A light and electron microscopic examination of Ehrlichia sennetsu in cultured human endothelial cells

Umbilical cord-derived human endothelial cells were used for the propagation of Ehrlichia sennetsu. Scanning electron microscopic studies revealed numerous pleomorphic ehrlichiae extending from the surface of infected human endothelial cells. Transmission electron microscopic examination of the cells revealed E. sennetsu-induced cytopathic changes at the ultrastructural level. Ehrlichiae of variable size, shape and density were located individually and in clusters enclosed within membrane lined vacuoles. The cultured human endothelial cell provides an optimal environment for the growth of E. sennetsu and is a suitable in vitro model for the study of the cytopathic effects of this human pathogen.     

Transfer of cultured rabbit embryos

Embryo transfer experiments were carried out to study the developmental capacity of cultured rabbit embryos when transferred to recipients of variable postovulatory maturity. Rabbit embryos were flushed from the oviduct at 26 hours postcoitum (pc) and cultured in a modified Ham's F-10 medium supplemented with bovine serum albumin (BSA) for a period of 70 hours. At 96 hours pc the cultured embryos, which ranged from the early morula to the expanding blastocyst stage, were transferred to pseudopregnant recipients mated to vasectomized males 36 to 96 hours prior to the transfer procedure. Greatest embryo survival occurred when transfers were made to either the oviducts or uterine horns of recipients at 48 hours pc. Intermediate results for both implantation rates and number of young born were obtained with recipients at 36, 60, 72, and 84 hours pc. Transferred embryos consistently failed to survive the uterine environment of recipients 96 hours pc at transfer although this group was synchronous with embryonic chronological age. Oviductal transfers were generally more successful than uterine transfers. Markedly higher rates of embryo survival resulted from embryos that were collected 60 and 72 hours pc and transferred directly to synchronous recipients without an interim period of culture. Dissimilarity of development for in vivo grown rabbit embryos and those cultured in synthetic medium is demonstrated.     

A stochastic model of a temperature-dependent population

The theoretical basis is developed for a population model which allows the use of constant temperature experimental data in predicting the size of an insect population for any variable temperature environment. The model is based on a stochastic analysis of an insect's mortality, development, and reproduction response to temperature. The key concept in the model is the utilization of a physiological time scale. Different temperatures affect the population by increasing an individual's physiological age by differing rates. Conditions for the temperature response properties are given which establish the validity of the model for variable temperature regimes. These conditions refer to the relationship between chronological and physiological age. Reasonable agreement between the model and field populations demonstrates the practicality of this approach.     

Inference of Cross-Level Interaction between Genes and Contextual Factors in a Matched Case-Control Metabolic Syndrome Study: A Bayesian Approach

Genes, environment, and the interaction between them are each known to play an important role in the risk for developing complex diseases such as metabolic syndrome. For environmental factors, most studies focused on the measurements observed at the individual level, and therefore can only consider the gene-environment interaction at the same individual scale. Indeed the group-level (called contextual) environmental variables, such as community factors and the degree of local area development, may modify the genetic effect as well. To examine such cross-level interaction between genes and contextual factors, a flexible statistical model quantifying the variability of the genetic effects across different categories of the contextual variable is in need. With a Bayesian generalized linear mixed-effects model with an unconditional likelihood, we investigate whether the individual genetic effect is modified by the group-level residential environment factor in a matched case-control metabolic syndrome study. Such cross-level interaction is evaluated by examining the heterogeneity in allelic effects under various contextual categories, based on posterior samples from Markov chain Monte Carlo methods. The Bayesian analysis indicates that the effect of rs1801282 on metabolic syndrome development is modified by the contextual environmental factor. That is, even among individuals with the same genetic component of PPARG_Pro12Ala, living in a residential area with low availability of exercise facilities may result in higher risk. The modification of the group-level environment factors on the individual genetic attributes can be essential, and this Bayesian model is able to provide a quantitative assessment for such cross-level interaction. The Bayesian inference based on the full likelihood is flexible with any phenotype, and easy to implement computationally. This model has a wide applicability and may help unravel the complexity in development of complex diseases.     

