Protective effect of UV-irradiated red perilla (Perilla frutescens (L.) Britton) on carbon tetrachloride-induced liver injury in mice.

UV-irradiated red perilla demonstrated promising protective effects against carbon tetrachloride-induced liver injury in mice. UV exposure significantly enhanced the accumulation of rosmarinic acid, malonylshisonin, and shisonin in red perilla, and increased 1,1-diphenyl-2-picrylhydrazyl radical scavenging capacity. The hepatoprotective effect of UV-irradiated red perilla may be attributed to the high level of its polyphenolic compounds, which exhibit antioxidant activity.

Comparative whole-genome and proteomics analyses of the next seed bank and the original master seed bank of MucoRice-CTB 51A line, a rice-based oral cholera vaccine.

BACKGROUND: We have previously developed a rice-based oral vaccine against cholera diarrhea, MucoRice-CTB. Using Agrobacterium-mediated co-transformation, we produced the selection marker-free MucoRice-CTB line 51A, which has three copies of the cholera toxin B subunit (CTB) gene and two copies of an RNAi cassette inserted into the rice genome. We determined the sequence and location of the transgenes on rice chromosomes 3 and 12. The expression of alpha-amylase/trypsin inhibitor, a major allergen protein in rice, is lower in this line than in wild-type rice. Line 51A was self-pollinated for five generations to fix the transgenes, and the seeds of the sixth generation produced by T5 plants were defined as the master seed bank (MSB). T6 plants were grown from part of the MSB seeds and were self-pollinated to produce T7 seeds (next seed bank; NSB). NSB was examined and its whole genome and proteome were compared with those of MSB. RESULTS: We re-sequenced the transgenes of NSB and MSB and confirmed the positions of the three CTB genes inserted into chromosomes 3 and 12. The DNA sequences of the transgenes were identical between NSB and MSB. Using whole-genome sequencing, we compared the genome sequences of three NSB with three MSB samples, and evaluated the effects of SNPs and genomic structural variants by clustering. No functionally important mutations (SNPs, translocations, deletions, or inversions of genic regions on chromosomes) between NSB and MSB samples were detected. Analysis of salt-soluble proteins from NSB and MSB samples by shot-gun MS/MS detected no considerable differences in protein abundance. No difference in the expression pattern of storage proteins and CTB in mature seeds of NSB and MSB was detected by immuno-fluorescence microscopy. CONCLUSIONS: All analyses revealed no considerable differences between NSB and MSB samples. Therefore, NSB can be used to replace MSB in the near future.

Subquinocin, a small molecule inhibitor of CYLD and USP-family deubiquitinating enzymes, promotes NF-κB signaling.

The tumor suppressor CYLD negatively regulates polyubiquitination-dependent cellular signaling such as nuclear factor (NF)-κB signaling. In addition to CYLD, multiple deubiquitinating enzymes (DUBs) are also involved in the regulation of this signaling pathway, and distinct role of CYLD is yet to be clarified. Here, we identified a small chemical named Subquinocin that inhibited the DUB activity of recombinant CYLD using a wheat cell-free protein synthesis and an AlphaScreen technology. In cells, Subquinocin increased the polyubiquitination of NEMO and RIP1 and enhanced NF-κB activation. Modeling and mutation analyses indicated that Subquinocin interacted with Y940 in CYLD, which locates close to catalytic center of CYLD, and is conserved among the USP-family DUBs. Further biochemical evaluation revealed that Subquinocin inhibited USP-family DUBs, but not other family DUBs including OTU. Although Subquinocin showed a broad specificity toward USP-family DUBs, the inhibitory effect of Subquinocin on NF-κB signaling was negligible in CYLD-KO cells, indicating that CYLD is a major target of Subquinocin on the suppression of NF-κB signaling. In conclusion, Subquinocin identified here is a useful tool to analyze the signal transduction mediated by USP-family DUBs.

Monitoring the Growth and Yield of Fruit Vegetables in a Greenhouse Using a Three-Dimensional Scanner.

