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The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), remained a major global health concern in 2021. To suppress the spread of infection, mass vaccinations have been performed across countries worldwide. In Japan, vaccinations of the first and second doses for most of the nation were performed during the nationwide outbreak of the B.1.617.2 (Delta) variant with the L452R spike protein mutation, and the effectiveness of the vaccinations to suppress the spread of COVID-19 among the people in Japan remains uncertain. In this study, adults aged ≥18 years, who were in contact with patients with COVID-19 and underwent nasopharyngeal swab reverse transcription-polymerase chain reaction (RT-PCR) tests during August and September 2021 at a mass screening test center in Japan, were enrolled. In this period, more than 95% of the COVID-19 infections were reportedly caused by the Delta variant. As a result, a total of 784 adults with recent contact history, including 231 (29.5%) RT-PCR test-positive cases, were enrolled. The test positivity rate was lower in individuals who had been vaccinated twice than in unvaccinated individuals (12.5% vs. 39.0%, p0.0001), with the risk ratio of 0.32 (95% confidence interval 0.23-0.46). The vaccine effectiveness was the highest between 7-90 days after the second vaccine dose. In conclusion, two doses of mRNA COVID-19 vaccines effectively suppressed transmission in Japan during the nationwide pandemic of the Delta variant, estimated to have prevented 50-80% of the infection.

Objective Coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), remains the world's largest public health concern in 2021. A history of close contact with infectious patients is a factor that predicts a positive SARS-CoV-2 test result. Meanwhile, the precise predictive value of symptoms suggestive of COVID-19 has not been fully elucidated. This study aimed to clarify the predictive and discriminatory value of each clinical symptom suggestive of COVID-19. Methods This study enrolled participants who were tested for SARS-CoV-2 by reverse transcription polymerase chain reaction using a nasopharyngeal swab between November 2020 and January 2021. All enrolled patients were evaluated for data regarding the presence and closeness of contact with infectious patients and comprehensive clinical features (i.e., fever, cough, dyspnea, fatigue, dysosmia, and dysgeusia). Results Among the 1,744 tested participants, 144 tested positive for SARS-CoV-2. In the test-positive group, self-reported cough, fatigue, dysosmia, and dysgeusia were significant predictors of COVID-19, independent from a history of close contact. In particular, the presence of dysosmia was the strongest predictor of COVID-19 in both univariate and multivariate analyses. Among the 42 patients with self-reported dysosmia, 25 (59.5%) were SARS-CoV-2 test-positive. Self-reported dysosmia was reported by 25 (17.4%) of the 144 patients who tested positive for SARS-CoV-2, and 15 (60.0%) of the 25 COVID-19 patients with dysosmia had accompanying dysgeusia. Conclusion The presence of dysosmia was reported by 10-25% of patients with COVID-19, and is a significant predictor of COVID-19 infection, independent from a history of close contact.

Tombusviruses have been identified in several crops, including gentian virus A (GeVA) in Japanese gentian. In this study, we isolated another tombusvirus, Sikte waterborne virus strain C1 (SWBV-C1), from Japanese gentian. Although SWBV-C1 and GeVA are not closely related, SWBV-C1, like GeVA, showed host-specific low-temperature-dependent replication in gentian and arabidopsis. The use of in vitro transcripts from full-length cDNA clones of SWBV-C1 genomic RNA as inocula confirmed these properties, indicating that the identified genomic RNA sequences encode viral factors responsible for the characteristic features of SWBV-C1.

The exact profiles of the clinical symptoms related to the SARS-CoV-2 Omicron variant (B.1.1.529) remain largely uncertain. Therefore, this study aimed to clarify the clinical manifestations of infection with this variant. We enrolled individuals who were tested by quantitative nasopharyngeal swab reverse transcription-polymerase chain reaction (RT-PCR) test at a large screening center in a city of Japan during the B.1.1.529 Omicron variant wave between January and May 2022, after contact with COVID-19 patients. Swab tests were planned to be performed approximately 4-5 days after contact. The presence of COVID-19-related symptoms was assessed at the swab test site. Among the 2,507 enrolled individuals, 943 (37.6%) were RT-PCR test-positive and 1,564 (62.4%) were test-negative. Among the 943 PCR test-positive participants, the prevalence of the symptoms was as follows: 47.3% with cough, 32.9% with sore throat, 18.4% with fatigability, 12.7% with fever of ≥ 37.5℃, 9.9% with dyspnea, 2.1% with dysosmia, and 1.4% with dysgeusia. The prevalence of cough, sore throat, dyspnea, and fatigability was higher among adults aged ≥ 18 years than among children and adolescents. The prevalence of dysosmia and dysgeusia remarkably decreased during the Omicron wave (1-3%) compared to during the pre-Omicron variant waves (15-25%). In summary, common COVID-19-related symptoms during the Omicron variant wave included cough and sore throat, followed by fatigability, fever, and dyspnea. The prevalence of most of these symptoms was higher in adults than in non-adults. The prevalence of dysosmia and dysgeusia remarkably decreased with the Omicron variant than with pre-Omicron variants.

