I.小麦非叶器官碳酸酐酶活性与定位分析II.镉毒害导致芦苇胚性细胞死亡的激素调控研究

第一部分： 通过生理测定和化学染色分析了冬小麦品种小堰54和京411的叶片和非叶片组织的碳酸酐 酶活性。叶片碳酸酐酶活性(CA)在挑旗时期达到最大值，之后减少到最小，而在饱粒期又呈 增加趋势。从灌浆期到饱粒期，颖片和内稃的CA活性均减少，而外稃和种皮的CA活性均增加。在饱粒期，小堰54的叶片、颖片、外稃和种皮CA活性均高于京411。组织化学染色表明，CA主要分布在旗叶的叶肉细胞叶绿体中，也分布在非叶片组织颖片、外稃和内稃的叶肉和维管束鞘细胞的胞质中。这些结果表明，小麦非叶组织叶肉和维管束鞘细胞的胞质中的CA可能对饱粒期冬小麦的C4光合途径起作用。饱粒期小堰54的C02传递到Rubisco酶速率和抗旱性较京411高。 第二部分： 以继代培养的芦苇胚性细胞为材料，利用台盼兰拒染法检测了悬浮细胞死亡过程，并利用石蜡切片法及苏木精染色法观察了不同浓度镉对芦苇细胞的毒害作用。1000μM的CdCl2迅速导致芦苇悬浮细胞死亡，200μM的CdClz在接种后第5天引起悬浮细胞死亡，100μM的CdCl2在接种后第7天引起悬浮细胞死亡，≤50μM的CdClz在接种后7天不引起悬浮细胞死亡。同时对不同浓度镉处理的芦苇胚性细胞的内源植物激素和可溶性蛋白质进行分析，≤50μM的镉浓度显著地降低胚性细胞内IAA、ZR、GA3和GA4的含量，却提高ABA的含量，抑制可溶性蛋白质的合成：≥100μM的镉浓度显著地提高IAA、ZR、GA3和GA4的含量，却降低ABA的含量，促进可溶性蛋白质的合成。这些结果表明，镉的毒害至少包括镉浓度决定的两种细胞死亡机制，高浓度的镉(1000μM)引起的细胞死亡应当为坏死，而100μM的镉引起植物悬浮细胞发生程序性死亡。在较高浓度(≥100μM)的镉处理下，芦苇细胞内内源IAA、ZR、GA3和GA4的浓度较高，可能调控可溶性蛋白质的合成而促进细胞发生程序化死亡。

Cloning, characterization, and molecular application of a beta-agarase gene from Vibrio sp strain V134

V134, a marine isolate of the Vibrio genus, was found to produce a new beta-agarase of the GH16 family. The relevant agarase gene agaV was cloned from V134 and conditionally expressed in Escherichia coli. Enzyme activity analysis revealed that the optimum temperature and pH for the purified recombinant agarase were around 40 degrees C and 7.0. AgaV was demonstrated to be useful in two aspects: first, as an agarolytic enzyme, the purified recombinant AgaV could be employed in the recovery of DNA from agarose gels; second, as a secretion protein, AgaV was explored at the genetic level and used as a reporter in the construction of a secretion signal trap which proved to be a simple and efficient molecular tool for the selection of genes encoding secretion proteins from both gram-positive and gram-negative bacteria.

Cloning, characterization, and molecular application of a beta-agarase gene from Vibrio sp strain V134

V134, a marine isolate of the Vibrio genus, was found to produce a new beta-agarase of the GH16 family. The relevant agarase gene agaV was cloned from V134 and conditionally expressed in Escherichia coli. Enzyme activity analysis revealed that the optimum temperature and pH for the purified recombinant agarase were around 40 degrees C and 7.0. AgaV was demonstrated to be useful in two aspects: first, as an agarolytic enzyme, the purified recombinant AgaV could be employed in the recovery of DNA from agarose gels; second, as a secretion protein, AgaV was explored at the genetic level and used as a reporter in the construction of a secretion signal trap which proved to be a simple and efficient molecular tool for the selection of genes encoding secretion proteins from both gram-positive and gram-negative bacteria.