Monitoring the growth of fruit vegetables is essential for the automation of cultivation management, and harvest. The objective of this study is to demonstrate that the current sensor technology can monitor the growth and yield of fruit vegetables such as tomato, cucumber, and paprika. We estimated leaf area, leaf area index (LAI), and plant height using coordinates of polygon vertices from plant and canopy surface models constructed using a three-dimensional (3D) scanner. A significant correlation was observed between the measured and estimated leaf area, LAI, and plant height (R2 > 0.8, except for tomato LAI). The canopy structure of each fruit vegetable was predicted by integrating the estimated leaf area at each height of the canopy surface models. A linear relationship was observed between the measured total leaf area and the total dry weight of each fruit vegetable; thus, the dry weight of the plant can be predicted using the estimated leaf area. The fruit weights of tomato and paprika were estimated using the fruit solid model constructed by the fruit point cloud data extracted using the RGB value. A significant correlation was observed between the measured and estimated fruit weights (tomato: R2 = 0.739, paprika: R2 = 0.888). Therefore, it was possible to estimate the growth parameters (leaf area, plant height, canopy structure, and yield) of different fruit vegetables non-destructively using a 3D scanner.

HTLV-1 Tax Induces Formation of the Active Macromolecular IKK Complex by Generating Lys63- and Met1-Linked Hybrid Polyubiquitin Chains.

The Tax protein of human T-cell leukemia virus type 1 (HTLV-1) is crucial for the development of adult T-cell leukemia (ATL), a highly malignant CD4+ T cell neoplasm. Among the multiple aberrant Tax-induced effects on cellular processes, persistent activation of transcription factor NF-κB, which is activated only transiently upon physiological stimulation, is essential for leukemogenesis. We and others have shown that Tax induces activation of the IκB kinase (IKK) complex, which is a critical step in NF-κB activation, by generating Lys63-linked polyubiquitin chains. However, the molecular mechanism underlying Tax-induced IKK activation is controversial and not fully understood. Here, we demonstrate that Tax recruits linear (Met1-linked) ubiquitin chain assembly complex (LUBAC) to the IKK complex and that Tax fails to induce IKK activation in cells that lack LUBAC activity. Mass spectrometric analyses revealed that both Lys63-linked and Met1-linked polyubiquitin chains are associated with the IKK complex. Furthermore, treatment of the IKK-associated polyubiquitin chains with Met1-linked-chain-specific deubiquitinase (OTULIN) resulted in the reduction of high molecular weight polyubiquitin chains and the generation of short Lys63-linked ubiquitin chains, indicating that Tax can induce the generation of Lys63- and Met1-linked hybrid polyubiquitin chains. We also demonstrate that Tax induces formation of the active macromolecular IKK complex and that the blocking of Tax-induced polyubiquitin chain synthesis inhibited formation of the macromolecular complex. Taken together, these results lead us to propose a novel model in which the hybrid-chain-dependent oligomerization of the IKK complex triggered by Tax leads to trans-autophosphorylation-mediated IKK activation.

Decreased linear ubiquitination of NEMO and FADD on apoptosis with caspase-mediated cleavage of HOIP.

NF-κB is crucial to regulate immune and inflammatory responses and cell survival. LUBAC generates a linear ubiquitin chain and activates NF-κB through ubiquitin ligase (E3) activity in the HOIP subunit. Here, we show that HOIP is predominantly cleaved by caspase at Asp390 upon apoptosis, and that is subjected to proteasomal degradation. We identified that FADD, as well as NEMO, is a substrate for LUBAC. Although the C-terminal fragment of HOIP retains NF-κB activity, linear ubiquitination of NEMO and FADD decreases upon apoptosis. Moreover, the N-terminal fragment of HOIP binds with deubiquitinases, such as OTULIN and CYLD-SPATA2. These results indicate that caspase-mediated cleavage of HOIP divides critical functional regions of HOIP, and that this regulates linear (de)ubiquitination of substrates upon apoptosis.

Specific recognition of linear polyubiquitin by A20 zinc finger 7 is involved in NF-κB regulation.

