A spectral element for wave propagation in honeycomb sandwich construction considering core flexibility

Spectral elements are found to be extremely resourceful to study the wave propagation characteristics of structures at high frequencies. Most of the aerospace structures use honeycomb sandwich constructions. The existing spectral elements use single layer theories for a sandwich construction wherein the two face sheets vibrate together and this model is sufficient for low frequency excitations. At high frequencies, the two face sheets vibrate independently. The Extended Higher order SAndwich Plate theory (EHSaPT) is suitable for representing the independent motion of the face sheets. A 1D spectral element based on EHSaPT is developed in this work. The wave number and the wave speed characteristics are obtained using the developed spectral element. It is shown that the developed spectral element is capable of representing independent wave motions of the face sheets. The propagation speeds of a high frequency modulated pulse in the face sheets and the core of a honeycomb sandwich are demonstrated. Responses of a typical honeycomb sandwich beam to high frequency shock loads are obtained using the developed spectral element and the response match very well with the finite element results. It is shown that the developed spectral element is able to represent the flexibility of the core resulting into independent wave motions in the face sheets, for which a finite element method needs huge degrees of freedom. (C) 2015 Elsevier Ltd. All rights reserved.

Model driven product line engineering : core asset and process implications

Reuse is at the heart of major improvements in productivity and quality in Software Engineering. Both Model Driven Engineering (MDE) and Software Product Line Engineering (SPLE) are software development paradigms that promote reuse. Specifically, they promote systematic reuse and a departure from craftsmanship towards an industrialization of the software development process. MDE and SPLE have established their benefits separately. Their combination, here called Model Driven Product Line Engineering (MDPLE), gathers together the advantages of both. Nevertheless, this blending requires MDE to be recasted in SPLE terms. This has implications on both the core assets and the software development process. The challenges are twofold: (i) models become central core assets from which products are obtained and (ii) the software development process needs to cater for the changes that SPLE and MDE introduce. This dissertation proposes a solution to the first challenge following a feature oriented approach, with an emphasis on reuse and early detection of inconsistencies. The second part is dedicated to assembly processes, a clear example of the complexity MDPLE introduces in software development processes. This work advocates for a new discipline inside the general software development process, i.e., the Assembly Plan Management, which raises the abstraction level and increases reuse in such processes. Different case studies illustrate the presented ideas.

Support for Integrated Ecosystem Assessments of NOAA’s National Estuarine Research Reserves System (NERRS), Volume I: The Impacts of Coastal Development on the Ecology and Human Well-being of Tidal Creek Ecosystems of the US Southeast

A study was conducted, in association with the Sapelo Island and North Carolina National Estuarine Research Reserves (NERRs), to evaluate the impacts of coastal development on sentinel habitats (e.g., tidal creek ecosystems), including potential impacts to human health and well-being. Uplands associated with southeastern tidal creeks and the salt marshes they drain are popular locations for building homes, resorts, and recreational facilities because of the high quality of life and mild climate associated with these environments. Tidal creeks form part of the estuarine ecosystem characterized by high biological productivity, great ecological value, complex environmental gradients, and numerous interconnected processes. This research combined a watershed-level study integrating ecological, public health and human dimension attributes with watershed-level land use data. The approach used for this research was based upon a comparative watershed and ecosystem approach that sampled tidal creek networks draining developed watersheds (e.g., suburban, urban, and industrial) as well as undeveloped sites. The primary objective of this work was to clearly define the relationships between coastal development with its concomitant land use changes and non-point source pollution loading and the ecological and human health and well-being status of tidal creek ecosystems. Nineteen tidal creek systems, located along the southeastern United States coast from southern North Carolina to southern Georgia, were sampled during summer (June-August), 2005 and 2006. Within each system, creeks were divided into two primary segments based upon tidal zoning: intertidal (i.e., shallow, narrow headwater sections) and subtidal (i.e., deeper and wider sections), and watersheds were delineated for each segment. In total, we report findings on 24 intertidal and 19 subtidal creeks. Indicators sampled throughout each creek included water quality (e.g., dissolved oxygen concentration, salinity, nutrients, chlorophyll-a levels), sediment quality (e.g., characteristics, contaminants levels including emerging contaminants), pathogen and viral indicators, and abundance and genetic responses of biological resources (e.g., macrobenthic and nektonic communities, shellfish tissue contaminants, oyster microarray responses). For many indicators, the intertidally-dominated or headwater portions of tidal creeks were found to respond differently than the subtidally-dominated or larger and deeper portions of tidal creeks. Study results indicate that the integrity and productivity of headwater tidal creeks were impaired by land use changes and associated non-point source pollution, suggesting these habitats are valuable early warning sentinels of ensuing ecological impacts and potential public health threats. For these headwater creeks, this research has assisted the validation of a previously developed conceptual model for the southeastern US region. This conceptual model identified adverse changes that generally occurred in the physical and chemical environment (e.g., water quality indicators such as indicator bacteria for sewage pollution or sediment chemical contamination) when impervious cover levels in the watershed reach 10-20%. Ecological characteristics responded and were generally impaired when impervious cover levels exceed 20-30%. Estimates of impervious cover levels defining where human uses are impaired are currently being determined, but it appears that shellfish bed closures and the flooding vulnerability of headwater regions become a concern when impervious cover values exceed 10-30%. This information can be used to forecast the impacts of changing land use patterns on tidal creek environmental quality as well as associated human health and well-being. In addition, this study applied tools and technologies that are adaptable, transferable, and repeatable among the high quality NERRS sites as comparable reference entities to other nearby developed coastal watersheds. The findings herein will be of value in addressing local, regional and national needs for understanding multiple stressor (anthropogenic and human impacts) effects upon estuarine ecosystems and response trends in ecosystem condition with changing coastal impacts (i.e., development, climate change). (PDF contaions 88 pages)

