K+ channels of stomatal guard cells: Abscisic-acid-evoked control of the outward rectifier mediated by cytoplasmic pH

The activation by abscisic acid (ABA) of current through outward-rectifying K⁺ channels and its dependence on cytoplasmic pH (pH_i) was examined in stomatal guard cells of Vicia faba L. Intact guard cells were impaled with multibarrelled and H⁺-selective microelectrodes to record membrane potentials and pH_i during exposures to ABA and the weak acid butyrate. Potassium channel currents were monitored under voltage clamp and, in some experiments, guard cells were loaded with pH buffers by iontophoresis to suppress changes in pH_i. Following impalements, stable pH_i values ranged between 7.53 and 7.81 (7.67±0.04, n = 17). On adding 20 M ABA, pH_i rose over periods of 5–8 min to values 0.27±0.03 pH units above the pH_i before ABA addition, and declined slowly thereafter. Concurrent voltage-clamp measurements showed a parallel rise in the outward-rectifying K⁺ channel current (I_{K, out}) and, once evoked, both pH_i and I_{K, out} responses were unaffected by ABA washout. Acid loads, imposed with external butyrate, abolished the ABA-evoked rise in I_{K, out}. Butyrate concentrations of 10 and 30 mM (pH₀ 6.1) caused pH_i to fall to values near 7.0 and below, both before and after adding ABA, consistent with a cytoplasmic buffer capacity of 128±12 mM per pH unit (n = 10) near neutrality. Butyrate washout was characterised by an appreciable alkaline overshoot in pH_i and concomitant swell in the steady-state conductance of I_{K, out}. The rise in pH_i and i_{K, out} in ABA were also virtually eliminated when guard cells were first loaded with pH buffers to raise the cytoplasmic buffer capacity four- to sixfold; however, buffer loading was without appreciable effect on the ABA-evoked inactivation of a second, inward-rectifying class of K⁺ channels (I_{K, in}). The pH_i dependence of I_{K, out} was consistent with a cooperative binding of at least 2H⁺ (apparent pK_a = 8.3) to achieve a voltage-independent block of the channel. These results establish a causal link previously implicated between cytoplasmic alkalinisation and the activation of I_{K, out} in ABA and, thus, affirm a role for H⁺ in signalling and transport control in plants distinct from its function as a substrate in H⁺-coupled transport. Additional evidence implicates a coordinate control of I_{K, in} by cytoplasmic-free [Ca²⁺] and pH_i.Abbreviations ABA abscisic acid - [Ca²⁺]_i cytoplasmic free [Ca²⁺]_i - E_K K⁺ equilibrium potential - I_{K, out}, I_{K, in} outward-, inward-rectifying K⁺ channel (current) - I-V current-voltage (relation) - Mes 2-(N-morpholino)ethanesulfonic acid - pH_i cytoplasmic pH - Tes 2-{[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]-amino}ethanesulfonic acid - V_m membrane potential We are grateful to G. Thiel (Pflanzenphysiologisches Institut, Universität Göttingen, Germany) for helpful discussions. This work was possible with equipment grants-in-aid from the Gatsby Charitable Foundation, the Royal Society and the University of London Central Research Fund. F.A. holds a Sainsbury Studentship.     