基于可变模糊识别模型的海水环境质量评价

海水环境质量评价可以视为是一个具有确定性的评价指标和评价标准与具有不确定性的评价因子及其含量变化相结合的分析过程,各评价指标含量具有中介过渡性,构建了基于可变模糊集合理论的海水环境质量评价方法(简称“VFEM”),并以莱州湾海水水质为研究对象,应用提出的可变模糊评价模型对该区域海水水质状况进行评价,实例应用表明,该模型通过可变模型参数变化(a,p),以线性模型与非线性模型相结合,将最后稳定结果作为海洋水质环境的最后评价结果,从而确定水质评价等级,评价结果更为可信,同时该模型通过级别特征值精确区分各水质采样点的水质优劣,对各水质采样点的水质级别有更准确的定位,为海水环境质量综合评价提供了一种合理而适用的方法.     

Coupling of transcription termination to RNAi

Microbes require multiple essential elements that they acquire from the environment independently. Here we investigate how microbial stoichiometry and uptake rates depend on the conditions in which they grow. We modify a recent model of growth based on a multinutrient extension of the Droop model to allow a trade-off between ability to acquire two essential resources. In a static analysis, we show that the optimal allocation strategy is the one that results in co-limitation by both nutrients. We then add a dynamic equation to model the physiological acclimation uptake rates in changing conditions. This dynamic model predicts that the response of organismal stoichiometry to nutrient supply ratio can vary over time. The response of organismal stoichiometry and growth rate to a nutrient pulse depends on the speed at which cells adapt their uptake rates. In a variable environment, very fast or very slow acclimation may be better strategies than intermediate speed acclimation. We suggest experimental tests of the model and avenues for future model development.     

Optimal control strategy of malaria vector using genetically modified mosquitoes

The development of transgenic mosquitoes that are resistant to diseases may provide a new and effective weapon of diseases control. Such an approach relies on transgenic mosquitoes being able to survive and compete with wild-type populations. These transgenic mosquitoes carry a specific code that inhibits the plasmodium evolution in its organism. It is said that this characteristic is hereditary and consequently the disease fades away after some time. Once transgenic mosquitoes are released, interactions between the two populations and inter-specific mating between the two types of mosquitoes take place. We present a mathematical model that considers the generation overlapping and variable environment factors. Based on this continuous model, the malaria vector control is formulated and solved as an optimal control problem, indicating how genetically modified mosquitoes should be introduced in the environment. Numerical simulations show the effectiveness of the proposed control.     

Apple Fruit Diameter and Length Estimation by Using the Thermal and Sunshine Hours Approach and Its Application to the Digital Orchard Management Information System

In apple cultivation, simulation models may be used to monitor fruit size during the growth and development process to predict production levels and to optimize fruit quality. Here, Fuji apples cultivated in spindle-type systems were used as the model crop. Apple size was measured during the growing period at an interval of about 20 days after full bloom, with three weather stations being used to collect orchard temperature and solar radiation data at different sites. Furthermore, a 2-year dataset (2011 and 2012) of apple fruit size measurements were integrated according to the weather station deployment sites, in addition to the top two most important environment factors, thermal and sunshine hours, into the model. The apple fruit diameter and length were simulated using physiological development time (PDT), an indicator that combines important environment factors, such as temperature and photoperiod, as the driving variable. Compared to the model of calendar-based development time (CDT), an indicator counting the days that elapse after full bloom, we confirmed that the PDT model improved the estimation accuracy to within 0.2 cm for fruit diameter and 0.1 cm for fruit length in independent years using a similar data collection method in 2013. The PDT model was implemented to realize a web-based management information system for a digital orchard, and the digital system had been applied in Shandong Province, China since 2013. This system may be used to compute the dynamic curve of apple fruit size based on data obtained from a nearby weather station. This system may provide an important decision support for farmers using the website and short message service to optimize crop production and, hence, economic benefit.     

Habitat selection in a variable environment

A Monte Carlo simulation scheme was utilized to determine optimal strategies of habitat utilization in a variable environment. The model allows for differences in quality among habitats at any one time and for varying levels of environmental variance and autocorrelation. When habitats are on the average equal in quality, tracking of temporal fluctuations in environment through variable habitat selection is universally advantageous with the gain in fitness limited by environmental variance, autocorrelation, and number of available habitats. Average differences in quality among habitats will restrict the advantage of variable habitat utilization (over complete usage of the average better habitat) to cases of high environmental autocorrelation or high ratios of enviromental variance to mean habitat separation. Extending an earlier prediction of Levins (1965), the average heterozygosity per individual in a natural population should increase with increasing environmental variance.     

