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Protein Expression within the Human Renal Cortex and Renal Cell Carcinoma: The Implication of Cold Ischemia 人肾皮质蛋白表达与肾细胞癌:冷缺血的意义
Pub Date : 2007-08-01 DOI: 10.1089/CPT.2007.0001
H. Gudmundsdóttir, Fjola Haraldsdottir, Ásdís Baldursdóttir, R. Barkardottir, S. Hardarson, V. Gudnason, T. Gudbjartsson, G. Einarsson, E. Jonsson, V. Pétursdóttir
Cold ischemia of tissue during tissue treatment may influence protein expression, but has not been well studied. Better understanding of this is fundamental prior to using stored fresh-frozen tissue where the time from organ harvest until tissue collection and storage is most often not documented. We collected samples from normal renal cortex and cancerous tissues at serial time points for up to 60 min from three nephrectomized individuals with newly diagnosed clear cell renal cell carcinoma (RCC). Samples were processed onto protein chips and identified using surface-enhanced laser desorption/ionization-time of flight mass spectrometry (SELDI). The number and size of proteins expressed at separate sites within homogenous tissue sections were comparable. Cold ischemia time neither affected the number nor the size of proteins expressed. While the quantity of most proteins was similar between separate sites and unaffected by cold ischemia time, we noted variation in the quantity of some proteins compared to...
组织处理过程中组织冷缺血可能影响蛋白表达,但尚未得到很好的研究。在使用储存的新鲜冷冻组织之前,更好地了解这一点是至关重要的,因为从器官采集到组织收集和储存的时间通常没有记录。我们在连续时间点收集了三名新诊断为透明细胞肾细胞癌(RCC)的肾切除术患者的正常肾皮质和癌组织样本,时间长达60分钟。样品被处理到蛋白质芯片上,并使用表面增强激光解吸/电离飞行时间质谱(SELDI)进行鉴定。在均质组织切片中,在不同位点表达的蛋白的数量和大小具有可比性。冷缺血时间对表达蛋白的数量和大小均无影响。虽然大多数蛋白的数量在不同的位点之间是相似的,并且不受冷缺血时间的影响,但我们注意到一些蛋白的数量与…
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引用次数: 1
Water Sorption and Glass Transition Behavior of Polyalditol (PD30), a New Nonreactive Malto-Oligosaccharide Cryo- and Drying-Protectant 新型非反应性麦芽寡糖冷冻干燥保护剂聚醛醇(PD30)的吸水和玻璃化转变行为
Pub Date : 2007-08-01 DOI: 10.1089/CPT.2007.0503
Wendell Q. Sun
Polyalditols are a new family of nonreactive polyols whose reducing power is reduced to <1% dextrose equivalent (DE) by catalytic hydrogenation of malto-oligosaccharides. This work investigated water sorption property and glass transition behavior of PD30, a polyalditol with an average molecular weight of ∼1000 Daltons and a DE value of <0.5. Properties of PD30 are compared with those of maltodextrin M180 (precursor of PD30), which has a DE of 16.5 to 19.5 and is used to make PD30. Water sorption properties were investigated by isothermal sorption and desorption methods. Sorption study showed that PD30 adsorbs less water than M180 at 22°C at relative humidity (RH) below 45%, but adsorbs more water at higher RH. The analysis with Guggenheim-Anderson-de Boer (GAB) model estimates the same monolayer hydration (∼0.05 g/g) for both M180 and PD30, but PD30 could adsorb more weakly bound water. Thermal desorption study showed that PD30 binds significantly less water than M180 at temperatures above 45°C. Glass tr...
