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Autophagy, apoptosis and organelle features during cell exposure to cadmium. 镉对细胞自噬、凋亡和细胞器的影响。
C. S. Vergílio, E. D. de Melo
Cadmium (Cd) induces several effects in different tissues, but our knowledge of the toxic effects on organelles is insufficient. To observe the progression of Cd effects on organelle structure and function, HuH-7 cells (human hepatic carcinoma cell line) were exposed to CdCl2 in increasing concentrations (1 microM - 20 microM) and exposure times (2 h - 24 h). During Cd treatment, the cells exhibited a progressive decrease in viability that was both time- and dose-dependent. Cd treated cells displayed progressive morphological changes that included cytoplasm retraction and nuclear condensation preceding a total loss of cell adhesion. Treatment with 10 microM for 12 h led to irreversible damages. Before these drastic and irreparable damages, treated cells (5 microM for 12 h) presented a progressive loss of mitochondrial function and cytoplasm acidification as well as dysfunction and disorganization of microfilaments and endoplasmic reticulum. These damages led to the induction of apoptotic events and an increase in autophagic bodies in the cytoplasm. These results revealed that Cd affects multiple intra-cellular targets that induce alterations in the mitochondria, cytoskeleton, endoplasmic reticulum and acidic compartments, ultimately culminating in cell death via apoptotic and autophagic pathways.
镉(Cd)在不同的组织中引起几种作用,但我们对其对细胞器的毒性作用的认识还不够。为了观察Cd对细胞器结构和功能影响的进展,我们将HuH-7细胞(人肝癌细胞系)暴露在CdCl2浓度(1微米- 20微米)和暴露时间(2小时- 24小时)增加的环境中。在Cd处理过程中,细胞的活力表现出时间和剂量依赖性的逐渐下降。Cd处理的细胞表现出进行性的形态学改变,包括细胞质收缩和细胞核凝聚,直至完全丧失细胞粘附。10 μ m处理12 h可导致不可逆损伤。在这些剧烈和不可修复的损伤之前,处理细胞(5微米12小时)表现出线粒体功能的进行性丧失和细胞质酸化,以及微丝和内质网的功能障碍和解体。这些损伤导致凋亡事件的诱导和细胞质中自噬小体的增加。这些结果表明,Cd影响多种细胞内靶点,诱导线粒体、细胞骨架、内质网和酸性室的改变,最终通过凋亡和自噬途径导致细胞死亡。
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引用次数: 12
Meta-analysis of the cell cycle related C12orf48. 细胞周期相关C12orf48的meta分析。
L. Varışlı
The cell cycle is a conserved process from yeast to mammals and focuses on mechanisms that regulate the timing and frequency of DNA replication and cell division. The temporal and spatial expression of the genes is tightly regulated to ensure accurate replication and transmission of DNA to daughter cells during the cycle. Although the genes involved in interphase are well studied, most of the genes which are involved in mitotic events still remain unidentified. Since, the discovery of mitosis related genes is still incomplete, we performed a co-expression and gene ontology analysis for revealing novel mitosis regulated genes. In this study, we showed that C12orf48 is co-expressed with well-known mitotic genes. Moreover, it is also co-expressed with the genes that have roles in interphase such as DNA replication. Furthermore, our results showed that C12orf48 is also differentially expressed in various cancers. Therefore, the results presented in this study suggest that C12orf48 may be an important molecule for both interphase and mitosis. Since, the molecules involved in these mechanisms are crucial for proliferation as well as in carcinogenesis, C12orf48 should be considered as a novel cell cycle and carcinogenesis related gene.
