外源氯胁迫对甘薯幼苗光合特性的影响

采用温室水培方法,以甘薯品种‘徐薯22’,‘苏薯11’和‘宁紫1号’为材料,外源氯(0、42.2、84.4、168.8、211mmol.L-1)胁迫处理7d、14d和21d,测定其生理生化及气体交换参数,探讨Cl-对甘薯幼苗生长以及光合特性的影响。结果表明:(1)‘徐薯22’叶面积与主蔓生长速率受外源氯胁迫而降低但仍维持在一定水平,而‘苏薯11’和‘宁紫1号’则在低浓度Cl-(≤84.4mmol.L-1)处理下高于对照。(2)3个甘薯品种叶片净光合速率(Pn)及光合色素含量在42.2mmol.L-1 Cl-处理下均高于对照;导致‘徐薯22’光合能力降低的主要因素为非气孔限制,而‘苏薯11’和‘宁紫1号’兼有气孔限制和非气孔限制因素,在短期或低浓度Cl-处理条件下以气孔限制因素为主。(3)处理21d时,甘薯幼苗根系氯离子含量低于14d且氯离子向地上部运输比率(SCl)上升;‘徐薯22’积累Cl-含量低于‘苏薯11’和‘宁紫1号’。研究发现:外源氯低浓度短期处理利于甘薯幼苗生长及光合作用的进行;在胁迫初期甘薯可有效抑制Cl-向地上部转运,但在处理第3周该抗逆性明显减弱;甘薯对氯胁迫的耐受性存在基因型差异,本实验中以‘徐薯22’耐性较强。     

睫状体色素上皮细胞容积激活性氯电流

为研究睫状体色素上皮 (pigmentedciliaryepithelial,PCE)细胞容积激活性Cl-电流的特性 ,用膜片箝全细胞记录技术记录了猪的低渗液诱发的容积激活性Cl-电流。此电流外向占优势 ,几乎没有时间依赖性失活 ,电流 电压曲线显示此电流反转电位 (- 6 3± 0 5mV)很接近氯离子平衡电位的计算值 (ECl=0mV)。电流的激活依赖于细胞内ATP ,细胞外ATP抑制外向电流和内向电流 ,但外向电流抑制率大于内向电流抑制率 (92 %比 74% ,P <0 0 1)。氯离子通道阻断剂tamoxifen抑制外向电流和内向电流 ,两个抑制率几乎相等 (85 %比 87% ,P >0 0 5 )。此电流特性与其他类型细胞的P糖蛋白相关电流很相似。结果提示PCE细胞容积激活性Cl-电流的形成可能与P糖蛋白有关     

长期施用含硫和含氯化肥对稻田杂草生长动态的影响

利用湖南祁阳红壤稻田长期定位试验,研究了在等量氮磷钾养分条件下,长期施用含Cl-、SO2-4和Cl-+SO2-4化肥水稻生育期间杂草种类和生物量的变化.结果表明:施肥34年后,施用含Cl-化肥处理水稻生育期间杂草的种类最多、总生物量(浮生杂草和湿生杂草的生物量之和)最大,早稻期间杂草平均总干物质量分别比含SO2-4和Cl-+SO2-4化肥处理增加了51.4%和17.6%,晚稻期间分别增加了144%和242%.含SO2-4和Cl-+SO2-4化肥处理稻田中浮生杂草生物量较大,而含Cl-化肥处理田间几乎没有浮生杂草生长.杂草总干物质量和湿生杂草干物质量均与土壤Cl-含量呈显著正相关(相关系数分别为0.764**和0.948**),与土壤SO2-4-S含量呈显著负相关(相关系数分别为0.849**和0.641*).土壤碱解氮和有效磷受土壤SO2-4-S、Cl-及pH的共同作用对杂草总干物质量产生影响.通过各种施肥措施维持土壤适宜pH及碱解氮、有效磷含量,提高土壤SO2-4-S含量、降低Cl-含量,能有效抑制南方红壤稻田中湿生杂草的生长,降低杂草总生物量.     

