A rice seed-specific glycine-rich protein OsDOR1 interacts with GID1 to repress GA signaling and regulates seed dormancy.

Seed dormancy is an important agronomic trait under the control of complex genetic and environmental interactions, which have not been yet comprehensively understood. From the field screening of rice mutant library generated by a Ds transposable element, we identified a pre-harvest sprouting (PHS) mutant dor1. This mutant has a single insertion of Ds element at the second exon of OsDOR1 (LOC_Os03g20770), which encodes a novel seed-specific glycine-rich protein. This gene successfully complemented the PHS phenotype of dor1 mutant and its ectopic expression enhanced seed dormancy. Here, we demonstrated that OsDOR1 protein binds to the GA receptor protein, OsGID1 in rice protoplasts, and interrupts with the formation OsGID1-OsSLR1 complex in yeast cells. Co-expression of OsDOR1 with OsGID1 in rice protoplasts attenuated the GA-dependent degradation of OsSLR1, the key repressor of GA signaling. We showed the endogenous OsSLR1 protein level in the dor1 mutant seeds is significantly lower than that of wild type. The dor1 mutant featured a hypersensitive GA-response of α-amylase gene expression during seed germination. Based on these findings, we suggest that OsDOR1 is a novel negative player of GA signaling operated in the maintenance of seed dormancy. Our findings provide a novel source of PHS resistance.

Validation of a Modified Version of Interpersonal Reactivity Index for Medical Students Using Rasch Analysis.

Construct: Empathy has been accepted to interweave both cognitive aspects (the ability to put oneself in another person's place), and affective (or emotional) aspects, indicating an emotional reaction or response to another person's emotional state. Literature supports the positive influences of empathy on doctor-patient relationship, patient satisfaction, and positive clinical outcomes. Background: Many studies have dealt with the development of empathy measurement tools for physicians and medical students. A frequently used empathy measuring instrument for medical students is the "Interpersonal Reactivity Index" (IRI) which was designed to measure the multi-dimensional aspects of empathy in the general adult population. Most previous literature which validated IRI for medical students has used factor analysis, whilst studies applying Rasch models have been limited. Our study aimed to investigate the psychometric properties of a modified version of IRI for medical students using Rasch analysis. Approach: Medical students (1,293) from 15 medical schools in South Korea participated in an online questionnaire consisting of 28 items of the Korean translated version of IRI. We applied exploratory factor analysis (EFA) using polychoric correlation matrix to determine the optimal number of factors followed by Rasch analysis and McDonald's Omega calculation. Findings: The adapted IRI-MS (IRI for medical students) consisted of 17 items in four dimensions: empathic concern (5), fictitious situation (4), perspective taking (4), and personal distress (4). The overall fit of IRI-MS revealed an acceptable goodness-of-fit for all 17 items and a positive point measure correlation for all items. Reliability indices from the Rasch modeling and McDonald's Omega values of all four dimensions were satisfactory for research. We found the Wright-Andrich maps and category probability curves of the IRI's four dimensions to be less than optimal in measuring empathy levels with adequate precision. Conclusions: Rasch analysis of IRI-MS fell short from being able to prove satisfactory validity in measuring the multidimensional nature of empathy in medical students. However, our study applying Rasch analysis may serve as groundwork for future studies, to further develop from the shortcomings of our findings.Supplemental data for this article are available online at at www.tandfonline.com/htlm .

Unraveling the Mechanism and Practical Implications of the Sol-Gel Synthesis of Spinel LiMn2O4 as a Cathode Material for Li-Ion Batteries: Critical Effects of Cation Distribution at the Matrix Level.

Spinel LiMn2O4 (LMO) is a state-of-the-art cathode material for Li-ion batteries. However, the operating voltage and battery life of spinel LMO needs to be improved for application in various modern technologies. Modifying the composition of the spinel LMO material alters its electronic structure, thereby increasing its operating voltage. Additionally, modifying the microstructure of the spinel LMO by controlling the size and distribution of the particles can improve its electrochemical properties. In this study, we elucidate the sol-gel synthesis mechanisms of two common types of sol-gels (modified and unmodified metal complexes)-chelate gel and organic polymeric gel-and investigate their structural and morphological properties and electrochemical performances. This study highlights that uniform distribution of cations during sol-gel formation is important for the growth of LMO crystals. Furthermore, a homogeneous multicomponent sol-gel, necessary to ensure that no conflicting morphologies and structures would degrade the electrochemical performances, can be obtained when the sol-gel has a polymer-like structure and uniformly bound ions; this can be achieved by using additional multifunctional reagents, namely cross-linkers.

