Thai Acanthamoeba isolate (T4) induced apoptotic death in neuroblastoma cells via the Bax-mediated pathway

A Thai Acanthamoeba isolate named AS recovered from a corneal scraping of a keratitis patient was genotypically determined as T4. AS trophozoites were used for studying Acanthamoeba-induced apoptosis in mouse neuroblastoma NA cells during in vitro co-cultivation. The Acanthamoeba-exposed NA cells showed signs of apoptosis including cell shrinkage, nuclear condensation and DNA fragmentation. The effect was confirmed by DNA laddering electrophoresis. Involvement of caspase enzymes and mitochondrial pro- and anti-apoptotic proteins (Bax and Bcl-2) in AS-induced apoptosis was determined. The use of Z-VAD-FMK, a pan-caspase inhibitor, significantly reduced the apoptotic effect, while Bax/Bcl-2 ratio analysis showed a significant increase in the expression of apoptotic proteins in AS-exposed NA cells. These results strongly indicated that apoptosis induced by AS trophozoites is caspase-dependent and is mediated by over-expression of pro-apoptotic proteins in the mitochondrial pathway. This is the first report on the role of Bax in mediating apoptosis induced by Acanthamoeba.     

Production and characterization of monoclonal antibodies to Acanthamoeba castellanii and their application for detection of pathogenic Acanthamoeba spp

Fourteen monoclonal antibodies (mAbs) were produced against a strain of Acanthamoeba castellanii isolated from a human cornea. The reactivity of the mAbs to reference strains of Acanthamoeba was examined by an indirect fluorescence antibody test (IFA) and Western immunoblot analysis. Nine mAbs reacted specifically with a known pathogenic reference strain of A. castellanii, but not with a non-pathogenic strain or other Acanthamoeba spp. The antigen recognized by these mAbs had a molecular mass of 17 kDa. The remaining five mAbs reacted with A. castellanii and A. polyphaga, members of group II (Pussard and Pons) but not with A. astronyxis (group I) or A. culbertsoni (group III). Western immunoblot analysis revealed that the latter mAbs stained many protein bands ranging from 30 to 150 kDa. None of the 14 mAbs reacted with Naegleria gruberi, N. fowleri, or Entamoeba histolytica. These observations suggest that an antigen common in group II as well as a pathogenic A. castellanii-specific antigen are present. Slot blot reactivity was comparable to the IFA. Under certain circumstances, therefore, slot blot analysis with a panel of mAbs should be helpful in the detection of keratitis-producing strains of Acanthamoeba.     

Acanthamoeba affects the integrity of human brain microvascular endothelial cells and degrades the tight junction proteins

Haematogenous spread is a key step in the development of Acanthamoeba granulomatous encephalitis, however it is not clear how circulating amoebae cross the blood–brain barrier to enter the CNS to produce disease. Using the primary human brain microvascular endothelial cells (HBMEC), which constitute the blood–brain barrier, here it is shown that Acanthamoeba abolishes the HBMEC transendothelial electrical resistance. Using traversal assays, it was observed that Acanthamoeba crosses the HBMEC monolayers. The primary interactions of Acanthamoeba with the HBMEC resulted in increased protein tyrosine phosphorylations and the activation of RhoA, suggesting host–parasite cross-talk. Furthermore, Western blot assays revealed that Acanthamoeba degraded occludin and zonula occludens-1 proteins in a Rho kinase-dependent manner. Overall, these findings suggest that Acanthamoeba affects the integrity of the monolayer and traverses the HBMEC by targeting the tight junction proteins.     

文冠果有性生殖特征的观察研究

该研究以文冠果花发育不同时期采集的花药和胚珠为材料,采用光学及电子显微技术对花的性系统特征、胚珠发育及自交败育等有性生殖特征进行观察分析,以揭示文冠果开花多、结果少的内在机理。结果显示:(1)文冠果植物产生大量雄花和很少的两性花,表现隐形雌雄同株的性系统类型。(2)文冠果花拥有5个独特的金黄色角状附属物;成熟花粉粒球形,3孔沟,沟在花粉极面不相连,花粉表面有疣状纹饰;花主要以风媒传粉。(3)柱头3裂,开花时柱头表面布满丝状乳突细胞,属于干燥型。(4)卵球形弯生胚珠内有一弯曲的长胚囊,胚囊被厚珠心包围。胚珠受精后,珠孔与合点之间的珠柄侧外珠被的局部细胞平周和垂周分裂产生一个径向突起,伸向胚囊,与胚囊纵轴略成直角;受精后水和可溶性物质不断进入膨大的胚囊;幼胚的两子叶从胚囊珠孔端的管状部位进入合点端大的腔室中。(5)在胚发育早期,两片子叶是对称的,但在授粉35d后,具有较大半径的一子叶比另一子叶的体积大;授粉后23d以前,胚囊内的液态内容物含有游离核胚乳和少量细胞化胚乳,这个时期的胚珠可以鲜食。(6)自交传粉后,胚珠与幼果在合子形成后的不同时期败育。研究表明,文冠果与无患子科的其它植物有很多相似的有性生殖特征,但文冠果也具有一些异常特征,如花存在附属物及每子房室中有较多的胚珠等。     

