Midlife Body Mass Index Trajectory and Risk of Frailty 8 Years Later in Taiwan.

OBJECTIVES: Few studies have focused on weight change and frailty, especially in Asia. This research aimed to evaluate midlife body mass index (BMI) trajectory and assess its relationship with frailty 8 years later in Taiwan. DESIGN: A prospective cohort study. SETTING AND PARTICIPANTS: Data were retrieved from the Taiwan Longitudinal Study on Aging conducted from 1999 to 2007. The analysis was restricted to respondents aged between 50 to 69 years old, who were not frail in 1999 and were alive in 2007 (n=1609). MEASUREMENTS: Frailty was defined using the Fried criteria. The group-based model of trajectory was used to estimate BMI trajectories among elderly participants. Logistic regression analysis was used to examine the association between BMI change and frailty. RESULTS: Four trajectory classes were identified and each remained stable during the 8-year follow-up. There were 316 participants (20.3%) in the low-normal weight group (baseline BMI=20.38 kg/m2), 737 participants (44.7%) in the high-normal weight group (baseline BMI=23.22 kg/m2), 449 participants (28.4%) in the overweight group (baseline BMI=26.24 kg/m2), and 107 participants (6.6%) in the obesity group (baseline BMI=30.65 kg/m2). After adjustment for confounding factors, the low-normal weight group and obesity group were associated with increased frailty compared with the high-normal weight group. CONCLUSION: Our results showed that the BMI trajectories of midlife individuals tended to be constant and those in both the low-normal weight group and obesity group had an increased risk of developing frailty in later life. Therefore, an optimal weight-targeting strategy should be considered for Asian elderly individuals.

JNK2 and IKKbeta are required for activating the innate response to viral infection.

Viral infection or double-stranded (ds) RNA induce interferons (IFN) and other cytokines. Transcription factors mediating IFN induction are known, but the signaling pathways that regulate them are less clear. We now describe two such pathways. The first pathway leading to NF-kappaB depends on the dsRNA-responsive protein kinase (PKR), which in turn activates IKB kinase (IKK) through the IKKbeta subunit. The second viral-and dsRNA-responsive pathway is PKR independent and involves Jun kinase (JNK) activation leading to stimulation of AP-1. Both IKKbeta and JNK2 are essential for efficient induction of type I IFN and other cytokines in response to viral infection or dsRNA. This study establishes a general role for these kinases in activation of innate immune responses.

Potential Alu function: regulation of the activity of double-stranded RNA-activated kinase PKR.

Cell stress, viral infection, and translational inhibition increase the abundance of human Alu RNA, suggesting that the level of these transcripts is sensitive to the translational state of the cell. To determine whether Alu RNA functions in translational homeostasis, we investigated its role in the regulation of double-stranded RNA-activated kinase PKR. We found that overexpression of Alu RNA by cotransient transfection increased the expression of a reporter construct, which is consistent with an inhibitory effect on PKR. Alu RNA formed stable, discrete complexes with PKR in vitro, bound PKR in vivo, and antagonized PKR activation both in vitro and in vivo. Alu RNAs produced by either overexpression or exposure of cells to heat shock bound PKR, whereas transiently overexpressed Alu RNA antagonized virus-induced activation of PKR in vivo. Cycloheximide treatment of cells decreased PKR activity, coincident with an increase in Alu RNA. These observations suggest that the increased levels of Alu RNAs caused by cellular exposure to different stresses regulate protein synthesis by antagonizing PKR activation. This provides a functional role for mammalian short interspersed elements, prototypical junk DNA.

Palindromic sequences preceding the terminator increase polymerase III template activity.

Four consecutive T residues in the sense strand are sufficient to terminate transcription by RNA polymerase III (pol III). Previously we observed that compared with this minimally sufficient terminator, five T residues immediately preceded by a palindromic sequence increases transcriptional expression both in vitro and in vivo, raising the question of whether a palindromic sequence has a role in pol III termination. Here we observe that site-directed mutations which eliminate the dyad symmetry of the palindromic sequence decrease transcriptional expression. Similar effects are observed whether dyad symmetry is eliminated in regions of the palindrome which are proximal or distal with respect to the terminator. Compensatory mutations at either site to restore dyad symmetry rescue transcriptional activity. These observations suggest that a higher order structure, such as a RNA hairpin, immediately preceding the terminator increases pol III transcriptional activity.