Antenatal depression (AD) is the commonest morbidity during pregnancy. There is evidence that premenstrual syndrome (PMS) and AD share common immune-inflammatory and sex hormonal pathways. This study aims to evaluate the association between the severity of depressive PMS and AD in early and late pregnancy.Participants were followed from early (=16 weeks) to late pregnancy (=20 weeks). The Premenstrual Symptoms Screening Tool (PSST) was used to assess PMS and AD symptoms were assessed using the Edinburgh Postnatal Depression Scale (EPDS).Up to 57.6% of the variance in the early EPDS score was explained by the regression on the first factor extracted from 10 depression and anxiety PSST items (dubbed the DepAnx PSST), insomnia PSST, relation dissatisfaction, and partner abuse. There were specific indirect effects of DepAnx PSST (The affective, but not psychosomatic, symptoms of PMS strongly predict AD symptoms suggesting that the pathophysiology of affective PMS symptoms overlap with those of AD.

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is the world's largest public health concern in 2021. This study evaluated the associations of the prevalence of airway symptoms among the tested individuals and data regarding the natural environmental factors with the weekly number of newly diagnosed COVID-19 patients in Sendai City (N

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), remains a global public health concern in 2021. However, the risk of attending schools during the pandemic remains unevaluated. This study estimated the secondary transmission rate at schools using the results of a real-time reverse transcription-polymerase chain reaction (RT-PCR) screening test performed between July 2020 and April 2021, before starting the nationwide mass vaccination. A total of 1,924 students (20 RT-PCR-positive; 1.0%) from 52 schools or preschools were evaluated, together with 1,379 non-adults (95 RT-PCR-positive; 6.9%) exposed to SARS-CoV-2 in non-school environments. Assuming that the infectious index cases were asymptomatic and the transmission at schools followed a Bernoulli process, we estimated the probability of transmission after each contact at school as approximately 0.005 (0.5% per contact) with the current infection prevention measures at schools in Japan (i.e., hand hygiene, physical distancing, wearing masks, and effective ventilation). Furthermore, assuming that all children are capable of carrying the infection, then contact between an index case and 20-30 students per day at schools would yield the expected value for secondary cases of ≥ 1.0, during the 10 days of the infectious period. In conclusion, with the current infection prevention measures at schools in Japan, secondary transmission at schools would occur in approximately every 200 contacts. When considering this rate, compliance with the current infection prevention measures at schools and early detection and quarantine of the index cases would be effective in preventing the spread of COVID-19 at schools.

Background Few studies examined the contributions of childhood adversities, intimate partner violence and social support to antenatal depression (AD). This study aims to 1) evaluate association of these psychosocial factors with AD symptoms in pregnancy; and 2) examine the mediating effect of social support on the relationship between psychosocial stressors and AD symptoms. Methods Participants were 120 pregnant women aged from 18 to 49 in less than 16 gestational weeks and attending at Antenatal Care Center at Khon Kaen hospital, Thailand. AD symptoms were assessed by the Edinburgh Postnatal Depression Scale (EPDS). Childhood adversities, intimate partner violence and social support were measured using the Adverse Childhood Experiences Questionnaire (ACE questionnaire), Abuse Assessment Screen (AAS), and Multidimensional Scale of Perceived Social Support (MSPSS). Results We found that the EPDS score was significantly and positively associated with adverse childhood experiences (ACEs) and negatively with social support. Partial Least Square analysis showed that 49.1% of the variance in the depressive subdomain of the EPDS score was predicted by ACEs, namely psychological and physical abuse and neglect, emotional or physical abuse by the partner, unplanned pregnancy, and no satisfaction with their relationship. The effects of adverse childhood experience due to neglect on the EDPS score was mediated by social support by friends. Limitations ACEs were assessed retrospectively and, therefore, may be susceptible to recall bias. Conclusion Prenatal depression scores are to a large extent predicted by psychological distress as indicated by early lifetime trauma, abuse by partner, relation satisfaction, and implications of unintended pregnancy.

Tombusviruses have been identified in several crops, which include gentian virus A (GeVA), in Japanese gentians. In this study, we isolated another tombusvirus, Sikte waterborne virus strain C1 (SWBV-C1) from Japanese gentian. Although SWBV-C1 and GeVA are not closely related among tombusviruses, SWBV-C1, like GeVA, showed host-specific low-temperature-preferred multiplication in gentians and Arabidopsis. The use of in vitro transcripts from full-length cDNA clones of SWBV-C1 genomic RNA as inocula confirmed these properties, which indicates that the identified genomic RNA sequences encode viral factors underlying characteristic SWBV-C1 features.

Tombusviruses have been identified in several crops, including gentian virus A (GeVA) in Japanese gentian. In this study, we isolated another tombusvirus, Sikte waterborne virus strain C1 (SWBV-C1), from Japanese gentian. Although SWBV-C1 and GeVA are not closely related, SWBV-C1, like GeVA, showed host-specific low-temperature-dependent replication in gentian and arabidopsis. The use of in vitro transcripts from full-length cDNA clones of SWBV-C1 genomic RNA as inocula confirmed these properties, indicating that the identified genomic RNA sequences encode viral factors responsible for the characteristic features of SWBV-C1.