Technologies and Methodologies for the Detection of Harmful Algae and Their Toxins, St Petersburg, Florida, Oct 22-24 2008 : workshop proceedings

The Alliance for Coastal Technologies (ACT) Workshop "Technologies and Methodologies for the Detection of Harmful Algae and their Toxins" convened in St. Petersburg, Florida, October 22- 24, 2008 and was co-sponsored by ACT (http://act-us.info); the Cooperative Institute for Coastal and Estuarine Environmental Technology (CICEET, http://ciceet.unh.edu); and the Florida Fish and Wildlife Conservation Commission (FWC, http://www.myfwc.com). Participants from various sectors, including researchers, coastal decision makers, and technology vendors, collaborated to exchange information and build consensus. They focused on the status of currently available detection technologies and methodologies for harmful algae (HA) and their toxins, provided direction for developing operational use of existing technology, and addressed requirements for future technology developments in this area. Harmful algal blooms (HABs) in marine and freshwater systems are increasingly common worldwide and are known to cause extensive ecological, economic, and human health problems. In US waters, HABs are encountered in a growing number of locations and are also increasing in duration and severity. This expansion in HABs has led to elevated incidences of poisonous seafood, toxin-contaminated drinking water, mortality of fish and other animals dependent upon aquatic resources (including protected species), public health and economic impacts in coastal and lakeside communities, losses to aquaculture enterprises, and long-term aquatic ecosystem changes. This meeting represented the fourth ACT sponsored workshop that has addressed technology developments for improved monitoring of water-born pathogens and HA species in some form. A primary motivation was to assess the need and community support for an ACT-led Performance Demonstration of Harmful Algae Detection Technologies and Methodologies in order to facilitate their integration into regional ocean observing systems operations. The workshop focused on the identification of region-specific monitoring needs and available technologies and methodologies for detection/quantification of harmful algal species and their toxins along the US marine and freshwater coasts. To address this critical environmental issue, several technologies and methodologies have been, or are being, developed to detect and quantify various harmful algae and their associated toxins in coastal marine and freshwater environments. There are many challenges to nationwide adoption of HAB detection as part of a core monitoring infrastructure: the geographic uniqueness of primary algal species of concern around the country, the variety of HAB impacts, and the need for a clear vision of the operational requirements for monitoring the various species. Nonetheless, it was a consensus of the workshop participants that ACT should support the development of HA detection technology performance demonstrations but that these would need to be tuned regionally to algal species and toxins of concern in order to promote the adoption of state of the art technologies into HAR monitoring networks. [PDF contains 36 pages]

