Arabidopsis CURLY LEAF functions in leaf immunity against fungal pathogens by concomitantly repressing SEPALLATA3 and activating ORA59.

Arabidopsis non-host resistance against non-adapted fungal pathogens including Colletotrichum fungi consists of pre-invasive and post-invasive immune responses. Here we report that non-host resistance against non-adapted Colletotrichum spp. in Arabidopsis leaves requires CURLY LEAF (CLF), which is critical for leaf development, flowering and growth. Microscopic analysis of pathogen behavior revealed a requirement for CLF in both pre- and post-invasive non-host resistance. The loss of a functional SEPALLATA3 (SEP3) gene, ectopically expressed in clf mutant leaves, suppressed not only the defect of the clf plants in growth and leaf development but also a defect in non-host resistance against the non-adapted Colletotrichum tropicale. However, the ectopic overexpression of SEP3 in Arabidopsis wild-type leaves did not disrupt the non-host resistance. The expression of multiple plant defensin (PDF) genes that are involved in non-host resistance against C. tropicale was repressed in clf leaves. Moreover, the Octadecanoid-responsive Arabidopsis 59 (ORA59) gene, which is required for PDF expression, was also repressed in clf leaves. Notably, when SEP3 was overexpressed in the ora59 mutant background, C. tropicale produced clear lesions in the inoculated leaves, indicating an impairment in non-host resistance. Furthermore, ora59 plants overexpressing SEP3 exhibited a defect in leaf immunity to the adapted Colletotrichum higginsianum. Since the ora59 plants overexpressing SEP3 did not display obvious leaf curling or reduced growth, in contrast to the clf mutants, these results strongly suggest that concomitant SEP3 repression and ORA59 induction via CLF are required for Arabidopsis leaf immunity to Colletotrichum fungi, uncoupled from CLF's function in growth and leaf development.

[Validation Study on an Improved Quantitative Method for Aflatoxins in Foods].

As an analytical method for aflatoxins in foods, the analytical method based on the notification by the director of the Food Safety Department, Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare (August 16, 2011) has been established. In order to improve the operability and analytical performance of the conventional method, this study aimed to construct an improved method that optimized selection of immunoaffinity column (IAC) and purifying condition, and omitted evaporation after the purification with IAC. In the recovery test performed by adding 2.5 ng/g of aflatoxin B1, B2, G1 and G2 standard solutions into 9 kinds of food samples, the improved method achieved the established target values: 77.0-99.7% of recovery, 1.7-5.6% of intra-assay coefficient of validation, and 0.9-3.6% of inter-assay of coefficient of variation, respectively. The improved method also achieved 4.3-10.5% greater recovery and 1.5 hours shorter preparation time than the conventional one. These results indicate applicability of the improved method for 9 kinds of foods and its efficacy as an analytical method for aflatoxins in foods.

Blood Pressure Elevation of Tubular Specific (P)RR Transgenic Mice and Lethal Tubular Degeneration due to Possible Intracellular Interactions between (P)RR and Alternative Renin Products.

The prorenin/renin receptor ((P)RR) is a multifunctional protein that is widely distributed in various organs. Despite intensive research for more than 20 years, this receptor has not been fully characterized. In this study, we generated mice overexpressing the tubular epithelial (P)RR gene ((P)RR-TG mice) to test the previously reported functional role of (P)RR by Ramkumar et al. in 2015 using tubular specific (P)RR KO mice. (P)RR-TG mice were maintained and analyzed in individual metabolic cages and were administered angiotensin II blocker (ARB), direct renin inhibitor (DRI), and bafilomycin, that is, vacuolar ATPase (V-ATPase) antagonist. (P)RR-TG mice were hypertensive and had alkalized urine with lower osmolality and Na+ excretion. ARB and DRI, but not bafilomycin, concurrently decreased blood pressure. Bafilomycin acidized urine of (P)RR-TG mice, or equivalently this phenomenon restored the effect of overexpressed transgene, suggesting that (P)RR functioned as a V-ATPase in renal tubules. Afterall, (P)RR-TG mice were mated with alternative renin transgenic mice (ARen2-TG), which we identified as intracellular renin previously, to generate double transgenic mice (DT-TG). Lethal renal tubular damage was observed in DT-TG mice, suggesting that intracellular renin may be a ligand for (P)RR in tubules. In summary, (P)RR did not substantially affect the tissue renin-angiotensin system (RAS) in our model of tubular specific (P)RR gene over-expression, but alternative intracellular renin may be involved in (P)RR signaling in addition to conventional V-ATPase function. Further investigations are warranted.