NaCl胁迫对坪山柚、福橘实生苗矿质营养吸收特性的影响

通过沙培和水培实验,采用原子吸收分光光度计法和离子吸收动力学方法,对NaCl胁迫下坪山柚(Citrus grandis Osbeck 'Pingshanyou')和福橘(C. reticulata Blanco 'Fuju')实生苗矿质营养吸收特性进行研究. 结果表明, 随NaCl浓度增加, 坪山柚和福橘幼苗地上部及根部Na 、 Cl-含量明显增加, K 、 Ca2 含量降低. 相同浓度NaCl胁迫下, 坪山柚地上部及根部Na 、 Cl-含量分别低于福橘, 而根部K 、 Ca2 含量及地上部K 含量、 K /Na 值、 Ca2 /Na 值均高于福橘.NaCl胁迫下,坪山柚、福橘幼苗Na 吸收量高于Cl-吸收量,Na 及 Cl-的吸收速率均随NaCl浓度的增大而增加.相同浓度NaCl胁迫下,坪山柚对Na 、Cl-的吸收速率低于福橘,这与坪山柚对Na 、Cl-的吸收具有较大的Km值及较小的Vmax值一致,表明坪山柚幼苗比福橘幼苗耐盐性强.     

外源氯对库拉索芦荟生长及品质的影响

通过4个水平的砂培试验研究了外源氯对库拉索芦荟(Aloe vera)生长和品质的影响。结果表明:(1)根系相对干质量的含水量、植株干质量、根冠比分别在施氯浓度≥25mmol·L-1、≥50mmol·L-1、≥100mmol·L-1时显著下降(P<0.05);(2)外源氯胁迫下,Cl-主要在芦荟根部积累,且外源氯浓度越高,根系Cl-含量越多而叶片的Cl-含量越少;(3)硝酸盐主要分布在根系和功能叶(5叶)上,施氯≤50mmol·L-1时叶片和根系的硝酸盐含量都显著低于对照;(4)随氯浓度水平的升高,蒽醌和可溶性蛋白含量均显著增加,游离氨基酸含量变化不大,芦荟甙含量在施氯≤50mmol·L时显著下降。     

阴离子对球茎茴香生长和精油含量的影响

分别用 6 0mmol·L-1浓度的HCO-3 、Cl-、SO2 -4复合盐处理定植后的球茎茴香 ,结果Cl-对生长的影响不明显 ,HCO-3 和SO2 -4处理的生长略差些 ,但植株中Na 、Ca2 、Mg2 含量均增加 ,全氮、全磷和K 的含量则都下降。精油含量以Cl-处理的为最高 ,SO2 -4的次之 ,HCO-3 的无变化。Cl-和SO2 -4的精油含量增加主要是烯的增加     

盐胁迫下水稻叶绿体中Na~ 、Cl~-积累导致叶片净光合速率下降

研究了 0～ 2 0 0mmol/L的NaCl胁迫下耐盐性不同的水稻品种Pokkali(耐盐 )和Peta(盐敏感 )根系、叶片和叶绿体中Na 、K 和Cl-含量的变化及其与叶片光合作用的关系。结果表明 :随着NaCl胁迫时间和浓度的增加 ,供试 2个品种在根、叶片和叶绿体中Na 、Cl-含量增加 ,K 含量下降。耐盐品种体内Na 、Cl-含量增加或K 含量减少的幅度小于盐敏感品种。在 2 0 0mmol/L的NaCl胁迫下盐敏感品种根、叶片和叶绿体中的Na /K 分别是耐盐品种的 2 0 8%、30 8%和 2 97%。与Na 相比 ,耐盐品种根系对K 的吸收和向叶片运输的选择性 (SK ,Na)较强。但在经过0、10 0和 2 0 0mmol/L的NaCl处理后 2个品种叶绿体中的Na /K 均高于叶片 (SK ,Na均小于 1)。盐胁迫下水稻叶绿体中Na 、Cl-含量和Na /K 与叶片净光合速率呈极显著负相关。     

盐胁迫下沙棘的渗透调节效应

分别用含有0、100、200和300mmol LNaCl的Hoagland培养液处理1年生沙棘(HippophaerhamnoidesL.)苗30d后,测定其鲜重,干重,含水量,可溶性糖、脯氨酸和无机离子(Na 、Cl-)的含量及叶片渗透势和渗透调节能力。结果表明:100mmol LNaCl处理的沙棘地上部和根的鲜重和干重最大,其含水量也最大;NaCl浓度超过100mmol L时,沙棘地上部分和根的鲜重和干重随盐浓度增加而逐步下降,其下降的趋势为地上部大于根部。随NaCl浓度不断升高,沙棘体内Na 和Cl-浓度随之升高,茎叶和根系中Cl-含量明显高于Na ,对Na 的相对吸收量多于Cl-。沙棘对盐胁迫有一定的适应能力,随NaCl浓度的升高,沙棘叶内脯氨酸含量升高,可溶性糖含量增加,渗透势降低,渗透调节能力增强。本结果可为盐碱地营造沙棘林提供依据。     

