Optimization of Orthogonal Translation Systems Enhances Access to the Human Phosphoproteome

Protein phosphorylation is a ubiquitous post-translational modification that governs signaling cascades and protein-protein interactions. Orthogonal translation systems, in which an orthogonal aminoacyl-tRNA synthetase and tRNA pair have been repurposed for insertion of a non-standard amino acid, have been developed for co-translational insertion of phospho-amino acids. Here, we systematically developed variants of phosphoserine and phosphothreonine orthogonal translation systems to minimize cellular toxicity and maximize translational fidelity. We characterized novel tRNA constructs to enhance cellular fitness and change decoding functionality. Multiple genetic background strains were developed to enhance the incorporation of phospho-amino acids into recombinant proteins. Utilizing large-scale DNA synthesis, we were able to produce serine, threonine, and tyrosine libraries representative of the entire known human phosphoproteome. By pairing our libraries with a bimolecular fluorescence complementation assay, we were able to characterize novel phosphorylation-dependent protein-protein interactions for phosphoserine, phosphothreonine, and phosphotyrosine. Genetically encoded phosphothreonine enabled the discovery of a new activation mechanism for the protein kinase CHK2 and proteome-wide surveys of its target substrates. Finally, we developed Hi-P+ to couple kinase substrate discovery with phosphorylation-dependent protein-protein interactions. This work enables kinase-specific, proteome-wide surveys of multi-level phosphorylation-dependent interactions with phosphosite resolution

Keyboarding for Elementary Students

This package contains four components designed to help elementary or business education teachers teach keyboarding to elementary students. The keyboarding program described in these materials consists of three phases: Phase I--learn the keyboard; basic function keys; technique and confidence; Phase II--review the keyboard and technique; language arts application; introduction to composition and proofreading; and Phase III--quick review of keyboard and technique; creating; editing; and printing; and computer terminology. This package contains (1) an inservice guide for business teachers and elementary teachers that provides suggestions for introducing and implementing a keyboarding program; (2) a teacher\u27s guide; with a day-by-day lesson plan for a six-week course in keyboarding that could be used in Phase I and for review; (3) Appendix A; with supplemental materials such as suggestions for implementation; sample letters to parents; visual keyboard; sample name card; activity suggestions; activity sheets; technique check list; glossary of computer terms; certificate and good work awards; and picture of a computer bug ; and (4) Appendix B; which contains references for software; textbooks; and a bibliography of articles that relate to keyboarding. (KC

A small erythropoietin derived non-hematopoietic peptide reduces cardiac inflammation, attenuates age associated declines in heart function and prolongs healthspan

BackgroundAging is associated with increased levels of reactive oxygen species and inflammation that disrupt proteostasis and mitochondrial function and leads to organism-wide frailty later in life. ARA290 (cibinetide), an 11-aa non-hematopoietic peptide sequence within the cardioprotective domain of erythropoietin, mediates tissue protection by reducing inflammation and fibrosis. Age-associated cardiac inflammation is linked to structural and functional changes in the heart, including mitochondrial dysfunction, impaired proteostasis, hypertrophic cardiac remodeling, and contractile dysfunction. Can ARA290 ameliorate these age-associated cardiac changes and the severity of frailty in advanced age?MethodsWe conducted an integrated longitudinal (n = 48) and cross-sectional (n = 144) 15 months randomized controlled trial in which 18-month-old Fischer 344 x Brown Norway rats were randomly assigned to either receive chronic ARA290 treatment or saline. Serial echocardiography, tail blood pressure and body weight were evaluated repeatedly at 4-month intervals. A frailty index was calculated at the final timepoint (33 months of age). Tissues were harvested at 4-month intervals to define inflammatory markers and left ventricular tissue remodeling. Mitochondrial and myocardial cell health was assessed in isolated left ventricular myocytes. Kaplan–Meier survival curves were established. Mixed ANOVA tests and linear mixed regression analysis were employed to determine the effects of age, treatment, and age-treatment interactions.ResultsChronic ARA290 treatment mitigated age-related increases in the cardiac non-myocyte to myocyte ratio, infiltrating leukocytes and monocytes, pro-inflammatory cytokines, total NF-κB, and p-NF-κB. Additionally, ARA290 treatment enhanced cardiomyocyte autophagy flux and reduced cellular accumulation of lipofuscin. The cardiomyocyte mitochondrial permeability transition pore response to oxidant stress was desensitized following chronic ARA290 treatment. Concurrently, ARA290 significantly blunted the age-associated elevation in blood pressure and preserved the LV ejection fraction. Finally, ARA290 preserved body weight and significantly reduced other markers of organism-wide frailty at the end of life.ConclusionAdministration of ARA290 reduces cell and tissue inflammation, mitigates structural and functional changes within the cardiovascular system leading to amelioration of frailty and preserved healthspan

Physiological Correlates of Volunteering

We review research on physiological correlates of volunteering, a neglected but promising research field. Some of these correlates seem to be causal factors influencing volunteering. Volunteers tend to have better physical health, both self-reported and expert-assessed, better mental health, and perform better on cognitive tasks. Research thus far has rarely examined neurological, neurochemical, hormonal, and genetic correlates of volunteering to any significant extent, especially controlling for other factors as potential confounds. Evolutionary theory and behavioral genetic research suggest the importance of such physiological factors in humans. Basically, many aspects of social relationships and social activities have effects on health (e.g., Newman and Roberts 2013; Uchino 2004), as the widely used biopsychosocial (BPS) model suggests (Institute of Medicine 2001). Studies of formal volunteering (FV), charitable giving, and altruistic behavior suggest that physiological characteristics are related to volunteering, including specific genes (such as oxytocin receptor [OXTR] genes, Arginine vasopressin receptor [AVPR] genes, dopamine D4 receptor [DRD4] genes, and 5-HTTLPR). We recommend that future research on physiological factors be extended to non-Western populations, focusing specifically on volunteering, and differentiating between different forms and types of volunteering and civic participation

Retirement and encore adulthood: the new later life course

Retirement became an institutionalized, primarily male, later life-course transition by the middle of the twentieth century; an expected, routine exit from a lifetime of paid work that was also seen as the passage to old age. However, large-scale social forces (globalization, aging populations, women's labor force participation, reduced social protections) have upended conventional expectations about retirement and the later life course. The period roughly between ages 55 and 75. years has arguably become a new life course stage, an 'encore' to conventional adulthood occurring typically after the career- and family-building years, but prior to the infirmities associated with old age

Stress Proliferation? Precarity and Work–Family Conflict at the Intersection of Gender and Household Income

We theorize a stress proliferation process whereby the stress of job precarity translates into the stress of work-to-family conflict (WFC). We test whether this process differs by gender and household income. Using four cross-sectional waves of the General Social Survey (N = 2,340), we find a positive association between job insecurity and WFC for women but not men. Examined by household income levels, the association is found only for respondents in the lowest income tercile. Furthermore, gender intersects with household income to shape the stress proliferation process. While the insecurity–WFC relationship holds for women across all household income levels, for men this relationship shifts from positive for men in the lowest income tercile to negative for men in the highest income tercile. Our findings suggest that entrenched gendered expectations around work and family may lead women (regardless of household income) and lower-class men to be most vulnerable to stress proliferation