LUBAC (linear ubiquitin chain assembly complex) activates the canonical NF-κB pathway through linear polyubiquitination of NEMO (NF-κB essential modulator, also known as IKKγ) and RIP1. However, the regulatory mechanism of LUBAC-mediated NF-κB activation remains elusive. Here, we show that A20 suppresses LUBAC-mediated NF-κB activation by binding linear polyubiquitin via the C-terminal seventh zinc finger (ZF7), whereas CYLD suppresses it through deubiquitinase (DUB) activity. We determined the crystal structures of A20 ZF7 in complex with linear diubiquitin at 1.70-1.98 Å resolutions. The crystal structures revealed that A20 ZF7 simultaneously recognizes the Met1-linked proximal and distal ubiquitins, and that genetic mutations associated with B cell lymphomas map to the ubiquitin-binding sites. Our functional analysis indicated that the binding of A20 ZF7 to linear polyubiquitin contributes to the recruitment of A20 into a TNF receptor (TNFR) signalling complex containing LUBAC and IκB kinase (IKK), which results in NF-κB suppression. These findings provide new insight into the regulation of immune and inflammatory responses.

Long-term clinical results of Charnley total hip arthroplasty using a matte satin-finished stem: a 30-year average follow-up study.

BACKGROUND: Although 50 years have passed since Charnley started performing total hip arthroplasties, there are relatively few patient studies with a follow-up period of over 30 years. We studied radiograms of patients who had undergone Charnley total hip arthroplasty between 1975 and 1981 and evaluated the results at 26-35 years (mean 30.5 years) after the operation. METHODS: The operations were performed in 87 joints of 82 patients (26 males and 56 females) aged 26-81 years (mean age 56 years) at the time of surgery. A monoblock-type Charnley stem with a 22-mm head was used in all patients. The stem had a matte satin surface finish and a flange or round-back shoulder. The socket was made of ultra-high-density polyethylene without a flange. All operations were performed by one surgeon using the original Charnley method in all patients, excluding the first five. RESULTS: Thirty years after the operation, 48 patients have died, 5 had an infection, 9 were lost to follow-up, and 20 patients with 24 joints are still alive. In 77 patients (82 joints), after excluding the 5 with infection, the socket and stem survival rates were 54 and 75% at 30 years after the operation when radiographic loosening or revision surgery was defined as the end point. With revision surgery as the end point, the rates were 68 and 82%, respectively. CONCLUSIONS: These results were as favorable as those with surgical techniques (including the first-generation cement technique) and implants in the early period. Although new techniques and implants have since been introduced, their long-term results should exceed those in this study. The results of this study should provide a valuable index with which to compare the long-term studies of current total hip arthroplasty.

High Photosynthetic Photon Flux Density Differentially Improves Edible Biomass Space Use Efficacy in Edamame and Dwarf Tomato.

Improving edible biomass space use efficacy (EBSUE) is important for sustainably producing edamame and dwarf tomatoes in plant factories with artificial light. Photosynthetic photon flux density (PPFD) may increase EBSUE and space use efficacy (SUE). However, no study has quantitatively explained how PPFD affects EBSUE in edamame and dwarf tomatoes. This study aimed to quantitatively validate the effects of PPFD on EBSUE in dwarf tomatoes and edamame and verify whether this effect differs between these crops. The edamame and dwarf tomato cultivars 'Enrei' and 'Micro-Tom', respectively, were cultivated under treatments with PPFDs of 300, 500, and 700 µmol m-2 s-1. The results showed that the EBSUE and SUE increased with increasing PPFD in both crops. The EBSUE increased depending on the increase in SUE, the dry mass ratio of the edible part to the total plant in the edamame, and the SUE only in the dwarf tomatoes. In conclusion, a high PPFD can improve the EBSUE and SUE of edamame and dwarf tomatoes in different ways at the reproductive growth stage. The findings from this study offer valuable information on optimizing space and resource usage in plant factories with artificial light and vertical farms. Additionally, they shed light on the quantitative impact of PPFD on both EBSUE and SUE.

