培养大鼠搏动心肌细胞在缺氧和复氧时的电生理观察

应用细胞内微电极技术记录到37个培养大鼠搏动心肌细胞充氮前后和复氧后的电活动参数。结果提示:充氮10min后,最大舒张电位(MDP),最大除极速度(V_(max)),动作电位振幅(APA)和动作电位时程(APD)等参数明显降低;自发节律增快,并出现多种形式的节律失常。83.8％细胞在充氮后30min内停搏,16.2％在50min左右停搏。复氧后,86.5％细胞在5min内复跳,13.5％未能复跳;12.5％复跳细胞在复跳10min内再次停搏。复跳细胞的各项电活动参数在30min内未能恢复到充氮前水平(p<0.05),且呈现不同程度的各类异常电活动。本结果对进一步研究心肌细胞缺氧和复氧损伤有一定意义。     

皮层第二体感区下行活动对尾核神经元伤害性反应的影响

本实验用清醒麻痹的家猫观察到,皮层第二体感区(SⅡ区)的下行活动对尾核痛相关单位的自发放电和伤害性反应均有影响。在35个痛相关单位中,刺激SⅡ区后,62.7％的自发放电受到影响(p<0.05)。在72个痛相关单位中,刺激SⅡ区后,77.8％的伤害性反应受到抑制(p<0.01),其中抑制痛兴奋单位的伤害性反应最为明显。而用利多卡因局部阻滞SⅡ区后,33个痛相关单位中,51.5％的伤害性反应增强(p<0.01)。本结果提示:(1)SⅡ区的下行活动不仅对尾核痛相关单位的自发活动有影响,而且对痛相关单位的伤害性反应有以抑制为主的下行性调制作用。(2)在正常机能状态下,SⅡ区对伤害性信息向尾核的传递有一定的紧张性的下行性抑制作用。     

Ⅱ型限制性核酸内切酶NcrⅠ的研究

 在肉色诺卡氏菌C-212株Nocardia carnea C-212中筛选到一种Ⅱ型限制性核酸内切酶NcrⅠ,经与BglⅡ的λDNA降解物的酶谱比较,以及酶识别特异性和切割位点的检测,证明了NcrⅠ是已知的限制酶BglⅡ的同切限制酶,而且其切割位点也与BglⅡ相同,其为:     

Quantitative analysis of the combined effects of temperature, evaporative demand and light on leaf elongation rate in well-watered field and laboratory-grown maize plants

The respective effects of meristem temperature, vapour pressuredeficit (VPD) and photosynthetic photon flux density (PPFD)on leaf elongation rate (LER) of maize, in the absence of waterdeficit in the soil have been quantified. This analysis wascarried out in a series of field experiments in northern andsouthern France over several seasons and years, and in growthchamber experiments. LER was measured with 10 min steps, togetherwith meristem temperature, VPD and PPFD at leaf level in threetypes of experiments: in growth chamber experiments with stepsin PPFD or VPD at constant meristem temperature, in growth chamberexperiments with several combinations of constant, but contrasting,PPFDs, VPDs and meristem temperatures, and in the field withfluctuating conditions, (i) When evaporative demand was low(night or day with low air VPD), LER was only linked to meristemtemperature, regardless of other climatic conditions, (ii) Lighthad no effect per se on LER in the range from 0 to 1500 molm^–2 s^–1 for time-scales longer than 2 h, providedthat its indirect effects on meristem temperature and on evaporativedemand were corrected (in the growth chamber) or taken intoaccount (in the field), and provided that cumulated PPFD overa weekly time-scale was compatible with field conditions, (iii)Evaporative demand sensed by growing leaves, as estimated bymeristem-to-air vapour pressure difference, markedly affectedLER in the range from 1–4 kPa, at all time-scales understudy, with a unique relationship in the growth chamber (constantconditions) and in the field (fluctuating conditions). Thiseffect was only observed when PPFD was high enough for stomatato open. The negative effect of evaporative demand on LER wasprobably not due to long distance root-to-shoot signalling,since soil was wet, calculated root water potential remainedclose to 0 MPa and concentration of ABA in the xylem sap wasvery low. Therefore, it is proposed to model maize LER witha two-step process, involving the calculation of the maximumLER at a given meristem temperature and then the calculationof the reduction in LER due to evaporative demand. Joint analysisof the whole set of data by using the two equations yieldeda r² of 0.75. This two-step process would be more accurate thanthe provision of LER from temperature only in cases where airVPD frequently exceeds 2 kPa. Key words: Leaf growth, light, evaporative demand, temperature, thermal time, water deficit, ABA, Zea mays L.     

