Isolation of Decidual Macrophages and Hofbauer Cells from Term Placenta—Comparison of the Expression of CD163 and CD80

(1) Background: Placental immune cells are playing a very important role in a successful placentation and the prevention of pregnancy complications. Macrophages dominate in number and relevance in the maternal and the fetal part of the placenta. The evidence on the polarization state of fetal and maternal macrophages involved in both, healthy and pregnancy-associated diseases, is limited. There is no representative isolation method for the direct comparison of maternal and fetal macrophages so far. (2) Material and Methods: For the isolation of decidual macrophages and Hofbauer cells from term placenta, fresh tissue was mechanically dissected and digested with trypsin and collagenase A. Afterwards cell enrichment was increased by a Percoll gradient. CD68 is represented as pan-macrophage marker, the surface markers CD80 and CD163 were further investigated. (3) Results: The established method revealed a high cell yield and purity of the isolated macrophages and enabled the comparison between decidual macrophages and Hofbauer cells. No significant difference was observed in the percentage of single CD163⁺ cells in the distinct macrophage populations, by using FACS and immunofluorescence staining. A slight increase of CD80⁺ cells could be found in the decidual macrophages. Considering the percentage of CD80⁺CD163⁻ and CD80⁻CD163⁺ cells we could not find differences. Interestingly we found an increased number of double positive cells (CD80⁺CD163⁺) in the decidual macrophage population in comparison to Hofbauer cells. (4) Conclusion: In this study we demonstrate that our established isolation method enables the investigation of decidual macrophages and Hofbauer cells in the placenta. It represents a promising method for direct cell comparison, enzyme independently, and unaffected by magnetic beads, to understand the functional subsets of placental macrophages and to identify therapeutic targets of pregnancy associated diseases.     

Hyperinsulinemic Hypoglycemia Associated with a CaV1.2 Variant with Mixed Gain- and Loss-of-Function Effects

The voltage-dependent L-type calcium channel isoform Ca_V1.2 is critically involved in many physiological processes, e.g., in cardiac action potential formation, electromechanical coupling and regulation of insulin secretion by beta cells. Gain-of-function mutations in the calcium voltage-gated channel subunit alpha 1 C (CACNA1C) gene, encoding the Ca_V1.2 α₁-subunit, cause Timothy syndrome (TS), a multisystemic disorder that includes autism spectrum disorders and long QT (LQT) syndrome. Strikingly, TS patients frequently suffer from hypoglycemia of yet unproven origin. Using next-generation sequencing, we identified a novel heterozygous CACNA1C mutation in a patient with congenital hyperinsulinism (CHI) and associated hypoglycemic episodes. We characterized the electrophysiological phenotype of the mutated channel using voltage-clamp recordings and in silico action potential modeling experiments. The identified Ca_V1.2^L566P mutation causes a mixed electrophysiological phenotype of gain- and loss-of-function effects. In silico action potential modeling supports that this mixed electrophysiological phenotype leads to a tissue-specific impact on beta cells compared to cardiomyocytes. Thus, CACNA1C variants may be associated with non-syndromic hyperinsulinemic hypoglycemia without long-QT syndrome, explained by very specific electrophysiological properties of the mutated channel. We discuss different biochemical characteristics and clinical impacts of hypoglycemia in the context of CACNA1C variants and show that these may be associated with significant morbidity for Timothy Syndrome patients. Our findings underline that the potential of hypoglycemia warrants careful attention in patients with CACNA1C variants, and such variants should be included in the differential diagnosis of non-syndromic congenital hyperinsulinism.     

Using Drosophila melanogaster to Analyse the Human Paralogs of the ESCRT-III Core Component Shrub/CHMP4/Snf7 and Its Interactions with Members of the LGD/CC2D1 Family

The evolutionary conserved ESCRT-III complex is a device for membrane remodelling in various cellular processes, such as the formation of intraluminal vesicles (ILVs), cytokinesis, and membrane repair. The common theme of all these processes is the abscission of membrane away from the cytosol. At its heart in Drosophila is Shrub, CHMP4 in humans, which dynamically polymerises into filaments through electrostatic interactions among the protomers. For the full activity, Shrub/CHMP4 requires physical interaction with members of the Lgd protein family. This interaction is mediated by the odd-numbered DM14 domains of Lgd, which bind to the negative interaction surface of Shrub. While only one Lgd and one Shrub exist in the genome of Drosophila, mammals have two Lgd orthologs, LGD1/CC2D1B and LGD2/CC2D1A, as well as three CHMP4s in their genomes, CHMP4A, CHMP4B, and CHMP4C. The rationale for the diversification of the ESCRT components is not understood. We here use Drosophila as a model system to analyse the activity of the human orthologs of Shrub and Lgd at an organismal level. This enabled us to use the plethora of available techniques available for Drosophila. We present evidence that CHMP4B is the true ortholog of Shrub, while CHMP4A and CHMP4C have diverging activities. Nevertheless, CHMP4A and CHMP4C can enhance the activity of CHMP4B, raising the possibility that they can form heteropolymers in vivo. Our structure-function analysis of the LGD1 and LGD2 indicates that the C2 domain of the LGD proteins has a specific function beyond protein stability and subcellular localisation. Moreover, our data specify that CHMP4B interacts more efficiently with LGD1 than with LGD2.     