RNA polymerase III transcription repressed by Rb through its interactions with TFIIIB and TFIIIC2.

The retinoblastoma susceptibility gene product (Rb) generally represses RNA polymerase III (Pol III)-directed transcription. This implies that Rb interacts with essential transcription factors. Mutations in either the A or B subdomains in the Rb pocket interfere with Rb-mediated repression of Pol III-directed transcription, which indicates that both subdomains are directly involved in this activity. Addition of either purified TFIIIB or purified TFIIIC2 partially relieves Rb-mediated repression and restores activity to nuclear extracts that had been depleted of essential factors by binding to Rb. Pull down and coimmunoprecipitation experiments as well as functional assays indicate that Rb interacts with both TFIIIB and TFIIIC2 and that the A subdomain is primarily required for binding TFIIIB and the B subdomain for binding TFIIIC2. While Rb interacts with both factors, the A subdomain is more important than the B subdomain in directing Rb-mediated repression, and TFIIIB is the principal target of that activity.

p53 inhibits RNA polymerase III-directed transcription in a promoter-dependent manner.

Wild-type p53 represses Alu template activity in vitro and in vivo. However, upstream activating sequence elements from both the 7SL RNA gene and an Alu source gene relieve p53-mediated repression. p53 also represses the template activity of the U6 RNA gene both in vitro and in vivo but has no effect on in vitro transcription of genes encoding 5S RNA, 7SL RNA, adenovirus VAI RNA, and tRNA. The N-terminal activation domain of p53, which binds TATA-binding protein (TBP), is sufficient for repressing Alu transcription in vitro, and mutation of positions 22 and 23 in this region impairs p53-mediated repression of an Alu template both in vitro and in vivo. p53's N-terminal domain binds TFIIIB, presumably through its known interaction with TBP, and mutation of positions 22 and 23 interferes with TFIIIB binding. These results extend p53's transcriptional role to RNA polymerase III-directed templates and identify an additional level of Alu transcriptional regulation.

RNA polymerase III promoter and terminator elements affect Alu RNA expression.

Promoter elements derived from the 7SL RNA gene stimulate RNA polymerase III (Pol III) directed Alu transcription in vitro. These elements also stimulate expression of Alus transfected into 293 cells, but transcripts from these same constructs are undetectable in HeLa cells. A terminator resembling the terminator for the 7SL RNA gene has no effect on in vitro Alu template activity, but increases expression in vivo in a position independent manner. Alu transcripts generated from templates with and without this terminator have identical half-lives, indicating that this terminator stimulates expression by increasing template activity. Together, these results show that Alu expression may be regulated at multiple levels and can respond to cis-acting elements. This new found ability to express Alu transcripts by transient transfection provides an opportunity to monitor their post-transcriptional fate. Primary Alu transcripts are not extensively adenylated or deadenylated following transcription, but are short-lived compared to 118 nt scAlu RNA. In addition to Alu RNA, transfected templates encode scAlu RNA, but very high levels of Alu RNA expression does not increase the abundance of scAluRNA. ScAluRNA is not merely a transient RNA degradation product, but is instead tightly regulated by factors other than the abundance of primary transcripts.

Cell stress and translational inhibitors transiently increase the abundance of mammalian SINE transcripts.

The abundance of Alu RNA is transiently increased by heat shock in human cell lines. This effect is specific to Alu repeats among Pol III transcribed genes, since the abundance of 7SL, 7SK, 5S and U6 RNAs is essentially unaffected by heat shock. The rapid induction of Alu expression precedes the heat shock induction of mRNAs for the ubiquitin and HSP 70 heat shock genes. Heat shock mimetics also transiently induce Alu expression indicating that increased Alu expression is a general cell-stress response. Cycloheximide treatment rapidly and transiently increases the abundance of Alu RNA. Again, compared with other genes transcribed by Pol III, this increase is specific to Alu. However, as distinguished from the cell stress response, cycloheximide does not induce expression of HSP 70 and ubiquitin mRNAs. Puromycin also increases Alu expression, suggesting that this response is generally caused by translational inhibition. The response of mammalian SINEs to cell stress and translational inhibition is not limited to SINEs which are Alu homologues. Heat shock and cycloheximide each transiently induce Pol III directed expression of B1 and B2 RNAs in mouse cells and C-element RNA in rabbit cells. Together, these three species exemplify the known SINE composition of placental mammals, suggesting that mammalian SINEs are similarly regulated and may serve a common function.