超声引导下神经阻滞麻醉联合全身麻醉对股骨头置换老年患者麻醉效果及术后认知功能的影响

摘要 目的：研究超声引导神经阻滞麻醉联合全身麻醉对股骨头置换老年患者的麻醉效果及对术后认知功能的影响。方法：选取2019年1月至2021年12月期间我院收治的150例拟行股骨头置换术的老年患者,随机分为对照组和观察组,各75例。对照组患者行常规全身麻醉,观察组在对照组的基础上行超声引导下神经阻滞麻醉,比较两组患者的血流动力学指标,拔管时间、苏醒时间和复苏室停留时间,苏醒后疼痛,术后认知功能及不良反应的发生率。结果：观察组患者各时间点的心率（HR）和平均动脉压（MAP）均较对照组低（P＜0.05）。观察组拔管时间、苏醒时间和复苏室停留时间均较对照组短（P＜0.05）。观察组患者术后视觉模拟评分（VAS）均低于同一时间点的对照组（P＜0.05）。与对照组相比,观察组患者术后1 h、12 h和24 h的简易智力状态检查（MMSE）评分均较高（P＜0.05）,观察组术后1 h、12 h和24 h术后认知功能障碍（POCD）发生率较对照组低（P＜0.05）。两组患者不良反应发生率无差异（P＞0.05）。结论：超声引导神经阻滞麻醉可稳定血流动力学,缩短拔管、苏醒及复苏室停留时间,减轻术后疼痛,改善术后认知功能,减少POCD的发生,值得临床推广。     

不同容量胸段经椎间孔硬膜外注射阻滞范围的CT影像研究及诊断性阻滞的可行性分析

摘要 目的：探讨不同容量对胸段经椎间孔硬膜外注射（TFEI）药液扩散范围和镇痛效果的影响以及胸段TFEI用于诊断性阻滞的可行性。方法：选择2021年1月至2022年12月南京大学医学院附属鼓楼医院收治的胸段带状疱疹相关疼痛患者140例,随机分为4组,实施单次TFEI并分别注入不同容量含造影剂局麻药（A组：0.2 mL;B组：0.5 mL;C组：1.0 mL;D组：2 mL）,CT扫描并观察造影剂在硬膜外向头侧、尾侧及总扩散节段,造影剂在椎间孔、同侧椎旁间隙、同侧及对侧硬膜外间隙扩散情况,判断是否为选择性神经根阻滞。评估注射前、注射后30分钟及24小时视觉模拟评分（VAS）。结果：头侧和尾侧扩散节段以及总扩散节段数,D组最多,A组最少（P<0.05）;C组、D组造影剂扩散≥3个节段发生率明显高于B组（P<0.05）;C组、D组病例造影剂扩散至同侧椎旁间隙和对侧硬膜外间隙的发生率明显高于A组、B组（P<0.05）,仅A组37.1%的病例实现选择性神经根阻滞,其余各组均无选择性阻滞病例。注射后30分钟,C、D组VAS评分显著低于A、B组（P<0.05）;注射后24小时,D组VAS评分显著低于A、B组（P<0.05）。结论：胸段带状疱疹相关疼痛患者TFEI药液扩散范围随注射容量的增加而扩大,且在硬膜外倾向于头侧扩散,2 mL容量单次TFEI可阻滞3个以上的神经节段,获得良好的镇痛效果。胸段TFEI行诊断性阻滞的可行性较差。     

加巴喷丁联合射频脉冲、神经阻滞治疗带状疱疹后神经痛的效果

摘要 目的：探讨加巴喷丁联合脉冲射频、神经阻滞治疗带状疱疹后遗神经痛的效果。方法：本研究选取106例确诊为带状疱疹后遗神经痛的患者,采用随机数表法将其分为对照组A（36例）、试验组B（35例）和试验组C（35例）。A组患者采用口服加巴喷丁进行治疗,B组患者采用加巴喷丁联合神经阻滞治疗,C组患者采用加巴喷丁联合脉冲射频治疗,观察比较三组患者的治疗效果,分别对视觉模拟评分（VAS）、夜间睡眠评分（SRSS）、疼痛程度（NRS）、不良反应及综合疗效进行统计评估。结果：治疗3天、7天、14天和1月后,与对照组A比较,B、C组患者的VAS、SRSS和NRS评分较治疗前均有显著降低（P<0.05）;试验组B、C不良反应发生率分别为8.57%（3/35）和5.71%（2/35）,对照组A不良反应发生率为22.22%（8/36）,试验组不良反应发生率显著低于对照组,差异具有统计学意义（P<0.05）。结论：相较于使用加巴喷丁和加巴喷丁联合神经阻滞这两种治疗方案,加巴喷丁联合射频脉冲及神经阻滞治疗带状疱疹后遗神经痛能够在短期内有效缓解患者疼痛并改善睡眠状况。     

