骨固定用聚乳酸/无机纳米粒子复合材料

本论文针对目前用于骨固定和骨修复的聚乳酸／无机纳米粒子复合材料的界面强度低、粒子分散不均匀以及所采用生物活性无机填料粒径较大等缺点，对轻基磷灰石及生物活性玻璃无机纳米粒子的制备、界面改性、粒子的分散、以及复合材料的制备进行了较详细的论述。另外，对材料的力学性能、结晶性能和生物相容性进行了较细统的测试和研究。（1）以磷酸和氢氧化钙为原料在40-80℃的反应条件下制备出了米粒状和棒状的HAP粒子，然后在-50℃的冷冻干燥机中干燥48h，得到白色的HAP粉末。用TEM、SEM、WAXD、FTIR等对所得产物进行了表征。研究结果表明，提高反应温度有利于生成高结晶度的长棒状HAP颗粒。此外，锻烧温度对粒子的形貌和结晶度也有很大的影响，锻烧温度越高，粒子的结晶度就越高，并且，当锻烧温度提高到900℃以上时，HAP粒子的形貌会由长棒形逐渐变成球形。（2）在高纯氢气气氛中，以辛酸亚锡为催化剂的反应条件下使左旋丙交酷开环聚合，直接接枝到HAP的表面，使HAP的粒子表面覆盖一层聚乳酸分子，使HAP的亲油性能得到提高。对表面接枝的轻基磷灰石（g-HAP）用31PMAS-NMR、FTIR、TGA、TEM、SEM和GPC进行了表征。结果表明，用此方法可在HAP表面接枝6％的PLLA。（3）用溶剂法制备了PLLA/g-HAP复合材料，并对其机械性能、结晶性能和生物相容性进行了表征。试验结果表明：与纯HAP相比，g-HAP粒子更容易均匀分散到PLLA基体中，当填料含量达到4％时，PLLAg-HAP复合材料的力学性能达到最好。由Dsc和PoM的实验结果表明，g-HAP粒子在聚合物基体中可以起到异相成核剂的作用。细胞实验结果表明，PLL刀g-HAP复合材料的细胞相容性明显优于纯的PLLA和PLLA/HAP复合材料。（4）以正硅酸乙酷（TEOS）、硝酸钙（Ca(NO3）2）和磷酸氢二按（（NH4）ZHPO4）为原料，利用在酸性溶液中水解，碱性溶液中缩聚沉淀，然后将反应液离心分离，冷冻干燥，最后在马弗炉中锻烧的方法，得到白色的5102-coo-PZos三元生物活性玻璃粉末。SEM和TEM分析结果表明，所得到生物活性玻璃是粒径在40nln左右的球形颗粒，且粒径分布非常均匀。（5）以正硅酸乙酷（TEoS）和硝酸钙（Ca（NO3）2）为原料，利用在酸性溶液中水解，碱性溶液中缩聚沉淀，然后将反应液离心分离，冷冻干燥，最后在马弗炉中锻烧的方法，得到粒径为200nm左右的球形SiO2-CaO二元生物活性玻璃粉末。

