Isoform of APOE with retained intron 3; quantitation and identification of an associated single nucleotide polymorphism

BACKGROUND: Alleles of apolipoprotein E (APOE) are the major genetic risk factor for late onset Alzheimer\u27s Disease (LOAD). Recently, an APOE splice variant that retains intron 3 (APOE-I3) was identified. To gain insight into the possible role of this isoform in LOAD, we quantified its expression in a cohort of 56 human brain specimens by using quantitative RT-PCR. RESULTS: We found that APOE-I3 generally represents a low percentage (\u3c 0.5%) of overall APOE expression. However, in one specimen, the proportion of APOE-I3 was increased about ~13 fold. This specimen was unique in the cohort for possessing the minor allele of an intron 3 single nucleotide polymorphism (SNP), rs12982192. Additionally, an allelic expression imbalance study indicated that the rs12982192 minor allele was associated with increased APOE-I3 expression. CONCLUSIONS: Overall, we interpret our results as suggesting that APOE-I3 represents a minor portion of APOE expression and that rs12982192 is associated with APOE intron 3 retention. Since the minor allele of this SNP is on the same haplotype as the minor allele of rs429358, which defines the APOE4 allele, we speculate that rs12982192 may reflect a modest loss of mRNA encoding functional APOE4

Using a Quasi-Experimental Study to Examine Program Participation and Outcomes for Older Adult Intergenerational Technology Program Participants

To extend the literature on participation in intergenerational technology programs, we conducted a quasi-experimental study consisting of senior center participants, half who took part in the University of Rhode Island Engaging Generations Cyber-Seniors (URI eGen Cyber-Seniors) Program and half who did not. Findings showed that both groups were similar on most variables; however participants did have higher education levels and more positive attitudes toward younger people. We also examined if older adult program participants improved scores on social and technological measures compared to the control group, and we found participants improved on technology use, digital competence, loneliness, and doing unpaid community service measures but not the non-participants

Reducing Ageism: Changes in Students’ Attitudes after Participation in an Intergenerational Reverse Mentoring Program

Ageism is a societal concern that greatly affects the social, emotional, physical, and mental health of older adults. One way to decrease ageist attitudes and improve the treatment of older adults is to address and improve age stereotypes among young adults. Using data from students participating in an intergenerational digital-learning program, the present study investigated change in students’ stereotypes of older adults and aging. We examined change from pre- and post-scores in student attitudes toward older adults and the type of adjectives used to describe older adults. We also analyzed responses to open-ended questions about changes in perception of older adults and aging and interest in working with older adults. Findings showed that: (1) Students’ attitudes improved following participation in the program; (2) students used fewer negative words to describe older adults following participation; (3) answers to open-ended questions demonstrated that many students improved their perceptions of older adults; and (4) many students showed increased interest in working with older adults in their future careers. Programs that reduce age stereotypes should be promoted in order to reduce young people’s harmful ageist stereotypes, ensure respectful treatment of older adults in all workplace and social situations, and increase interest in aging-related fields

Implementing an Interdisciplinary Intergenerational Program Using the Cyber Seniors ® Reverse Mentoring Model Within Higher Education

Intergenerational service-learning in higher education positively affects older adults and students, but little is known about the effectiveness of interdisciplinary, reverse mentoring programs that use technology as the medium of bringing generations together. This study describes an intergenerational service-learning program that utilizes reverse mentoring within higher education, the Engaging Generations Program, at a midsized public university in New England where students help older adults learn about technology, and students gain communication and teaching skills. In this article, we outline how the program was implemented, present quantitative data on participation outcomes for students and older adults and qualitative data from older adults, and discuss best practices. Analysis of pre/post surveys found that students\u27 attitudes toward aging improved (p \u3c 0.01) and older adults interest in technology improved (p \u3c 0.05) after program participation. Best practices identified included: multiple meetings with the same pair to deepen friendships, in-person training for student leaders, student responsibility for scheduling, tailoring sessions to each participant, student documentation of meetings, and active involvement by community partners

Improving Technology Use, Digital Competence, and Access to Community Resources Among Older Participants in the University of Rhode Island Engaging Generations Cyber-Seniors digiAGE Pilot Study

