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1.
Sci Rep ; 13(1): 17396, 2023 10 13.
Article in English | MEDLINE | ID: mdl-37833342

ABSTRACT

In the field of applied microbiology, reproducibility and experimental variability are important factors that influence both basic research as well as process development for industrial applications. Experimental reproducibility and accuracy depend not only on culture conditions such as temperature and aeration but also on raw materials and procedures used for media preparation. The M9 minimal medium is one of the most common synthetic media for culturing Escherichia coli and other bacteria. This synthetic medium can be used to observe and evaluate the physiological activity of microbes under minimal nutritional requirements and determine the limiting factor for the desired phenotype. Although one of the advantages using the M9 medium is that its composition can be modulated, it is difficult to control presence of trace components and impurities from the reagents for preparing this medium. Herein, we showed that trace ingredients present in the reagents used for M9 media preparation affect the bacterial physiological activities (e.g., cell growth, substrate consumption, and byproduct formation). Additionally, we systematically identified the trace ingredient that influenced phenotypic differences. Our results showed that the selection of reagents and accuracy during reagent preparation is important for experimental reproducibility in the field of bio-engineering and systems biology focused on the systematic and continuous development of biomolecular systems (e.g., biorefinery, metabolic engineering, and synthetic biology).


Subject(s)
Escherichia coli , Phosphates , Escherichia coli/genetics , Reproducibility of Results , Culture Media/chemistry
2.
Sci Rep ; 13(1): 13608, 2023 09 04.
Article in English | MEDLINE | ID: mdl-37666852

ABSTRACT

Bioprocess optimization using mathematical models is prevalent, yet the discrepancy between model predictions and actual processes, known as process-model mismatch (PMM), remains a significant challenge. This study proposes a novel hybrid control system called the hybrid in silico/in-cell controller (HISICC) to address PMM by combining model-based optimization (in silico feedforward controller) with feedback controllers utilizing synthetic genetic circuits integrated into cells (in-cell feedback controller). We demonstrated the efficacy of HISICC using two engineered Escherichia coli strains, TA1415 and TA2445, previously developed for isopropanol (IPA) production. TA1415 contains a metabolic toggle switch (MTS) to manage the competition between cell growth and IPA production for intracellular acetyl-CoA by responding to external input of isopropyl ß-D-1-thiogalactopyranoside (IPTG). TA2445, in addition to the MTS, has a genetic circuit that detects cell density to autonomously activate MTS. The combination of TA2445 with an in silico controller exemplifies HISICC implementation. We constructed mathematical models to optimize IPTG input values for both strains based on the two-compartment model and validated these models using experimental data of the IPA production process. Using these models, we evaluated the robustness of HISICC against PMM by comparing IPA yields with two strains in simulations assuming various magnitudes of PMM in cell growth rates. The results indicate that the in-cell feedback controller in TA2445 effectively compensates for PMM by modifying MTS activation timing. In conclusion, the HISICC system presents a promising solution to the PMM problem in bioprocess engineering, paving the way for more efficient and reliable optimization of microbial bioprocesses.


Subject(s)
2-Propanol , Escherichia coli , Isopropyl Thiogalactoside , Acetyl Coenzyme A , Cell Cycle , Cell Proliferation , Escherichia coli/genetics
3.
BMC Med Educ ; 23(1): 419, 2023 Jun 07.
Article in English | MEDLINE | ID: mdl-37286967

