O- and N-Methyltransferases in benzylisoquinoline alkaloid producing plants.

BACKGROUND: Secondary metabolites such as benzylisoquinoline alkaloids (BIA) have attracted considerable attention because of their pharmacological properties and potential therapeutic applications. Methyltransferases (MTs) can add methyl groups to alkaloid molecules, altering their physicochemical properties and bioactivity, stability, solubility, and recognition by other cellular components. Five types of O-methyltransferases and two types of N-methyltransferases are involved in BIA biosynthesis. OBJECTIVE: Since MTs may be the source for the discovery and development of novel biomedical, agricultural, and industrial compounds, we performed extensive molecular and phylogenetic analyses of O- and N-methyltransferases in BIA-producing plants. METHODS: MTs involved in BIA biosynthesis were isolated from transcriptomes of Berberis koreana and Caulophyllum robustum. We also mined the methyltransferases of Coptis japonica, Papaver somniferum, and Nelumbo nucifera from the National Center for Biotechnology Information protein database. Then, we analyzed the functional motifs and phylogenetic analysis. RESULT: We mined 42 O-methyltransferases and 8 N-methyltransferases from the five BIA-producing plants. Functional motifs for S-adenosyl-L-methionine-dependent methyltransferases were retained in most methyltransferases, except for the three O-methyltransferases from N. nucifera. Phylogenetic analysis revealed that the methyltransferases were grouped into four clades, I, II, III and IV. The clustering patterns in the phylogenetic analysis suggested a monophyletic origin of methyltransferases and gene duplication within species. The coexistence of different O-methyltransferases in the deep branch subclade might support some cases of substrate promiscuity. CONCLUSIONS: Methyltransferases may be a source for the discovery and development of novel biomedical, agricultural, and industrial compounds. Our results contribute to further understanding of their structure and reaction mechanisms, which will require future functional studies.

Postharvest Quality Improvement of Bell Pepper (Capsicum annuum L. cv Nagano) with Forced-Air Precooling and Modified Atmosphere Packaging.

Optimum postharvest storage conditions increase the postharvest quality and shelf life of horticultural crops. The effects of forced-air precooling (FAP) and modified atmosphere packaging (MAP) on shelf life, physicochemical quality, and health-promoting properties of bell pepper (Capsicum annuum L. cv. Nagano) harvested at 90 and 50% coloring stages in May and July respectively, stored at 11 °C, 95% relative humidity were assessed. Fruits were subjected to four treatments: FAP + 30 µm polyethylene liner (FOLO); FAP-only (FOLX); 30 µm polyethylene liner-only (FXLO); and control (FXLX). The quality attributes, viz. weight loss, firmness, color, soluble solids content (SSC), soluble sugars, total phenolic content (TPC), total flavonoid content (TFC), 2,2-dephenyl-1-picrylhydrazyl (DPPH), and 2,2'-azino-bis-3-ethylbenzo-thiazoline-6-sulfonic acid (ABTS) were evaluated. The investigated parameters differed significantly (p < 0.05) among treatments except for soluble sugars. FOLO maintained sensory quality (weight loss, firmness, and color), physicochemical (SSC and soluble sugars), and health-promoting properties compared to other treatments during storage. The 50% coloring fruits had a huge variation between treatments than 90% coloring. The results revealed more TPC and antioxidant capacity in the 50% than in the 90% coloring fruits. The study highlights the need to consider the ideal fruit coloring stage at harvest under the effect of FAP and MAP treatments in preserving bell pepper's postharvest quality and shelf life.

Scanning accuracy of an intraoral scanner according to different inlay preparation designs.