光合和抗氧化能力及相关性分析海桑属红树植物离子积累

以海桑属(Sonneratia Linn. f.)红树植物无瓣海桑(S. apetala Buch.-Ham.)、海桑也S. caseolaris (Linn.) Engl.页、杯萼海桑(S. alba Smith)、卵叶海桑(S. ovata Backer)、拟海桑(S.× gulngai N. C. Duke et B. R. Jackes)和海南海桑(S.× hainanensis W. C. Ko et al.)为研究对象,比较了根际土壤和叶片中离子含量以及叶片光合和叶绿素荧光参数、叶绿素含量、抗氧化酶活性和 O-·2产生速率的差异,并分析了叶片中 Na+和 Cl-含量与部分生理生化指标的相关性。比较结果表明：海桑和拟海桑叶片中 K+含量最高、Na+和 Cl-含量均显著低于其他种类,但它们的根际土壤中Na+和 Cl-含量却较高。供试6种植物中仅海桑叶片对 Cl-的富集系数小于1,各供试种类对 K+、Na+、Cl-、Ca2+和Mg2+的富集系数均大于1;无瓣海桑对离子的富集系数由大至小依次为 Mg2+、K+、Ca2+、Na+、Cl-,其他种类对离子的富集系数由大至小均依次为 K+、Ca2+、Mg2+、Na+、Cl-。无瓣海桑和海桑的 chla、chlb 和总叶绿素含量差异不显著,但均高于其他种类;供试种类的 chla/ chlb 比值均约为3,可能与海桑属植物为阳生植物有关。无瓣海桑的 Pn、Tr 和Gs 均最高,而杯萼海桑的 Pn、Tr 和 Gs 均最低,但6种植物的 Ci 无明显差异。供试种类的 Fv / Fm、qP、ETR 和ΦPSⅡ均无显著差异,仅部分种类间的 NPQ 差异显著。无瓣海桑叶片中 SOD、CAT 和 POD 活性均显著高于其他种类,但O-·2产生速率最低;而卵叶海桑叶片中 O-·2产生速率最高,其 APX 活性也均显著高于其他种类。相关性分析结果表明：供试6种植物叶片中 Na+和 Cl-含量与 Pn、qP、ΦPSⅡ和 ETR 负相关,与 NPQ 及 SOD、CAT、APX 和 POD 活性正相关。其中,Na+含量与 qP、ΦPSⅡ、ETR 和 SOD 活性极显著相关,与 NPQ 和 CAT 活性显著相关;Cl-含量与 SOD 活性极显著相关,与 qP、ΦPSⅡ和 ETR 显著相关。研究结果表明：供试海桑属植物对高盐生境有不同的耐性机制,其中,海桑和拟海桑通过拒吸 Na+和 Cl-抵御盐胁迫的伤害;供试6种植物对海岸潮间带生境的适应性有明显差异,无瓣海桑最适宜在此生境中生长。     

东北虎幼体葡萄糖、四种离子、尿素氮及肌酐含量的测定与分析

用HITACHI 7600-20型全自动生化分析仪对13只东北虎幼体进行了8项生化指标的测定,运用生物统计学分析方法,分别对东北虎幼体雌雄个体间及东北虎幼体与华南虎成体间进行了比较分析.结果表明,东北虎雌雄个体间血清中K 含量呈极差异显著(P<0.01),而葡萄糖、Na 、Ca2 、HCO3-、尿素及肌酐含量并无显著差异;在Cl-、肌酐含量上,雌性东北虎幼体与雌性华南虎有显著差异(P<0.05);雄性东北虎幼体与雄性华南虎在Cl-含量上无显著差异,在葡萄糖及肌酐含量上有极显著差异(P<0.01).     

