High-Throughput Phenotyping Approach for the Evaluation of Heat Stress in Korean Ginseng (Panax ginseng Meyer) Using a Hyperspectral Reflectance Image.

Panax ginseng has been used as a traditional medicine to strengthen human health for centuries. Over the last decade, significant agronomical progress has been made in the development of elite ginseng cultivars, increasing their production and quality. However, as one of the significant environmental factors, heat stress remains a challenge and poses a significant threat to ginseng plants' growth and sustainable production. This study was conducted to investigate the phenotype of ginseng leaves under heat stress using hyperspectral imaging (HSI). A visible/near-infrared (Vis/NIR) and short-wave infrared (SWIR) HSI system were used to acquire hyperspectral images for normal and heat stress-exposed plants, showing their susceptibility (Chunpoong) and resistibility (Sunmyoung and Sunil). The acquired hyperspectral images were analyzed using the partial least squares-discriminant analysis (PLS-DA) technique, combining the variable importance in projection and successive projection algorithm methods. The correlation of each group was verified using linear discriminant analysis. The developed models showed 12 bands over 79.2% accuracy in Vis/NIR and 18 bands with over 98.9% accuracy at SWIR in validation data. The constructed beta-coefficient allowed the observation of the key wavebands and peaks linked to the chlorophyll, nitrogen, fatty acid, sugar and protein content regions, which differentiated normal and stressed plants. This result shows that the HSI with the PLS-DA technique significantly differentiated between the heat-stressed susceptibility and resistibility of ginseng plants with high accuracy.

Homology threading to generate RNA polymerase structures.

Homology threading is a powerful technology for generating structural models based on homologous structures. Here we use threading to generate four complex RNA polymerase models. The models appear to be as useful as x-ray crystal structures or cryo-electron microscopy structures to support research projects.

Type-II tRNAs and Evolution of Translation Systems and the Genetic Code.

Because tRNA is the core biological intellectual property that was necessary to evolve translation systems, tRNAomes, ribosomes, aminoacyl-tRNA synthetases, and the genetic code, the evolution of tRNA is the core story in evolution of life on earth. We have previously described the evolution of type-I tRNAs. Here, we use the same model to describe the evolution of type-II tRNAs, with expanded V loops. The models are strongly supported by inspection of typical tRNA diagrams, measuring lengths of V loop expansions, and analyzing the homology of V loop sequences to tRNA acceptor stems. Models for tRNA evolution provide a pathway for the inanimate-to-animate transition and for the evolution of translation systems, the genetic code, and cellular life.

A novel inversion in the chloroplast genome of marama (Tylosema esculentum).

Tylosema esculentum (marama bean) is being developed as a possible crop for resource-poor farmers in arid regions of Southern Africa. As part of the molecular characterization of this species, the chloroplast genome has been assembled from next-generation sequencing using both Illumina and Pac-Bio data. The genome is of typical organization with a large single-copy region and a small single-copy region separated by a pair of inverted repeats and covers 161537 bp. It contains a unique inversion not present in any other legumes, even in the closest relatives for which the complete chloroplast genome is available, and two complete copies of the ycf1 gene. These data extend the range of variability of legume chloroplast genomes. The sequencing of multiple individuals has identified two different chloroplast genomes which were geographically separated. The current sampling is limited so that the extent of the intraspecific variation is still to be determined, leaving open the question of legume chloroplast genomes adapted to particular arid environments.

Opting out: confidentiality and availability of an 'alibi' for potential living kidney donors in the USA.

Ethicists and guidelines have suggested that potential living kidney donors who withdraw from evaluation be offered an 'alibi.' We sought to determine what potential living kidney donors are told about their ability to opt out, alibi availability and postwithdrawal confidentiality. We reviewed 148 consent forms for living kidney donor evaluation from US transplant centres that performed >5 living kidney transplants in 2010-2011 (response rate 87%). We found that while 98% of centres used evaluation consent forms that indicated that the donor could withdraw, only 21% of these documents offered an alibi. Another 23% of centres' consent forms indicated that the transplant team would be willing to inform the intended recipient that an individual was not a potential donor. Relatively few consent documents explicitly addressed the confidentiality of the donor's health information (31%), candidacy status (18%), decision (24%) or reasons (23%) following withdrawal. To preserve potential donors' autonomy and relationships, we advocate that all transplant centres offer general alibis in their evaluation consent forms. We conclude by offering recommendations for evaluation consent discussions of opting out, alibis and postwithdrawal confidentiality.

Conserved tyrosine kinase promotes the import of silencing RNA into Caenorhabditis elegans cells.

