The glio-toxic mechanism of α-aminoadipic acid on cultured astrocytes

The mechanism of action of the glutamate analogue α-aminoadipic (A A A) acid was investigated in terms of its toxicity to cultured astrocytes. A A A was more toxic to type 1 astrocytes than type 2 astrocytes. Also the higher toxicity of the L-isomer as compared to the D-isomer was seen on type 1 astrocytes but not type 2. The toxicity of A A A can be reduced by co-culture of type 1 astrocytes with microglia. This inhibition may be due to glutamate release by microglia. No such effect is seen for type 2 astrocytes. The major uptake route for A A A by type 1 astrocytes is through the sodium dependent glutamate port. Both isomers of A A A are toxic to dividing astrocytes. The D-isomer appears to be toxic only for mitotic cells. The mechanism of this toxicity is protein synthesis dependent. It is suggested that A A A is toxic to mitotic astrocytes by interference with protein synthesis needed for cell division. D-A A A as opposed to L-A A A may prove a valuable tool for investigation of astrocyte proliferation in development and disease.     

A revision of the genus Annesorhiza (Apiaceae)

The genus Annesorhiza is revised and twelve species are recognized, of which two are newly described: A. altiscapa, A. burttii, A. flbrosa, A. flagellifolia, A. grandiflora, A. lateriflora, A. latifolia, A. macrocarpa, A. nuda, A. schlechteri, A. thunbergii and A. wilmsii. A. marlothii is reduced to synonymy under A. lateriflora , a species hitherto poorly known as Peucedanum lateriflorum. A. thunbergii is only known from the type specimen. A. elata, A. hirsuta and A. villosa are sunk under A. grandiflora. A. fúicaulis has recently been excluded from Annesorhiza on the basis of fruit structure. A key to the species is provided, an update on the nomenclature, typification of names and distribution maps are provided for all the species.     

Three new species of Aiphanes (Palmae) with notes on the genus in Ecuador

Twelve species of the palm genus Aiphanes occur in Ecuador. The morphological variation in the genus is surveyed, and the distribution of the Ecuadorean species is discussed. A key to the species of Aiphanes in Ecuador is provided. Aiphanes chiribogensis, A. grandis , and A. verrucosa are described as new species, and illustrated. Aiphanes caryotifolia, A. eggersii, A. erinacea, A. fosteriorum, A. gela-tinosa, A. macroloba, A. schultzeana , and A. tricuspidata are characterized. One still unidentified species resembling A. tessmannii is discussed.     

Systematic studies on the Pompilidae occurring in Japan: Genus Agenioideus Ashmead (Hymenoptera), supplement

Two species of the pompilid genus Agenioideus occurring in Japan are described: A . ( Agenioideus ) kokyo and A . ( A .) cinctellus . The former is new to science and the latter is recorded from Japan and Taiwan for the first time. A brief summary of the biology of A. cinctellus , distinguishing characters between A. ( A. ) ishikawai Shimizu, 1989 and its relatives, and a key to all Japanese species of this genus are presented. Psammochares cinctellus f. rufa Haupt, 1938 and A . ( A. ) pacificus Lelej, 1994 are newly synonymized with A. cinctellus . A new combination is proposed: A . ( A .) maculipes (Smith, 1870) (= Pompilus maculipes Smith), which is found in Southeast Asia to South Asia. Agenioideus ishikawai is newly recorded from Korea and China.     