不同剂量右美托咪定联合超声引导下多点髂筋膜间隙阻滞在老年全髋关节置换术中的应用效果分析

摘要 目的：探讨超声引导下多点髂筋膜间隙阻滞联合不同剂量右美托咪定在老年全髋关节置换术（THA）中的应用效果。方法：选取2019年11月-2022年12月北京积水潭医院贵州医院择期行THA的老年患者120例,按照随机数字表法分为A组（n=40）、B组（n=40）、C组（n=40）,三组均接受超声引导下多点髂筋膜间隙阻滞治疗,在此基础上,A组、B组、C组分别给予0.5 μg/kg、1.0 μg/kg、1.5 μg/kg右美托咪定注射液。对比三组临床指标、血流动力学指标、应激反应指标、脑神经损伤情况,观察三组围术期间不良反应发生情况。结果：B组、C组的阻滞消退时间、拔管时间长于A组,首次下床时间短于A组（P<0.05）。B组、C组的阻滞消退时间、拔管时间、首次下床时间对比无差异（P>0.05）。三组麻醉诱导后即刻（T1）~手术结束时（T3）时间点心率（HR）、平均动脉压（MAP）升高后降低（P<0.05）。B组、C组的T1~T3时间点HR、MAP均低于A组,且B组低于C组（P<0.05）。三组T3时间点皮质醇（Cor）、白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）升高（P<0.05）。B组、C组的T3时间点Cor、IL-6、TNF-α均低于A组,且B组低于C组（P<0.05）。B组、C组的T3时间点中枢神经特异性蛋白（S100β）均低于A组,且B组低于C组（P<0.05）。B组、C组的T3时间点脑源性神经生长因子（BDNF）均高于A组,且B组高于C组（P<0.05）。三组不良反应发生率组间对比无差异（P>0.05）。结论：超声引导下多点髂筋膜间隙阻滞联合1.0 μg/kg右美托咪定在THA中具有更好的麻醉效果,可缩短首次下床时间,减轻机体应激反应、血流动力学波动以及脑部神经损伤。     

A Wuchiapingian (Late Permian) brachiopod fauna from an exotic block in the Indus–Tsangpo suture zone, southern Tibet, and its palaeobiogeographical and tectonic implications

A brachiopod fauna including 19 species of 17 genera from an exotic block in the Indus–Tsangpo suture zone in southern Tibet is described and illustrated. The brachiopod fauna is dominated by Martinia elegans and two new taxa: Jinomarginifera lhazeensis gen. et sp. nov. and Zhejiangospirifer giganteus sp. nov. The fauna is closely comparable with those from the middle and upper parts of the Wargal Formation and the Chhidru Formation in the Salt Range of Pakistan, the Chitichun Limestone in southern Tibet, and the Basleo area of West Timor, and these correlations suggest a Wuchiapingian age. The fauna exhibits substantial links with both peri–Gondwanan and Cathaysian faunas, which may imply that it is a seamount biota originally located in the southern margin of the Neotethys during the Late Permian, and was later (in the early Cenozoic) displaced and became sandwiched into younger marine deposits in the collision process between India and Eurasia.     