聚糖醇是一类新的非反应性多元醇,其还原能力通过麦芽糖低聚糖的催化加氢而降低到<1%葡萄糖当量(DE)。本工作研究了PD30的吸水性能和玻璃化转变行为,PD30是一种平均分子量为~ 1000道尔顿,DE值<0.5的聚醛醇。将PD30的性能与用于制备PD30的DE为16.5 ~ 19.5的麦芽糖糊精M180 (PD30的前体)的性能进行了比较。采用等温吸附和解吸法研究了水的吸附性能。吸附研究表明,在22℃条件下,相对湿度(RH)低于45%时,PD30吸附的水分比M180少,而在较高的相对湿度下,PD30吸附的水分更多。采用Guggenheim-Anderson-de Boer (GAB)模型进行分析,估计M180和PD30的单层水合作用相同(~ 0.05 g/g),但PD30可以吸附更弱的结合水。热解吸研究表明,在45°C以上的温度下,PD30与M180的结合力明显低于M180。玻璃tr……
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引用次数: 1
Genomic Diversity Research and the Role of Biorepositories 基因组多样性研究和生物库的作用
Pub Date : 2007-08-01 DOI: 10.1089/CPT.2007.9993
Robert H. Hanner, T. R. Gregory
Biodiversity repositories underpin the future of research in the life sciences and biotechnology. However, they represent an extremely heterogeneous assemblage of collections that are lacking a comprehensive index of available resources. A set of “best practices” for biospecimen characterization is proposed for repository biomaterials involving standardized species-level molecular genotyping (DNA barcoding) and the quantification of nuclear DNA content (genome size). This approach has implications for upstream sample collection and preservation methods, as well as downstream implications for highlighting biorepository specimens available for genetic and genomic research. The broad application of the approach here proposed will raise the profile of participating biodiversity repositories, facilitate the compilation of validated reference sequences for molecular species recognition, and drive a deeper understanding of the evolution of the genome.
生物多样性资源库是未来生命科学和生物技术研究的基础。然而,它们代表了一个极其异构的集合,缺乏可用资源的综合索引。提出了一套生物标本鉴定的“最佳实践”,涉及标准化物种水平的分子基因分型(DNA条形码)和核DNA含量的量化(基因组大小)。这种方法对上游的样本收集和保存方法有影响,对下游的遗传和基因组研究的生物库标本也有影响。本文提出的方法的广泛应用将提高参与生物多样性库的形象,促进分子物种识别的有效参考序列的编制,并推动对基因组进化的更深入理解。
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引用次数: 29
The Continuing Crisis in Biobanking 生物银行的持续危机
Pub Date : 2007-04-23 DOI: 10.1089/CPT.2007.9992
J. Baust
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引用次数: 0
Comparison of a Selection of Rapid Automated DNA and RNA Extraction Technologies for Detection of Somatic or Constitutional Gene Abnormalities in Cancer Diagnosis 快速自动DNA和RNA提取技术在癌症诊断中检测体细胞或体质基因异常的比较
Pub Date : 2007-04-23 DOI: 10.1089/CPT.2006.9989
F. Lassailly, M. Mozziconacci, T. Noguchi, Didier Bechlian, J. Adélaı̈de, F. Fina, P. Martin, H. Sobol, L. Xerri, D. Birnbaum, C. Chabannon
Molecular analyses of large numbers of patient samples are increasingly used for diagnostic applications as well as for understanding cancer biology. Their accuracy depends on the quality and quantity of nucleic acids extracted from human cells or tissues. To optimize these preanalytical steps, we evaluated several automated technologies for nucleic acid purification from clinical samples. Three automated platforms were compared. DNA was extracted from peripheral blood leukocytes from five normal individuals, and its quality was assessed by D-HPLC and sequencing after PCR. Clinical samples from acute leukemia patients were used for automated RNA extractions; results were compared to our standard manual technique. RNA qualification was done using capillary gel electrophoresis and analysis of the Abelson gene transcript by real-time PCR. One robot produced higher total output for both DNA and RNA. While the quality of DNAs obtained from the three workstations allowed implementation of their analysis for det...