Isoprenylation is required for the processing of the lamin A precursor

The nuclear lamina proteins, prelamin A, lamin B, and a 70-kD lamina-associated protein, are posttranslationally modified by a metabolite derived from mevalonate. This modification can be inhibited by treatment with (3-R,S)-3-fluoromevalonate, demonstrating that it is isoprenoid in nature. We have examined the association between isoprenoid metabolism and processing of the lamin A precursor in human and hamster cells. Inhibition of 3-hydroxy-3-methylglutaryl coenzyme A reductase by mevinolin (lovastatin) specifically depletes endogenous isoprenoid pools and inhibits the conversion of prelamin A to lamin A. Prelamin A processing is also blocked by mevalonate starvation of Mev-1, a CHO cell line auxotrophic for mevalonate. Moreover, inhibition of prelamin A processing by mevinolin treatment is rapidly reversed by the addition of exogenous mevalonate. Processing of prelamin A is, therefore, dependent on isoprenoid metabolism. Analysis of the conversion of prelamin A to lamin A by two independent methods, immunoprecipitation and two-dimensional nonequilibrium pH gel electrophoresis, demonstrates that a precursor-product relationship exists between prelamin A and lamin A. Analysis of R,S-[5-3H(N)]mevalonate-labeled cells shows that the rate of turnover of the isoprenoid group from prelamin A is comparable to the rate of conversion of prelamin A to lamin A. These results suggest that during the proteolytic maturation of prelamin A, the isoprenylated moiety is lost. A significant difference between prelamin A processing, and that of p21ras and the B-type lamins that undergo isoprenylation-dependent proteolytic maturation, is that the mature form of lamin A is no longer isoprenylated.     

Non-snRNP U1A levels decrease during mammalian B-cell differentiation and release the IgM secretory poly(A) site from repression

A regulated shift from the production of membrane to secretory forms of Immunoglobulin M (IgM) mRNA occurs during B cell differentiation due to the activation of an upstream secretory poly(A) site. U1A plays a key role in inhibiting the expression of the secretory poly(A) site by inhibiting both cleavage at the poly(A) site and subsequent poly(A) tail addition. However, how the inhibitory effect of U1A is alleviated in differentiated cells, which express the secretory poly(A) site, is not known. Using B cell lines representing different stages of B cell differentiation, we show that the amount of U1A available to inhibit the secretory poly(A) site is reduced in differentiated cells. Undifferentiated B cells have more total U1A than differentiated cells and a greater proportion of this is not associated with the U1snRNP. We show that this is available to inhibit poly(A) addition at the secretory poly(A) site using cold competitor RNA oligos to de-repress poly(A) addition in nuclear extracts from the respective cell lines. In addition, endogenous non-snRNP associated U1A-immunopurified from the different cell lines-inhibits poly(A) polymerase activity proportional to U1A recovered, suggesting that available U1A level alone is responsible for changes in its inhibitory effect at the secretory IgM poly (A) site.     

中国黄耆属植物一些种类的订正

在对中国黄耆属植物进行研究的过程中,发现一些不同时期发表的类群存在同物异名现象。对这些类群进行了归并,将Astragalus dulanensis Y．H．Wu作为A．variabilis Bunge的新异名,A．hepingensis Liou f和A．mongutensis Lipsky作为A．kuschakewiczi B．Fedtsch ex O．Fedtsch的新异名,A．zhaolingicus K．T．Fu作为A．galactites Pall．的新异名,A．xipingshanicus Y．H．wu作为A．minshanensis K．T．Fu的新异名,A．transiliensis Gontsch.var．microphyllus S．B．Ho作为A．ochrias Bunge的新异名。     

Analysis of β3-endonexin mutants for their ability to interact with cyclin A

We have recently identified beta(3)-endonexin as a molecule that interacts with cyclin A-associated kinase. In this study, beta(3)-endonexin mutants were constructed by PCR-based site-directed mutagenesis, and characterized. Beta(3)-endonexin has a cyclin binding motif, RxL, in its N-terminal region, and two SP sequences which resemble a known target site for cyclin-dependent kinases (Cdks). The R5A/L7A mutant of beta(3)-endonexin, in which the RxL motif has been changed to AxA, is unable to bind to cyclin A, as revealed by two-hybrid experiments and in vitro pull-down assays. A GST-beta(3)-endonexin fusion, but not the corresponding R5A/L7A mutant, inhibits phosphorylation of Rb protein by cyclin A/Cdk2 in vitro. A cyclin A/Cdk2 kinase complex produced in, and purified from, insect cells phosphorylated GST-beta(3)-endonexin in vitro. The S33A or S46A mutant is partially phosphorylated by cyclin A/Cdk2, whereas no phosphorylation of the S33A/S46A double mutant is detectable. This demonstrates that these two serine residues, each of which is followed by a proline residue, are target sites for phosphorylation by cyclin A-associated kinase. The R5A/L7A mutant form of beta(3)-endonexin, which is defective for binding to cyclin A, is also not phosphorylated by cyclin A/Cdk2, confirming that the phosphorylation requires binding to cyclin A in the kinase complex. The neutralizing effect of beta(3)-endonexin on the toxicity associated with the expression of full-length human cyclin A in budding yeast is correlated with its ability to bind to cyclin A. Taken together, these data suggest that beta(3)-endonexin is phosphorylated by cyclinA/Cdk2 in vitro and that cyclin A-associated kinase activity is inhibited by the binding of beta(3)-endonexin to the kinase complex.     

