The P2Y2 Receptor C-Terminal Tail Modulates but Is Dispensable for β-Arrestin Recruitment

The P2Y₂ receptor (P2Y₂R) is a G protein-coupled receptor that is activated by extracellular ATP and UTP, to a similar extent. This allows it to play roles in the cell’s response to the (increased) release of these nucleotides, e.g., in response to stress situations, including mechanical stress and oxygen deprivation. However, despite its involvement in important (patho)physiological processes, the intracellular signaling induced by the P2Y₂R remains incompletely described. Therefore, this study implemented a NanoBiT^® functional complementation assay to shed more light on the recruitment of β-arrestins (βarr1 and βarr2) upon receptor activation. More specifically, upon determination of the optimal configuration in this assay system, the effect of different (receptor) residues/regions on βarr recruitment to the receptor in response to ATP or UTP was estimated. To this end, the linker was shortened, the C-terminal tail was truncated, and phosphorylatable residues in the third intracellular loop of the receptor were mutated, in either singly or multiply adapted constructs. The results showed that none of the introduced adaptations entirely abolished the recruitment of either βarr, although EC₅₀ values differed and time-luminescence profiles appeared to be qualitatively altered. The results hint at the C-terminal tail modulating the interaction with βarr, while not being indispensable.     

Enhancing Post-Stroke Rehabilitation and Preventing Exo-Focal Dopaminergic Degeneration in Rats—A Role for Substance P

Dopaminergic signaling is a prerequisite for motor learning. Delayed degeneration of dopaminergic neurons after stroke is linked to motor learning deficits impairing motor rehabilitation. This study investigates safety and efficacy of substance P (SP) treatment on post-stroke rehabilitation, as this neuropeptide combines neuroprotective and plasticity-promoting properties. Male Sprague Dawley rats received a photothrombotic stroke within the primary motor cortex (M1) after which a previously acquired skilled reaching task was rehabilitated. Rats were treated with intraperitoneal saline (control group, n = 7) or SP-injections (250 µg/kg) 30 min before (SP-pre; n = 7) or 16 h (SP-post; n = 6) after rehabilitation training. Dopaminergic neurodegeneration, microglial activation and substance P-immunoreactivity (IR) were analyzed immunohistochemically. Systemic SP significantly facilitated motor rehabilitation. This effect was more pronounced in SP-pre compared to SP-post animals. SP prevented dopaminergic cell loss after stroke, particularly in the SP-pre condition. Despite its proinflammatory propensity, SP administration did not increase stroke volumes, post-stroke deficits or activation of microglia in the midbrain. Finally, SP administration prevented ipsilesional hypertrophy of striatal SPergic innervation, particularly in the SP-post condition. Mechanistically, SP-pre likely involved plasticity-promoting effects in the early phase of rehabilitation, whereas preservation of dopaminergic signaling may have ameliorated rehabilitative success in both SP groups during later stages of training. Our results demonstrate the facilitating effect of SP treatment on motor rehabilitation after stroke, especially if administered prior to training. SP furthermore prevented delayed dopaminergic degeneration and preserved physiological endogenous SPergic innervation.     

Identification and In Silico Characterization of a Novel COLGALT2 Gene Variant in a Child with Mucosal Rectal Prolapse

Rectal prolapse is influenced by many factors including connective tissue dysfunction. Currently, there is no data about genetic contribution in the etiology of this disorder. In this study, we performed trio whole-exome sequencing in an 11-year-old girl with mucosal rectal prolapse and her parents and sibling. Genetic testing revealed a novel heterozygous missense variant c.1406G>T; p.G469V in exon 11 of the COLGALT2 gene encoding the GLT25 D2 enzyme. Sanger sequencing confirmed this variant only in the patient while the mother, father and sister showed a wild-type sequence. The pathogenicity of the novel variant was predicted using 10 different in silico tools that classified it as pathogenic. Further, in silico prediction, according to Phyre2, Project HOPE, I-Mutant3.0 and MutPred2 showed that the missense variant can decrease protein stability and cause alterations in the physical properties of amino acids resulting in structural and functional changes of the GLT25D2 protein. In conclusion, the present study identifies a previously unknown missense mutation in the COLGALT2 gene that encodes the enzyme involved in collagen glycosylation. The clinical features observed in the patient and the results of in silico analysis suggest that the new genetic variant can be pathogenic.     

