Regulatory landscape of enhancer-mediated transcriptional activation.

Enhancers are noncoding regulatory elements that instruct spatial and temporal specificity of gene transcription in response to a variety of intrinsic and extrinsic signals during development. Although it has long been postulated that enhancers physically interact with target promoters through the formation of stable loops, recent studies have changed this static view: sequence-specific transcription factors (TFs) and coactivators are dynamically recruited to enhancers and assemble so-called transcription hubs. Dynamic assembly of transcription hubs appears to serve as a key scaffold to integrate regulatory information encoded by surrounding genome and biophysical properties of transcription machineries. In this review, we outline emerging new models of transcriptional regulation by enhancers and discuss future perspectives.

Functional coordination between transcription factor clustering and gene activity.

The prevailing view of metazoan gene regulation is that transcription is facilitated through the formation of static activator complexes at distal regulatory regions. Here, we employed quantitative single-cell live-imaging and computational analysis to provide evidence that the dynamic assembly and disassembly process of transcription factor (TF) clusters at enhancers is a major source of transcriptional bursting in developing Drosophila embryos. We further show that the regulatory connectivity between TF clustering and burst induction is highly regulated through intrinsically disordered regions (IDRs). Addition of a poly-glutamine tract to the maternal morphogen Bicoid demonstrated that extended IDR length leads to ectopic TF clustering and burst induction from its endogenous target genes, resulting in defects in body segmentation during embryogenesis. Moreover, we successfully visualized the presence of "shared" TF clusters during the co-activation of two distant genes, which provides a concrete molecular explanation for the newly proposed "topological operon" hypothesis in metazoan gene regulation.

Dynamic modulation of enhancer responsiveness by core promoter elements in living Drosophila embryos.

Regulatory interactions between enhancers and core promoters are fundamental for the temporal and spatial specificity of gene expression in development. The central role of core promoters is to initiate productive transcription in response to enhancer's activation cues. However, it has not been systematically assessed how individual core promoter elements affect the induction of transcriptional bursting by enhancers. Here, we provide evidence that each core promoter element differentially modulates functional parameters of transcriptional bursting in developing Drosophila embryos. Quantitative live imaging analysis revealed that the timing and the continuity of burst induction are common regulatory steps on which core promoter elements impact. We further show that the upstream TATA also affects the burst amplitude. On the other hand, Inr, MTE and DPE mainly contribute to the regulation of the burst frequency. Genome editing analysis of the pair-rule gene fushi tarazu revealed that the endogenous TATA and DPE are both essential for its correct expression and function during the establishment of body segments in early embryos. We suggest that core promoter elements serve as a key regulatory module in converting enhancer activity into transcription dynamics during animal development.

Oral health-related quality of life is associated with physical frailty: A cross-sectional study of Japanese community-dwelling older adults.

BACKGROUND: Physical frailty is related to adverse outcomes, and poor oral health has been linked to malnourishment. Subjective measures of oral health-related quality of life (OHRQoL) have been used as indicators of the oral health problems of older adults, and they have been associated with malnourishment. This study aimed to assess OHRQoL's association with physical frailty. METHODS: Cross-sectional study was conducted using data from the Nagasaki Islands Study that enrolled participants aged ≥60 years at Japanese national medical check-ups from 2014 to 2019. Physical frailty phenotype criteria were determined using the modified Fried frailty phenotype model. OHRQoL was assessed using the Geriatric Oral Health Assessment Index (GOHAI). Dentists conducted clinical dental examinations. Simple correlation and linear regression analyses were performed to investigate the associations of number of physical frailty phenotype criteria with GOHAI and other oral health indicators. RESULTS: Among 1341 participants with a mean age of 72 years, GOHAI score was significantly associated with number of physical frailty phenotype criteria (B = -0.01, 95% confidence interval: -0.02 to -0.01, p < 0.001). The association remained significant after adjustment for age, sex, body mass index, history of hypertension, history of diabetes mellitus, smoking status, Kessler-6 score, and number of remaining teeth. CONCLUSIONS: Oral health-related quality of life was associated with physical frailty in Japanese community-dwelling older adults.

Na+/Ca2+ exchanger mediates cold Ca2+ signaling conserved for temperature-compensated circadian rhythms.