Investigation of common and specific components of Cl. septicum and Cl. histolyticum toxins

Interrelations between the common and specific components in the toxins of several strains of Cl. septicum and Cl. histolyticum were investigated. The method of tissue culture, which yields more stable results than biological tests on animals, was used. It has been demonstrated that native toxins of Cl. seticum (7 strains) and Cl. histolyticum (7 strains) cause cytotoxic changes in chick embryo fibroblasts. These changes are similar to each other and identical with changes occurring under the effect of concentrated toxins of the mentioned microorganisms. Cross reactions of neutralization with antitoxic and species-specific sera against Cl. septicum and Cl. histolyticum have shown that the strains of Cl. septicum and Cl. histolyticum synthesize toxins with components possessing common antigenic properties. The strains of Cl. histolyticum synthesize a greater amount of components common with Cl. septicum than the strains of Cl. septicum in which the amount of heterologous antigens varies.     

Pleural surface fluorescence measurement of Na+ and Cl- transport across the air space-capillary barrier.

We developed a pleural surface fluorescence method to measure Na(+) and Cl(-) transport in perfused mouse lungs. The air space was filled with aqueous fluid containing membrane-impermeant fluorescent indicators of Cl(-) (lucigenin) or Na(+) (Sodium Green). After instillation of a Cl(-)-free solution into the air space, an increase in perfusate Cl(-) concentration from 0 to 30 mM produced a decrease in surface lucigenin fluorescence (6.5%/min) corresponding to Cl(-) influx of 1.0 mM/min. Cl(-) influx was increased to 2.1 +/- 0.3 mM/min by forskolin, and the increase was inhibited by glibenclamide. cAMP-stimulated Cl(-) influx was decreased by 57% in CFTR null mice. After instillation of a Na(+)-free solution into the air space, an increase in perfusate Na(+) concentration from 0 to 30 mM gave increased Sodium Green fluorescence (Na(+) influx of 1.2 mM/min), which increased approximately fivefold after cAMP agonists. Cl(-) and Na(+) transport were not affected in lungs from mice lacking aquaporins AQP1 or AQP5. Our results establish a pleural surface fluorescence method to measure unidirectional Cl(-) and Na(+) flux in intact lung and provide evidence for cAMP-stimulated transcellular Cl(-) and Na(+) transport.     

Fluorescence measurement of chloride transport in monolayer cultured cells. Mechanisms of chloride transport in fibroblasts.

The methodology has been developed to measure Cl activity and transport in cultured cells grown on a monolayer using the entrapped Cl-sensitive fluorophore 6-methoxy-N-[3-sulfopropyl] quinolinium (SPQ). The method was applied to a renal epithelial cell line, LLC-PKI, and a nonepithelial cell line, Swiss 3T3 fibroblasts. SPQ was nontoxic to cells when present for greater than h in the culture media. To load with SPQ (5 mM), cells were made transiently permeable by exposure to hypotonic buffer (150 mOsm, 4 min). Intracellular fluorescence was monitored continuously by epifluorescence microscopy using low illumination intensity at 360 +/- 5 nm excitation wavelength and photomultiplier detection at greater than 410 nm. Over 60 min at 37 degrees C, there was no photobleaching and less than 10% leakage of SPQ out of cells; intracellular SPQ fluorescence was uniform. SPQ fluorescence was calibrated against intracellular [Cl] using high K solutions containing the ionophores nigericin and tributyltin. The Stern-Volmer constant (Kq) for quenching of intracellular SPQ by Cl was 13 M-1 for fibroblasts and LLC-PKl cells. In the absence of Cl, SPQ lifetime was 26 ns in aqueous solution and 3.7 +/- 0.6 ns in cells, showing that the lower Kq in cells than in free solution (Kq = 118 M-1) was due to SPQ quenching by intracellular anions. To examine Cl transport mechanisms, the time course of intracellular [Cl] was measured in response to rapid Cl addition and removal in the presence of ion or pH gradients. In fibroblasts, three distinct Cl transporting systems were identified: a stilbeneinhibitable Cl/HCO3 exchanger, a furosemide-sensitive Na/K/2Cl cotransporter, and a Ca-regulated Cl conductance. These results establish a direct optical method to measure intracellular [Cl] continuously in cultured cells.     

