Enhancing 3-hydroxypropionic acid production in Saccharomyces cerevisiae through enzyme localization within mitochondria.

Microbial 3-hydroxypropionic acid (3-HP) production can potentially replace petroleum-based production methods for acrylic acid. Here, we constructed a yeast strain that expressed enzymes related to 3-HP biosynthesis within the mitochondria. This approach aimed to enhance the 3-HP production by utilizing the mitochondrial acetyl-CoA, an important intermediate for synthesizing 3-HP. The strain that expressed 3-HP-producing enzymes in the mitochondria (YPH-mtA3HP) showed improved production of 3-HP compared to that shown by the strain expressing 3-HP-producing enzymes in the cytosol (YPH-cyA3HP). Additionally, cMCR was overexpressed, which regulates a rate-limiting reaction in synthesizing 3-HP. In this study, we aimed to further enhance 3-HP production by expressing multiple copies of cMCR in the mitochondria using the δ-integration strategy to optimize the expression level of cMCR (YPH-mtA3HPx*). The results of flask-scale cultivation showed that 3-HP production by cMCR δ-integration was significantly higher, exhibiting a yield of 160 mg/L in YPH-mtA3HP6* strain and 257 mg/L in YPH-mtA3HP22* strain. Notably, YPH-mtA3HP22*, exhibited the highest 3-HP titer, which was 3.2-fold higher than that of YPH-cyA3HP. Our results demonstrated the potential of utilizing the mitochondrial compartment within S. cerevisiae for enhancing 3-HP production.

Changes of left ventricular remodeling due to increased afterload in patients with essential hypertension.

BACKGROUND: It is unclear whether afterload mismatch occurs during the initial stage of essential hypertension (EHT). Additionally, critical left ventricular hypertrophy (LVH) between preserved and reduced systolic functions in hypertension is also unclear. Thus, we aimed to clarify these points. METHODS: Forty-five normal control subjects (NCS) and 140 EHT patients participated. EHT patients were subdivided into three groups: group I, without LVH (n = 37); group II, with LVH (n = 80); and group III, with LVH and LV heart failure (LVHF) (n = 23). Routine electrocardiographic and echocardiographic parameters, V5R/V6R ratio, relative wall thickness (RWT), LV mass (LVM) index, and peak systolic wall stress (PSWS) were measured. RESULTS: In group I, LV systolic functions were preserved despite the increase of PSWS. In group II, LVH advanced, but LV systolic functions remained normal. A negative T-wave was observed in 69% of group II and 100% of group III. A significant correlation between RWT and LVM index was seen in NCS and groups I and II (r2 = 0.545, P < 0.0001) but not in group III. Afterload mismatch occurred in group III due to the decrease in V5R/V6R ratio, the increase of LV end-diastolic dimension, and the LV systolic dysfunctions, which are caused by exhaustion of preload reserve. The boundary of the LVM index between groups II and III was approximately 180 g/m2. CONCLUSION: Afterload mismatch did not occur in group I, but it was observed in group III due to the exhaustion of preload reserve.

Baby's Online Live Database: An Open Platform for Developmental Science.

Efficient data collection in developmental studies is facing challenges due to the decreased birth rates in many regions, reproducibility problems in psychology research, and the COVID-19 pandemic. Here, we propose a novel platform for online developmental science research, the Baby's Online Live Database (BOLD), which extends the scope of the accessible participant pool, simplifies its management, and enables participant recruitment for longitudinal studies. Through BOLD, researchers can conduct online recruitment of participants preregistered to BOLD simply by specifying their attributes, such as gender and age, and direct the participants to dedicated webpages for each study. Moreover, BOLD handles participant recruitment and reward payment, thereby freeing researchers from the labor of participant management. BOLD also allows researchers the opportunity to access data that were collected from participants in previous research studies. This enables researchers to carry out longitudinal analyses at a relatively low cost. To make BOLD widely accessible, a consortium was formed within the Japan Society of Baby Science, where members from diverse research groups discussed the blueprint of this system. Once in full-scaled operation, BOLD is expected to serve as a platform for various types of online studies and facilitate international collaboration among developmental scientists in the near future.

Secondary aorto-duodenal fistula successfully treated with a novel surgical method: A case report.

