Morphological changes in trypanosoma brucei rhodesiense following inhibition of polyamine biosynthesis in vivo

The effect of α-difluoromethylornithine (DFMO) treatment on the morphology of African trypanosomes was investigated. For this purpose inbred mice were immunosuppressed and infected with a clone of the protozoan blood parasite Trypanosoma brucei rhodesiense. The mice were then treated with DFMO, an irreversible inhibitor of ornithine decarboxylase, which inhibits polyamine synthesis. DFMO treatment in the absence of host immunity resulted in arrest of cytokinesis of the trypanosomes and many binucleated cells could be seen in blood smears. If mice were infected with a highly virulent trypanosome clone (ETat 1.10), which does not normally transform from long slender (LS) to short stumpy (SS) forms, DFMO treatment caused SS transformation to occur on days 3–4. This morphological SS transformation was substantiated by the presence of diaphorase activity and nuclear and mitochondrial changes. The results suggest a possible involvement of polyamines in the transformation from LS to SS forms.     

Oligopeptide Signaling through TbGPR89 Drives Trypanosome Quorum Sensing

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菠菜叶片中两种酶型的磷酸果糖激酶性质的比较

菠菜叶片提取液经ＰＥＧ－６０００沉淀、ＤＥ－５２离子交换柱层析及分子筛ＳｅｐｈｅｃｔｙｌＳ－３００凝胶过滤得到两种分子量不同的依赖ＡＴＰ的磷酸果糖激酶（ＰＦＫ）。一为大分子酸型,分子量大于２０００ｋＤ,其活力可被Ｐｉ、３－ＰＧＡ、柠檬酸激活,被ＰＥＰ强烈抑制,Ｐｉ能减缓此抑制作用,Ｍｇ２＋为必需金属离子,但其浓度高于０．５ｍｍｏｌ／Ｌ时酶活力降低;一为小分子酸型,分子量为３００ｋＤ,其活性受Ｐｉ、３－ＰＧＡ、柠檬酸和ＰＥＰ抑制,Ｍｇ２＋亦为必需金属离子,Ｈｉｌｌ系数为０．６７,表现负协同效应。实验证明小分子酸型可能存在叶绿体中,大分子酸型属于胞质酶。     

Studies of Blastocystis hominis

Blastocystis hominis, grown in Boeck-Drbohlav culture medium, modified by the omission of rice starch and the addition of 20% human serum and mineral oil cover to the Locke's solution overlay, can assume 3 morphologic forms. In the absence of human serum the vacuolated form, which divides by binary fission predominates. In medium with high serum content the granular form appears, with 3 types of granules. Spheroid or more elongate cytoplasmic granules predominate. In older organisms, lipid granules are found either in the peripheral cytoplasm or in the central vacuolar space. In occasional cells, variable numbers of reproductive granules develop in the central vacuolar space. These latter granules are released from the organism and give rise to typical B. hominis cells. The 3rd form, the ameba form, appears in small numbers in older cultures and in those treated with antibiotics. Ameba forms feed on bacteria and have slow pseudopodial activity. Exposure to oxygen causes rapid damage to cell membrane, with resulting leakage and collapse.     

Survival and Infectivity of Pneumocystis carinii Outside the Mammalian Host.

SUMMARY Air blower prefilters servicing HEPA-filtered Biobubble housing Pneumocystis carinii -infected rats were stored for up to five months at -80°C to room temperature. After storage, 76% of immunosuppressed rats exposed to these prefilters developed P. carinii infections. In contrast, only 4% of control immunosuppressed rats exposed to autoclaved filters had P. carinii infections. These observations indicate that the organism has a dormant form that remains infective for at least several months outside the mammalian host.     

Protein Uptake and Digestion in Bloodstream and Culture Forms of Trypanosoma brucei*

