大理翠雀中的新二萜生物碱

从大理翠雀根中分到五个二萜生物碱成分,其中两个成分分别鉴定为methylly-caconitine(3)和delsemine(4)。另三个成分为新二萜生物碱,命名为大理翠雀碱甲(talitine A)、大理翠雀碱乙(talitine B)及大理翠雀碱丙(talitine C)。经质谱、红外光谱、核磁共振氢谱、核磁共振碳谱等解析,碱乙和碱丙的化学结构分别为(1)和(2),碱甲的结构另文报告。     

大花翠雀的新二萜生物碱

从大花翠雀(Delphinium grandiflorum L.)的地上部分分离到两个微量的新二萜生物碱——大花翠雀辛(grandifloricine,Ⅰ)和大花翠雀亭(grandifloritine,Ⅱ)。通过分析光谱信息测定了它们的化学结构,并通过化学反应转变为 takaosamine 而得到确证。     

秦岭翠雀花中生物碱成分的研究

从秦岭翠雀花(Delphinium giraldii Diels)全草中分得14个已知生物碱：tongolinine(1)、氨茴酰牛扁碱(anthranoyllycoctonine)(2)、牛扁碱(1ycoctonine)(3)、dihydrogadesine(4)、tatsiensine(5)、siwanine A(6)、delsemine A(7)、delsemine B(8)、甲基牛扁碱(methyllycaconitine)(9)、ajacine(10)、delajacine(11)、delsoline(12)、滇乌碱(yunaconitine)(13)和查斯曼宁(chasmanine)(14)。应用光谱和与已知品对照TLC法鉴定了所有化合物的结构。     

新疆翠雀花属新植物

本文描述了新疆翠雀花属七个新分类群,即：叶城翠雀花、假深蓝翠雀花、无腺翠雀花、赛塔城翠雀花、高翠雀花、秀丽翠雀花、白花萨乌尔翠雀花。     

Delphinium nortonii Dunn的重新描述

通过检查藏南翠雀花Delphinium nortonii 的主模式标本,发现《中国植物志》、《西藏植物志》和《Flora of China》中对该种的鉴定和描述有误。该种的花的退化雄蕊黑褐色,属于翠雀亚属的密花翠雀花组subgen. Delphinastrum sect. Elatopsis, 而上述几种植物志中长1以来将其置于退化雄蕊为蓝色或蓝紫色的翠雀组sect. Delphinastrum中。这几种植物志中记载的D. nortonii 实际上代表一个新种,即李恒翠雀花D. lihengianum。     

中国毛茛科翠雀属的分类学研究(三):拟澜沧翠雀花、磨顶山翠雀花和粗距蓝翠雀花的名实订正(英文)

通过标本室和野外观察,发现根据云南西北部德钦标本描述的拟澜沧翠雀花(Delphinium pseudothibeticum W.T.WangM.J.Warnock)与同样根据德钦标本描述的短角萼翠雀花(D.ceratophorum Franch.var.brevicorniculatum W.T.Wang)属于同一植物,故将前者处理为后者的异名;而根据德钦标本描述的磨顶山("磨顶"应为德钦县羊拉乡茂顶,故"磨顶山"实应为"茂顶山")翠雀花(D.motingshanicum W.T.WangM.J.Warnock)和粗距蓝翠雀花(D.caeruleum Jacq.ex Camb.var.crassicalcaratum W.T.WangM.J.Warnock)均与云南西北部颇为常见的宽距翠雀花(D.beesianum W.W.Smith)属于同一植物(磨顶山翠雀花为宽距翠雀花的高大个体),故将磨顶山翠雀花和粗距蓝翠雀花均处理为宽距翠雀花的异名。     

中国毛茛科翠雀属的分类学研究(十):须弥翠雀花二新异名

标本室和野外观察表明匙苞翠雀花(Delphinium subspathulatum W. T. Wang)和吉隆翠雀花(D. tabatae Tamura)与须弥翠雀花(D. himalayae Munz)属于同一种植物,故将前二者均处理为须弥翠雀花的异名。     