RNA silencing in Caenorhabditis elegans is transmitted between cells by the transport of double-stranded RNA (dsRNA). The efficiency of such transmission, however, depends on both the cell type and the environment. Here, we identify systemic RNAi defective-3 (SID-3) as a conserved tyrosine kinase required for the efficient import of dsRNA. Without SID-3, cells perform RNA silencing well but import dsRNA poorly. Upon overexpression of SID-3, cells import dsRNA more efficiently than do wild-type cells and such efficient import of dsRNA requires an intact SID-3 kinase domain. The mammalian homolog of SID-3, activated cdc-42-associated kinase (ACK), acts in many signaling pathways that respond to environmental changes and is known to directly associate with endocytic vesicles, which have been implicated in dsRNA transport. Therefore, our results suggest that the SID-3/ACK tyrosine kinase acts as a regulator of RNA import into animal cells.

Written informed consent for living liver donor evaluation: compliance with Centers for Medicare and Medicaid Services and Organ Procurement and Transplantation Network Guidelines and alibi offers.

We examined written informed consent forms for living liver donor evaluations to determine whether they incorporated elements required by the Centers for Medicare and Medicaid Services (CMS) and suggested by the Organ Procurement and Transplantation Network (OPTN). We contacted each of the 41 US centers that performed at least 1 living donor liver transplant in 2011; 37 centers reported active living donor evaluation programs. Twenty-six centers shared their consent form for living donor evaluation (response rate = 70%). Each document was double-coded for consent element content. We found that 57% of the centers included the 9 mandated CMS elements. Although the OPTN guidelines are non-binding, 78% of the centers used consent forms that addressed at least two-thirds of the elements recommended by OPTN. Only 17% of the centers provided written offers of an alibi to donors who withdrew from the evaluation. On the basis of our findings, we offer suggestions that may be relevant to ongoing revisions to the OPTN living liver donor consent policy and may help centers to improve the clarity of their written consent forms.

Hypericins: biotechnological production from cell and organ cultures.

Hypericum perforatum L. (St. John's wort), a perennial flowering plant native to Europe, is widely used as a medicinal plant and has a long history of its use in the treatment of various ailments. Currently, H. perforatum is widely used as an herbal remedy for the treatment of mild to moderate depression. Hypericins are natural napthodianthrone compounds produced from H. perforatum (St. John's wort) which are having antitumor, antiviral (i.e., against human immunodeficiency and hepatitis C virus), antineoplastic, and antidepressant properties. Currently, field-grown plant materials are generally used for the commercial production of hypericins. It has been reported that hypericin accumulation in natural plants is influenced by different ecological and environmental conditions including light intensity, nitrogen availability, temperature, seasons, and growing regions. Therefore, up to 17-fold and 13-fold differences in hypericin and pseudohypericin amounts, respectively, are reported in different phytopharmaceutical preparations. Plant cell and organ cultures are effective systems for producing natural products, and attempts were made for the production of biomass and stable concentrations of hypericins through in vitro cultures of H. perforatum. Cell, callus, shoot, plantlet, and adventitious root cultures have been established and various chemical and physical factors which influence the biomass and secondary metabolite accumulation have been investigated. Large-scale plantlet and adventitious root cultures have also been attempted in H. perforatum in bioreactors, and various strategies have been applied for the production of higher biomass and secondary products. This review describes the biotechnological approaches employed for the production of hypericins and focuses upon the challenges and future prospects.

Biotechnological production of caffeic acid derivatives from cell and organ cultures of Echinacea species.

Caffeic acid derivatives (CADs) are a group of bioactive compounds which are produced in Echinacea species especially Echinacea purpurea, Echinacea angustifolia, and Echinacea pallida. Echinacea is a popular herbal medicine used in the treatment of common cold and it is also a prominent dietary supplement used throughout the world. Caffeic acid, chlorogenic acid (5-O-caffeoylquinic acid), caftaric acid (2-O-caffeoyltartaric acid), cichoric acid (2, 3-O-dicaffeoyltartaric acid), cynarin, and echinacoside are some of the important CADs which have varied pharmacological activities. The concentrations of these bioactive compounds are species specific and also they vary considerably with the cultivated Echinacea species due to geographical location, stage of development, time of harvest, and growth conditions. Due to these reasons, plant cell and organ cultures have become attractive alternative for the production of biomass and caffeic acid derivatives. Adventitious and hairy roots have been induced in E. pupurea and E. angustifolia, and suspension cultures have been established from flask to bioreactor scale for the production of biomass and CADs. Tremendous progress has been made in this area; various bioprocess methods and strategies have been developed for constant high-quality productivity of biomass and secondary products. This review is aimed to discuss biotechnological methods and approaches employed for the sustainable production of CADs.