采用分布式孔径的红外敏感器系统（上）

第三代焦平面列阵与能力越来越强的信号处理算法的结合,正在使电光技术不断走向成熟,同时也给军用和非军用电光敏感器系统的设计指出了新的方向,目前,美国国防部正在利用这些进展研制一种可安装在各种战术平台上的分布式孔径红外敏感器系统（DAIRS）,这种DAIRS是利用多个相同的固定敏感器来获得其功能度的,而在以前,这种功能度是通过对每一种功能都使用专用的敏感器来获得的,在战术飞机上,DAIRS的作用是利用一个包含六个按战略要求的敏感器列阵,向飞行人员提供4π立体弧度的敏感器覆盖范围,即全球面情况察觉（SA）,这种察觉提供：导弹威胁告警,红外搜索与跟踪,战斗损害估计,目标导向辅助以及领航。DAIRS还可以为水面舰艇,装甲登陆车以及无人驾驶空战飞行器提供扩展的全球面情况察觉。敏感器的重量,体积以及电功率需求,一般不到现有机载红外敏感器系统的25％,而且系统实际运行成本可以大为降低,如果与自动提示技术相结合,DAIRS就有可能在用于空中防撞的飞机防撞告警的系统技术发展方面起重要作用。DAIRS目前得到了海军研究局的部分资助,这将可以生产两台原型敏感器和实验室测试所需的支持软件,多功能红外分布式孔径系统（MIDAS）是作为一项海军先进技术演示（6．3计划）而获得资金的,根据此项计划,DAIRS将于2000年用四个敏感器进行飞行试验,MIDAS将配备一种装在头盔上的用于情况察觉成像的显示器。     

A field-portable thermal infrared grating spectrometer (THIRSPEC)

A thermal infrared grating spectrometer was developed for field studies in the Earth sciences. The design is based on a reflection grating and a 60-element HgCdTe detector array. The useful spectral range of the instrument covers 7.9-11.3 μm with a Nyquist limited resolution of 0.16 μm. The instrument averages over a 12° field of view and compares the exitance of the target to that of an internal black body at ambient temperature. The noise equivalent temperature is approximately 0.06°K over the useful spectral range. Background radiance reflected from the surface of the target can seriously impede the determination of emissivity. This effect is removed from the spectra of geological samples by the use of reference samples     

External tools for collaborative medication scheduling

Medication adherence—taking medication as prescribed—is critical for successful self-care, especially among older adults. Adherence depends on developing and implementing plans for taking medications. Age-related cognitive declines that affect adherence may be mitigated using external tools that support patient-provider collaboration needed to develop these adherence plans. We tested the effectiveness of structured collaborative medication tools to support better medication planning and adherence practices. Evidence for benefits of collaborative tools has been mixed in terms of their usefulness for medication-scheduling tasks, perhaps because the tools have not been explicitly designed to support patient-provider collaboration. A total of 144 community-dwelling older adults participated in pairs and performed the role of a patient or provider in a simulated patient-provider medication-scheduling task. Each pair worked with a structured (MedTable^? and e-MedTable) or unstructured (Medcard) scheduling tool and completed four problems (2 simple and 2 complex). Performance was measured using the following: problem-solving (medication schedule) accuracy, problem-solving time, solution (schedule) optimality, tool usability, collaborative effectiveness, and subjective workload involved in creating the medication schedules. Participants using structured tools produced more accurate and optimal schedules. They also rated subjective workload as lower and thought that the structured tools were easier to use, reduced subjective workload associated with creating the schedules. There was also suggestive evidence that participants using the structured tools rated more highly the quality of their collaboration. Structured medication-scheduling tools have the potential to improve medication adherence among older adults because they support collaborative planning and reduce the cognitive load involved in creating these adherence plans.