Development of an accelerated method of determining the antibiotic sensitivity of Cl. perfringens type A]

An express method for determination of antibiotic sensitivity in the strains of Cl. perfringens of type A using Soviet dry nutrient media and antibiotics is proposed. The criteria for estimation of the level of the antibiotic sensitivity of the causative agent of gas gangrene in short periods on the basis of comparison of the data of the antibiotic agar diffusion procedure and the antibiotic MIC were worked out. Twelve antibiotics and 45 collection strains of Cl. perfringens of type A were used in the experiment. The antibiotic agar diffusion method with the use of the nutrient media, microbial load and cultivation conditions developed by the authors is recommended for tentative determination of the antibiotic sensitivity in Cl. perfringens of type A for 4 hours. The use of the agar diffusion method and determination of the antibiotic MIC provided complete estimation of tha antibiotic sensitivity of Cl. perfringens of type A within not more than 24 hours.     

Evaluation of a monoclonal antibody-based immunoassay for detecting type A Clostridium botulinum toxin produced in pure culture and an inoculated model cured meat system

A monoclonal antibody-based amplified enzyme-linked immunosorbent assay (ELISA) method for detecting Clostridium botulinum type A toxin was evaluated for its ability to detect the toxin in the supernatant fluid of pure cultures and after growth from Cl. botulinum spores inoculated into pork slurries. Slurries containing NaCl (1.5-4.5% w/v) and polyphosphate (0.3% w/v) were either unheated or heated, 80 degrees C/5 min + 70 degrees C/2 h, before storage at 15 degrees, 20 degrees or 27 degrees C. The presence of specific toxin was confirmed by mouse bioassay and results compared with those of the amplified ELISA method. A total of 49 strains, 39 Cl. botulinum and 10 Cl. sporogenes (putrefactive anaerobes), and 95 slurry samples were tested. Fourteen of 15 strains of type A Cl. botulinum and 34 of 36 slurry samples containing type A toxin were positive by ELISA. No false positive reactions occurred with Cl. botulinum types B, C, D, E and F, or with the 10 strains of Cl. sporogenes. However, toxin produced by one strain of Cl. botulinum type A (NCTC 2012) was not detected by the amplified ELISA.     

Detection by in vitro amplification of the alpha-toxin (phospholipase C) gene from Clostridium perfringens

A polymerase chain reaction (PCR) with thermostable DNA polymerase from Thermus aquaticus is described for the specific amplification of the phospholipase C (alpha-toxin) gene of Clostridium perfringens. A set of primers selected for their high specificity could detect Cl. perfringens in stools with a detection limit of approximately 5 x 10² bacteria, after bi-amplification. A modified PCR without thermal steps was performed to rapidly amplify, with a yield of 60%, the DNA template. With this PCR method Cl. perfringens alpha-toxin gene could be detected within 2 h. The PCR method detected alpha-toxin positive Cl. perfringens but did not react with phospholipase C-producing Bacillus cereus, Pseudomonas aeruginosa, Cl. sordellii and Cl. bifermentans.
The amplified PCR products were screened through ethidium bromide agarose gel electrophoresis or, in only 1 h, with the PhastSystem (Pharmacia). This PCR satisfies the criteria of specificity, sensitivity and rapidity required for a useful tool in epidemiology and for the diagnosis of the pathogen Cl. perfringens as it may be used directly on stool samples.     

Chlorine‐Enabled Electron Doping in Solution‐Synthesized SnSe Thermoelectric Nanomaterials

An aqueous solution method is developed for the facile synthesis of Cl‐containing SnSe nanoparticles in 10 g quantities per batch. The particle size and Cl concentration of the nanoparticles can be efficiently tuned as a function of reaction duration. Hot pressing produces n‐type Cl‐doped SnSe nanostructured compacts with thermoelectric power factors optimized via control of Cl dopant concentration. This approach, combining an energy‐efficient solution synthesis with hot pressing, provides a simple, rapid, and low‐cost route to high performance n‐type SnSe thermoelectric materials.     

