Integrity and Regeneration of Mechanotransduction Machinery Regulate Aminoglycoside Entry and Sensory Cell Death

Sound perception requires functional hair cell mechanotransduction (MET) machinery, including the MET channels and tip-link proteins. Prior work showed that uptake of ototoxic aminoglycosides (AG) into hair cells requires functional MET channels. In this study, we examined whether tip-link proteins, including Cadherin 23 (Cdh23), regulate AG entry into hair cells. Using time-lapse microscopy on cochlear explants, we found rapid uptake of gentamicin-conjugated Texas Red (GTTR) into hair cells from three-day-old Cdh23^+/+ and Cdh23^v2J/+ mice, but failed to detect GTTR uptake in Cdh23^v2J/v2J hair cells. Pre-treatment of wildtype cochleae with the calcium chelator 1,2-bis(o-aminophenoxy) ethane-N,N,N'',N''-tetraacetic acid (BAPTA) to disrupt tip-links also effectively reduced GTTR uptake into hair cells. Both Cdh23^v2J/v2J and BAPTA-treated hair cells were protected from degeneration caused by gentamicin. Six hours after BAPTA treatment, GTTR uptake remained reduced in comparison to controls; by 24 hours, drug uptake was comparable between untreated and BAPTA-treated hair cells, which again became susceptible to cell death induced by gentamicin. Together, these results provide genetic and pharmacologic evidence that tip-links are required for AG uptake and toxicity in hair cells. Because tip-links can spontaneously regenerate, their temporary breakage offers a limited time window when hair cells are protected from AG toxicity.     

丽江老君山海拔上限长苞冷杉(Abies georgei)和云南铁杉(Tsuga dumosa)径向生长对气候变化的响应

基于树木年轮学方法,利用丽江老君山海拔上限长苞冷杉（Abies georgei）和云南铁杉（Tsuga dumosa）树轮宽度资料,构建差值年表,运用响应函数和滑动响应分析研究树木径向生长与气温和降水的相关关系及其稳定性,进而阐明影响该区域2个针叶树种径向生长的主要气候要素。结果表明：2个树种对降水累积效应的响应较为一致,对逐月气候因子的响应存在差异,相关关系较为稳定,具体表现为（1）上年11月平均温升高和当年生长季盛期（7-8月）降水增加有利于老君山海拔上限长苞冷杉生长;（2）云南铁杉径向生长与当年3月、树木休眠期（1-3月）、生长季盛期（7-8月）的降水表现为显著正相关关系,与上年7月与当年5月的气温及当年生长季末期（9-10月）降水呈显著负相关;（3）上述相关关系的稳定性较强,在全部或大部分分析时段（1951-2017）内达到显著相关,云南铁杉的稳定性更强。研究结果可为气候变化背景下滇西北高原树木生长的管理及森林生态系统的保护提供理论依据。     

Candida albicans Elicits Pro-Inflammatory Differential Gene Expression in Intestinal Peyer’s Patches

The details of how gut-associated lymphoid tissues such as Peyer’s patches (PPs) in the small intestine play a role in immune surveillance, microbial differentiation and the mucosal barrier protection in response to fungal organisms such as Candida albicans are still unclear. We particularly focus on PPs as they are the immune sensors and inductive sites of the gut that influence inflammation and tolerance. We have previously demonstrated that CD11c⁺ phagocytes that include dendritic cells and macrophages are located in the sub-epithelial dome within PPs sample C. albicans. To gain insight on how specific cells within PPs sense and respond to the sampling of fungi, we gavaged naïve mice with C. albicans strains ATCC 18804 and SC5314 as well as Saccharomyces cerevisiae. We measured the differential gene expression of sorted CD45⁺ B220⁺ B-cells, CD3⁺ T-cells and CD11c⁺ DCs within the first 24 h post-gavage using nanostring nCounter^® technology. The results reveal that at 24 h, PP phagocytes were the cell type that displayed differential gene expression. These phagocytes were able to sample C. albicans and discriminate between strains. In particular, strain ATCC 18804 upregulated fungal-specific pro-inflammatory genes pertaining to innate and adaptive immune responses. Interestingly, PP CD11c⁺ phagocytes also differentially expressed genes in response to C. albicans that were important in the protection of the mucosal barrier. These results highlight that the mucosal barrier not only responds to C. albicans, but also aids in the protection of the host.

     

Probing intrinsic properties of a robust morphogen gradient in Drosophila

A remarkable feature of development is its reproducibility, the ability to correct embryo-to-embryo variations and instruct precise patterning. In Drosophila, embryonic patterning along the anterior-posterior axis is controlled by the morphogen gradient Bicoid (Bcd). In this article, we describe quantitative studies of the native Bcd gradient and its target Hunchback (Hb). We show that the native Bcd gradient is highly reproducible and is itself scaled with embryo length. While a precise Bcd gradient is necessary for precise Hb expression, it still has positional errors greater than Hb expression. We describe analyses further probing mechanisms for Bcd gradient scaling and correction of its residual positional errors. Our results suggest a simple model of a robust Bcd gradient sufficient to achieve scaled and precise activation of its targets. The robustness of this gradient is conferred by its intrinsic properties of "self-correcting" the inevitable input variations to achieve a precise and reproducible output.     