ABSTRACT

BACKGROUND: The effectiveness of peer learning has been recognized and discussed by many scholars, and implemented in the formal curriculums of medical schools internationally. However, there is a general dearth of studies in measuring the objective outcomes in learning. METHODS: We investigated the objective effect of near-peer learning on tutee's emotions and its equivalence within the formal curriculum of a clinical reasoning Problem Based Learning session in a Japanese medical school. Fourth-year medical students were assigned to the group tutored by 6th-year students or by faculties. The positive activating emotion, positive deactivating emotion, negative activating emotion, negative deactivating emotion, Neutral emotion were measured using the Japanese version of the Medical Emotion Scale (J-MES), and self-efficacy scores were also assessed. We calculated the mean differences of these variables between the faculty and the peer tutor groups and were statistically analyzed the equivalence of these scores. The equivalence margin was defined as a score of 0.4 for J-MES and 10.0 for the self-efficacy score, respectively. RESULTS: Of the 143 eligible participant students, 90 were allocated to the peer tutor group and 53 were allocated to the faculty group. There was no significant difference between the groups. The 95% confidence interval of the mean score difference for positive activating emotions (-0.22 to 0.15), positive deactivating emotions (-0.35 to 0.18), negative activating emotions (-0.20 to 0.22), negative deactivating emotions (-0.20 to 0.23), and self-efficacy (-6.83 to 5.04) were withing the predetermined equivalence margins for emotion scores, meaning that equivalence was confirmed for these variables. CONCLUSIONS: Emotional outcomes were equivalent between near-peer PBL sessions and faculty-led sessions. This comparative measurement of the emotional outcomes in near-peer learning contributes to understanding PBL in the field of medical education.


Subject(s)
Education, Medical, Undergraduate , Students, Medical , Humans , Problem-Based Learning , Students, Medical/psychology , Curriculum , Emotions
4.
Children (Basel) ; 10(1)2023 Jan 12.
Article in English | MEDLINE | ID: mdl-36670704

ABSTRACT

The COVID-19 pandemic has greatly changed medical education, and medical trainees' self-regulation has become more emphasized. In Japan, the concept of self-regulated learning has not been fully applied in health profession education due to a lack of effective measurement tools. We aimed to validate the translated Japanese version of the Motivated Strategies for Learning Questionnaire in the context of Problem-Based Learning (J-MSLQ-PBL). The questionnaire employs a seven-point Likert-type scale with 81 items and is categorized into two sections: motivation and learning strategies. An exploratory factor analysis (EFA) was conducted by using Promax rotation to examine the factor structure of the scale, using the collected data from 112 Japanese medical students. Factor extraction was based on a scree plot investigation, and an item was accepted when the factor loading was ≥0.40. In the motivation section, the extracted factors from the EFA were well aligned with the subscales of the original MSLQ, including "Self-Efficacy for Learning and Performance", "Task Value", "Self-Efficacy for Learning and Performance", "Test Anxiety", "Extrinsic Goal Orientation", and "Intrinsic Goal Orientation". In the learning strategies, the extracted factors poorly matched the structure of the original subscales. This discrepancy could be explained by insufficient translation, the limited sample size from a single medical school, or cross-cultural differences in learning strategies between Western and Japanese medical students. Only the motivation part of the J-MSLQ-PBL should be implemented to measure the competency elements of self-regulated learning in Japan.

5.
Health Place ; 79: 102952, 2023 01.
Article in English | MEDLINE | ID: mdl-36535074

ABSTRACT

We prospectively investigated the association between acceptable travel distances (i.e., walking and cycling) and the incidence of functional disability and mortality among older Japanese adults. A baseline survey was conducted in 2013 among 7618 individuals aged ≥65 years in Kasama City, Japan, and they were tracked through the city's database until 2021. Acceptable travel distances were assessed using a questionnaire. Outcomes (i.e., functional disability and mortality) were gathered as binary (incident or not) with survival time. A multivariable-adjusted Cox proportional-hazards model indicated that shorter acceptable walking and cycling distances were markedly linked with higher risks of functional disability and mortality. In conclusion, acceptable distances of ≤500 m for walking and ≤1 km for cycling were associated with higher risks of functional disability and mortality among older adults.