Biotechnological production of eleutherosides: current state and perspectives.

Eleutherosides, the phenylpropanoid and lignan glycosides, are the active ingredients accumulated in the roots and stems of Eleutherococcus species and in Eleutherococcus senticosus in particular. Syringin (=eleutheroside B) and (-) syringaresinol-di-O-ß-D-glucoside (=eleutheroside E) appear as the most important bioactive compounds which are used as adaptogens, besides their abundant antidiabetic and anticancer properties. As the availability of "Eleuthero" is becoming increasingly limited because of its scanty natural distribution, the production of these compounds by biotechnological means has become an attractive alternative. In E. senticosus and other closely related species, Eleutherococcus sessiliflorus, Eleutherococcus chiisanensis, and Eleutherococcus koreanum, organogenic cultures have been induced for the production of eleutherosides. Bioreactor cultures have been established and various parameters, which influence on the accumulation of biomass and secondary metabolites, have been thoroughly investigated. Pilot-scale cultures have also been accomplished for the large-scale production of somatic embryos containing abundant amounts of eleutherosides. This review describes the biotechnological approaches and challenges for the production of eleutherosides.

Ginsenosides: prospective for sustainable biotechnological production.

Panax ginseng C.A. Meyer (ginseng) is a well-known medicinal plant that has been traditionally used in the oriental countries for centuries. Wild ginseng is a scarce and rare commodity. Field cultivation of the ginseng plant is a time-consuming and labor-intensive process. Ginsenosides, a group of glycosylated triterpenes, also known as saponins, are the principal bioactive constituents of ginseng. The use of cell and organ culture processes has been sought as a potential alternative for the efficient mass production of ginseng raw material. Various bioprocessing strategies have been developed to date. Cells and adventitious roots have been cultured in large-scale bioreactors and various strategies have been developed accordingly for the enhancement of biomass and ginsenoside accumulation. This review highlights the recent progress in the cultivation of ginseng cell and organ cultures for the production of ginsenosides from bioreactor cultures. In addition, the metabolism and biochemistry of ginsenoside biosynthesis, genomic and proteomic studies in ginseng, metabolic engineering, biosafety, toxicological evaluation, and efficacy assessment of ginseng raw material are also summarized and thoroughly discussed.

Demineralized bone-fibrin sandwich for genioplasty.

UNLABELLED: Osseous genioplasty is a versatile technique to improve chin and facial aesthetics. It carries several benefits compared to an alloplastic implant, but the downsides of an intersegmental gap and lateral stepoffs must be weighed. These areas are typically left open, despite being subject to unpredictable resorption or incomplete osseous healing. We describe the strategy of using a combination of demineralized bone matrix (DBM) and fibrin glue to promote bone fill while smoothing contours and transitions along the genioplasty. Our retrospective study included 32 patients who underwent genioplasty with the DBM-fibrin sandwich. There were no genioplasty-related complications at the 6 months follow-up. Our results indicate that this technique is safe and is a valuable addition to possible graft material for genioplasty. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors at www.springer.com/00266 .

Tobacco NtLTP1, a glandular-specific lipid transfer protein, is required for lipid secretion from glandular trichomes.

Glandular trichomes are the phytochemical factories of plants, and they secrete a wide range of commercially important natural products such as lipids, terpenes and flavonoids. Herein, we report that the Nicotiana tabacum LTP1 (NtLTP1) gene, which is specifically expressed in long glandular trichomes, plays a role in lipid secretion from trichome heads. NtLTP1 mRNA is abundantly transcribed in trichomes, but NtLTP3, NtLTP4 and NtLTP5 are not. In situ hybridization revealed that NtLTP1 mRNAs accumulate specifically in long trichomes and not in short trichomes or epidermal cells. X-gluc staining of leaves from a transgenic plant expressing the NtLTP1 promoter fused to a GUS gene revealed that NtLTP1 protein accumulated preferentially on the tops of long glandular trichomes. GFP fluorescence from transgenic tobacco plants expressing an NtLTP1-GFP fusion protein was localized at the periphery of cells and in the excreted liquid droplets from the glandular trichome heads. In vitro assays using a fluorescent 2-p-toluidinonaphthalene-6-sulfonate probe indicated that recombinant NtLTP1 had lipid-binding activity. The overexpression of NtLTP1 in transgenic tobacco plants resulted in the increased secretion of trichome exudates, including epicuticular wax. In transgenic NtLTP1-RNAi lines, liquid secretion from trichomes was strongly reduced, but epicuticular wax secretion was not altered. Moreover, transgenic tobacco plants overexpressing NtLTP1 showed increased protection against aphids. Taken together, these data suggest that NtLTP1 is abundantly expressed in long glandular trichomes, and may play a role in lipid secretion from long glandular trichomes.