Effect of substrate stiffness on early mouse embryo development

It is becoming increasingly clear that cells are remarkably sensitive to the biophysical cues of their microenvironment and that these cues play a significant role in influencing their behaviors. In this study, we investigated whether the early pre-implantation embryo is sensitive to mechanical cues, i.e. the elasticity of the culture environment. To test this, we have developed a new embryo culture system where the mechanical properties of the embryonic environment can be precisely defined. The contemporary standard environment for embryo culture is the polystyrene petri dish (PD), which has a stiffness (1 GPa) that is six orders of magnitude greater than the uterine epithelium (1 kPa). To approximate more closely the mechanical aspects of the in vivo uterine environment we used polydimethyl-siloxane (PDMS) or fabricated 3D type I collagen gels (1 kPa stiffness, Col-1k group). Mouse embryo development on alternate substrates was compared to that seen on the petri dish; percent development, hatching frequency, and cell number were observed. Our results indicated that embryos are sensitive to the mechanical environment on which they are cultured. Embryos cultured on Col-1k showed a significantly greater frequency of development to 2-cell (68±15% vs. 59±18%), blastocyst (64±9.1% vs. 50±18%) and hatching blastocyst stages (54±25% vs. 21±16%) and an increase in the number of trophectodermal cell (TE,65±13 vs. 49±12 cells) compared to control embryos cultured in PD (mean±S.D.; p<.01). Embryos cultured on Col-1k and PD were transferred to recipient females and observed on embryonic day 12.5. Both groups had the same number of fetuses, however the placentas of the Col-1k fetuses were larger than controls, suggesting a continued effect of the preimplantation environment. In summary, characteristics of the preimplantation microenvironment affect pre- and post-implantation growth.     

Prototype of a novel autonomous perfusion chamber for long-term culturing and in situ investigation of various cell types.

In the context of a neurobionic approach to chemical analysis and sensorics, this article depicts the development of a miniaturized autonomous perfusion chamber setup for the growth and the electrical as well as optical investigation of (neural) cell cultures in vitro. We suggest an autonomous, modular, temperature-controlled, transparent, and sealed perfusion cell culture housing adaptable to various mounts, sizes and different needs. The design includes the electronics of a temperature and medium supply control unit. The setup combines the possibility of uninterrupted cell culturing with simultaneous microscopic and analytical investigation of variable amounts of cells or organs of human, animal, or plant origin under sterile conditions on different substrates without the need of an external incubator or a sterile working environment. Its use is demonstrated exemplarily with neuronal cultures from embryonic chicken that were cultured in a prototype system for 3 weeks. It turned out that cell survival in such a chamber was prolonged with timed medium flow rather than continuous perfusion.     

Impaired maturation, fertilization, and embryonic development of porcine oocytes following exposure to an environmentally relevant organochlorine mixture

The reproductive health risks related to exposure to persistent organic pollutants in the environment remain controversial. This debate is partly because most studies have investigated only one or two chemicals at a time, whereas populations are exposed to a large spectrum of persistent chemicals in their environment. Using the pig as a toxicological model, we hypothesized that exposing immature cumulus-oocyte complexes to an organochlorine mixture during in vitro maturation (IVM) would adversely affect oocyte maturation, fertilization, and subsequent embryo development. This organochlorine mixture mimics that which contaminates the Arctic marine food chain. Cumulus-oocyte complexes were cultured in IVM medium containing increasing concentrations of the organochlorine mixture, similar to that found in women of highly exposed populations. Organochlorines reduced the quality of cumulus expansion and the viability of cumulus cells in a dose-response manner. The proportion of apoptotic cumulus cells also increased due to organochlorine exposure. Half of the oocytes were fixed after insemination, and the remainders were cultured for 8 days. Concentrations of organochlorines did not affect the rates of oocyte degeneration, sperm penetration, and development to morula. However, incidence of incompletely matured oocytes increased and polyspermy rate decreased, both in a dose-response manner with increasing organochlorine concentrations. Blastocyst formation and number of cells per blastocyst declined with organochlorine concentration. Exposing porcine cumulus-oocyte complexes to an environmentally pertinent organochlorine mixture during IVM disturbs oocyte development, supporting recent concerns that such pollutants harm reproductive health in humans and other mammalian species.     