Human Resources Development for Sustainable Aquaculture in Bangladesh

The ennoblement of human resources has become a prime issue in the philosophy of sustainable aquaculture development in the new millennium.Being the planners, designers, conductors and philosophers of sustainable aquaculture, human beings always demand their further improvement at level best from their current positions to bring supreme success in the sector. As sustainable aquaculture is socio-economic –cum-environmental in concept, its operation and management requires constant interplay of various human knowledge for ensuring its smooth direction and for achieving its goal. So, the arrangement of different types and levels of training and education are the great need for the development of personnel involved in sustainable aquaculture route and also for growing awareness of environmental issues. The modus operandi of training and education has to be changed systematically to answer the calls of the needs of the new millennium. In the developing and developed countries where aquaculture plays a vital role in promoting production of aquatic organisms, alleviating of poverty, ensuring environmental compatibility, replenishing and improving the natural stocks, increasing socio-economic upliftment through integrated development approach, developing and managing the aquatic resources, maintaining gene banks and preserving the diversity of fish stocks, it has been already proved that Human resources development (HRD) is inevitable to bring sustainable aquaculture and plays a great role in the flourishment of the system . Different types and levels of training of personnel required for sustainable aquaculture in the new millennium are brought forward in the study. The importance of human resources development (HRD) through specialized training to the personnel is also depicted.

