Accumulation of “Old Proteins” and the Critical Need for MS‐based Protein Turnover Measurements in Aging and Longevity

Aging and age‐related diseases are accompanied by proteome remodeling and progressive declines in cellular machinery required to maintain protein homeostasis (proteostasis), such as autophagy, ubiquitin‐mediated degradation, and protein synthesis. While many studies have focused on capturing changes in proteostasis, the identification of proteins that evade these cellular processes has recently emerged as an approach to studying the aging proteome. With advances in proteomic technology, it is possible to monitor protein half‐lives and protein turnover at the level of individual proteins in vivo. For large‐scale studies, these technologies typically include the use of stable isotope labeling coupled with MS and comprehensive assessment of protein turnover rates. Protein turnover studies have revealed groups of highly relevant long‐lived proteins (LLPs), such as the nuclear pore complexes, extracellular matrix proteins, and protein aggregates. Here, the role of LLPs during aging and age‐related diseases and the methods used to identify and quantify their changes are reviewed. The methods available to conduct studies of protein turnover, used in combination with traditional proteomic methods, will enable the field to perform studies in a systems biology context, as changes in proteostasis may not be revealed in studies that solely measure differential protein abundances.     

A Comprehensive Analysis of Symmetric Arginine Dimethylation in Colorectal Cancer Tissues Using Immunoaffinity Enrichment and Mass Spectrometry

Protein arginine methyltransferase 5 (PRMT5) is a major enzyme responsible for generating monomethyl and symmetric dimethyl arginine in proteins. PRMT5 is essential for cell viability and development, and its overexpression is observed in a variety of cancers. In the present study, it is found that levels of PRMT5 protein and symmetric arginine dimethylation in colorectal cancer (CRC) tissues are increased compared to those in adjacent noncancerous tissues. Using immunoaffinity enrichment of methylated peptides combined with high‐resolution mass spectrometry, a total of 147 symmetric dimethyl‐arginine (SDMA) sites in 94 proteins are identified, many of which are RNA binding proteins and enzymes. Quantitative analysis comparing CRC and normal tissues reveals significant increase in the symmetric dimethylation of 70 arginine sites in 46 proteins and a decrease in that of four arginine sites in four proteins. Among the 94 proteins identified in this study, it is confirmed that KH‐type splicing regulatory protein is a target of PRMT5 and highly expressed in CRC tissues compared to noncancerous tissues. This study is the first comprehensive analysis of symmetric arginine dimethylation using clinical samples and extends the number of known in vivo SDMA sites. The data obtained are available via ProteomeXchange with the identifier PXD015653.     

Enhancing Data Reliability in TOMAHAQ for Large‐Scale Protein Quantification

The analytical scale of most mass‐spectrometry‐based targeted proteomics assays is usually limited by assay performance and instrument utilization. A recently introduced method, called triggered by offset, multiplexed, accurate mass, high resolution, and absolute quantitation (TOMAHAQ), combines both peptide and sample multiplexing to simultaneously improve analytical scale and quantitative performance. In the present work, critical technical requirements and data analysis considerations for successful implementation of the TOMAHAQ technique based on the study of a total of 185 target peptides across over 200 clinical plasma samples are discussed. Importantly, it is observed that significant interference originate from the TMTzero reporter ion used for the synthetic trigger peptides. This interference is not expected because only TMT10plex reporter ions from the target peptides should be observed under typical TOMAHAQ conditions. In order to unlock the great promise of the technique for high throughput quantification, here a post‐acquisition data correction strategy to deconvolute the reporter ion superposition and recover reliable data is proposed.     

Investigating the Effects of Chemical Gradients on Performance and Reliability within Perovskite Solar Cells with TOF‐SIMS

Time‐of‐flight secondary‐ion mass spectrometry (TOF‐SIMS), a powerful analytical technique sensitive to all components of perovskite solar cell (PSC) materials, can differentiate between the various organic species within a PSC absorber or a complete device stack. The ability to probe chemical gradients through the depth of a device (both organic and inorganic), with down to 100 nm lateral resolution, can lead to unique insights into the relationships between chemistry in the absorber bulk, at grain boundaries, and at interfaces as well as how they relate to changes in performance and/or stability. In this review, the technique is described; then, from the literature, several examples of how TOF‐SIMS have been used to provide unique insight into PSC absorbers and devices are covered. Finally, the common artifacts that can be introduced if the data are improperly collected, as well as methods to mitigate these artifacts are discussed.     

喉罩通气下七氟醚全凭吸入麻醉在小儿先天性心脏病介入手术的临床麻醉效果

目的:探讨喉罩通气下七氟醚全凭吸入麻醉在小儿先天性心脏病介入手术的临床麻醉效果。方法:选取2017年4月～2019年5月期间我院收治的行先天性心脏病介入手术患儿98例,根据随机数字表法将其分为对照组(n=49)和研究组(n=49)。对照组给予氯胺酮诱导,全凭丙泊酚维持,面罩吸氧;研究组给予全凭七氟醚诱导、维持,喉罩通气。比较两组患儿麻醉前(T0)、切皮前(T1)、切皮后1 min(T2)、切皮后30 min(T3)、术后(T4)的血流动力学指标[平均动脉压(MAP)、心率(HR)]及应激反应指标[血糖、皮质醇],记录两组患儿手术时间、麻醉诱导时间、术后苏醒时间等围术期指标情况。记录两组围术期不良反应发生情况。结果:研究组手术时间、麻醉诱导时间、术后苏醒时间均短于对照组(P0.05)。两组T0时间点血糖、MAP、皮质醇、HR比较差异无统计学意义(P0.05);对照组T1～T4时间点MAP、血糖、皮质醇、HR均较T0升高(P0.05);研究组T1～T4时间点血糖、MAP、皮质醇、HR与T0时间点比较无差异(P0.05);研究组T1～T4时间点血糖、MAP、皮质醇、HR低于对照组(P0.05)。两组不良反应发生率比较无差异(P0.05)。结论:小儿先天性心脏病介入手术中应用喉罩通气下七氟醚全凭吸入麻醉,诱导迅速且安全、术后苏醒快、手术时间短,可有效维持血流动力学稳定,减少应激反应。     

