马铃薯人工种子的研究

本研究以虎头（Ｈ）、克四（Ｋ）和Ｆａｖｏｒｉｔａ（Ｆ）三个品种的马铃薯不定芽在液体培养条件下诱导生成的微型薯为试材,经筛选后获得的大小一致的微型薯经过适度的低温处理后,用４％的海藻酸钠和２％的氯化钙溶液,在附加一定的植物激素条件下,进行人。种皮包埋形成人工种子。在无菌条件下,三个品种的人工种子萌发率均达９０％以上。在土壤中,人工种子萌发率可达６０％以上,萌发的人工种子中８０％能够形成生长发育正常的完整植株。     

白细胞介素－2的PEG化

用自制的氨基ＰＥＧ化试剂ｒＩＬ－２进行化学修饰,研究了试剂浓度,溶液ｐＨ,反应时间等与ＰＥｃａ－ｒＩＬ－２产率及ＩＬ－２活性保持之间的关系,建立了一套获得稳定修饰度的ＰＥＧ－ｒＩＬ－２的方法。研究发现,反应时间跟修饰度关系不大;溶液ｐＨ对修饰度有一定的影响,中性ｐＨ以上反应都可进行;而试剂浓度直接决定修饰度的高低,过量越多,修饰度越高,而生物活性保留也越低;但低度修饰,对活性几乎没有影响,可保留活性在９５％左右。     

Multiplicity of brain cysteine sulfinic acid decarboxylase — Purification,characterization and subunit structure

Cysteine sulfinic acid decarboxylase (CSAD), the rate-limiting enzyme in taurine biosynthesis, appears to be present in the brain in multiple isoforms. Two distinct forms of CSAD, referred to as CSAD I and CSAD II, were obtained on Sephadex G-100 column. CSAD I and CSAD II differ in (1) the elution profile on Sephadex G-100 column; (2) the sensitivity towards Mn²⁺, methione, and other sulfur-containing amino acids and (3) their immunologic properties. CSAD II has been purified to about 2,500-fold by a combination of column chromatographies and polyacrylamide gel electrophoresis (PAGE). The purity of the enzyme preparation was established as judged from the following observations: (1) a single protein band was observed under various electrophoretic conditions, e.g., 5–20% nondenaturing PAGE, 7% nondenaturing PAGE and 10% SDS-PAGE and (2) in nondenaturing PAGE, the protein band comigrated with CSAD activity. CSAD II has a molecular weight of 90 kDa and is a homodimer consisting of two 43 ± 2 kDa subunits. CSAD appears to require Mn²⁺ for its maximum activity. Other divalent cations fail to replace Mn²⁺ in activation of CSAD activity. However, the precise role of Mn²⁺ in the action of CSAD remains to be determined.     

Characterization of phospholipase A2 activation by plasmin in cultured bovine endothelial cells

Treatment of cultured bovine carotid artery endothelial cells with 0.1 µM human plasmin has been reported to induce a receptor-mediated short burst of arachidonate release, which is a pertussis toxin-sensitive and extracellular calcium-dependent reaction. Plasmin-induced calcium influx in cells was significantly inhibited by pretreatment with pertussis toxin, indicating that the former was coupled with a pertussis toxin-sensitive guanosine 5-triphosphate (GTP)-binding protein. Plasmin significantly induced the formation of lysophosphatidylcholine but not lysophosphatidylethanolamine. A cellular phospholipase A₂ with an arachidonyl specificity at the sn-2 position of phosphatidylcholine, which required submicromolar calcium, was identified as a cytosolic phospholipase A₂ by immunoblot analysis. By a cell-free enzyme activity assay and immunoblot analysis, plasmin was found to induce a translocation of the cytosolic phospholipase A₂ from the cytosol to the membrane. Taken together, the results suggest that plasmin bound to its putative receptor and activated a GTP-binding protein coupled to calcium influx channel, followed by translocation and activation of cytosolic phospholipase A₂ in endothelial cells.     

The allometry of algal respiration

For 30 years, study after study has shown that respiration ratesincrease as {small tilde}0.75 of body size for organisms rangingfrom protozoans to mammals. However, a number of studies suggestedthat the respiration-size relationship for algae may be a rareexception to this general rule. Algal respiration may be almostproportional to cell size, such that the slope of the respiration-sizerelationship is closer to unity. The present study examinedthe effect of cell size and taxon on phytoplankton respiration,using data collected from the literature. To this end, we collecteda data set of 178 observations of algal respiration and cellsize representing six divisions-chlorophytes. chrysophytes,cyanophytes, euglenophytes, pyrrophytes and rhodophytes. Therelationship between respiration (R, in p1 O₂ cell^–1 h^–1)and cell carbon content (C, in pg C cell^–1) is describedas R = 0.030C^{0 93} and the exponent is significantly >3/4.When we expressed cell size in terms of volume, the exponentdecreased to 0.88 but this is still significantly >3/4. Amongthe six divisions studied, chlorophytes, euglenophytes and rhodophytesseemed to differ significantly in their respiration-size relationshipfrom other taxa. However, euglenophytes and rhodophytes havesuch small size ranges that no meaningful relationships canbe developed for those groups alone. The chlorophyte respiration-sizerelationship has obvious patterns in its residuals which mayindicate that significant sources of error were not controlledin these heterogeneous data. Thus, for the present, the generalmodel seems most appropriate for the prediction of respirationrates of phytoplankton.     

