Classification of Southeast Asian mints (Mentha spp.) based on simple sequence repeat markers.

Mentha is a complex genus encompassing many species as a consequence of their interspecific hybridization and polyploidy. Southeast Asian mints have been poorly distinguished though they are widely used for culinary and medical purposes. In this study, we have analyzed Southeast Asian mints and known varieties as well as a related Lamiaceae species (Nepeta sp.) using simple sequence repeat (SSR) markers and leaf morphology. Two types of mints were clearly distinguished based on their venation pattern and leaf shape index. We developed 12 SSR markers that allowed good amplification in the Mentha and another Lamiaceae species. In the SSR-based phylogram, the Mentha lines could be delimited into groups I-VI. The Southeast Asian mints divided into groups I and II, and the phylogram separated most of the available species, with groups I and II containing the known species M. × cordifolia and M. arvensis, respectively. The separation of the two groups was supported by a population structure analysis. The SSR markers developed in this study enabled the simultaneous classification of mints and will help improve our understanding of the genetic composition of known mint varieties and as yet unclassified Southeast Asian mints.

Endocrine mucin-producing sweat gland carcinoma of the cheek.

Endocrine mucin-producing sweat gland carcinoma (EMPSGC) is a low-grade adnexal malignant neoplasm. We report a 90-year-old man who had a hard, dome-shaped tumor approximately 9 mm in diameter on the left cheek. Dermoscopy showed an overall, non-uniformly light-pink tumor with crust. The diagnosis of EMPSGC is made histologically from excisional biopsy. No signs of recurrent disease were evident at 42 months postoperatively.

Toxic effects of 4-methylthio-3-butenyl isothiocyanate (Raphasatin) in the rat urinary bladder without genotoxicity.

We recently reported that 4-methylthio-3-butenyl isothiocyanate (MTBITC) exerts chemopreventive effects on the rat esophageal carcinogenesis model at a low dose of 80 ppm in a diet. In contrast, some isothiocyanates (ITCs) have been reported to cause toxic effects, promotion activity, and/or carcinogenic potential in the urinary bladder of rats. In the present study, we investigated whether MTBITC had toxic effects in the urinary bladder similar to other ITCs, such as phenethyl ITC (PEITC). First, to examine the early toxicity of MTBITC, rats were fed a diet supplemented with 100, 300 or 1000 ppm MTBITC for 14 days. Treatment with 1000 ppm MTBITC caused increased organ weights and histopathological changes in the urinary bladder, producing lesions similar to those of 1000 ppm PEITC. In contrast, rats treated with 100 or 300 ppm MTBITC showed no signs of toxicity. Additionally, we performed in vivo genotoxicity studies to clarify whether MTBITC may exhibit a carcinogenic potential through a genotoxic mechanism in rats. Rats were treated with MTBITC for 3 days at doses of 10, 30 or 90 mg kg-1 body weight by gavage, and comet assays in the urinary bladder and micronucleus assays in the bone marrow were performed. No genotoxic changes were observed after treatment with MTBITC at all doses. Overall, these results suggested that the effects of MTBITC in the rat urinary bladder are less than those of PEITC, but that MTBITC could have toxic effects through a nongenotoxic mechanism in the urinary bladder of rats at high doses. Copyright © 2016 John Wiley & Sons, Ltd.

4-Methylthio-3-butenyl isothiocyanate (raphasatin) exerts chemopreventive effects against esophageal carcinogenesis in rats.