Effect of red and blue light versus white light on fruit biomass radiation-use efficiency in dwarf tomatoes.

The effect of the ratio of red and blue light on fruit biomass radiation-use efficiency (FBRUE) in dwarf tomatoes has not been well studied. Additionally, whether white light offers a greater advantage in improving radiation-use efficiency (RUE) and FBRUE over red and blue light under LED light remains unknown. In this study, two dwarf tomato cultivars ('Micro-Tom' and 'Rejina') were cultivated in three red-blue light treatments (monochromatic red light, red/blue light ratio = 9, and red/blue light ratio = 3) and a white light treatment at the same photosynthetic photon flux density of 300 µmol m-2 s-1. The results evidently demonstrated that the red and blue light had an effect on FBRUE by affecting RUE rather than the fraction of dry mass partitioned into fruits (Ffruits). The monochromatic red light increased specific leaf area, reflectance, and transmittance of leaves but decreased the absorptance and photosynthetic rate, ultimately resulting in the lowest RUE, which induced the lowest FBRUE among all treatments. A higher proportion of blue light (up to 25%) led to a higher photosynthetic rate, resulting in a higher RUE and FBRUE in the three red-blue light treatments. Compared with red and blue light, white light increased RUE by 0.09-0.38 g mol-1 and FBRUE by 0.14-0.25 g mol-1. Moreover, white light improved the Ffruits in 'Rejina' and Brix of fruits in 'Micro-Tom' and both effects were cultivar-specific. In conclusion, white light may have greater potential than mixed red and blue light for enhancing the dwarf tomato FBRUE during their reproductive growth stage.

MucoRice-CTB line 19A, a new marker-free transgenic rice-based cholera vaccine produced in an LED-based hydroponic system.

We previously established the selection-marker-free rice-based oral cholera vaccine (MucoRice-CTB) line 51A for human use by Agrobacterium-mediated co-transformation and conducted a double-blind, randomized, placebo-controlled phase I trial in Japan and the United States. Although MucoRice-CTB 51A was acceptably safe and well tolerated by healthy Japanese and U.S. subjects and induced CTB-specific antibodies neutralizing cholera toxin secreted by Vibrio cholerae, we were limited to a 6-g cohort in the U.S. trial because of insufficient production of MucoRice-CTB. Since MucoRice-CTB 51A did not grow in sunlight, we re-examined the previously established marker-free lines and selected MucoRice-CTB line 19A. Southern blot analysis of line 19A showed a single copy of the CTB gene. We resequenced the whole genome and detected the transgene in an intergenic region in chromosome 1. After establishing a master seed bank of MucoRice-CTB line 19A, we established a hydroponic production facility with LED lighting to reduce electricity consumption and to increase production capacity for clinical trials. Shotgun MS/MS proteomics analysis of MucoRice-CTB 19A showed low levels of α-amylase/trypsin inhibitor-like proteins (major rice allergens), which was consistent with the data for line 51A. We also demonstrated that MucoRice-CTB 19A had high oral immunogenicity and induced protective immunity against cholera toxin challenge in mice. These results indicate that MucoRice-CTB 19A is a suitable oral cholera vaccine candidate for Phase I and II clinical trials in humans, including a V. cholerae challenge study.

Analysis of nuclear factor-κB (NF-κB) essential modulator (NEMO) binding to linear and lysine-linked ubiquitin chains and its role in the activation of NF-κB.