青藏高原东部混农季节性放牧及其对高山林线格局的影响

混农季节性放牧（agropastoral transhumance）通过作物种植和畜牧生产相结合的方式对不同海拔高度带上的资源进行相互补充利用，在亚洲兴都库什地区、青藏高原、横断山、东部及南部非洲、南美安第斯地区等具有悠久的历史。这种传统的生计系统几千年以来一直是居住在该地区的人类社会和自然生态系统相互作用的主要形式之一。这种传统的资源利用方式与山地自然植被以及特殊的山地人类文化和社会特征具有密切的协同演变关系。认识和理解这一关系，是山地生态学和人类学的核心科学问题之一。近年来，山地生态系统的多重功能性及动态演变对山区社会经济可持续发展的重要意义受到人们的不断关注。本文通过对云南省德钦县的12个自然村的混农季节性放牧以及对云南德钦、四川壤塘等山地植被格局特别是高海拔地带植被格局的的详细调查，探讨青藏高原东缘地区混农季节性放牧的主要特征、系统构成及相互关系，及其在全球变化、经济全球化和市场化及现代化过程中的变化趋势，分析混农季节性放牧与高山林线格局及生态系统的互动关系，旨在探讨山地地区人类活动与自然生态系统之间的互动关系，从而为山区社会经济可持续发展、环境建设和生物多样性保护等国家战略提供理论依据。 调查结果表明，混农季节性放牧是一种适应青藏高原东部高山峡谷地区环境因子及自然资源呈明显的垂直分布、资源数量稀少而时空分布异质性极高的生存环境的一种传统经济形式。这种传统的畜牧业的主要生产目的仍然是提供当地基本生存所需的产品，饲养牲口的种类和数量取决于农户的当地需求并且受资源的限制，因而维持在比较低的水平的。分布在不同海拔高度的放牧资源在一年中被牲口利用的时间也不同，互为补充，共同构成混农季节性放牧的资源基础。根据各社区永久居住点的位置和该村的土地资源特别是牧草地资源的分布范围，牲口迁移的距离和格局有较大的差异。。天然牧场仍然是最主要的畜牧业生产资源。混农季节性放牧中的农业系统和牧业系统互为补充，共同构成调查地区完整的的生计系统，农耕活动为放牧活动提供精饲料如粮食等和冬季饲料如秸秆， 其数量往往成为家庭畜牧业生产规模的主要决定因子之一。 通过对牲口数量和结构、牲口的时空迁移格局、牧业活动在整个经济活动中的相对重要性以及牧业活动和作物种植的关系方面的研究分析，混农季节性放牧在近几十年发生了深刻的变化。主要表现在牲口数量总体下降，牲口组成发生变化，牲口移动性降低、牧业活动的经济重要性下降以及牧业活动和种植活动之间的相互依存度降低等。上述变化的根本驱动力是发生在当地、地区及全球尺度上的环境、政治、社会经济、技术和文化等的变化，从而造成当地群众畜牧生产目标、土地利用和劳动力的分布等发生了变化。当地生计系统发生的改变可能会带来对方面而深刻的政治、社会经济、文化和生态影响。 混农季节性放牧这种古老的传统生计策略面临着许多挑战，如冬季饲料短缺、草场退化、缺乏市场竞争力、经济重要性降低、对年轻人缺乏吸引力、国家缺乏专门的政策指导等。与此同时，经济全球化、市场经济、新技术的应用、替代生计机会的增加、国家对于山地生态系统的作用的重新定位等也为传统生计系统转型、实现社会与生态共赢创造了机遇。 混农季节性放牧活动对亚高山及树线交错带生态系统系统的互动方式主要体现在以下几个方面：（1）牲口啃食、践踏等影响森林群落更新，改变森林群落的组成和结构，从而影响森林群落的演替进程和植被格局。林线边缘是搭建夏棚的首选地点，因此林线及树线交错地带就成了牲口活动的主要场所之一；（2）利用火烧开辟、维持和改良高山牧场； 3）在亚高山火灾迹地的放牧活动能够阻止火烧迹地的顺向演替； 4）牧民在林线附近获取建材和薪材等活动影响高山林线附近森林的结构和功能。 在调查区域，梅里雪山、白马雪山、甲午雪山的林线海拔高度在4200-4300m之间； 四川雅江、理塘一线，林线位置多在4300-4400m；四川壤塘二林场一带的林线主体在4100-4200m，在个别地区达到4300m； 在贡嘎山的南坡和东坡一带，林线位置在3600-3700m；而在四川松潘一带，林线位置主体在3700-3800米左右。树线高度的分布趋势和林线一致。混农季节性放牧及其有关人类利用活动使研究地区很多地方高山林线降低、树线交错带宽变窄或消失。在研究地区，总体情况是，阳坡和半阳坡（南坡、西南坡等）的林线和树线比阴坡和半阴坡（北坡、东北坡等）低，变化幅度达20-200m。这种差异主要是为了开辟牧场而人为清除了南向坡自然林线及其以上的植被从而使林线位置下降所致。在南坡自然林线保留得比较好的地方，林线和树线依然可以达到甚至超过北坡林线和树线的高度。放牧活动抑制了高山林线带火烧迹地的天然更新，从而使林线位置保持在目前的位置。 放牧活动对高山林线带森林群落更新的影响是显著的。自然林线内的乔木个体密度特别是新生苗和幼苗的密度大大高于非自然林线。没有放牧的自然林线及树线交错带内的I级个体（新生苗）密度达到725-2917株/公顷，而与之相对的处理样地内I级个体的密度只有0-228株/公顷；II级个体（高度10-50cm）也表现出类似的趋势，在没有放牧的自然林线及树线交错带样方内，其密度达到550-5208株/，而在放牧处理样方内只有14-321株/公顷。在非自然林线带样地内，在有正常放牧的样地内，完全缺乏I级个体。 从相对比例来看，没有放牧的样方内的I、II级个体在全部个体中所占的比例显著高于有放牧活动的样方。放牧使林线交错带的乔木幼苗数量显著减少，从而影响林线及树线交错带森林群落的天然更新过程。林线和树线交错带的灌木对乔木幼苗具有重要的保护作用，能够为树线树种如冷杉等幼苗的定居体提供有利的微气候环境，同时保护苗免受牲口的啃食和践踏。火烧以后接着进行放牧能够100%地抑制高山林线带的幼苗更新。 高山牧场放牧强度降低、使用时间缩短而低海拔地带放牧强度增加是研究地区混农季节性放牧系统的一个显著变化。这种变化也必然会引起各海拔带上的生态系统的变化。放牧强度的降低、生产性用火的停止将导致原来通过人工火烧而降低并通过进一步的火烧和放牧活动来维持的林线及其以上地带的灌木盖度和高度的增加，从而为林线森林群落的扩张创造条件。 青藏高原东部高山峡谷地区是我国重要的山地生态系统，在我国的生物多样性保护、生态环境建设、社会经济可持续发展战略中具有举足轻重的作用。正确认识人类特别是当地传统的生计系统与生态环境系统的互动关系是实现上述战略目标的前提。决策者必须以综合、系统的的视角协调促进社会经济可持续发展、保护生物及文化多样性和维持人、牲口和生态系统之间的平衡的多重目标。 Agropastoral transhumance, which makes a complementary exploitation of the natural resources at different altitudinal belts through a combination of migratory animal husbandry and crop cultivation, has a long history in Hindu-Kush Himalaya, Tibet Plateau, Hengduan Ranges, eastern and southern Africa and the Andes region of south America.For millennia, this traditional livelihood strategy has been one of the main forms of interaction between human societies inhabiting in these regions and their natural ecocystems. A close co-evolutionary relationship has been developed between this indigenous resources management systems and the mountain vegetation systems on the one hand and a unique set of cultural values and social features on the other. Understanding this relationship has been one of the core scientific issues in mountain ecology and anthropology. In recent years, the importance of the multiple functions of the mountain ecosystems and their dynamic changes in the sustainable socio-economic development of the mountain regions has gained increasing attention. This paper, which is based on a detailed study on the agropastoral practices of the 12 natural villages in Deqin County of Yunnan, and the mountainnn vegetation patterns in Deqin of Yunnan and Rangtang County of Sichuan, intends to reveal the major characteristics, system composition and the inter-relations of the subsystems of the agropastoral transhumance in Eastern Tibetan Plateau as well as the trends of changes of the system within the context of global changes, economic globalization and modernity process of China and analyze the relations between agropastoral transhumance and alpine ecosystem, ao as to understand the interactions between human activities and natural ecosystems of the mountains and provide theoretical basis for the national strategies in eocioeconomic development, environmental reconstruction and biodiversity conservation in the mountain regions. Results of the survey indicate that agropastoral transhumance in the investigated area is a traditional economic form that is highly adapted to the eastern Tibet Plateau where the topography features high peaks and deep gorges and where the highly variable environmental parameters and scanty natural resources exhibit a distinct vertical spectrum of distribution and great temporal and spatial heterogeneity. The main objective of pastoral management is still aimed at the production of basic goods and services of local people and thus the type and size of animals raised for each household mainly depend on local needs and are limited by the availability of natural resources. The scale of production is relatively low. Pastoral resources at different altidudinal belts are complementarily used at different seasons of a year and thus form the resources basis for agropastoral production of the study area. Migration distances and patterns vary with the location of the permanent settlements, the elevational distribution range of the resources of the villages concerned. Natural pastures (rangelands) are the main fodder resources and sumplement feedings only account for less than 5% of the total fodder consumption. Crop cultivation and pastoral activities support each other to form a complete livelihood system. The ability of the farmig lands (crop cultivation) to provide the pastoral activities with concentrates and sumplements often becomes a main factor limiting the scale of livestock production at household level. Agropastoral transhumance is experiencing drastic changes in recent decades as is reflected in the size and composition of animals, the