A double sequential immunofluorescence method demonstrating the co-localization of urotensins I and II in the caudal neurosecretory system of the teleost,Gillichthys mirabilis

Summary A double immunofluorescence method was devised to localize simultaneously urotensin-I (UI) and -II (UII) immunoreactivities in the caudal neurosecretory system of the goby, Gillichthys mirabilis. In a sequential fashion, sections of the posterior spinal cord and urophysis were treated with antiserum to corticotropin-releasing factor (CRF) that cross-reacts with UI, fluorescein-conjugated sheep anti-rabbit IgG, biotinylated anti-UII and rhodamine-conjugated avidin. UI and UII immunoreactivities appeared to coexist in some neurons and in most fibers and urophysial tissue; the remainder of the fibers and urophysis and the majority of neurons were immunoreactive for CRF/ UI only. No convincing evidence of immunoreactivity for UII only was found. A few nonreactive cells were seen, but these may not be neurosecretory neurons. The two immunoreactive cell types were not segregated topographically, and the intensity of perikaryal immunofluorescence for CRF/UI was variable. To explain these results a hypothesis that all caudal neurosecretory cells may synthesize both UI and UII and that immunoreactive differences may reflect different states of cellular activity, is suggested. This sequential double immunofluorescence method offers several advantages over other techniques and is especially useful for co-localization studies when primary antisera from different species are not available.     

Voltage-dependent depolarization of bacterial membranes and artificial lipid bilayers by the peptide antibiotic nisin

The peptide antibiotic nisin is shown to disrupt valinomycin-induced potassium diffusion potentials imposed on intact cells of Staphylococcus cohnii 22. Membrane depolarization occurred rapidly at high diffusion potentials while at low potentials nisin-induced depolarization was slower suggesting that nisin requires a membrane potential for activity. This assumption was proven in experiments with planar lipid bilayers (black lipid membranes). Macroscopic conductivity measurements indicated a voltage-dependent action of nisin. The potential must have a trans-negative orientation with respect to the addition of nisin (added to the cis-side) and a sufficient magnitude (ca. -100 mV). With intact cells the threshold potential was lower (-50 to -80 mV at pH 7.5 and below -50 mV at pH 5.5). Single channel recordings resolved transient multistate pores, strongly resembling those introduced by melittin into artificial bilayers. The pores had diameters in the range of 0.2–1 nm, and lifetimes of few to several hundred milliseconds. The results indicate that nisin has to be regarded as a membrane-depolarizing agent which acts in a voltage-dependent fashion.Abbreviations BLM Black lipid membranes - CCCP carbonyl cyanide m-chlorophenylhydrazone - DOPC dioleoyl phosphatidylcholine - PS phosphatidylserine - TPP⁺ tetraphenylphosphonium cation     

Cloning of a Vero toxin (VT2) gene from a VT2-converting phage isolated from Escherichia coli 0157: H7

Abstract A Vero toxin (VT2 or Shiga-like toxin II)-converting phage was isolated from Escherichia coli 0157: H7 strain J-2. Nontoxigenic E. coli C600 produced VT2 when lysogenized with the toxin-converting phage. Eco RI fragments of the phage DNA were ligated with Eco RI-digested pBR322 or pUC118 and were transformed into E. coli MC1061 or MV1184. Transformants exhibiting VT2 production commonly contained a 4.6 kb Eco RI fragment. It was found that a 2.3 kb Kpn I- Sph I fragment coded VT2 production and that this fragment hybridized weakly with the 2.1 kb fragment encoding VT1.     

