Thrombin-stimulated cell proliferation mediated through activation of Ras/Raf/MEK/MAPK pathway in canine cultured tracheal smooth muscle cells.

The elevated level of thrombin has been detected in the airway fluids of asthmatic patients and shown to stimulate cell proliferation in tracheal smooth muscle cells (TSMCs). However, the implication of thrombin in the cell proliferation was not completely understood. In this study, thrombin stimulated [3H]thymidine incorporation and p42/p44 mitogen-activated protein kinase (MAPK) phosphorylation in a time- and concentration-dependent manner in TSMCs. Pretreatment of TSMCs with pertussis toxin (PTX) significantly inhibited [3H]thymidine incorporation and phosphorylation of MAPK induced by thrombin. These responses were attenuated by tyrosine kinase inhibitors genistein and herbimycin A, phosphatidyl inositide (PI)-phospholipase C (PLC) inhibitor U73122, protein kinase C inhibitor GF109203X, removal of Ca2+ by addition of BAPTA/AM plus EGTA, PI 3-kinase inhibitors wortmannin and LY294002, and inhibitor of MEK1/2 PD98059. Furthermore, overexpression of dominant negative mutants, H-Ras-15A and Raf-N4, significantly suppressed p42/p44 MAPK activation induced by thrombin and PDGF-BB, indicating that Ras and Raf may be required for activation of these kinases. These results conclude that the mitogenic effect of thrombin was mediated through the activation of Ras/Raf/MEK/MAPK pathway. Thrombin-mediated MAPK activation was modulated by PI-PLC, Ca2+, PKC, tyrosine kinase, and PI 3-kinase associated with cell proliferation in canine cultured TSMCs.     

Impacts of natural disturbance on fish communities in the Tachia River, Taiwan

Extensive landslides triggered by the Chi-Chi earthquake of 21 September 1999 introduced debris into the middle section of Tachia River, while debris flow was subsequently induced by Typhoon Toraji on 30 July 2001. We compared population size, species composition, and diversity of the fish community during a six-year period before and after these disturbances. The dominant taxa were Acrossocheilus paradoxus, Crossostoma lacustre, Hemimyzon formosanum, Rhinogobius brunneus, Varicorhinus barbatulus and Zacco pachycephalus. H. formosanum and R. brunneus increased then decreased suddenly in abundance following the earthquake and the typhoon. Our results suggest that both resistance and resilience were important in maintaining long-term fish community structure. Fish community resistance at station 1 (downstream of the disturbance) was lower than at station 2 and station 3 (at and upstream of the disturbance). Fish communities recovered quickly after a few months, possibly reflecting a correlation between assemblage composition and seasonal variation. Our study illustrates the ecological variability that can be induced by hydrologic and evolutionary processes in a stream. Relative positions of habitats provide a spatial framework for evaluating stress effects of abiotic and biotic factors in regulating population size and community succession patterns.     

Genetic control of T and B lymphocyte activation in nonobese diabetic mice.

Type 1 diabetes in nonobese diabetic (NOD) mice is characterized by the infiltration of T and B cells into pancreatic islets. T cells bearing the TCR Vbeta3 chain are disproportionately represented in the earliest stages of islet infiltration (insulitis) despite clonal deletion of most Vbeta3(+) immature thymocytes by the mammary tumor virus-3 (Mtv-3) superantigen (SAg). In this report we showed that a high frequency of NOD Vbeta3(+) T cells that escape deletion are activated in vivo and that this phenotype is linked to the Mtv-3 locus. One potential mechanism of SAg presentation to peripheral T cells is by activated B cells. Consistent with this idea, we found that NOD mice harbor a significantly higher frequency of activated B cells than nondiabetes-prone strains. These activated NOD B cells expressed cell surface molecules consistent with APC function. At the molecular level, the IgH repertoire of activated B cells in NOD mice was equivalent to resting B cells, suggesting a polyclonal response in vivo. Genetic analysis of the activated B cell phenotype showed linkage to Idd1, the NOD MHC haplotype (H-2(g7)). Finally, Vbeta3(+) thymocyte deletion and peripheral T cell activation did not require B cells, suggesting that other APC populations are sufficient to generate both Mtv-3-linked phenotypes. These data provide insight into the genetic regulation of NOD autoreactive lymphocyte activation that may contribute to failure of peripheral tolerance and the pathogenesis of type I diabetes.     

Isolation of an Insertion Sequence from Ralstonia solanacearum Race 1 and Its Potential Use for Strain Characterization and Detection

A new insertion sequence (IS), IS1405, was isolated and characterized from a Ralstonia solanacearum race 1 strain by the method of insertional inactivation of the sacB gene. Sequence analysis indicated that the IS is closely related to the members of IS5 family, but the extent of nucleotide sequence identity in 5′ and 3′ noncoding regions between IS1405 and other members of IS5 family is only 23 to 31%. Nucleotide sequences of these regions were used to design specific oligonucleotide primers for detection of race 1 strains by PCR. The PCR amplified a specific DNA fragment for all R. solanacearum race 1 strains tested, and no amplification was observed with some other plant-pathogenic bacteria. Analysis of nucleotide sequences flanking IS1405 and additional five endogenous IS1405s that reside in the chromosome of R. solanacearum race 1 strains indicated that IS1405 prefers a target site of CTAR and has two different insertional orientations with respect to this target site. Restriction fragment length polymorphism (RFLP) pattern analysis using IS1405 as a probe revealed extensive genetic variation among strains of R. solanacearum race 1 isolated from eight different host plants in Taiwan. The RFLP patterns were then used to subdivide the race 1 strains into two groups and several subgroups, which allowed for tracking different subgroup strains of R. solanacearum through a host plant community. Furthermore, specific insertion sites of IS1405 in certain subgroups were used as a genetic marker to develop subgroup-specific primers for detection of R. solanacearum, and thus, the subgroup strains can be easily identified through a rapid PCR assay rather than RFLP analysis.     