In response to the COVID-19 pandemic caused by SARS-CoV-2 in 2020, we conducted drive-through nasopharyngeal swab testing for COVID-19 in Sendai city, Japan, since April 2020. All tested individuals were judged in advance by public health centers for the necessity of undergoing the test with possible contact history and/or symptoms suggestive of COVID-19. In this study, to identify the predictors of SARS-CoV-2 test positivity for more efficient and evidenced selection of suspected individuals, we enrolled 3,540 consecutive individuals, tested in the first 7 months of the testing program, with data regarding to the history of close contact with COVID-19 patients, including those involved in cluster outbreaks. This cohort included 284 foreign students (257 males and 27 females) from a vocational school involved in the largest cluster outbreak in the area. Close contact history was present in 952 (26.9%) of the participants. The reverse transcription-polymerase chain reaction (RT-PCR) test results showed that 164 participants (4.6%) were positive and 3,376 participants (95.4%) were negative for the SARS-CoV-2 nucleocapsid gene (N2). In the univariate and multivariate analyses, history of close contact with COVID-19 patients, higher age, cough symptoms, and non-native ethnicity were predictors for SARS-CoV-2 test positivity. However, the significance of age and foreign nationality disappeared or declined upon excluding the foreign students from the aforementioned largest cluster outbreak. In conclusion, a history of close contact with COVID-19 patients and the presence of cough symptoms are significant predictors of SARS-CoV-2 test positivity.

Clinical use of human embryonic stem cells (ESCs) as a raw material requires good manufacturing practice-compliant axillary materials such as culture medium. To this end, animal components should not be used and contamination of virus/bacteria/fungus and allergens are a concern. In addition, animal components such as albumin and fetal bovine serum pose difficulties such as a lot-to-lot variation. However, only a limited number of animal component-free media have been developed to date. In this study, we investigated whether SEES2 ESCs can be stably propagated for 16 passages (54 population doublings) over a period of 60 days in a newly established Stem-Partner ACF medium. SEES2 ESC maintained their intact karyotype, i.e. 46,XX, and their undifferentiated phenotypes after long-term culture. An in vitro differentiation assay revealed that SEES2 ESCs exhibited multipotency, i.e. endodermal, mesodermal and ectodermal differentiation. Subcutaneous implantation of SEES2 ESCs generated mature teratomas without malignant transformation. These results show that SEES2 ESCs in the Stem-Partner ACF medium can be used to establish master cell banks for future regenerative medicine as well as other ESCs in the previously reported culture medium.

During a disaster, all hospitals are expected to function as "social critical institutions" that protect the lives and health of people. In recent disasters, numerous hospitals were damaged, and this hampered the recovery of the affected communities. Had these hospitals business continuity plans (BCPs) to recover quickly after the disaster, most of the damage could have been avoided. This study conducted a scoping review of the historical trend and regional differences in hospital BCPs to validate the improvement of the BCP concept based on our own experience at Tohoku University Hospital, which was affected by the 2011 Great East Japan Earthquake and Tsunami (GEJET). We searched PubMed by using keywords related to BCP and adapted 97 articles for our analysis. The number of articles on hospital BCPs has increased in the 2000s, especially after Hurricane Katrina in 2005. While there are regional specificity of hazards, there were many common topics and visions for BCP implementation, education, and drills. From our 2011 GEJET experience, we found that BCPs assuming region-specific disasters are applicable in various types of disasters. Thus, we suggest the following integral and universal components for hospital BCPs: (1) alternative methods and resources, (2) priority of operation, and (3) resource management. Even if the type and extent of disasters vary, the development of BCPs and business continuity management strategies that utilize the abovementioned integral components can help a hospital survive disasters in the future.

The DECS (dsRNA isolation, exhaustive amplification, cloning and sequencing) analysis technique for viral diagnosis detected a tombusvirus in Japanese gentian not displaying severe symptoms. We tentatively named this virus "gentian virus A" (GeVA). GeVA systemically but inefficiently infected Japanese gentian without causing visible symptoms, while it led to severe symptoms in some other plants. The complete genome sequence of GeVA indicated a typical tombusvirus-like structure. Phylogenetic analysis of the deduced amino acid sequences of four tombusvirus-encoded proteins did not reveal other known tombusviruses that were closely-related to GeVA, suggesting that it is a novel tombusvirus.

