Protein aggregation as primary and characteristic cell reaction to various stresses

Ehrlich carcinoma and EL-4 thymoma ascites cells were subjected in vitro to heat shock, ATP depletion, oxidative stress, Ca²⁺ overlading and iodoacetamide treatment. After the transient stresses, Triton (X-100)-insoluble TIS) fractions were isolated from the cells and analysed by electrophoresis and immunoblotting. All stresses used caused rapid aggregation of cell proteins. This was manifested in a signficant rise in protein content in the TIS fractions. The protein increase was mostly due to and increase in the insolubility of actin, 57 kDa protein of intermediate filaments, 70 kDa heat shock protein (HSP 70), and some specific proteins whose insolubilization was a characteristic sign for each type of cell injury. Different survival rates in the cell lines after either stress corrlated well with differences in their TIS protein accretion. Possible mechanisms for stress-induced protein aggregation and its relationship with cell viability are suggested.     

热休克对粗糙脉孢菌核酸内外切酶的影响

将快速生长的粗糙脉孢菌Ｎｅｕｒｏｓｐｏｒａｃｒａｓｓａ培养液从３０℃升温至４５℃，热休克处理２小时，７５％有活性的核酸内外切酶ｅｎｄｏ－ｅｘｏｎｕｌｅａｓｅ（此酶为ＤＮＡ修复酶）从核及液泡中释放到胞质并以无活性的形式储存起来：用放线菌酮处理，不能阻止此酶的释放，但能使灭活降低；致使胞质的酶活性提高５－１５倍。热休克菌体核酸内外切酶的灭活是由于某种抑制物质的作用，后者来自酶前体的自身降解。作者推测由热休克诱导了一种蛋白酶，此酶使核酸内外切酶前体水解为抑制物质。     

颗粒分布对金属基复合材料激光热冲击破坏的影响

从宏观和微观两方面研究了ＳｉＣ颗粒分布状况对陶瓷颗粒增强金属基复合材料激光热冲击破坏的影响,发现微裂纹形成是由基体内的孔洞和颗粒与基体脱胶所引起,而陶瓷颗粒分布不均匀对裂纹的扩展机理有重要影响,它大大降低了复合材料的宏观力学性能     

A proteomic analysis of bacterial strain<Emphasis Type="Italic">Sinorhizobium fredii</Emphasis> RT19 subjected to salt shock

Sinorhizobium fredii RT19, a strain of freeliving bacteria, was subjected to salt shock and its protein expression profiles were analyzed by differential display proteome approaches. The results of separation by two-dimensional polyacrylamide gel electrophoresis (2D PAGE) showed that the number of resolved proteins was 481, 465 and 424, corresponding to salt-free control, 5 and 50 min 1 mol/L salt treatment, respectively. Among the resolved proteins, 82 in total had altered expression in response to salt-shock stress. 26 out of the 82 proteins were induced and 23 were completely inhibited, while 12 were up-regulated and 21 down-regulated in response to salt shock. In addition, the appearance of differentially displayed proteins responding to different salt shock periods is also reported. The identity of the 26 induced proteins was revealed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) followed by database searching. Among them, 20 were assigned to proteins with known functions. Their roles in response to salt shock stress are discussed.     

Identification of swelling-activated Cl<Superscript>-</Superscript> current in rabbit cardiac Purkinje cells

The presence and functional role of the swelling-activated Cl^- current (I_Cl(swell)) in rabbit cardiac Purkinje cells was examined using patch-clamp methodology. Extracellular hypotonicity (210 or 135 mOsm) activated an outwardly rectifying, time-independent current with a reversal potential close to the calculated Cl^- equilibrium potential (E_Cl). The magnitude of this current was related to tonicity of the superfusate. The current was blocked by 0.5 mM 4,4-diisothiocyanostilbene-2,2-disulfonic acid (DIDS). These features are comparable to those of I_Cl(swell) found in sinoatrial nodal, atrial, and ventricular myocytes. I_Cl(swell) activation at 210 and 135 mOsm depolarized the resting membrane potential with 6 and 10 mV and shortened the action potential by 18 and 33%, respectively. DIDS partially reversed I_Cl(swell)-induced action potential changes. We conclude that I_Cl(swell) is present in Purkinje cells and its activation leads to action potential shortening and resting membrane potential depolarization, both of which can promote the development of reentrant arrhythmias.Received 20 January 2004; received after revision 17 February 2004; accepted 25 February 2004     

FK506 sensitizes mammalian cells to high osmolarity by modulating p38 MAP kinase activation

The immunosuppressants tacrolimus (FK506) and cyclosporin A (CsA) have increased the survival rates in organ transplantation. Both drugs inhibit the protein phosphatase calcineurin (CaN) in activated T cells, exhibiting similar side-effects. Diabetes is observed more often in FK506 than CsA therapy, probably due to inhibition of new molecular targets other than CaN. We studied FK506 toxicity in mammalian cells. FK506, but not CsA, regulated p38 activation by osmotic stress, and decreased viability in osmostressed cells. In addition, FK506 treatment strongly increased the phosphorylation of the eukaryotic initiation factor-2a (eIF-2a) subunit. eIF-2a phosphorylation, p38 inhibition and cell lethality were relieved by addition of excess amino acids to the medium, suggesting that amino acid availability mediated FK506 toxicity. Therefore, these FK506-dependent responses could be relevant to the non-therapeutic effects of FK506 therapy.Received 16 October 2003; received after revision 8 January 2004; accepted 14 January 2004     

海藻糖对渗透胁迫下油菜种子萌发和幼苗生长的影响

25％PEG-6000渗透胁迫显著抑制油菜种子萌发和幼苗生长．使种子萌发推迟，发芽率降低22.67％，发芽指数降低112．8％，活力指数降低10．4倍，单株鲜质量、地上与地下部分鲜质量、地上和地下部分长度分别降低4．2，4．0g，4．82，4．19和1.42倍；25mmol／1。海藻糖（TH）处理能显著促进渗透胁迫下油菜种子萌发和幼苗生长．使种子发芽率提高14．67％，发芽指数提高33．12％，活力指数提高175．40％，单株鲜质量提高101．69％，地上、地下部分鲜质量和地上、地下部分长度分别提高62．38％，335．29％，25％和11．11％．     

复合式氧化沟处理城市废水运行条件的控制研究

采用复合式氧化沟工艺处理城市污水,并对系统的运行条件进行了探讨.实验结果表明:当水停留时间HRT≥30 h,COD、BOD5和氨氮的去除率保持在96%、97%和93%以上,当HRT分别为20 h和10 h时,COD、BOD5和氨氮分别为90%、94%、87%和86%、92%、83%;重点考察在HRT为20 h时,COD的浓度范围从125.3~492 mg/L,氨氮的浓度范围为20~32.91 mg/L,但是它们的去除率维持在90%以上,系统对污染物有更高更稳定的去除效果,而且系统对冲击负荷也有很强的适应性.