BACKGROUND: The accuracy of intraoral scanning plays a crucial role in the workflow of computer-assisted design/computer-assisted manufacturing. However, data regarding scanning accuracy for inlay preparation designs are lacking. The purpose of this in vitro study was to evaluate the influence of the depth of the occlusal cavity and width of the gingival floor of the proximal box on the trueness and precision of intraoral scans for inlay restoration. METHODS: Artificial teeth were used in this study. Four types of preparations for mesio-occlusal inlay were performed on each #36 artificial tooth depending on two different depths of the occlusal cavity (1 mm and 2 mm) and widths of the gingival floor of the proximal box (1.5 mm and 2.5 mm). Artificial teeth were scanned 10 times each with Cerec Primescan AC, and another scan was performed subsequently with a laboratory scanner as a reference (n = 10). Standard tessellation language files were analyzed using a three-dimensional analysis software program. Experimental data were analyzed using two-way analysis of variance and the Bonferroni multiple comparison test. RESULTS: The narrow shallow group had significantly higher deviation values for trueness than the wide deep group (p < 0.05). The wide deep group had the lowest average deviation value for trueness and there was no significant difference between the narrow deep and wide shallow groups (p > 0.05). For the mean maximum positive deviation, the wide groups had significantly lower values than the narrow groups (p < 0.05). Trueness was affected by both the width and depth(p < 0.05), whereas the mean maximum positive deviation was affected by the width (p < 0.05). The mean maximum negative deviation was affected by all three factors (p < 0.05). Precision was affected by the depth and the interaction between the depth of the occlusal cavity and width of the gingival floor (p < 0.05). CONCLUSIONS: The design of different inlay cavity configurations affected the accuracy of the digital intraoral scanner. The highest average deviation for trueness was observed in the narrow shallow group and the lowest in the wide deep group. With regard to precision, the narrow shallow group showed the lowest average deviation, and the narrow deep group showed highest value.

Isolation and Characterization of the Genes Involved in the Berberine Synthesis Pathway in Asian Blue Cohosh, Caulophyllum robustum.

Caulophyllum robustum, commonly named Asian blue cohosh, is a perennial herb in the family Berberidaceae. It has traditionally been used for folk medicine in China. We isolated berberine from the leaves, stem, roots, and fruits of C. robustum, and this is the first report on berberine in this species. Transcriptome analysis was conducted for the characterization of berberine biosynthesis genes in C. robustum, in which, all the genes for berberine biosynthesis were identified. From 40,094 transcripts, using gene ontology (GO) analysis, 26,750 transcripts were assigned their functions in the categories of biological process, molecular function, and cellular component. In the analysis of genes expressed in different tissues, the numbers of genes in the categories of intrinsic component of membrane and transferase activity were up-regulated in leaves versus stem. The berberine synthesis genes in C. robustum were characterized by phylogenetic analysis with corresponding genes from other berberine-producing species. The co-existence of genes from different plant families in the deepest branch subclade implies that the differentiation of berberine synthesis genes occurred early in the evolution of berberine-producing plants. Furthermore, the copy number increment of the berberine synthesis genes was detected at the species level.

Comparative Transcriptomics for Genes Related to Berberine and Berbamine Biosynthesis in Berberidaceae.

Berberine and berbamine are bioactive compounds of benzylisoquinoline alkaloids (BIAs) present in Berberis species. The contents of berbamine are 20 times higher than berberine in leaf tissues in three closely related species: Berberis koreana, B. thunbergii and B. amurensis. This is the first report on the quantification of berberine compared to the berbamine in the Berberis species. Comparative transcriptome analyses were carried out with mRNAs from the leaf tissues of the three-species. The comparison of the transcriptomes of B. thunbergii and B. amurensis to those of B. koreana, B. thunbergii showed a consistently higher number of differentially expressed genes than B. amurensis in KEGG and DEG analyses. All genes encoding enzymes involved in berberine synthesis were identified and their expressions were variable among the three species. There was a single copy of CYP80A/berbamunine synthase in B. koreana. Methyltransferases and cytochrome P450 mono-oxidases (CYPs) are key enzymes for BIA biosynthesis. The current report contains the copy numbers and other genomic characteristics of the methyltransferases and CYPs in Berberis species. Thus, the contents of the current research are valuable for molecular characterization for the medicinal utilization of the Berberis species.

Effect of disinfecting harvesting knives with sodium hypochlorite on soft rot infection of Kimchi cabbage.

This study evaluated the effect of sterilizing harvesting knives with sodium hypochlorite (NaOCl) on soft rot in Kimchi cabbage. Knives were infected with Pectobacterium carotovorum subsp. carotovorum (Pcc), sterilized with NaOCl (100, 200, and 300 mg/L), and used to cut Kimchi cabbage slices, which were incubated for 70 h in a 28 °C incubator. In Kimchi cabbage slices cut with a Pcc-inoculated knife without NaOCl sterilization, symptoms began to appear after 20 h, and approximately 60% of the cabbage slices were infected after 70 h of incubation. In contrast, in cabbage cut with a sterilized knife, soft rot symptoms were delayed, and they began to appear after 40 h of incubation in the 200 mg/L-treated. The expression levels of PG10, PG12-1, PG12-3, WRKY 33, MPK3, ACO1, and ACO2 were increased in infected plants, and NaOCl treatment decreased these expression levels. Transmission of soft rot can be minimized by disinfecting harvesting knives with 200 mg/L NaOCl.