Subject(s)
East Asian People , Walking , Humans , Aged , Follow-Up Studies , Proportional Hazards Models , Incidence
6.
Metabolites ; 12(9)2022 Aug 28.
Article in English | MEDLINE | ID: mdl-36144211

ABSTRACT

Pre-column fluorescent derivatization has been used for the fast quantification of amino acids using high-performance liquid chromatography (HPLC) systems. However, it generally requires an offline in-vial derivatization process with multiple derivatization reagents. The offline derivatization requires the same number of reaction vials as the number of sample vials for use as a reaction chamber for the derivatization reaction in an autosampler. Therefore, the number of samples analyzed per batch using the pre-column derivatization method is halved. To benefit from the pre-column derivatization method, we transformed the derivatization process from an offline chamber process to an online in-needle process (in-needle Pre-column Derivatization for Amino acids Quantification; iPDAQ). Fluorescent derivatization in the injection needle obviated the need for vacant vials as reaction chambers. Consequently, the throughput per batch improved up to two times, and the consumption of derivatization reagents was reduced to less than one-tenth of that in the conventional vial method. We demonstrated to separate and quantify the amino acids in various biological samples. Herein, we presented a novel HPLC-based amino acid quantification method that enables the continuous analysis of a large number of samples. The iPDAQ facilitates accurate amino acid quantification due to the automation of derivatization and achieves improvement in the throughput and reduction of analysis labor.

7.
MedEdPublish (2016) ; 12: 25, 2022.
Article in English | MEDLINE | ID: mdl-37082001

ABSTRACT

Background: In response to globalism, many East-Asian countries now include a Medical English course in their undergraduate medical education syllabus. The purpose of this study was to explore the relationship between the related attribute factors of students' motivation to learn medical English through an online modality. Methods: Of 134 eligible fourth-year medical students who participated in an Online Medical English course at a Japanese medical school, 105 were enrolled in this single cohort study. The participants completed pre- and post-course surveys regarding their motivation during the course, including perceived academic control and task value, and their assignment scores. A structural equation model was used to examine the hypothesized relationship of constructs, based on control-value theory. Results: The model showed a good fit for the data (χ 2[df=7] = 1.821, p=0.969, CFI = 1.000, RMSEA < 0.001, SRMR < 0.05, GFI = 0.993, AGFI = 0.980). The latent variables of the perceived course achievement related to the observed variables of academic control and task value scale scores, and negatively predicted willingness for self-study after course completion. In addition, the preference of English as the course language negatively predicted willingness for self-study of medical English. Conclusion: Choice of English as the language of instruction and perceived high course achievement negatively predicted students' motivation for further English self-study after the class. The importance of incorporating the perspective of lifelong learning into the teaching of medical English was recognized.

8.
J Biosci Bioeng ; 133(1): 46-55, 2022 Jan.
Article in English | MEDLINE | ID: mdl-34620543

ABSTRACT

The production of chemicals and fuels from renewable resources using engineered microbes is an attractive alternative for current fossil-dependent industries. Metabolic engineering has contributed to pathway engineering for the production of chemicals and fuels by various microorganisms. Recently, dynamic metabolic engineering harnessing synthetic biological tools has become a next-generation strategy in this field. The dynamic regulation of metabolic flux during fermentation optimizes metabolic states according to each fermentation stage such as cell growth phase and compound production phase. However, it is necessary to repeat the evaluation and redesign of the dynamic regulation system to achieve the practical use of engineered microbes. In this study, we performed quantitative metabolome analysis to investigate the effects of dynamic metabolic flux regulation on engineered Escherichia coli for γ-amino butyrate (GABA) fermentation. We prepared a stable isotope-labeled internal standard mixture (SILIS) for the stable isotope dilution method (SIDM), a mass spectrometry-based quantitative metabolome analysis method. We found multiple candidate bottlenecks for GABA production. Some metabolic reactions in the GABA production pathway should be engineered for further improvement in the direct GABA fermentation with dynamic metabolic engineering strategy.