SYNOPSIS. The mechanisms of ferritin uptake and digestion differ in bloodstream and culture forms of Trypanosoma brucei. Ferritin enters bloodstream forms from the flagellar pocket by pinocytosis in large spiny-coated vesicles. These vesicles become continuous with straight tubular extensions of a complex, mostly tubular, collecting membrane system where ferritin is concentrated. From the collecting membrane system the tracer enters large digestive vacuoles. Small spiny-coated vesicles, which never contain ferritin, are found in the Golgi region, fusing with the collecting membrane system, and around the flagellar pocket. Acid phosphatase activity is present in some small spiny-coated vesicles which may represent primary lysosomes. This enzymic activity is also found in the flagellar pocket, pinocytotic vesicles, the collecting membrane system, the Golgi (mature face), and digestive vacuoles of bloodstream forms. About 50% of the acid phosphatase activity of blood forms is latent. The remaining nonlatent activity is firmly cell-associated and probably represents activity in the flagellar pocket. The structures involved in ferritin uptake and digestion are larger and more active in the short stumpy than in the long slender bloodstream forms. The short stumpy forms also have more autophagic vacuoles. No pinocytotic large, spiny-coated vesicles or Golgi-derived, small spiny-coated vesicles are seen in culture forms. Ferritin leaves the flagellar pocket of these forms and enters small smooth cisternae located just beneath bulges in the pocket membrane. The tracer then passes through a cisternal collecting membrane network, where it is concentrated, and then into multivesicular bodies. In the culture forms, acid phosphatase activity is localized in the cisternal system, multivesicular bodies, the Golgi (mature face), and small vesicles in the Golgi and cisternal regions. The flagellar pocket has no acid phosphatase activity, and almost all the activity is latent in these forms. The culture forms do not release acid phosphatase into culture medium during 4 days growth. Uptake of ferritin by all forms is almost completely inhibited by low temperature. These differences among the long slender and short stumpy bloodstream forms and culture forms are undoubtedly adaptive and reflect different needs of the parasite in different life cycle stages.     

记澜沧江河谷一硬叶小叶常绿阔叶林类型——锈鳞木樨榄、清香木群丛

采用法瑞地植物学派的样地调查法记录了澜沧江中上游河谷中一独特的硬叶小叶常绿阔叶林类型——锈鳞木樨榄、清香木群丛。该群丛由南向北分布于1300~1900m的河谷下部,对群落外貌、生活型与生长型、植物种类科属的组成、分布区类型等进行了分析。结果表明：（1） 群落结构简单,物种数平均为18种;（2） 生活型以高位芽植物居多,生长型以草本居多;（3） 群落中计有维管植物116种,隶属于39科,84属;（4） 植物科级、属级水平上均显示出了热带性质,植物区系表现出一定的古老性,反映了与古地中海硬叶小叶林的历史渊源。该植物群丛适应澜沧江干热-干暖河谷气候,分布区域狭窄,是一类值得重点保护和深入研究的植被类型。     

The obesity in bilateral ovariectomized rats is related to a decrease in the expression of leptin receptors in the brain.

We investigated the expression levels of leptin receptors in the brain of ovariectomized (OVX) rats. The mean expression level of ob mRNA in adipose tissues of OVX rats was significantly (P < 0.01) lower than that in the SHAM operation group rats, and the mean body weight of OVX rats was significantly (P < 0.01) greater than that in the SHAM group rats. However, there were no differences between serum leptin concentrations in these two groups. The mean level of leptin receptor (OB-R) mRNA expression in the brain tissue and the mean level of long form type OB-R (OB-RL) mRNA expression in the hypothalamus of the OVX rats were significantly (P < 0.05) lower than those in the SHAM group rats. These changes were cancelled by supplementation with 17 beta-estradiol in OVX rats. These results suggested that not only changes in the expression level of ob mRNA in adipose tissue and the serum leptin concentration but also changes in the OB-R mRNA in the brain are involved in the body weight increase in OVX rats and that a decrease in OB-R makes transmission of signals to suppress the amount of food intake difficult, thus leading to an increase in body weight.     

云南山楂茎尖的离体培养及其无性系快速繁殖

用云南山楂(Crataegus scabrifolia(Franch.)Rehd.)成年树茎尖和实生芽两种不同发育阶段的材料为外殖体,诱导它们休眠芽萌动,丛生芽条并诱导芽条生根。实验结果如下:1.以成年态的云南山楂侧芽为外植体,培养在附加IAA 0.1—0.5mg/l+6-BA 1-2mg/l的MS培养基上可诱导芽的萌发;将芽继代培养在附加0.5—1mg/l 6-BA的SH或MS培养基上,40天后芽数增殖4—6倍;将芽条截下置于1/2MS培养基上,附加不同浓度的IAA或IBA,可得到50—80％的生根率。2.以实生芽为外殖体,在相同条件下,则20天后芽数增殖便可获4—6倍;98％以上生根。结果表明:云南山楂的幼年态要比成年态易脱分化和再分化。     

Peptides derived from pyruvate dehydrogenase as substrates for pyruvate dehydrogenase kinase and phosphatase.