Gentian virus A (GeVA), a novel tombusvirus isolated from Japanese gentian, has shown only a limited ability to infect Japanese gentians under experimental conditions. In this study, temperature was found to affect the efficient multiplication of GeVA in Japanese gentians. GeVA efficiently multiplied in inoculated leaves of gentians at 18 °C but not at 23 °C. This low-temperature (18 °C)-preferred GeVA multiplication was specifically observed in Japanese gentians and Arabidopsis thaliana but not in other experimental plants, including Nicotiana benthamiana. In A. thaliana, visible defense responses, including pathogenesis-related protein 1 expression, were not detected at 23 °C. Furthermore, several A. thaliana mutants, including those defective in RNA silencing, with altered plant immunities did not allow GeVA to multiply to detectable levels at 23 °C. Taken together, these data suggest that unique interaction between GeVA and gentians/A. thaliana, which is independent of RNA silencing, may underlie the low-temperature-preferred multiplication of GeVA.

Plant stomata represent the main battlefield for host plants and the pathogens that enter plant tissues via stomata. Septoria spp., a group of ascomycete fungi, use host plant stomata for invasion and cause serious damage to agricultural plants. There is no evidence, however, showing the involvement of stomata in defense systems against Septoria infection. In this study, we isolated Septoria gentianae 20-35 (Sg20-35) from Gentiana triflora showing gentian leaf blight disease symptoms in the field. Establishment of an infection system using gentian plants cultured in vitro enabled us to observe the Sg20-35 infection process and estimate its virulence in several gentian cultivars or lines. Sg20-35 also entered gentian tissues via stomata and showed increased virulence in G. triflora compared with G. scabra and their interspecific hybrid. Notably, the susceptibility of gentian cultivars to Sg20-35 was associated with their stomatal density on the adaxial but not abaxial leaf surface. Treatment of EPIDERMAL PATTERNING FACTOR-LIKE 9 (EPFL9/STOMAGEN) peptides, a small secreted peptide controlling stomatal density in Arabidopsis thaliana, increased stomatal density on the adaxial side of gentian leaves as well. Consequently, treated plants showed enhanced susceptibility to Sg20-35. These results indicate that stomatal density on the adaxial leaf surface is one of the major factors determining the susceptibility of gentian cultivars to S. gentianae and suggest that stomatal density control may represent an effective strategy to confer Septoria resistance.

A 1-year-old female toddler who was under a great amount of psychological stress presented with acute hematemesis. She had no significant medical history; she displayed lethargy and was anemic. Although blood transfusion and famotidine were administered for upper gastrointestinal bleeding (UGIB), the anemia did not improve. Thus, early endoscopy was performed under general anesthesia, revealing a gastric ulcer with exposed vessels in the supra-angular region. Cauterization via bipolar hemostatic forceps was performed on the exposed vessels, and the bleeding was controlled successfully. We diagnosed the patient with hemorrhagic stress-induced gastric ulcer due to psychological stress. Three months later, endoscopy revealed that the gastric ulcer had healed.

Vesicle trafficking including the exocytosis pathway is intimately associated with host immunity against pathogens. However, we still have insufficient knowledge about how it contributes to immunity, and how pathogen factors affect it. In this study, we explore host factors that interact with the Magnaporthe oryzae effector AVR-Pii. Gel filtration chromatography and co-immunoprecipitation assays identified a 150 kDa complex of proteins in the soluble fraction comprising AVR-Pii and OsExo70-F2 and OsExo70-F3, two rice Exo70 proteins presumably involved in exocytosis. Simultaneous knockdown of OsExo70-F2 and F3 totally abrogated Pii immune receptor-dependent resistance, but had no effect on Pia- and Pik-dependent resistance. Knockdown levels of OsExo70-F3 but not OsExo70-F2 correlated with reduction of Pii function, suggesting that OsExo70-F3 is specifically involved in Pii-dependent resistance. Under our current experimental conditions, over-expression of AVR-Pii or knockdown of OsExo70-F2 and -F3 genes in rice did not affect the virulence of compatible isolates of M. oryzae. AVR-Pii interaction with OsExo70-F3 appears to play a crucial role in immunity triggered by Pii, suggesting a role for OsExo70 as a decoy or helper in Pii/AVR-Pii interactions. Significance Statement In monocots, evidence for accessory proteins in effector-triggered immunity (ETI) is limited. We show that recognition of the Magnaporthe oryzae avirulence effector AVR-Pii mediated by the resistance (R) protein Pii is not direct, but requires an exocyst protein, Exo70, supporting Exo70 as either a "decoy" or a "helper".

The silk industry, which was among the leaders of modern industry in Japan, was formerly a mainstayof the Japanese economy. It flourished from the last days of the Edo period through the Meiji Restoration,an age when modern culture flowed into Japan from the West. The Tomioka silk mill in Gunma wasestablished in 1872 under government management, and the silk industry spread out from there to everypart of Japan.Silk was a familiar material, used for kimono and other kinds of clothes and bedding. When syntheticfibers came on the market, silk left the world of the kimono, and in recent years a silk kimono is almostnever worn. Silk has a luster which isn't seen in other material; in lightness, feel, quality and the way thefabric breathes, it is a remarkably excellent material. The charm of silk, which comes from its beauty andthe way the fabric breathes, cannot be found in other fibers. It is really the queen of fibers. I would like topropose that this wonderful material, a world treasure of which Gunma can be proud, can be put to creativeuse in the fashion business field.