Environments and evolution: interactions between stress, resource inadequacy and energetic efficiency

Evolutionary change is interpreted in terms of the near-universal ecological scenario of stressful environments. Consequently, there is a premium on the energetically efficient exploitation of resources in a resource-inadequate world. Under this environmental model, fitness can be approximated to energetic efficiency especially towards the limits of survival. Furthermore, fitness at one stage of the life-cycle should correlate with fitness at other stages, especially for development time, survival and longevity; 'good genotypes' under stress should therefore be at a premium. Conservation in the wild depends primarily on adaptation to abiotically changing habitats since towards the limits of survival, genomic variation is rarely restrictive. The balance between energetic costs under variable environments and energy from resources provides a model for interpreting evolutionary stasis, punctuational and gradual change, and specialist diversification. Ultimately, a species should be in an equilibrium between the physiology of an organism and its adaptation to the environment. The primary key to understanding evolutionary change should therefore be ecological, highlighting energy availability in a stressed world; this approach is predictive for various patterns of evolutionary change in the living and fossil biota.     

环渤海沿海地区土地承载力时空分异特征

沿海地区是中国未来社会经济发展的重点区域,近年来沿海地区人地矛盾越来越突出,研究沿海地区土地承载力具有重要意义。针对以往评价中的不足,采用韦伯-费希纳定律确定指标评价标准;利用D-S证据理论进行主客观综合权重的计算,运用可变模糊集理论及模型对土地承载力进行综合评价,并以环渤海沿海地区进行实例分析。研究结果表明,2000—2011年,环渤海地区土地承载力整体呈持续上升趋势,但空间分布差异显著,天津、唐山、大连、盘锦、青岛、烟台、东营、威海土地承载力相对较高,锦州、营口、秦皇岛、滨州、潍坊、日照属于中等水平,沧州、丹东、葫芦岛四市土地承载力相对较低。可变模糊评价模型应用于土地承载力评价,计算简便,可操作性强,评价结果可信度高。     

温室甜瓜营养生长期日蒸腾量估算模型

建立了基于温室环境参数、甜瓜生长发育参数和土壤水分参数的温室甜瓜日蒸腾量估算模型,以研究温室条件下甜瓜蒸腾量的估算方法.根据温室内特定环境对Penman-Monteith方程中空气动力项进行修正,推导出了适于计算温室条件下参考作物蒸腾量的温室环境因子子模型;以甜瓜叶面积指数为自变量构建了作物因子子模型,模型形式为线性函数;以土壤相对有效含水量为自变量构建了土壤水分因子子模型,模型形式为对数函数.采用分期播种法,根据周年不同播期实测数据对模型参数进行估计和分析.采用土壤相对含水量分别为80%、70%、60%的实测蒸腾数据,对模型在充分灌溉和节水灌溉条件下的预测精度进行了检验,模拟值的平均相对误差分别为11.5%、16.2%、16.9%.所建蒸腾模型是对Penman-Monteith公式在温室环境和节水灌溉条件下的有益探索,具有重要推广应用价值.     

GALGO: an R package for multivariate variable selection using genetic algorithms

SUMMARY: The development of statistical models linking the molecular state of a cell to its physiology is one of the most important tasks in the analysis of Functional Genomics data. Because of the large number of variables measured a comprehensive evaluation of variable subsets cannot be performed with available computational resources. It follows that an efficient variable selection strategy is required. However, although software packages for performing univariate variable selection are available, a comprehensive software environment to develop and evaluate multivariate statistical models using a multivariate variable selection strategy is still needed. In order to address this issue, we developed GALGO, an R package based on a genetic algorithm variable selection strategy, primarily designed to develop statistical models from large-scale datasets.     

动物细胞培养技术在人造肉研究中的应用

人造肉作为2018年全球十大突破和新兴科技之一,因其来源可追溯、食品安全性和绿色可持续等优势得到广泛的关注。欧美等国家已经投入大量资源开展细胞培养人造肉研究,未来将对我国的肉制品及食品市场造成一定的冲击。现阶段,细胞培养人造肉生产的挑战在于如何高效模拟动物肌肉组织生长环境,并在生物反应器中实现大规模的生产。尽管动物细胞组织培养技术已经得到深入的研究,并取得了不同程度的成功应用,但由于现有动物细胞组织培养成本与技术要求较高,仍不能实现大规模的产业化培养。因此,对于人造肉的生产来说,开发高效、安全的大规模细胞培养技术是亟需解决的问题,可以有效降低生产成本,实现产业化应用。文中通过介绍基于人造肉生物制造的动物细胞组织培养技术研究现状,具体阐述了目前的挑战和关键技术问题,并初步探讨了其可能的解决策略和应用前景。     

