Antiviral resistance by the polyinosinic acid-poly (1-vinylcytosine) complex

The antiviral activities of analogs of the double-stranded complex of polyinosinic and polycytidylic acids [poly(I).poly(C)], which is a potent interferon inducer, have been studied. Structural changes that modify the polymer backbone substantially, such as loops or 2' --> 5' phosphodiester bonds, lead to decreased antiviral activity. Unexpectedly, however, the complex of polyinosinic acid and poly(1-vinylcytosine), which is only a much more distantly related analog of poly(I) . poly(C), shows high activity. It is postulated that the high activity is related to the reduction of the charge/mass ratio and to the existence of this complex in an aggregated state; these are two factors that generally enhance the uptake of compo unds by cells.     

Tolerance to polyinosinic-polycytidylic acid in NZB-NZW mice

Immunological tolerance to polyinosinic * polycytidylic acid was induced in NZB/ NZW mice. This is the first experimental induction of tolerance to a nucleic antigen.     

Coding channels in the taste system of the rat

Basic taste qualities are thought to be perceived independently, yet discrete neural coding channels have not been demonstrated in the central nervous system. The response profiles of taste cells in the nucleus tractus solitarius (NTS) of the rat were categorized into four groups, and the effects of amiloride, a passive sodium channel blocker, on each were determined. NTS neurons that responded specifically to sodium chloride (NaCl) or to NaCl and sugars were suppressed by amiloride; those broadly sensitive to salts, acids, and bitter stimuli were unaffected. Moreover, the response profile evoked by NaCl lost its distinctiveness after treatment with amiloride, becoming similar to those evoked by acids and quinine. Receptors that respond to sodium must relay their information through independent coding channels to identifiable subgroups of NTS neurons, the activity of which is responsible for the perception of saltiness.     

Polyinosinic acid--polycytidylic acid: inhibition of DNA synthesis stimulated by isoproterenol

Polyinosinic acid * polycytidylic acid (poly I * poly C) inhibits isoproterenol-stimulated DNA synthesis in salivary glands of mice. A single intraperitoneal injection of 250 micrograms of poly I * poly C inhibits the stimulation of DNA synthesis when given 10 minutes before isoproterenol or at any time during the 20-hour lag period between the injection of isoproterenol and the subsequent DNA synthesis. The inhibition caused by poly I * poly C is not due to a generalized toxic action but seems to be exerted through a relatively selective mechanism. Polyuridylic acid and diethylaminoethyl dextran are less effective in inhibiting isoproterenol-stimulated DNA synthesis.     

Amino acid homology between the encephalitogenic site of myelin basic protein and virus: mechanism for autoimmunity

Amino acid sequence homology was found between viral and host encephalitogenic protein. Immune responses were then generated in rabbits by using the viral peptide that cross-reacts with the self protein. Mononuclear cell infiltration was observed in the central nervous systems of animals immunized with the viral peptide. Myelin basic protein (MBP) is a host protein whose encephalitogenic site of ten amino acids induces experimental allergic encephalomyelitis. By computer analysis, hepatitis B virus polymerase (HBVP) was found to share six consecutive amino acids with the encephalitogenic site of rabbit MBP. Rabbits given injections of a selected eight- or ten-amino acid peptide from HBVP made antibody that reacted with the predetermined sequences of HBVP and also with native MBP. Peripheral blood mononuclear cells from the immunized rabbits proliferated when incubated with either MBP or HBVP. Central nervous system tissue taken from these rabbits had a histologic picture reminiscent of experimental allergic encephalomyelitis. Thus, viral infection may trigger the production of antibodies and mononuclear cells that cross-react with self proteins by a mechanism termed molecular mimicry. Tissue injury from the resultant autoallergic event can take place in the absence of the infectious virus that initiated the immune response.     

