘long' position and d_i=0 a ‘neutral' position. The investment return is given by r=Σⁿ_i=1r_id_i−cΣⁿ⁺¹_i=1∣d_i−d_i−1∣, where c is the transaction cost. The mathematical programming problem of choosing d₁, ..., d_n to maximize r under a given transaction cost c is shown to have an analytic solution, which is a function of a key summary statistic called the largest change before reversal. The largest change before reversal is recommended to be used as an output in a neural network for the generation of trading signals. When neural network forecasting is applied to a dataset of Hang Seng Index Futures Contract traded in Hong Kong, it is shown that forecasting the largest change before reversal outperforms the k‐step‐ahead forecast in achieving higher trading profits. Copyright © 2000 John Wiley & Sons, Ltd.     

Short-range linkage relationships, genomic organisation and sequence comparisons of a cluster of five HSP70 genes in Fugu rubripes

Twelve cosmids containing sequences resembling genes encoding members of the 70-kDa heat-shock protein family, HSP70, have been isolated from Fugu rubripes. They can be broadly divided into three groups of overlapping cosmids. Restriction analysis and sequencing of one set of five cosmids have revealed five intronless Fugu HSP70 genes spanning 42 kb, arranged in a combined head-to-head, tail-to-tail and head-to-tail orientation. The levels of DNA and amino acid identity are very high with respect to one another, and are most similar to HSP70 sequences linked to the major histocompatibility complex (MHC) region in other species. Putative heat-shock consensus elements are identified. Non-HSP70 sequences with homology to known genes have been found physically linked to this Fugu HSP70 cluster: the Drosophila melanogaster SOL gene, the Drosophila melanogaster nemo gene, the Caenorhabditis elegans T17E9.1 gene and the sequence encoding the serine protease domain. The linkage relationships described here so far bear no resemblance to those of HSP70 in other organisms. Convergence of mammalian HSP70 and MHC class I and II loci probably occurred after fish had diverged. Received 17 November 1998; received after revision 25 February 1999; accepted 26 February 1999     

Notable diversity in hemoglobin expression patterns among species of the deep-sea clam, <Emphasis Type="Italic">Calyptogena</Emphasis>

The deep-sea clams Calyptogena nautilei and C. tsubasa, which live in the cold-seep area at a depth of 3570 m in the Nankai Trough, Japan, have abundant hemoglobins (Hbs) in erythrocytes, similar to other Calyptogena species. We determined the cDNA-derived amino acid sequences of Hbs from two Calyptogena species. C. tsubasa was found to contain two dimeric Hbs, Hb I consisting of 145 amino acid residues and Hb II with 137 residues, similar to known Hbs from C. soyoae and C. kaikoi. Sequence identity was over 90% among the orthologous chains of Calyptogena Hbs. On the other hand, surprisingly, C. nautilei contained two monomeric Hbs, Hb III containing 141 residues and Hb IV with 134 residues. In addition, Hbs III and IV showed only 33–42% sequence identity with Hbs I and II from other Calyptogena species. The distal (E7) histidine, one of the functionally important residues of the heme protein, is replaced by glutamine in all Hb chains of Calyptogena species. A phylogenetic analysis indicated that C. nautilei Hb III is closer to Hb I from other Calyptogena species. We suppose that a Hb gene was duplicated at least three times in an immediate ancestor of Calyptogena and, presumably depending on physiological conditions different Hb sets are being expressed: dimeric Hbs I and II in C. soyoae, C. kaikoi and C. tsubasa, and monomeric Hbs III and IV in C. nautilei. Received 13 May 2003; received after revision 5 June 2003; accepted 12 June 2003     

Caryotypes de quelques hybrides interspécifiques dePitymys (Mammalia,Rodentia)

Summary The Robertsonian polymorphism ofPitymys subterraneus involves only 1 pair of metacentric autosomes. The crosses of forms with 52, 53 and 54 chromosomes do not show any differences in their reproductive rate. So a single autosome trivalent does not affect the fecondity. Many hybrids have been obtained betweenP. subterraneus (2N=52 or 54) andP. multiplex (2N=48) with 50 or 51 chromosomes, respectively. Even with a heterogenous karyotype they are not entirely sterile. 2 progeny were produced from back-crosses. The 2 parental species seem to result from recent speciation. Only fecondation or embryogenesis occurs in crossingP. duodecimcostatus withP. subterraneus orP. multiplex. The chromosome complement of 1 hybrid embryo ofP. duodecimcostatus (2N=62)×P. multiplex (2N=48) has been studied showing 55 chromosomes.P. duodecimcostatus appears to belong to a more distant evolutionary group.     

Explaining Leibniz equivalence as difference of non-inertial appearances: Dis-solution of the Hole Argument and physical individuation of point-events

“The last remnant of physical objectivity of space–time” is disclosed in the case of a continuous family of spatially non-compact models of general relativity (GR). The physical individuation of point-events is furnished by the autonomous degrees of freedom of the gravitational field (viz., the Dirac observables) which represent—as it were—the ontic part of the metric field. The physical role of the epistemic part (viz. the gauge variables) is likewise clarified as embodying the unavoidable non-inertial aspects of GR. At the end the philosophical import of the Hole Argument is substantially weakened and in fact the Argument itself dissolved, while a specific four-dimensional holistic and structuralist view of space–time (called point-structuralism) emerges, including elements common to the tradition of both substantivalism and relationism. The observables of our models undergo real temporal change: this gives new evidence to the fact that statements like the frozen-time character of evolution, as other ontological claims about GR, are model dependent.