壳聚糖酶的克隆表达与壳寡糖的制备分析

目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。     

颧骨颧弓骨折的分类及诊治

颧骨颧弓骨折是最常见的面中部骨折之一,迄今为止,在其分类及诊治的选择上仍然存在一定争议,本文对目前颧骨颧弓骨折的分类及诊治的现状和进展作一综述,以期对临床工作提供参考。     

国内外蝗害治理技术现状与展望

本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...     

Dogs and Practices of Community and Neighboring

ABSTRACT

Dogs are important facilitators of social interaction. However, little attention has been given to the specific mechanisms through which these relations proceed, or to the ways that dogs help to broker, maintain, and even disrupt social relations. This paper addresses this absence through an indepth qualitative analysis of the everyday experiences of 24 dog owning households who live in apartments in Sydney, Australia. It shows that dogs encourage people to spend more time outside, make people recognizable within their neighborhood, provide a topic of conversation, and actively solicit the attention of strangers. Dogs help make people recognizable and identifiable to others, while also creating social distance. The paper connects to broader literature on neighboring and community practice to show that community relations shaped by dogs involve practices of inclusion as well as exclusion. Exclusion provides an important motivation for new community formation.     

The isolation and purification of surface specific proteins of somatic and reproductive protoplasts of lily and rapeseed

The plasma membrane surface proteins of intact somatic (leaf) and reproductive (pollen, generative cell or sperm cell) protoplasts of lily ( Lilium longiflorum ) and rapeseed ( Brassica napus cv. Midas) were compared after probing with N-hydroxysuccinimido- (NHS) or sulfo-NHS-biotin. The plasma membranes of intact protoplasts are impermeable to these biotin probes, which bind covalently to the free amino groups of surface proteins. Enzyme-labelled streptavidin was used to detect membrane proteins after separation by SDS-PAGE and western blotting. In lily, six proteins specific to the surface membrane of leaf protoplasts were identified varying from 25–64 kDa, three proteins to pollen protoplasts in the range 35–64 kDa and two proteins to generative cell protoplasts, 63 and 67 kDa. In rapeseed leaf protoplasts, seven proteins in the range 22–69 kDa were detected, while in the sperm enriched fraction five proteins were present in the same kDa range. The proteins identified as membrane specific for generative cell protoplasts of lily have been isolated and were used as antigens for monoclonal antibody production. Preliminary results indicate the successful production of antibodies to surface antigens. These antibodies will be used to localise surface specific epitopes which are likely to be involved in cell-cell recognition at fertilization.     

Microbiological and Biotechnological Aspects of Metabolism of Carbamates and Organophosphates

Abstract

Several carbamate and organophosphate compounds are used to control a wide variety of insect pests, weeds, and disease-transmitting vectors. These chemicals were introduced to replace the recalcitrant and hazardous chlorinated pesticides. Although newly introduced pesticides were considered to be biodegradable, some of them are highly toxic and their residues are found in certain environments. In addition, degradation of some of the carbamates generates metabolites that are also toxic. In general, hydrolysis of the carbamate and organophosphates yields less toxic metabolites compared with the metabolites produced from oxidation. Although microorganisms capable of degrading many of these pesticides have been isolated, knowledge about the biochemical pathways and respective genes involved in the degradation is sparse. Recently, a great deal of interest in the mechanisms of biodegradation of carbamate and organophosphate compounds has been shown because (1) an efficient mineralization of the pesticides used for insect control could eliminate the problems of environmental pollution, (2) a balance between degradation and efficacy of pesticides could result in safer application and effective insect control, and (3) knowledge about the mechanisms of biodegradation could help to deal with situations leading to the generation of toxic metabolites and bioremediation of polluted environments. In addition, advances in genetic engineering and biotechnology offer great potential to exploit the degradative properties of microorganisms in order to develop bioremediation strategies and novel applications such as development of economic plants tolerant to herbicides. In this review, recent advances in the biochemical and genetic aspects of microbial degradation of carbamate and organophosphates are discussed and areas in need of further investigation identified.     