Subject(s)
Metabolic Engineering , Metabolomics , Escherichia coli/genetics , Fermentation , Isotopes , Metabolome
9.
J Biosci Bioeng ; 133(1): 56-63, 2022 Jan.
Article in English | MEDLINE | ID: mdl-34674961

ABSTRACT

Pyruvate is a key intermediate that is involved in various synthetic metabolic pathways for microbial chemical and fuel production. It is widely used in the food, chemical, and pharmaceutical industries. However, the microbial production of pyruvate and its derivatives compete with microbial cell growth, as pyruvate is an important metabolic intermediate that serves as a hub for various endogenous metabolic pathways, including gluconeogenesis, amino acid synthesis, TCA cycle, and fatty acid biosynthesis. To achieve a more efficient bioprocess for the production of pyruvate and its derivatives, it is necessary to reduce the metabolic imbalance between cell growth and target chemical production. For this purpose, we devised a dynamic metabolic engineering strategy within an Escherichia coli model, in which a metabolic toggle switch (MTS) was employed to redirect metabolic flux from the endogenous pathway toward the target synthetic pathway. Through a combination of TCA cycle interruption through MTS and reduction of pyruvate consumption in endogenous pathways, we achieved a drastic improvement (163 mM, 26-fold) in pyruvate production. In addition, we demonstrated the redirection of metabolic flux from excess pyruvate toward isobutanol production. The final isobutanol production titer of the strain harboring MTS was 26% improved compared with that of the control strain.


Subject(s)
Escherichia coli Proteins , Metabolic Engineering , Escherichia coli/genetics , Escherichia coli/metabolism , Escherichia coli Proteins/genetics , Fermentation , Pyruvic Acid
10.
Article in English | MEDLINE | ID: mdl-34831701

ABSTRACT

This study aimed to examine the factors related to participation in volunteer-managed preventive care exercises by focusing on the distance to exercise facilities and interpersonal social networks. A postal mail survey was conducted in 2013 in Kasama City in a rural region of Japan. Older adults (aged ≥ 65 years) who were living independently (n = 16,870) were targeted. Potential participants who were aware of silver-rehabili taisou exercise (SRTE) and/or square-stepping exercise (SSE) were included in the analysis (n = 4005). A multiple logistic regression analysis revealed that social and environmental factors were associated with participation in SRTE and SSE. After adjusting for confounding variables, exercise participation was negatively associated with an extensive distance from an exercise facility in both sexes for SRTE and SSE. Among women, participation in SRTE was negatively associated with weak interpersonal social networks (odds ratio (OR) = 0.57), and participation in SRTE and SSE was negatively associated with being a car passenger (SRTE, OR = 0.76; SSE, OR = 0.60). However, there were no significant interactions between sex and social and environmental factors. Our findings suggest the importance of considering location and transportation to promote participation in preventive care exercise.


Subject(s)
Exercise , Volunteers , Aged , Cross-Sectional Studies , Female , Humans , Japan , Male , Social Networking , Social Participation
11.
Medicine (Baltimore) ; 100(37): e27269, 2021 Sep 17.
Article in English | MEDLINE | ID: mdl-34664881

ABSTRACT

ABSTRACT: The mortality of the bath-related cardiac arrest (BRCA) is extremely high. While air temperature is reported to be associated with the BRCA occurrence, it is unclear whether daily minimum temperatures or the difference between maximum and minimum air temperatures influences BRCA occurrence the most.A retrospective cohort study of adult patients was conducted between January 2015 and February 2020 at Hirosaki University Hospital Emergency Department. The following data were collected: age, sex, day of cardiac arrest event, location of the event, initial cardiac rhythm, presence of return of spontaneous circulation, and overall mortality (status at 1 month after cardiac arrest event). Based on the day of the event and the location in which the event occurred, daily minimum and maximum temperatures were obtained from the Japan Meteorological Agency database.A total of 215 eligible cardiac arrest cases were identified, including 25 cases of BRCA. Comparing BRCA and non-BRCA, initial shockable cardiac rhythm (4.0% vs 44.7%), presence of return of spontaneous circulation (8.0% vs 34.7%), and overall mortality (96.0% vs 71.6%) differed significantly (P < .05 each). Daily minimum and maximum temperatures showed no significant relationships with BRCA or non-BRCA. Daily minimum temperature was a risk factor of BRCA occurrence after adjusting for age and temperature difference (risk ratio, 0.937; 95% confidence interval, 0.882-0.995).Daily minimum temperature represents a potential risk factor for BRCA occurrence.