To examine the effects of 4-methylthio-3-butenyl isothiocyanate on esophageal carcinogenesis, male 6-week-old F344 rats were subcutaneously injected with 0.5 mg/kg body weight N-nitrosomethylbenzylamine three times per week for 5 weeks and fed a diet supplemented with 80 ppm 4-methylthio-3-butenyl isothiocyanate, equivalent to 6.05 mg/kg body weight/day for the initiation stage, 4.03 mg/kg body weight/day for the promotion stage, or 4.79 mg/kg body weight/day for all stages. Although the incidence of lesions was not affected by 4-methylthio-3-butenyl isothiocyanate treatment, the multiplicity of squamous cell papilloma in the esophagus was significantly decreased in rats in the 4-methylthio-3-butenyl isothiocyanate initiation stage group (1.13 ± 0.74), 4-methylthio-3-butenyl isothiocyanate promotion stage group (1.47 ± 0.99), and 4-methylthio-3-butenyl isothiocyanate all stage group (1.47 ± 1.13) as compared with rats treated with N-nitrosomethylbenzylamine alone (3.00 ± 1.46). Immunohistochemical analysis revealed that 4-methylthio-3-butenyl isothiocyanate induced apoptosis, suppressed cell proliferation, and increased p21 expression when administered in the promotion phase. These modifying effects were not observed in the rats treated with 4-methylthio-3-butenyl isothiocyanate alone. Our results indicated that 4-methylthio-3-butenyl isothiocyanate may exert chemopreventive effects against N-nitrosomethylbenzylamine-induced esophageal carcinogenesis in rats.

Factors associated with changes in the objectively measured physical activity among Japanese adults: A longitudinal and dynamic panel data analysis.

Factors associated with dynamic changes in the objectively measured physical activity have not been well understood. We aimed to 1) evaluate the longitudinal change in the physical activity trajectory according to sex which is associated with age and to 2) determine the factors associated with the dynamic change in physical activity-related variables across a wide age range among Japanese adults. This longitudinal prospective study included 689 Japanese adults (3914 measurements) aged 26-85 years, whose physical activity data in at least two surveys were available. Physical activity-related variables, such as intensity (inactive, light [LPA; 1.5 to 2.9 metabolic equivalents (METs)], moderate-to-vigorous [MVPA; ≥3.0 METs]), total energy expenditure (TEE), physical activity level (PAL), and step count, were evaluated using a validated triaxial accelerometer. Statistical analysis involved the latent growth curve models and random-effect panel data multivariate regression analysis. During a mean follow-up period of 6.8 years, physical activity was assessed an average of 5.1 times in men and 5.9 times in women. The profiles for the inactive time, LPA (only men), MVPA, step count, PAL, and TEE showed clear curvature, indicating an accelerated rate of change around the age of 70. In contrast, other variables exhibited minimal or no curvature over the age span. The MVPA trajectory was positively associated with alcohol consumption, hand grips, leg power, and trunk flexibility and negatively associated with age, local area, body mass index (BMI), comorbidity score, and heart rate over time. Our results indicated that the physical activity trajectory revealed clear curvature, accelerated rate of change around the age of 70, and determined physical health and fitness and BMI as dynamic factors associated with physical activity changes. These findings may be useful to help support populations to achieve and maintain the recommended level of physical activity.

Methylthioacetic acid, a derivative of aroma compounds from Cucumis melo var. conomon dose-dependently triggers differentiation and apoptosis of RCM-1 human colorectal cancer cells.

Methylthioacetic acid (MTA) is an acid-hydrolyzed derivative of a natural aroma compound, methylthioacetic acid ethyl ester isolated from Cucumis melo var. conomon (Katsura-uri, Japanese Picking Melon), and induces a villiform-like structure dome in RCM-1 human colorectal cancer cell culture. Thus far, the physiological and molecular properties of MTA-mediated dome formation remain unknown. Herein, MTA (not more than 2 mM) was demonstrated to differentiate the unorganized cell mass into the dome in RCM-1 cell culture by disclosing the correlation between dome formation and several intestinal differentiation markers such as alkaline phosphatase activity and the protein levels of dipeptidyl peptidase 4, villin, and Krüppel-like factor 4. Dome formation in RCM-1 cell culture was additively enhanced by the simultaneous administration of MTA and butyric acid (BA), suggesting that MTA directs the differentiation of RCM-1 cells, potentially through the same or similar pathway(s) shared with BA. Notably, a high dose of MTA (2 mM or more) elevated several apoptosis markers, such as DNA fragmentation, caspase-3/7 activity, and cleavage of poly(ADP-ribose) polymerase. Altogether, in addition to RCM-1 cell differentiation, MTA triggers apoptosis. These results indicate that MTA is a potential anticarcinogenic agent applicable in differentiation therapy and traditional chemotherapy against colorectal cancers.