Evaluation of a monoclonal antibody-based immunoassay for detecting type A Clostridium botulinum toxin produced in pure culture and an inoculated model cured meat system

A monoclonal antibody-based amplified enzyme-linked immunosorbent assay (ELISA) method for detecting Clostridium botulinum type A toxin was evaluated for its ability to detect the toxin in the supernatant fluid of pure cultures and after growth from Cl. botulinum spores inoculated into pork slurries. Slurries containing NaCl (1.5–4.5% w/v) and polyphosphate (0.3% w/v) were either unheated or heated, 80°C/5 min + 70°C/2 h, before storage at 15°, 20° or 27°C. The presence of specific toxin was confirmed by mouse bioassay and results compared with those of the amplified ELISA method. A total of 49 strains, 39 Cl. botulinum and 10 Cl. sporogenes (putrefactive anaerobes), aiid 95 slurry samples were tested. Fourteen of 15 strains of type A Cl. botulinum and 34 of 36 slurry samples containing type A toxin were positive by ELISA. No false positive reactions occurred with Cl. botulinum types B, C, D, E and F, or with the 10 strains of Cl. sporogenes. However, toxin produced by one strain of Cl. botulinum type A (NCTC 2012) was not detected by the amplified ELISA.     

Altered expression of claudin family proteins in Alzheimer’s disease and vascular dementia brains

Claudins (Cls) are a multigene family of transmembrane proteins with different tissue distribution, which have an essential role in the formation and sealing capacity of tight junctions (TJs). At the level of the blood–brain barrier (BBB), TJs are the main molecular structures which separate the neuronal milieu from the circulatory space, by a restriction of the paracellular flow of water, ions and larger molecules into the brain. Different studies suggested recently significant BBB alterations in both vascular and degenerative dementia types. In a previous study we found in Alzheimer’s disease (AD) and vascular dementia (VaD) brains an altered expression of occludin, a molecular partner of Cls in the TJs structure. Therefore in this study, using an immunohistochemical approach, we investigated the expression of Cl family proteins (Cl‐2, Cl‐5 and Cl‐11) in frontal cortex of aged control, AD and VaD brains. To estimate the number of Cl‐expressing cells, we applied a random systematic sampling and the unbiased optical fractionator method. We found selected neurons, astrocytes, oligodendrocytes and endothelial cells expressing Cl‐2, Cl‐5 and Cl‐11 at detectable levels in all cases studied. We report a significant increase in ratio of neurons expressing Cl‐2, Cl‐5 and Cl‐11 in both AD and VaD as compared to aged controls. The ratio of astrocytes expressing Cl‐2 and Cl‐11 was significantly higher in AD and VaD as compared to aged controls. The ratio of oligodendrocytes expressing Cl‐11 was significantly higher in AD and the ratio of oligodendrocytes expressing Cl‐2 was significantly higher in VaD as compared to aged controls. Within the cerebral cortex, Cls were selectively expressed by pyramidal neurons, which are the ones responsible for cognitive processes and affected by AD pathology. Our findings suggest a new function of Cl family proteins which might be linked to response to cellular stress.     

Chloride concentration in endosomes measured using a ratioable fluorescent Cl- indicator: evidence for chloride accumulation during acidification.

A novel long wavelength fluorescent Cl(-) indicator was used to test whether endosomal Cl(-) conductance provides the principal electrical shunt to permit endosomal acidification. The green fluorescent Cl(-)-sensitive chromophore 10,10'-bis[3-carboxypropyl]-9,9'-biacridinium dinitrate (BAC) was conjugated to aminodextran together with the red fluorescent Cl(-)-insensitive chromophore tetramethylrhodamine (TMR). BAC fluorescence is pH-insensitive and quenched by Cl(-) with a Stern-Volmer constant of 36 m(-1). Endosomes in J774 and Chinese hamster ovary (CHO) cells were pulse-labeled with BAC-TMR-dextran by fluid-phase endocytosis. Endosomal [Cl(-)] increased over 45 min from 17 to 53 mm in J774 cells and from 28 to 73 mm in CHO cells, during which time endosomal pH decreased from 6.95 to 5.30 (J774) and 6.92 to 5.60 (CHO). The acidification and increased [Cl(-)] were blocked by bafilomycin. Together with ion substitution and buffer capacity measurements, we conclude that Cl(-) transport accounts quantitatively for the electrical shunt during vacuolar acidification. Measurements of relative endosomal volume by a novel ratio imaging method involving fluorescence self-quenching indicated a 2.5-fold increase in volume during early acidification and Cl(-) accumulation, which was blocked by bafilomycin. These experiments provide the first direct measurement of endosomal [Cl(-)] and indicate that endosomal acidification is accompanied by significant Cl(-) entry and volume increase.