沙棘(Hippophae rhamnoides L.)在低温和增强UV-B胁迫下的生态生理反应

植物生长和生产力受到自然界各种形式的生物和非生物胁迫因子的影响。这些胁迫包括低温、高温、盐碱、干旱、洪水、重金属、虫害、病害和紫外线辐射等等。而人类活动大大加剧了这些胁迫所带来的影响。由于人类污染而导致臭氧层衰减以及由此产生的地球表面紫外辐射增强已经成为全球气候变化的一个主要方面。UV-B胁迫，甚至当前的辐射水平，所带来的影响已经引起科学工作者的广泛关注。 为了生存和繁殖，植物不得不面临环境中各种潜在胁迫所带来的负面影响。然而，植物生活型的不可移动性决定了其逃避胁迫的局限性。因此，绝大多数植物都是通过对胁迫作出反应，通过修复或者更新组织来降低伤害。而植物应对环境变化的能力则是由其生长模式的种属特异性和本身的遗传组成所决定。在自然界，植物常常同时面临多种胁迫，这些胁迫所引发的植物反应可能具有叠加、协同或者拮抗作用。沙棘是一种具刺、具有固氮功能的多年生雌雄异株灌木，广泛分布于亚欧大陆的温带地区和亚洲亚热带的高海拔地区。在中国，沙棘常常被用作植被恢复中的先锋树种而大量栽培。本文采用沙棘作为模式植物，试图探索木本植物对低温，UV-B辐射增强以及其与干旱的复合胁迫的响应以及沙棘对这些胁迫响应是否具有种群差异性。 对来自南北两个种群的沙棘进行短日照和低温处理，检测了其在抗寒锻炼和抗寒性发育过程中存在的性别差异。结果表明，短日照和低温都分别能够诱导抗寒锻炼的发生，而两者同时存在对所有实验植株抗寒性的大小具有叠加效应。然而，短日照和低温所诱导的抗寒性在两个种群中都具有性别差异性，雄性植株比雌雄植株对短日照和低温更为敏感。同时，南北种群间也存在差异性，北方种群的植物比南方种群的植物对短日照和低温敏感，从而在短日照下抗寒锻炼的发生时间更早，低温诱导的抗寒性更大。短日照和低温诱导植物增加抗寒性的同时伴随着脱落酸的变化。脱落酸的变化因处理，种群和性别的不同而不同。这些生理反应表明不同的沙棘种群，不同的植株性别对同一环境胁迫可能存在不同的生存策略。 比较了来自高低两个海拔的沙棘种群对于干旱和UV-B辐射增强以及两者复合胁迫条件下的生理生态反应。干旱使两个种群中植株总的生物量，总叶面积，比叶面积，叶片含碳量，含磷量，木质素含量和碳氮比显著降低，使根冠比，粗根细根比和叶片脱落酸含量显著增加。干旱而非UV-B使得δ13C 值显著增加。但是，比较而言，来自高海拔的种群对干旱反应更为强烈，而来自低海拔的种群对UV-B更敏感。在UV-B辐射增强的处理下，干旱所诱导的脱落酸的积累被显著抑制。而且我们检测到在一些指标上存在显著的干旱×UV-B交互作用，如两个种群中在总生物量上，低海拔种群中在总叶面积，粗根细根比上，高海拔种群中在比叶面积，δ13C值，木质素含量上都存在明显的交互作用。这些结果表明这两个种群对胁迫具有不同的适应性反应，来自高海拔的种群比来自低海拔的种群更能够抵御干旱和UV-B胁迫。 室外实验表明，UV-B 去除/增补对沙棘高低两个海拔种群的影响都不大。对生物量的积累，植株高度以及一些常见的胁迫反应生理指标比如丙二醛、ABA 和游离脯氨酸都没有显著影响。UV-B 的效应比UV-A 大，植物反应在无UV 和仅有UV-A 的处理间没有什么区别。然而，UV-B 去除的两个处理和UV-B 存在的两个处理间存在显著区别。UV-B 使得两个种群都显著降低了比叶面积(SLA)，但却使长期用水效率增加。但UV-B对光合色素和光合系统II 的影响不大。总体看来，来自低海拔的种群对UV-B 更为敏感。 Plant is adversely affected by various abiotic and biotic stress factors. These stressors includelow temperature, heat, salt, drought, flooding, heavy metal toxicity, wounding by herbivores,infecting by pathogenic microorganisms, ultraviolet (UV) radiation and so on. Variousanthropogenic activities have accentuated the existing stress factors. One of the mostimportant aspects of global change is that of stratospheric ozone depletion caused by seriousanthropogenic pollution and the resulting increase in UV radiation reaching the surface of theEarth. Scientists have become concerned about the effects that considerable UV-B stress, evenat current levels. In order to survive and reproduce, plants have to be able to cope with lots of potentiallyharmful stress factors that are almost constantly present in their environment. Most plants’responses under stress are to neutralize the stress, repairing the damage or regrowing newtissue rather than to avoid it due to their sessile life style. The plant defense capacity dependson plant-specific modular growth patterns and genetic make-up that allows for flexibleresponses to changing environments. Plants usually encounter several stresses simultaneouslyunder field conditions, and the stresses may cause a variety of plant responses, which can beadditive, synergistic or antagonistic. Sea buckthorn (Hippophae rhamnoides L.), a thorny nitrogen fixing deciduously perennialshrub, which is widely distributed throughout the temperate zones of Asia and Europe and thesubtropical zones of Asia at high altitudes. It has been widely used in forest restoration as thepioneer species in China. In this paper, we used sea buckthorn as a model, tried to get some understand of how plants fight low temperature, enhanced UV-B radiation level and thatcombination of drought. And also, want to know whether does there exist some populationspecific responses to such stressors. Sexual differences in cold acclimation and freezing tolerance development of two