细胞周期是一个保守的过程,从酵母到哺乳动物,重点是调节DNA复制和细胞分裂的时间和频率的机制。基因的时空表达受到严格调控,以确保DNA在周期中准确复制和传递到子细胞。虽然参与间期的基因已被研究得很好,但大多数参与有丝分裂事件的基因仍未被识别。由于有丝分裂相关基因的发现仍然不完整,我们进行了共表达和基因本体分析,以揭示新的有丝分裂调节基因。在这项研究中,我们发现C12orf48与众所周知的有丝分裂基因共表达。此外,它还与在间期(如DNA复制)中起作用的基因共表达。此外,我们的研究结果表明,C12orf48在各种癌症中也存在差异表达。因此,本研究结果提示C12orf48可能是间期和有丝分裂的重要分子。由于参与这些机制的分子在增殖和癌变中都是至关重要的,因此C12orf48应该被认为是一个新的细胞周期和癌变相关基因。
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引用次数: 7
High passage numbers induce resistance to apoptosis in C2C12 muscle cells. 高传代数诱导C2C12肌肉细胞对凋亡的抗性。
Lucía Pronsato, Anabela La Colla, A. Ronda, L. Milanesi, R. Boland, A. Vasconsuelo
Cell lines with high passage numbers exhibit alterations in cell morphology and functions. In the present work, C2C12 skeletal muscle cells with either low (< 20) or high (> 60) passage numbers (identified as 1-C2C12 or h-C2C12, respectively) were used to investigate the apoptotic response to H2O2 as a function of culture age h-C2C12. We found that older cultures (h-C2C12 group) were depleted of mitochondrial DNA (mtDNA). When we analyzed the behavior of Bad, Bax, caspase-3 and mitochondrial transmembrane potential, we observed that cells in the h-C2C12 group were resistant to H2O2 induction of apoptosis. We propose serially cultured C2Cl2 cells as a refractory model to H2O2-induced apoptosis. In addition, the data obtained in this work suggest that mtDNA is required for apoptotic cell death in skeletal muscle C2C12 cells.
传代数高的细胞系表现出细胞形态和功能的改变。本研究采用低传代数(< 20)或高传代数(bbb60)的C2C12骨骼肌细胞(分别鉴定为1-C2C12或h-C2C12),研究H2O2对细胞凋亡的影响与h-C2C12培养年龄的关系。我们发现较老的培养(h-C2C12组)线粒体DNA (mtDNA)被耗尽。当我们分析Bad、Bax、caspase-3和线粒体跨膜电位的行为时,我们观察到h-C2C12组细胞对H2O2诱导的细胞凋亡具有抗性。我们提出连续培养的C2Cl2细胞作为h2o2诱导的细胞凋亡的难治性模型。此外,本工作获得的数据表明,mtDNA是骨骼肌C2C12细胞凋亡细胞死亡所必需的。
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引用次数: 14
A likely role for a novel PH-domain containing protein, PEPP2, in connecting membrane and cytoskeleton. 一个新的含有ph结构域的蛋白PEPP2在连接膜和细胞骨架中的可能作用。
Yi Zou, Wen-Sheng Zhong
PH domains (pleckstrin homology) are well known to bind membrane phosphoinositides with different specificities and direct PH domain-containing proteins to discrete subcellular apartments with assistances of alternative binding partners. PH domain-containing proteins are found to be involved in a wide range of cellular events, including signalling, cytoskeleton rearrangement and vesicular trafficking. Here we showed that a novel PH domain-containing protein, PEPP2, displayed moderate phosphoinositide binding specificity. Full length PEPP2 associated with both plasma membrane and microtubules. The membrane-associated PEPP2 nucleated at cell-cell contacts and the leading edge of migrating cells. Overexpression of PEPP2 increased membrane microviscosity, indicating a potential role of PEPP2 in regulating function of membrane and microtubules.
众所周知,PH结构域(pleckstrin同源性)可以结合不同特异性的膜磷酸肌苷,并在其他结合伙伴的帮助下将含PH结构域的蛋白质直接结合到离散的亚细胞公寓。含有PH结构域的蛋白质被发现参与了广泛的细胞事件,包括信号传导、细胞骨架重排和囊泡运输。在这里,我们发现了一种新的含有PH结构域的蛋白PEPP2,显示出中等的磷酸肌苷结合特异性。全长PEPP2与质膜和微管相关。膜相关的PEPP2在细胞间接触处和迁移细胞的前缘成核。PEPP2的过表达增加了膜的微粘度,表明PEPP2可能在调节膜和微管的功能中起潜在的作用。
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引用次数: 12
Reproductive performance of the Mesa silverside (Chirostoma jordani Woolman, 1894) under natural and controlled photoperiods. 在自然和受控光周期下银台菜(Chirostoma jordani Woolman, 1894)的繁殖性能。
J. L. Arredondo-Figueroa, L. Núñez-García, Paloma Adriana Heredia-Guzmán, J. Ponce‐Palafox
Chirostoma jordani is a native annual species inhabiting lacustrine waters of the Central Mexico Plateau. It is widely distributed and is currently facing high environmental pressures. Five experiments were performed to study the reproductive performance of this species. Four of the experiments were conducted in 270-L indoor recirculation tanks. Two males and one female at the first stage of reproduction were included in each test. A photoperiod of 14 light hours and 10 dark hours was used. In a fifth experiment, 10 females and 15 males were kept in an outdoor 3,000-L recirculation tank under natural photoperiod. The number of spawns, fertilised eggs and 30-day-old juveniles were counted and the survival rate was calculated. The results indicated significant differences (P < 0.05) between treatments. Higher spawn numbers and greater egg production were observed under controlled photoperiod, and higher numbers of juveniles and a higher survival rate were observed under natural photoperiod. The trials exhibited different patterns of egg production during the experiment. The egg production in the natural-photoperiod trials followed a polynomial curve model. In contrast, the trials under the controlled photoperiod showed an irregular pattern of increases and decreases in egg production.