香港大榄郊野公园的植物组成与植被数量分类

通过对香港大榄郊野公园森林群落的样地调查, 利用组平均法和除趋势对应分析两种方法, 对样方进行聚类与排序分析。把35个样地划分为11个植被类型, 森林群落可划分为台湾相思(Acacia confusa Merr.)林、红胶木(Lophostemon confertus(R. Br.) Peter G. Wils on ex J. T. Waterh.)林、木荷(Schima superba Gardner ex Champ.)林、柠檬桉(Eucalyptus citriodora Hook.)林、湿地松+鸭脚木(Pinus elliottii Engelm.+ Schefflera octophylla (Lour.) Harms .)林、土沉香(Aquilaria sinensis (Lour.) Spreng.)林、毛叶桉(Eucalyptus torel liana F. Muel l.)林; 灌木群落分为山油柑+越南叶下珠(Acronychia pedunculata (L.) Miq.+ Phyl lanthus cochinchinensis (Lour.) Spreng.)矮树灌丛、豺皮樟+桃金娘(Li tsea rotoundifol ia (Nees) Hemsl . var. ob longi folia (Nees) C. K. Allen+Rhodomyrtus tomentosa (Al ton) Hassk.)矮树灌丛、大头茶(Gordonia axillaris (Roxb. ex Ker Gawl .) D. Dietr.)矮树灌丛、桃金娘+芒萁(Rhodomyrtus tomentosa (Alton) Hassk.+Dicranopteris dichotoma (Thunb.) Bernh.)灌草丛。DCA排序结果反映海拔高度与各个森林群落的分布格局和物种组成关系不大, 而环境 湿度是影响森林群落物种组成的主要因素。多样性分析结果显示, 台湾相思林和土沉香林多样性指数最高, 说明台湾相思林逐渐向次生林演变, 除了红胶木林和少数郁闭度低的台湾相思林中有强阳生性树种外, 大部分人工林下植物以阳生性稍耐阴的树种为主。调查结果显示, 大榄郊野公园植物生长条件东部优于西部, 西部的物种丰富度和多样性指数较低, 主要以人工林和灌丛为主, 加上山火频繁发生, 导致长期处于先锋阶段。     

滇黄芩中的两个酮甙

从龙胆科滇黄芩属植物滇黄芩(Veratrilla baillonii Franch.)中分离得到两个新的(口山)酮甙,分别命名为:滇黄芩甙C(I)和滇黄芩甙D(Ⅱ).经各种光谱解析及化学反应证明,其结构推定为:2,3,4,5-四甲氧(口山)酮-1-O-β-D-吡喃葡萄糖基(6←l)-β-D-吡喃木糖甙(I)和3,4,5-三甲氧基(口山)酮-1-O-β-D-吡喃葡萄糖基(6←1)-β-D-吡喃本糖甙(Ⅱ).     

Prevalence of the virulence plasmids of nontyphoid Salmonella in the serovars isolated from humans and their association with bacteremia.

To determine if the virulence plasmid is one of the elements contributing to Salmonella bacteremia in humans, 436 clinical Salmonella isolates of different serovars were examined by a specific multiplex polymerase chain reaction assay for the presence of a virulence plasmid. These serovars showed differences in their ability to produce particular disease syndrome in humans. In the serovars usually causing bacteremia without concomitant gastroenteritis (primary bacteremia), i.e., S. choleraesuis, S. dublin, and S. enteritidis in this study, the rate of virulence plasmid carriage was 100%, while among those occasionally generating bacteremia following an episode of gastroenteritis (secondary bacteremia), the majority were plasmidless. Only a portion of S. typhimurium strains harbored a virulence plasmid; however, the rates of virulence plasmid carriage in S. typhimurium were not statistically different between non-fecal and fecal isolates (90% vs. 85%, 0.1 < P < 0.9). These results indicate that the virulence plasmids may be important for primary bacteremia, but not secondary bacteremia, to occur.     

Sodium Butyrate Induces Apoptosis in MSN Neuroblastoma Cells in a Calcium Independent Pathway

Sodium butyrate (NaBt), a histone deacetylase inhibitor, can cause apoptosis in a number of cancer cells. However, the mechanism of this action is poorly understood. Increased intracellular [Ca²⁺] level has been suggested as a likely mechanism, but there is little corroborating data. In this report we provide evidence that NaBt-treated MSN neuroblastoma cells undergo massive apoptosis in the presence of serum and regardless of external or internal [Ca²⁺] levels. Presented data suggest that apoptotic effect of NaBt is both time- and dose-dependent (LD₅₀ 1 mM); and that, presence of serum or cAMP, a second messenger molecule that modulates the apoptotic program in a wide variety of cells could not circumvent the apoptotic effect of NaBt. Our findings suggest that NaBt-induced apoptosis in MSN neuroblastoma cells occurs via a pathway that is independent of Ca²⁺flux, intracellular [Ca²⁺] or cAMP levels. Further, we also present data that exclude a role for PKC or histones acetylation.Special issue dedicated to Lawrence F. Eng