Activity and stability of catalase in blood and tissues of normal and acatalasemic mice

Catalase activity in blood, liver, and kidney of a mutant strain Cs^b has been found to be decreased as compared to the level in normal mice. However, the extent of the reduction largely depends on the conditions used for activity determination, in particular, temperature and duration of the incubation period. In liver, this effect is most pronounced, the observed activity in mutants varying between 21 and 85% of the normal level. This dependence on the assay conditions is mainly due to the unusual heat lability of the variant enzyme, which undergoes rapid inactivation when incubated at 37 C.     

Coiled-coil nanomechanics and uncoiling and unfolding of the superhelix and alpha-helices of myosin

The nanomechanical properties of the coiled-coils of myosin are fundamentally important in understanding muscle assembly and contraction. Force spectra of single molecules of double-headed myosin, single-headed myosin, and coiled-coil tail fragments were acquired with an atomic force microscope and displayed characteristic triphasic force-distance responses to stretch: a rise phase (R) and a plateau phase (P) and an exponential phase (E). The R and P phases arise mainly from the stretching of the coiled-coils, with the hinge region being the main contributor to the rise phase at low force. Only the E phase was analyzable by the worm-like chain model of polymer elasticity. Restrained molecular mechanics simulations on an existing x-ray structure of scallop S2 yielded force spectra with either two or three phases, depending on the mode of stretch. It revealed that coiled-coil chains separate completely near the end of the P phase and the stretching of the unfolded chains gives rise to the E phase. Extensive conformational searching yielded a P phase force near 40 pN that agreed well with the experimental value. We suggest that the flexible and elastic S2 region, particularly the hinge region, may undergo force-induced unfolding and extend reversibly during actomyosin powerstroke.     

硼、镉元素及其交互作用与油菜产量和品质

硼和镉两种元素是影响油菜产量和品质的两个重要因素.硼是植物生长所必需的微量元素,施硼是油菜种植的必需环节;镉是植物生长的非必要元素,易在油菜体内富集,可能通过食物链危害人体健康.本文主要从镉含量与油菜食品安全品质角度考虑,阐述了油菜对镉的积累和耐受机制;同时,概括了前人总结的硼对油菜的产量和品质的影响.最后,结合本人研究区广西地区土壤有效硼含量低,全镉含量高的现状,提出运用硼镉交互作用机理,通过施加适量硼肥,提高油菜的产量和品质,消除土壤镉的潜在危害,从本质上改善该地区土壤存在的低硼高镉现状.     

Phylogeny and taxonomy of Aspicilia and Megasporaceae

Phylogenetic analyses based on nuLSU and mtSSU indicate that Megasporaceae is monophyletic. Aspicilia species were distributed among three main well supported groups and one group with low support that included the type species; a division of the family into five genera is proposed. The old names Circinaria and Sagedia are reintroduced for groups not including A. cinerea, the type of Aspicilia. The monotypic Megaspora is closely related to Circinaria, while Lobothallia is the sister group of the other Megasporaceae genera. Aspicilia recedens and A. farinosa are transferred to Lobothallia. Species of the 'Sphaerothallia group' are nested in Circinaria. Aspilidea is not a member of Megasporaceae but seems to be more closely related to Ochrolechiaceae. Aspilidea myrinii is neotypified, and lectotypes are designated for Aspicilia gibbosa, A. leprosescens and Lecanora gibbosula.     