Nuclear factor-κB (NF-κB) essential modulator (NEMO), a component of the inhibitor of κB kinase (IKK) complex, controls NF-κB signaling by binding to ubiquitin chains. Structural studies of NEMO provided a rationale for the specific binding between the UBAN (ubiquitin binding in ABIN and NEMO) domain of NEMO and linear (Met-1-linked) di-ubiquitin chains. Full-length NEMO can also interact with Lys-11-, Lys-48-, and Lys-63-linked ubiquitin chains of varying length in cells. Here, we show that purified full-length NEMO binds preferentially to linear ubiquitin chains in competition with lysine-linked ubiquitin chains of defined length, including long Lys-63-linked deca-ubiquitins. Linear di-ubiquitins were sufficient to activate both the IKK complex in vitro and to trigger maximal NF-κB activation in cells. In TNFα-stimulated cells, NEMO chimeras engineered to bind exclusively to Lys-63-linked ubiquitin chains mediated partial NF-κB activation compared with cells expressing NEMO that binds to linear ubiquitin chains. We propose that NEMO functions as a high affinity receptor for linear ubiquitin chains and a low affinity receptor for long lysine-linked ubiquitin chains. This phenomenon could explain quantitatively distinct NF-κB activation patterns in response to numerous cell stimuli.

The intertransmembrane region of Kaposi's sarcoma-associated herpesvirus modulator of immune recognition 2 contributes to B7-2 downregulation.

Kaposi's sarcoma-associated herpesvirus (KSHV), a human tumor virus, encodes two homologous membrane-associated E3 ubiquitin ligases, modulator of immune recognition 1 (MIR1) and MIR2, to evade host immunity. Both MIR1 and MIR2 downregulate the surface expression of major histocompatibility complex class I (MHC I) molecules through ubiquitin-mediated endocytosis followed by lysosomal degradation. Since MIR2 additionally downregulates a costimulatory molecule (B7-2) and an integrin ligand (intercellular adhesion molecule 1 [ICAM-1]), MIR2 is thought to be a more important molecule for immune evasion than MIR1; however, the molecular basis of the MIR2 substrate specificity remains unclear. To address this issue, we determined which regions of B7-2 and MIR2 are required for MIR2-mediated B7-2 downregulation. Experiments with chimeras made by swapping domains between human B7-2 and CD8α, a non-MIR2 substrate, and between MIR1 and MIR2 demonstrated a significant contribution of the juxtamembrane (JM) region of B7-2 and the intertransmembrane (ITM) region of MIR2 to MIR2-mediated downregulation. Structure prediction and mutagenesis analyses indicate that Phe119 and Ser120 in the MIR2 ITM region and Asp244 in the B7-2 JM region contribute to the recognition of B7-2 by MIR2. This finding provides new insight into the molecular basis of substrate recognition by MIR family members.

Exploring tomato gene functions based on coexpression modules using graph clustering and differential coexpression approaches.

Gene-to-gene coexpression analysis provides fundamental information and is a promising approach for predicting unknown gene functions in plants. We investigated various associations in the gene expression of tomato (Solanum lycopersicum) to predict unknown gene functions in an unbiased manner. We obtained more than 300 microarrays from publicly available databases and our own hybridizations, and here, we present tomato coexpression networks and coexpression modules. The topological characteristics of the networks were highly heterogenous. We extracted 465 total coexpression modules from the data set by graph clustering, which allows users to divide a graph effectively into a set of clusters. Of these, 88% were assigned systematically by Gene Ontology terms. Our approaches revealed functional modules in the tomato transcriptome data; the predominant functions of coexpression modules were biologically relevant. We also investigated differential coexpression among data sets consisting of leaf, fruit, and root samples to gain further insights into the tomato transcriptome. We now demonstrate that (1) duplicated genes, as well as metabolic genes, exhibit a small but significant number of differential coexpressions, and (2) a reversal of gene coexpression occurred in two metabolic pathways involved in lycopene and flavonoid biosynthesis. Independent experimental verification of the findings for six selected genes was done using quantitative real-time polymerase chain reaction. Our findings suggest that differential coexpression may assist in the investigation of key regulatory steps in metabolic pathways. The approaches and results reported here will be useful to prioritize candidate genes for further functional genomics studies of tomato metabolism.

The Pavlik harness in the treatment of developmentally dislocated hips: results of Japanese multicenter studies in 1994 and 2008.