Burnout of Faculty Members of Medical Schools in Korea.

BACKGROUND: There is no national survey on medical school faculty members' burnout in Korea. This study aimed to investigate burnout levels and explore possible factors related to burnout among faculty members of Korean medical schools. METHODS: An anonymous online questionnaire was distributed to 40 Korean medical schools from October 2020 to December 2020. Burnout was measured by a modified and revalidated version of the Maslach Burnout Inventory-Human Service Survey. RESULTS: A total of 996 faculty members participated in the survey. Of them, 855 answered the burnout questions, and 829 completed all the questions in the questionnaire. A significant number of faculty members showed a high level of burnout in each sub-dimension: 34% in emotional exhaustion, 66.3% in depersonalization, and 92.4% in reduced personal accomplishment. A total of 31.5% of faculty members revealed a high level of burnout in two sub-dimensions, while 30.5% revealed a high level of burnout in all three sub-dimensions. Woman faculty members or those younger than 40 reported significantly higher emotional exhaustion and depersonalization. Long working hours (≥ 80 hours/week) showed the highest reduced personal accomplishment scores (F = 4.023, P = 0.018). The most significant stressor or burnout source was "excessive regulation by the government or university." The research was the most exasperating task, but the education was the least stressful. CONCLUSION: This first nationwide study alerts that a significant number of faculty members in Korean medical schools seem to suffer from a high level of burnout. Further studies are necessary for identifying the burnout rate, related factors, and strategies to overcome physician burnout.

Comparison of diagnostic performance of the ACR and Kwak TIRADS applying the ACR TIRADS' size thresholds for FNA.

OBJECTIVE: To investigate the diagnostic performances and unnecessary fine-needle aspiration (FNA) rates of two point-scale based TIRADS and compare them with a modified version using the ACR TIRADS' size thresholds. METHODS: Our Institutional Review Board approved this retrospective study and waived the requirement for informed consent. A total of 2106 thyroid nodules 10 mm or larger in size in 2084 patients with definitive cytopathologic findings were included. Ultrasonography categories were assigned according to each guideline. We applied the ACR TIRADS' size thresholds for FNA to the Kwak TIRADS and defined it as the modified Kwak TIRADS (mKwak TIRADS). Diagnostic performances and unnecessary FNA rates were evaluated for both the original and modified guidelines. RESULTS: Of the original guidelines, the ACR TIRADS had higher specificity, accuracy, and area under the receiver operating characteristic curve (AUC) (63.1%, 68.9%, and 0.748, respectively). When the size threshold of the ACR TIRADS was applied to the Kwak TIRADS, the resultant mKwak TIRADS had higher specificity, accuracy, and AUC (64.7%, 70.3%, and 0.765, respectively) than the ACR TIRADS. The mKwak TIRADS also had a lower unnecessary FNA rate than the ACR TIRADS (54.8% and 56.4%, respectively). The false-negative rate of the Kwak TIRADS was the lowest (1.9%) among all TIRADS. CONCLUSION: The modified Kwak TIRADS incorporating the size thresholds of the ACR TIRADS showed higher diagnostic performance and a lower unnecessary FNA rate than the original point-scale based TIRADS. KEY POINTS: • Of the original guidelines, the ACR TIRADS had the highest specificity, accuracy, and area under the receiver operating characteristic curve (AUC) (63.1%, 68.9%, and 0.748, respectively). • When the size threshold of the ACR TIRADS was applied to the Kwak TIRADS, the resultant modified version of Kwak TIRADS had higher specificity, accuracy, and AUC (64.7%, 70.3%, and 0.765, respectively) than the ACR TIRADS. • The false-negative rate of the Kwak TIRADS was the lowest (1.9%) among all TIRADS.