Inward rectifier potassium channels in plants differ from their animal counterparts in response to voltage and channel modulators

We have investigated the electrophysiological basis of potassium inward rectification of the KAT1 gene product from Arabidopsis thaliana expressed in Xenopus oocytes and of functionally related K⁺ channels in the plasma membrane of guard and root cells from Vicia faba and Zea mays. The whole-cell currents passed by these channels activate, following steps to membrane potentials more negative than –100 mV, with half activation times of tens of milliseconds. This voltage dependence was unaffected by the removal of cytoplasmic magnesium. Consequently, unlike inward rectifier channels of animals, inward rectification of plant potassium channels is an intrinsic property of the channel protein itself. We also found that the activation kinetics of KAT1 were modulated by external pH. Decreasing the pH in the range 8.5 to 4.5 hastened activation and shifted the steady state activation curve by 19 mV per pH unit. This indicates that the activity of these K⁺ channels and the activity of the plasma membrane H⁺-ATPase may not only be coordinated by membrane potential but also by pH. The instantaneous current-voltage relationship, on the other hand, did not depend on pH, indicating that H⁺ do not block the channel. In addition to sensitivity towards protons, the channels showed a high affinity voltage dependent block in the presence of cesium, but were less sensitive to barium. Recordings from membrane patches of KAT1 injected oocytes in symmetric, Mg²⁺-free, 100 mM-K⁺, solutions allowed measurements of the current-voltage relation of single open KAT1 channels with a unitary conductance of 5 pS. We conclude that the inward rectification of the currents mediated by the KAT1 gene product, or the related endogenous channels of plant cells, results from voltage-modulated structural changes within the channel proteins. The voltage-sensing or the gating-structures appear to interact with a titratable acidic residue exposed to the extracellular medium. Correspondence to: R. Hedrich     

Genetic control of chalcone isomerase activity in anthers of Petunia hybrida

The gene Po in pollen of Petunia hybrida Vilm. controls a discrete step in flavonoid biosynthesis. In recessive genotypes, naringenin-chalcone (4, 2,4,6-tetrahydroxychalcone) is accumulated, whereas, under the influence of the wild-type allele flavonols and anthocyanins are formed. Enzymic investigations on anthers of four genetically defined lines with different pollen colouration revealed a clear correlation between accumulation of naringenin-chalcone and deficiency of chalcone isomerase (EC 5.5.1.6). The results allow the conclusion that chalcone is the first product of the flavanone synthase reaction in anthers of Petunia hybrida and that chalcone isomerase is essential for the formation of flavonols and anthocyanins. These results were similar to those previously obtained with Callistephus chinensis (L.) Nees.Abbreviations EGME ethylen glycol monomethyl ether - MeOH methanol - CI chalcone isomerase - HOAc acetic acid - TLC thinlayer chromatography     

Differential response of DPI-modified cardiac Na+ channels to antiarrhythmic drugs: No flicker blockade by lidocaine

Summary Elementary Na⁺ currents were recorded in cell attached patches from short-time cultured neonatal cardiocytes in order to test the hypothesis whether the open state of DPI-modified, noninactivating cardiac Na⁺ channels is basically sensitive to blocking drug molecules such as antiarrhythmics.Lidocaine (300 mol/liter) effectively reduced the open probability of cardiac Na⁺ channels and, at a stimulation rate of 1 Hz, depressed the reconstructed macroscopic peak I _Na to 40+ 3.5% of the predrug value. The same drug concentration failed to influence DPI-modified Na⁺ channels. Their open state proved almost insensitive to lidocaine. _open decreased only slightly to 85 ±2%. Still more importantly, the number of transitions between the conducting and a nonconducting configuration did not increase. At –40 mV, lidocaine may interfere with the open state with an association rate constant of 1.3×10⁵ mol^–1sec^–1 which is about two orders of magnitude smaller than the rate constant obtained with propafenone or prajmalium. Moreover, propafenone (10–20 mol/liter) or prajmalium (30 mol/liter) led to a tremendous increase in the number of transitions between the open and a nonconducting configuration. Lidocaine also failed to evoke a fast flicker blockade with reaction kinetics in the microsecond range.It is concluded that DPI-modified cardiac Na⁺ channels discriminate between lidocaine and other antiarrhythmic drugs. As a tentative explanation, this might be indicative for multiple binding sites for those drugs in cardiac Na⁺ channels.This work was supported by a grant from the Deutsche Forschungs-gemeinschagt (Ko 778/2–3), Bonn.