contrastingsea buckthorn (Hippophae rhamnoides L.) ecotypes from northern and southern regions inChina were recorded after exposure to short day photoperiod (SD) and low temperature (LT).The results demonstrated that cold acclimation could be triggered by exposing the plants toSD or LT alone, and that a combination of both treatments had an additive effect on freezingtolerance in all plants tested. However, development of freezing tolerance was dependent onthe sex of plants under SD and LT, the males were clearly more responsive to SD and LT thanthe females in both ecotypes studied. On the other hand, development of freezing tolerancewas also ecotype-dependent, the northern ecotype was more responsive to SD and LT than thesouthern ecotype, resulting in earlier cold acclimation under SD and higher freezing toleranceunder LT. Moreover, development of freezing tolerance induced by SD and LT wasaccompanied by changes in ABA levels. These alterations in ABA levels were different indifferent treatments, ecotypes and sexes. Therefore, the differences in SD and LT-inducedphysiological responses showed that the different ecotypes and the different sexes mightemploy different survival strategies under environmental stress. Two contrasting populations from the low and high altitudinal regions were employed toinvestigate the effects of drought, UV-B and their combination on sea buckthorn. Droughtsignificantly decreased total biomass, total leaf area, specific leaf area，leaf carbon (C),phophous (P), lignin content and the ratio of C: N in both populations, and increasedroot/shoot ratio, fine root/coarse root ratio and abscisic acid content (ABA), in bothpopulations. Drought but not UV-B resulted in significantly greater carbon isotopecomposition (δ13C) values in both populations. However, the high altitudinal population wasmore responsive to drought than the low altitudinal population. The drought-inducedenhancement of ABA in the high altitudinal population was significantly suppressed in thecombination of drought and elevated UV-B. Moreover, significant drought × UV-B interactionwas detected on total biomass in both populations, total leaf area and fine root/coarse root inthe low altitudinal population, specific leaf area, δ13C value and leaf lignin content in the high altitudinal population. These results demonstrated that there were different adaptive responsesbetween two contrasting populations, the high altitudinal population exhibited highertolerance to drought and UV-B than the low altitudinal population. A field experiment was conducted to investigate effects of UV-B exclusion/supplementationon two altitudinal populations of sea buckthorn. UV-B exclusion or supplementation had littleeffects on both populations investigated. For instance, the total biomass, plant height andsome physiological index such as Malondialdehyde (MDA), ABA and free proline were notchanged significantly. The UV-B effects are more significant than that of UV-A, nodifferences were found between treatments of excluded UV and excluded UV-B. However,compared with treatments of UV-B exclusion (including absent of UV-B and all UV band),the present of UV-B (including near ambient environment and enhanced UV-B) significantdecreased specific leaf area, and increased long time water use efficiency as evaluated by δ13Cvalue. UV-B had little effects on photosynthetic pigments and Photosystem II (PSII). The lowaltitude population is more sensitive to UV-B than that of the high altitude population.