Molecular Identification of Anisakis Larvae Extracted by Gastrointestinal Endoscopy from Health Check-up Patients in Korea.

Anisakiasis is a zoonotic disease induced by anisakid nematodes, and endoscopic inspection is used for a diagnosis or remedy for it. Anisakis simplex, Anisakis physeteris, and Pseudoterranova decipiens had been reported to be the major species causing human infections, particularly, in Japan. However, in Korea, recent studies strongly suggested that Anisakis pegreffii is the major species of human infections. To support this suggestion, we collected anisakid larvae (n=20) from 20 human patients who were undergone gastrointestinal endoscopy at a health check-up center in Korea, and molecular identification was performed on the larvae using PCR-RFLP analysis and gene sequencing of rDNA ITS regions and mtDNA cox2. In addition, anisakid larvae (n=53) collected from the sea eel (Astroconger myriaster) were also examined for comparison with those extracted from humans. The results showed that all human samples (100%) were identified as A. pegreffii, whereas 90.7% of the samples from the sea eel were A. pegreffii with the remaining 9.3% being Hysterothylacium aduncum. Our study confirmed that A. pegreffii is the predominant species causing human anisakiasis in Korea, and this seems to be due to the predominance of this larval type in the fish (sea eels) popularly consumed by the Korean people. The possibility of human infection with H. aduncum in Korea is also suggested.

Molecular Genetic Findings of Spirometra decipiens and S. ranarum in Korea.

The taxonomy of Spirometra species has been controversial despite the medical and veterinary importance. Currently, only a few Spirometra species are considered valid species in the genus Spirometra. In the present study, the distribution of Spirometra species obtained from animals in Korea were identified by molecular analysis of the mitochondrial cytochrome c oxidase I (cox1) gene. A total of 28 Spirometra species specimens were analyzed. These were all collected between 1973 and 2008 in the Republic of Korea. Mitochondrial cox1 sequences were examined for a total of 28 specimens comprising 14 S. decipiens and 14 S. ranarum. The difference in partial cox1 sequences (316 bp) between S. erinaceieuropaei (KJ599680) and S. ranarum (this study) was 9.3%, while that between S. decipiens (KJ599679) and S. ranarum (this study) was 2.2%. Genetic analyses identified 2 Spirometra species in animals such as cat, leopard cat, dog, duck and snake in Korea as S. decipiens and S. ranarum. S. decipiens and S. ranarum were present in Gyeongnam Province (P), Jeonnam P, Gangwon P, Chungbuk P, and Seoul. S. decipiens was found in tadpoles, snakes, ducks, cats, leopard cats and dogs, while S. ranarum was found in cats and dogs. The ratio of S. decipiens:S. ranarum calculated from the molecular data was 14:14 (or 1:1). These results indicate that S. decipiens and S. ranarum are sympatrically distributed in Korea.

How do medical students differ in their interpersonal needs?

BACKGROUND: Knowing one's interpersonal relationship preferences can be tremendously helpful for medical students' lives. The purpose of this study was to examine the interpersonal needs in medical students. METHODS: Between 2010 and 2015, a total of 877 students from four Korean medical schools took the Korean version of the Fundamental Interpersonal Relations Orientation - Behaviour (FIRO-B) scale. The FIRO-B results were analyzed by descriptive statistics, frequency, independent t-test, and one-way ANOVA. RESULTS: The medical students' scores for interpersonal needs were moderate overall, with the highest scores for control (M = 8.63, SD = 3.08), followed by affection (M = 8.14, SD = 4.34), and inclusion (M = 7.81, SD = 4.30). Gender differences showed in three areas: expressed control (male > female, t = 4.137, p < 0.001), wanted affection (male < female, t = -3.148, p = 0.002), and control needs (male > female, t = 2.761, p = 0.006). By school type, differences were shown in expressed control (t = 3.581, p < 0.001), wanted inclusion (t = 2.625, p = 0.009), Inclusion (t = 1.966, p = 0.050), and expressed (t = 2.077, p = 0.038); undergraduate medical college (MC) students' needs were greater than the needs of graduate medical school (MS) students, but for wanted control, the MS students showed greater needs (t = -2.122, p = 0.034). CONCLUSIONS: There were differences in all categories except for expressed inclusion, wanted control, and control. The FIRO-B is a useful tool for giving insight into students regarding their interpersonal orientations, which will help them to adjust to medical school life. In addition, the FIRO-B can be useful when mentoring and coaching students.

