An Ex vivo Culture System to Study Thyroid Development

The thyroid is a bilobated endocrine gland localized at the base of the neck, producing the thyroid hormones T3, T4, and calcitonin. T3 and T4 are produced by differentiated thyrocytes, organized in closed spheres called follicles, while calcitonin is synthesized by C-cells, interspersed in between the follicles and a dense network of blood capillaries. Although adult thyroid architecture and functions have been extensively described and studied, the formation of the “angio-follicular” units, the distribution of C-cells in the parenchyma and the paracrine communications between epithelial and endothelial cells is far from being understood.This method describes the sequential steps of mouse embryonic thyroid anlagen dissection and its culture on semiporous filters or on microscopy plastic slides. Within a period of four days, this culture system faithfully recapitulates in vivo thyroid development. Indeed, (i) bilobation of the organ occurs (for e12.5 explants), (ii) thyrocytes precursors organize into follicles and polarize, (iii) thyrocytes and C-cells differentiate, and (iv) endothelial cells present in the microdissected tissue proliferate, migrate into the thyroid lobes, and closely associate with the epithelial cells, as they do in vivo.Thyroid tissues can be obtained from wild type, knockout or fluorescent transgenic embryos. Moreover, explants culture can be manipulated by addition of inhibitors, blocking antibodies, growth factors, or even cells or conditioned medium. Ex vivo development can be analyzed in real-time, or at any time of the culture by immunostaining and RT-qPCR.In conclusion, thyroid explant culture combined with downstream whole-mount or on sections imaging and gene expression profiling provides a powerful system for manipulating and studying morphogenetic and differentiation events of thyroid organogenesis.     

Both facilitation and limiting similarity shape the species coexistence in dry alkali grasslands

Facilitation is an important driver of community assembly, and often overwhelms the effect of competition in stressed habitats. Thus, net effect of biotic interactions is often positive in stressed grasslands, where dominant species and litter can protect the subordinate species. Besides facilitation, niche partitioning can also support species coexistence leading to limiting similarity between subordinate species. Our aim was to provide a detailed analysis of fine-scale biotic interactions in stressed alkali grasslands. We supposed, that there are positive relationships between the main biomass fractions and species richness. We expected the expansion of trait ranges and the increase of trait dissimilarity with increasing biomass scores (total litter, green biomass of dominant species) and species richness. We studied the relationships between main biomass fractions, species richness, functional diversity and functional trait indices (ranges, weighted means and Rao indices). We used fine-scale biomass sampling in nine stands of dry alkali grasslands dominated by Festuca pseudovina. The detected relationships were always positive between the main biomass fractions (green biomass of dominant species, total litter and green biomass of subordinate species) and species richness. We found that the green biomass of dominant species and total litter increased ranges and dissimilarity of functional traits. Our results suggest that in dry alkali grasslands facilitation is crucial in shaping vegetation composition. The green biomass of dominant species and total litter increased the biomass production of subordinate species leading to overyielding. We found that mechanisms of facilitation and limiting similarity were jointly shaping the species coexistence in stressed grasslands, such as alkali grasslands.     

Dispersal traits as indicators of vegetation dynamics in long-term old-field succession

Succession is a key ecological process that supports our understanding of community assembly and biotic interactions. Dispersal potential and dispersal strategies, such as wind- or animal-dispersal, have been assumed to be highly relevant for the success of plant species during succession. However, research yielded varying results on changes in dispersal modes between successional stages. Here, we test the hypotheses that (a) vascular plant species that use a number of dispersal modes dominate in early stages of succession while species specialized on one/few dispersal modes increase in abundance towards later stages of succession; (b) species well adapted to wind-dispersal (anemochory) will peak in abundance in early successional stages and (c) species well adapted to adhesive dispersal (epizoochory) will increase with proceeding succession. We test these hypotheses in four sites within agriculturally dominated landscapes in Germany. Agricultural use in these sites was abandoned 20–28 years ago, leaving them to secondary succession. Sites have been monitored for plant biodiversity ever since. We analyze changes in plant species richness and abundance, number of dispersal modes and two ranking indices for wind- and adhesive dispersal by applying generalized linear mixed-effect models. We used both abundance-weighted and unweighted dispersal traits in order to gain a comprehensive picture of successional developments. Hypothesis (a) was supported by unweighted but not abundance-weighted data. Anemochory showed no consistent changes across sites. In contrast, epizoochory (especially when not weighted by abundance) turned out to be an indicator of the transition from early to mid-successional stages. It increased for the first 9–16 years of succession but declined afterwards. Species richness showed an opposing pattern, while species abundance increased asymptotically. We suggest that plant-animal interactions play a key role in mediating these processes: By importing seeds of highly competitive plant species, animals are likely to promote the increasing abundance of a few dominant, highly epizoochorous species. These species outcompete weak competitors and species richness decreases. However, animals should as well promote the subsequent increase of species richness by disturbing the sites and creating small open patches. These patches are colonized by weaker competitors that are not necessarily dispersed by animals. The changes in the presence of epizoochorous species indicate the importance of plant traits and related plant–animal interactions in the succession of plant communities.     