Helping science and drug development to succeed through pharma-academia partnerships: Yale Healthcare Conference 2013.

The theme of the 2013 Yale Healthcare Conference was "Partnerships in Healthcare: Cultivating Collaborative Solutions." The April conference brought together leaders across several sectors of health care, including academic research, pharmaceuticals, information technology, policy, and life sciences investing. In particular, the breakout session titled "Taking R&D Back to School: The Rise of Pharma-Academia Alliances" centered on the partnerships between academic institutions and pharmaceutical companies. Attendees of the session included members of the pharmaceutical industry, academic researchers, and physicians, as well as graduate and professional students. The discussion was led by Dr. Thomas Lynch of Yale University. Several topics emerged from the discussion, including resources for scientific discovery and the management of competing interests in collaborations between academia and the pharmaceutical industry.

Factors influencing COVID-19 vaccination intention among parents of children aged 5-11 years in South Korea: a cross-sectional study.

PURPOSE: The purpose of this study was to identify factors affecting parents' intentions to have their children aged 5-11 years vaccinated against coronavirus disease 2019 (COVID-19). METHODS: The participants of the study were 298 parents with children aged 5-11 years in South Korea. Data collection took place from October 20 to October 26, 2022 and used an online survey (Google Forms). Data were analyzed using descriptive statistics, the t test, analysis of variance, the Scheffé test, Pearson correlation coefficients, and hierarchical regression in IBM SPSS version 26.0. RESULTS: The factors influencing participants' vaccination intentions for their children aged 5-11 years were cognitive behavioral control (ß=.40, p<.001), attitudes (ß=.37, p<.001), subjective norms (ß=.20, p<.001), and awareness of whether their child could receive the COVID-19 vaccine (ß=.07, p=.016). The explanatory power of the regression equation was 89%. CONCLUSION: Parents' intentions to vaccinate their children against COVID-19 are influenced by their attitudes, subjective norms, and perceived behavioral control toward vaccines. Since parents are concerned about vaccine side effects, it is important to establish a trusted line of communication to keep them informed about vaccinations.

Factors influencing the learning transfer of nursing students in a non-face-to-face educational environment during the COVID-19 pandemic in Korea: a cross-sectional study using structural equation modeling.

PURPOSE: The aim of this study was to identify factors influencing the learning transfer of nursing students in a non-face-to-face educational environment through structural equation modeling and suggest ways to improve the transfer of learning. METHODS: In this cross-sectional study, data were collected via online surveys from February 9 to March 1, 2022, from 218 nursing students in Korea. Learning transfer, learning immersion, learning satisfaction, learning efficacy, self-directed learning ability and information technology utilization ability were analyzed using IBM SPSS for Windows ver. 22.0 and AMOS ver. 22.0. RESULTS: The assessment of structural equation modeling showed adequate model fit, with normed χ2=1.74 (P<0.024), goodness-of-fit index=0.97, adjusted goodness-of-fit index=0.93, comparative fit index=0.98, root mean square residual=0.02, Tucker-Lewis index=0.97, normed fit index=0.96, and root mean square error of approximation=0.06. In a hypothetical model analysis, 9 out of 11 pathways of the hypothetical structural model for learning transfer in nursing students were statistically significant. Learning self-efficacy and learning immersion of nursing students directly affected learning transfer, and subjective information technology utilization ability, self-directed learning ability, and learning satisfaction were variables with indirect effects. The explanatory power of immersion, satisfaction, and self-efficacy for learning transfer was 44.4%. CONCLUSION: The assessment of structural equation modeling indicated an acceptable fit. It is necessary to improve the transfer of learning through the development of a self-directed program for learning ability improvement, including the use of information technology in nursing students' learning environment in non-face-to-face conditions.

Intra-person multi-task learning method for chronic-disease prediction.