大量患者样本的分子分析越来越多地用于诊断应用以及了解癌症生物学。它们的准确性取决于从人体细胞或组织中提取的核酸的质量和数量。为了优化这些分析前步骤,我们评估了几种用于临床样品核酸纯化的自动化技术。对三种自动化平台进行了比较。从5名正常人的外周血白细胞中提取DNA,采用D-HPLC法和PCR测序法评价其质量。采用急性白血病患者临床标本进行RNA自动提取;结果与我们的标准手工技术进行了比较。用毛细管凝胶电泳鉴定RNA,用实时荧光定量PCR分析Abelson基因转录物。一个机器人对DNA和RNA的总产出更高。虽然从三个工作站获得的dna质量允许实施他们的分析…
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引用次数: 2
Effect of Slow Freezing Versus Vitrification on the Recovery of Mouse Embryonic Stem Cells 慢速冷冻与玻璃化冷冻对小鼠胚胎干细胞恢复的影响
Pub Date : 2007-04-23 DOI: 10.1089/CPT.2006.9990
H. Miszta-Lane, P. Gill, M. Mirbolooki, J. Lakey
The purpose of this study was to cryopreserve mouse embryonic stem (ES) cells R1 line and determine cell viability, morphology, the number of colonies, and the Alkaline Phosphatase (AP) activity. In addition, the expression of transcription factors such as the stage-specific antigen (SSEA-1) and Octamer-4 (Oct-4) were evaluated before and after cryopreservation. The effects of two methods of cryopreservation, slow freezing and vitrification, were studied. The ES cells were cryopreserved either as single cells or as clumps. The viability of a single cell after slow freezing was 88%, but after vitrification no single cell was recovered. Surviving clumps after slow freezing quickly recovered and exhibited a morphology indistinguishable from noncryopreserved cells. After vitrification, 2 weeks of culture were required for the cells in clumps to proliferate enough for subculturing. Analysis of cloning efficiency and the colonies morphology were based on a mouse colony rating scale and their characteristic. The...
本研究对小鼠胚胎干(ES)细胞R1系进行低温保存,测定细胞活力、形态、菌落数和碱性磷酸酶(AP)活性。此外,我们还对冷冻前后的分期特异性抗原(SSEA-1)和Oct-4 (Oct-4)等转录因子的表达进行了评价。研究了慢速冷冻和玻璃化冷冻两种低温保存方法的效果。胚胎干细胞以单细胞或团块形式冷冻保存。慢速冷冻后单细胞存活率为88%,玻璃化后无单细胞恢复。缓慢冷冻后存活的细胞团块迅速恢复,并表现出与非冷冻保存细胞难以区分的形态。玻璃化后,需要2周的培养,才能使成团的细胞增殖到足以进行传代培养。克隆效率和菌落形态分析采用小鼠菌落分级量表及其特点。…
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引用次数: 7
Progression of Cell and Tissue Vitrification 细胞和组织玻璃化的进展
Pub Date : 2007-04-23 DOI: 10.1089/CPT.2006.9988
J. Baust
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引用次数: 0
A Modified Differential Scanning Calorimetry Method for Determining Water Transport Properties in Biological Cells During the Freezing Process 一种改进的差示扫描量热法测定生物细胞在冷冻过程中的水运输特性
Pub Date : 2007-04-23 DOI: 10.1089/CPT.2006.9991
Xu Han, D. Luo, X. Cui, S. Heimfeld, D. Gao
A modified analytical and experimental method using differential scanning calorimeter (DSC) was applied to determine the cell water transport properties of human erythrocytes during the freezing process. Using DSC, samples containing human erythrocyte cell suspensions of human erythrocytes with different cytocrits were cooled to −40°C at slow cooling rates (5°C/min) after nucleation. It was shown that latent heat release from one unit mass of the cell suspension was a linear function of cytocrits (cell numbers), with a temperature-dependent slope and intercept. Based on the theoretical model, cell volumes were calculated from the slope and intercept at corresponding temperatures. Cell water transport properties (lpg,Ea) were next calculated by curve fitting of the cell volume change during the freezing process. The results revealed that, for human erythrocyte, Lpg (Tr = 273.15 K) is 0.10 ± 0.01 μm/min ċ atm and Ea is 279.1 ± 0.7 kJ/mol at the cooling rate of 5°C/min. For comparison, both DSC and cryomicro...
