The Mouse Round-window Approach for Ototoxic Agent Delivery: A Rapid and Reliable Technique for Inducing Cochlear Cell Degeneration

Investigators have utilized a wide array of animal models and investigative techniques to study the mammalian auditory system. Much of the basic research involving the cochlea and its associated neural pathways entails exposure of model cochleae to a variety of ototoxic agents. This allows investigators to study the effects of targeted damage to cochlear structures, and in some cases, the self-repair or regeneration of those structures. Various techniques exist for delivery of ototoxic agents to the cochlea. When selecting a particular technique, investigators must consider a number of factors, including the induction of inadvertent systemic toxicity, the amount of cochlear damage produced by the surgical procedure itself, the type of lesion desired, animal survivability, and reproducibility/reliability of results. Currently established techniques include parenteral injection, intra-peritoneal injection, trans-tympanic injection, endolymphatic sac injection, and cochleostomy with perilymphatic perfusion. Each of these methods has been successfully utilized and is well described in the literature; yet, each has various shortcomings. Here, we present a technique for topical application of ototoxic agents directly to the round window niche. This technique is non-invasive to inner ear structures, produces rapid onset of reliably targeted lesions, avoids systemic toxicity, and allows for an intra-animal control (the contra-lateral ear). Results stemming from this approach have helped deeper understanding of auditory pathophysiology, cochlear cell degeneration, and regenerative capacity in response to an acute injury. Future investigations may use this method to conduct interventional studies involving gene therapy and stem cell transplantation to combat hearing loss.     

不同去趋势方法对基于Dendrometer数据的茎干水分动态分析的影响——以白扦为例

树木茎干半径变化记录仪(Dendrometer)监测的高精度数据不仅包括木质部的年内径向生长过程, 还包含由茎干水分的消耗和补充引起的可逆变化。然而, 不同的年内生长去趋势方法获得的茎干水分波动之间的差异性仍缺乏对比研究。基于芦芽山北坡针叶林下限白扦(Picea meyeri) 2015年生长季的茎干半径变化和环境因子的实时监测数据, 使用Gompertz生长模型(GPZ)、线性生长模型(LG)、零生长模型(ZG)、日值法(D)和茎干循环法(SC)模拟并去除茎干年内的生长趋势, 然后提取5种不同类型树木水分缺乏引起的茎干收缩(TWD_GPZ、TWD_LG、TWD_ZG、TWD_D和TWD_SC)以表征茎干水分亏缺, 并进一步对比分析了不同茎干水分亏缺序列对环境中水分状况的响应特征。研究发现: (1)不同去趋势方法计算的茎干水分亏缺的趋势和幅度有所差异, 可聚类为3组: TWD_LG和TWD_ZG、TWD_GPZ以及TWD_D和TWD_SC。同组或聚类距离接近的序列在生长季内每个月份都展现出显著的相关性。然而, TWD_LG、TWD_ZG和TWD_GPZ与TWD_D和TWD_SC在8月份相关性较弱。(2) TWD_D和TWD_SC与空气饱和水汽压差(VPD)的正相关关系比TWD_GPZ、TWD_LG和TWD_ZG更加稳定, 且具有更大的相关系数。5种茎干水分亏缺序列和土壤含水量(SWC)的关系在生长季内变化很大。(3)不同去趋势方法的茎干水分亏缺都随着水分胁迫程度(VPD/SWC)升高而显著增长。当胁迫程度较低时, TWD_SC对VPD/SWC的变化最为敏感(R² = 0.39, p < 0.001), 但是与TWD_ZG差别不大(R ² = 0.37, p < 0.001); 当胁迫程度较高时, TWD_ZG对VPD/SWC的敏感性最高(R ² = 0.59, p < 0.001)。综合对比来看, 零生长模型是比较适合研究区白扦生长季内茎干水分波动的去趋势方法, 其可为干旱胁迫条件下预测研究区树木的茎干水分动态及特征提供科学 依据。     

Comparison of ultrafiltration units for proteomic and N-glycoproteomic analysis by the filter-aided sample preparation method

The precision with which the dissociation constant, K_D, can be obtained from isothermal titration calorimetry depends on, among other factors, the concentrations of the interacting species. The so-called c value—the ratio of analyte concentration to K_D—should fall in the range of 1 to 1000 for reliable K_D determination. On the basis of simulated, noise-free data, Biswas and Tsodikov [5] recently suggested an optimal c value of 5 to 20. By contrast, we find an optimum at c > 40 on determining the K_D confidence intervals through simulations containing noise levels typical of state-of-the-art microcalorimeters.     