seasonal migration pattern, the relative importance of pastoralism in the household economy and the interplays of agricultural and pastoral elements of the system. In general, there is a decline in animal population and mobility, a shift in animal composition to meet new needs arising from changed macro-economic situation, a decrease in the relative importance in the household economy and an increasing decoupling of agro&pastoral relations. The fundamental divers of these changes can be traced to environmental, social, economic, technological and cultural changes from local to global levels and such changes have further caused local changes in livestock management objectives, land use and distribution of labor forces. Changes in local livelihood systems could have profound political, socioeconomic, cultural and ecological conseuquences. Agropastoral transhumance, as an age-old traditional livelihood strategy, is facing multifacet challenges, such as winter fodder shortage, rangeland degradation, lack of market competitiveness, decrease in economic importance, lack of appreciation among the young generation and adequate policies from the government. At the same time, economic globalization, market economy, intrdoctution of new technologies, increase of alternative income generating opportunities and the national re-oreitation of policies on mountain ecosystems have all brought about new opportunities for the transformation of the traditional livelihood system and the synchronized development of local society and the environment. Agropastoral transhumance interacts with the ecosystems at the timberline and treeline ecotone mainly through the following aspects: 1)Animal browsing and stamping affect the regeneration process of the forest communities and alters the composition and structure of the forest which in turn affect the succession process and vegetation pattern of the forest communities. Forest edges are the priority locations for summer houses and therefore the timeline and treeline area becomes the major venues of aninal activities; （2）herders create, maintain and improve pastures through burning that remove the forest communities at the timeline and treeline ecotone; 3）immediate grazing on the fire sites can significantly prevent the fire sites from perogressive succession; and 4）herders harvesting of construction timber and firewoods affects the structure and functions of the forest communities at the timberline and treeline zone. Timberline position in the survey region shows geographical variations. It is around 4200-4300m in Meilixueshan, Baimaxueshan and Jiawuxueshan in Northwest of Yunnan and rises to 4300-4400m in Yajiang County and Litang County of Sichuan. In Rangtang of Sichuan, it is between 4100-4200m, though reaching 4300m in localized sites. In the southern and eastern slopes of Gongga Mountain, the timberline is only between 3600m and 3700m and in Songpan County at the upper reach of the Minjiang River the timberline is around 3700-3800m.Treeline pattern follows similar trend. In many places, agropastoral transhumance and related human activities have lowered the timberline and treeline and narrowed or removed the treeline ecotone. In the area of survey, generally speaking, timberlines and treelines are lower on the southern slopes than on the northern slopes, with a difference between 20 and 200m. This is mainly because that the use of fires to crerate pastures has removed the forest vegetation at the previous timberline and above. In fact, in many places, well-preserved forests on the south slopes have even high timberline position that the corresponding northern slopes. At subalpine zone, grazing activities could have prohibited the natural regeneration of many forest fire sites and maintained the forest position at the present level. Grazing has a significant impact on the regernation process of forest communities at the timberline zone. Natural timberline and treeline ecotone has much higher density of treeline species individuals especially the emergents and seedlings than the timberlines that are maintained by human activities. In natural timberline and treelien ecotone without grazing interference, the density of the I Class seedlings (less than 10cm in height) ranges 725-2917 /hm2; while that in the treatment plots (with grazing disturbance) is only 0-228//hm2；II Class seedlings (10-50cm）exhibit similar density trends, reaching 550-5208//hm2 in natural timberline without grazing but only 14-321//hm2 in the plots with grazing treatment. In the man-created timberlines, there is no I Class seedling at all in plots with normal grazing activities. In relative terms, in plots without grazing activities, the propotion of I Class and II Class seedlings is much higher than that in plots with grazing. Grazing activities have significantly reduced the number of seedlings in the timberline ane treeline ecotone, and thus affect the natural regeneration process of the forests. Shrubs at the timberline and treeline ecotone can effectively protect the seedlings from severe climate and animal tramping, thus increasing the survival rate of the seedlings. Grazing following fires can completely inhibit forest regeneration process at timberline. Changes in agropastoral transhumance will have great impact on the timberline and treeline pattern of the studied area. The decrease in grazing intensity on alpine pastrues and the cessation of the use of fires will result an increase in the cover and height of shrubs above the present human-maintained treeline, which will create further condition for the expansion of timberline forest communities. Eastern Tibet Plateau harbors some most important mountain ecosystems of China that are of vital importance to the country’s strategy in biodiversity conservation, environmental construction and sustainable sociaoeconomic development. A proper knowledge of the interactions between traditional livelihood systems and the ecosystems in the region is a precondition to the realization of the above strategic goals. Therefore, the decision-makers must have a holistic and systemic perspective so as to integrate the multiple objectives of promoting sustainable socioeconomic development, conserving biological and cultural diversity and maintaining the balances among people, animal population and the ecosystems.

