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Excessive intake of caffeine, otherwise known to be a safe and mild central nervous system stimulant, causes nausea, vomiting, convulsions, tachycardia, and eventually fatal arrhythmias and death. Caffeine intoxication, a global problem, has been increasing in Japan since 2013. Thus, there is a need for rapid and accurate diagnosis of caffeine poisoning in forensic and clinical toxicology investigations. Herein, we demonstrate rapid and accurate caffeine quantitation by liquid chromatography tandem mass spectrometry using the standard addition method in a fatal case. Biological samples were diluted 500–100,000-fold and subjected to a simple pretreatment (adding caffeine standard and internal standard and passing through a lipid removal cartridge). The multiple reaction monitoring transitions were 195 → 138 for quantitation, 195 → 110 for the qualifier ion, and 204 → 144 for the internal standard (caffeine-d9). The standard plots were linear over 0–900 ng/mL (r2 = 0.9994–0.9999) for biological samples, and the reproducibility (%RSD) of the method was 1.53–6.97% (intraday) and 1.59–10.4% (interday). Fatal levels of caffeine (332 μg/mL) and toxic to fatal levels of olanzapine (625 ng/mL), along with other pharmaceuticals were detected in the external iliac venous blood. The cause of death was determined to be multi-drug poisoning, predominantly caused by caffeine. Our method is useful for not only forensic cases but also the rapid diagnosis of caffeine overdose in emergency clinical settings.

Background Bone union after spinal fusion surgery with instrumentation has been determined only with imaging studies. We evaluated the usefulness of the serum ionic fluoride (SIF) concentration as a biomarker of the bone union status. Methods We enrolled 25 patients who underwent spinal surgery in our institution, and we divided patients into three groups with and without instrumentation (G1, G2, and G3). We collected the fasting serum level preoperatively and on day 1 (D1), week 1 (D7), week 2 (D14), month 1 (D30), month 3 (D90), and month 6 (D180) postoperatively, and measured SIF concentrations using the flow injection method with an ion-selective electrode. Results Although preoperative SIF concentrations were similar among the 3 groups, postoperative SIF concentrations were different among the groups. SIF concentrations in groups with instrumentation (G2 and G3) increased between D14 and D90 postoperatively and decreased at D180 postoperatively. SIF concentrations in the group without instrumentation (G1) decreased between D30 and D180 postoperatively. Conclusions An SIF concentration that is higher postoperatively than preoperatively may indicate unstable bone union, whereas a lower SIF concentration postoperatively than preoperatively may indicate stable bone union. We concluded that the SIF concentration may be useful for diagnosing bone union.

Objective We conducted a retrospective study on the epidemiological and clinical features of patients with acute caffeine poisoning in Japan. Methods Letters requesting participation were sent to 264 emergency departments of hospitals, and questionnaires were mailed to those that agreed to participate. Patients Participants were patients transported to emergency departments of hospitals between April 2011 and March 2016 after consuming large or massive amounts of caffeinated supplements and/or energy drinks (caffeine dose ≥1.0 g). Results We surveyed 101 patients from 38 emergency departments. Since April 2013, the number of patients has markedly increased. Of these young patients (median age, 25 years), 53 were men, and 97 had consumed caffeine in tablet form. Estimated caffeine doses (n=93) ranged from 1.2 to 82.6 g (median, 7.2 g). Serum caffeine levels on admission (n=17) ranged from 2.0 to 530.0 μg/mL (median level, 106.0 μg/mL). Common abnormal vital signs and laboratory data on admission included tachypnea, tachycardia, depressed consciousness, hypercreatinekinasemia, hyperglycemia, hypokalemia, hypophosphatemia, and hyperlactatemia. Common signs and symptoms in the clinical course included nausea, vomiting, excitement/agitation, and sinus tachycardia. Seven patients (6.9%) who had consumed ≥6.0 g of caffeine, or whose serum caffeine levels on admission were ≥200 μg/mL, developed cardiac arrest. Ninety-seven patients (96.0%) recovered completely, but 3 patients (3.0%) died. Conclusion The present analysis of data from more than 100 emergency patients revealed clinical features of moderate to fatal caffeine poisoning. We recommend highlighting the toxicity risks associated with ingesting highly caffeinated tablets.

Analyses of drugs and poisons in tissue samples are essential in forensic toxicology and pharmacology. However, current procedures for tissue analysis are laborious and time-consuming. Therefore, we assessed the utility of a newly devised probe electrospray ionization (PESI) technique with tandem mass spectrometry (MS/MS) for easy, ultra-rapid drug detection in human tissue samples. Using this system, typical pretreatment procedures, such as solid-phase extraction, liquid-liquid extraction, deproteinization, or homogenization, can be avoided. Briefly, a tissue sample of 1-2 mm3 was supplemented with a solution of ethanol and 10 mmol/L ammonium formate, and measurements were obtained. We demonstrated the successful application of this method in a forensic case by detecting an opioid analgesic, MT-45, in all tissue samples (liver, kidney, lung, brain, and heart). We also detected oxidized metabolites of MT-45 in the liver. Since the analysis required only 0.5 minutes per sample, PESI-MS/MS is an ultra-rapid detection method. Furthermore, for a quantitative approach, the total analysis time for the combination of PESI-MS/MS with the quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction method (from instrument start-up to extraction and PESI-analysis) was within 8 minutes. MT-45 concentrations obtained by QuEChERS-PESI-MS/MS and liquid chromatography (LC) -MS/MS were similar for all tissue samples. PESI-MS/MS cannot be used to separate isobars/isomers (ie, compounds with the same m/z value), similar to other direct introduction techniques. Further studies are needed to validate the quantitation method. However, our results indicate that PESI-MS/MS is a potentially easy and rapid technique for the analysis of drugs and poisons in human tissue samples.