Stationary phase-associated protein expression in Mycobacterium tuberculosis: function of the mycobacterial alpha-crystallin homolog.

The majority of active tuberculosis cases arise as a result of reactivation of latent organisms which are quiescent within the host. The ability of mycobacteria to survive extended periods without active replication is a complex process whose details await elucidation. We used two-dimensional gel electrophoresis to examine both steady-state protein composition and time-dependent protein synthetic profiles in aging cultures of virulent Mycobacterium tuberculosis. At least seven proteins were maximally synthesized 1 to 2 weeks following the end of log-phase growth. One of these proteins accumulated to become a predominant stationary-phase protein. N-terminal amino acid sequencing and immunoreactivity identified this protein as the 16-kDa alpha-crystallin-like small heat shock protein. The gene for this protein was shown to be limited to the slowly growing M. tuberculosis complex of organisms as assessed by Southern blotting. Overexpression of this protein in wild-type M. tuberculosis resulted in a slower decline in viability following the end of log-phase growth. Accumulation of this protein was observed in log-phase cultures following a shift to oxygen-limiting conditions but not by other external stimuli. The protein was purified to homogeneity from overexpressing M. smegmatis in two steps and shown to have a significant ability to suppress the thermal denaturation of alcohol dehydrogenase. Collectively, these results suggest that the mycobacterial alpha-crystallin protein may play a role in enhancing long-term protein stability and therefore long-term survival of M. tuberculosis.     

Distribution and breeding biology of offshore cichlids in Lake Malawi/ Niassa

Synopsis Lake Malawi/Niassa is the second largest rift valley lake in Africa, with an area of 28 800 km², and an average and maximum depth of 292 m and>700 m, respectively. The lake is well known for the great diversity of fish occurring in the inshore zone. However, the offshore fish community is poorly documented. To rectify this, regular sampling was undertaken over two years, using trawl and gillnets at six offshore locations. This paper reports on the species composition, spatial distribution and breeding biology of the dominant cichlids species from the offshore pelagic zone. Cichlids formed approximately 88% of the offshore fish biomass. Most abundant were two species of zooplanktivores in the genus Diplotaxodon that made up 71% of the offshore fish biomass. An undescribed species, given the cheironym D. bigeye, was mainly found at a depth of 220 m during the day, but moved into near surface waters at night when the moon was full. This species was absent from the shallow regions of the lake. The most abundant offshore species was D. limnothrissa, which was distributed evenly throughout the lake to depths of 220 m. A less common offshore zooplanktivore was Copadichromis quadrimaculatus that formed 5% of the biomass and was confined to the upper 100 m of the water column. The main piscivores were in the genus Rhamphochromis and formed approximately 10% of the offshore fish biomass. The two dominant taxa were R. longiceps and the large Rhamphochromis group, and both were more common in the southern half of the lake. The former occurred mainly in the upper 100 m of the water column and the latter mainly at depths of 100–150 m. The length at maturity and fecundity for the dominant offshore species were estimated and seasonal breeding cycles determined from gonad activity and gonado-somatic indices.     

肝再生的调控及原癌基因表达

肝脏再生过程中受到多种体液因素的调控。肝细胞生长因子（ＨＧＦ）、肝刺激因子（ＨＳＳ）等对肝细胞有促分裂作用;转化生长因子β１（ＴＧＦβ１）等则具有抑制作用。此外,肝再生还需要去甲肾上腺素（ＮＥ）、胰岛素等辅助分裂原的存在,共同调节肝再生。肝细胞分裂增殖与原癌基因表达密切相关。肝细胞从静息期进入细胞周期,以及在整个细胞周期中,某些原癌基因有特征性的表达。     

以异源四倍体体细胞杂种为父本杂交培育三倍体柑桔植株的研究

以 3个柑桔原生质体融合而来的四倍体体细胞杂种为父本 ,与二倍体单胚性种柚子 (Citrusgrandis)以及单多胚混合型品种“华农本地早”桔 (C.reticulata)有性杂交 ,授粉后 90 d,发现种子干瘪 ,大部分种子的胚败育。将干瘪种子在 MT附加 1mg/L GA3 或 50 0 mg/L麦芽浸出物的培养基中 ,经培养抢救 ,有 2 5.6%的种子萌发成苗或继续进行胚的生长 ,后者进一步诱导能形成丛芽 ,经试管嫁接或诱导生根形成完整植株。共获得 6个组合 73棵完整植株 ,染色体数检查表明 ,2 0株为三倍体 (2 n=3x=2 7) ,32株为二倍体 (2 n=2 x=18) ,8株为非整倍体 ,其它 13株还有待于进一步检查。     