Profiling the Protein Targets of Unmodified Bio‐Active Molecules with Drug Affinity Responsive Target Stability and Liquid Chromatography/Tandem Mass Spectrometry

Identifying the target proteins of bioactive small molecules is a key step in understanding mode‐of‐action of the drug and addressing the underlying mechanisms responsible for a particular phenotype. Proteomics has been successfully used to elucidate the target protein profiles of unmodified and ligand‐modified bioactive small molecules. In the latter approach, compounds can be modified via click chemistry and combined with activity‐based protein profiling. Target proteins are then enriched by performing a pull‐down with the modified ligand. Methods that utilize unmodified bioactive small molecules include the cellular thermal shift assay, thermal proteome profiling, stability of proteins from rates of oxidation, and the drug affinity responsive target stability (DARTS) determination (or read‐out). This review highlights recent proteomic approaches utilizing data‐dependent analysis and data‐independent analysis to identify target proteins by DARTS. When combined with liquid chromatography/tandem mass spectrometry, DARTS enables the identification of proteins that bind to drug molecules that leads to a conformational change in the target protein(s). In addition, an effective strategy is proposed for selecting the target protein(s) from within the pool of analyzed candidates. With additional complementary methods, the biologically relevant target proteins that bind to the small bio‐active molecules can be further validated.     

脊髓损伤扩散峰度成像参数与脊髓型颈椎病患者神经功能评分的相关性研究

目的:研究扩散峰度成像(DKI)参数与脊髓型颈椎病(CSM)患者神经功能评分的相关性及临床意义。方法:选取2018年12月至2019年6月本院收治的CSM患者37例作为研究组及健康志愿者的30例作为对照组,采用GE3.0磁共振机分别对两组人员行磁共振成像(MRI)及DKI扫描,观察其影像学特征及DKI参数的变化情况,并分析DKI参数值与临床行为评分的相关性。结果:所有研究对象的MRI图像均符合诊断要求。志愿者颈髓形态完整、信号均匀;不同年龄组颈髓平均弥散各向异性分数(FA)值、平均弥散峰度(MK)值比较差异无统计学意义(P>0.05)。根据MRI的T2加权图像上椎管受压程度及脊髓信号改变,将实验组分为A、B、C组,对照组与各实验组的MK值、FA值比较差异有统计学意义(P<0.05)。实验组FA值与mJOA评分呈显著正相关(r=0.34),与NDI评分呈负相关(r=-0.38);MK值与mJOA评分呈正相关(r=0.67),与NDI评分呈负相关(r=-0.46)。结论:DKI序列对CSM诊断具有参考较高价值,其参数与临床行为评分关系密切,能够评估早期CSM患者的脊髓损伤情况,并为诊断和治疗提供参考。     

Phylogenetic beta diversity reveals historical effects in the assemblage of the tree floras of the Ryukyu Archipelago

Quantifying the roles of historical versus contemporary constraints in determining species diversity is a central issue in island biogeography, and the phylogenetic beta diversity between islands is an essential measure specifying the influence of historical barriers on insular assemblages. In this study, using phylogenetic information for 513 tree species on 26 islands in the subtropical Ryukyu Archipelago, phylogenetic beta diversity between islands was calculated, and effects of historical factors (gaps as surrogate measures of historical barriers) and current ones (distance, area and elevation) on the phylogenetic structure of tree assemblages were examined. The pattern of phylogenetic beta diversity demonstrated that the Tokara Gap and geographical distance were consistently important for characterizing tree assemblages in the Ryukyus relative to other historical and current factors, which suggests that the Tokara Gap and distance‐limited dispersal from the two adjacent source islands have left a deep imprint on the phylogenetic structure of the current tree flora of the islands.     