Prader-Willi syndrome (PWS), a complex genetic disorder, arises from suppressed expression of paternally inherited imprinted genes on chromosome 15q11-q13. Characteristics include short stature, intellectual disability, behavioral problems, hypogonadism, obesity, and reduced bone and muscle mass. Testosterone replacement (TR) remains controversial due to concerns regarding behavioral problems. To evaluate the effects of TR on secondary sexual characteristics, body composition, and behavior in adult males with PWS, 22 male PWS patients over the age of 16 with behavioral scores of less than grade 4 on the Modified Overt Aggression Scale (MOAS) underwent monthly intramuscular TR (125 mg). Pubertal change, body composition and behavior were evaluated before and after 24 months of therapy. Serum testosterone, LH, and FSH did not change. Increased pubic hair was observed in 16 of 22 patients (72.7%). Percent body fat decreased from 47.55 ± 2.06% to 39.75 ± 1.60% (n = 18) (P = 0.018). Bone mineral density increased from 0.8505 ± 0.0426 g/cm(2) to 0.9035 ± 0.0465 g/cm(2) (n = 18) (P = 0.036), and lean body mass increased from 18093.4 ± 863.0 g to 20312.1 ± 1027.2 g (n = 18) (P = 0.009). The MOAS was unchanged, from 4.5 ± 2.0 at the beginning of the study to 3.0 ± 1.7 at the end of study indicating no increase in aggression. No behavioral problems were observed. Based on this pilot study, TR with 125 mg monthly is a potentially safe and useful intervention for adult males with PWS.

A 9.0-magnitude earthquake occurred off the northeast coast of Japan at 2:46 p.m. (local time) on Friday, 11 March 2011, with the epicenter approximately 70 km (43 mi) east of the Oshika Peninsula and the hypocenter at a depth of approximately 32 km (20 mi) below sea level [Figure 1(a)] [1]. The earthquake triggered powerful tsunami waves, which reached heights of up to 40.5 m (133 ft) in Miyako and traveled around 4 km (about 2.5 mi) inland in the Sendai area. As of 30 March 2012, a total of 18,897 people were either dead or missing.

IntroductionThere were 5,385 deceased and 710 missing in the Ishinomaki medical zone following the Great East Japan Earthquake that occurred in Japan on March 11, 2011. The Ishinomaki Zone Joint Relief Team (IZJRT) was formed to unify the relief teams of all organizations joining in support of the Ishinomaki area. The IZJRT expanded relief activity as they continued to manually collect and analyze assessments of essential information for maintaining health in all 328 shelters using a paper-type survey. However, the IZJRT spent an enormous amount of time and effort entering and analyzing these data because the work was vastly complex. Therefore, an assessment system must be developed that can tabulate shelter assessment data correctly and efficiently. The objective of this report was to describe the development and verification of a system to rapidly assess evacuation centers in preparation for the next major disaster.ReportBased on experiences with the complex work during the disaster, software called the “Rapid Assessment System of Evacuation Center Condition featuring Gonryo and Miyagi” (RASECC-GM) was developed to enter, tabulate, and manage the shelter assessment data. Further, a verification test was conducted during a large-scale Self-Defense Force (SDF) training exercise to confirm its feasibility, usability, and accuracy. The RASECC-GM comprises three screens: (1) the “Data Entry screen,” allowing for quick entry on tablet devices of 19 assessment items, including shelter administrator, living and sanitary conditions, and a tally of the injured and sick; (2) the “Relief Team/Shelter Management screen,” for registering information on relief teams and shelters; and (3) the “Data Tabulation screen,” which allows tabulation of the data entered for each shelter, as well as viewing and sorting from a disaster headquarters’ computer. During the verification test, data of mock shelters entered online were tabulated quickly and accurately on a mock disaster headquarters’ computer. Likewise, data entered offline also were tabulated quickly on the mock disaster headquarters’ computer when the tablet device was moved into an online environment.ConclusionsThe RASECC-GM, a system for rapidly assessing the condition of evacuation centers, was developed. Tests verify that users of the system would be able to easily, quickly, and accurately assess vast quantities of data from multiple shelters in a major disaster and immediately manage the inputted data at the disaster headquarters.IshiiT, NakayamaM, AbeM, TakayamaS, KameiT, AbeY, YamaderaJ, AmitoK, MorinoK. Development and verification of a mobile shelter assessment system “Rapid Assessment System of Evacuation Center Condition featuring Gonryo and Miyagi (RASECC-GM)” for major disasters. Prehosp Disaster Med. 2016;31(5):539–546.