In vitro plantlet regeneration from cotyledon, hypocotyl and root explants of hybrid seed geranium

In vitro plantlet regeneration systems for the seed geranium (Pelargonium x hortorum Bailey) using cotyledon, hypocotyl and root explants were optimized by studying the influence of seedling age, growth regulators and excision orientation on organogenesis. Indole-3-acetic acid combined with zeatin yielded the highest rate of shoot production on cotyledon explants (0.2–2 shoots per explant). More shoots were produced on explants cut from the most basal region of cotyledons from 2 to 4-day-old seedlings than from older seedlings or more distal cut sites. Hypocotyl explants produced the highest number of shoots, up to 40 shoots per explant, on indole-3-acetic acid (2.8–5.6 mM) + zeatin (4.6 mM) or thidiazuron (4.5 mM). Maximum shoot formation (0.3–1.4 shoots per explant) on root explants occurred when they were cultured on medium containing zeatin. Regenerated shoots rooted best on a basal medium containing no growth regulators. There were substantial differences among cultivars in shoot formation from each of the explant systems.Abbreviations BA 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - NAA naphthaleneacetic acid - TDZ thidiazuron     

Acquisition and assimilation of gaseous ammonia as revealed by intracellular pH changes in leaves of higher plants

Atmospheric ammonia (NH₃) from various anthropogenic sources has become a serious problem for natural vegetation. Ammonia not only causes changes in plant nitrogen metabolism, but also affects the acid-base balance of plants. Using the pH-sensitive fluorescent dyes pyranine and esculin, cytosolic and vacuolar pH changes were measured in leaves of C₃ and C₄ plants exposed for brief periods to concentrations of NH₃ in air ranging from 1.33 to 8.29 mol NH₃ · mol^-1 gas (0.94–5.86 mg · m^-3). After a lag phase, uptake of NH₃ from air at a rate of 200 nmol NH₃ · m ^{- 2} leaf area · s^{- 1} into leaves of Zea mays L. increased pyranine fluorescence indicating cytosolic alkalinisation. The increase was much larger in the dark than in the light. In illuminated leaves of the C₃ plant Pelargonium zonale L. and the C₄ plants Z. mays and Amaranthus caudatus L., NH₃-dependent cytosolic alkalinisation was particularly pronounced when CO₂ was supplied at very low levels (16 or 20 mol CO₂ · mol^{- 1} gas, containing 210 mmol O₂ · mol^{- 1} gas). An increase in esculin fluorescence, which was smaller than that of pyranine, was indicative of trapping of some of the NH₃ in the vacuoles of leaves of Spinacia oleracea L. and Z. mays. Photosynthesis and transpiration remained unchanged during exposure of illuminated leaves to NH₃, yielding an influx of 200 nmol NH₃ · m^-2 leaf area · s^-1 for up to 30 min, the longest exposure time used. Both CO₂ and O₂ influenced the extent of cytosolic alkalinisation. At 500 mol CO₂ · mol^-1 gas the cytosolic alkalinisation was suppressed more than at 16 or 20 mol CO₂ · mol^-1 gas. The suppressing effect of CO₂ on the NH₃induced alkalinisation was larger in illuminated leaves of the C₄ plants Z. mays and A. caudatus than in leaves of the C₃ plant P. zonale. A reduction of the O₂ concentration from 210 to 10 mmol O₂ · mol ^-1 gas, which inhibits photorespiration, increased the NH₃induced cytosolic alkalinisation in C₃ plants. Suppression by CO₂ or O₂ of the alkaline pH shift caused by the dissolution and protonation of NH₃ in queous leaf compartments, and possibly by the production of organic compounds synthesised from atmospheric NH₃, indicates that NH₃ which enters leaves is rapidly assimilated if photosynthesis or photorespiration provide nitrogen acceptor molecules.This work was supported by the Biotechnology and Biological Sciences Research Council and the Deutsche Forschungsgemein-schaft within the framework of the research of Sonderforschun-gsbreich 251 of the University of Würzburg. We are grateful to Dr. B. Wollenweber (The Royal Veterinary and Agricultural University, Denmark) for discussions.