翠雀属一新变型

本文发表了翠雀属一新变型,即白花翠雀ＤｅｌｐｈｉｎｉｕｍｇｒａｎｄｉｆｌｏｒｍＬ．ｆ．ａｌｂｕｍＴ．Ｊ．ＦｅｎｇｅｔＪ．Ｘ．Ｈｕａｎｇ．     

拟螺距翠雀花中的新二萜生物碱

从拟螺距翠雀花的根中分离、鉴定了六个二萜生物碱成分,其中三个为已知成分,分别为methyllycaconitine(1)、ajacine(2)和delsemine(3);另三个为新二萜生物碱,分别命名为螺翠碱甲(bulleyanitine A)、螺翠碱乙(bulleyanitine B)和螺翠碱丙(bulleyanitine C),纤MS、IR、~1H NMR、~(13)C NMR及DEPT等光谱的解析证明其化学结构分别为(4)、(5)、(6)。     

中国毛茛科翠雀属的分类学研究(六):拟川西翠雀花二新异名

通过标本室和野外观察, 发现根据四川丹巴标本描述的毛茛科光果拟螺距翠雀花(Delphinium bulleyanum Forrest ex Dielsvar. leiogynum W. T. Wang)和根据四川汶川标本描述的汶川翠雀花(D. wenchuanense W. T. Wang)与此前发现分布于四川宝兴、都江堰、汶川一带的拟川西翠雀花(D. pseudotongolense W. T. Wang)没有区别, 故将二者均处理为拟川西翠雀花的异名。     

Characterization of a novel acidic protein of 38 kDa, A0, in yeast ribosomes which immunologically cross-reacts with the 13 kDa acidic ribosomal proteins, A1/A2

A new ribosomal protein of 38 kDa, named A0, was detected in yeast ribosomes on immunoblotting. The antibody used here was that against A1/A2, 13 kDa acidic ribosomal proteins which cross-reacted with A0. Although A0 and A1/A2 share common antigenic determinants, they differ in the following biochemical properties. While A1/A2 could be extracted from ribosomes with ethanol and ammonium sulfate, A0 could not. A0 gave two protein spots in a less acidic region than for A1/A2 on two-dimensional gel electrophoresis. The heterogeneity observed for A0 was ascribable to phosphorylation because one spot disappeared after treatment of the ribosomes with phosphatase. The syntheses of A0 and A1/A2 are directed by different mRNA species, as judged with a cell-free translation system, ruling out the possibility that A0 is a precursor of A1/A2. Although a mammalian ribosomal protein equivalent to A0 has been shown to be associated with 13 kDa acidic proteins in the cytoplasm, essentially no A0 was detected on immunoblotting in the yeast cytosol, while a small but detectable amount of A1/A2 was present. The possibility that A0 is a eukaryotic equivalent of L10 of Escherichia coli is discussed.     

Expression of cytochrome P450 3A in amphibian, rat, and human kidney.

Family 3A mammalian liver cytochromes P450 (3A1, rat; 3A3/4, human) catalyze the 6 beta-hydroxylation of endogenous steroids and are steroid inducible. Our recent finding that A6 cells (a toad kidney epithelial cell line) contain corticosterone 6 beta-hydroxylase activity as a steroid-inducible microsomal cytochrome P450 raised the possibility that corticosterone 6 beta-hydroxylase activity in the A6 cells is catalyzed by a member of the 3A family. We found that incubation of A6 cell microsomes from dexamethasone-induced cells with antibodies against family 3A proteins specifically inhibited corticosterone 6 beta-hydroxylase activity. Microsomes from A6 cells analyzed on immunoblots developed with family 3A specific antibodies revealed immunoreactive proteins and treatment of A6 with corticosterone or dexamethasone increased the amounts of 3A immunoreactive protein(s). Furthermore, A6 RNA hybridized with 3A cDNAs on Northern blots and genomic DNA from A6 cells hybridized with a 3A cDNA on a Southern blot. Thus, toad kidney A6 cells express a family 3A P450 that is immunochemically, functionally, and genetically related to the mammalian liver 3A proteins. Prompted by these findings in amphibian kidney, we examined mammalian kidney for evidence of family 3A proteins. Immunocytochemical studies of frozen cryostat sections of normal adult rat kidney incubated with 3A1 antibody showed immunoreactivity only with collecting duct. Immunoblot analysis of human kidney microsomes found three protein bands representing 3A3/4, 3A5, and a 53-kDa Mr protein immunoreactive with human 3A antibody. An unexpected finding was the polymorphic expression of 3A3/4 in human kidney with only one of seven (14%) adult human kidneys tested expressing this protein while 3A5, a protein which is polymorphically expressed in adult human livers, was routinely present in the adult human kidney samples tested. Since human fetal liver contains a family 3A P450 we examined human fetal kidney microsomes by immunoblot analysis with human liver 3A antibody and found expression of a protein tentatively identified as 3A7. Thus, like A6 amphibian cells, family 3A P450 proteins and mRNAs are prominent, functional components in the kidney of mammals, including man.     