Effect of germination environment on the biochemical compounds and anti-inflammatory properties of soybean cultivars.

In this study, we investigated changes in the isoflavone content, total phenolic content (TPC), total flavonoid content (TFC), antioxidant activities (DPPH, ABTS), and anti-inflammatory activities of small-seeded and large-seeded soybean cultivars during germination (light/dark conditions). Total isoflavone content was higher at the seed stage in large-seeded soybeans, while it increased after 7 days of germination in small-seeded soybeans, particularly in response to light conditions, under which they had high TPC, TFC, and antioxidant activities. In large-seeded soybeans, the germination environment did not significantly affect TFC or DPPH inhibition, whereas TPC and ABTS inhibition were high under dark germination conditions. Extracts of sprouts exhibited superior anti-inflammatory activities. Nitric oxide production was slightly lower in small-seeded and large-seeded soybeans germinated under light and dark conditions, respectively. Our findings indicate that germinated soybeans improved nutritionally, and that enhancement of bioactivity under different germination environments could contribute to the selection of appropriate soybean cultivars.

Nurturing the Student, Sustaining the Mission: 20 Years of the International/Inner-City/Rural Preceptorship Program.

BACKGROUND AND OBJECTIVES: Specialized medical school educational tracks aim to increase the primary care workforce. The International/Inner-City/Rural Preceptorship (I2CRP) Program is unique in addressing multiple communities, a large cohort and applying the Self Determination Theory framework. This study examined program impact by analyzing the numbers of graduates matched into primary care and practicing in medically underserved communities. METHODS: We compared the match list of I2CRP graduates between 2000 and 2017 (n=204) to non-I2CRP Virginia Commonwealth University School of Medicine (VCU SOM) graduates (n=3,037). We analyzed the matches into primary care, National Health Service Corps (NHSC) priority specialties, and NHSC priority plus general surgery. We searched a federal database to determine which graduates are practicing in workforce shortage areas. RESULTS: Many more I2CRP graduates matched to primary care (71.1%), compared to non-I2CRP graduates (38.2%; P<.001). Within primary care, I2CRP graduates matched to family medicine more frequently than non-I2CRP graduates (36.3% vs 8.4%). Eighteen percent of posttraining I2CRP graduates work in rural areas and 41% work in medically underserved areas. CONCLUSIONS: I2CRP graduates are more likely to match to family medicine and primary care. I2CRP curriculum nurtures new medical students' interest in primary care, and self-determination theory provides a framework to organize the program curriculum. The program's impact endures as evidenced by participants' continued work in underserved areas after residency. Increasing support for such programs may help address the primary care physician shortage in medically underserved areas.

Addressing Noncommunicable Disease in Dominican Republic: Barriers to Hypertension and Diabetes Care.

INTRODUCTION: Noncommunicable diseases (NCDs) significantly contribute to morbidity and mortality worldwide. During medical brigades in Santo Domingo, the Dominican Aid Society of Virginia (DASV) collects data to help understand the dynamics of NCD management. This study presents findings regarding resources and barriers to NCD treatment. Methods: A cross-sectional survey study was conducted in two communities (Los Mina and Paraiso) during the 2014 DASV summer brigade. Descriptive statistics, associations, correlations as well as qualitative analyses were conducted to better understand resources and barriers to care in relation to health care coverage. RESULTS: More than one third (n = 64) of 165 individuals had hypertension and/or diabetes. Thirty-seven percent (Paraiso) and 46% (Los Mina) of study participants did not have health insurance in the previous year. For those that did have insurance, 77% (P) and 89% (LM) visited a physician in the previous year. In this same group, 65% of individuals from Paraiso reported that their health insurance never covered the cost of medications while only a quarter of individuals from Los Mina indicated this. Health insurance and access to physicians and medication varied depending on the community of residence. Surveys indicated that access to affordable medications was an important issue for participants. Also, even though individuals in Los Mina were less likely to have health insurance than those in Paraiso, they were more likely to visit a physician. CONCLUSION: This study contributes to a greater understanding of health care coverage and access for low-resource communities in the Dominican Republic. Health care access, insurance, and cost sharing differed between these communities, but barriers to care were common. Future investigations could focus on qualitative differences in communities' health insurance coverages and development of interventions to address obstacles to care.