约旦Chirostoma jordani是栖息在墨西哥中部高原湖泊水域的本地一年生物种。它分布广泛,目前面临着很大的环境压力。通过5项试验研究了该物种的繁殖性能。其中4个实验在270-L的室内循环水箱中进行。每个试验包括两个雄性和一个雌性在繁殖的第一阶段。光周期为14光小时和10暗小时。试验5:在自然光周期条件下,将10尾雌鱼和15尾雄鱼饲养在3000 l的室外循环池中。计数产卵数、受精卵数和30日龄幼鱼数,计算成活率。两组间差异有统计学意义(P < 0.05)。控制光周期下的产卵量和产蛋量较高,自然光周期下的幼鱼数量和成活率较高。在试验过程中,试验显示出不同的产蛋模式。自然光周期试验的产蛋量服从多项式曲线模型。相反,控制光周期的试验显示出不规则的产蛋量增减规律。
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引用次数: 1
Effects of rotational culture on morphology, nitric oxide production and cell cycle of endothelial cells. 旋转培养对内皮细胞形态、一氧化氮生成和细胞周期的影响。
Chao-jun Tang, Xue Wu, L. Ye, Xiang Xie, Guixue Wang
Devices for the rotational culture of cells and the study of biological reactions have been widely applied in tissue engineering. However, there are few reports exploring the effects of rotational culture on cell morphology, nitric oxide (NO) production, and cell cycle of the endothelial cells from human umbilical vein on the stent surface. This study focuses on these parameters after the cells are seeded on the stents. Results showed that covering of stents by endothelial cells was improved by rotational culture. NO production decreased within 24 h in both rotational and static culture groups. In addition, rotational culture significantly increased NO production by 37.9% at 36 h and 28.9% at 48 h compared with static culture. Flow cytometry showed that the cell cycle was not obviously influenced by rotational culture. Results indicate that rotational culture may be helpful for preparation of cell-seeded vascular grafts and intravascular stents, which are expected to be the most frequently implanted materials in the future.
细胞旋转培养和生物反应研究装置在组织工程中得到了广泛的应用。然而,很少有报道探讨旋转培养对支架表面人脐静脉内皮细胞形态、一氧化氮(NO)生成和细胞周期的影响。本研究的重点是细胞植入支架后的这些参数。结果表明,旋转培养可改善内皮细胞对支架的覆盖。旋转培养组和静态培养组24 h内NO产量均下降。此外,与静态培养相比,轮作培养36 h和48 h的NO产量分别显著提高了37.9%和28.9%。流式细胞术显示,旋转培养对细胞周期影响不明显。结果表明,旋转培养可能有助于制备细胞种子血管移植物和血管内支架,这有望成为未来最常用的植入材料。
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引用次数: 2
Structural analysis of flagellar axonemes from inner arm dynein knockdown strains of Trypanosoma brucei. 布鲁氏锥虫内臂动力蛋白敲除株鞭毛轴突的结构分析。
Randi Zukas, Alex J. Chang, Marian Rice, Amy Springer
Trypanosoma brucei is a protozoan flagellate that causes African sleeping sickness. Flagellar function in this organism is critical for life cycle progression and pathogenesis, however the regulation of flagellar motility is not well understood. The flagellar axoneme produces a complex beat through the precisely coordinated firing of many proteins, including multiple dynein motors. These motors are found in the inner arm and outer arm complexes. We are studying one of the inner arm dynein motors in the T. brucei flagellum: dynein-f. RNAi knockdown of genes for two components of dynein-f: DNAH10, the alpha heavy chain, and IC138, an intermediate chain, cause severe motility defects including immotility. To determine if motility defects result from structural disruption of the axoneme, we used two different flagellar preparations to carefully examine axoneme structure in these strains using transmission electron microscopy (TEM). Our analysis showed that inner arm dynein size, axoneme structural integrity and fixed central pair orientation are not significantly different in either knockdown culture when compared to control cultures. These results support the idea that immotility in knockdowns affecting DNAH10 or IC138 results from loss of dynein-f function rather than from obvious structural defects in the axoneme.