Behavior and possible function of Arabidopsis BES1/BZR1 homolog 2 in brassinosteroid signaling.

Two key transcription factors (TFs) in brassinosteroid (BR) signaling BRASSINOSTEROID INSENSITIVE 1-EMS-SUPPRESSOR 1 (BES1) and BRASSINAZOLE RESISTANT 1 (BZR1), belong to a small family with four BES1/BZR1 homologs (BEH1-4). To date, in contrast to the wealth of knowledge regarding BES1 and BZR1, little is known about BEH1-4. Here, we show that BEH2 was expressed preferentially in the roots and leaf margins including serrations, which was quite different from another member BEH4, and that BRs downregulated BEH2 through a module containing GSK3-like kinases and BES1/BZR1 TFs, among which BES1, rather than BZR1, contributed to this process. In addition, BEH2 consistently existed in the nucleus, suggesting that its subcellular localization is not under BR-dependent nuclear-cytoplasmic shuttling control. Furthermore, gene ontology analysis on RNA-seq data indicated that BEH2 may be implicated in stress response and photosynthesis. These findings might assist in the future elucidation of the molecular mechanisms underlying BR signaling.

Association Between Temporal Changes in Diet Quality and Concurrent Changes in Dietary Intake, Body Mass Index, and Physical Activity Among Japanese Adults: A Longitudinal Study.

BACKGROUND: Many cross-sectional studies have identified modifiable factors such as dietary intake, physique, and physical activity associated with diet quality but were unable to determine how a specific individual's diet quality changes with these factors. These relationships may vary depending on an individual's dietary intake. We aimed to determine the association between temporal changes in diet quality and concurrent changes in dietary intake, body mass index (BMI), and physical activity according to the diet quality trajectory pattern. METHODS: This longitudinal prospective study included 697 Japanese adults aged 26-85 years, at baseline, with available data from at least two dietary intake surveys (4,118 measurements). Dietary intake and physical activity were evaluated using validated dietary questionnaires and a triaxial accelerometer. Diet quality was calculated using the Nutrient-Rich Food Index 9.3 (NRF9.3), while physical activity was calculated based on the duration of activity performed at each level of intensity (sedentary, light, moderate, and vigorous). Body mass index was calculated from the measured height and weight. Statistical analyses involved latent class growth models (LCGM) and random-effect panel data analysis. RESULTS: During a mean follow-up period of 6.8 years, NRF9.3 scores were assessed, on average, 5.4 times in men and 6.1 times in women. Based on the NRF9.3 score, three separate trajectory groups-"low-increasing," "medium-increasing," and "high-stable"-among individuals aged 26-90 years were identified using LCGM. In the multivariate analysis, the NRF9.3 score trajectory was positively associated with intake of energy, protein, dietary fiber, vitamins A and C, magnesium, and food items, such as fruits and vegetables, and was negatively associated with BMI and the intake of added sugar, saturated fats, sodium, and food items, such as meat and sugar and confectioneries, even after adjusting for covariates. These relationships displayed heterogeneity across the identified NRF9.3 score trajectory groups. In the low-increasing group, an inverse relationship was observed between sedentary behavior and NRF9.3 score trajectory. CONCLUSIONS: We identified modifiable factors associated with temporal changes in diet quality across a wide age range; however, these factors may vary according to the diet quality trajectories. Our findings may help develop effective strategies for improving diet quality, according to the trajectory of diet quality.

Gray-edged line sign of scabies burrow.