Circadian rhythms are based on biochemical oscillations generated by clock genes/proteins, which independently evolved in animals, fungi, plants, and cyanobacteria. Temperature compensation of the oscillation speed is a common feature of the circadian clocks, but the evolutionary-conserved mechanism has been unclear. Here, we show that Na+/Ca2+ exchanger (NCX) mediates cold-responsive Ca2+ signaling important for the temperature-compensated oscillation in mammalian cells. In response to temperature decrease, NCX elevates intracellular Ca2+, which activates Ca2+/calmodulin-dependent protein kinase II and accelerates transcriptional oscillations of clock genes. The cold-responsive Ca2+ signaling is conserved among mice, Drosophila, and Arabidopsis The mammalian cellular rhythms and Drosophila behavioral rhythms were severely attenuated by NCX inhibition, indicating essential roles of NCX in both temperature compensation and autonomous oscillation. NCX also contributes to the temperature-compensated transcriptional rhythms in cyanobacterial clock. Our results suggest that NCX-mediated Ca2+ signaling is a common mechanism underlying temperature-compensated circadian rhythms both in eukaryotes and prokaryotes.

The Relationship between Dental Findings and Tongue Pressure: A Survey of 745 Community-Dwelling Adults and Elderly Persons in Japan.

INTRODUCTION: Reduced tongue pressure is one of the causes of dysphagia. The purpose of this study was to investigate the relationship between dental findings and tongue pressure, and whether prosthetic treatment prevents reduced tongue pressure. METHODS: This was a cross-sectional study. Participants were 745 community-dwelling adults and elderly persons in the Goto Islands in Nagasaki, who underwent a health checkup for residents in 2015 and 2016. Data were collected on gender; age; grip strength; hemoglobin; Creatinine (Cr); glycated hemoglobin (HbA1c); history of stroke; smoking, drinking, exercise, and walking habits; number of teeth; wearing of removable dentures; functional units of natural teeth (n-FTUs), fixed prostheses (nif-FTUs), and removable dentures (t-FTUs); and tongue pressure. The associations between each variable and tongue pressure were examined using multiple regression analysis. Next, those with 3 or fewer n-FTUs were selected, and differences in tongue pressure were compared between those with 3 or fewer nif-FTUs and those with 4 or more nif-FTUs, using a propensity score matching method. RESULTS: Male gender, weak grip strength, low HbA1c, no drinking, and a low number of teeth were independent factors significantly associated with lower tongue pressure. Among participants with 3 or fewer n-FTUs, the 43 with 4 or more nif-FTUs showed significantly higher tongue pressure than the 43 with 3 or fewer nif-FTUs after propensity score matching, although the number of t-FTUs was not associated with tongue pressure. DISCUSSION/CONCLUSION: Tooth loss was significantly associated with lower tongue pressure. It was suggested that fixed prosthesis treatment might prevent the reduction of tongue pressure, but removable dentures did not have such an effect.

Association between high psychological distress and poor oral health-related quality of life (OHQoL) in Japanese community-dwelling people: the Nagasaki Islands Study.

BACKGROUND: We investigated the association between psychological distress and oral health status/oral health-related quality of life (OHQoL) in Japanese community-dwelling people. METHODS: We conducted a cross-sectional study using data from the Nagasaki Islands Study. A total of 1183 (455 men and 728 women) has been analyzed in this study. Psychological distress was measured using the Kessler Psychological Distress Scale (K6). Oral health status was measured by dental examination. The OHQoL was measured using the General Oral Health Assessment Index (GOHAI). We defined the total score of ≥5 points on the K6 as high psychological distress (high-K6 group). RESULTS: The multiple linear regression analysis to identify the GOHAI showed that gender, K6, the total number of teeth, the number of dental caries, and visiting a dental clinic within the past 6 months significantly associated with the GOHAI. Among all of these variables, high-K6 (≥ 5) was a substantial contributing factor of the GOHAI (ß = - 0.23, 95% Cl - 2.31 to -1.41, p < 0.0001). CONCLUSIONS: It is likely that the individual with high psychological distress was strongly related to poor OHQoL even in the general population.

Involvement of glycogen metabolism in circadian control of UV resistance in cyanobacteria.