采用改进的差示扫描量热仪(DSC)分析和实验方法测定了人红细胞在冷冻过程中的细胞水转运特性。用DSC法,将含有不同细胞质的人红细胞悬浮液的样品在成核后以慢速冷却(5°C/min)冷却至- 40°C。结果表明,单位质量细胞悬浮液的潜热释放是细胞数目的线性函数,具有与温度相关的斜率和截距。根据理论模型,从相应温度下的斜率和截距计算细胞体积。然后通过对冷冻过程中细胞体积变化的曲线拟合计算细胞的水输运特性(lpg,Ea)。结果表明,在降温速率为5℃/min时,人红细胞的Lpg (Tr = 273.15 K)为0.10±0.01 μm/min, Ea为279.1±0.7 kJ/mol。为了比较,DSC和低温显微…
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引用次数: 6
The Effect of Temperature Gradients on Stress Development During Cryopreservation via Vitrification. 温度梯度对玻璃化冷冻过程中应力发展的影响。
Pub Date : 2007-01-01 DOI: 10.1089/cpt.2007.9994
Paul S Steif, Matthew C Palastro, Yoed Rabin

This study addresses the problem of thermal stress development in bulky specimens during cryopreservation via vitrification (vitreous means glassy in Latin). While this study is a part of an ongoing effort to associate the developing mechanical stress with the relevant physical properties of the cryopreserved media and to its the thermal history, the current paper focuses exclusively on the role of temperature gradients. Temperature gradients arise due to the high cooling rates necessary to facilitate vitrification; the resulting non-uniform temperature distribution leads to differential thermal strain, possibly resulting in cracking. The cooling rate is assumed constant on the outer surface in this study, and the material properties are assumed constant. It is demonstrated that under these assumptions, mechanical stress develops only when the temperature distribution in the specimen approaches thermal equilibrium at a cryogenic storage temperature. It is shown that the maximum possible stresses for a given cooling rate can be computed with a simple thermo-elastic analysis; these stresses are associated with cooling to sufficiently low temperatures and are independent of the variation of viscosity with temperature. Analytic estimates for these stresses are obtained for several idealized shapes, while finite element analysis is used to determine stresses for geometries used in cryopreservation practice. Stresses that develop under a wider range of storage temperatures are also studied with finite element analysis, and the results are summarized with suitable normalizations. It is found that no stresses arise if cooling ceases above the set-temperature, which defines the transition from viscous-dominated to elastic-dominated behavior; the set-temperature is determined principally by the dependency of viscosity upon temperature. Strategies for rapidly reaching low temperatures and avoiding high stresses are inferred from the results.

本研究解决了通过玻璃化(玻璃体在拉丁语中是玻璃的意思)冷冻保存过程中大块标本的热应力发展问题。虽然这项研究是将发展中的机械应力与低温保存介质的相关物理性质及其热历史联系起来的持续努力的一部分,但目前的论文只关注温度梯度的作用。温度梯度的产生是由于玻璃化所需的高冷却速率;由此产生的不均匀温度分布导致热应变差异,可能导致开裂。本研究假设外表面冷却速率恒定,材料性能恒定。结果表明,在这些假设下,只有当低温储存温度下试样的温度分布接近热平衡时,机械应力才会产生。结果表明,在给定冷却速率下,可以用简单的热弹性分析计算出最大可能应力;这些应力与冷却到足够低的温度有关,与粘度随温度的变化无关。这些应力的分析估计得到了几种理想的形状,而有限元分析是用来确定在低温保存实践中使用的几何应力。通过有限元分析研究了在较宽的储存温度范围内产生的应力,并对结果进行了适当的归一化总结。研究发现,当冷却在设定温度以上停止时,没有应力产生,这定义了从粘为主到弹性为主的转变;设定温度主要取决于粘度对温度的依赖性。从结果中推断出快速达到低温和避免高应力的策略。
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引用次数: 63
International Exchange of Cord Blood: Networks And Logistics 脐带血国际交流:网络和物流
Pub Date : 2006-12-01 DOI: 10.1089/CPT.2006.9995
J. Garcia, M. Torrabadella
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引用次数: 1
期刊
Cell Preservation Technology
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