Oocyte Ca2+ spike acquisition during in vitro development of early preantral follicles: influence of age and hormonal supplementation

Calcium signalling is involved in important events in oocytes, such as meiotic competence acquisition. We have previously demonstrated the positive influence of animal age and gonadotropin stimulation in vivo regarding the ability of oocytes recovered from preantral follicles to exhibit calcium spikes. In the present work we determined whether preantral follicle development in vitro also allows oocytes to acquire calcium signalling activity. We also aimed to verify the influence of animal age, FSH + LH and/or insulin on oocyte calcium spike acquisition during preantral follicle culture. Early preantral follicles were isolated from 12-day-old and 1- to 3-month-old F1 hybrid mice and cultured individually for either 2 or 6 days. At the end of the culture period the oocytes were processed for calcium imaging by confocal microscopy. We show that oocytes recovered from cultured preantral follicles exhibit variable calcium spike activity rates, depending on animal age, culture duration and hormonal supplementation. Oocytes recovered from adult animals continue to exhibit calcium spikes, and those recovered from juveniles acquire that activity after culture. Insulin and gonadotropins in combination account for an early and maintained inhibitory effect on calcium signalling acquisition by oocytes. Insulin alone also leads to an early inhibitory effect, which, however, disappears with longer culture periods. Contrary to the complex in vivo situation, the acquisition of calcium signalling by oocytes in a controlled in vitro environment does not seem to be dependent on gonadotropins alone.     

Tissue engineering: generation of differentiated artificial tissues for biomedical applications

A new field in biomedical science has been established. Cell biologists, engineers, and surgeons now work within a team. Artificial connective, epithelial, or neuronal tissues are being constructed using living cells and different kinds of biomaterials. Numerous companies and laboratories are presenting dynamic developments in this field. Prognoses predict that, at the beginning of the coming century, the industry of tissue engineering will reach the importance of the present genetic technology. An enormous demand for organ and tissue transplants motivates research activities and drives the acquisition of innovative techniques and creative solutions. At the front of this development is the creation of artificial skin for severely burned patients and the generation of artificial cartilage for implantation in articular joint diseases. Future challenges are the construction of liver organoids and the development of an artificial kidney on the basis of cultured cells. In this paper we show strategies, needs, tools, and equipment for tissue engineering. The presupposition for all projects is the induction, development, and maintenance of differentiation within the tissue under in vitro conditions. As experiments in conventional culture dishes continued to fail, new cell and tissue culture methods had to be developed. Tissues are cultured under conditions as close as possible to their natural environment. To optimize adherence or embedding, cells are grown on novel tissue carriers and on individually selected biomatrices or scaffolds. The tissues are subsequently transferred into different types of containers for permanent perfusion with fresh culture medium. This guarantees constant nutrition of the developing tissue and prevents the accumulation of harmful metabolites. An organo-typical environment for epithelial cells, for example, is obtained in gradient containers, which are permanently superfused at the apical and basal sides with different media. Long term experiments result in cultured tissues in a quality thus far unreached.     

Abiotic and biotic determinants of the seasonal dynamics of the tick Rhipicephalus appendiculatus in South Africa

Abstract. The creation of a generic population model for the tick Rhipicephalus appendiculatus requires a detailed, quantified understanding of the interactions of these ticks with their biotic and abiotic environment in the different parts of their range, from the tropical regions of East Africa to the temperate regions of South Africa. The much greater seasonal variation in climatic conditions, particularly temperature, further from the equator introduces variable development rates and diapause into the life cycle. Estimates of natural temperature-dependent interstadial development periods, derived from a combination of published laboratory and field data, were applied to published data on the seasonal abundance of R.appendiculatus on three farms in the Eastern Cape Province of South Africa. This enabled an assessment of which ticks of one stage give rise to which ticks of the next stage, from which (a) the onset and duration of diapause in unfed adults, and (b) seasonal interstadial mortality indices in the form of fc-values, could be estimated. The contribution of biotic (tick density) and abiotic (climatic) factors as predictors of mortality at each life stage was investigated by step-wise multiple regression. Density-independent mortality at the female-to-larval stage is correlated with geographically variable climatic factors, minimum temperature at two farms and minimum relative humidity at the third. The other two stages are governed by density-dependent mortality, which, it is argued, may be caused largely by the hosts' acquired resistance to ticks. As expected on theoretical grounds, this density dependence is weaker nearer to the edge of the tick's range and for the more vulnerable immature stages.