Glycosylated minor components of human adult hemoglobin. Purification, identification, and partial structural analysis

Human hemolysate contains several minor components designated Hb A1a, Hb A1b, Hb A1c, which are post-translational modifications of the major hemoglobin component A0. Individuals with diabetes mellitus have elevated levels of Hb A1c, a hemoglobin modified with a glucose moiety at the NH2 terminus of each beta chain. A new chromatographic technique using Bio-Rex 70 is described which not only allows complete separation of Hb A1a from Hb A1b but also resolution of Hb A1a into two components, designated Hb A1a1 and Hb A1a2. Carbohydrate determinations with the thiobarbituric acid procedure revealed that Hb A1a1, Hb A1a2, and Hb A1b as well as Hb A1c were glycosylated. Total phosphate analysis revealed 2.06 and 1.01 mol of phosphorus/alphabeta dimer for Hb A1a1 and Hb A1a2 respectively; Hb A1b and Hb A1c contained no detectable phosphate. Hemoglobin incubated with D-[14C]glucose-6-P co-chromatographs precisely with Hb A1a2, strongly suggesting that Hb A1a2 is glucose-6-P hemoglobin. Levels of Hb A1a1 and Hb A1a2 are normal in individuals with diabetes mellitus. Furthermore, diabetic red cells contain normal levels of glucose-6-P. Therefore, glucose-6-P hemoglobin does not serve as a significant precursor to Hb A1c. Instead Hb A1c is formed by the direct reaction of hemoglobin with glucose. This suggests that hemoglobin can serve as a model system for nonenzymatic glycosylation of protein.     

Altered interactions between the A1 and A2 subunits of factor VIIIa following cleavage of A1 subunit by factor Xa

Factor VIIIa consists of subunits designated A1, A2, and A3-C1-C2. The limited cofactor activity observed with the isolated A2 subunit is markedly enhanced by the A1 subunit. A truncated A1 (A1(336)) was previously shown to possess similar affinity for A2 and retain approximately 60% of its A2 stimulatory activity. We now identify a second site in A1 at Lys(36) that is cleaved by factor Xa. A1 truncated at both cleavage sites (A1(37-336)) showed little if any affinity for A2 (K(d)>2 microm), whereas factor VIIIa reconstituted with A2 plus A1(37-336)/A3-C1-C2 dimer demonstrated significant cofactor activity ( approximately 30% that of factor VIIIa reconstituted with native A1) in a factor Xa generation assay. These affinity values were consistent with values obtained by fluorescence energy transfer using acrylodan-labeled A2 and fluorescein-labeled A1. In contrast, factor VIIIa reconstituted with A1(37-336) showed little activity in a one-stage clotting assay. This resulted in part from a 5-fold increase in K(m) for factor X when A1 was cleaved at Arg(336). These findings suggest that both A1 termini are necessary for functional interaction of A1 with A2. Furthermore, the C terminus of A1 contributes to the K(m) for factor X binding to factor Xase, and this parameter is critical for activity assessed in plasma-based assays.     

Vaccinia Virus Virion Membrane Biogenesis Protein A11 Associates with Viral Membranes in a Manner That Requires the Expression of Another Membrane Biogenesis Protein,A6

A group of vaccinia virus (VACV) proteins, including A11, L2, and A6, are required for biogenesis of the primary envelope of VACV, specifically, for the acquisition of viral membrane precursors. However, the interconnection among these proteins is unknown and, with the exception of L2, the connection of these proteins with membranes is also unknown. In this study, prompted by the findings that A6 coprecipitated A11 and that the cellular distribution of A11 was dramatically altered by repression of A6 expression, we studied the localization of A11 in cells by using immunofluorescence and cell fractionation analysis. A11 was found to associate with membranes and colocalize with virion membrane proteins in viral replication factories during normal VACV replication. A11 partitioned almost equally between the detergent and aqueous phases upon Triton X-114 phase separation, demonstrating an intrinsic affinity with lipids. However, in the absence of infection or VACV late protein synthesis, A11 did not associate with cellular membranes. Furthermore, when A6 expression was repressed, A11 did not colocalize with any viral membrane proteins or associate with membranes. In contrast, when virion envelope formation was blocked at a later step by repression of A14 expression or by rifampin treatment, A11 colocalized with virion membrane proteins in the factories. Altogether, our data showed that A11 associates with viral membranes during VACV replication, and this association requires A6 expression. This study provides a physical connection between A11 and viral membranes and suggests that A6 regulates A11 membrane association.     