This pilot study aimed to bridge the digital divide between older and younger adults. The goal was for older people in the state to become digitally literate by engaging them in a program that provides digital devices (i.e., Apple iPads), internet connectivity (i.e., through HotSpots), and training from supervised university student mentors. This project, funded as a key policy initiative through the state’s unit on aging, specifically promoted social and economic equity by targeting participants from lower-income communities and areas hit hardest by the COVID-19 pandemic. Our university partnered with senior/community centers to recruit and support English- and Spanish-speaking adults 50 years of age and older (age range: 55-100, M=72.3, SD=8.5). For this paper, we examined changes in technology use and digital competence from the pre- to the post-survey (collected over the phone) from older participants (N=145), and we examined how the program contributed to new ways for participants to connect to community resources. Based on statistical analyses, participants improved in digital competence (pre=2.06, post=2.74), technology use (pre=1.99, post=2.70), tablet use (pre=1.53, post=4.08), and the number of purposes in which participants used technology (pre=4.09, post=5.55; p’

Regulation of the JNK3 signaling pathway during islet isolation: JNK3 and c-fos as new markers of islet quality for transplantation.

Stress conditions generated throughout pancreatic islet processing initiate the activation of pro-inflammatory pathways and beta-cell destruction. Our goal is to identify relevant and preferably beta-specific markers to assess the activation of beta-cell stress and apoptotic mechanisms, and therefore the general quality of the islet preparation prior to transplantation. Protein expression and activation were analyzed by Western blotting and kinase assays. ATP measurements were performed by a luminescence-based assay. Oxygen consumption rate (OCR) was measured based on standard protocols using fiber optic sensors. Total RNA was used for gene expression analyzes. Our results indicate that pancreas digestion initiates a potent stress response in the islets by activating two stress kinases, c-Jun N-terminal Kinase (JNK) and p38. JNK1 protein levels remained unchanged between different islet preparations and following culture. In contrast, levels of JNK3 increased after islet culture, but varied markedly, with a subset of preparations bearing low JNK3 expression. The observed changes in JNK3 protein content strongly correlated with OCR measurements as determined by the Spearman's rank correlation coefficient rho [Formula: see text] in the matching islet samples, while inversely correlating with c-fos mRNA expression [Formula: see text]. In conclusion, pancreas digestion recruits JNK and p38 kinases that are known to participate to beta-cell apoptosis. Concomitantly, the islet isolation alters JNK3 and c-fos expression, both strongly correlating with OCR. Thus, a comparative analysis of JNK3 and c-fos expression before and after culture may provide for novel markers to assess islet quality prior to transplantation. JNK3 has the advantage over all other proposed markers to be islet-specific, and thus to provide for a marker independent of non-beta cell contamination

Skeletal Muscle Differentiation Evokes Endogenous XIAP to Restrict the Apoptotic Pathway

Myotube apoptosis occurs normally during muscle development and aging but it can lead to destruction of skeletal muscle in neuromuscular diseases. Therefore, understanding how myotube apoptosis is regulated is important for developing novel strategies for treatment of muscle loss. We investigated the regulation of apoptosis in skeletal muscle and report a striking increase in resistance to apoptosis following differentiation. We find mitotic C2C12 cells (myoblast-like cells) are sensitive to cytosolic cytochrome c microinjection. However, differentiated C2C12 cells (myotube-like cells) and primary myotubes are markedly resistant. This resistance is due to endogenous X-linked inhibitor of apoptotic protein (XIAP). Importantly, the selective difference in the ability of XIAP to block myotube but not myoblast apoptosis is not due to a change in XIAP but rather a decrease in Apaf-1 expression. This decrease in Apaf-1 links XIAP to caspase activation and death. Our findings suggest that in order for myotubes to die, they may degrade XIAP, functionally inactivate XIAP or upregulate Apaf-1. Importantly, we identify a role for endogenous Smac in overcoming XIAP to allow myotube death. However, in postmitotic cardiomyocytes, where XIAP also restricts apoptosis, endogenous Smac was not capable of overcoming XIAP to cause death. These results show that as skeletal muscle differentiate, they become resistant to apoptosis because of the ability of XIAP to regulate caspase activation. The increased restriction of apoptosis in myotubes is presumably important to ensure the long term survival of these postmitotic cells as they play a vital role in the physiology of organisms

Global analysis of gene expression in NGF-deprived sympathetic neurons identifies molecular pathways associated with cell death

Developing sympathetic neurons depend on nerve growth factor (NGF) for survival and die by apoptosis after NGF withdrawal. This process requires de novo gene expression but only a small number of genes induced by NGF deprivation have been identified so far, either by a candidate gene approach or in mRNA differential display experiments. This is partly because it is difficult to obtain large numbers of sympathetic neurons for in vitro studies. Here, we describe for the first time, how advances in gene microarray technology have allowed us to investigate the expression of all known genes in sympathetic neurons cultured in the presence and absence of NGF