Regulated underexpression of the FliM protein of Escherichia coli and evidence for a location in the flagellar motor distinct from the MotA/MotB torque generators.

The FliM protein of Escherichia coli is essential for the assembly and function of flagella. Here, we report the effects of controlled low-level expression of FliM in a fliM null strain. Disruption of the fliM gene abolishes flagellation. Underexpression of FliM causes cells to produce comparatively few flagella, and most flagella built are defective, producing subnormal average torque and fluctuating rapidly in speed. The results imply that in a normal flagellar motor, multiple molecules of FliM are present and can function independently to some degree. The speed fluctuations indicate that stable operation requires most, possibly all, of the normal complement of FliM. Thus, the FliM subunits are not as fully independent as the motility proteins MotA and MotB characterized in earlier work, suggesting that FliM occupies a location in the motor distinct from the MotA/MotB torque generators. Several mutations in fliM previously reported to cause flagellar paralysis in Salmonella typhimurium (H. Sockett, S. Yamaguchi, M. Kihara, V.M. Irikura, and R. M. Macnab, J. Bacteriol. 174:793-806, 1992) were made and characterized in E. coli. These mutations did not cause flagellar paralysis in E. coli; their phenotypes were more complex and suggest that FliM is not directly involved in torque generation.     

Post-exercise metabolic rate in Atlantic cod and its dependence upon the method of exhaustion

This study tests whether or not post-exercise oxygen consumption rates ( M o₂) in fish are dependent upon how exhaustion is induced. A group of eight Atlantic cod ( Gadus morhua ) were each exercised using (1) a critical swimming speed ( U _crit) protocol, (2) an exercise protocol designed to measure anaerobic capacity of fish ( U _burst), and (3) a protocol in which the fish were chased to exhaustion manually. M o₂ was measured for a 2-h period following exhaustion induced by all three exercise regimes ( U _crit, U _burst and chase). Post-exercise M o₂ following exhaustion from the U _burst and chase protocols were significantly higher than post-exercise M o₂ following the U _crit protocol. Each fish during the U _crit protocol exhibited maximal M o₂ during exercise rather than during recovery, yet 75% of the fish during U _brust recovery and 100% during chase recovery exhibited M o₂ higher than that measured during U _crit exercise. These results, as well as the large interindividual variations in M o₂ among the eight fish, show that post-exhaustion M o₂ is specific to the exercise regime employed, thus, investigators must exercise caution when combining results from different exercise protocols and/or individuals.     

Modulation of insulin action by vanadate: evidence of a role for phosphotyrosine phosphatase activity to alter cellular signaling

A number of vanadium compounds (vanadate, vanadyl sulfate, metavanadate) have insulin-mimicking actions bothin vitro andin vivo. They have multiple biological effects in cultured cells and interact directly with various enzymes. The inhibitory action on phosphoprotein tyrosine phosphatases (PTPs) and enhancement of cellular tyrosine phosphorylation appear to be the most relevant to explain the ability to mimic insulin. We demonstrated that in rat adipocytes both acute insulin effects, e.g. stimulation of IGF-II and transferrin binding and a chronic effect, insulin receptor downregulation, were stimulated by vanadate. Vanadate also enhanced insulin binding, particularly at very low insulin concentrations, associated with increased receptor affinity. This resulted in increased adipocyte insulin sensitivity. Finally vanadate augmented the extent of activation of the insulin receptor kinase by submaximal insulin concentrations. This was associated with a prolongation of the insulin biological response, lipogenesis, after removal of hormone.In conclusion: in rat adipocytes vanadate promotes insulin action by three mechanisms, 1) a direct insulin-mimetic action, 2) an enhancement of insulin sensitivity and 3) a prolongation of insulin biological response. These data suggest that PTP inhibitors have potential as useful therapeutic agents in insulin-resistant and relatively insulin-deficient forms of diabetes mellitus.     

Bacteriophage T12 of Streptococcus pyogenes integrates into the gene encoding a serine tRNA

The region of temperate bacteriophage T12 responsible for integration into the chromosome of Streptococcus pyogenes has been identified. The integrase gene ( int ) and the phage attachment site ( attP ) are found immediately upstream of the gene for speA , the latter of which is known to be responsible for the production of erythrogenic toxin A (also known as pyrogenic exotoxin A). The integrase gene has a coding capacity for a protein of 41 457 Da, and the C-terminus of the deduced protein is similar to other conserved C-terminal regions typical of phage integrases. Upstream of int is a second open reading frame, which is capable of encoding an acidic protein of 72 amino acids (8744 Da); the position of this region in relation to int suggests it to be the phage excisionase gene ( xis ). The arms flanking the integrated prophage ( attL and attR ) were identified, allowing determination of the sequences of the phage ( attP ) and bacterial ( attB ) attachment sites. A fragment containing the integrase gene and attP was cloned into a streptococcal suicide vector; when introduced into S. pyogenes by electrotransformation, this plasmid stably integrated into the bacterial chromosome at attB . The insertion site for the phage into the S. pyogenes chromosome was found to be in the anticodon loop of a putative type II gene for a serine tRNA. attP and attB share a region of identity that is 96 bp in length; this region of identity corresponds to the 3' end of the tRNA gene such that the coding sequence remains intact after integration of the prophage. The symmetry of the core region of att may set this region apart from previously described phage attachment sites (Campbell, 1992), and may play a role in the biology of this medically important bacteriophage.