Most organisms harbor circadian clocks as endogenous timing systems in order to adapt to daily environmental changes, such as exposure to ultraviolet (UV) light. It has been hypothesized that the circadian clock evolved to prevent UV-sensitive activities, such as DNA replication and cell division, during the daytime. Indeed, circadian control of UV resistance has been reported in several eukaryotic organisms, from algae to higher organisms, although the underlying mechanisms remain unknown. Here, we demonstrate that the unicellular cyanobacterium Synechococcus elongatus PCC 7942 exhibits a circadian rhythm in resistance to UV-C and UV-B light, which is higher during subjective dawn and lower during subjective dusk. Nullification of the clock gene cluster kaiABC or the DNA-photolyase phr abolished rhythmicity with constitutively lower resistance to UV-C light, and amino acid substitutions of KaiC altered the period lengths of the UV-C resistance rhythm. In order to elucidate the molecular mechanism underlying the circadian regulation of UV-C resistance, transposon insertion mutants that alter UV-C resistance were isolated. Mutations to the master circadian output mediator genes sasA and rpaA and the glycogen degradation enzyme gene glgP abolished circadian rhythms of UV-C resistance with constitutively high UV-C resistance. Combining these results with further experiments using ATP synthesis inhibitor and strains with modified metabolic pathways, we showed that UV-C resistance is weakened by directing more metabolic flux from the glycogen degradation to catabolic pathway such as oxidative pentose phosphate pathway and glycolysis. We suggest glycogen-related metabolism in the dark affects circadian control in UV sensitivity, while the light masks this effect through the photolyase function.

Impact of H2O2 Sorption by Polymers on the Duration of Aeration in Pharmaceutical Decontamination.

As part of manufacturing a sterile drug product, we quantified the impact of H2O2 sorption by polymers on the duration of aeration in pharmaceutical decontamination. Five polymers, which are typically used as materials/parts in sterile isolators, were investigated: polyethylene, polyvinyl chloride, Silicone, polyoxymethylene (POM), and chlorosulfonated polyethylene. Experiments were performed to estimate the storage capacity and diffusion coefficients of H2O2 in the polymer. Considering these key properties of sorption/desorption, mathematical models were developed to simulate the duration of aeration to achieve the target H2O2 concentration, which is the indicator to minimize. The models were used to create a map-out of the duration given the properties of the polymers, including the five polymers. In the simulated setup, POM and Silicone were found to require prolonged aeration. Thus, when using these polymers in the isolator, the size/amount should be carefully investigated. Another practical finding was that the superiority of the polymers changed depending on the target H2O2 concentration. This result motivates an early incorporation of the product information in the isolator design, to achieve a rapid decontamination/aeration cycle.

Association of low fetuin-A levels with periodontitis in community-dwelling adults.

Fetuin-A is a liver-secreted glycoprotein isolated from fetal bovine serum. Recent reports of its several pathological functions suggest an association between fetuin-A and systemic diseases. This study therefore examined the correlation between serum fetuin-A level and periodontal status. Data from 356 middle-aged and elderly adults who underwent health examinations in Goto, Japan, during the period from 2008 through 2010 were analyzed. Systemic and periodontal measurements were recorded, and serum fetuin-A level was determined by using an enzyme-linked immunosorbent assay. Fetuin-A levels for participants with moderate to severe periodontitis were significantly lower than those for participants with no or mild periodontitis. Additionally, fetuin-A level negatively correlated with periodontal clinical attachment loss. Moderate to severe periodontitis was significantly correlated with low serum fetuin-A levels (odds ratio, 1.69; 95% confidence interval, 1.01-2.69) in logistic regression analysis. Low serum fetuin-A level was correlated with worse periodontal status and could thus potentially serve as a marker of periodontitis.

Association between human T cell leukemia virus type-1 (HTLV-1) infection and advanced periodontitis in relation to atherosclerosis among elderly Japanese: a cross-sectional study.

BACKGROUND: Human T cell leukemia virus type-1 (HTLV-1) stimulates inflammation activity. Our previous study revealed a positive association between asymptomatic HTLV-1 infection and advanced periodontitis among elderly Japanese individuals with low levels of hematopoietic activity (reflected by reticulocyte levels). Since low hematopoietic activity has been correlated with low-grade inflammation and low-grade inflammation is associated with atherosclerosis, the status of atherosclerosis could, in turn, determine the nature of this association. METHODS: To this end, a cross-sectional study of 907 elderly Japanese individuals (aged 60-99 years), who had participated in dental health check-up during the period 2016-2018, was conducted. Advanced periodontitis was defined as periodontal pocket ≥ 6.0 mm. RESULTS: Among the study population, 295 (32.5%) were found to have atherosclerosis defined as a carotid intima-media thickness (CIMT) of ≥ 1.1 mm. HTLV-1 infection was positively associated with advanced periodontitis in participants with atherosclerosis, but no significant associations were observed among the participants without atherosclerosis. The known risk factors' (including reticulocyte and CIMT) adjusted odds ratio (OR) and 95% confidence interval (CI) of advanced periodontitis were OR 2.01 and 95% CI 1.06-3.81 for participants with atherosclerosis and OR 0.61 and 95% CI 0.34-1.12 for participants without atherosclerosis. CONCLUSION: This study found a significant association between HTLV-1 infection and advanced periodontitis among elderly Japanese with atherosclerosis. However, this association is absent in individuals without atherosclerosis, suggesting that atherosclerosis might act as a determinant in the association between HTLV-1 infection and advanced periodontitis among elderly Japanese.