ACHT操作引起黑麦(Secale cereale)染色体的畸变和重建

用ACHT处理黑麦萌动种子，对修复前后材料的观察和分析结果表明：1. ACHT操作引起染色体数目变化和染色体断裂损失。在一定 条件和范围内，不同处理引起的这种变化具有显著差异，条件越剧烈，染色体数目变化的范围和频率愈大，断片发生的数量和频率 也愈高，同时修复前后染色体数目的变化范围和频率与断片发生的数量和频率以及它们的修复频率均表现明显的相关性。2. ACHT 操作引起染色体畸变的多样性。经ACHT处理后，胚根细胞染色体有4种断裂方式，包括着丝粒断裂、次溢痕断裂、长臂断裂和短臂 断裂等，其中着丝断裂频率最高；产生6种断片类型，包括有着丝粒和端粒的、有着丝粒而无端粒的、有部分着丝粒和端粒的、有 部分着丝粒而无端粒的、只有端粒的、既无着丝粒也无端粒的断片等。3. ACHT操作引起遗传结构重建的多样性。经ACHT处理后， 对修复24-72小时材料进行核型比较(按Stebbins 和 Levan 标准)和随体分析，处理细胞在染色体数目、大小、形态、位置等方面 均发生显著变化，说明ACHT处理使这些细胞的染色体结构和染色体组型发生了深刻变化。进一步通过Giemsa C— 带分析，观察到 多种重建染色体类型，包括易位型染色体、附加型染色体、无着丝粒染色体、化染色体、增加的m染色体以及某些带型特异的染色 体等。4. RAPD 分析从分子水平上验证了ACHT能有效地引起遗传结构的改变。所用10种引物对处理和对照材料基因组DNA的扩增产 物在条带数目、条带位置及带型特征等方面均有明显差异，其中4种引物出现条带减少，6种引物出现条带增加，后者还包括一个带 位移动。这说明两种材料的基因组DNA具有明显的RAPD反应多态性差异。This paper descripes some results draw on the basis of the observation and analysis on the rye before and after repaired through treating its budding seeds by ACHT in contrast to without ACHT: 1. ACHT manipulation caused the number variation and breakage damage of rye chromosome. Within certain conditions and timits, this phenomenon caused by different treats had signifcant difference: the more the treatment condition is drastie, the more the chageable range and frequence of rye chromosomae number, and so is the produced fragments. Meanwhile, there existed striking relationship among the changeable range and frequence of rye chromosome, the produced number and frequence of fragments and repairing frequence. 2. ACHT manipulafion engendered the diversify of rye chromosomal aberration. Four breakage patterns and six sorts of fragment were observed by watching the chromosome of the rye radicle treated by ACHT, including centric breakage (occuring in the highest frequence), secondary constriction breakage, long arm breakage and short arm breakage to the former, Comprising that with both centromere and telomere, that with centromere and without telomere, that with partial centromere and with telomere, that with partfial ceetromere and without telomere, that only with telomere and that neither with centromere nor with telomere, etc. 3. ACHT manipulation engendered the diversify or rye genetic structs reconstruction. Karureotype analysis(according to Stebbins and Levan) and satellite anaeysis were carried out to rye radicle through 24-72-hour-long recoverage after ACHT manipulation, which showed remarkable change happened on the rye chromosomal number、shape、arm ration and pattern, etc. and also on the satellite number、size、shape and location etc. Those indicated that ACHT manipulation engendered violent changes to rye chromatin structure and chromosome type. Further Giemsa C-banding analysis showed many types of reconstructed chromosome, such as translocation、addition、without centromere、st and other chromosome. 4. RAPD analysis checked the validity of ACHT on changing genetic structure of rye on the level of molecular biology. The treated and recovered rye has different amplifying band pattern by using IO valid arbitary primers selected from 40 comparing with the control.

Surface modification of bioactive glass nanoparticles and the mechanical and biological properties of poly(L-lactide) composites

Novel bioactive glass (13G) nanoparticles/poly(L-lactide) (PLLA) composites were prepared as promising bone-repairing materials. The BG nanoparticles (Si:P:Ca = 29:13:58 weight ratio) of about 40 run diameter were prepared via the sol-gel method. In order to improve the phase compatibility between the polymer and the inorganic phase, PLLA (M-n = 9700 Da) was linked to the surface of the BG particles by diisocyanate. The grafting ratio of PLLA was in the vicinity of 20 wt.%. The grafting modification could improve the tensile strength, tensile modulus and impact energy of the composites by increasing the phase compatibility.

In vivo mineralization and osteogenesis of nanocomposite scaffold of poly (lactide-co-glycolide) and hydroxyapatite surface-grafted with poly(L-lactide)

Nanocomposite of hydroxyapatite (HAP) surface-grafted with poly(L-lactide) (PLLA) (g-HAP) shows a wide application for bone fixation materials due to its improved interface compatibility, mechanical property and biocompatibility in our previous study. In this paper, a 3-D porous scaffold of g-HAP/poly (lactide-co-glycolide) (PLGA) was fabricated using the solvent casting/particulate leaching method to investigate its applications in bone replacement and tissue engineering. The composite of un-grafted HAP/PLGA and neat PLGA were used as controls. Their in vivo mineralization and osteogenesis were investigated by intramuscular implantation and replacement for repairing radius defects of rabbits. After surface modification, more uniform distribution of g-HAP particles but a lower calcium exposure on the surface of g-HAP/PLGA was observed. Intramuscular implantation study showed that the scaffold of g-HAP/PLGA was more stable than that of PLGA, and exhibited similar mineralization and biodegradability to HAP/PLGA at the 12-20 weeks post-surgery.