Probing the local conformational change of alpha1-antitrypsin

The native form of serpins (serine protease inhibitors) is a metastable conformation, which converts into a more stable form upon complex formation with a target protease. It has been suggested that movement of helix-F (hF) and the following loop connecting to strand 3 of beta-sheet A (thFs3A) is critical for such conformational change. Despite many speculations inferred from analysis of the serpin structure itself, direct experimental evidence for the mobilization of hF/thFs3A during the inhibition process is lacking. To probe the mechanistic role of hF and thFs3A during protease inhibition, a disulfide bond was engineered in alpha(1)-antitrypsin, which would lock the displacement of thFs3A from beta-sheet A. We measured the inhibitory activity of each disulfide-locked mutant and its heat stability against loop-sheet polymerization. Presence of a disulfide between thFs3A and s5A but not between thFs3A and s3A caused loss of the inhibitory activity, suggesting that displacement of hF/thFs3A from strand 5A but not from strand 3A is required during the inhibition process. While showing little influence on the inhibitory activity, the disulfide between thFs3A and s3A retarded loop-sheet polymerization significantly. This successful protein engineering of alpha(1)-antitrypsin is expected to be of value in clinical applications. Based on our current studies, we propose that the reactive-site loop of a serpin glides through between s5A and thFs3A for the full insertion into beta-sheet A while a substantial portion of the interactions between hF and s3A is kept intact.     

12种中国葱属植物的核型分析

采用细胞压片法,对采自中国西部的12种葱属植物的根尖有丝分裂中期进行了观察,其巾天蓝韭(A.cyaneum)、梭沙韭(Aforrestii)、昌都韭(A.changduense)、西川非(A.xichuanense)、野黄韭(A.rude)、野葱(A.chrysanthum)和真籽韭(A.eusperma)等7种植物的核型为首次报道.供试类群中,峨眉韭(A.omeiense)和多星韭(A.wallichii)的染色体基数分别为11和7,其余类群的染色体基数均为8.观察发现,随体杂合和多侪性现象在供试类群中很普遍.分析推测:(1)随体和倍性的变异在葱属某些类群的进化中可能起重要作用,随体的类型在葱属具有重要的分类意义;(2)多倍化和地下走茎的无性繁殖方式可能是天蓝韭(A.cyaneum)的进化策略;(3)西川韭(A.xichuanense)、野黄非(A.rude)和野葱(A.chrysanthum)有密切的亲缘关系;(4)真籽韭(A.eusperma)与多籽组在核型上有密切的亲缘关系.     

中国东北科尔沁沙地两种建群植物的抗旱机理

在土壤田间持水和干旱胁迫条件下,对冷蒿和差巴嘎蒿的多个抗旱性生理指标进行了测定,结果表明:1、在两种土壤水分状况下,冷蒿的水势低于差巴嘎蒿,水分相对亏缺、束缚水含量、束缚水与自由水比值、综合抗旱性指数均明显高于差巴嘎蒿,且胁迫前后冷蒿上述指标的变化幅度高于差巴嘎蒿。2、土壤田间持水量条件下冷蒿干物质累积量、硝态氮含量和硝酸还原酶活性高于差巴嘎蒿,可溶性蛋白质含量和叶绿素含量低于差巴嘎蒿。水分胁迫发生时,冷蒿蛋白质降解的幅度高于差嘎蒿;冷蒿体内脯氨酸的累积量可达胁迫前的8.2倍,差巴嘎蒿只达原来的2.2倍;冷蒿可溶性糖含量达胁迫前的1.29倍,差巴嘎蒿只达胁迫前的0.37倍;胁迫条件下冷蒿的叶绿素含量高于差巴嘎蒿。3、严酷的沙区环境条件下,冷蒿在一日内各时刻的蒸腾速率和水势均低于差巴嘎蒿,且日间波动平缓。     