Draft Genome of Toxocara canis, a Pathogen Responsible for Visceral Larva Migrans.

This study aimed at constructing a draft genome of the adult female worm Toxocara canis using next-generation sequencing (NGS) and de novo assembly, as well as to find new genes after annotation using functional genomics tools. Using an NGS machine, we produced DNA read data of T. canis. The de novo assembly of the read data was performed using SOAPdenovo. RNA read data were assembled using Trinity. Structural annotation, homology search, functional annotation, classification of protein domains, and KEGG pathway analysis were carried out. Besides them, recently developed tools such as MAKER, PASA, Evidence Modeler, and Blast2GO were used. The scaffold DNA was obtained, the N50 was 108,950 bp, and the overall length was 341,776,187 bp. The N50 of the transcriptome was 940 bp, and its length was 53,046,952 bp. The GC content of the entire genome was 39.3%. The total number of genes was 20,178, and the total number of protein sequences was 22,358. Of the 22,358 protein sequences, 4,992 were newly observed in T. canis. Following proteins previously unknown were found: E3 ubiquitin-protein ligase cbl-b and antigen T-cell receptor, zeta chain for T-cell and B-cell regulation; endoprotease bli-4 for cuticle metabolism; mucin 12Ea and polymorphic mucin variant C6/1/40r2.1 for mucin production; tropomodulin-family protein and ryanodine receptor calcium release channels for muscle movement. We were able to find new hypothetical polypeptides sequences unique to T. canis, and the findings of this study are capable of serving as a basis for extending our biological understanding of T. canis.

Cross-reactivity of Toxocariasis with Crude Antigen of Toxascaris leonina Larvae by ELISA.

Roundworms of Toxocara canis and Toxascaris leonina are common gastrointestinal helminths of canids over the world. Humans are infected with T. canis larvae through ingestion of infective eggs in contaminated environments or larvae by consumption of raw or uncooked meat or livers. Recently, patients of clinically diagnosed toxocariasis are increasing and require correct diagnosis in Korea. The present study investigated serological cross-reactivity between crude antigens of T. canis (TCLA) and T. leonina (TLLA) larvae. We collected serum specimens from 177 toxocariasis patients who were clinically suspected in the Seoul National University Hospital and 115 healthy controls. An ELISA method for toxocariasis was used to evaluate diagnostic efficacy of TLLA for serodiagnosis of human toxocariasis. The IgG ELISA using TLLA gave 14 (14.3%) positives of 98 TCLA positive specimens among 177 suspected toxocariasis patients. Most of them showed high absorbances with TCLA. In conclusion, there is a partial cross reaction between serum specimens of toxocariasis and TLLA.

Human Infections with Spirometra decipiens Plerocercoids Identified by Morphologic and Genetic Analyses in Korea.

Tapeworms of the genus Spirometra are pseudophyllidean cestodes endemic in Korea. At present, it is unclear which Spirometra species are responsible for causing human infections, and little information is available on the epidemiological profiles of Spirometra species infecting humans in Korea. Between 1979 and 2009, a total of 50 spargana from human patients and 2 adult specimens obtained from experimentally infected carnivorous animals were analyzed according to genetic and taxonomic criteria and classified as Spirometra erinaceieuropaei or Spirometra decipiens depending on the morphology. Morphologically, S. erinaceieuropaei and S. decipiens are different in that the spirally coiled uterus in S. erinaceieuropaei has 5-7 complete coils, while in S. decipiens it has only 4.5 coils. In addition, there is a 9.3% (146/1,566) sequence different between S. erinaceieuropaei and S. decipiens in the cox1 gene. Partial cox1 sequences (390 bp) from 35 Korean isolates showed 99.4% (388/390) similarity with the reference sequence of S. erinaceieuropaei from Korea (G1724; GenBank KJ599680) and an additional 15 Korean isolates revealed 99.2% (387/390) similarity with the reference sequences of S. decipiens from Korea (G1657; GenBank KJ599679). Based on morphologic and molecular databases, the estimated population ratio of S. erinaceieuropaei to S. decipiens was 35: 15. Our results indicate that both S. erinaceieuropaei and S. decipiens found in Korea infect humans, with S. erinaceieuropaei being 2 times more prevalent than S. decipiens. This study is the first to report human sparganosis caused by S. decipiens in humans in Korea.