Host proteasomal degradation generates amino acids essential for intracellular bacterial growth

Legionella pneumophila proliferates in environmental amoeba and human cells within the Legionella-containing vacuole (LCV). The exported AnkB F-box effector of L. pneumophila is anchored into the LCV membrane by host-mediated farnesylation. Here, we report that host proteasomal degradation of Lys(48)-linked polyubiquitinated proteins, assembled on the LCV by AnkB, generates amino acids required for intracellular bacterial proliferation. The severe defect of the ankB null mutant in proliferation within amoeba and human cells is rescued by supplementation of a mixture of amino acids or cysteine, serine, pyruvate, or citrate, similar to rescue by genetic complementation. Defect of the ankB mutant in intrapulmonary proliferation in mice is rescued upon injection of a mixture of amino acids or cysteine. Therefore, Legionella promotes eukaryotic proteasomal degradation to generate amino acids needed as carbon and energy sources for bacterial proliferation within evolutionarily distant hosts.     

Template-directed oligomerization catalyzed by a polynucleotide analog

A pyrophosphate-linked analog of polycytidylic acid has been synthesized and shown to catalyze the oligomerization of the complementary monomer 2'-deoxyguanosine 3',5'-bisphosphoimidazolide. Analogs of polynucleotides are of interest in studies of the origins of life as possible precursors of the first RNA molecules. These results demonstrate that such molecules are capable of serving as templates for further synthesis.     

Expanding the genetic code of Escherichia coli

A unique transfer RNA (tRNA)/aminoacyl-tRNA synthetase pair has been generated that expands the number of genetically encoded amino acids in Escherichia coli. When introduced into E. coli, this pair leads to the in vivo incorporation of the synthetic amino acid O-methyl-l-tyrosine into protein in response to an amber nonsense codon. The fidelity of translation is greater than 99%, as determined by analysis of dihydrofolate reductase containing the unnatural amino acid. This approach should provide a general method for increasing the genetic repertoire of living cells to include a variety of amino acids with novel structural, chemical, and physical properties not found in the common 20 amino acids.     

The Rag GTPases bind raptor and mediate amino acid signaling to mTORC1

The multiprotein mTORC1 protein kinase complex is the central component of a pathway that promotes growth in response to insulin, energy levels, and amino acids and is deregulated in common cancers. We find that the Rag proteins--a family of four related small guanosine triphosphatases (GTPases)--interact with mTORC1 in an amino acid-sensitive manner and are necessary for the activation of the mTORC1 pathway by amino acids. A Rag mutant that is constitutively bound to guanosine triphosphate interacted strongly with mTORC1, and its expression within cells made the mTORC1 pathway resistant to amino acid deprivation. Conversely, expression of a guanosine diphosphate-bound Rag mutant prevented stimulation of mTORC1 by amino acids. The Rag proteins do not directly stimulate the kinase activity of mTORC1, but, like amino acids, promote the intracellular localization of mTOR to a compartment that also contains its activator Rheb.     

水稻氨基酸透性酶基因OsAAP14启动子克隆及时空表达特性分析

【目的】克隆水稻氨基酸透性酶基因OsAAP14的启动子序列并分析其时空表达特性，为解析氨基酸转运基因生物学功能及了解水稻对有机氮源的响应和氨基酸吸收利用提供理论依据。【方法】PCR扩增OsAAP14基因的启动子序列，并与pCAMBIA1391Z空载体质粒连接构建出启动子-GUS表达载体，并通过农杆菌介导侵染水稻中花11愈伤组织，获得OsAAP14基因的启动子-GUS转基因植株，通过对转基因植株不同组织部位进行GUS染色，并采用石蜡切片技术观察各组织细胞部位表达，经5种不同氨基酸处理后进行根部GUS染色，结合实时荧光定量PCR检测OsAAP14基因在GUS染色部位的相对表达量，最后综合分析该基因的时空表达特性。【结果】克隆获得OsAAP14基因启动子序列为1993 bp，与参考序列日本晴OsAAP14（LOC_Os04g56470）序列一致。该启动子序列含有MBS、P-box、ABRE、CGTCA-motif等激素或胁迫响应元件。从OsAAP14基因的启动子-GUS植株T₀代中鉴定获得16个阳性转基因株系。T₁代材料不同组织部位GUS染色及实时荧光定量PCR结果均显示，OsAAP14基因在水稻芽伸长的基部和叶片相对表达量较高，在根、叶鞘和穗也有一定表达，但在茎中的相对表达量最低。石蜡切片分析结果显示，OsAAP14基因在根部皮层薄壁细胞、叶片的叶肉细胞、穗的颖壳内部细胞有较高的表达。碱性氨基酸的组氨酸处理下根中的OsAAP14基因表达量随着处理时间的增加显著提高（P<0.05，下同），且赤霉素和脱落酸处理下，根中的OsAAP14基因相对表达量也显著提高。【结论】OsAAP14基因在水稻不同组织均有表达，正常情况（未处理）下在水稻基部和叶片中相对表达量较高，但外源氨基酸和激素处理时，在根中该基因被诱导表达上调，说明OsAAP14基因正常情况下可能主要参与调控水稻地上部分氨基酸的运输，但当外界氨基酸和激素含量增加时则参与调控水稻根部氨基酸的运输。     