Virtual screening and biophysical studies lead to HSP90 inhibitors

Heat shock protein 90 (HSP90) is a molecular chaperone that plays important functional roles in cells. The chaperone activity of HSP90 is regulated by the hydrolysis of ATP at the protein’s N-terminal domain. HSP90, in particular the N-terminal domain, is a current inhibition target for therapeutic treatments of cancers. This paper describes an application of virtual screening, thermal shift assaying and protein NMR spectroscopy leading to the discovery of HSP90 inhibitors that contain the resorcinol structure. The resorcinol scaffold can be found in a class of HSP90 inhibitors that are currently undergoing clinical trials. The proved success of the resorcinol moiety in HSP90 inhibitors validates this combined virtual screen and biophysical technique approach, which may be applied for future inhibitor discovery work for HSP90 as well as other targets.     

Size‐dependent dual emission of Cu,Mn:ZnSe QDs: Controlling both emission wavelength and intensity

Cu,Mn:ZnSe quantum dots (QDs) of tunable size, controllable photoluminescence (PL) intensity ratio and PL range were prepared. A study of the experimental conditions confirmed that the size of Cu,Mn:ZnSe QDs is affected by the pH of the solution, the speed at which the Zn solution is injected and the reaction temperature. In general, high pH, low injection speed and high reaction temperature are optimal for preparing large QDs. Based on this knowledge, different sizes of Cu,Mn:ZnSe QDs were synthesized. Moreover, white emission Cu,Mn:ZnSe QDs were designed by controlling the experimental conditions and the feeding mole ratio of Mn:Cu.     

Composition and structure of nucleolar skeleton (nucleolar matrix)-Actin and fibrillarin are two main protein components of nucleolar skeleton

Purified nucleoli of HeLa cells were treated sequentially with nonionic detergent, nucleic acid enzyme, low salt and high salt. The residual nucleolar structure termed nucleolar skeleton (nucleolar matrix) was shown as a fine network under electron microscope with DGD embedding-unembedding technique. Such structures of BHK-21 cell and mouse liver cell are similar to that of HeLa cell. The protein composition of the nucleolar skeleton of HeLa cells was analyzed. The protein composition of such nucleolar residual shows obvious difference from the compositions of nuclear matrix and chromosome scaffold. The major protein composition of the nucleolar skeleton of HeLa cells contains 6–7 polypeptides. Their molecular weights are about 48, 43, 36 and 33 ku. Further studies show that actin and fibrillarin are two major protein components of nucleolar skeleton of HeLa cells.     

基于多源遥感影像的盐碱地治理效果

将Landsat-8 OLI影像与Sentinel-2号影像结合,利用光合有效辐射比率（FPAR）,构建适用于MODIS植被总初级生产力（GPP）影像空间分辨率提高的逐层逐像元修正因子,实现内蒙古巴彦淖尔市赵贵圪旦组盐碱地农田改良前后的植被长势监测,同时结合不同深度土壤的含盐量数据,采用转移矩阵方法与热点分析方法分别分析了全局土壤含盐量变化与植被长势变化的局部差异。结果表明：（1）12-20号地块内土壤含盐量改良效果最明显,且研究区改良效果随土壤深度增加而逐渐降低。土壤深度0-20cm,盐土向中度盐土转入面积最大;土壤深度20-40cm,强度盐土向中度盐土转入面积最大;土壤深度40-60cm,强度盐土与盐土的转入面积大于转出面积,其他土壤的转入面积小于转出面积。（2）研究区内,GPP提升区域比降低区域面积约多60.36hm²,占研究区面积的42.88%;同时,对改良前植被GPP低值区域提高效果更明显。（3） GPP提升区域中热点区域集中分布在12-17、25-27、34号地块内,该区域改良效果显著;GPP降低区域中冷点区域集中分布在6、12-14、22-23、27-28、30-31号地块内,该区域改良效果差。     

Cationicity-Enhanced Analogues of the Antimicrobial Peptides,AcrAP1 and AcrAP2, from the Venom of the Scorpion,Androctonus crassicauda,Display Potent Growth Modulation Effects on Human Cancer Cell Lines

The non disulphide-bridged peptides (NDBPs) of scorpion venoms are attracting increased interest due to their structural heterogeneity and broad spectrum of biological activities. Here, two novel peptides, named AcrAP1 and AcrAP2, have been identified in the lyophilised venom of the Arabian scorpion, Androctonus crassicauda, through “shotgun” molecular cloning of their biosynthetic precursor-encoding cDNAs. The respective mature peptides, predicted from these cloned cDNAs, were subsequently isolated from the same venom sample using reverse phase HPLC and their identities were confirmed by use of mass spectrometric techniques. Both were found to belong to a family of highly-conserved scorpion venom antimicrobial peptides - a finding confirmed through the biological investigation of synthetic replicates. Analogues of both peptides designed for enhanced cationicity, displayed enhanced potency and spectra of antimicrobial activity but, unlike the native peptides, these also displayed potent growth modulation effects on a range of human cancer cell lines. Thus natural peptide templates from venom peptidomes can provide the basis for rational analogue design to improve both biological potency and spectrum of action. The diversity of such templates from such natural sources undoubtedly provides the pharmaceutical industry with unique lead compounds for drug discovery.