Intergeneric conjugation in holomycin-producing marine Streptomyces sp strain M095

Marine Streptomyces are potential candidates for novel natural products and industrial catalysts. In order to set up biosynthesis approach for a holomycin-producing strain M095 isotated from Jiaozhou Bay, China, a genetic transformation system was established using intergeneric conjugation. The plasmid pIJ8600 consists of an origin of replication for Escherichia coli, a phage integrase directing efficient site-specific integration in bacterial chromosome, thiostrepton-induced promoter and an attP sequence. Using E. coli ET12567 (pUZ8002) carrying pIJ8600 as a conjugal donor, while it was mated with strain M095, pIJ8600 was mobilized to the recipient and the transferred DNA was also integrated into the recipient chromosome. The frequency of exconjugants was 1.9 +/- 0.13 x 10(-4) per recipient cell. Analysis of eight exconjugants showed pIJ8600 was stable integrated at a single chromosomal site (attB) of the Streptomyces genome. The DNA sequence of the attB was cloned and shown to be conserved. The results of growth and antimicrobial activity analysis indicated that the integration of pIJ8600 did not seem to affect the biosynthesis of antibiotics or other essential amino acids. To demonstrate the feasibility of above gene transfer system, the allophycocyanin gene (apc) from cyanobacterium Anacystis nidulans UTEX625 was expressed in strain M095, and the results indicated heterologous allophycocyanin could be expressed and folded effectively. (c) 2006 Elsevier GmbH. All rights reserved.

attB site disruption in marine Actinomyces sp M048 via DNA transformation of a site-specific integration vector

An efficient conjugation method has been developed for the marine Actinomyces sp. isolate M048 to facilitate the genetic manipulation of the chandrananimycin biosynthesis gene cluster. A phi C31-derived integration vector pIJ8600 containing oriT and attP fragments was introduced into strain M048 by bi-parental conjugation from Escherichia coli ET12567 to strain M048. Transformation efficiency was (6.38 +/- 0.41) x 10(-5) exconjugants per recipient spore. Analysis of eight exconjugants showed that the plasmid pIJ8600 was stably integrated at a single chromosomal site (attB) of the Actinomyces genome. The DNA sequence of the attB was cloned and shown to be conserved. The results of antimicrobial activity analysis indicated that the insertion of plasmid pIJ8600 seemed to affect the biosynthesis of antibiotics that could strongly inhibit the growth of E. coli and Mucor miehei (Tu284). HPLC-MS analysis of the extracts indicated that disruption of the attB site resulted in the complete abolition of chandrananimycin A-C production, proving the identity of the gene cluster. Instead of chandrananimycins, two bafilomycins were produced through disruption of the attB site from the chromosomal DNA of marine Actinomyces sp. M048.

海洋红藻多管藻和松节藻的化学成分及抗氧化活性研究

海洋生物具有产生丰富多样的次生代谢产物的能力，其中红藻门松节藻科海藻卤代次生代谢产物以其结构新颖、生物活性独特引起了天然产物化学家的重视。 本论文对海洋红藻多管藻和松节藻进行了化学成分研究，综合利用各种色谱学方法 (硅胶柱层析、反相硅胶柱层析、凝胶Sephadex LH-20柱层析、半制备高效液相色谱以及重结晶等) 和现代波谱学技术 (IR、UV、EI-MS、FAB-MS、HR-ESI-MS、CD、1H-NMR、13C-NMR、DEPT、1H-1H COSY、HSQC、HMBC等)，共分离鉴定了100个化合物，发现25个新化合物。 从多管藻中分离鉴定38个化合物 (24个溴酚化合物)，其中7个新化合物 (均为溴酚化合物)，包括1个菲并呋喃结构溴酚 (P1), 2个二氢菲结构溴酚 (P2, P3)，1个含 5,7-dihydrodibenzo[c,e]oxepine 结构溴酚 (P4)和3个简单溴酚 (P5, P6, P7)。P1 (urceolatin) 属首例报道的具有菲并呋喃结构的天然产物，从该种中分离的化合物P12 和 P13 可能是其生源合成的前体。P2和P3为第二例报道的具有二氢菲结构的溴酚化合物。 从松节藻中分离并鉴定了62 个化合物，其中18 个为溴酚类新化合物，44 个为已知化合物。化合物具有多变的取代基团，包括2 个脲基吡咯烷酮溴酚化合物 (R1, R2), 4 个γ-脲基丁酸溴酚化合物 (R3-R6)，5 个酰胺溴酚化合物 (R7, R8, R9, R13, R14)，1 个溴酚砜化合物 (R12), 1 个Xanthene 溴酚化合物 (R10)和5 个简单溴酚化合物 (R11, R15, R16, R17, R18)。R1、R2 是首例报道的含有脲基吡咯烷酮片段的天然产物，R10 为首次报道的溴代Xanthene 类天然产物。 对分离到的化合物进行了清除DPPH 和ABTS两种自由基活性的筛选。结果发现溴酚类天然产物具有显著的DPPH自由基清除活性，其中R3 的IC50 仅为3.3 μM, 其活性强度约为阳性对照BHT (IC50 为82.1 μM) 的24倍。另外，溴酚类天然产物对ABTS自由基有较强的清除活性，R2 的TEAC（Trolox efficency activity capacity）值为5.2 mM，约为阳性对照 (ascorbic acid, 1.02 mM) 的 5 倍。初步的构效关系研究发现，稠环分子、多羟基和邻位甲氧基等结构特点能有效增强DPPH 自由基清除活性；特殊取代基如脲基、吡咯烷酮等含有氮原子的基团，能有效增强ABTS 自由基清除活性，多羟基、溴代等结构特点也使其活性有所增强。 本研究结果丰富了海藻卤代化合物的结构类型，为多管藻和松节藻的合理利用提供了一定的科学依据。