Evidence is presented that phosphopeptides produced by tryptic digestion of phosphorylated pyruvate dehydrogenase are effective substrates for pyruvate dehydrogenase phosphatase and that the dephosphopeptides can serve as substrates for pyruvate dehydrogenase kinase. These findings indicate that the phosphatase and the kinase do not require an intact tertiary structure in pyruvate dehydrogenase, but apparently recognize components of the local primary sequence around the phosphorylation sites.     

A floristic-structural gradient related to land forms in the southern Chihuahuan Desert

Abstract. The purposes of this study were to elucidate the floristic and structural characteristics of the vegetationin the Mapimi subdivision of the Chihuahuan Desert, Mexico, and to relate them to environmental variation. The main question addressed was: How does floristic composition, total species richness and life-form species richness vary in relation to environmental change? 154 sites, randomly selected and stratified over seven landscape units, were analyzed. Results showed the existence of a land form gradient along which vegetation types were ordered. Species richness varied along the gradient, the richest land form was bajada, indicating that the maximum species richness did not occur at one extreme of the water availability gradient but in a moderately poor situation. The lowest species richness was found in the playa land form. Cover-based life form spectra varied significantly with land forms, while presence-absence based life form spectra did not. It is suggested that this may be a consequence of the relatively young age of this desert.     

土壤中重金属形态分析及其环境学意义

介绍了土壤重金属的形态及各种分析方法,重点说明了土壤中重金属形态分布及影响因素;讨论了影响土壤环境中重金属形态转化的因素,重金属形态与重金属在土壤中的迁移性、可给性、活性的关系,重金属污染土壤修复与重金属形态分布的关系.形态分析在一定程度上反映自然与人为作用对土壤中重金属来源的贡献,并反映重金属的生物毒性.重金属可以因形态中某一个或几个方面不同而表现出不同的毒性和环境行为.     

Lack of repair of ultraviolet light damage in Mycoplasma gallisepticum

Molecules with single-stranded tails (rolling circles) were isolated as replicating intermediates in G4 progeny single-stranded DNA synthesis. Lysates from infected cells harvested late in infection during single-stranded DNA synthesis were not deproteinised but analysed directly in caesium chloride and propidium diiodide gradients. The gradient fractionated them on the basis of tail length. If the lysates were first deproteinised however, the tailed replicative intermediates banded as a peak at a density just greater than that of replicative form II DNA (RFII) and did not spread down the gradient. The origin of synthesis of the viral strand tail was mapped by electron microscopy as 55 to 60% away from the single EcoRI cleavage site. Termination molecules finishing a round of viral strand DNA synthesis have been identified as molecules consisting of a closed single-stranded DNA circle attached by a very small region to the parent double-stranded DNA circle.     

Comparison of partial denaturation maps with the known sequence of simian virus 40 and phi X174 replicative form DNA.

Electron microscope partial denaturation maps of two viral DNAs, simian virus 40 and φX174 replicative form, have been obtained. A simple computer program has been developed to predict denaturation maps from any given DNA sequence, based on the percentage of A · T base-pairs along the molecule. Maps constructed from the SV40 DNA and φX174 replicative form DNA base sequence show a good correlation with the experimental maps. The results show that the regions of a DNA molecule that denature first are, in fact, those regions with the highest content of adenine and thymine base-pairs.     

Transcriptional organization of the Drosophila melanogaster ribosomal RNA genes.

Five distinct DNA replicating intermediates have been separated from lysates of bacteriophage G4-infected cells pulse-labelled during the period of replicative form synthesis using propidium diiodide/caesium chloride gradients. These are a partially single-stranded theta structure that is labelled in both the viral and complementary DNA strands; partially single-stranded circles, some with an unfinished viral DNA strand (25%) and some with an unfinished complementary DNA strand (75%); replicative form II(RFII) and replicative form I(RFI) DNA labelled only in the complementary DNA strand. To explain the pulse-label data a model is proposed in which G4 replicative form replication takes place by a displacement mechanism in which synthesis of the new viral DNA strand displaces the old viral DNA strand as a single-stranded DNA loop (D-loop) and when the displacement reaches half way round the molecule (the origin of synthesis of the G4 viral and complementary DNA strands are on opposite sides of the genome, Martin &; Godson 1977) synthesis of the complementary DNA strand starts, but in the opposite direction. Strand separation of the parent helix runs ahead of DNA synthesis, releasing two partially single-stranded circles from the replicating structure which then complete their replication as free single-stranded DNA circles. No evidence was found to support a rolling circle displacement mechanism of G4 replicative form synthesis.     