蕨受精作用的细胞学观察

为了阐明进化蕨类受精作用的特点和细胞学机制,该文采用透射电镜观察了蕨(Pteridium aquilinum var.latiusculum)受精作用的主要过程,观察结果显示:(1)蕨精子通过受精孔进入卵细胞,多数情况下,该精子的螺旋运动先在受精孔的下方产生一个受精腔,然后精子再与卵细胞质融合。(2)第一个精子的这种延迟的螺旋运动和因精子的钻入而引起的卵细胞固缩反应可能是阻止多精受精的重要因素。(3)卵发育时期产生的核外突在受精后仍能持续12 h,然后与核本体分离,逐渐在细胞质中消解。(4)合子通过其后方细胞质的液泡化而建立了水平极性,此后再进行细胞分裂。该研究观察到了进化蕨类受精作用过程中的一些新现象,包括产生受精腔、卵细胞固缩反应、核外突的命运以及合子极性建立等,这有助于理解蕨类植物的受精作用机制及有性生殖的演化。     

Observations on the endogenous stages of Eimeria crandallis in domestic lambs (Ovis aries)

Lambs reared coccidia-free were inoculated orally with various numbers of sporulated oocysts of E. crandallis and were killed between 1 and 22 days after inoculation; tissues were examined histologically. Sporozoites were seen 1, 2 and 3 days after inoculation (DAI) in crypt epithelial cells in the mid-jejunum. Infected cells migrated into the lamina propria where the parasite within them developed into a firstgeneration meront containing about 250,000 merozoites at 10 DAI. A second generation of meronts was seen at 10–12 DAI, each containing up to about 10 merozoites, situated mainly at the bases of crypts in the jejunum and ileum but also in the caecum. From 11 DAI pro-gamonts were seen which were enveloped by the host cell nucleus and which divided in synchrony with the host cell for an undetermined number of generations. Mature gamonts began to develop from them by 16 DAI. Oocyst output began at 16 DAI and rose to a peak at about 22 DAI. Up to 10⁸ oocysts were produced per oocyst inoculated. They showed wide variation in size and colour.     

桃花水母生活史的实验观察

记录了中国产信阳桃花水母(Craspedacusta xinyangensis)的生活史及各阶段主要发育特征。25～28℃条件下,卵受精23h后发育成长0.14～0.21mm的圆棍状浮浪幼虫,养殖环境下,浮浪幼虫在水中漂浮3～5d后固定到人工玻璃底质上,3～4d后发育成长度0.3～0.6mm的螅状体。水螅体产生无纤毛的类浮浪幼虫形成新的螅状体。一个螅状体成熟后产生一个水母芽,新释放的幼水母具有16只触手。     

华东安蕨卵发生的超微结构观察

核心薄囊蕨是蕨类植物中的进化类群,但对受精作用具有显著影响的卵发生研究仍较少,该文利用超微技术对其中蹄盖蕨科的华东安蕨卵发生过程进行了研究,以进一步完善薄囊蕨植物卵发生的科学资料,为理解蕨类植物的有性生殖及演化机制奠定基础。超微结构观察显示:华东安蕨的幼卵和沟细胞在颈卵器中紧密联接;随后,在卵细胞上方出现了分离腔和临时细胞壁,但在卵细胞中间孔区处卵细胞和腹沟细胞始终联接在一起;分离腔中的无定形物质沉积在卵细胞的质膜外形成了1层加厚的卵膜,而在孔区处没有形成卵膜,该位置最后形成了受精孔。在进一步的卵发生过程中,卵细胞核变得高度不规则,形成了大量的核外突和核褶皱。     

Observations on Barnacle Goose Breeding,Branta leucopsis, in Spitsbergen 1975

Summary Our observations on Barnacle geese in Spitsbergen permit the following conclusions: The non-breeding geese molted at about the same time as hatching occurred in the nesting colonies, about 16 days before the breeders' molt began. The non-breeders came together into large groups along the coast. Pair bond displays (cackling and triumph ceremonies) were infrequent in non-breeder molting flocks. Families formed small flocks after hatching and migrating to the breeding areas. The flocks grew in number as the summer progressed and the parental molt occurred. The non-breeder flocks and the family flocks were both extremely cohesive. The families in a particular flock were normally synchronized in their activities. A flock of three families was studied in detail. An analysis of activity patterns showed that no diurnal fluctuations in resting, grazing and travelling were present. One gander initiated place changes more successfully than the other two. No geese or goslings were preyed upon during our observations. Gulls and skuas were threatened by individual geese. Arctic foxes caused flightless geese to take refuge in water. Full-winged adults mobbed the foxes.