利用响应面分析法优化高山被孢霉M E - 1 生产花生四烯酸培养基成分研究

在利用高山被孢霉ME-1 生产花生四烯酸(ARA)过程中,采用响应面分析法,对摇瓶中的培养基成分进行优化。建立了两个标准的多项式模型：在长达6.5d 发酵过程中,当葡萄糖、酵母膏、KH2PO4 和NaNO3 浓度分别为90.16、12.50、3.80 和3.54g/L 时,生物量将到最大,约36.86g/L;当葡萄糖、酵母膏、KH2PO4 和NaNO3浓度分别为103.16、11.66、3.80 和3.43g/L 时,AA 产量将到最大,约9.65g/L。预测值通过实验得到了充分的证实,预测值和实验结果相关性很好。发酵罐实验结果表明,在高山被孢霉ME-1 大规模发酵生产过程中,对培养基进行优化,将同时引起生物量(发酵5d,约34.21 ± 1.01g/L)和AA 产量(发酵6d,约9.86 ± 0.45g/L)的增加。     

M5纤维的研究及应用

介绍了M5纤维的结构、物理性能、抗冲击性、阻燃性及其与其他几种高强高模纤维的性能对比,概述了M5纤维在复合材料、个体防护用品、防火产品、绳索和先进织物上的应用。     

Effects of ready to drink teas on AhR- and PXR-mediated expression of cytochromes P450 CYP1A1 and CYP3A4 in human cancer cell lines and primary human hepatocytes

A variety of xenobiotics are taken in the diet and they can interfere with regulatory pathways of drug metabolizing enzymes in humans. This can result in food-drug interactions, which is undesirable clinical situation where drug pharmacokinetics are influenced by dietary compounds. Xenobiotics-mediated food-drug interactions include the induction of drug metabolizing cytochromes P450. The expression of the most important inducible cytochromes CYP1A and CYP3A4 are regulated by xenoreceptors PXR and AhR.We examined extracts from 17 different flavoured ready to drink teas (RDTs) for their capabilities to activate PXR and AhR receptors and to induce CYP3A4 and CYP1A genes. Primary cultures of human hepatocytes and cancer cell lines HepG2 and LS174T were used as in vitro models. Gene reporter assays, RT-PCR and Western blots were performed.We identified three RDTs that induced CYP3A4 mRNA and protein, implying a potential for food-drug interactions. Several RDTs slightly elevated CYP1A1 expression or activated AhR.     

对激光光束质量一些问题的认识

从对激光光束质量研究进展开始,讨论了在傍轴和非傍轴范畴内激光光束质量的一些问题,主要是非傍轴光束的光束质量评价。光束传输的非傍轴矢量二阶矩理论可用于计算多种非傍轴光束的M2因子,但对M2因子的物理解释却有困难。例如,在非傍轴范畴,M2因子依赖于束腰宽度与波长之比,并可小于1。但远场发散角之值可趋于90°,这将导致很大的M2因子值。另一方面,若将公式中的光强用时间平均坡印廷矢量z分量代替,桶中功率(PIB),或等价地,?茁 参数可推广用于评价非傍轴光束的远场光束质量。PIB表征了光束功率的可聚焦能力,是一种简明、直观评价不同激光光束的方法。然而,在?茁参数的公式中的束宽是按功率含量百分比定义的,因此双曲线传输规律不再严格成立。     

“配套性”民用衣柜的“标准化”延伸柜探究

本文通过“32mm系统”家具设计方法在衣柜设计中的运用,提供了—种具有普适性的模块化设计准则。意在引导—种能将家具的结构与功能进行有机结合,同时,兼顾材料与外观的标准化、通用化、系列化设计理念,从而达到与国际行业规范相接轨的目的。     

抗黄曲霉毒素M1单克隆抗体细胞株的筛选

研究筛选到抗黄曲霉毒素M1的单克隆抗体细胞株,用AFM1-oxim e-BSA偶联物免疫Balb/C小鼠后,取小鼠脾细胞与Sp2/0骨髓瘤细胞融合,经过3次克隆和亚克隆得到两株稳定分泌抗黄曲霉毒素M1抗体的杂交瘤细胞株,经过检测两株均为IgG1亚类,并且经过鉴定其各项性质均符合标准。     

苔干水溶性粗多糖提取工艺及理化性质的研究

本文采用热水浸提、醇沉法提取苔干多糖,对苔干多糖提取工艺和一般理化性质进行了研究。选择浸提固液比、温度和时间作为单因素进行梯度实验,确定其条件范围,再通过正交实验L9(34)进一步确定苔干水溶性粗多糖最佳提取工艺条件。结果表明最佳提取工艺为:固液比1:40、浸提温度90℃、浸提时间3h、反复浸提2次。理化性质结果表明提取的苔干多糖中含有多肽或氨基酸,为蛋白质复合多糖。