Subject(s)
Baths/statistics & numerical data , Out-of-Hospital Cardiac Arrest/mortality , Temperature , Adult , Aged , Aged, 80 and over , Baths/adverse effects , Cohort Studies , Female , Humans , Japan/epidemiology , Male , Middle Aged , Out-of-Hospital Cardiac Arrest/complications , Out-of-Hospital Cardiac Arrest/epidemiology , Retrospective Studies
12.
Biochem Biophys Rep ; 27: 101097, 2021 Sep.
Article in English | MEDLINE | ID: mdl-34401533

ABSTRACT

ß-Hydroxy-ß-methylbutyrate (HMB) is an intermediate in the metabolism of the branched-chain amino acid leucine. HMB has several demonstrated effects on skeletal muscle function, some of which are contradictory. In addition, the effect of exogenous HMB intake on the levels of intermediate metabolites is not known. Therefore, we investigated changes in HMB metabolites after oral HMB administration in mice. First, ICR mice were treated with either distilled water or HMB (0.215 g/10 mL/kg). Sampling was performed at 0, 1, 6, 12, and 24 h after administration. Next, ICR mice were given distilled water or HMB (0.215 g/10 mL/kg/d) for 10 d. Mice given HMB shown a significant increase in liver ß-methylcrotonyl-CoA and increased ß-hydroxybutyrate in plasma and the gastrocnemius muscle 1 h after HMB administration. Mice administered HMB for 10 d showed significantly decreased food intake and body weight; however, the relative weight of the gastrocnemius muscle was significantly increased. These results may be attributed to an increase in ß-hydroxybutyrate resulting from exogenous HMB, since ß-hydroxybutyrate inhibits food intake and suppresses skeletal muscle catabolism. In conclusion, ß-hydroxybutyrate, a metabolite of HMB, was found to play an important role in the function of HMB.

13.
ACS Synth Biol ; 10(6): 1384-1393, 2021 06 18.
Article in English | MEDLINE | ID: mdl-34106678

ABSTRACT

Dynamic metabolic engineering that harnesses synthetic biological tools is a next-generation strategy for microbial chemical and fuel production. We previously reported a synthetic quorum sensing system combined with a metabolic toggle switch (QS-MTS) in E. coli. It autonomously redirected endogenous metabolic flux toward the synthetic metabolic pathway and improved biofuel production. However, its functions and effects on host metabolism were attenuated by induction timing delay. Here, we redesigned the QS-MTS to stabilize QS signaling efficiency and metabolic regulation. We performed a metabolome analysis to clarify the effects of QS-MTS redesign on host metabolism. We compared the contributions of conventional and redesigned QS-MTS to fed-batch fermentation. The redesigned QS-MTS was more conducive than the conventional QS-MTS to long-term processes such as fed-batch fermentation. Here, we present a circuit redesign for metabolic flux control based on dynamic characteristic evaluation and metabolome analysis.


Subject(s)
Escherichia coli/metabolism , Metabolic Engineering/methods , Metabolome/genetics , Quorum Sensing/genetics , Signal Transduction/genetics , Batch Cell Culture Techniques/methods , Biofuels , Escherichia coli/genetics , Fermentation , Gene Expression , Gene Expression Regulation, Bacterial , Gene Regulatory Networks , Metabolomics/methods , Microorganisms, Genetically-Modified , Time Factors
14.
Metabolites ; 11(4)2021 Mar 30.
Article in English | MEDLINE | ID: mdl-33808182