Parasitic diseases as the cause of death of prisoners of war during the Korean War (1950-1953).

To determine the cause of death of prisoners of war during the Korean War (1950-1953), death certificates or medical records were analyzed. Out of 7,614 deaths, 5,013 (65.8%) were due to infectious diseases. Although dysentery and tuberculosis were the most common infectious diseases, parasitic diseases had caused 14 deaths: paragonimiasis in 5, malaria in 3, amoebiasis in 2, intestinal parasitosis in 2, ascariasis in 1, and schistosomiasis in 1. These results showed that paragonimiasis, malaria, and amoebiasis were the most fatal parasitic diseases during the early 1950s in the Korean Peninsula. Since schistosomiasis is not endemic to Korea, it is likely that the infected private soldier moved from China or Japan to Korea.

Serodiagnosis of toxocariasis by ELISA using crude antigen of Toxocara canis larvae.

Toxocariasis is a worldwide zoonosis caused by larvae of ascarid nematodes of dogs or cats, Toxocara canis or T. cati. Diagnosis of human toxocariasis currently relies on serology that uses T. canis excretory-secretory antigen to detect specific IgG antibodies by ELISA. We investigated the serodiagnostic efficacy of ELISA using crude antigen of T. canis larvae (TCLA). Serum specimens of 64 clinically confirmed toxocariasis, 115 healthy controls, and 119 other tissue-invading helminthiases were screened by ELISA using TCLA. The ELISA using TCLA showed 92.2% (59/64 patient samples) sensitivity and 86.6% (103/119) specificity. Its positive diagnostic predictivity was 78.7% and negative predictivity was 97.8%. No serum of healthy controls reacted but that of anisakiasis (45.5%), gnathostomiasis (19.2%), clonorchiasis (15.8%), sparganosis (11.1%), and cysticercosis (6.3%) cross-reacted. Immunoblot analysis on TCLA recognized antigenic proteins of 28- and 30-kDa bands in their dominant protein quantity and strong blotting reactivity. The present results indicate that the ELISA using our TCLA antigen is acceptable by the sensitivity and specificity for serodiagnosis of human toxocariasis. ELISA with TCLA is recommended to make differential diagnosis for patients with any sign of organ infiltration and eosinophilia.

Are ChatGPT's knowledge and interpretation ability comparable to those of medical students in Korea for taking a parasitology examination?: a descriptive study

This study aimed to compare the knowledge and interpretation ability of ChatGPT, a language model of artificial general intelligence, with those of medical students in Korea by administering a parasitology examination to both ChatGPT and medical students. The examination consisted of 79 items and was administered to ChatGPT on January 1, 2023. The examination results were analyzed in terms of ChatGPT's overall performance score, its correct answer rate by the items' knowledge level, and the acceptability of its explanations of the items. ChatGPT's performance was lower than that of the medical students, and ChatGPT's correct answer rate was not related to the items' knowledge level. However, there was a relationship between acceptable explanations and correct answers. In conclusion, ChatGPT's knowledge and interpretation ability for this parasitology examination were not yet comparable to those of medical students in Korea.

Journal metrics, document network, and conceptual and social structures of the Korean Journal of Anesthesiology from 2017 to July 2022: a bibliometric study.