G4 DNA replication: I. Origin of synthesis of the viral and complementary DNA strands

The break in the complementary DNA strand of early G4 replicative form II DNA (RFII) and in the viral DNA strand of late RFII DNA was located using two single cleavage restriction enzymes (EcoRI and PstI) and by limited nick translation of the break using DNA polymerase I and ³²P-labelled deoxyribonucleotides followed by digestion with the restriction enzymes HaeIII and HindII. The break in the complementary DNA strand was unique and in HaeIII Z5 close to the EcoRI cleavage site whereas the break in the viral DNA strand was on the other side of the molecule in HaeIII Z2 approxiately 50% away from the EcoRI cleavage site. Distribution of a short ³H pulse in early G4 replicating intermediates that were synthesising both DNA strands at the same time showed that synthesis of the strands started on opposite sides of the molecule and proceeded in opposite convergent directions, suggesting that initiation of synthesis of the two strands was independent and not unified in a single growing fork.     

幽门螺杆菌球形体回复原形的实验研究

目的对幽门螺杆菌（Helicobacter pylori,Hp）球形体进行体内、外回复原形的比较研究,揭示其潜在的传播途径。方法在布氏肉汤的基础上设计了4种Hp再生培养基,对Hp螺旋体、原生质体及球形体进行体外培养;同时采用30只蒙古沙土鼠（Mongolian gerbil）进行体内感染定植实验,对感染小鼠胃粘膜进行Hp定量培养和组织学检测。结果Hp球形体在4种再生培养基中均未能回复生长;而在感染小鼠的体内却观察到了Hp球形体的回复定植。Hp螺旋体感染组在小鼠胃内的定植密度较高,且胃粘膜下可见大量炎症细胞浸润;而球形体感染组并未见到小鼠胃粘膜组织的明显炎症损伤,且仅有少量回复的螺旋体定植。结论Hp球形体作为一种低水平代谢休眠体,代谢活性及毒力均有所减弱,但仍具有潜在的致病性。本研究支持部分Hp球形体具有活力但体外不能培养成活这一假说,提示＂粪-口＂传播应引起更多的关注。     

A rolling circle model of the in vivo replication of bacteriophage phiX174 replicative form DNA: different fate of two types of progeny replicative form

In a preceding paper (Schröder and Kaerner, 1972) a rolling circle mechanism has been described for the replication of bacteriophage φX174 replicative form. Replication involved nicking and elongation of the viral (positive) strand component of the RF molecule resulting in the displacement of a single-strand tail of increasing length. The synthesis of the new complementary (negative) strand on the single-strand tails appears to be initiated with considerable delay and converts the tail into double-stranded DNA. Before the new negative strand is completed the replicative intermediates split into (I) a complete RF molecule containing the “old” negative and the new positive strand, and (II) a linear, partially double-stranded “tail” consisting of the complete old positive strand and a fragment of the new negative strand.The present study is concerned with the fate during RF replication of these fragments of the rolling circles. Those RFII molecules containing the old negative strands appear to go into further replication rounds repeatedly. Some of the tails were found in the infected cells in their original linear form. “Gapped” RFII molecules, which have been described earlier by Schekman and co-workers (Schekman &; Ray, 1971; Schekman et al., 1971), are supposed to originate from the tails of rolling circle intermediates by circularization of their positive strand components. Evidence is provided by our experiments that even late during RF replication these gaps are present only in the negative strands of RFII. Appropriate chase experiments indicated that the tails finally are converted to RFI molecules. Progeny RFI molecules could not be observed to start new replication rounds under our conditions although we cannot exclude that this might happen to some minor extent.The results presented suggest that the master templates for RF replication are the first negative strands to be formed, rather than the parental positive strands.     

淡黄金花茶一新变型

<正> 本变型与原变型淡黄金花茶主要区别在于花瓣多达23—30片,长圆形或条形,其中9—14片淡红色,14—20片淡黄色;雄蕊188枚,其中花丝95枚特粗,子房极小。花期11月。