INTRODUCTION: Secondary aorto-duodenal fistula (sADF) is a complication of abdominal aorta artificial blood vessel replacement that often leads to death. However, an optimal operative method has not yet been established. We describe a patient who underwent artificial blood vessel reimplantation and duodenectomy after endovascular aneurysmal repair (EVAR) with positive outcomes. PRESENTATION OF CASE: An 84-year-old man underwent artificial blood vessel replacement in 2015. In September 2016, he visited our emergency department and was diagnosed with sADF based on computed tomography. Urgent EVAR was performed, followed by duodenal segmental resection on the next day without opening the syringeal part to minimize the pollution of the operative field. Artificial blood vessel reimplantation and omental flap transposition were performed. As of 2020, about 3 years after surgery, there has been no relapse of the infection. DISCUSSION: Using our novel operative method, we can minimize exposure of the artificial blood vessel and surrounding tissue to intestinal juice and pus. We believe that this reduces the risk of postoperative artificial blood vessel reinfection. CONCLUSION: Controlling bleeding by EVAR and resection of the duodenum and artificial blood vessels as a group without opening the syringeal part can contribute to positive long-term results as this method minimizes the pollution of the duodenectomy surgical field.

[Strategy to Avoid Reoperation after Ascending Aortic Replacement for DeBakey Typeâ Acute Aortic Dissection].

When ascending aortic replacement (AAR) is performed in patients with DeBakey typeⅠ acute aortic dissection( AAD), residual false lumen of the aortic arch after AAR is a risk factor of remote aortic reoperation. We have principally considered entry resection as a surgical policy for AAD, but since 2013, for cases of re-entry in the brachiocephalic artery, reconstruction of the brachiocephalic artery has been added. We verified whether brachiocephalic artery reconstruction contributes to the thrombus closure of the false lumen of the aortic arch after the operation. Of 68 patients who underwent AAR for DeBakey typeⅠ AAD within a 12 year period, 17 had re-entry in the brachiocephalic artery on preoperative computed tomography (CT). Six patients who underwent brachiocephalic artery reconstruction were assigned to group A, and 11 patients who did not were assigned to group B. We compared the 2 groups. Aortic clamp and selective cerebral perfusion times were significantly longer in group A. By postoperative CT, false lumen of the aortic arch was not found in any case (0%) in group A, but was found in 10( 90.9%) of the 11 cases in group B( p=0.0006). The risk of residual false lumen of the aortic arch significantly decreased in group A. In conclusion, in cases where re-entry is found in the brachiocephalic artery, a false lumen often remains in the aortic arch after AAR. However, by reconstructing the dissected brachiocephalic artery, the false lumen of the aortic arch can be obliterated, which contributes to avoidance of reoperation in a chronic phase.

Short-step synthesis and chiroptical properties of polyaza[5]-[9]helicenes with blue to green-colour emission.

We have developed a two- or three-step synthesis of polyaza[5]-[9]helicenes by consecutive N-H/C-H coupling with a hypervalent iodine reagent. We further evaluated their photophysical properties, and circularly polarized luminescence properties and found that glum values increase with the number of fused rings up to 2.7 × 10-2, which is the highest value among [9]helicenes.

[Distal Stentgraft-induced New Entry Developed after Surgery for Aortic Dissection].

We experienced 2 cases with distal stentgraft-induced new entry (distal SINE) after thoracic endovascular aortic repair( TEVAR) for type B aortic dissection and 1 case after total arch replacement and frozen elephant trunk method for type A dissection. We successfully performed additional TEVAR in all cases. If the stent graft is bent at deployment, distal SINE may occur on the great curvature side of the aorta by the spring back force. Therefore, we must pay attention to appropriate device selection, and even when remodeling of the aorta is good, meticulous follow-up is necessary.

The influence of Brassica rapa var. perviridis growth conditions on the uptake and translocation of pesticides.

We cultivated Brassica rapa var. perviridis in soil mixed with four pesticides (fenobucarb, procymidone, flutolanil, and tolclofos-methyl) at different temperatures, day lengths, and soil water contents. We compared plants' uptake and translocation abilities of the pesticides as affected by growth conditions. The root concentration factor (RCF) of pesticides tended to increase with rising temperature; however, but the influence of temperature on the transpiration stream concentration factor (TSCF) differed for each pesticide. The RCFs and TSCFs of pesticides were high for short days. The soil water content had little or no effect on the uptake and translocation of pesticides. These results showed that it is necessary to consider growth conditions, especially the temperature and day length in plant uptake models for these pesticides.

Differential uptake and translocation of organic chemicals by several plant species from soil.

We performed uptake experiments with 12 different organic chemicals using 16 plant species and determined differences in the ability of plant species to take up and translocate these chemicals. There were differences among the plant species in the shoot and root concentrations of each organic chemical. The root concentration factor values increased with an increasing log of the n-octanol-water partition coefficient (log K OW) of organic chemicals. Thus, the concentrations in roots may be predicted to a certain extent because the root concentration factor values were related to the log K OW. The root-to-shoot translocation was related to the log K OW because the shoot-to-root concentration ratio decreased with an increasing log K OW; however, there was no clear relationship between the shoot concentration factor value and the log K OW, and this differed among plant species.

Listen to the outpatient: qualitative explanatory study on medical students' recognition of outpatients' narratives in combined ambulatory clerkship and peer role-play.