BACKGROUND: This study aimed to identify the directions of research published in the Korean Journal of Anesthesiology (KJA) and identify the main topics and journal network through a bibliometric analysis. The results can be reflected in strategies for the journal's promotion to a top-ranking journal in the anesthesiology category. METHODS: KJA articles from January 1, 2017 to September 11, 2022 were retrieved from the Web of Science Core Collection on September 11, 2022, and analyzed using Biblioshiny. Journal metrics, the document network, the conceptual structure, and social structures were elucidated. RESULTS: Out of 525 articles, fewer than half (48.6%) were from Korean corresponding authors. The impact factor steeply increased from 2.316 in 2019 to 5.167 in 2021. The Hirsch index of KJA was 24. A co-occurrence network of Keywords Plus showed four clusters of central keywords: surgery, management, anesthesia, and mortality. The conceptual structure map of Keywords Plus showed a main cluster of anesthesia and analgesia, while another minor cluster included intubation and induction. The co-citation network demonstrated that KJA was in the same cluster of anesthesiology journals. The collaboration network of the authors' countries showed that Korean authors collaborated mainly with researchers in the United States and Canada. CONCLUSIONS: These results show KJA's developmental process of promotion to a top-tier journal in the anesthesiology category. Furthermore, the following strategies are suggested for journal promotion: recruitment of articles on emerging and highly citable topics; and more active collaboration of society members with researchers worldwide.

Recent Issues in Medical Journal Publishing and Editing Policies: Adoption of Artificial Intelligence, Preprints, Open Peer Review, Model Text Recycling Policies, Best Practice in Scholarly Publishing 4th Version, and Country Names in Titles.

In Korea, many editors of medical journal are also publishers; therefore, they need to not only manage peer review, but also understand current trends and policies in journal publishing and editing. This article aims to highlight some of these policies with examples. First, the use of artificial intelligence tools in journal publishing has increased, including for manuscript editing and plagiarism detection. Second, preprint publications, which have not been peer-reviewed, are becoming more common. During the COVID-19 pandemic, medical journals have been more willing to accept preprints to adjust rapidly changing pandemic health issues, leading to a significant increase in their use. Third, open peer review with reviewer comments is becoming more widespread, including the mandatory publication of peer-reviewed manuscripts with comments. Fourth, model text recycling policies provide guidelines for researchers and editors on how to appropriately recycle text, for example, in the background section of the Introduction or the Methods section. Fifth, journals should take into account the recently updated 4th version of the Principles of Transparency and Best Practice in Scholarly Publishing, released in 2022. This version includes more detailed guidelines on journal websites, peer review processes, advisory boards, and author fees. Finally, it recommends that titles of human studies include country names to clarify the cultural context of the research. Each editor must decide whether to adopt these six policies for their journals. Editor-publishers of society journals are encouraged to familiarize themselves with these policies so that they can implement them in their journals as appropriate.

A case of vocal cord gnathostomiasis diagnosed with sectional morphologies in a histopathological specimen from a Chinese woman living in Korea.

This study aimed to describe a rare case of gnathostomiasis in the vocal cord. A 54-year-old Chinese woman living in Korea visited with a chief complaint of voice change at the outpatient department of otorhinolaryngology in Hallym Sacred Heart Hospital, Hallym University on August 2, 2021. She had eaten raw conger a few weeks before the voice change developed, but her medical history and physical examinations demonstrated neither gastrointestinal symptoms nor other health problems. A round and red cystic lesion, recognized in the anterior part of the right vocal cord, was removed using forceps and scissors through laryngeal microsurgery. The histopathological specimen of the cyst revealed 3 cross-sections of a nematode larva in the lumen of the cyst wall composed of inflammatory cells and fibrotic tissues. They differ in diameter, from 190 µm to 235 µm. They showed characteristic cuticular layers with tegumental spines, somatic muscle layers, and gastrointestinal tracts such as the esophagus and intestine. Notably, intestinal sections consisted of 27-28 lining cells containing 0-4 nuclei per cell. We tentatively identified the nematode larva recovered from the vocal cord cystic lesion as the third-stage larva of Gnathostoma, probably G. nipponicum or G. hispidum, based on the sectional morphologies.

Effect of structural variation in the promoter region of RsMYB1.1 on the skin color of radish taproot.