Association between human T cell leukemia virus 1 (HTLV-1) infection and advanced periodontitis in relation to hematopoietic activity among elderly participants: a cross-sectional study.

BACKGROUND: We reported that human T cell leukemia virus 1 (HTLV-1) infection is positively associated with atherosclerosis. Recent evidence has revealed a close association of periodontitis with atherosclerosis, endothelial dysfunction, and disruption of the microcirculation. However, the association between HTLV-1 and advanced periodontitis has not been investigated to date. Since hematopoietic activity is closely linked to endothelial maintenance activity and is known to decline with age, we hypothesized that the state of hematopoietic activity influenced the association between HTLV-1 and advanced periodontitis in elderly participants. METHODS: A cross-sectional study was performed including 822 elderly participants aged 60-99 years who participated in a dental health check-up. Advanced periodontitis was defined as a periodontal pocket ≥ 6.0 mm. Participants were classified as having low or high hematopoietic activity according to the median values of reticulocytes. RESULTS: HTLV-1 infection was positively related to advanced periodontitis among participants with lower hematopoietic activity (lower reticulocyte count), but not among participants with higher hematopoietic activity (higher reticulocyte count). The adjusted odds ratio (95% confidence interval) considering potential confounding factors was 1.92 (1.05-3.49) for participants with a lower reticulocyte count and 0.69 (0.35-1.36) for participants with a higher reticulocyte count. CONCLUSIONS: Among elderly participants, the association between HTLV-1 infection and advanced periodontitis is influenced by hematopoietic activity. Since hematopoietic activity is associated with endothelial maintenance, these findings provide an efficient tool for clarifying the underlying mechanism of the progression of periodontitis among elderly participants.

Design-oriented regression models for H2O2 decontamination processes in sterile drug product manufacturing considering rapidity and sterility.

We developed regression models for designing rapid and effective H2O2 decontamination processes in the manufacturing of sterile drug products such as injectables. Decontamination, which is typically performed by using H2O2, is a critical changeover process used to establish a sterile environment for filling products. In the process, there is a trade-off relationship between the duration of the process and the level of sterility assurance that needs to be considered in the design. Our model defines these two items as objective functions and the parameters describing the profile of H2O2 injection and the initial humidity as design variables. Our model also considers aeration, i.e., removal of H2O2 from the environment, as a part of the entire process. This design-oriented modeling considering the entire process is the novelty of the work. Experiments were performed using an industrial isolator to develop and validate a set of regression models that describe the relationship between the design variables and the objective functions. In the application of the model, Pareto-optimal conditions could be indicated given the target H2O2 concentration in aeration, which is useful for the makers of drugs and/or isolators for designing decontamination processes.

Relationship among salivary antioxidant activity, cytokines, and periodontitis: the Nagasaki Island study.

AIM: Antioxidant activities and cytokine levels in human body fluids are considered to be strongly associated with periodontitis. The aim of this study was to elucidate the relationship between salivary antioxidant activities against superoxide or hydroxyl radical, cytokines, and periodontal conditions through a community-based cross-sectional study conducted in Goto city, Japan. MATERIALS AND METHODS: Saliva samples were analysed for superoxide or hydroxyl radical scavenging activities and cytokine levels from 160 participants. We demonstrated that saliva contained superoxide and hydroxyl radical scavenging activities by using electron spin resonance with a spin-trapping agent. The concentrations of eight cytokines were measured using multiplex bead assays. RESULTS: There were significant differences in salivary superoxide or hydroxyl radical scavenging activity, and the levels of Interleukin-1ß, Interleukin-6, and Interleukin-8 between periodontitis classifications. Multivariate stepwise logistic regression model showed that salivary superoxide and hydroxyl radical scavenging activities were significantly associated with the classification of periodontitis. In addition, salivary superoxide scavenging activity was found to have significant association with all periodontal parameters using multiple linear regression analysis. CONCLUSIONS: These findings suggest that the evaluation of salivary antioxidant activities, as assessed by electron spin resonance, are associated with periodontitis and various clinical variables in community-dwelling participants (ClinicalTrials.gov number NCT01742728).