BACKGROUND: Understanding patients' narratives has been associated with methods of improving care that go beyond what may be regarded as a "narrow" view of scientific medicine. Medical interview training in which medical students develop understanding of the importance of patients' narratives is receiving increased attention. However, students generally receive education on patients' narratives that does not distinguish inpatients and outpatients. No studies exploring the characteristics of outpatients' narratives have been reported. We developed an educational program combining ambulatory clerkship and peer role-play using actual narratives from outpatients that students had encountered during their clerkship. These narratives were used as peer role-play scenarios in which the students acted as outpatients. This study explored what and how medical students learned about the characteristics of outpatients' narratives through this original educational program. METHODS: Participants were 70 fifth-year medical students from Nagoya University, Japan. We conducted 13 focus groups, based on a convenience sample of 11 groups in 2012, one group in 2013, and one group in 2017 (from 17 clinical groups in each year). Focus group transcripts were analyzed using the "Steps for Coding and Theorization" qualitative data analysis method. We assessed medical anthropological findings regarding narratives in a conceptual framework. RESULTS: Patients' narratives as perceived by medical students were divided into four quadrants by two axes: medical versus lived content, and objective versus subjective structure. Students recognized that outpatients' narratives mainly used a subjective structure, but were mixed and crossed each quadrant. This was described as "irreproducibility." Students also recognized that narratives of simulated patients and inpatients were mainly limited to a medical-lived content with an objective structure. These differences in narrative characteristics were recognized through students' previous interactions with simulated patients and inpatients. CONCLUSIONS: Despite some limitations, medical students learn about patients' narratives in our original educational program in a way that would be difficult to achieve through training using simulated patients or inpatients.

Effects of Gradient Coil Noise and Gradient Coil Replacement on the Reproducibility of Resting State Networks.

The stability of the MRI scanner throughout a given study is critical in minimizing hardware-induced variability in the acquired imaging data set. However, MRI scanners do malfunction at times, which could generate image artifacts and would require the replacement of a major component such as its gradient coil. In this article, we examined the effect of low intensity, randomly occurring hardware-related noise due to a faulty gradient coil on brain morphometric measures derived from T1-weighted images and resting state networks (RSNs) constructed from resting state functional MRI. We also introduced a method to detect and minimize the effect of the noise associated with a faulty gradient coil. Finally, we assessed the reproducibility of these morphometric measures and RSNs before and after gradient coil replacement. Our results showed that gradient coil noise, even at relatively low intensities, could introduce a large number of voxels exhibiting spurious significant connectivity changes in several RSNs. However, censoring the affected volumes during the analysis could minimize, if not completely eliminate, these spurious connectivity changes and could lead to reproducible RSNs even after gradient coil replacement.

Dibenzopyrrolo[1,2-a][1,8]naphthyridines: Synthesis and Structural Modification of Fluorescent L-Shaped Heteroarenes.

The L-shaped, π-extended pentacycle dibenzopyrrolo[1,2-a][1,8]naphthyridine and its derivatives were synthesized using two methods: fully intramolecular [2 + 2 + 2] cycloaddition and oxidative aromatization using substituted carbodiimide and modification of an electron-rich indole ring of an L-shaped skeleton via electrophilic reaction and cross-coupling. These L-shaped compounds emitted fluorescence in high quantum yield. The position of substituents affected the fluorescence color through two different mechanisms, π-conjugation and skeletal distortion, which caused the substituted L-shaped compounds to emit fluorescence in a variety of colors and to exhibit solvato-fluorochromism.

Japanese citizens' attitude toward end-of-life care and advance directives: A qualitative study for members of medical cooperatives.

Background: Japanese citizens are interested in choosing their own end-of-life care, but few have created their own advance directive. This study examined changes among Japanese citizens' attitudes toward end-of-life care and advance directives and explored factors that affected these attitudes. Methods: We conducted five focus groups with 48 participants in 2009 and 2010. All participants were members of health cooperatives in Tokyo. Results: We identified many barriers and reasons for creating and writing down advance directives. Experience caring for dying people and having a serious disease affected attitudes toward advance directives. Some participants changed their attitude toward end-of-life care by writing their own advance directive. Conclusions: When someone is writing advance directives, asking about his/her past experience of caring may be helpful. And learning about or filling out advance directives may help to break down resistance to using these documents.

What Is Qualitative Research?

The article is an in-depth explanation of qualitative research, an approach increasingly prevalent among today's research communities. After discussing its present spread within the health sciences, the author addresses: 1. Its definition. 2. Its characteristics, as well as its theoretical and procedural background. 3. Its procedures. 4. Differences between qualitative and quantitative approaches. 5. Mixed methods incorporating quantitative research. And in conclusion: 6. The importance of establishing an epistemological perspective in qualitative research.