[Development of an LC-MS/MS-Based Rapid and Simple Analytical Method for Six Fungicides].

In this study, we developed an LC-MS/MS-based rapid and simple analytical method for six fungicides; imazalil, o-phenylphenol, thiabendazole, fludioxonil, azoxystrobin and pyrimethanil, the latter three were newly approved for use after 2011. For expediting and simplification, we merged the extraction method with that of the pesticide analysis. For purification step, loading of 1 mL of sample extracts to 500 mg Oasis HLB column and elution with 8 mL of acetonitrile gave satisfactory results. The performance of the present method was confirmed for orange, grapefruit, and lemon samples fortified with the six fungicides. The results showed that the average recovery ranged from 89.7 to 100.0%, intra- and inter-assay CV% ranged from 1.5 to 5.0% and from 0.5 to 4.9%, respectively, achieving the target values of the Japanese official guideline for residual pesticide analysis. The limits of quantification of this method were determined to be 1 mg/kg for o-phenylphenol, and 0.2 mg/kg for the other five fungicides. These values were lower than their corresponding regulation values. In addition, we confirmed the usability of the present method for fungicide inspection of commercially available citrus fruits. During 2017-2019, there was no conflict between the food labeling and the fungicides detected and no fungicide with the concentration exceeding maximum residue level was detected.

Problems and solutions of polyethylene glycol co-injection method in multiresidue pesticide analysis by gas chromatography-mass spectrometry: evaluation of instability phenomenon in type II pyrethroids and its suppression by novel analyte protectants.

Polyethylene glycol 300 is commonly used as a base material for "analyte protection" in multiresidue pesticide analysis via gas chromatography-mass spectrometry. However, the disadvantage of the co-injection method using polyethylene glycol 300 is that it causes peak instability in α-cyano pyrethroids (type II pyrethroids) such as fluvalinate. In this study, we confirmed the instability phenomenon in type II pyrethroids and developed novel analyte protectants for acetone/n-hexane mixture solution to suppress the phenomenon. Our findings revealed that among the examined additive compounds, three lipophilic ascorbic acid derivatives, 3-O-ethyl-L-ascorbic acid, 6-O-palmitoyl-L-ascorbic acid, and 6-O-stearoyl-L-ascorbic acid, could effectively stabilize the type II pyrethroids in the presence of polyethylene glycol 300. A mixture of the three ascorbic acid derivatives and polyethylene glycol 300 proved to be an effective analyte protectant for multiresidue pesticide analysis. Further, we designed and evaluated a new combination of analyte protectant compounds without using polyethylene glycol or the troublesome hydrophilic compounds. Consequently, we obtained a set of 10 medium- and long-chain saturated fatty acids as an effective analyte protectant suitable for acetone/n-hexane solution that did not cause peak instability in type II pyrethroids. These analyte protectants will be useful in multiresidue pesticide analysis by gas chromatography-mass spectrometry in terms of ruggedness and reliable quantitativeness. Graphical abstract Comparison of effectiveness of the addition of lipophilic derivatives of ascorbic acid in controlling the instability phenomenon of fluvalinate with polyethylene glycol 300.

Detection of antibiotics in chicken eggs obtained from supermarkets in Ho Chi Minh City, Vietnam.