A scabies burrow is created by a mature female mite laying eggs through the stratum corneum, representing a kind of scabies eruption. We have noticed that the edges of the scabies burrow sometime appear as blackish-gray lines. We named these lines the "gray-edged line" sign, as a new feature of scabies burrows. The gray-edged line sign has the following two tendencies: (i) it is rarely seen on the palm or sole; and (ii) when the burrow follows a curved course, the gray-edged line often forms on the outer wall. Explaining the formation of this sign from clinical findings was difficult, so the aim of the present study was to elucidate the mechanisms underlying the gray-edged line sign. This retrospective study involved collection of data from electronic medical records of patients treated for scabies in our department between April 2015 and February 2020. We treated 32 scabies patients, including 4 patients with the gray-edged line sign. We analyzed clinical features, dermoscopy, histopathology and special stains. Fontana-Masson staining showed melanin staining in three parts: feces; some keratinocytes around the scabies burrows; and the mouth and legs of the scabies mite. The gray-edged line sign appears to represent mite feces containing melanin.

Expression profiles of four BES1/BZR1 homologous genes encoding bHLH transcription factors in Arabidopsis.

Arabidopsis bHLH-type transcription factors-BRASSINOSTEROID INSENSITIVE 1-EMS-SUPPRESSOR 1 (BES1) and BRASSINAZOLE RESISTANT 1 (BZR1)-play key roles in brassinosteroid (BR) signaling. By contrast, the functions of the other four BES1/BZR1 homologs (BEH1-4) remain unknown. Here, we describe the detailed expression profiles of the BES1/BZR1 family genes. Their expressions were distinct regarding growth-stage dependence and organ specificity but exhibited some overlaps as well. Furthermore, their mRNA levels mostly remained unchanged responding to seven non-BR phytohormones. However, BEH1 and BEH2 were downregulated by brassinolide, suggesting a close association with the BR function. Additionally, BEH4 was ubiquitously expressed throughout the life of the plant but displayed some expression preference. For instance, BEH4 expression was limited to guard cells and the adjacent pavement cells in the leaf epidermis and was induced during growth progression in very young seedlings, suggesting that BEH4 is specifically regulated in certain contexts, although it is almost constitutively controlled.

Chemical and molecular bases of dome formation in human colorectal cancer cells mediated by sulphur compounds from Cucumis melo var. conomon.

Colorectal cancer was the third most commonly diagnosed malignant tumor and the fourth leading cause of cancer deaths worldwide in 2012. A human colorectal cancer cell line, RCM-1, was established from a colon cancer tissue diagnosed as a well-differentiated rectum adenocarcinoma. RCM-1 cells spontaneously form 'domes' (formerly designated 'ducts') resembling villiform structures. Two sulphur-containing compounds from Cucumis melo var. conomon (Katsura-uri, or Japanese pickling melon), referred to as 3-methylthiopropionic acid ethyl ester (MTPE) and methylthioacetic acid ethyl ester (MTAE), can induce the differentiation of the unorganized cell mass of an RCM-1 human colorectal cancer cell culture into a dome. However, the underlying molecular mechanisms of such dome formation have not been previously reported. Here, we performed a structure-activity relationship analysis, which indicated that methylthioacetic acid (MTA) was the lowest molecular weight compound with the most potent dome-inducing activity among 37 MTPE and MTAE analogues, and the methylthio group was essential for this activity. According to our microarray analysis, MTA resulted in down-regulation of 537 genes and up-regulation of 117 genes. Furthermore, MTA caused down-regulation of many genes involved in cell-cycle control, with the cyclin E2 (CCNE2) and cell division cycle 25A (CDC25A) genes being the most significantly reduced. Pharmacological analysis showed that the administration of two cell-cycle inhibitors for inactivating CDC25A phosphatase (NSC95397) and the cyclin E2/cyclin-dependent kinase 2 complex (purvalanol A) increased the dome number independently of MTA. Altogether, our results indicate that MTA is the minimum unit required to induce dome formation, with the down-regulation of CDC25A and possibly CCNE2 being important steps in this process.

Contribution of Katsura-uri (Japan's Heirloom Pickling Melon, Cucumis melo var. conomon) at the Completely Ripe Stage to Diabetes Control.