1-6岁儿童维生素D与年龄、性别、季节及体质量指数的关系研究

目的:探讨1-6岁儿童维生素D与年龄、性别、季节及体质量指数(BMI)的关系,为临床有效指导维生素D的补充提供参考依据。方法:选取2000年1月～2019年1月于我院接受体检的1～6岁儿童816例作为研究对象。采用电化学发光法检测所有研究对象的血清25-羟维生素D[25(OH)D]水平,并分析血清25(OH)D水平与儿童年龄、性别、BMI以及季节的关系。结果:816例儿童维生素D营养不足和缺乏人数占比为14.83%。1～3岁儿童血清25(OH)D水平高于3～6岁儿童(P0.05)。男童血清25(OH)D水平高于女童(P0.05)。肥胖儿童血清25(OH)D水平低于正常与超重儿童,且超重儿童血清25(OH)D水平低于正常儿童(均P0.05)。春、夏季儿童血清25(OH)D水平均高于秋、冬季儿童(均P0.05)。结论:1～6岁儿童的维生素D营养状况不容乐观,随着年龄的增长儿童血清25(OH)D水平显著降低,且男童高于女童,春、夏季高于秋、冬季。临床工作可通过增加其户外活动,继而达到改善维生素D营养状况的目的。     

Domestication is associated with differential expression of pikeperch egg proteins involved in metabolism,immune response and protein folding

Domestication is a condition in which the breeding, care and feeding of animals are, at least in part, controlled by humans. Information regarding the changes in the protein composition of eggs in response to domestication is very limited. Such data are prerequisite for improvements in the reproduction of domesticated fish. The aim of this study was to examine the impact of domestication on the proteome of pikeperch eggs using two-dimensional differential in-gel electrophoresis. We analysed high-quality eggs from domesticated and wild pikeperch fish to reveal proteins that were presumably only related to the domestication process and not to the quality of eggs. Here, we show that domestication has a profound impact on the protein profile of pikeperch eggs. We identified 66 differentially abundant protein spots, including 27 spots that were more abundant in wild-caught pikeperch eggs and 39 spots that were enriched in eggs collected from domesticated females. Eggs originating from wild-caught females showed higher expression levels of proteins involved in folding, apoptotic process, purine metabolism and immune response, whereas eggs of domesticated females showed higher expression levels of proteins that participated mainly in metabolism. The changes in metabolic proteins in eggs from domesticated females can reflect the adaptation of pikeperch to commercial diets, which have profoundly distinct compositions compared with natural diets. The decrease in the abundance of proteins related to immune response in eggs from the domesticated population suggests that domestication may lead to disturbances in defence mechanisms. In turn, the lower abundance of heat shock proteins in eggs of domesticated fish may indicate their adaptation to stable farming conditions and reduced environmental stressors or their better tolerance of stress from breeding. The proteins identified in this study can increase our knowledge concerning the mechanism of the pikeperch domestication process.     

Unchartered waters: Climate change likely to intensify infectious disease outbreaks causing mass mortality events in marine mammals

Infectious disease emergence has increased significantly over the last 30 years, with mass mortality events (MMEs) associated with epizootics becoming increasingly common. Factors influencing these events have been widely studied in terrestrial systems, but remain relatively unexplored in marine mammals. Infectious disease‐induced MMEs (ID MMEs) have not been reported ubiquitously among marine mammal species, indicating that intrinsic (host) and/or extrinsic (environmental) ecological factors may influence this heterogeneity. We assess the occurrence of ID MMEs (1955–2018) across extant marine mammals (n = 129) in relation to key life‐history characteristics (sociality, trophic level, habitat breadth) and environmental variables (season, sea surface temperature [SST] anomalies, El Niño occurrence). Our results show that ID MMEs have been reported in 14% of marine mammal species (95% CI 9%–21%), with 72% (n = 36; 95% CI 56%–84%) of these events caused predominantly by viruses, primarily morbillivirus and influenza A. Bacterial pathogens caused 25% (95% CI 14%–41%) of MMEs, with only one being the result of a protozoan pathogen. Overall, virus‐induced MMEs involved a greater number of fatalities per event compared to other pathogens. No association was detected between the occurrence of ID MMEs and host characteristics, such as sociality or trophic level, but ID MMEs did occur more frequently in semiaquatic species (pinnipeds) compared to obligate ocean dwellers (cetaceans; χ² = 9.6, p = .002). In contrast, extrinsic factors significantly influenced ID MMEs, with seasonality linked to frequency (χ² = 19.85, p = .0002) and severity of these events, and global yearly SST anomalies positively correlated with their temporal occurrence (Z = 3.43, p = 2.7e‐04). No significant association was identified between El Niño and ID MME occurrence (Z = 0.28, p = .81). With climate change forecasted to increase SSTs and the frequency of extreme seasonal weather events, epizootics causing MMEs are likely to intensify with significant consequences for marine mammal survival.