Background: Recently, the role of aldosterone in metabolic syndrome (MS) has aroused interest and several reports have suggested that aldosterone blockade could be beneficial in reducing blood pressure (BP). Methods: To examine the add-on effects of eplerenone (EP) on BP in patients with MS, 54 hypertensive patients with MS and 44 without MS were recruited. Systolic and diastolic BPs in mmHg before the initiation of EP was 144/84 ± 13/12 (MS group) and 147/85 ± 12/14 (non-MS group). Before the start of EP, all patients in both groups were treated with at least one antihypertensive drug. BPs were checked on every visit (at least every 2 months) and serum chemistries were measured every 4 months. The levels of microalbuminuria and aminoterminal pro-brain natriuretic peptide (NT pro-BNP) were determined before the start of and at the end of the study. Patients were followed for 1 year. If adverse effects were reported by patients or found in laboratory studies, EP was withdrawn. Results: One month after the start of EP, BPs were decreased to 140/80 ± 12/12 mmHg (MS group) versus 142/82 ± 11/12 mmHg (non-MS group) and there was no difference between the two groups. Towards the end of the study, BPs of both groups gradually decreased. At the end of the study, BPs of both groups were 129/76 ± 15/13 mmHg (MS group) versus 133/78 ± 13/11 mmHg (non-MS group). There was a significant difference in reduction of systolic BP between the two groups (p < 0.05). Add-on EP significantly decreased the levels of urinary excretion of albumin in MS patients but not in non-MS patients (p < 0.05). There was a significant correlation between reduction of systolic BP and NT pro-BNP but not microalbuminuria in the MS group (p < 0.05). There were no serious adverse effects in both groups. Conclusion: EP may have some beneficial effects in lowering BP in patients with reduction of microalbuminuria.

Growth hormone (GH) therapy is now widely given to Prader-Willi syndrome (PWS) patients to encourage growth in body height and to prevent obesity. Scoliosis, one of the complications in this syndrome, is thought to be accelerated in parallel with a rapid increase in body height, especially during adolescence. To determine whether GH therapy aggravates scoliosis and to identify any factor which might predict the progression of scoliosis, we studied 35 (22 males and 13 female) PWS patients between the ages of 2-16 years on GH therapy whose scoliosis was followed with spinal X-rays every 6 months. Thirteen (37.1%) of 35 patients had scoliosis with a Cobb angle of over 10°. Scoliosis was unchanged in five patients (14.3%), became worse in six (17.1%) and improved in two (5.7%). All 22 (62.9%) of 35 patients who did not have scoliosis did not develop scoliosis with GH therapy. Since abnormal paraspinal muscle development was thought to induce scoliosis, we measured cross-sectional areas of paraspinal muscles by using one slice CT scan at the level of the umbilicus at the level of L4. Since there was a delay in the increase in total paravertebral muscle area and prolonged asymmetry in patients with progressive scoliosis, both were thought to be useful predictors of progressive scoliosis in PWS patients with GH therapy.

In this study, the processes responsible for an intense wind event that occurred in west Sumatera on November 19, 2006, during the first campaign of the Hydrometeorological ARay for Isv-Monsoon AUtomonitoring (HARIMAU) were investigated. Strong winds of 17 m s^[-1] and a sudden temperature drop of 5 K were observed at an X-band Doppler radar site associated with the passage of a convective system, and some houses were severely damaged. The convective system developed under an environment of strong low-level easterly vertical shear associated with the easterly region of an equatorial Rossby wave. The northern part of the convective system possessed qualitatively similar structures to midlatitude bow echoes, including the convex shape of the convective line, a descending rear-inflow jet positioned at its apex, and mesoscale vortices on both sides of the rear-inflow jet. The low-level wind behind the convective system formed a channel of strong easterly wind as it passed through an area of relatively low topography in the mountain range. The enhanced easterly wind was thought to contribute to the formation of the bow echo-like structure in the northern part of the convective system. This easterly rear-inflow jet was further accelerated in the convective system and descended near the leading edge, forming divergent strong winds at the surface. The sounding data that were taken after the passage of the convective system indicated that dry air appeared in the lower troposphere associated with an enhancement of the southerly component of the wind. An analysis of objective reanalysis data suggests that the southerly was probably associated with westward-propagating mixed Rossby-gravity waves with a period of approximately 5 days. It is suggested that the dry air intruded into the convective system across the back edge of the precipitation area and caused enhanced evaporative cooling, which resulted in the effective downward transport of the enhanced easterly momentum.

West症候群の第一選択治療はACTH療法 (AT) であるが，腎石灰化，腎結石合併の報告もある。本症で併用されることが多いzonisamide (ZNS) は，炭酸脱水素酵素阻害剤様の作用を持ち，腎結石の有害事象がある。 AT中，腎石灰化を認めた5例についてZNS投与の有無，AT期間，ACTH総量，AT中の尿中Ca/Cr比，%TRP，尿pH，血清Ca，P，ALP値の推移を後方視的に検討し，腎石灰化を起こさなかった4例と比較した。腎石灰化を認めた全例でZNSを併用していた。AT期間は腎石灰化群で長く量も多い傾向があった。AT中の尿中Ca/Cr比の上昇は，腎石灰化群でより高かった。血清P，%TRPは低下し，血清ALPも低下したが両群で差はなかった。ZNSの併用，長期多量のAT，尿中Ca/Cr比の上昇が腎石灰化の危険因子と思われた。AT中，特にZNS併用時は，腎エコー，尿中Ca/Cr比による経過観察が必要である。