Enhancement of atmospheric pressure chemical ionization for the determination of free and glycine-conjugated bile acids in human serum

A highly sensitive and accurate method based on the precolumn derivatization of bile acids (BA) with a high ionization efficiency labeling reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl-benzenesulfonate (BDEBS) coupled with LC/MS has been developed. After derivatization, BA molecules introduced a weak basic nitrogen atom into the molecular core structure that was readily ionized in commonly used acidic HPLC mobile phases. Derivatives were sufficiently stable to be efficiently analyzed by atmospheric pressure chemical ionization (APCI)-MS/MS in positive-ion mode. The MS/MS spectra of BA derivatives showed an intense protonated molecular ion at m/z [M + H](+). The collision-induced dissociation of the molecular ion produced fragment ions at [MH - H2O](+), [MH - 2H(2)O](+), [MH - 3H(2)O](+). The characteristic fragment ions were at m/z 320.8, 262.8, and 243.7 corresponding to a cleavage of N - CO, O - CO, and C - OCC, respectively, and bonds of derivatized molecules. The selected reaction monitoring, based on the m/z [M + H]+ -> [MH - H2O](+), [MH - H2O](+), [MH - 2H(2)O](+), [MH-3H(2)O](+), 320.8, 262.8, and 243.7 transitions, was highly specific for the BA derivatives. The LODs for APCI in a positive-ion mode, at an S/N of 5, were 44.36-153.6 fmol. The validation results showed high accuracy in the range of 93-107% and the mean interday precision for all standards was < 15% at broad linear dynamic ranges (0.0244-25nmol/mL). Good linear responses were observed with coefficients of > 0.9935 in APCI/MS detection. Therefore, the facile BDEBS derivatization coupled with mass spectrometric analysis allowed the development of a highly sensitive and specific method for the quantitation of trace levels of the free and glycine-conjugated BA from human serum samples.