BACKGROUND: It has already been more than 50 years since the Pavlik harness was introduced in Japan, and today the Pavlik harness is widely recognized as the standard initial treatment modality for developmental dysplasia of the hip. We performed a multicenter nationwide questionnaire study concerning the results of Pavlik harness treatment twice in 1994 and 2008. METHODS: In 1994 and in 2008, we sent questionnaires to 12 institutes in Japan specializing mainly in pediatric orthopedics. We compare the results of these two studies and discuss differences in reduction rates, incidence of avascular necrosis in the femoral epiphysis and the percentage of joints with acceptable morphology (Severin grade I + II/total) at skeletal maturity. We statistically assessed these results to see whether there were changes in the treatment outcomes over this 14-year period. RESULTS: Reduction of the dislocated hips was obtained by the Pavlik harness in 80.2% (1990/2481 hips; 1994) and 81.9% (1248/1523 hips; 2008). The incidences of avascular necrosis of the proximal femoral epiphysis in the dysplastic hips were 14.3% (119/835 hips; 1994) and 11.5% (76/663 hips; 2008). The type of avascular necrosis in hips from the 2008 study was determined according to the classification of Kalamchi and MacEwen: 24/69 hips (34.8%) were classified as group I; 20/69 hips (29.0%) as group II; 11/69 hips (15.9%) as group Ill; 14/69 hips (20.3%) as group IV. The percentages of hips with acceptable outcomes at skeletal maturity discerned from Severin X-ray changes (grade I + II/total) were 72.3% (604/835 hips; 1994) and 77.7% (488/628 hips; 2008). CONCLUSION: Reduction rates and the incidence of avascular necrosis in 2008 were statistically similar to the results in 1994. The rate of acceptable outcome (Severin grade I + II/total) in 2008 was statistically higher than that of 1994.

Evaluation of the enhancement of photosynthetic rate in a komatsuna (Brassica rapa L. var. perviridis) canopy with upward lighting using an optical simulation in a plant factory with artificial light.

In a plant factory with artificial light (PFAL), upward lighting is expected to prevent senescence and decrease in the photosynthetic capacity of the lower leaves in the canopy. Upward lighting may also increase the photosynthetic rate of a canopy by improving its photosynthetic photon flux density (PPFD) distribution. However, the net photosynthetic rate (Pn) of leaves is lower when the abaxial surface is irradiated than that when the adaxial surface is irradiated. The aim of this study was to estimate the PPFD in a PFAL and the Pn of plants using three-dimensional plant models and optical simulation. First, we measured the Pn of komatsuna (Brassica rapa L. var. perviridis) leaves under different conditions of the proportion (pad ) of PPFD on the adaxial surface to total PPFD on both surfaces and developed an equation for the light response curve of photosynthesis considering pad . When PPFD was low, except when it was 30 and 70 µmol m-2 s-1, Pn increased as pad increased, because the absorptance also increased with pad . Under high PPFD conditions, Pn was maximized at 67-83% of pad because the light would be distributed more efficiently for photosynthesis. Next, using optical simulation and the developed equation, we estimated the photosynthetic rate of a komatsuna canopy (CPn) under downward and upward lighting. The CPn increased by 1.08-1.13 times by combining downward and upward lighting due to the increase in the photosynthetic photon flux (PPF) of light incident on the canopy and the decrease in the spatial variation of PPFD on the leaves in the canopy. As the depreciation of lamps for upward lighting accounts for 7.5-9.0% of the production cost in a PFAL, even if the depreciation of lamps for upward lighting increased, enhancement of CPn by upward lighting would be cost-effective. We performed optical simulations under 220 conditions and evaluated them using CPn as an index. Moreover, we provided the proportion of PPF of upward lighting that improved CPn and discussed the reason for this improvement. The result shows that optical simulation is useful for evaluating the lighting design in a PFAL and analyzing the effects of the lighting design on the light environment and photosynthesis.

Photosynthetic photon flux density affects fruit biomass radiation-use efficiency of dwarf tomatoes under LED light at the reproductive growth stage.