Inspection Methods in Programming

The work reported here lies in the area of overlap between artificial intelligence software engineering. As research in artificial intelligence, it is a step towards a model of problem solving in the domain of programming. In particular, this work focuses on the routine aspects of programming which involve the application of previous experience with similar programs. I call this programming by inspection. Programming is viewed here as a kind of engineering activity. Analysis and synthesis by inspection area prominent part of expert problem solving in many other engineering disciplines, such as electrical and mechanical engineering. The notion of inspections methods in programming developed in this work is motivated by similar notions in other areas of engineering. This work is also motivated by current practical concerns in the area of software engineering. The inadequacy of current programming technology is universally recognized. Part of the solution to this problem will be to increase the level of automation in programming. I believe that the next major step in the evolution of more automated programming will be interactive systems which provide a mixture of partially automated program analysis, synthesis and verification. One such system being developed at MIT, called the programmer's apprentice, is the immediate intended application of this work. This report concentrates on the knowledge are of the programmer's apprentice, which is the form of a taxonomy of commonly used algorithms and data structures. To the extent that a programmer is able to construct and manipulate programs in terms of the forms in such a taxonomy, he may relieve himself of many details and generally raise the conceptual level of his interaction with the system, as compared with present day programming environments. Also, since it is practical to expand a great deal of effort pre-analyzing the entries in a library, the difficulty of verifying the correctness of programs constructed this way is correspondingly reduced. The feasibility of this approach is demonstrated by the design of an initial library of common techniques for manipulating symbolic data. This document also reports on the further development of a formalism called the plan calculus for specifying computations in a programming language independent manner. This formalism combines both data and control abstraction in a uniform framework that has facilities for representing multiple points of view and side effects.

Paleoreconstruction of estuarine sediments reveal human-induced weakening of coastal carbon sinks

Human activities in coastal areas frequently cause loss of benthic macrophytes (e.g. seagrasses) and concomitant increases in microalgal production through eutrophication. Whether such changes translate into shifts in the composition of sediment detritus is largely unknown, yet such changes could impact the role these ecosystems play in sequestrating CO 2. We reconstructed the sedimentary records of cores taken from two sites within Botany Bay, Sydney - the site of European settlement of Australia - to look for human-induced changes in dominant sources of detritus in this estuary. Cores covered a period from the present day back to the middle Holocene (~6000 years) according to 210Pb profiles and radiocarbon ( 14C) dating. Depositional histories at both sites could not be characterized by a linear sedimentation rate; sedimentation rates in the last 30-50 years were considerably higher than during the rest of the Holocene. C : N ratios declined and began to exhibit a microalgal source signature from around the time of European settlement, which could be explained by increased nutrient flows into the Bay caused by anthropogenic activity. Analysis of stable isotopic ratios of 12C/ 13C showed that the relative contribution of seagrass and C 3 terrestrial plants (mangroves, saltmarsh) to detritus declined around the time of rapid industrial expansion (~1950s), coinciding with an increase in the contribution of microalgal sources. We conclude that the relative contribution of microalgae to detritus has increased within Botany Bay, and that this shift is the sign of increased industrialization and concomitant eutrophication. Given the lower carbon burial efficiencies of microalgae (~0.1%) relative to seagrasses and C 3 terrestrial plants (up to 10%), such changes represent a substantial weakening of the carbon sink potential of Botany Bay - this occurrence is likely to be common to human-impacted estuaries, and has consequences for the role these systems play in helping to mitigate climate change. © 2011 Blackwell Publishing Ltd.

Classification of team sport activities using a single wearable tracking device

Wearable tracking devices incorporating accelerometers and gyroscopes are increasingly being used for activity analysis in sports. However, minimal research exists relating to their ability to classify common activities. The purpose of this study was to determine whether data obtained from a single wearable tracking device can be used to classify team sport-related activities. Seventy-six non-elite sporting participants were tested during a simulated team sport circuit (involving stationary, walking, jogging, running, changing direction, counter-movement jumping, jumping for distance and tackling activities) in a laboratory setting. A MinimaxX S4 wearable tracking device was worn below the neck, in-line and dorsal to the first to fifth thoracic vertebrae of the spine, with tri-axial accelerometer and gyroscope data collected at 100Hz. Multiple time domain, frequency domain and custom features were extracted from each sensor using 0.5, 1.0, and 1.5s movement capture durations. Features were further screened using a combination of ANOVA and Lasso methods. Relevant features were used to classify the eight activities performed using the Random Forest (RF), Support Vector Machine (SVM) and Logistic Model Tree (LMT) algorithms. The LMT (79-92% classification accuracy) outperformed RF (32-43%) and SVM algorithms (27-40%), obtaining strongest performance using the full model (accelerometer and gyroscope inputs). Processing time can be reduced through feature selection methods (range 1.5-30.2%), however a trade-off exists between classification accuracy and processing time. Movement capture duration also had little impact on classification accuracy or processing time. In sporting scenarios where wearable tracking devices are employed, it is both possible and feasible to accurately classify team sport-related activities.