Age- and activity-related differences in the abundance of Myosin essential and regulatory light chains in human muscle

Traditional methods for phenotyping skeletal muscle (e.g., immunohistochemistry) are labor-intensive and ill-suited to multixplex analysis, i.e., assays must be performed in a series. Addressing these concerns represents a largely unmet research need but more comprehensive parallel analysis of myofibrillar proteins could advance knowledge regarding age- and activity-dependent changes in human muscle. We report a label-free, semi-automated and time efficient LC-MS proteomic workflow for phenotyping the myofibrillar proteome. Application of this workflow in old and young as well as trained and untrained human skeletal muscle yielded several novel observations that were subsequently verified by multiple reaction monitoring (MRM).We report novel data demonstrating that human ageing is associated with lesser myosin light chain 1 content and greater myosin light chain 3 content, consistent with an age-related reduction in type II muscle fibers. We also disambiguate conflicting data regarding myosin regulatory light chain, revealing that age-related changes in this protein more closely reflect physical activity status than ageing per se. This finding reinforces the need to control for physical activity levels when investigating the natural process of ageing. Taken together, our data confirm and extend knowledge regarding age- and activity-related phenotypes. In addition, the MRM transitions described here provide a methodological platform that can be fine-tuned to suite multiple research needs and thus advance myofibrillar phenotyping.

Chondrogenesis and mineralization during in vitro culture of human mesenchymal stem cells on three-dimensional woven scaffolds.

Human mesenchymal stem cells (hMSCs) and three-dimensional (3D) woven poly(ɛ-caprolactone) (PCL) scaffolds are promising tools for skeletal tissue engineering. We hypothesized that in vitro culture duration and medium additives can individually and interactively influence the structure, composition, mechanical, and molecular properties of engineered tissues based on hMSCs and 3D poly(ɛ-caprolactone). Bone marrow hMSCs were suspended in collagen gel, seeded on scaffolds, and cultured for 1, 21, or 45 days under chondrogenic and/or osteogenic conditions. Structure, composition, biomechanics, and gene expression were analyzed. In chondrogenic medium, cartilaginous tissue formed by day 21, and hypertrophic mineralization was observed in the newly formed extracellular matrix at the interface with underlying scaffold by day 45. Glycosaminoglycan, hydroxyproline, and calcium contents, and alkaline phosphatase activity depended on culture duration and medium additives, with significant interactive effects (all p < 0.0001). The 45-day constructs exhibited mechanical properties on the order of magnitude of native articular cartilage (aggregate, Young's, and shear moduli of 0.15, 0.12, and 0.033 MPa, respectively). Gene expression was characteristic of chondrogenesis and endochondral bone formation, with sequential regulation of Sox-9, collagen type II, aggrecan, core binding factor alpha 1 (Cbfα1)/Runx2, bone sialoprotein, bone morphogenetic protein-2, and osteocalcin. In contrast, osteogenic medium produced limited osteogenesis. Long-term culture of hMSC on 3D scaffolds resulted in chondrogenesis and regional mineralization at the interface between soft, newly formed engineered cartilage, and stiffer underlying scaffold. These findings merit consideration when developing grafts for osteochondral defect repair.

The evolutionary origins of human patience: temporal preferences in chimpanzees, bonobos, and human adults.

To make adaptive choices, individuals must sometimes exhibit patience, forgoing immediate benefits to acquire more valuable future rewards [1-3]. Although humans account for future consequences when making temporal decisions [4], many animal species wait only a few seconds for delayed benefits [5-10]. Current research thus suggests a phylogenetic gap between patient humans and impulsive, present-oriented animals [9, 11], a distinction with implications for our understanding of economic decision making [12] and the origins of human cooperation [13]. On the basis of a series of experimental results, we reject this conclusion. First, bonobos (Pan paniscus) and chimpanzees (Pan troglodytes) exhibit a degree of patience not seen in other animals tested thus far. Second, humans are less willing to wait for food rewards than are chimpanzees. Third, humans are more willing to wait for monetary rewards than for food, and show the highest degree of patience only in response to decisions about money involving low opportunity costs. These findings suggest that core components of the capacity for future-oriented decisions evolved before the human lineage diverged from apes. Moreover, the different levels of patience that humans exhibit might be driven by fundamental differences in the mechanisms representing biological versus abstract rewards.

Human-chimpanzee differences in a FZD8 enhancer alter cell-cycle dynamics in the developing neocortex.