A Comparative Study of Phenolic Antioxidant Activity and Flavonoid Biosynthesis-Related Gene Expression Between Summer and Winter Strawberry Cultivars.

Strawberry (Fragaria ananassa Duch.) possesses good antioxidant properties. Phenolic compounds in strawberries, such as anthocyanins and ellagic acid, mainly act as antioxidants. This study aimed to compare the phenolic content and expression patterns of genes involved in flavonoid biosynthesis between summer and winter strawberry cultivars affected by seasonal variation, degree of ripeness, and genotype. Antioxidant activity and the total content of phenols and flavonoids decreased with fruit ripening. Most notably, summer strawberry cultivars showed higher antioxidant activity than winter cultivars. The expression patterns of flavonoid biosynthetic genes tested were cultivar-dependent and were also affected by ripening. These results help us understand the nutritional and physiological characteristics of selected cultivars and provide a range of information for strawberry consumption.

The SCAR/WAVE complex polarizes PAN receptors and promotes division asymmetry in maize.

Pre-mitotic establishment of polarity is a key event in the preparation of mother cells for asymmetric cell divisions that produce daughters of distinct fates, and ensures correct cellular patterning of tissues and eventual organ function. Previous work has shown that two receptor-like kinases, PANGLOSS2 (PAN2) and PAN1, and the small GTPase RHO GTPASE OF PLANTS (ROP) promote mother cell polarity and subsequent division asymmetry in developing maize stomata. PAN proteins become polarized prior to asymmetric cell division, however, the mechanism of this polarization is unknown. Here we show that the SCAR/WAVE regulatory complex, which activates the actin-nucleating ARP2/3 complex, is the first known marker of polarity in this asymmetric division model and is required for PAN polarization. These findings implicate actin, and specifically branched actin networks, in PAN polarization and asymmetric cell division.

Effects of ginsenoside Rb1 on the stress-induced changes of BDNF and HSP70 expression in rat hippocampus.

Ginsenoside Rb1 (GRb1) has been determined to exert diverse neuromodulatory effects including antistress effects in the brain. The hippocampus is a key brain structure for memory, learning, and cognition and is especially vulnerable to neurotoxic effects associated with stress. The aim of this study was to further explore neuroprotective potential of GRb1 on stress-mediated changes in hippocampal gene expression. Recent studies recognize agents that inducing brain-derived neurotrophic factor (BDNF) and heat shock protein (HSP) 70 as important neuroprotective approaches. Thus, we specifically determined the effects of GRb1 on mRNA expression of BDNF and HSP70, in a model of immobilization stress. In agreement with these reports, acute immobilization stress led to a decrease and an increase in the mRNA levels of the BDNF and HSP70, respectively, in the hippocampus. When pretreated orally, GRb1 significantly inhibited the stress-mediated decline of BDNF level whereas it further increased the stress-mediated elevation of HSP70 level. Our results strongly suggest GRb1 effective in controlling stress-related hippocampal dysfunction. Our finding also contributes further understanding of medicinal usefulness of GRb1 targeting hippocampal network alteration which is commonly observed in aging and neurodegenerative disorders.

Identification of PAN2 by quantitative proteomics as a leucine-rich repeat-receptor-like kinase acting upstream of PAN1 to polarize cell division in maize.

Mechanisms governing the polarization of plant cell division are poorly understood. Previously, we identified pangloss1 (PAN1) as a leucine-rich repeat-receptor-like kinase (LRR-RLK) that promotes the polarization of subsidiary mother cell (SMC) divisions toward the adjacent guard mother cell (GMC) during stomatal development in maize (Zea mays). Here, we identify pangloss2 (PAN2) as a second LRR-RLK promoting SMC polarization. Quantitative proteomic analysis identified a PAN2 candidate by its depletion from membranes of pan2 single and pan1;pan2 double mutants. Genetic mapping and sequencing of mutant alleles confirmed the identity of this protein as PAN2. Like PAN1, PAN2 has a catalytically inactive kinase domain and accumulates in SMCs at sites of GMC contact before nuclear polarization. The timing of polarized PAN1 and PAN2 localization is very similar, but PAN2 acts upstream because it is required for polarized accumulation of PAN1 but is independent of PAN1 for its own localization. We find no evidence that PAN2 recruits PAN1 to the GMC contact site via a direct or indirect physical interaction, but PAN2 interacts with itself. Together, these results place PAN2 at the top of a cascade of events promoting the polarization of SMC divisions, potentially functioning to perceive or amplify GMC-derived polarizing cues.