Arachidonic acid and other fatty acids directly activate potassium channels in smooth muscle cells

Arachidonic acid, as well as fatty acids that are not substrates for cyclooxygenase and lipoxygenase enzymes, activated a specific type of potassium channel in freshly dissociated smooth muscle cells. Activation occurred in excised membrane patches in the absence of calcium and all nucleotides. Therefore signal transduction pathways that require such soluble factors, including the NADPH-dependent cytochrome P450 pathway, do not mediate the response. Thus, fatty acids directly activate potassium channels and so may constitute a class of signal molecules that regulate ion channels.     

开放式空气中CO2浓度增高（FACE）对常规粳稻蛋白质和氨基酸含量的影响

为了明确未来大气CO2浓度升高对水稻蛋白质营养品质的影响,2009年利用稻田开放式空气CO2浓度增高(FACE,FreeAirCO2 Enrichment)系统,以武运粳21、扬辐粳8号和武粳15为供试品种,研究大田生长期CO2浓度升高200μmol.mol-1对常规粳稻蛋白质营养品质的影响。结果表明:大气CO2浓度增加使所有供试品种精米蛋白质含量平均下降5.6%,使氨基酸、必需和非必需氨基酸总量平均分别下降7.6%、6.7%和7.9%,均达极显著水平。大气CO2浓度增加使供试品种精米必需氨基酸占氨基酸总量百分比显著增加,使非必需氨基酸占氨基酸总量百分比显著下降,但对精米中必需和非必需氨基酸的相对含量无显著影响。从氨基酸组分看,大气CO2浓度增加使供试品种精米中7种必需氨基酸和8种非必需氨基酸的含量均显著或极显著下降。CO2处理与品种对精米蛋白质含量、氨基酸总量、必需和非必需氨基酸总量以及部分氨基酸组分有一定的互作效应,武运粳21上述参数对高浓度CO2的响应大于扬辐粳8号或武粳15对应参数的响应。以上结果说明本世纪中叶大气中的CO2浓度将使粳稻蛋白质营养品质下降,不同品种下降幅度存在一定差异。     

Efficient metal-ion catalyzed template-directed oligonucleotide synthesis

The Pb2+ and Zn2+ ions are efficient catalysts for the polycytidylic acid-directed polymerization of an activated guanylic acid derivative, guanosine 5'-phosphorimidazolide. The products include oligomers of 30 to 40 units in length. The nucleotide residues are predominantly 2'-5' linked when Pb2+ is the catalyst, and predominantly 3'-5' linked in the presence of Zn2+. The significance of these results in the context of the prebiotic evolution of RNA polymerase is discussed.     