ABSTRACT

Calibration-Curve-Locking Databases (CCLDs) have been constructed for automatic compound search and semi-quantitative screening by gas chromatography/mass spectrometry (GC/MS) in several fields. CCLD felicitates the semi-quantification of target compounds without calibration curve preparation because it contains the retention time (RT), calibration curves, and electron ionization (EI) mass spectra, which are obtained under stable apparatus conditions. Despite its usefulness, there is no CCLD for metabolomics. Herein, we developed a novel CCLD and semi-quantification framework for GC/MS-based metabolomics. All analytes were subjected to GC/MS after derivatization under stable apparatus conditions using (1) target tuning, (2) RT locking technique, and (3) automatic derivatization and injection by a robotic platform. The RTs and EI mass spectra were obtained from an existing authorized database. A quantifier ion and one or two qualifier ions were selected for each target metabolite. The calibration curves were obtained as plots of the peak area ratio of the target compounds to an internal standard versus the target compound concentration. These data were registered in a database as a novel CCLD. We examined the applicability of CCLD for analyzing human plasma, resulting in time-saving and labor-saving semi-qualitative screening without the need for standard substances.

15.
Bioresour Technol ; 330: 125002, 2021 Jun.
Article in English | MEDLINE | ID: mdl-33770731

ABSTRACT

This study demonstrates the metabolic alteration of Methylococcus capsulatus (Bath), a representative bacterium among methanotrophs, in microbial gas-phase reactions. For comparative metabolome analysis, a bioreactor was designed to be capable of supplying gaseous substrates and liquid nutrients continuously. Methane degradation by M. capsulatus (Bath) was more efficient in a gas-phase reaction operated in the bioreactor than in an aqueous phase reaction operated in a batch reactor. Metabolome analysis revealed remarkable alterations in the metabolism of cells in the gas-phase reaction; in particular, pyruvate, 2-ketoglutarate, some amino acids, xanthine, and hypoxanthine were accumulated, whereas 2,6-diaminopimelate was decreased. Based on the results of metabolome analysis, cells in the gas-phase reaction seemed to alter their metabolism to reduce the excess ATP and NADH generated upon increased availability of methane and oxygen. Our findings will facilitate the development of efficient processes for methane-based bioproduction with low energy consumption.


Subject(s)
Biochemical Phenomena , Methylococcus capsulatus , Bioreactors , Methane , Methylococcus capsulatus/metabolism , Oxygen , Oxygenases/metabolism
16.
PLoS One ; 15(11): e0242308, 2020.
Article in English | MEDLINE | ID: mdl-33201904

ABSTRACT

Pulmonary diseases, including chronic obstructive pulmonary disease (COPD), are major chronic diseases that result in decreased pulmonary function. Relationships between body composition and pulmonary function have been reported. However, few epidemiological studies have used the visceral fat area (VFA) to measure body composition. This study aimed to examine the relationship between body composition and pulmonary function. A cross-sectional study was conducted between 2015 and 2016, using data obtained from 1,287 residents aged between 19 and 91 years living in the Iwaki area of Hirosaki City, a rural region in Aomori Prefecture, Japan. Pulmonary function was evaluated using the forced vital capacity (FVC) as a percentage of the predicted value (predicted FVC%) and the ratio of forced expiratory volume in one second (FEV1) to FVC. The measurements for evaluating body composition included the body fat percentage (BFP) of the whole body and trunk, skeletal muscle index (SMI), body mass index (BMI), VFA, waist circumference (WC) at the navel level, and waist-to-hip ratio (WHR). To adjust for potential confounders, Spearman's partial correlation analysis was used to examine the relationship between the measurements of body composition and pulmonary function. There were significant correlations between the predicted FVC% and the following parameters: BFP (whole body and trunk) in younger males; SMI in older males; WC, VFA, BMI, and SMI in younger females; and BFP (whole body and trunk) and VFA in older females. Contrastingly, WC and VFA in younger males and WC in younger females were correlated with the FEV1/FVC ratio. VFA was correlated with the FEV1/FVC ratio in younger males and predicted FVC% in older females. These findings suggest that visceral fat accumulation may increase the development of obstructive pulmonary disease in young males and accelerate the decline of pulmonary function (predicted FVC%) in older females.