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Zusammenfassung Unsere Beobachtungen an Nonnengänsen in Spitzbergen nach der Brutzeit brachten folgende Ergebnisse: Die Mauser setzte bei den nichtbrütenden Gänsen etwa 16 Tage früher ein als bei den brütenden und fiel zeitlich ungefähr mit dem Schlüpftermin der Gössel zusammen. Die Nichtbrüter bildeten große Mauserverbände in Küstennähe und unterließen fast völlig Verhaltensweisen, die dem Paarzusammenhalt dienen (Schnattern und Triumphgeschrei). Die Brüter wanderten nach dem Schlüpfen der Jungen von den Brutplätzen zu ihren Aufzuchtgebieten, wo sie sich zu kleineren Familienscharen zusammenschlossen. Diese Familienscharen nahmen im Lauf des Sommers und mit dem Einsetzen der elterlichen Mauser zahlenmäßig zu. Sowohl in den Mauser- als auch in den Familienscharen zeigten die Individuen einen starken Zusammenhalt auf engem Raum und synchrone Aktivitäten. Eine Dreifamilienschar wurde im Detail untersucht. Ein Tagesrhythmus in der zeitlichen Verteilung von Weiden, Ruhen und Ortswechsel trat nicht auf. Einem Ganter gelang es häufiger als den beiden anderen, bei Ortsveränderungen eine führende Rolle in der Schar einzunehmen. Die Ganter richteten ihre Angriffe bevorzugt auf andere Ganter und fremde Gössel, die Weibchen auf andere Weibchen. Wir konnten nicht beobachten, daß Gänse oder Gössel von Raubtieren erbeutet wurden. Adulte Gänse griffen Möwen und Raubmöwen an. Vor einem Eisfuchs begaben sich flugunfähige Gänse ins Wasser, während flugfähige Gänse ihn anhaßten.

     

Ecdysterone induced sexual reproduction in Opalina sudafricana parasitic in Bufo regularis

Ecdysterone induced sexual reproduction in Opalina sudafricana parasitic in Bufo regularis. International Journal for Parasitology 3: 863–868. Ecdysterone injections (0·5 mg/ml) induced sexual reproduction in Opalina sudafricana parasitic in male or female Bufo regularis after 12 days. Hypophysectomy or gonadectomy did not prevent the hormone from producing its effect on the parasites. Ecdysterone did not induce sexual reproduction in the parasites in vitro. Urine of toads injected with ecdysterone induced the opalinids to encyst in vitro.     

Observations on the cross-immunity between Theileria lawrencei (Serengeti) and Theileria parva (Muguga) in cattle

Observations on the cross-immunity between Theileria lawrencei (Serengeti) and Theileria parva (Muguga) in cattle. Internationaljournal for Parasitology 3: 723–728. Cattle immunized against Theileria parva (Muguga) showed little resistance to Theileria lawrencei (Serengeti) stabilate challenge, while cattle immune to T. lawrencei (Serengeti) were fully resistant to challenge with T. parva (Muguga) stabilate. Cattle inoculated with cultured lymphoid cells infected with T. lawrencei (Serengeti) macroschizonts survived a subsequent T. lawrencei (Serengeti) stabilate challenge.     

Y chromosome-specific mutations induced by a giant transposon in Drosophila hydei

Summary The w ^m Co duplication of Drosophila hydei (Dp (1; Y) 16B2-17B1) contains 13–16 bands in salivary gland chromosomes. The duplication resides preferentially in the X heterochromatin or on the Y chromosome. In some stocks frequent (up to 4×10^-3) exchanges of the duplication occur between different Y chromosomes (T(X; Y) and free Y) or between the X and the Y chromosome. About 60% of the T(X; Y)-Y exchanges induce mutations in the Y chromosomal male fertility genes of the recipient Y chromosome. From the mutational spectrum generated by the T(X; Y)-Y transpositions and from the variable efficiency as acceptor of different X-Y translocations it can be concluded that the exchanges show a remarkable site specificity: distal positions in the long arm of the Y chromosome are occupied preferentially. More proximal positions in the long arm of insertions into the short arm of the Y chromosome are found only with a lower frequency. No transpositions to the autosomes have been recovered. Duplications are lost with highly differing frequencies. The losses are not linked with insertions of the w ^m Co element into a new position and are more frequent than transpositions. Therefore, we regard the w ^m Co element as a giant transposon.