Accumulation of anthocyanins in the taproot of radish is an agronomic trait beneficial for human health. Several genetic loci are related to a red skin or flesh color of radish, however, the functional divergence of candidate genes between non-red and red radishes has not been investigated. Here, we report that a novel genetic locus on the R2 chromosome, where RsMYB1.1 is located, is associated with the red color of the skin of radish taproot. A genome-wide association study (GWAS) of 66 non-red-skinned (nR) and 34 red-skinned (R) radish accessions identified three nonsynonymous single nucleotide polymorphisms (SNPs) in the third exon of RsMYB1.1. Although the genotypes of SNP loci differed between the nR and R radishes, no functional difference in the RsMYB1.1 proteins of nR and R radishes in their physical interaction with RsTT8 was detected by yeast-two hybrid assay or in anthocyanin accumulation in tobacco and radish leaves coexpressing RsMYB1.1 and RsTT8. By contrast, insertion- or deletion-based GWAS revealed that one large AT-rich low-complexity sequence of 1.3-2 kb was inserted in the promoter region of RsMYB1.1 in the nR radishes (RsMYB1.1nR), whereas the R radishes had no such insertion; this represents a presence/absence variation (PAV). This insertion sequence (RsIS) was radish specific and distributed among the nine chromosomes of Raphanus genomes. Despite the extremely low transcription level of RsMYB1.1nR in the nR radishes, the inactive RsMYB1.1nR promoter could be functionally restored by deletion of the RsIS. The results of a transient expression assay using radish root sections suggested that the RsIS negatively regulates the expression of RsMYB1.1nR, resulting in the downregulation of anthocyanin biosynthesis genes, including RsCHS, RsDFR, and RsANS, in the nR radishes. This work provides the first evidence of the involvement of PAV in an agronomic trait of radish.

Identification of a putative nuclear export signal motif in human NANOG homeobox domain.

NANOG is a homeobox-containing transcription factor that plays an important role in pluripotent stem cells and tumorigenic cells. To understand how nuclear localization of human NANOG is regulated, the NANOG sequence was examined and a leucine-rich nuclear export signal (NES) motif ((125)MQELSNILNL(134)) was found in the homeodomain (HD). To functionally validate the putative NES motif, deletion and site-directed mutants were fused to an EGFP expression vector and transfected into COS-7 cells, and the localization of the proteins was examined. While hNANOG HD exclusively localized to the nucleus, a mutant with both NLSs deleted and only the putative NES motif contained (hNANOG HD-ΔNLSs) was predominantly cytoplasmic, as observed by nucleo/cytoplasmic fractionation and Western blot analysis as well as confocal microscopy. Furthermore, site-directed mutagenesis of the putative NES motif in a partial hNANOG HD only containing either one of the two NLS motifs led to localization in the nucleus, suggesting that the NES motif may play a functional role in nuclear export. Furthermore, CRM1-specific nuclear export inhibitor LMB blocked the hNANOG potent NES-mediated export, suggesting that the leucine-rich motif may function in CRM1-mediated nuclear export of hNANOG. Collectively, a NES motif is present in the hNANOG HD and may be functionally involved in CRM1-mediated nuclear export pathway.

Regulation of OCT4 gene expression by liver receptor homolog-1 in human embryonic carcinoma cells.

We demonstrate the regulation of OCT4 gene expression mediated by liver receptor homolog-1 (LRH-1) in human embryonic carcinoma cells. LRH-1 and OCT4 are co-expressed in undifferentiated NCCIT cells and decreased during retinoic acid-induced differentiation. Dose-dependent overexpression of LRH-1 transactivated the OCT4 promoter activity and its dominant negative form with a deletion of activation function-2 motif reduced the activity even in the presence of LRH-1. The OCT4 promoter contains potent three LRH-1 binding sites; one within conserved region (CR) 1 and two within CR2. Mutagenesis of each binding site affected the decrease in OCT4 promoter activity and the 2nd binding site mutant most significantly reduced the transcriptional activity, compared to that of 1st and 3rd binding site mutants, respectively. Simultaneous disruption of 2nd and 3rd binding sites led to significant down-regulation of the activity even in the presence of 1st binding site-containing CR1. Moreover, mutation of each binding element within native or exogenous minimal promoter-driven CR1 or CR2 also decreased the promoter activity to some different extent, suggesting that three binding elements may be implicated in the induction of the full-activity of OCT4 promoter. In vivo binding assay revealed the 2nd and 3rd binding motifs within CR2 were more enriched than the 1st one within CR1. Taken together, our study indicates that LRH-1 acts as a transcriptional activator in the regulation of OCT4 gene expression through the cooperative interaction with three binding sites directly or/and indirectly.