Subject(s)
Body Composition/physiology , Vital Capacity/physiology , Adult , Aged , Aged, 80 and over , Body Mass Index , Cross-Sectional Studies , Female , Humans , Insulin Resistance , Interleukin-6/analysis , Intra-Abdominal Fat/physiology , Japan , Male , Middle Aged , Muscle, Skeletal/physiology , Waist Circumference , Waist-Hip Ratio , Young Adult
17.
Nihon Koshu Eisei Zasshi ; 67(9): 573-581, 2020.
Article in Japanese | MEDLINE | ID: mdl-33041283

ABSTRACT

Objectives The present study examined factors that affect learning about dental caries and periodontal disease before or after graduation or completion of training for dietitians and registered dietitians.Methods A questionnaire survey was conducted with members of the Aomori Prefectural Dietetic Association between October and November 2019, and 276 participants were included in the analysis. A multiple logistic regression analysis was performed to assess the characteristics associated with learning about dental caries and periodontal disease; odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated. The following independent variables were assessed: age (<30, 30-39, 40-49, 50-59, or ≥60 years), license (dietitian or registered dietitian), and occupational field (medical institution [nutrition management or food service], administrative agency, nursing care insurance facility, school-related work, training instructor in training facility for dietitians and registered dietitians, or other).Results The multiple logistic regression analysis showed that dietitians and registered dietitians who had learned about dental caries and periodontal disease during training tended to be below the age of 40 years and have a registered dietitian license (dental caries: OR=2.79, 95% CI=1.08-7.24; periodontal disease: OR=6.51, 95% CI=1.71-24.84). Furthermore, dietitians and registered dietitians who had learned about dental caries and periodontal disease after graduation or training completion tended to be over the age of 40, have studied at a training facility (dental caries: OR=3.21, 95% CI=1.65-6.27; periodontal disease: OR=3.06, 95% CI=1.32-7.12), and be employed in the field of school-related work (dental caries: OR=4.23, 95% CI=1.03-17.27; periodontal disease: OR=5.56, 95% CI=1.15-26.98).Conclusions To facilitate increased cooperation among practitioners in the fields of nutrition and dental health, necessary opportunities for learning about dental caries and periodontal disease alongside experts should be provided to those who do not have a registered dietitian license and have not studied at training facilities.


Subject(s)
Dental Caries , Dietetics/education , Education, Continuing/methods , Health Education, Dental , Learning , Nutritionists/education , Periodontal Diseases , Adult , Curriculum , Female , Humans , Middle Aged , Surveys and Questionnaires
18.
Metab Eng ; 57: 23-30, 2020 01.
Article in English | MEDLINE | ID: mdl-31377410

ABSTRACT

Genetic manipulation in cyanobacteria enables the direct production of valuable chemicals from carbon dioxide. However, there are still very few reports of the production of highly effective photosynthetic chemicals. Several synthetic metabolic pathways (e.g., isopropanol, acetone, isoprene, and fatty acids) have been constructed by branching from acetyl-CoA and malonyl-CoA, which are key intermediates for photosynthetic chemical production downstream of pyruvate decarboxylation. Recent reports of the absolute determination of cellular metabolites in Synechococcus elongatus PCC 7942 have shown that its acetyl-CoA levels corresponded to about one hundredth of the pyruvate levels. In short, one of the reasons for lower photosynthetic chemical production from acetyl-CoA and malonyl-CoA was the smaller flux to acetyl-CoA. Pyruvate decarboxylation is a primary pathway for acetyl-CoA synthesis from pyruvate and is mainly catalyzed by the pyruvate dehydrogenase complex (PDHc). In this study, we tried to enhance the flux toward acetyl-CoA from pyruvate by overexpressing PDH genes and, thus, catalyzing the conversion of pyruvate to acetyl-CoA via NADH generation. The overexpression of PDH genes cloned from S. elongatus PCC 7942 significantly increased PDHc enzymatic activity and intracellular acetyl-CoA levels in the crude cell extract. Although growth defects were observed in overexpressing strains of PDH genes, the combinational overexpression of PDH genes with the synthetic metabolic pathway for acetate or isopropanol resulted in about 7-fold to 9-fold improvement in its production titer, respectively (9.9 mM, 594.5 mg/L acetate, 4.9 mM, 294.5 mg/L isopropanol). PDH genes overexpression would, therefore, be useful not only for the production of these model chemicals, but also for the production of other chemicals that require acetyl-CoA as a key precursor.