具有随机机器故障的制造过程建模与性能分析

为研究机器故障和维修活动对制造过程性能的影响,提出一种基于广义随机Petri网的制造过程建模与性能分析方法。分析了随机机器故障特征;定义了两种故障发现模式和两种中断作业处理策略;给出具有随机机器故障的制造过程的不同模型方法;通过对模型结构特征的分析,证明了其有效性。针对不同策略和参数设置进行了性能仿真。分别以平均产量和平均过程流时间等性能指标,分析了单个工作站的性能;采用平均产量,分析了具有两个工作站的流水线的性能。仿真结果表明,故障率、平均维修时间、缓存数量配置、维修工人数量、故障发现模式和中断作业处理策略是影响具有随机机器故障的制造过程性能的主要因素。

冀东油田南堡滩海地区油气层损害机理与油气层保护方法研究及应用

Study and Application of Damage Mechanism and Protection Method of reservoir in Nanpu Shallow Beach Sea Area is one of the key research projects of Jidong Oilfield Company of PetroChina Company Limited from 2007 to 2008. Located at Nanpu Sag in Huanghua Depression of Bohaiwan Basin, Nanpu Shallow Beach Sea Area with 1000km2 exploration area posseses three sets, shallow Minghuazhen Formation and Guantao Formation of Upper Tertiary, middle-deep Dongying Formation of Lower Tertiary, deep Ordovician, of oil bearing series, according to the achievement of the connecting 3D seismic structure interpretation and the structural geological comprehensive research. Its main reservoir types include Upper Tertiary structural reservoir, Lower Tertiary structural and lithological-structural reservoir, and Ordovician ancient buried hill reservoir. How to protect reservoir, complete well and lift high efficiently is the key to realize high and stable yield of the oil wells during drilling, completing well, testing and repairing well. It is important for reservoir protecting during drilling that directly relate to efficient exploration. Therefore, beginning with basic characteristics and sensitive analysis of reservoir, study of reservoir damage machinism and analysis of reservoir damage potential factor are emphasized when prediction analysis about three-pressure profiles is carried out. The study both of physical and chemical properties and of the strata of the technology of borehole stabilization and reservoir protecting are outstanding. As the conclusions follow: （1）Based on the laboratory experiment about basalt cores, prediction of three- pressure profiles about 30 wells on No.1 and No.2 structure is practiced. The laws of plane pressure distribution are analyzed. （2）According to the analyses about reservoir feature data and about sensitivity evaluation to damage factor in Nanpu oil field, the scheme of reservoir protecting to the sand reservoir of Guantao Formation and the first section of Dongying Formation is put forward. （3）On basis of the analyses on lithological characteristics, mineral compositions, clay minerals, electrical behavior features, physical and chemical properties of basalt of Guantao formation in No.1 and No.2 structure, instability mechanism of basalt sidewall and technical countermeasures are obtained. （4）Aiming at the characteristics of Ordovician dissolution-pore fracture type carbonate reservoir, the scheme of the reservoir protecting to Ordovician is put forward. Creative study of the film forming and sealing and low invasion reservoir protection drilling fluid are successful. In summary, through the study of reservoir heterogeneity and sensitivity, a set of technology and schemes of reservoir protecting is put forward, which is adaptive during drilling the target bed in the research area and establishes the base for efficient exploration. Significant effect has showed in its application in Nanpu oil field.

Dependency-Directed Localization of Software Bugs

Software bugs are violated specifications. Debugging is the process that culminates in repairing a program so that it satisfies its specification. An important part of debugging is localization, whereby the smallest region of the program that manifests the bug is found. The Debugging Assistant (DEBUSSI) localizes bugs by reasoning about logical dependencies. DEBUSSI manipulates the assumptions that underlie a bug manifestation, eventually localizing the bug to one particular assumption. At the same time, DEBUSSI acquires specification information, thereby extending its understanding of the buggy program. The techniques used for debugging fully implemented code are also appropriate for validating partial designs.