The fruit of Katsura-uri (Japan's heirloom pickling melon, Cucumis melo var. conomon) possesses a fruity aroma and moderate sweetness. The fruit juice has potential to minimize human postprandial blood glucose levels. This study provides information regarding the health benefits of Katsura-uri and its utility in treating diabetes. The study methodology involved measuring the color and firmness of Katsura-uri fruit at five ripening stages, and quantitation of the aroma substances, proximate composition, and sugars. Significant changes were detected in the color, firmness, and level of aroma substances with ripening of Katsura-uri fruit, albeit with no major changes in proximate composition, with the exception of dietary fiber, and sugars. To determine the effects of Katsura-uri juice, the blood glucose levels of ten diabetic volunteers aged 46-75 y were monitored after its consumption, and compared with after consumption of muskmelon juice equivalent to the total weight of Katsura-uri juice. The blood glucose area under the curve level was significantly lower after consumption of Katsura-uri juice (16±5 h • mg/dL) than after consumption of muskmelon juice (55±17 h • mg/dL; p<0.05). The level of the glucose spike was also significantly lower after consumption of Katsura-uri juice (22±5 mg/dL) than after consumption of muskmelon juice (64±6 mg/dL; p<0.05). The completely ripe Katsura-uri fruit provides the best results for diabetic subjects, which is the first case of fruits sweetened with the addition of zero-calorie sweeteners.

Investigation on the Epoxidation of Piperitenone, and Structure-activity Relationships of Piperitenone Oxide for Differentiation-inducing Activity.

Piperitenone oxide, a major chemical constituent of the essential oil of spearmint, Mentha spicata, induces differentiation in human colon cancer RCM-1 cells. In this study, piperitenone oxide and trans-piperitenone dioxide were prepared as racemic forms by epoxidation of piperitenone. The relative configuration between two epoxides in piperitenone dioxide was determined to be trans by 1H NMR analysis and nuclear Overhauser effect spectroscopy (NOESY) in conjunction with density functional theory (DFT) calculations. Optical resolution of (±)-piperitenone oxide by high-performance liquid chromatography (HPLC) using a chiral stationary phase (CSP) afforded both enantiomers with over 98% enantiomeric excess (ee). Evaluation of the differentiation-inducing activity of the synthetic compounds revealed that the epoxide at C-1 and C-6 in piperitenone oxide is important for the activity, and (+)-piperitenone oxide has stronger activity than (-)-piperitenone oxide. The results obtained in this study provide new information on the application of piperitenone oxide and spearmint for differentiation-inducing therapy. Furthermore, natural piperitenone oxide was isolated from M. spicata. The enantiomeric excess of the isolated natural piperitenone oxide was 66% ee. Epoxidation of piperitenone with hydrogen peroxide proceeded in a phosphate buffer under weak basic conditions to give (±)-piperitenone oxide. These results suggest that the nonenzymatic epoxidation of piperitenone, which causes a decrease in the enantiomeric excess of natural piperitenone oxide, is accompanied by an enzymatic epoxidation in the biosynthesis of piperitenone oxide.

Erratum to: The effect of Katsura-uri (Japanese pickling melon, Cucumis melo var. conomon) and its derived ingredient methylthioacetic acid on energy metabolism during aerobic exercise.

[This corrects the article DOI: 10.1186/s40064-015-1144-y.].

The effect of Katsura-uri (Japanese pickling melon, Cucumis melo var. conomon) and its derived ingredient methylthioacetic acid on energy metabolism during aerobic exercise.