Effects of husbandry and management systems on physiology and behaviour of farmed and laboratory rabbits

The major issues regarding the welfare of both farmed and laboratory rabbits are reviewed, according to husbandry and management systems. The main stressors that can affect welfare and homeostatic responses in rabbits are also reviewed. An overview of the most widespread housing systems for both farmed and laboratory rabbits is presented. The main problems related to housing and management are identified, in particular those related to individual and group housing, space requirements and group size, as well as human-animal interaction. The effects of psychological and physical stressors on physiology and behaviour are illustrated through examples in various rearing conditions. Psychological stressors include social stress and fear, while physical stressors include environmental variables such as housing system and climatic factors, i.e. heat. Welfare indicators are identified that can be monitored to determine the effects of individual and environmental variables on the animals' possible coping strategies. Physiological indicators include the neuro-endocrine and psycho-neuro-immuno-endocrine measurements, while behavioural indicators include the behavioural repertoire and responses to behavioural tests. Some possible ways to enhance welfare are indicated, such as enrichment of the environment and improved handling procedures.     

Distribution and Functions of Sterols and Sphingolipids

Sterols and sphingolipids are considered mainly eukaryotic lipids even though both are present in some prokaryotes, with sphingolipids being more widespread than sterols. Both sterols and sphingolipids differ in their structural features in vertebrates, plants, and fungi. Interestingly, some invertebrates cannot synthesize sterols de novo and seem to have a reduced dependence on sterols. Sphingolipids and sterols are found in the plasma membrane, but we do not have a clear picture of their precise intracellular localization. Advances in lipidomics and subcellular fractionation should help to improve this situation. Genetic approaches have provided insights into the diversity of sterol and sphingolipid functions in eukaryotes providing evidence that these two lipid classes function together. Intermediates in sphingolipid biosynthesis and degradation are involved in signaling pathways, whereas sterol structures are converted to hormones. Both lipids have been implicated in regulating membrane trafficking.Typical examples of eukaryotic lipids, sterols, and sphingolipids can both be found in membranes from simple unicellular fungi and protists to multicellular animals and plants. Their versatile use as structural elements but also as signaling molecules has probably played an important role during the evolution of this large and diverse group of organisms. There are also many eukaryotes that have lost the ability to synthesize sterols de novo including nematodes, insects, and marine invertebrates, which have to take up sterols with their diet. Sterol biosynthesis has also been reported in a number of bacteria. Sphingolipids are more widely spread among prokaryotes than sterols and also show a greater variety of structures among the different eukaryotes.In this short review we will first give an overview about the diversity of sterol and sphingolipid structures and their distribution in nature. Then we will discuss their subcellular distribution. A brief technical section will add some information on the separation and detection of these lipid molecules. Subsequently, we will summarize different genetic approaches to study the functions of sterols and sphingolipids, and finally, we will discuss the functional and possible physical interactions of the two lipid classes within the cell. Far from being comprehensive, we will focus only on a few interesting aspects and try to give new view points, which are less frequently discussed.     

Biodiversity and the productivity and stability of ecosystems

Attempts to unveil the relationships between the taxonomic diversity, productivity and stability of ecosystems continue to generate inconclusive, contradictory and controversial conclusions. New insights from recent studies support the hypothesis that species diversity enhances productivity and stability in some ecosystems, but not in others. Appreciation is growing for the ways that particular ecosystem features, such as environmental variability and nutrient stress, can influence biotic interactions. Alternatives to the diversity-stability hypothesis have been proposed, and experimental approaches are starting to evolve to test these hypotheses and to elucidate the mechanisms underlying the functional role of species diversity.     

Genetic and physical mapping of telomeres and macrosatellites of rice

Telomeres and telomere-associated satellites of rice were genetically and physically analyzed by pulsed-field gel electrophoresis (PFGE) using Arabidopsis telomeric DNA and rice satellite sequences as probes. We demonstrate that Arabidopsis telomeric sequences hybridize to rice telomeres under the conditions of high stringency. Using the Arabidopsis probe, multiple, discrete telomeric fragments could be identified on pulsed-field gel blots of rice DNAs digested with rare-cutting restriction enzymes. Most of the telomeric bands larger than 300 kb are physically linked with satellite bands as revealed by PFGE. Some of the telomeric and satellite bands segregate in a Mendelian fashion and are highly reproducible. Three such telomeric bands have been mapped to the distal ends of RFLP linkage groups: Telsm-1 on chromosome 8, Telsa-1 on chromosome 9 and Telsm-3 on chromosome 11. One segregating satellite band was mapped to an internal region of chromosome 10. Telomeric fragments were shown not only to be genetically linked to but also physically linked (based on PFGE) to the terminal RFLP markers. The physical distance from telomeric sequences to a distal RFLP marker, r45s gene, on chromosome 9, is 200 kb while the distance from telomeric sequences to RG98, a terminal RFLP marker on chromosome 11, is 260 kb. Physical maps of the telomere regions of chromosome 9 and chromosome 11 are presented.     