植物中游离氨基酸在水淹胁迫下响应的研究进展

游离氨基酸含量的变化可以用来指示环境压力的变化。近年来,研究植物形态、分布格局等对水淹的响应较多,但对植物中氨基酸代谢机制的研究较少。综述水淹下植物中氨基酸代谢的变化,主要包括植物组织中游离氨基酸GABA、Glu在水淹胁迫下的响应,以及植物中其他游离氨基酸的响应情况,提出以后研究应关注复合压力胁迫下植物中游离氨基酸的响应。     

Absence of glycerol teichoic acids in certain oral streptococci

Glycerol teichoic acids were not detected immunochemically or chemically in phenol-water, hot saline (Rantz and Randall), or supernatant fluids of disrupted cells of Streptococcus mitis. Thus teichoic acids do not appear to be found in most Gram-positive bacteria, as has been suggested.     

Acetylcholine responses in L cells

L cells, a family of continuous cell lines of mouse fibroblastic origin, generate a prolonged active membrane hyperpolarization (the hyperpolarizing activation response) when stimulated mechanically or electrically. lontophoretically applied acetylcholine elicits a similar response; atropine blocks the acetylcholine but not the electrically or mechanically elicited responses. The hyperpolarizing activation response can also be elicited by electrical, mechanical, or acetylcholine stimulation of cells adjacent to the recorded cell. Propagation of the response from one cell to another is not dependent on direct electrical coupling between cells and is not blocked by application of a bath containing atropine or curare. These results show that L cells are capable of generating an active electrical response. that they are sensitive to at least one neurotransmitter (acetylcholine), and that humorally mediated interaction (probably noncholinergic) between L cells occurs.     

Homeostasis of the antibody response: immunoregulation by NK cells

When injected into mice, the synthetic double-stranded polynucleotide poly(inosinic) X poly(cytidylic) acid induces high natural killer (NK) cell activity within 4 to 12 hours. Induction of NK activity in mice immunized 2 or 3 days previously, or the addition of NK cells to cultures immunized in vitro 2 or 3 days previously, promotes early termination of the ongoing primary immunoglobulin M antibody response. A target for NK cells is a population of accessory cells that has interacted with antigen and is necessary for sustaining the antibody response. The inference is strong that NK cells induced normally by immunization also terminate the usual antibody response in vivo by elimination of antigen-exposed accessory cells.     

血液和组织标本中脂肪酸组成的毛细管柱气相色谱分析

用氯仿甲醇抽提、三氟化硼－乙醚甲醇酯化，然后利用毛细管柱和程序升温的方法进行气相色谱分析，建立了血浆和红细胞膜总脂以及组织磷脂和中性脂脂肪酸组成的毛细管柱气相色谱分析法。图谱表明各峰间分离效果良好、峰形尖锐、干扰少。各种脂肪酸的最小检测限均可达１０－（－１１）ｇ／ｓ，ＦＩＤ对各脂肪酸的响应均呈线性。实验重复性好，回收率达定量分析要求．     

Insulin receptor of fat cells in insulin-resistant metabolic states

A diminished response to insulin is exhibited by isolated fat cells obtained from rats that have been either starved, or treated with prednisone, or made diabetic by administration of streptozotocin. This decrease in response is not accompanied by changes in the quantity of insulin receptor of these cells or in the affinity of these receptors for insulin. Similarly, the decreased responsiveness to insulin of fat cells obtained from certain species (hamster, rabbit, mouse, guinea pig) is not explainable in terms of alterations of the insulin receptor.     

The intervening sequence RNA of Tetrahymena is an enzyme

A shortened form of the self-splicing ribosomal RNA (rRNA) intervening sequence of Tetrahymena thermophila acts as an enzyme in vitro. The enzyme catalyzes the cleavage and rejoining of oligonucleotide substrates in a sequence-dependent manner with Km = 42 microM and kcat = 2 min-1. The reaction mechanism resembles that of rRNA precursor self-splicing. With pentacytidylic acid as the substrate, successive cleavage and rejoining reactions lead to the synthesis of polycytidylic acid. Thus, the RNA molecule can act as an RNA polymerase, differing from the protein enzyme in that it uses an internal rather than an external template. At pH 9, the same RNA enzyme has activity as a sequence-specific ribonuclease.