This study aimed to analyze the effects of photosynthetic photon flux density (PPFD) on fruit biomass radiation-use efficiency (FBRUE) of the dwarf tomato cultivar 'Micro-Tom' and to determine the suitable PPFD for enhancing the FBRUE under LED light at the reproductive growth stage. We performed four PPFD treatments under white LED light: 200, 300, 500, and 700 µmol m-2 s-1. The results demonstrated that a higher PPFD led to higher fresh and dry weights of the plants and lowered specific leaf areas. FBRUE and radiation-use efficiency (RUE) were the highest under 300 µmol m-2 s-1. FBRUE decreased by 37.7% because RUE decreased by 25% and the fraction of dry mass portioned to fruits decreased by 16.9% when PPFD increased from 300 to 700 µmol m-2 s-1. Higher PPFD (500 and 700 µmol m-2 s-1) led to lower RUE owing to lower light absorptance, photosynthetic quantum yield, and photosynthetic capacity of the leaves. High source strength and low fruit sink strength at the late reproductive growth stage led to a low fraction of dry mass portioned to fruits. In conclusion, 300 µmol m-2 s-1 PPFD is recommended for 'Micro-Tom' cultivation to improve the FBRUE at the reproductive growth stage.

The new Japanese postgraduate medical education and quality of emergency medical care.

BACKGROUND: The new postgraduate medical education (PGME) was recently introduced to improve quality of emergency care in Japan. OBJECTIVES: To compare the quality of care and confidence in provision of emergency medicine between physicians who completed the old and new PGME programs. METHODS: A cross-sectional survey was sent to 279 physicians of postgraduate years 4-9, and 208 responses (75%) were received. Quality of care in emergency medicine was measured using 26 questions on treatment choices for various clinical conditions. Each question had six responses, including a single correct choice. Effect size was obtained by dividing the total difference in score by the standard deviation of the score distribution. Confidence in emergency medicine was rated using four self-reported items on the level of confidence in treating acute illnesses in various emergency medicine settings. RESULTS: The mean score for quality of care was significantly higher in the new PGME group (15.3) compared to the old PGME group (12.8). The difference in scores was 2.5 (p < 0.01) and the effect size (0.47) indicated a moderate difference. Linear regression of total scores adjusted for physician covariates produced similar results of an adjusted score difference of 2.5 (p < 0.01) and an adjusted effect size of 0.47. The new PGME group also had significantly greater confidence in provision of emergency medicine based on significant differences between the groups for all four self-reported items (all p < 0.05). CONCLUSIONS: Japanese physicians who complete the new PGME program are likely to provide higher quality of care and have greater confidence in emergency medicine compared to those who completed the old PGME program.

Ozone control as a novel method to improve health-promoting bioactive compounds in red leaf lettuce (Lactuca sativa L.).

In this study, we determined the short-term effects of ozone exposure on the growth and accumulation of bioactive compounds in red lettuce leaves grown in a controlled environment plant factory with artificial light, also known as a vertical farm. During cultivation, twenty-day-old lettuce (Lactuca sativa L. var. Redfire) seedlings were exposed to 100 and 200 ppb of ozone concentrations for 72 h. To find out how plants react to ozone and light, complex treatments were done with light and ozone concentrations (100 ppb; 16 h and 200 ppb; 24 h). Ozone treatment with 100 ppb did not show any significant difference in shoot fresh weight compared to that of the control, but the plants exposed to the 200 ppb treatment showed a significant reduction in fresh weight by 1.3 fold compared to the control. The expression of most genes in lettuce plants exposed to 100 and 200 ppb of ozone increased rapidly after 0.5 h and showed a decreasing trend after reaching a peak. Even when exposed to a uniform ozone concentration, the pattern of accumulating bioactive compounds such as total phenolics, antioxidant capacity and total flavonoids varied based on leaf age. At a concentration of 200 ppb, a greater accumulation was found in the third (older) leaf than in the fourth leaf (younger). The anthocyanin of lettuce plants subjected to 100 and 200 ppb concentrations increased continuously for 48 h. Our results suggest that ozone control is a novel method that can effectively increase the accumulation of bioactive compounds in lettuce in a plant factory.