布氏锥虫是一种能引起非洲昏睡病的鞭毛原虫。鞭毛的功能在这种生物的生命周期进展和发病机制中至关重要,然而鞭毛运动的调控尚不清楚。鞭毛轴突通过许多蛋白质(包括多个动力蛋白马达)的精确协调发射产生复杂的节拍。这些马达存在于内臂和外臂复合体中。我们正在研究布鲁氏体毛内臂动力蛋白马达之一:动力蛋白-f。RNAi敲低dynein-f的两个组成部分基因:DNAH10 (α重链)和IC138(中间链),会导致包括不运动在内的严重运动缺陷。为了确定运动缺陷是否由轴突结构破坏引起,我们使用了两种不同的鞭毛制剂,用透射电子显微镜(TEM)仔细检查了这些菌株的轴突结构。我们的分析表明,与对照培养相比,两种敲除培养中手臂内动力蛋白的大小、轴素结构完整性和固定的中心对取向没有显著差异。这些结果支持了这样一种观点,即影响DNAH10或IC138的敲低不动是由于动力蛋白f功能的丧失,而不是由于轴突中明显的结构缺陷。
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引用次数: 14
Knockdown of apoptosis-inducing factor disrupts function of respiratory complex I. 凋亡诱导因子的下调可破坏呼吸复合体I的功能。
M. Varecha, D. Páclová, J. Procházková, P. Matula, D. Cmarko, M. Kozubek
Recent findings suggest that apoptotic protein apoptosis-inducing factor (AIF) may also play an important non-apoptotic function inside mitochondria. AIF was proposed to be an important component of respiratory chain complex I that is the major producer of superoxide radical. The possible role of AIF is still controversial. Superoxide production could be used as a valuable measure of complex I function, because the majority of superoxide is produced there. Therefore, we employed superoxide-specific mitochondrial fluorescence dye for detection of superoxide production. We studied an impact of AIF knockdown on function of mitochondrial complex I by analyzing superoxide production in selected cell lines. Our results show that tumoral telomerase-positive (TP) AIF knockdown cell lines display significant increase in superoxide production in comparison to control cells, while a non-tumoral cell line and tumoral telomerase-negative cell lines with alternative lengthening of telomeres (ALT) show a decrease in superoxide production. According to these results, we can conclude that AIF knockdown disrupts function of complex I and therefore increases the superoxide production in mitochondria. The distinct effect of AIF depletion in various cell lines could result from recently discovered activity of telomerase in mitochondria of TP cancer cells, but this hypothesis needs further investigation.
最近的研究表明,凋亡蛋白凋亡诱导因子(AIF)在线粒体内也可能发挥重要的非凋亡功能。AIF被认为是呼吸链复合体I的重要组成部分,而呼吸链复合体I是超氧自由基的主要产生者。AIF可能扮演的角色仍存在争议。超氧化物的产生可以作为复合体I功能的一个有价值的测量,因为大部分的超氧化物是在那里产生的。因此,我们采用超氧化物特异性线粒体荧光染料检测超氧化物的产生。我们通过分析选定细胞系的超氧化物产生,研究了AIF敲除对线粒体复合体I功能的影响。我们的研究结果表明,与对照细胞相比,肿瘤端粒酶阳性(TP) AIF敲低细胞系的超氧化物产量显著增加,而非肿瘤细胞系和端粒选择性延长(ALT)的肿瘤端粒酶阴性细胞系的超氧化物产量减少。根据这些结果,我们可以得出结论,AIF敲低会破坏复合物I的功能,从而增加线粒体中超氧化物的产生。最近发现的TP癌细胞线粒体端粒酶活性可能是AIF耗竭对多种细胞系产生不同影响的原因,但这一假设有待进一步研究。
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引用次数: 2
Pollen viability of Polygala paniculata L. (Polygalaceae) using different staining methods. 不同染色方法对茯苓花粉活力的影响。
V. Frescura, H. D. Laughinghouse, Thaís Scotti Do Canto-Dorow, S. Tedesco
Polygala paniculata L. is a medicinal plant that grows in the Brazilian Atlantic coast, known as 'barba-de-São-João', 'barba-de-bode', 'vassourinha branca', and 'mimosa'. In this study, pollen viability was estimated by three different staining methods: 2% acetic orcein, 2% acetic carmine, and Alexander's stain. The young inflorescences of twenty accessions were collected and fixed in a solution of ethanol: acetic acid (3:1) for 24 hours, then stored in ethanol 70% under refrigeration. Six slides per plant, two for each stain, were prepared by squashing, and 300 pollen grains per slide were analyzed. Pollen viability was high (> 70%) for most accessions of P. paniculata using the Alexander's stain, which proved the most adequate method to estimate pollen viability.