Vaccinia virus A26 and A27 proteins form a stable complex tethered to mature virions by association with the A17 transmembrane protein

During vaccinia virus replication, mature virions (MVs) are wrapped with cellular membranes, transported to the periphery, and exported as extracellular virions (EVs) that mediate spread. The A26 protein is unusual in that it is present in MVs but not EVs. This distribution led to a proposal that A26 negatively regulates wrapping. A26 also has roles in the attachment of MVs to the cell surface and incorporation of MVs into proteinaceous A-type inclusions in some orthopoxvirus species. However, A26 lacks a transmembrane domain, and nothing is known regarding how it associates with the MV, regulates incorporation of the MV into inclusions, and possibly prevents EV formation. Here, we provide evidence that A26 forms a disulfide-bonded complex with A27 that is anchored to the MV through a noncovalent interaction with the A17 transmembrane protein. In the absence of A27, A26 was unstable, and only small amounts were detected. The interaction of A26 with A27 depended on a C-terminal segment of A26 with 45% amino acid identity to A27. Deletion of A26 failed to enhance EV formation by vaccinia virus, as had been predicted. Nevertheless, the interaction of A26 and A27 may have functional significance, since each is thought to mediate binding to cells through interaction with laminin and heparan sulfate, respectively. We also found that A26 formed a noncovalent complex with A25, a truncated form of the cowpox virus A-type inclusion matrix protein. The latter association suggests a mechanism for incorporation of virions into A-type inclusions in other orthopoxvirus strains.     

Cell-mediated Immunity in Patients Positive for Hepatitis-associated Antigen

Cell-mediated immunity to antigens prepared from both serum and liver of patients positive for hepatitis-associated antigen (H.A.A.) was measured by using the leucocyte migration test. Altogether, 43 patients with H.A.A.-positive acute and chronic liver disease, eight with serum antibody to H.A.A., and 13 controls were studied. The cell-mediated immunity detected was specific for H.A.A. or other antigenic determinants of the associated infective agent and could be found only in patients with evidence of previous contact with H.A.A.Cell-mediated immunity to the H.A.A.-positive test antigens was found in all but one of the patients with acute hepatitis, in about half of the patients with chronic aggressive hepatitis or cirrhosis, rarely in those with chronic persistent hepatitis, and in none of the apparently healthy carriers.Our results support the hypothesis that the cellular immune response plays an important part in the clearance of the infective agent from H.A.A.-positive patients and in the pathogenesis of the associated liver cell injury.     

Interaction of human pancreatic ribonuclease with human ribonuclease inhibitor. Generation of inhibitor-resistant cytotoxic variants

Mammalian ribonucleases interact very strongly with the intracellular ribonuclease inhibitor (RI). Eukaryotic cells exposed to mammalian ribonucleases are protected from their cytotoxic action by the intracellular inhibition of ribonucleases by RI. Human pancreatic ribonuclease (HPR) is structurally and functionally very similar to bovine RNase A and interacts with human RI with a high affinity. In the current study, we have investigated the involvement of Lys-7, Gln-11, Asn-71, Asn-88, Gly-89, Ser-90, and Glu-111 in HPR in its interaction with human ribonuclease inhibitor. These contact residues were mutated either individually or in combination to generate mutants K7A, Q11A, N71A, E111A, N88R, G89R, S90R, K7A/E111A, Q11A/E111A, N71A/E111A, K7A/N71A/E111A, Q11A/N71A/E111A, and K7A/Q11A/N71A/E111A. Out of these, eight mutants, K7A, Q11A, N71A, S90R, E111A, Q11A/E111A, N71A/E111A, and K7A/N71A/E111A, showed an ability to evade RI more than the wild type HPR, with the triple mutant K7A/N71A/E111A having the maximum RI resistance. As a result, these variants exhibited higher cytotoxic activity than wild type HPR. The mutation of Gly-89 in HPR produced no change in the sensitivity of HPR for RI, whereas it has been reported that mutating the equivalent residue Gly-88 in RNase A yielded a variant with increased RI resistance and cytotoxicity. Hence, despite its considerable homology with RNase A, HPR shows differences in its interaction with RI. We demonstrate that interaction between human pancreatic ribonuclease and RI can be disrupted by mutating residues that are involved in HPR-RI binding. The inhibitor-resistant cytotoxic HPR mutants should be useful in developing therapeutic molecules.