The residual levels of antibiotics in Vietnamese eggs were monitored from 2014 to 2015. A total of 111 egg packages, distributed by 11 different companies, were collected from supermarkets in Ho Chi Minh City and the levels of 28 antibiotics were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) screening method. Sixteen samples tested positive for antibiotics; a total of eight compounds (enrofloxacin, ciprofloxacin, norfloxacin, sulfadimethoxine, sulfamethazine, sulfamonomethoxine, tilmicosin and trimethoprim) were detected. Enrofloxacin was detected in eight samples, with two samples exhibiting concentrations exceeding 1,000 µg kg-1. Tilmicosin was detected in three samples at a range of 49-568 µg kg-1. We observed that two of the 11 companies frequently sold antibiotic-contaminated eggs (detection rates of 56 and 60%), suggesting that a number of companies do not regulate the use of antibiotics in egg-laying hens. Our findings indicate that livestock farmers require instruction regarding antibiotic use and that continual antibiotic monitoring is essential in Vietnam.

Enterococcus saigonensis sp. nov., isolated from retail chicken meat and liver.

Two Gram-stain-positive strains, VE80T and VE116, which were resistant to vancomycin, were isolated from retail chicken meat and liver in Ho Chi Minh, Vietnam, respectively. These strains were characterized by sequence analyses of 16S rRNA, RNA polymerase α-subunit (rpoA), ATP synthase α-subunit (atpA), and phenylalanyl-tRNA synthase α-subunit (pheS) genes, determination of DNA G+C content, cellular fatty acid methyl ester analysis, DNA-DNA hybridization, and conventional morphological and biochemical tests. Strains VE80T and VE116 had 99.6 % 16S rRNA gene sequence similarity with Enterococcus canintestini LMG 13590T, and 99.1 % 16S rRNA gene sequence similarity with Enterococcus dispar ATCC 51266T. However, the two isolates could be clearly differentiated from these reference strains by the low sequence similarities (86.1-86.8 %) of the atpA gene, low DNA-DNA relatedness (<22.8 %), and differences in the production of acid from melezitose and methyl α-d-glucoside. Based on the results obtained in the present study, these two isolates are considered to represent a novel species of the genus Enterococcus, for which the name Enterococcus saigonensis sp. nov., is proposed. The type strain is VE80T (=JCM 31193T=CCUG 68827T).

Evidence-based Effective Triage Operation During Disaster: Application of Human-trajectory Data to Triage Drill Sessions.

INTRODUCTION: Though many governmental and nongovernmental efforts for disaster prevention have been sought throughout Japan since the Great East Japan Earthquake on March 11, 2011, most of the preparation efforts for disasters have been based more on structural and conventionalized regulations than on scientific and objective grounds. Problem There has been a lack of scientific knowledge for space utilization for triage posts in disaster drill sessions. This report addresses how participants occupy and make use of the space within a triage post in terms of areas of use and occupied time. METHOD: The trajectories of human movement by using Ubiquitous Stereo Vision (USV) cameras during two emergency drill sessions held in 2012 in a large commercial building have been measured. The USV cameras collect each participant's travel distance and the wait time before, during, and after undergoing triage. The correlation between the wait time and the space utilization of patients at a triage post has been analyzed. RESULTS: In the first session, there were some spaces not entirely used. This was caused largely by a patient who arrived earlier than others and lingered in the middle area, which caused the later arrivals to crowd the entrance area. On the other hand, in the second session, the area was used in a more evenly-distributed manner. This is mainly because the earlier arrivals were guided to the back space of the triage post (ie, the opposite side of the entrance), and the late arrivals were also guided to the front half, which was not occupied by anyone. As a result, the entire space was effectively utilized without crowding the entrance. CONCLUSION: This study has shown that this system could measure people's arrival times and the speed of their movements at the triage post, as well as where they are placed until they receive triage. Space utilization can be improved by efficiently planning and controlling the positioning of arriving patients. Based on the results, it has been suggested that for triage operation, it is necessary to efficiently plan and control the placement of patients in order to use strategically limited spatial resources.

Elective surgery for liver injury and misinserted tube into the inferior vena cava associated with chest tube replacement: A case report.