In the medical field, various clinical information has been accumulated to help clinicians provide personalized medicine and make better diagnoses. As chronic diseases share similar characteristics, it is possible to predict multiple chronic diseases using the accumulated data of each patient. Thus, we propose an intra-person multi-task learning framework that jointly predicts the status of correlated chronic diseases and improves the model performance. Because chronic diseases occur over a long period and are affected by various factors, we considered features related to each chronic disease and the temporal relationship of the time-series data for accurate prediction. The study was carried out in three stages: (1) data preprocessing and feature selection using bidirectional recurrent imputation for time series (BRITS) and the least absolute shrinkage and selection operator (LASSO); (2) a convolutional neural network and long short-term memory (CNN-LSTM) for single-task models; and (3) a novel intra-person multi-task learning CNN-LSTM framework developed to predict multiple chronic diseases simultaneously. Our multi-task learning method between correlated chronic diseases produced a more stable and accurate system than single-task models and other baseline recurrent networks. Furthermore, the proposed model was tested using different time steps to illustrate its flexibility and generalization across multiple time steps.

Hyperspectral reflectance imaging for nondestructive evaluation of root rot in Korean ginseng (Panax ginseng Meyer).

Root rot of Panax ginseng caused by Cylindrocarpon destructans, a soil-borne fungus is typically diagnosed by frequently checking the ginseng plants or by evaluating soil pathogens in a farm, which is a time- and cost-intensive process. Because this disease causes huge economic losses to ginseng farmers, it is important to develop reliable and non-destructive techniques for early disease detection. In this study, we developed a non-destructive method for the early detection of root rot. For this, we used crop phenotyping and analyzed biochemical information collected using the HSI technique. Soil infected with root rot was divided into sterilized and infected groups and seeded with 1-year-old ginseng plants. HSI data were collected four times during weeks 7-10 after sowing. The spectral data were analyzed and the main wavelengths were extracted using partial least squares discriminant analysis. The average model accuracy was 84% in the visible/near-infrared region (29 main wavelengths) and 95% in the short-wave infrared (19 main wavelengths). These results indicated that root rot caused a decrease in nutrient absorption, leading to a decline in photosynthetic activity and the levels of carotenoids, starch, and sucrose. Wavelengths related to phenolic compounds can also be utilized for the early prediction of root rot. The technique presented in this study can be used for the early and timely detection of root rot in ginseng in a non-destructive manner.

Dammarenediol-II production confers TMV tolerance in transgenic tobacco expressing Panax ginseng dammarenediol-II synthase.

Panax ginseng is one of the famous medicinal plants. Ginsenosides, a class of tetracyclic triterpene saponins, are mainly responsible for its pharmacological activity. Most ginsenosides are composed of dammarenediol-II aglycone with various sugar moieties. Dammarenediol-II synthase is the first enzyme in the biosynthesis of ginsenosides. Here, we report that transgenic tobacco expressing the P. ginseng dammarenediol-II synthase gene (PgDDS) produced dammarenediol-II, and conferred resistance to Tobacco mosaic virus (TMV). Upon infection with TMV, lesions developed more rapidly in transgenic tobacco plants, and their size was smaller than those of wild-type plants. Transgenic tobacco plants showed a low level of both the viral titer and mRNA accumulation of TMV coat protein (CP) compared with the wild type. The production of dammarenediol-II in transgenic tobacco stimulated the expression of tobacco pathogenesis-related genes (PR1 and PR2) under both virus-untreated and -treated conditions. When the leaves of wild-type plants were inoculated with a mixture of TMV and dammarenediol-II, the leaves exhibited a reduced viral concentration and TMV-CP expression than those receiving TMV treatment alone. When the leaves of P. ginseng were infected with TMV, transcription of PgDDS was significantly increased. Transgenic P. ginseng plants harboring a ß-glucuronidase (GUS) gene driven by the PgDDS promoter were constructed. The GUS expression was activated when the transgenic ginseng plants were treated with TMV. These results indicate that the medicinally important dammarenediol-II can be ectopically produced in tobacco, and the production of dammarenediol-II in tobacco plants allows them to adopt a viral defense system.

Positive Effects on Emotional Stress and Sleep Quality of Forest Healing Program for Exhausted Medical Workers during the COVID-19 Outbreak.

This study targeted medical workers, who are currently being subjected to an excessive workload and emotional stress during the COVID-19 outbreak. Various treatment programs, such as a relaxation program to relieve stress, a walk in the forest, and woodworking were provided to the participants as forest healing therapies. We enrolled 13 medical workers (11 females, 2 males). Before and after forest healing therapy, stress and sleep-related questionnaires and levels of salivary cortisol, dehydroepiandrosterone sulfate (DHEA-S), and melatonin were measured and compared. The improvement of the perceived stress scale and the decrease of DHEA-S, a stress index, showed statistically significant results. However, although this study was conducted with a small number of participants and has a limitation in that the therapy occurred over a short period of only 1 night and 2 days, the trend of supporting results remains positive. As such, the authors propose forest healing therapy as one intervention to relieve the job stress for this group of workers.