Ornamental gentian plants are perennial and have a juvenile period of over 1 year before flowering. We transformed gentian plants with a construct comprising the ArabidopsisFLOWERING LOCUS T (FT) gene (encoding a major component protein of the flowering hormone ‘florigen’) under the control of the rolC promoter from Agrobacterium rhizogenes, which is known to induce vascular-specific expression. The resultant rolCpro-FT transgenic gentian plants showed early flowering in vitro and the earliest line formed floral buds within 4 months after transformation. Regeneration experiments from leaf explants of these rolCpro-FT transgenic plants also confirmed the early flowering phenotype. After acclimatization, these transgenic plants showed normal floral development in a closed greenhouse. There is no effective method to induce early flowering by cultivation management in gentian, therefore these lines might be very useful as annual early season cultivars.

Gentian is one of the most important ornamental flowers in Japan. Gentiana triflora , G. scabra and their interspecific hybrids are the main breeding materials. Gentian cultivars are easily proliferated vegetatively, therefore it is important to develop a reliable discrimination method to prevent the illegal propagation and distribution of various high-value cultivars. Here, we report five sequence characterized amplified region (SCAR) markers based on the length polymorphisms in introns of four gentian flavonoid biosynthetic genes. These SCAR markers effectively discriminated nine gentian cultivars and nine breeding lines. This method could be applied in identifying gentian cultivars/lines and therefore will aid in protecting breeders’ rights.

RNA interference (RNAi) is an efficient and powerful technique for gene silencing compared with antisense and sense suppression. Here we report adaptation of RNAi technology to modify flower colors in gentian, targeted for suppression of three anthocyanin biosynthetic genes; chalcone synthase (CHS), anthocyanidin synthase (ANS) and flavonoid 3′,5′-hydroxylase (F3′5′H). The petals of transgenic gentian plants with a suppressed CHS gene exhibited pure white to pale-blue color, while those with a suppressed ANS gene showed only pale-blue. The suppression of the F3′5′H gene decreased delphinidin derivatives and increased cyanidin derivatives, and led to magenta flower colors. Northern blot analyses confirmed that all transgenic gentian plants showing typical phenotypes had strongly suppressed transcriptions of the targeted genes, corresponding with a change in anthocyanin accumulation and composition in their petals. Some rolCpro-CHSir transgenic gentians exhibited bicolor phenotypes with reduced anthocyanin accumulation along the vascular bundles. These data demonstrated that the suppression of anthocyanin biosynthetic genes by RNAi was successfully applied to gentian plants to change flower color, and this could be useful for designing novel flower color and patterns. Transgenic gentian plants produced in this study might be utilized as elite materials in the breeding of gentian plants in the near future.

Orange- to red-colored flowers are difficult to produce by conventional breeding techniques in some floricultural plants. This is due to the deficiency in the formation of pelargonidin, which confers orange to red colors, in their flowers. Previous researchers have reported that brick-red colored flowers can be produced by introducing a foreign dihydroflavonol 4-reductase (DFR) with different substrate specificity in Petunia hybrida, which does not accumulate pelargonidin pigments naturally. However, because these experiments used dihydrokaempferol (DHK)-accumulated mutants as transformation hosts, this strategy cannot be applied directly to other floricultural plants. Thus in this study, we attempted to produce red-flowered plants by suppressing two endogenous genes and expressing one foreign gene using tobacco as a model plant. We used a chimeric RNAi construct for suppression of two genes (flavonol synthase [FLS] and flavonoid 3'-hydroxylase [F3'H]) and expression of the gerbera DFR gene in order to accumulate pelargonidin pigments in tobacco flowers. We successfully produced red-flowered tobacco plants containing high amounts of additional pelargonidin as confirmed by HPLC analysis. The flavonol content was reduced in the transgenic plants as expected, although complete inhibition was not achieved. Expression analysis also showed that reduction of the two-targeted genes and expression of the foreign gene occurred simultaneously. These results demonstrate that flower color modification can be achieved by multiple gene regulation without use of mutants if the vector constructs are designed resourcefully.

The immunotoxicity of anticancer drugs has been measured with whole blood TNF-alpha production induced by LPS-stimulation. Cis-platinum (CDDP) or 5-fluorouracil (5-FU) was given intravenously to mice once a day for three days. The amount of produced-TNF-alpha lowered significantly in CDDP 2 mg/kg and 5 mg/kg, while the number of leukocytes scarcely changed even when a significant decrease in body weight was observed at the dose of 5 mg/kg. On the other hand, 5-FU 20 mg/kg also caused a significant decline in the amount of produced TNF-alpha but it showed no apparent effects on the number of leukocytes and the body weight. These results indicate that LPS-induced TNF-alpha production is a more sensitive measure than leukocyte count used conventionally to detect the immunotoxicity of two anticancer drugs, CDDP and 5-FU, and suggest that it might be a more practical index for hazard identification in clinical treatment with anticancer drugs.