Neurochemical evidence for two types of presynaptic alpha2-adrenoceptors

Neurochemical and pharmacological evidence has been obtained that noradrenergic varicosities (in mouse and rat vas deferens) and cholinergic varicosities (in the Auerbach's plexus) contain heterogenous alpha₂-adrenoceptors through which the release of [³H]noradrenaline and [³H]acetylcholine can be modulated. The quantitative data also support the hypothesis that different noradrenaline and xylazine sensitive alpha₂-adrenoceptors are present prejunctionally in the vas deferens and Auerbach's plexus preparations. Prazosin, although it has a presynaptic inhibitory effect on alpha₂-adrenoceptors of noradrenergic axon terminals, has no effect on cholinergic axon terminals. These data suggest that there are two different types of alpha₂-adrenoceptors at the presynaptic axon terminals.Special Issue Dedicated to Dr. Abel Lajtha     

利他林对戊巴比妥钠深麻醉家兔呼吸的兴奋效应及机制

在125只呼吸受戊巴比妥钠抑制并切断双侧颈迷走神经的家兔,分析了利他林(MP)呼吸效应的中枢部位及可能的递质。静脉注射 MP(2mg/kg)引起呼气时程显著缩短和呼吸频率显著增加。去除颈动脉体不影响 MP 兴奋呼吸的效应。将浸有2%MP 溶液的滤纸(2×2mm~2)贴敷在第四脑室底闩附近,或微量注射2%MP 2μl 到延髓 NTS 区,均可引起上述效应,但 NA区、NPBM 区和皮层体感运动区则无效。第四脑室或 NTS 区微量注射酚妥拉明均可阻断 MP兴奋呼吸的效应,但第四脑室注射心得安却不能阻断。结果提示,MP 可能作用于 NTS 区,在肾上腺素能α受体的参与下,受戊巴比妥钠抑制的家兔发生呼吸频率增快的效应。     

氟碳乳剂与右旋糖酐对犬缺血心肌侧支循环供氧的影响

本实验在14只麻醉开胸狗心脏上观察了氟碳乳剂与右旋糖酐稀释血液对心肌耗氧量与供应缺血心肌氧量关系的影响。以左室压力-时间指数(SPTI)作为心肌耗氧量的指标,根据冠脉有效侧支血流量(ECF)、PaO_2和 Hb 浓度计算供应缺血心肌的氧量。实验结果表明,低分子右旋糖酐稀释血液后,SPTI 暂时性轻度增加(稀释后30min 时较对照增加7.1±2.7%,P<0.05,稀释后60min 时增加2.8±1.2%,P>0.05),ECF 明显增多(稀释后30min 时较对照增加58.5±6.1%,P<0.01),缺血区边缘心肌氧供需关系未发生明显变化。氟碳乳剂稀释血液后,SPTI 的变化规律与右旋糖酐稀释后相同(稀释后30min 和60min 时分别较对照增加2.5±0.7%和1.9±0.8%)ECF 和 PaO_2升高(稀释后30min 时分别较对照增加53.9±6.7%和93±8.9%),供应缺血心肌的氧量显著增加,缺血区边缘心肌氧供需矛盾明显改善。     

柚花头香化学成分的研究(一)

本文首次报道柚花的香气成分。作者利用憎水性树脂XAD-4吸附柚鲜花的头香,并以毛细管气相色谱和毛细管气相色谱-质谱-计算机联用方法研究头香的化学组分,分离鉴定了17个已知化学成分。它们是芳樟醇、β-蒎烯、β-水芹烯、橙花叔醇等。     

Complete maps of IS1, IS2, IS3, IS4, IS5, IS30 and IS150 locations in Escherichia coli K12

Summary In this paper complete distribution maps are presented of the seven IS elements 1, 2, 3, 4, 5, 30 and 150. These maps were obtained during the construction of an almost complete restriction map of the Escherichia coli genome of K12 strain BHB2600. The positions of IS elements were correlated to this map. The distribution of integration sites of all IS types is nonrandom. Besides a large gap from 79 min to 96 min, there is a pronounced IS cluster at 6 min and another at 97 min, map locations that have low gene incidences on the classical map. One cluster coincides with a region of IS induced rearrangements. The IS distribution pattern was compared to patterns of strains W3110 and HB101.