Dosimetric performance of continuous EPID imaging in stereotactic treatment conditions

PurposeThis study aims at investigating the dosimetric characteristics of a Varian aS1000 EPID, focusing on its continuous acquisition mode under the challenging conditions that can be met in stereotactic radiotherapy verification.MethodsAn aS1000 EPID installed on a Varian TrueBeamSTx was irradiated with 6 and 10 MV unflattened and flattened photon beams. In order to avoid detector saturation, the source-to-detector distance (SDD) was set to 150 or 180 cm depending on the dose rate. EPID image sets were acquired in continuous mode (CM) and also in the commonly used integrated mode (IM) for comparison, to evaluate dose linearity (including dose rate dependence), repeatability, reproducibility, stability, ghosting effect and field size dependence.ResultsCM response linearity was found to be within 0.8% of IM and independent of dose rate. Response repeatability was slightly better for IM and FF beams, being in all cases within 0.9%. Reproducibility was within 0.6% for both modes and all beam qualities. Response stability between continuous frames varied within 1% for dynamic and static irradiations and for all the beam qualities, showing its independence from these parameters. Ghosting effect was not significant, being comparable to signal variations between continuous frames (±1%). Field size dependence in both modes agreed within 1%.ConclusionsThe dosimetric response of the aS1000 EPID in CM with FFF beams and high dose rates is comparable to that in IM and for flattened beams provided that the appropriate SDD is used. aS1000 EPID in continuous acquisition mode is therefore suitable for stereotactic applications.     

Mouse embryos exposed to oxygen concentrations that mimic changes in the oviduct and uterus show improvement in blastocyst rate,blastocyst size,and accelerated cell division

After in vivo fertilisation, the preimplantation embryo goes through cleavage during migration along the oviduct in mammals or the fallopian tube in a woman and ends up inside the uterus. This study investigates the effect of a protocol aimed at closely reproducing that natural oxygen concentration in the oviduct (7 % O₂ from day 1 to day 3 and 2 % from day 3 to day 5), in contrast to the concentrations (5 % or 20 %) widely used in practice in ART using morphokinetic. Female mice (BI6/CBAca) were sacrificed, and zygotes were isolated 20 h after mating and randomly allocated to three parallel groups, which were grown under high atmospheric, low, or sequential oxygen concentrations. Zygotes were cultured in GTL medium (Vitrolife) and observed by the Primovision time-lapse system. Blastocyst rate at 120 h in the sequential group (91.3 %) was significantly increased over the high (76.3 %) and low (74.4 %) groups. Blastocyst size was also enlarged in the sequential group compared to the high and low groups. Moreover, cell division in the sequential group was significantly faster at almost every cleavage stage than it was in the other groups. Notably, the duration of the interims between stages also differed significantly between the groups. This study demonstrated that, in comparison to routinely used high or low oxygen conditions, oxygen concentrations mimicking changes in the oviduct and uterus significantly improve the blastocyst rate and size and accelerate cell division at several stages as well as the interims between cleavage events.     

毛竹SWEET基因家族的全基因组鉴定与分析

糖外排转运蛋白(Sugars will eventually be exported transporters, SWEETs)在植物生理活动和发育过程中起重要作用。为探究SWEET基因家族在毛竹(Phyllostachys edulis)的生长发育过程中起的作用,基于毛竹基因组数据,通过生物信息学方法对SWEET基因家族成员进行鉴定,并对其编码的蛋白质理化性质、系统进化及共线性关系、基因结构、启动子元件及表达模式、蛋白互作网路分析、GO注释等进行细致分析。研究结果表明:该家族基因结构、基序和结构域相对保守,所有成员均含有MtN3_slv结构域。上游启动子序列中含有多个同非生物胁迫以及生长发育相关元件,结合转录组表达量分析显示,多个家族成员在毛竹不同组织器官均有表达。共线性分析揭示毛竹SWEET家族在演化过程中存在全基因组多倍化事件。蛋白互作网路分析挖掘出2个重要核心家族成员,GO注释分析进一步证实毛竹SWEET主要负责体内糖类物质的转运。以上结果为毛竹SWEET基因功能鉴定提供了重要参考,对于毛竹快速生长分子机制研究打下了基础。     

Gadolinium-containing carbon nanomaterials for magnetic resonance imaging: Trends and challenges