We successfully produced dwarf potted gentian using wild-type Agrobacterium rhizogenes A4 strain (ATCC43057) harboring an agropine type plasmid (pRiA4). A number of hairy roots were induced by direct inoculation with A. rhizogenes to the stem and leaf tissues in vitro. Adventitious shoots were regenerated from the hairy roots on Murashige and Skoog medium containing 10 mg l � 1 N-phenyl-N� -(1,2,3-thiadiazol-5-yl) urea, 0.1 or 1.0 mg l � 1 1-naphthaleneacetic acid, and 3 g l � 1 gellan gum. Regenerated plants were then cultured in vitro for at least six months and tested for the absence of viable A. rhizogenes within their tissues. After receiving authorization from the Ministry of Agriculture, Forestry and Fisheries, Japan, the plants were acclimated and grown in a greenhouse and/or field. Opine synthesis and rolC gene expression were analyzed to demonstrate successful introduction and expression of the transferred DNA. In total, 122 lines of variant types of dwarf plants with blue, white, and pink flowers were obtained, which might be useful for molecular breeding of a series of dwarf potted gentian cultivars.

To produce white-flowered gentian plants, we attempted to suppress the chalcone synthase (chs) gene by Agrobacterium-mediated transformation. A binary vector, pSMABcCHS, harboring antisense cDNA of chs isolated from the gentian Gentiana triflora cv. Maciry under control of the CaMV35S promoter was transformed into an interspecific hybrid gentian (cv. Albireo; G. scabraG. triflora). The vector also contained the bar gene as a selectable marker. Three out of seventeen transgenic plants showed completely white flowers with 10 to 25% reduced anthocyanin content compared with the wild-type. Molecular analyses confirmed integration of the foreign genes and suppression of chs mRNA accumulation in their petals. Application of commercial herbicide including bialaphos showed that the transformants were strongly resistant. Segregation of the white-flowered and herbicide-resistant traits was tested using a T1 progeny obtained from crossing with a blue-flowered parental line. The results clearly showed that these two traits are inherited dominantly as linked traits in the T1 progeny, suggesting that these transgenic plants are useful resources for production of white-flowered gentians. These results also demonstrated for the first time the inheritance of foreign genes and genetic modification of flower color in transgenic gentian plants.

Summary In this study, no transgenic gentian (Gentiana triflora × Gentiana scabra) plants produced via Agrobacterium-mediated transformation exhibited transgene (GtMADS, gentian-derived MADS-box genes or sGFP, green fluorescent protein) expression in their leaf tissues, despite the use of constitutive Cauliflower mosaic virus (CaMV) 35S promoter. Strikingly, no expression of the selectable marker gene (bar) used for bialaphos selection was observed. To investigate the possible cause of this drastic transgene silencing, methylation-specific sequences were analysed by bisulfite genomic sequencing using tobacco transformants as a control. Highly methylated cytosine residues of CpG and CpWpG (W contains A or T) sites were distinctively detected in the promoter and 5′ coding regions of the transgenes 35S-bar and 35S-GtMADS in all gentian lines analysed. These lines also exhibited various degrees of cytosine methylation in asymmetrical sequences. The methylation frequencies in the other transgene, nopaline synthase (NOS) promoter-driven nptII, and the endogenous GtMADS gene coding region, were much lower and were variable compared with those in the 35S promoter regions. Transgene methylation was observed in the bialaphos-selected transgenic calluses expressing the transgenes, and methylation sequences were distributed preferentially around the as-1 element in the 35S promoter. Calluses derived from leaf tissues of silenced transgenic gentian also exhibited transgene suppression, but expression was recovered by treatment with the methylation inhibitor 5-aza-2′-deoxycytidine (aza-dC). These results indicated that cytosine methylation occurs exclusively in the 35S promoter regions of the expressed transgenes during selection of gentian transformants, causing transcriptional gene silencing.

To quantify the nitrogen (N) use by Pinus densiflora trees growing on an infertile lava surface, N pools, N requirement and N uptake through fine roots and N deposition from the atmosphere were estimated. The N requirement and the N uptake of fine roots were 55.5 kg N ha−1 year−1 and 39.7 kg N ha−1, respectively. Thus, the ratio of N uptake to N requirement of the fine roots was 71.5%. Including fine-root contribution, the total N requirement of the P. densiflora trees was 98.6 kg N ha−1 year−1, and the total N uptake was 64.2 kg N ha−1 year−1. Thus, the N uptake of the P. densiflora trees was 64.1% of the N requirement, indicating that P. densiflora trees growing on an infertile lava surface obtain some of their N from below-ground organic material layers every year and the contribution of N storage in trees for their growth is not any higher than indicated in previous reports that excluded fine-roots contribution. The wet N deposition of our research forest was only 5.8% of the N requirement of the P. densiflora trees and only 8.9% of the N uptake. Movement of the below-ground organic material layer N concentrations in the F- and L-layers coincides with needle development and fine-root growth, suggesting the possibility that P. densiflora trees extract N from the organic N of those layers for growth.