Influence of effortful swallow on pharyngeal pressure: evaluation using a high-resolution manometry.

To evaluate the effect of effortful swallow on pharyngeal pressure while swallowing saliva and water using a novel high-resolution manometry (HRM) system

Gesture imitation performance in community-dwelling older people: assessment of a gesture imitation task in the screening and diagnosis of mild cognitive impairment and dementia

Takasaki A., Hashimoto M., Fukuhara R., et al. Gesture imitation performance in community-dwelling older people: assessment of a gesture imitation task in the screening and diagnosis of mild cognitive impairment and dementia. Psychogeriatrics 24, 404 (2024); https://doi.org/10.1111/psyg.13086.Background: Gesture imitation, a simple tool for assessing visuospatial/visuoconstructive functions, is reportedly useful for screening and diagnosing dementia. However, gesture imitation performance in healthy older adults is largely unknown, as are the factors associated with lower performance. To address these unknowns, we examined the gesture imitation performance of a large number of community-dwelling older adults aged ≥65 years in Arao City, Kumamoto Prefecture (southern Japan). Methods: The examiner presented the participants with eight gesture patterns and considered it a success if they could imitate them within 10 s. The success rate of each gesture imitation was calculated for three diagnostic groups: cognitively normal (CN) (n = 1184), mild cognitive impairment (MCI) (n = 237), and dementia (n = 47). Next, we reorganised the original gesture imitation battery by combining six selected gestures with the following scoring method: if the participants successfully imitated the gestures, immediately or within 5 s, two points were assigned. If they succeeded within 5–10 s, one point was assigned. The sensitivity and specificity of the battery were investigated to detect the dementia and MCI groups. Factors associated with gesture imitation battery scores were examined. Results: Except one complex gesture, the success rate of imitation in the CN group was high, approximately 90%. The sensitivity and specificity of the gesture imitation battery for discriminating between the dementia and CN groups and between the MCI and CN groups were 70%/88%, and 45%/75%, respectively. Ageing, male sex, and a diagnosis of dementia or MCI were associated with lower scores on the gesture imitation battery. Conclusion: Gesture imitation tasks alone may not be sufficient to detect MCI. However, by combining gestures with set time limits, gesture imitation tasks can be a low-burden and effective method for detecting dementia, even in community medicine, such as during health check-ups

U2 snRNP Is Required for Expression of the 3′ End of Genes

<div><p>Pre-mRNA in eukaryotes is subjected to mRNA processing, which includes capping, polyadenylation, and splicing. Transcription and mRNA processing are coupled, and this coupling stimulates mRNA processing; however, the effects of mRNA processing on transcription are not fully understood. In this study, we found that inhibition of U2 snRNP by a splicing inhibitor, spliceostatin A (SSA), or by an antisense oligonucleotide to U2 snRNA, caused gene-specific 3′-end down-regulation. Removal of SSA from the culture media restored expression of the 3′ ends of genes, suggesting that U2 snRNP is required for expression of the 3′ end of genes. Finally, we found that SSA treatment caused accumulation of Pol II near the 5′ end of 3′-end down regulated genes, such as <i>CDK6</i>, <i>SMEK2</i> and <i>EGFR</i>, indicating that SSA treatment led to transcription elongation arrest on these genes. These findings suggest that U2 snRNP is important for production of full length mRNA probably through regulation of transcription elongation, and that a novel checkpoint mechanism prevents pre-mRNA from accumulating as a result of splicing deficiencies, and thereby prevents production of aberrant proteins that might be translated from pre-mRNAs through the arrest of transcription elongation.</p></div

Pol II accumulates near the 5′ end of genes.

<p>(A, C, E, G) The locations of primers used in this assay are shown under the gene structures. (B, D, F, H) HeLa cells were treated with 30 ng/ml of SSA for 2 hours, and the distributions of Pol II were analyzed by chromatin immunoprecipitation (ChIP). The amount of immunoprecipitated DNA fragments was measured by quantitative PCR (upper panels). The ratios between the percent input of SSA-treated cells (SSA) and methanol-treated cells (MeOH) were calculated (MeOH  = 1) (lower panels). Error bars indicate s.d. (n = 4). Statistical significance was investigated by the t-test (*: p<0.05; **: p<0.01).</p

SSA treatment causes splicing inhibition and 3′-end down-regulation.

<p>(A, B, C) HeLa cells were treated with the indicated concentrations of SSA for 4 hours, and RNAs were labeled during transcription with EU between 3 and 4 hours after the addition of SSA (A). Labeled RNAs were analyzed by quantitative RT-PCR to measure the amounts of spliced and unspliced mRNA. Relative splicing activity was defined as the ratio of spliced to unspliced mRNA (methanol-treated cells [Ctrl]  = 100%) (B). The levels of the 5′ and 3′ ends of <i>CDK6</i> (Exon 2 and Exon 8, respectively) were measured by quantitative RT-PCR (C, upper panel), and the ratio between the 3′ and 5′ levels was calculated (methanol-treated cells [Ctrl]  = 100%) (C, lower panel). (D, E, F) HeLa cells were treated with 100 ng/ml of SSA, and RNAs were labeled during transcription with EU for 1 hour as indicated (D). Labeled RNAs were analyzed as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098015#pone-0098015-g002" target="_blank">Figure 2B and 2C</a> (methanol-treated cells [Ctrl]  = 100%) (E, F). (G, H, I) HeLa cells were treated with 100 ng/ml of SSA for 2 hours, and then washed with fresh medium. Cells were cultivated in fresh medium, and RNAs were labeled during transcription with EU for 1 hour as indicated (G). Labeled RNAs were analyzed as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098015#pone-0098015-g002" target="_blank">Figure 2B and 2C</a> (methanol-treated cells [Ctrl]  = 100%) (H, I). Error bars indicate s.d. (n = 3). Statistical significance was investigated by the t-test (*: p<0.05; **: p<0.01; ***: p<0.001).</p

3′-end down-regulation by SSA treatment is observed in exosome deficient cells.

<p>(A, B) HeLa cells were transfected with Control or <i>RRP4</i> siRNA (20 nM), and then cultured for 48 hours after transfection. Cells were then treated with the indicated concentration of SSA for 4 hours, and RNAs were labeled during transcription with EU between 3 and 4 hours after the addition of SSA. (A) Half of the transfected cells were analyzed by Western blotting to measure the level of RRP4 protein. Molecular weights are indicated to the left of the gels. (B) RNA samples were purified from the other half of the cells, and labeled RNAs were analyzed by quantitative RT-PCR. The ratios between the 3′ and 5′ ends of <i>CDK6</i>, <i>SMEK2</i>, <i>VEGFA</i> and <i>EGFR</i> were calculated (methanol-treated and control siRNA-transfected cells  = 100%). Error bars indicate s.d. (n = 3). Statistical significance (RRP4 siRNA, 0 ng/ml SSA vs. RRP4 siRNA, 30 ng/ml SSA) was investigated by the t-test (**: p<0.01; ***: p<0.001).</p