横断山区四种湍蛙的细胞遗传学研究

通过染色体组型分析,C带(BSG技术)分析及一种简便的Ag-NORs带分析,对四川湍蛙、理县湍蛙、棕点湍蛙和棘皮湍蛙的种间关系、染色体的演化及其性染色体等问题进行了初步探讨。结果表明:(1)四川湍蛙、理县湍蛙和棕点湍蛙之间的亲缘关系较近,而它们与棘皮湍蛙的亲缘关系较远;(2)在近缘种的分化中,染色体结构异染色质的变化和臂间倒位是重要的因素之一,这在小型染色体上表现得尤为突出;(三)四川湍蛙具有在形态上分化很明显的性染色体。C带分析表明,此性染色体主要由常染色质构成,但在其Y染色体的长臂上存在明显的中间C带,推测尚处于性染色体分化的初期阶段。     

The Mutation SK(ad-3A) Cancels the Dominance of ad-3A+ over ad-3A in the Ascus of Neurospora

A newly induced mutant of Neurospora, when crossed with an ad-3A mutant, produces asci with four viable black and four inviable white ascospores. The survivors always contain the new mutant allele, never ad-3A. The new allele, which is called SK(ad-3A) (for spore killer of ad-3A), is located at or very near the ad-3A locus.--In crosses homozygous for ad-3A, each ascus contains only inviable white ascospores. This defect in ascospore maturation is complemented by the wild-type allele, ad-3A+ (crosses heterozygous for ad-3A and ad-3A+ produce mainly viable ascospores), but it is not complemented by the new SK(ad-3A) allele (all ad-3A ascospores from crosses heterozygous for SK(ad-3A) and ad-3A are white and inviable). In crosses homozygous for SK(ad-3A) or heterozygous for SK(ad-3A) and ad-3A+, each ascus contains only viable black ascospores. SK(ad-3A) does not require adenine for growth, and forced heterokaryons between SK(ad-3A) and ad-3A grow at wild-type rates and produce conidia of both genotypes with approximately equal frequency. Thus, the action of SK(ad-3A) is apparently restricted to ascospore formation. Possible mechanisms of the action of this new allele are discussed.     

Decavanadate inhibits catalysis by ribonuclease A

Pentavalent organo-vanadates have been used extensively to mimic the transition state of phosphoryl group transfer reactions. Here, decavanadate (V(10)O(28)6-) is shown to be an inhibitor of catalysis by bovine pancreatic ribonuclease A (RNase A). Isothermal titration calorimetry shows that the Kd for the RNase A decavanadate complex is 1.4 microM. This value is consistent with kinetic measurements of the inhibition of enzymatic catalysis. The interaction between RNase A and decavanadate has a coulombic component, as the affinity for decavanadate is diminished by NaCl and binding is weaker to variant enzymes in which one (K41A RNase A) or three (K7A/R10A/K66A RNase A) of the cationic residues near the active site have been replaced with alanine. Decavanadate is thus the first oxometalate to be identified as an inhibitor of catalysis by a ribonuclease. Surprisingly, decavanadate binds to RNase A with an affinity similar to that of the pentavalent organo-vanadate, uridine 2',3'-cyclic vanadate.     