Understanding Simple Picture Programs

What are the characteristics of the process by which an intent is transformed into a plan and then a program? How is a program debugged? This paper analyzes these questions in the context of understanding simple turtle programs. To understand and debug a program, a description of its intent is required. For turtle programs, this is a model of the desired geometric picture. a picture language is provided for this purpose. Annotation is necessary for documenting the performance of a program in such a way that the system can examine the procedures behavior as well as consider hypothetical lines of development due to tentative debugging edits. A descriptive framework representing both causality and teleology is developed. To understand the relation between program and model, the plan must be known. The plan is a description of the methodology for accomplishing the model. Concepts are explicated for translating the global intent of a declarative model into the local imperative code of a program. Given the plan, model and program, the system can interpret the picture and recognize inconsistencies. The description of the discrepancies between the picture actually produced by the program and the intended scene is the input to a debugging system. Repair of the program is based on a combination of general debugging techniques and specific fixing knowledge associated with the geometric model primitives. In both the plan and repairing the bugs, the system exhibits an interesting style of analysis. It is capable of debugging itself and reformulating its analysis of a plan or bug in response to self-criticism. In this fashion, it can qualitatively reformulate its theory of the program or error to account for surprises or anomalies.

Die Organisation und Leitung der Arbeit Mit Audiovisuellen Lehr- und Lernmitteln an der Humboldt-Universität zu Berlin und an Anderen Hochschulen in der Ddr

Heun Hans-Georg, Die Organisation und Ieitung der Arbeit mit audiovisuellen lehr- und Ißrnmitteln an der Humboldt-Universität zu Berlin und an anderen Hochschulen in der DDR (Organization and conducting of work with audiovidual aids at the Humboldt University and other schools of higher education in the German Democratic Republic), „Neodidagmata” XX, Poznan 1991, Adam Mickiewicz University Press, pp. 131-136. ISBN 83-232-0302-4. ISBN 0077-653X. Received: November 1987. Audiovisual aids have become now an indispensable help for teachers. The centre of audiovisual aids at the Humboldt University in Berlin performs the following tasks: a) production of audiovisual materials; b) technical service of classes and lectures and seminars and repairing of the equipment; c) information on new audiovisual aids and lending services; d) training of students and university teachers in operating audiovisual aids; e) studies on application of audiovisual aids in schools of higher education.

Zinc finger nuclease gene repair as a treatment for cystinosis

Cystinosis is a multi-system autosomal recessive disorder caused by mutations and/or deletions in both alleles of CTNS, a gene encoding for the low pH dependent lysosomal cystine exporter cystinosin. Cystinosis occurs in approximately 1:200,000 newborns worldwide and is characterised by an accumulation of cystine in the lysosomes. The most severe form of the disorder is nephropathic cystinosis presenting Fanconi syndrome and leads without treatment to an end-stage renal failure before the age of ten. The only treatment available so far is cysteamine therapy, which delays disease progression by five years, but does not provide a cure for cystinosis patients. Current gene and cell based therapeutic approaches have not yet provided a suitable alternative. A potentially approach for a long-term treatment could be to generate autologous gene–modified stem cells by repairing the gene. Zinc Finger Nucleases (ZFNs) serve as a tool to increase HDR up to a 200,000-fold by introducing a double-stranded break (DSB). Thus, simple mutations in the CTNS gene could be corrected by introduction of a double-stranded break using ZFNs to boost the process of HDR with a suitable donor DNA sequence. A permanent repair of the most common lesion CTNS, a 57 kb deletion, could be achieved by ZFN-mediated HDR using a minigene CTNS promoter/cDNA construct. The thesis describes the design and testing of seven zinc finger nuclease pairs for their cleavage activity in vitro and in cellulo.. A highly sensitive assay to detect even low levels of ZFN-mediated HDR was also developed. Finally, to further investigate the role of autophagy in tissue injury in cystinotic cells an assay to monitor autophagy levels in the cells was successfully developed. This assay provides the opportunity to demonstrate functional restoration of CTNS after successful ZFN-HDR in cystinotic cells.

Jackson v JH Watson Property Investment Ltd.

Comments on the Chancery Division decision in Jackson v JH Watson Property Investment Ltd on whether a landlord was liable in nuisance to a long leaseholder in respect of damage caused to the demised property by a building defect which pre-dated the grant of the lease or whether the principle of caveat lessee applied. Considers whether the defect amounted to "disrepair" within the meaning of the landlord's repairing covenant.