Oportunidades de melhoria ergonômica das exigências físicas da atividade jangadeira em Ponta Negra Natal-RN

The fishing is an activity with rafts marked by unpredictability, poor tools, physical stress, awkward postures, risk of accidents and incidents, which contribute to insecurity in the context of activity. This study aimed to contribute to reducing of physical effort and the biomechanical impacts in jangadeiro activity of artisanal fishing using jangadas on the beach of Ponta Negra, Natal-RN. To achieve this purpose the methodology of the Work Ergonomic Analysis - WEA became a reference using techniques such as observational and interactional observation notes, photographs, videos, action conversational and listening verbalizations. The resulting activity in health jangadeiro, discussed in this research were: static postural assessment, flexibility of movement of shoulder flexion-extension, flexibility of the hamstrings and lower back, grip and musculoskeletal pain. It was found from observations, interactions and activity analysis, that in carrying out fishing rafts are frequent awkward postures associated with demand for power and many are already feeling the jangadeiros reflections of years of life dedicated to this work. These have already adopted some measures to manage the activity, such as job rotation during the expedition of capture. Therefore, there is a need to expand these measures of activity management, as well as design and capabilities of the raft, seeking positive changes in the activity and consequently the quality of life jangadeiro

Work and technological innovation in organic agriculture

Organic agriculture is a sustainable cultivation ecologically, economically and socially. Several researches in organic agriculture have been made from technical perspectives, economic traits or related to ecological aspects. There are practically no investigations into the nature of the technology used in organic agriculture, especially from an ergonomic perspective. From the activity analysis, this study aimed to map the technology used in the production of organic vegetables. Properties producing organic vegetables were selected representing the State of São Paulo. It was applied an instrument (questionnaire and semi-structured interview) with their managers and it was made visual records to identify adaptations, innovations and technological demands that simultaneously minimize the workload and the difficulties in performing the tasks and increase work productivity. For some of the technological innovations a digital scanner was used to generate a virtual solid model to facilitate its redesign and virtual prototyping. The main results show that organic farmers have little technology in product form. The main innovations that enable competitive advantage or allow higher labor productivity occur in the form of processes, organization and marketing.

Atividade antibacteriana e citotoxicidade do agregado de trióxido mineral associado a diferentes concentrações de clorexidina

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Caracterização do surimi de tilápia do Nilo: morfologia e propiedades físicas, químicas e sensoriais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Produção de óleo essencial associado à deficiência hídrica em plantas Ocimum basilicum L. cv. genovese

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Metabolismo da videira ‘syrah’ no semiárido nordestino sob três estratégias hídricas

The almost complete absence or misdistribution of water as a natural resource frequently constitute a limiting factor for plant growth and development in the semi-arid northeastern Brazil. In this context, the use of appropriate irrigation techniques is an essential and indispensable factor for proper functioning of the primary and secondary metabolisms in plants. This study aimed to assess the metabolism of the Syrah grapevine in the semi-arid northeastern Brazil, by using three irrigation strategies (controlled deficit irrigation [CDI], deficit irrigation [DI], and full irrigation [FI]). The research was conducted at Embrapa Semiarid (Embrapa-Brazilian Agricultural Research Corporation), Experimental Field of Bebedouro, municipality of Petrolina, state of Pernambuco, Brazil. The statistical design was randomized blocks, with a 3 x 6 factorial scheme. Six samplings were performed throughout the experiment cycle at 54, 60, 73, 87, 101, and 115 days after pruning (DAP). We analyzed the total soluble sugars, reducing sugars, total soluble protein, and invertase activity. Analysis of variance and the F test were performed for all analyzed variables. The means were compared using the Tukey test at 5% significance. At the end of the experiment cycle, DI was found to increase the average acid invertase activity in the plant cell wall. Further, by the end of the experiment cycle, the total soluble sugars and reducing sugars increased in all plants in the three irrigation strategies. Thus, we conclude that it is possible to reduce water consumption in vineyards of the semi-arid northeastern Brazil, without significantly affecting the protein and sugar metabolisms in the plants.