Facile Two-Step Synthesis of 1,10-Phenanthroline-Derived Polyaza[7]helicenes with High Fluorescence and CPL Efficiency.

A facile two-step synthesis of aza[7]helicenes possessing a 6-5-6-6-6-5-6 skeleton from commercially available 2,9-dichloro-1,10-phenanthroline via double amination with aniline derivatives followed by hypervalent iodine reagent-mediated intramolecular double-NH/CH couplings was developed. Single-crystal X-ray analyses of the helicenes revealed unique structures, including both a significantly twisted center and planar terminals of the skeleton. The azahelicenes show high fluorescent quantum yields (Φs) under both neutral (Φ: 0.25-0.55) and acidic conditions (Φ: up to 0.80). An enantiomerically pure aza[7]helicene showed high circularly polarized luminescence (CPL) activity under both neutral and acidic conditions (glum : up to 0.009).

Integrating patients' perceptions into clinical practice guidelines for the management of rheumatoid arthritis in Japan.

OBJECTIVE: Patients' values and preferences are among the key factors that determine the strength of recommendations presented in clinical practice guidelines (CPG). The aim of this study was to summarize the integration process for patients' perceptions into the development of CPG for rheumatoid arthritis (RA) management in Japan. METHODS: We used a mixed-methods approach. Questionnaires that could be self-administered were mailed to 2222 RA patients randomly selected from the Japan Rheumatism Friendship Association (JRFA) membership list that was age- and prefecture-stratified. A focus group with five JRFA executive members was formed to verify the results of the questionnaire. RESULTS: A total of 1470 patients aged 20-79 years old returned the questionnaire. Analysis of the questionnaire data revealed that the topics selected by the CPG task force met the patients' needs. The focus group participants showed reluctance to use the term 'preference' because patients would not want to take any medications but would have to take them out of necessity. CONCLUSIONS: We confirmed that the new CPG successfully addressed clinical issues that were important to both rheumatologists and patients. Clinicians should understand patients' reluctance to take medications and explain the role of each medication well to increase adherence.

Potential application of immunoassays for simple, rapid and quantitative detections of phytoavailable neonicotinoid insecticides in cropland soils.

This study evaluated the applicability of commercially available kit-based enzyme-linked immunosorbent assay (ELISA) to simple, quick, and quantitative detection for three water-extractable (phytoavailable) neonicotinoid insecticides: dinotefuran, clothianidin, and imidacloprid in soils. ELISA showed excellent analytical sensitivity for determination, but with cross-reaction to structurally related neonicotinoid analogues, which might produce false positives. To analyze insecticides in soil samples of diverse physicochemical properties, they were extracted with water. The aqueous soil extracts were assayed directly with ELISA. No matrix interference was observed without additional dilution with water. Recovery experiments for the insecticides from aqueous soil extracts spiked at 2-10 ng/mL showed good accuracy (72-126%) and precision (<16%). Kit-based ELISAs were used to estimate soil-water distribution coefficients (Kd). Values estimated using this method showed positive correlation between organic carbon contents in soil and those for evaluated insecticides. Results indicate that the evaluated kit-based ELISA has applicability for simple, quick, and reliable detection of phytoavailable insecticides in soils and for estimating Kd values in soil.

Biodegradation of γ-hexachlorocyclohexane by transgenic hairy root cultures of Cucurbita moschata that accumulate recombinant bacterial LinA.

KEY MESSAGE: γ-HCH was successfully degraded using LinA-expressed transgenic hairy root cultures of Cucurbita moschata . Fusing an endoplasmic reticulum-targeting signal peptide to LinA was essential for stable accumulation in the hairy roots. The pesticide γ-hexachlorocyclohexane (γ-HCH) is a persistent organic pollutant (POP) that raises public health and environmental pollution concerns worldwide. Although several isolates of γ-HCH-degrading bacteria are available, inoculating them directly into γ-HCH-contaminated soil is ineffective because of the bacterial survival rate. Cucurbita species incorporate significant amounts of POPs from soils compared with other plant species. Here, we describe a novel bioremediation strategy that combines the bacterial degradation of γ-HCH and the efficient uptake of γ-HCH by Cucurbita species. We produced transgenic hairy root cultures of Cucurbita moschata that expressed recombinant bacterial linA, isolated from the bacterium Sphingobium japonicum UT26. The LinA protein was accumulated stably in the hairy root cultures by fusing an endoplasmic reticulum (ER)-targeting signal peptide to LinA. Then, we demonstrated that the cultures degraded more than 90 % of γ-HCH (1 ppm) overnight and produced the γ-HCH metabolite 1,2,4-trichlorobenzene, indicating that LinA degraded γ-HCH. These results indicate that the gene linA has high potential for phytoremediation of environmental γ-HCH.