[Errata: The effects of oral health ordinances and legislation promoting dental and oral health on community-based dental and oral health organizations].

Nihon Koshu Eisei Zasshi(JAPANESE JOURNAL OF PUBLIC HEALTH), Vol. 62 (2015) No. 6 p. 294-299.

Effects of ion-releasing tooth-coating material on demineralization of bovine tooth enamel.

We compared the effect of a novel ion-releasing tooth-coating material that contained S-PRG (surface-reaction type prereacted glass-ionomer) filler to that of non-S-PRG filler and nail varnish on the demineralization of bovine enamel subsurface lesions. The demineralization process of bovine enamel was examined using quantitative light-induced fluorescence (QLF) and electron probe microanalyzer (EPMA) measurement. Ion concentrations in demineralizing solution were measured using inductively coupled plasma atomic (ICP) emission spectrometry and an ion electrode. The nail varnish group and the non-S-PRG filler group showed linear demineralization. Although the nail varnish group and the non-S-PRG filler group showed linear demineralization, the S-PRG filler group did not. Further, plane-scanning by EPMA analysis in the S-PRG filler group showed no changes in Ca ion distribution, and F ions showed peak levels on the surface of enamel specimens. Most ions in the demineralizing solution were present at higher concentrations in the S-PRG filler group than in the other two groups. In conclusion, only the S-PRG filler-containing tooth-coating material released ions and inhibited demineralization around the coating.

Association of periodontitis with carotid artery intima-media thickness and arterial stiffness in community-dwelling people in Japan: the Nagasaki Islands study.

OBJECTIVE: Recent studies have suggested an association between periodontitis and atherosclerosis; however, the relationship between periodontal status and arterial alterations should be clarified. The purpose of this study was to examine associations between periodontal status and carotid intima-media thickness (cIMT) and arterial stiffness using the cardio-ankle vascular index (CAVI) in community dwellers. METHODS: A community-based cross-sectional study of 1053 subjects ≥40 years with 10 teeth or more was conducted in Goto, Japan from 2008 to 2010. RESULTS: In a multiple linear regression analysis adjusted for age, sex, number of present teeth, and other confounders, each 1-mm increase in mean periodontal pocket depth corresponded to a 0.02-mm increase in maximal cIMT (ß = 0.018; P = 0.049) and also to a 0.1 increase in mean CAVI (ß = 0.133; P = 0.040). In addition, each 1-mm increase in the mean periodontal attachment loss corresponded to a 0.01-mm increase in maximal cIMT (ß = 0.013; P = 0.040). A multiple logistic regression analysis revealed that each 1-mm increase in mean periodontal pocket depth was associated with an increased risk of a maximal cIMT >1 mm (adjusted odds ratio [OR], 1.430; 95% confidence interval [CI], 1.067-1.918; P = 0.017) and mean CAVI of ≥8 (OR, 1.323; 95% CI, 1.003-1.743; P = 0.047). Furthermore, each 1-mm increase in mean periodontal attachment loss was associated with an increased risk of a maximal cIMT >1 mm (OR, 1.251; 95% CI, 1.032-1.516; P = 0.022). CONCLUSION: A linear, dose-dependent relationship was found between periodontal pocket depth, cIMT, and arterial stiffness.

Cessation age of breast-feeding and pacifier use is associated with persistent finger-sucking.

PURPOSE: Although some studies have reported that breast-feeding and pacifier use influence finger-sucking, few have demonstrated whether the age at cessation of breast-feeding or pacifier use and persistent finger-sucking are related. Therefore, the purpose of this study was to examine whether the age at cessation of breast-feeding and pacifier use influenced persistent finger-sucking. METHODS: A cross-sectional study of 555 36- to 47-month-olds was conducted in Nagasaki, Japan, using a questionnaire. Using the optimal cutoff point in a receiver-operating characteristic curve, the age was estimated at which cessation of pacifier use and breast-feeding had the most significant effect on persistent finger-sucking, and the estimated ages were assessed by multiple logistic regression analysis, incorporating all the questions in the questionnaire as independent variables. RESULTS: The odds ratios for persistent finger-sucking when breast-feeding was stopped at an age younger than 12 months old or when pacifier use was stopped at an age younger than 14 months old were 3.77 (95 percent confidence interval (CI)=1.97-7.22) and 8.62 (95 percent CI=2.56-29.04), respectively. CONCLUSIONS: Cessation of breast-feeding before 12 months old or pacifier use before 14 months old was associated with persistent finger-sucking.