Ontogeny and phylogeny of endothermy and torpor in mammals and birds

Endothermic thermoregulation in small, altricial mammals and birds develops at about one third to half of adult size. The small size and consequently high heat loss in these young should result in more pronounced energetic challenges than in adults. Thus, employing torpor (a controlled reduction of metabolic rate and body temperature) during development would allow them to save energy. Although torpor during development in endotherms is likely to occur in many species, it has been documented in only a few. In small, altricial birds (4 orders) and marsupials (1 order), which are poikilothermic at hatching/birth, the development of competent endothermic thermoregulation during cold exposure appears to be concurrent with the capability to display torpor (i.e. poikilothermy is followed by heterothermy), supporting the view that torpor is phylogenetically old and likely plesiomorphic. In contrast, in small, altricial placental mammals (2 orders), poikilothermy at birth is followed first by a homeothermic phase after endothermic thermoregulation is established; the ability to employ torpor develops later (i.e. poikilothermy-homeothermy-heterothermy). This suggests that in placentals torpor is a derived trait that evolved secondarily after a homeothermic phase in certain taxa perhaps as a response to energetic challenges. As mammals and birds arose from different reptilian lineages, endothermy likely evolved separately in the two classes, and given that the developmental sequence of torpor differs between marsupials and placentals, torpor seems to have evolved at least thrice.     

食蟹猴和恒河猴卵母细胞及早期胚胎玻璃化冻存

为了节省经费和使转基因模型动物品种资源得到妥善保存,该研究利用自制的梯度浓度冷冻液和解冻液结合玻璃化方式分别冷冻和解冻了非人灵长类动物的183个卵母细胞（GV期、MI期和MII期）、114个卵裂期胚胎（2-细胞期、4-细胞期和8-细胞期）及25个桑椹期胚胎。其中食蟹猴卵母细胞67个,卵裂期胚胎45个,桑椹期胚胎11个;恒河猴卵母细胞116个,卵裂期胚胎69个,桑椹期胚胎14个。复苏后存活率分别为56/67（83.58%）、36/45（80.00%）、9/11（81.82%）、102/116（87.93%）、55/69（79.71%）和11/14（78.57%）。结果表明,快速玻璃化冷冻法简便且胚胎存活率高,是一种较好的冷冻食蟹猴和恒河猴卵母细胞及胚胎的方法。     

Mechanisms and sites of action of newer angiotensin agonists and antagonists in terms of activity and receptor.

From the myotropic and vasopressor activities of the numerous analogs of angiotensin II, it has been determined that the phenyl group of position 8 possesses the information for biologic response while the aromatic side groups in positions 4 and 6, the guanido group in position 2 and the C-terminal carboxyl are involved in binding to the receptor site. Removal of a side group of the C-terminal phenyalanine yields peptides that bind to the receptor. While many of these have low agonist properties, all have antagonist properties. Modifications in the aromatic side groups affect conformation of the octapeptide. This change may relate to receptor binding but sufficient data are not yet available to determine a correlation pattern. A proposed conformation for angiotensin is given as well as an artist's concept of angiotensin II binding to its membrane receptor utilizing the groups known to be involved in binding. Both angiotensin II and III [des-Asp] angiotensin II stimulate the biosynthesis and release of aldosterone from adrenal glomerulosa cells. Sufficient data are not yet available to determine whether the conversion of angiotensin II to angiotensin III is neccessary for the steroidogenesis activity.