paniculata L.是一种药用植物,生长在巴西大西洋沿岸,被称为“barba-de- s o- jo o”,“barba-de-bode”,“vassourinha branca”和“含羞草”。在本研究中,花粉活力通过三种不同的染色方法来估计:2%醋酸皂素,2%醋酸胭脂红和亚历山大染色。收集20个材料的幼花序,在乙醇:乙酸(3:1)的溶液中固定24小时,然后在70%的乙醇中冷藏保存。每株6张载玻片,每张染色2张,每张载玻片分析300粒花粉。Alexander’s染色法测定的多数参试品花粉活力均较高(> 70%),是测定参试品花粉活力的最佳方法。
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引用次数: 23
Age related interaction of dopamine and serotonin synthesis in striatal synaptosomes. 纹状体突触体中多巴胺和血清素合成的年龄相关性。
M. Messripour, A. Mesripour
Tyrosine hydroxylase and tryptophan hydroxylase are key rate limiting enzymes in the biosynthesis of dopamine and serotonin, respectively. Since both enzymes are active in striatum, and affected by age, this study was undertaken to investigate interaction between dopamine and serotonin synthesis in brain striatal synaptosomes of aging rat. Male Wistar rats (3 and 30 month old) were killed by decapitation and brain striatal synaptosomes were prepared by discontinuous Ficoll/sucrose gradient technique. Synaptosomes were incubated in the presence of added pargiline (monoamineoxidase inhibitor), dopamine or serotonin synthesized during 25 min was measured by HPLC, employing electrochemical detection. Dopamine synthesis in synaptosomes prepared from young animals was markedly inhibited by addition of 5 microM serotonin concentrations (30%) and increasing serotonin concentrations up to 50 microM caused only a smaller additional inhibition. Dopamine synthesis in synaptosomes obtained from old rats was significantly lower than that of youg animals and addition of serotonin concentrations up to 50 microM had little effect on these preparations. In case of serotonin synthesis, exogenously added 5 microM dopamine inhibited serotonin synthesis in the synaptosomes of both ages by about 40%, whereas with higher concentration of dopamine (10-50 microM) the rate of inhibition was highly pronounced in old rats as compared to that of young animals. It is concluded that dopamine and serotonin interaction may be significant, and that these should be considered in long-term treatments of Parkinson's disease with L-DOPA.
酪氨酸羟化酶和色氨酸羟化酶分别是生物合成多巴胺和血清素的关键限速酶。由于这两种酶在纹状体中都很活跃,且受年龄的影响,本研究探讨了衰老大鼠脑纹状体突触体中多巴胺和血清素合成的相互作用。采用断头法处死3、30月龄雄性Wistar大鼠,采用不连续Ficoll/蔗糖梯度法制备纹状体突触体。突触体在添加pargiline(单胺氧化酶抑制剂)的条件下孵育,用HPLC测定25 min合成的多巴胺或5 -羟色胺,电化学检测。在幼龄动物制备的突触体中,添加5 μ m血清素浓度(30%)可显著抑制多巴胺的合成,而将血清素浓度增加到50 μ m时,只会产生较小的额外抑制作用。老年大鼠突触体中多巴胺的合成明显低于幼鼠,血清素浓度达到50微米时对这些制剂的影响不大。在5 -羟色胺合成方面,外源性添加5 μ m多巴胺可抑制两种年龄大鼠突触体中5 -羟色胺的合成,其抑制率约为40%,而当多巴胺浓度较高(10-50 μ m)时,老年大鼠的抑制率明显高于幼鼠。由此得出结论,多巴胺和血清素的相互作用可能是显著的,在长期使用左旋多巴治疗帕金森病时应考虑到这一点。
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引用次数: 12
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Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al
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