Histone H2AX phosphorylation as a molecular pharmacological marker for DNA interstrand crosslink cancer chemotherapy

The aims of this study were to investigate mechanisms of action involved in H2AX phosphorylation by DNA interstrand crosslinking (ICL) agents and determine whether gamma H2AX could be a suitable pharmacological marker for identifying potential ICL cellular chemosensitivity. In normal human fibroblasts, after treatment with nitrogen mustard (HN2) or cisplatin, the peak gamma H2AX response was detected 2-3 h after the peak of DNA ICLs measured using the comet assay, a validated method for detecting ICLs in vitro or in clinical samples. Detection of gamma H2AX foci by immunofluorescence microscopy could be routinely detected with 6-10 times lower concentrations of both drugs compared to detection of ICLs using the comet assay. A major pathway for repairing DNA ICLs is the initial unhooking of the ICL by the ERCC1-XPF endonuclease followed by homologous recombination. HN2 or cisplatin-induced gamma H2AX foci persisted significantly longer in both, ERCC1 or XRCC3 (homologous recombination) defective Chinese hamster cells that are highly sensitive to cell killing by ICL agents compared to wild type or ionising radiation sensitive XRCC5 cells. An advantage of using gamma H2AX immunofluorescence over the comet assay is that it appears to detect ICL chemosensitivity in both ERCC1 and HR defective cells. With HN2 and cisplatin, gamma H2AX foci also persisted in chemosensitive human ovarian cancer cells (A2780) compared to chemoresistant (A2780cisR) cells. These results show that gamma H2AX can act as a highly sensitive and general marker of DNA damage induced by HN2 or cisplatin and shows promise for predicting potential cellular chemosensitivity to ICL agents. (c) 2008 Elsevier Inc. All rights reserved.

Derivation of clinically compliant MSCs from CD105+, CD24-differentiated human ESCs

Adult tissue-derived mesenchymal stem cells ( MSCs) have demonstrated therapeutic efficacy in treating diseases or repairing damaged tissues through mechanisms thought to be mediated by either cell replacement or secretion of paracrine factors. Characterized, self- renewing human ESCs could potentially be an invariable source of consistently uniform MSCs for therapeutic applications. Here we describe a clinically relevant and reproducible manner of generating identical batches of hESC- derived MSC ( hESC- MSC) cultures that circumvents exposure to virus, mouse cells, or serum. Trypsinization and propagation of HuES9 or H1 hESCs in feeder- and serum-free selection media generated three polyclonal, karyotypically stable, and phenotypically MSC-like cultures that do not express pluripotency- associated markers but displayed MSC- like surface antigens and gene expression profile. They differentiate into adipocytes, osteocytes, and chondrocytes in vitro. Gene expression and fluorescence- activated cell sorter analysis identified CD105 and CD24 as highly expressed antigens on hESC- MSCs and hESCs, respectively. CD105+, CD24- monoclonal isolates have a typical MSC gene expression profiles and were identical to each other with a highly correlated gene expression profile ( r(2) >.90). We have developed a protocol to reproducibly generate clinically compliant and identical hESC- MSC cultures.

Microcomputed tomography imaging in a rat model of delayed union/non-union fracture

We aimed to develop a clinically relevant delayed union/non-union fracture model to evaluate a cell therapy intervention repair strategy. Histology, three-dimensional (3D) micro-computed tomography (micro-CT) imaging and mechanical testing were utilized to develop an analytical protocol for qualitative and quantitative assessment of fracture repair. An open femoral diaphyseal osteotomy, combined with periosteal diathermy and endosteal excision, was held in compression by a four pin unilateral external fixator. Three delayed union/non-union fracture groups established at 6 weeks-(a) a control group, (b) a cell therapy group, and (c) a group receiving phosphate-buffered saline (PBS) injection alone-were examined subsequently at 8 and 14 weeks. The histological response was combined fibrous and cartilaginous non-unions in groups A and B with fibrous non-unions in group C. Mineralized callus volume/total volume percentage showed no statistically significant differences between groups. Endosteal calcified tissue volume/endosteal tissue volume, at the center of the fracture site, displayed statistically significant differences between 8 and 14 weeks for cell and PBS intervention groups but not for the control group. The percentage load to failure was significantly lower in the control and cell treatment groups than in the PBS alone group. High-resolution micro-CT imaging provides a powerful tool to augment characterization of repair in delayed union/non-union fractures together with outcomes such as histology and mechanical strength measurement. Accurate, nondestructive, 3D identification of mineralization progression in repairing fractures is enabled in the presence or absence of intervention strategies. (c) 2007 Orthopaedic Research Society.