The human neocortex differs from that of other great apes in several notable regards, including altered cell cycle, prolonged corticogenesis, and increased size [1-5]. Although these evolutionary changes most likely contributed to the origin of distinctively human cognitive faculties, their genetic basis remains almost entirely unknown. Highly conserved non-coding regions showing rapid sequence changes along the human lineage are candidate loci for the development and evolution of uniquely human traits. Several studies have identified human-accelerated enhancers [6-14], but none have linked an expression difference to a specific organismal trait. Here we report the discovery of a human-accelerated regulatory enhancer (HARE5) of FZD8, a receptor of the Wnt pathway implicated in brain development and size [15, 16]. Using transgenic mice, we demonstrate dramatic differences in human and chimpanzee HARE5 activity, with human HARE5 driving early and robust expression at the onset of corticogenesis. Similar to HARE5 activity, FZD8 is expressed in neural progenitors of the developing neocortex [17-19]. Chromosome conformation capture assays reveal that HARE5 physically and specifically contacts the core Fzd8 promoter in the mouse embryonic neocortex. To assess the phenotypic consequences of HARE5 activity, we generated transgenic mice in which Fzd8 expression is under control of orthologous enhancers (Pt-HARE5::Fzd8 and Hs-HARE5::Fzd8). In comparison to Pt-HARE5::Fzd8, Hs-HARE5::Fzd8 mice showed marked acceleration of neural progenitor cell cycle and increased brain size. Changes in HARE5 function unique to humans thus alter the cell-cycle dynamics of a critical population of stem cells during corticogenesis and may underlie some distinctive anatomical features of the human brain.

Core and Shell Song Systems Unique to the Parrot Brain.

The ability to imitate complex sounds is rare, and among birds has been found only in parrots, songbirds, and hummingbirds. Parrots exhibit the most advanced vocal mimicry among non-human animals. A few studies have noted differences in connectivity, brain position and shape in the vocal learning systems of parrots relative to songbirds and hummingbirds. However, only one parrot species, the budgerigar, has been examined and no differences in the presence of song system structures were found with other avian vocal learners. Motivated by questions of whether there are important differences in the vocal systems of parrots relative to other vocal learners, we used specialized constitutive gene expression, singing-driven gene expression, and neural connectivity tracing experiments to further characterize the song system of budgerigars and/or other parrots. We found that the parrot brain uniquely contains a song system within a song system. The parrot "core" song system is similar to the song systems of songbirds and hummingbirds, whereas the "shell" song system is unique to parrots. The core with only rudimentary shell regions were found in the New Zealand kea, representing one of the only living species at a basal divergence with all other parrots, implying that parrots evolved vocal learning systems at least 29 million years ago. Relative size differences in the core and shell regions occur among species, which we suggest could be related to species differences in vocal and cognitive abilities.

The NRF-1/α-PAL transcription factor regulates human E2F6 promoter activity

E2F6 is widely expressed in human tissues and cell lines. Recent studies have demonstrated its involvement in developmental patterning and in the regulation of various genes implicated in chromatin remodelling. Despite a growing number of studies, nothing is really known concerning the E2F6 expression regulation. To understand how cells control E2F6 expression, we analysed the activity of the previously cloned promoter region of the human E2F6 gene. DNase I footprinting, gel electrophoretic-mobility shift, transient transfection and site-directed mutagenesis experiments allowed the identification of two functional NRF-1/α-PAL (nuclear respiratory factor-1/α-palindrome-binding protein)-binding sites within the human E2F6 core promoter region, which are conserved in the mouse and rat E2F6 promoter region. Moreover, ChIP (chromatin immunoprecipitation) analysis demonstrated that overexpressed NRF-1/α-PAL is associated in vivo with the E2F6 promoter. Furthermore, overexpression of full-length NRF-1/α-PAL enhanced E2F6 promoter activity, whereas expression of its dominant-negative form reduced the promoter activity. Our results indicate that NRF-1/α-PAL is implicated in the regulation of basal E2F6 gene expression.

15N NMR relaxation data reveal significant chemical exchange broadening in the alpha-domain of human α-lactalbumin

Human alpha-lactalbumin (alpha-LA), a 123-residue calcium-binding protein, has been studied using (15)N NMR relaxation methods in order to characterize backbone dynamics of the native state at the level of individual residues. Relaxation data were collected at three magnetic field strengths and analyzed using the model-free formalism of Lipari and Szabo. The order parameters derived from this analysis are generally high, indicating a rigid backbone. A total of 46 residues required an exchange contribution to T(2); 43 of these residues are located in the alpha-domain of the protein. The largest exchange contributions are observed in the A-, B-, D-, and C-terminal 3(10)-helices of the alpha-domain; these residues have been shown previously to form a highly stable core in the alpha-LA molten globule. The observed exchange broadening, along with previous hydrogen/deuterium amide exchange data, suggests that this part of the alpha-domain may undergo a local structural transition between the well-ordered native structure and a less-ordered molten-globule-like structure.