Gadolinium-containing carbon nanomaterials are a new class of contrast agent for magnetic resonance imaging. They are characterized by a superior proton relaxivity to any current commercial gadolinium contrast agent and offer the possibility to design multifunctional contrasts. Intense efforts have been made to develop these nanomaterials because of their potential for better results than the available gadolinium contrast agents. The aim of the present work is to provide a review of the advances in research on gadolinium-containing carbon nanomaterials and their advantages over conventional gadolinium contrast agents. Due to their enhanced proton relaxivity, they can provide a reliable imaging contrast for cells, tissues or organs with much smaller doses than currently used in clinical practice, thus leading to reduced toxicity (as shown by cytotoxicity and biodistribution studies). Their active targeting capability allows for improved MRI of molecular or cellular targets, overcoming the limited labelling capability of available contrast agents (restricted to physiological irregularities during pathological conditions). Their potential of multifunctionality encompasses multimodal imaging and the combination of imaging and therapy.     

Microfluidic‐Assisted Blade Coating of Compositional Libraries for Combinatorial Applications: The Case of Organic Photovoltaics

Microfluidic technologies are highly adept at generating controllable compositional gradients in fluids, a feature that has accelerated the understanding of the importance of chemical gradients in biological processes. That said, the development of versatile methods to generate controllable compositional gradients in the solid‐state has been far more elusive. The ability to produce such gradients would provide access to extensive compositional libraries, thus enabling the high‐throughput exploration of the parametric landscape of functional solids and devices in a resource‐, time‐, and cost‐efficient manner. Herein, the synergic integration of microfluidic technologies is reported with blade coating to enable the controlled formation of compositional lateral gradients in solution. Subsequently, the transformation of liquid‐based compositional gradients into solid‐state thin films using this method is demonstrated. To demonstrate efficacy of the approach, microfluidic‐assisted blade coating is used to optimize blending ratios in organic solar cells. Importantly, this novel technology can be easily extended to other solution processable systems that require the formation of solid‐state compositional lateral gradients.     

Efficient Exploration of the Composition Space in Ternary Organic Solar Cells by Combining High‐Throughput Material Libraries and Hyperspectral Imaging

Organic solar cells based on ternary active layers can lead to higher power conversion efficiencies than corresponding binaries, and improved stability. The parameter space for optimization of multicomponent systems is considerably more complex than that of binaries, due to both, a larger number of parameters (e.g., two relative compositions rather than one) and intricate morphology–property correlations. Most experimental reports to date reasonably limit themselves to a relatively narrow subset of compositions (e.g., the 1:1 donor/s:acceptor/s trajectory). This work advances a methodology that allows exploration of a large fraction of the ternary phase space employing only a few (<10) samples. Each sample is produced by a designed sequential deposition of the constituent inks, and results in compositions gradients with ≈5000 points/sample that cover about 15%–25% of the phase space. These effective ternary libraries are then colocally imaged by a combination of photovoltaic techniques (laser and white light photocurrent maps) and spectroscopic techniques (Raman, photoluminescence, absorption). The generality of the methodology is demonstrated by investigating three ternary systems, namely PBDB‐T:ITIC:PC₇₀BM, PTB7‐Th:ITIC:PC₇₀BM, and P3HT:O‐IDFBR:O‐IDTBR. Complex performance‐structure landscapes through the ternary diagram as well as the emergence of several performance maxima are discovered.     

用于可视化光治疗的Mn3O4@CuS纳米粒的制备研究

摘要 目的：本研究旨在制备用于肿瘤可视化光治疗的多功能Mn₃O₄@CuS核壳型纳米粒,在磁共振成像的引导下,使用近红外光定点辐照,实现局部光热消融治疗。方法：（1）采用高温热解法制备油胺稳定的Mn₃O₄纳米粒,在其表面构建CuS壳层,并进行聚乙二醇修饰,得到分散于水相中的Mn₃O₄@CuS核壳型纳米粒。（2）通过透射电镜、紫外可见近红外吸收光谱等方法对该纳米粒进行理化性质表征,并研究其体外磁共振成像、光热升温等性能。结果：制备的水相分散的Mn₃O₄@CuS纳米粒,粒径均一且分散性较好,形态为近圆形,粒径为9.30±2.29 nm;紫外可见近红外吸收光谱图表明Mn₃O₄@CuS纳米粒在近红外区有较强吸收,最大吸收峰位于1100~1200 nm范围;磁共振成像分析结果可计算出Mn₃O₄@CuS纳米粒的纵向弛豫率r1为1.662 mM^-1s^-1,表明其具有较好的磁共振增强造影效果;光热升温曲线显示Mn₃O₄@CuS纳米粒可在785 nm近红外激光下升温至73.5 ℃,具备较好的光热治疗潜力。结论：本文成功制备出水相分散的Mn₃O₄@CuS核壳型纳米粒,具有良好的磁共振造影成像性能和光热升温效应,有望应用于磁共振成像引导下的肿瘤可视化光治疗。