Duration tenacity: A method for assessing acclimatory capacity of the Antarctic limpet, Nacella concinna

The limpet, Nacella concinna, collected from the Antarctic Peninsula (67°S), was incubated at − 0.3 °C and 2.9 °C for 9 months to test if the previously reported absence of acclimation capacity in Antarctic marine ectotherms could be due to the extended time it takes for them to adjust their physiology to a new stable state. Acclimation was tested through acute measurements of upper lethal limit and a modified measure of tenacity, that tested muscle capacity by measuring the length of time that N. concinna were able to remain attached to the substratum at different temperatures. Both measures acclimated in response to incubation to the higher temperature. Lethal limits were elevated in N. concinna incubated at 2.9 °C (8.1 ± 0.3 °C) compared to those incubated at − 0.3 °C (6.9 ± 0.4 °C). 2.9 °C incubated N. concinna also had a maximum tenacity at 2.1 °C, a higher temperature than the maximum tenacity of those incubated at − 0.3 °C, which occurred at − 1.0 °C. This study is the first to show that the Antarctic limpet can acclimate its physiology, but that it requires a greater period of time for acclimation to occur than previous studies have allowed for.     

The shape of the spatial kernel and its implications for biological invasions in patchy environments

Ecological and epidemiological invasions occur in a spatial context. We investigated how these processes correlate to the distance dependence of spread or dispersal between spatial entities such as habitat patches or epidemiological units. Distance dependence is described by a spatial kernel, characterized by its shape (kurtosis) and width (variance). We also developed a novel method to analyse and generate point-pattern landscapes based on spectral representation. This involves two measures: continuity, which is related to autocorrelation and contrast, which refers to variation in patch density. We also analysed some empirical data where our results are expected to have implications, namely distributions of trees (Quercus and Ulmus) and farms in Sweden. Through a simulation study, we found that kernel shape was not important for predicting the invasion speed in randomly distributed patches. However, the shape may be essential when the distribution of patches deviates from randomness, particularly when the contrast is high. We conclude that the speed of invasions depends on the spatial context and the effect of the spatial kernel is intertwined with the spatial structure. This implies substantial demands on the empirical data, because it requires knowledge of shape and width of the spatial kernel, and spatial structure.     

Caffeic acid phenethyl ester reduces neurovascular inflammation and protects rat brain following transient focal cerebral ischemia

Ischemic stroke is a neurovascular disease treatable by thrombolytic therapy, but the therapy has to be initiated within 3 h of the incident. This therapeutic limitation stems from the secondary injury which results mainly from oxidative stress and inflammation. A potent antioxidant/anti-inflammatory agent, caffeic acid phenethyl ester (CAPE) has potential to mitigate stroke's secondary injury, and thereby widening the therapeutic window. We observed that CAPE protected the brain in a dose-dependent manner (1-10 mg/kg body weight) and showed a wide therapeutic window (about 18 h) in a rat model of transient focal cerebral ischemia and reperfusion. The treatment also increased nitric oxide and glutathione levels, decreased lipid peroxidation and nitrotyrosine levels, and enhanced cerebral blood flow. CAPE down-regulated inflammation by blocking nuclear factor kappa B activity. The affected mediators included adhesion molecules (intercellular adhesion molecule-1 and E-selectin), cytokines (tumor necrosis factor-alpha and interleukin-1beta) and inducible nitric oxide synthase. Anti-inflammatory action of CAPE was further documented through reduction of ED1 (marker of activated macrophage/microglia) expression. The treatment inhibited apoptotic cell death by down-regulating caspase 3 and up-regulating anti-apoptotic protein Bcl-xL. Conclusively, CAPE is a promising drug candidate for ischemic stroke treatment due to its inhibition of oxidative stress and inflammation, and its clinically relevant wide therapeutic window.     

Addressing the Challenges of High‐Throughput Cancer Tissue Proteomics for Clinical Application: ProCan

The cancer tissue proteome has enormous potential as a source of novel predictive biomarkers in oncology. Progress in the development of mass spectrometry (MS)‐based tissue proteomics now presents an opportunity to exploit this by applying the strategies of comprehensive molecular profiling and big‐data analytics that are refined in other fields of ‘omics research. ProCan (ProCan is a registered trademark) is a program aiming to generate high‐quality tissue proteomic data across a broad spectrum of cancer types. It is based on data‐independent acquisition–MS proteomic analysis of annotated tissue samples sourced through collaboration with expert clinical and cancer research groups. The practical requirements of a high‐throughput translational research program have shaped the approach that ProCan is taking to address challenges in study design, sample preparation, raw data acquisition, and data analysis. The ultimate goal is to establish a large proteomics knowledge‐base that, in combination with other cancer ‘omics data, will accelerate cancer research.