Background: Several reports on organ injury and death due to incorrect chest tube insertion exist; however, reports on the chest tube penetrating the liver and reaching the inferior vena cava are limited. Case Presentation: A 79-year-old man presented with a clamped tube because of massive bleeding from the tube following right chest tube replacement in the hospital of origin. The tube entered the inferior vena cava from the hepatic parenchyma via the right hepatic vein and was removed 15 h later because his hemodynamics stabilized. A ruptured pseudoaneurysm necessitated further transcatheter arterial embolism on the second hospitalization day, and the patient was transferred back to the referring hospital on day 17. Conclusion: Liver injury caused by an inferior vena cava misinsertion-associated chest tube can be treated with elective surgery in anticipation of the tube's tamponade effect. However, due to the risk of rebleeding, imaging follow-up is necessary soon after surgery.

Upper-Arm SBP Decline Associated with Repeated Cuff-Oscillometric Inflation Significantly Correlated with the Arterial Stiffness Index.

We evaluated the clinical significance of the new non-invasive vascular indices to explore their potential utility using repeated cuff-oscillometric inflation. In 250 consecutive outpatients, we performed a cross-sectional, retrospective, single-center, observational study to investigate sequential differences in arterial stiffness using blood pressure, arterial velocity pulse index (AVI), and arterial pressure volume index (API) with repeated measurements. Males accounted for 62.7% of the patients, and the mean age was 68.1 ± 12.1 years. The mean systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the first reading in repeated measurements were 133.07 ± 21.20 mmHg and 73.94 ± 13.56 mmHg, respectively. The mean AVI and API were 23.83 ± 8.30 and 31.12 ± 7.86, respectively. In each measurement of these parameters, although DBP and AVI did not show significant changes throughout repeated measurements, SBP and API decreased significantly according to the measurement orders. Furthermore, changes in SBP and API were significantly correlated in several of the models. In this study, it was concluded that upper-arm SBP decline associated with repeated cuff-oscillometric inflation was significantly correlated with the arterial stiffness index. The findings of this study will allow clinicians to easily recognize the progression of atherosclerosis through regular, routine practice. In conclusion, this study suggests that changes in repeated SBP measurements may be predictive of arterial stiffness and atherosclerosis.

Development of a liquid chromatography-tandem mass spectrometry method for simultaneous quantification of hen's egg white allergens Gal d 1-4 in fresh and processed eggs.

Hen's egg white allergens, namely Gal d 1-4, cause food allergies worldwide and their intake must be strictly controlled by allergic individuals. However, an efficient method for quantifying these allergens is currently unavailable. We aimed to develop an LC-MS/MS method for simultaneous Gal d 1-4 quantification. Purified Gal d 1-4 proteins were trypsin-digested and the resulting peptides used in LC-MS/MS analysis. The limits of quantification were 9.77-39.1 ng/mL. The Gal d 1-4 recovery in fresh and processed eggs was 68.3-121.3%, and intra- and interassay coefficients of variation were 1.5-15.7% and 2.4-38.1%, respectively, indicating high sensitivity, accuracy, and reproducibility. In addition, the high specificity of this method was confirmed by testing 27 other foods. This newly developed method could provide reliable information to the industrial food and clinical fields, facilitating improved quality of life for individuals with egg allergies.

Rationale and Design of the Orencia Atherosclerosis and Rheumatoid Arthritis Study (ORACLE Arthritis Study): Implications of Biologics against Rheumatoid Arthritis and the Vascular Complications, Subclinical Atherosclerosis.

To explore the biological and immunological basis of human rheumatoid arthritis and human atherosclerosis, we planned and reported a detailed design and rationale for Orencia Atherosclerosis and Rheumatoid Arthritis Study (ORACLE Arthritis Study) using highly sensitive, high-throughput, human autoantibody measurement methods with cell-free protein synthesis technologies. Our previous study revealed that subjects with atherosclerosis had various autoantibodies in their sera, and the titers of anti-Th2 cytokine antibodies were correlated with the severity of atherosclerosis. Because rheumatoid arthritis is a representative autoimmune disease, we hypothesized that both rheumatoid arthritis and atherosclerosis are commonly developed by autoantibody-mediated autoimmune processes, leading to incessant inflammatory changes in both articular joint tissues and vessel walls. We planned a detailed examination involving carotid artery ultrasonography, measurements of adhesion molecules, such as ICAM-1 (intercellular adhesion molecule 1) and VCAM-1 (vascular cell adhesion molecule 1) for the evaluation of atherosclerosis progression, and high-throughput, high-sensitivity, autoantibody analyses using cell-free technologies, with detailed examinations of the disease activity of rheumatoid arthritis. Analyses of correlations and associations between biological markers and degrees of carotid atherosclerosis over time under consistent conditions may enable us to understand the biological and humoral immunity background of human atherosclerosis and autoimmune diseases.