PURPOSE: We investigated the effect of Katsura-uri (Japanese pickling melon; Cucumis melo var. conomon) on energy metabolism during exercise in human and animal studies. METHODS: Eight healthy men (mean age, 21.4 ± 0.7 years) participated in a single-blind, crossover study. Thirty minutes after ingesting the Katsura-uri drink or placebo drink, they exercised on a cycle ergometer at 40% maximal heart rate for 30 min. Respiratory gas analysis was performed during exercise to examine oxygen consumption and substrate utilization. Blood biochemical parameters were evaluated during exercise. In the animal study, the effect of methylthioacetic acid (MTA), a Katsura-uri derived component was examined in mice. Immediately after running at 25 m/min for 30 min, biochemical parameters in the hind limb muscle and blood of mice were measured. RESULTS: Oxygen consumption during exercise was higher in the Katsura-uri condition (19.8 ± 3.5 mL/kg/min) than the placebo condition (18.6 ± 3.0 mL/kg/min) (P < 0.05). The elevation of blood lactate was lower in the Katsura-uri condition (1.7 ± 0.4 mM) than the placebo condition (2.2 ± 0.6 mM) 15 min after beginning exercise (P < 0.05). There was a higher positive correlation between lactate concentration and carbohydrate oxidation during exercise in the Katsura-uri condition (R(2) = 0.86) compared to the placebo condition (R(2) = 0.47). The decrease in intermuscular pH and the increase in blood lactate following exercise were prevented by MTA supplementation (250 ppm) with significant differences in the MTA-supplemented group compared to the control group. CONCLUSIONS: These results suggest that the ingestion of Katsura-uri and/or MTA improves glucose metabolism and acidification in skeletal muscles during exercise in human and animal studies.

Home-based active video games to promote weight loss during the postpartum period.

PURPOSE: Weight retention during the postpartum period is critical for the later development of obesity in women. Traditional physical activity is frequently discontinued because of incompatibility with mothers' agenda (i.e., baby care). In the present study, active video games (AVG) are proposed for postpartum women to improve their body composition. METHODS: Thirty-four postpartum women (body mass index = 24.5 ± 3.4 kg·m) were randomized to an AVG group or a control group. Subjects assigned to the AVG group were given a Wii Nintendo console with the game Wii Fit Plus for 40 d. The two groups were tested for weight, body mass index, body fat mass, waist and hip circumferences, and other anthropometric parameters. Physical fitness, energy expenditure, energy intake, and adverse events were also investigated. RESULTS: The AVG group lost more weight than the control group (-2.2 ± 0.9 vs. -0.5 ± 0.7 kg, P < 0.001). They also exhibited more important reductions of BMI, waist and hip circumferences, and body fat (P < 0.05). During the 40-d period, subjects expended an estimated 4682 ± 2874 kcal just by playing AVG. Daily energy intake was reduced by 206 ± 559 kcal. There were significant positive correlations between playing frequency, total playing time, total energy expenditure during the 40-d period, and decrease in daily energy intake respectively, and weight loss (P < 0.05). Playing time data suggested no conflict with baby care activities. CONCLUSIONS: AVGs could represent an interesting spare physical activity for postpartum women. In the present study, these games promoted physical activity, induced a reduction of energy intake, and subsequently minimized weight retention.

Association between dietary intake of micronutrients and cardiorespiratory fitness in Japanese men.

Previous studies have demonstrated that meeting the dietary recommendations for macronutrients was significantly associated with higher cardiorespiratory fitness (CRF) levels in adults. However, the relation between the status of micronutrient intake and CRF still remains unclear. This study examined the association between micronutrient intake status (based on adherence to the dietary reference intakes (DRI)) and CRF in Japanese men. The study comprised 373 Japanese men aged 30-69 years. Dietary intake was assessed with a self-administered diet history questionnaire. Overall micronutrient intake status was quantified using an overall nutrient adequacy score (ONAS) for thirteen selected micronutrients. ONAS was calculated based on adherence to the DRI for Japanese. CRF was defined as V̇O2max during a maximal incremental test on a bicycle ergometer. Physical activity was measured using accelerometer-based activity monitors for seven consecutive days. We observed a significant inverse trend for the prevalence of inadequacy for the intake of vitamin A and Ca across incremental CRF categories (P < 0·05). In a multivariate model, the ONAS was positively associated with absolute (ß = 0·10, P = 0·02) and relative V̇O2max (ß = 0·09, P = 0·04), independent of physical activity. The OR for being unfit (the lowest 25 % of the age-specific distribution of V̇O2max) in the third ONAS tertile compared with the first ONAS tertile was 0·52 (95 % CI 0·28, 0·96). These results demonstrated that the intake of several individual micronutrients and overall micronutrient intake status are independently and positively associated with CRF in Japanese men.