Alzheimer beta-amyloid peptides: normal and abnormal localization

Alzheimer's disease (AD) neuropathology is characterized by accumulation of "senile" plaques (SPs) and neurofibrillary tangles (NFTs) in vulnerable brain regions. SPs are principally composed of aggregates of up to 42/43 amino acid beta-amyloid (A beta) peptides. The discovery of familial AD (FAD) mutations in the genes for the amyloid precursor protein (APP) and presenilins (PSs), all of which increase A beta42 production, support the view that A beta is centrally involved in the pathogenesis of AD. A beta42 aggregates readily, and is thought to seed the formation of fibrils, which then act as templates for plaque formation. A beta is generated by the sequential intracellular cleavage of APP by beta-secretase to generate the N-terminal end of A beta, and intramembranous cleavage by gamma-secretase to generate the C-terminal end. Cell biological studies have demonstrated that A beta is generated in the ER, Golgi, and endosomal/lysosomal system. A central question involving the role of A beta in AD concerns how A beta causes disease and whether it is extracellular A beta deposition and/or intracellular A beta accumulation that initiates the disease process. The most prevalent view is that SPs are composed of extracellular deposits of secreted A beta and that A beta causes toxicity to surrounding neurons as extracellular SP. The recent emphasis on the intracellular biology of APP and A beta has led some investigators to consider the possibility that intraneuronal A beta may directly cause toxicity. In this review we will outline current knowledge of the localization of both intracellular and extracellular A beta.     

Interaction between S100A8/A9 and annexin A6 is involved in the calcium-induced cell surface exposition of S100A8/A9

The calcium binding S100A8/A9 complex (MRP8/14; calgranulin) is considered as an important proinflammatory mediator in acute and chronic inflammation and has recently gained attention as a molecular marker up-regulated in various human cancers. Here, we report that S100A8/A9 is expressed in breast cancer cell lines and is up-regulated by interleukin-1beta and tumor necrosis factor-alpha in SKBR3 and MCF-7 cells. We identified the phospholipid-binding protein annexin A6 as a potential S100A8/A9 binding protein by affinity chromatography. This finding was verified by Southwestern overlay experiments and by coimmunoprecipitation with the S100A8/A9-specific monoclonal antibody 27E10. Immunocytochemical experiments demonstrated that S100A8/A9 and annexin A6 colocalize in SKBR3 breast cancer cells predominantly in membranous structures. Upon calcium influx both S100A8/A9 and annexin A6 are exposed on the cell surface of SKBR3 cells. Subcellular fractionation studies suggested that after A23187 stimulation membrane association of S100A8/A9 is not enhanced. However, both S100A8/A9 and annexin A6 are exposed on the cell surface of SKBR3 cells upon calcium influx. Experiments with artificial liposomes indicated that S100A8/A9 is able to associate with membranes independently of both annexin A6 and independently of calcium. Finally, cell surface expression of S100A8/A9 could not be observed in A23187-treated A431 and HaCaT cells. Both cell lines are known to be devoid of annexin A6. Repression of annexin A6 expression by small interfering RNA in SKBR3 cells abolishes the cell surface exposition of S100A8/A9 upon calcium influx, suggesting that annexin A6 contributes to the calcium-dependent cell surface exposition of the membrane associated-S100A8/A9 complex.     

Two conformational states in the crystal structure of the Homo sapiens cytoplasmic ribosomal decoding A site

The decoding A site of the small ribosomal subunit is an RNA molecular switch, which monitors codon–anticodon interactions to guarantee translation fidelity. We have solved the crystal structure of an RNA fragment containing two Homo sapiens cytoplasmic A sites. Each of the two A sites presents a different conformational state. In one state, adenines A1492 and A1493 are fully bulged-out with C1409 forming a wobble-like pair to A1491. In the second state, adenines A1492 and A1493 form non-Watson–Crick pairs with C1409 and G1408, respectively while A1491 bulges out. The first state of the eukaryotic A site is, thus, basically the same as in the bacterial A site with bulging A1492 and A1493. It is the state used for recognition of the codon/anticodon complex. On the contrary, the second state of the H.sapiens cytoplasmic A site is drastically different from any of those observed for the bacterial A site without bulging A1492 and A1493.     

Injury in relation to cell organization

When a part of a Nitella cell, A, is covered with water and the rest of the cell, B, is in contact with a toxic solution there is an escape of solutes at B. This is followed by the escape of solutes at A which causes the death of A. Water enters at A, flows along inside the cell, and escapes at B carrying solutes with it. When this is prevented by covering A with mineral oil the escape of solutes at A is delayed and the life of A is correspondingly prolonged. It is remarkable that this occurs in spite of the fact that the hydrostatic pressure inside the cell (turgor) drops from 6.4 atmospheres to zero. It would seem that A might not be affected by the death of B if the escape of solutes could be prevented.