Ability of suvorexant to prevent delirium in patients in the intensive care unit: a randomized controlled trial.

AIM: There are no effective, tolerable, and established medications for preventing delirium in critically ill patients admitted to the intensive care unit (ICU). We investigated whether suvorexant was effective in preventing ICU delirium. METHODS: This randomized controlled study evaluated 70 adult patients (age ≥20 years) admitted to the mixed medical ICU of the Tokyo Medical University Hospital (Tokyo, Japan) between May 2015 and February 2017. Patients were randomized using a sealed envelope method to receive either suvorexant (n = 34; 15 mg for elderly patients and 20 mg for younger adults) or conventional treatment (n = 36) for a 7-day period. The primary outcome was delirium incidence based on the definition in the 5th edition of the Diagnostic and Statistical Manual of Mental Disorders. RESULTS: No significant between-group differences were observed in the demographic or clinical characteristics. Kaplan-Meier estimates revealed that time to delirium onset was significantly longer in the suvorexant group than in the conventional group (P < 0.05). CONCLUSION: Suvorexant might be effective in preventing delirium in ICU patients.

Inhibition of inducible nitric oxide synthase ameliorates lung injury in rats after gut ischemia-reperfusion.

BACKGROUND: Gut hypoperfusion is considered to be a critical event for organ failure during severe surgical insults. The mechanism of remote organ injury after intestinal ischemia-reperfusion (I/R) may involve the excessive nitric oxide (NO) production; however, its role has been controversial. We sought to determine whether a selective inducible NO synthase inhibitor, aminoguanidine (AG), ameliorates pulmonary microvascular injury after superior mesenteric artery occlusion. METHODS: Anesthetized rats underwent superior mesenteric artery occlusion for 30 minutes and reperfusion for 6 hours (I/R) or sham operation (control). Another set of animals undergoing I/R received an AG at the end of the ischemia. Pulmonary vascular permeability was assessed by measuring tissue retention of Evans Blue dye that binds albumin. The plasma was harvested and NO2/NO3 (end products of NO) was measured. The bacterial cultures of the mesenteric lymph node of animals were performed to estimate the gut bacterial translocation after injury. RESULTS: The concentration of NO2/NO3 of plasma in the I/R group was higher than that of the control (p < 0.05). The lung-to-plasma Evans Blue dye ratio in the I/R group was also higher than that of the control (p < 0.01). Treatment with the AG prevented this lung injury induced by the gut I/R. The incidences of gut translocation were not significantly different between the I/R and AG groups. CONCLUSIONS: Increased lung vascular permeability elicited by gut I/R was significantly attenuated with inhibition of an inducible NO release by AG. Control of bacterial translocation was not needed to prevent lung injury in this model.

Monitoring of Antibiotic Residues in Aquatic Products in Urban and Rural Areas of Vietnam.

Antibiotic residues in aquatic products in Vietnam were investigated. A total of 511 fish and shrimp samples were collected from markets in Ho Chi Minh City (HCMC), Thai Binh (TB), and Nha Trang (NT) from July 2013 to October 2015. The samples were extracted with 2% formic acid in acetonitrile and washed with dispersive C18 sorbent. Thirty-two antibiotics were analyzed by LC-MS/MS. Of the 362 samples from HCMC, antibiotic residues were found in 53 samples. Enrofloxacin was commonly detected, at a rate of 10.8%. In contrast, samples from TB and NT were less contaminated: only 1 of 118 analyzed samples showed residues in TB and only 1 of 31 showed residues in NT. These differences were attributed to the local manufacturing/distribution systems. To understand the current status of antibiotic use and prevent adverse effects that may be caused by their overuse, continual monitoring is required.

Biodegradation of bisphenol A and disappearance of its estrogenic activity by the green alga Chlorella fusca var. vacuolata.