Subject(s)
Acetyl Coenzyme A , Bacterial Proteins , Metabolic Networks and Pathways , Photosynthesis , Pyruvate Dehydrogenase Complex , Synechococcus , 2-Propanol/metabolism , Acetates/metabolism , Acetyl Coenzyme A/genetics , Acetyl Coenzyme A/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Metabolic Engineering , Pyruvate Dehydrogenase Complex/genetics , Pyruvate Dehydrogenase Complex/metabolism , Synechococcus/genetics , Synechococcus/metabolism
19.
Metab Eng ; 55: 268-275, 2019 09.
Article in English | MEDLINE | ID: mdl-31401244

ABSTRACT

Synthetic microbial consortia consisting of microorganisms with different synthetic genetic circuits or divided synthetic metabolic pathway components can exert functions that are beyond the capacities of single microorganisms. However, few consortia of microorganisms with different synthetic genetic circuits have been developed. We designed and constructed a synthetic microbial consortium composed of an enzyme-producing strain and a target chemical-producing strain using Escherichia coli for chemical production with efficient saccharification. The enzyme-producing strain harbored a synthetic genetic circuit to produce beta-glucosidase, which converts cellobiose to glucose, destroys itself via the lytic genes, and release the enzyme when the desired cell density is reached. The target chemical-producing strain was programmed by a synthetic genetic circuit to express enzymes in the synthetic metabolic pathway for isopropanol production when the enzyme-producing strain grows until release of the enzyme. Our results demonstrate the benefits of synthetic microbial consortia with distributed tasks for effective chemical production from biomass.


Subject(s)
2-Propanol/metabolism , Cellobiose , Escherichia coli , Glucose , Microbial Consortia , Microorganisms, Genetically-Modified , Cellobiose/genetics , Cellobiose/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Glucose/genetics , Glucose/metabolism , Microorganisms, Genetically-Modified/genetics , Microorganisms, Genetically-Modified/metabolism
20.
J Phys Ther Sci ; 29(10): 1737-1741, 2017 Oct.
Article in English | MEDLINE | ID: mdl-29184280

ABSTRACT

[Purpose] There is a relationship between physical and cognitive functions; therefore, impairment of physical function would mean cognitive decline. This study aimed to investigate the association between change in physical and cognitive functions. [Subjects and Methods] Participants were 169 healthy community-dwelling older adults who attend the survey after three years from baseline (mean age, 72.4 ± 4.8 years). Grip strength, one-leg standing balance, five-times-sit-to-stand test, timed up and go, 5-m habitual walk, and a peg-moving task were used to evaluate physical performance. Five cognitive function tests were used to assess attention, memory, visuospatial function, verbal fluency, and reasoning. Cognitive function was defined as the cumulative score of these tests. [Results] At baseline, five-times-sit-to-stand test, timed up and go, and hand dexterity were independently associated with cognitive function. In longitudinal analyses, changes in habitual walking speed and hand dexterity were significantly associated with change in cognitive function. [Conclusion] Deterioration of specific physical function, such as hand dexterity and walking ability, may be associated with progression of cognitive decline. Decreasing extent of daily functions, such as hand dexterity and walking ability, can be useful indices to grasp changes in cognitive function.

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