Bisphenol A (BPA) is known as an endocrine disruptor and often is found in landfill leachates. Removal of BPA by green alga, Chlorella fusca, was characterized, because we previously found that various phenols were well removed by this strain, including BPA. Chlorella fusca was able to remove almost all BPA in the concentration range from 10 to 80 microM for 168 h under continuous illumination at 18 W/m2. At the low light intensity of 2 W/m2, 82% of 40 microM BPA was removed, and only 27% was removed in the dark. Moreover, C. fusca could remove 90% of 40 microM BPA under the 8:16-h light:dark condition, which was almost as high as that under the continuous-light condition. The amount of BPA contained in the cells was less than the amount of BPA removed from the medium. Monohydroxybisphenol A was detected as an intermediate of BPA degradation. Moreover, estrogenic activity that originated from BPA in the culture medium also completely disappeared. Based on these results, BPA was finally degraded to compounds having nonestrogenic activity. Therefore, C. fusca can be considered a useful organism to remove BPA from landfill leachates.

Rapid and easy multiresidue method for the analysis of antibiotics in meats by ultrahigh-performance liquid chromatography-tandem mass spectrometry.

This study involved the development of a multiresidue method for the rapid analysis of 43 antibiotics in meats using ultrahigh-performance liquid chromatography-tandem mass spectrometry. This method was performed using dispersive-solid phase extraction, which is able to analyze 20 samples within 2 h. All compounds were determined simultaneously on a C18 separation column with gradient elution. Validation of the analytical method was performed by carrying out linearity, limit of quantification (LOQ), accuracy, precision, and recovery tests in different meat products. The validation criteria were set according to AOAC International and Japanese validation guidelines. The linearity of each compound was almost the coefficient of determination (r(2)) > 0.98. The LOQs of all tested antibiotics were <10 µg/kg. The results verify that this method is capable of quantitative analysis of 36, 33, and 37 compounds in beef, pork, and chicken, respectively. This method can be used for rapid and easy multiresidue screening of antibiotics for three meats (pork, beef, and chicken).

Hemostasis with emergently modified application of intra-aortic balloon occlusion in a patient with impending cardiac arrest following blunt proximal thigh amputation.

Case: A 30-year-old woman had her left thigh run over by a train. We tried to compress the left femoral area to control the arterial bleeding, but bleeding continued from the stump and injured soft tissue. The application of a tourniquet bandage also failed because of the limited remaining thigh. She developed impending cardiac arrest. As the left femoral arterial pulsation was still palpable, we inserted an intra-aortic balloon occlusion catheter percutaneously. The hemorrhage from the stump region decreased rapidly. She was transferred to an operating room to carry out surgical hemostasis, and it was confirmed with deflation of the balloon in the common iliac artery. Outcome: There was no complication of the skin or soft tissue at the surgical site caused by impaired circulation, and her consciousness fully recovered. Conclusion: We report the successful control of bleeding by the emergently modified application of intra-aortic balloon occlusion in the left common iliac artery.

Antibiotic residue monitoring results for pork, chicken, and beef samples in Vietnam in 2012-2013.

A monitoring plan of residual antibiotics in food of animal origin was conducted in Vietnam from 2012 to 2013. Meat samples were collected from slaughterhouses and retail stores in Ho Chi Minh City and Nha Trang. A total of 28 antibiotics were analyzed using a LC-MS/MS screening method. Sulfonamides, fluoroquinolones, and tilmicosin were detected in some of the samples. Sulfaclozine and fluoroquinolones were mainly detected in chicken samples, and sulfamethazine was mainly detected in pork samples. High levels of sulfonamide residues, ranging between 2500 and 2700 µg/kg sulfaclozine and between 1300 and 3600 µg/kg sulfamethazine, were present in two chicken and three pork samples, respectively. Tilmicosin was detected at ranges of 150-450 µg/kg in 10 chicken samples. Positive percentages were 17.3, 8.8, and 7.4% for chicken, pork, and beef, respectively, for an average of 11.9%. The results suggest an appropriate withdrawal period after drug administration had not been observed in some livestock.