Case Is fomepizole necessary after massive ingestion of a mixture of methanol and ethanol? We report the case of a 37-year-old man who was transported to our Poison Center 12 h after ingesting 500 mL of fuel alcohol containing 70% methanol and 30% ethanol in a suicide attempt. On admission, he presented only with somnolence and mild metabolic acidosis. We hypothesized that most of the ethanol had been metabolized. Outcome As the estimated serum concentration of methanol was lethal (242.6 mg/dL), fomepizole was given i.v. and hemodialysis was carried out twice, resulting in complete recovery. Later, the serum concentrations of both methanol and ethanol on admission were found to be 224.1 and 0.51 mg/dL, respectively. Conclusion Therapeutic intervention was delayed by half a day after ingestion of a product containing methanol and ethanol in the present case. If the patient had arrived earlier, he may have only been treated with hemodialysis, but not fomepizole.

An easily obtainable thiol-selective labeling reagent [18F]FBSEM (N-[2-(4-[18F]fluoro-N-methylbenzenesulfonamido)ethyl]maleimide) was developed. The advantage of the design is that the precursor and [18F]FBSEM have the same backbone and backbone construction is not required; in contrast, known thiol-specific labeling reagents do require backbone construction, and this is thought to be the cause of their complicated synthesis. [18F]FBSEM was successfully obtained in higher yield (25%) and in a simpler way (two fluorination and deprotection steps in 65 min) than the widely used [18F]FBEM (N-[2-(4-[18F]fluorobenzamide)ethyl]maleimide). The labeling efficacy of [18F]FBSEM was confirmed by conjugation with glutathione. [18F]FBSEM is a promising labeling agent for proteins.

Glufosinate and glyphosate, which are non-selective herbicides that include an amino acid moiety in their structures, are frequently used worldwide to control unwanted vegetation. Unfortunately, these readily available herbicides are also used by people to commit suicide, and thus represent important chemicals of interest in the fields of clinical medicine and forensics. Because of the high water solubility of these herbicides, most analytical methods for their detection require a derivatization step, which results in longer analysis times. Therefore, derivatization-based methods do not currently contribute to judgements on treatment decisions in emergency medicine. In this study, we addressed this limiting factor by developing an ultra-rapid and simple analytical technique using a combination of probe electrospray ionization (PESI) and tandem mass spectrometry (MS/MS), which gives quantitative results within 0.3 min. Herbicide standards were added to human serum that was then subjected to analysis (N = 5 per concentration). The analysis was repeated daily over eight consecutive days. The limit of detection (LOD) was 0.59 μg/mL for glufosinate and 0.20 μg/mL for glyphosate. The limit of quantitation (LOQ), i.e., the lowest point on the calibration curves, was 1.56 μg/mL for both the herbicides. The matrix effects were observed at three different concentrations (between 95.7%–104% for glufosinate, and between 90.7%–95.7% for glyphosate). When applied to samples taken from actual poisoning cases (six samples for each herbicide), the present method gave almost the same quantitative values as those obtained by conventional high-performance liquid chromatography with fluorescence detection. Thus, we believe that PESI-MS/MS could emerge as a rapid diagnosis method in the clinical emergency field.

Poisoning incidents caused by drugs, accidental ingestion of poisons, attempted suicide, homicide, and exposure to toxic compounds occur frequently every year across the globe. This raises the need to rapidly identify toxic agents in poisoned patients in a clinical emergency setting. In addition, determining drug/poison concentration is undoubtedly necessary to arrive at a toxicological treatment plan. The purpose of this study was to develop an ultra-rapid drug screening method for the clinical treatment of poisoning. Probe electrospray ionization (PESI), one of the ambient ionization techniques, is able to detect compounds from various biological materials almost directly. We applied the PESI technique to the rapid detection of acetaminophen (APAP). Blood serum samples were diluted 100-fold with 10 mM ammonium formate/ethanol (1:1 v/v) solution including deuterium-labeled internal standards (IS; APAP-d4). Only 10 μL of the diluted sample was used for measurement. The tandem mass spectrometer (MS/MS) equipped with a PESI was used in selected reaction monitoring mode for the quantitation of APAP; the measurement time was only 18 s. Transitions were set at 152 > 110 for quantitation, 152 > 65 for qualifier, and 156 > 114 for IS (APAP-d4). All measurements were conducted in positive mode. The calibration curve (1/x2) was linear over the range of 1.56–200 μg/mL (r2 = 0.998), and the limit of detection and quantitation were 0.37 μg/mL and 1.56 μg/mL, respectively. The accuracy (bias) and precision (RSD%) of the method were within an acceptable range (−0.15–2.8% and 2.3–6.1%, respectively) and matrix effect at 3 concentrations (95.1–104%) indicated that PESI-MS/MS is only slightly affected by matrices. In real forensic cases, quantitative values of APAP determined by the PESI-MS/MS were almost identical to those determined by the liquid chromatography-MS/MS method. Since PESI-MS/MS is a simple, reliable, and rapid determination method for toxic agents with virtually no need for blood serum pre-treatment, it would be highly suitable for poisoning cases in clinical emergency settings. In the future, a method for simultaneous rapid determination of multiple toxic agents will be developed. Keywords: Acetaminophen, Blood serum, Probe electrospray ionization, Rapid drug screening, Tandem mass spectrometry

To the editorGlyphosate is used throughout the world as a non-selective herbicide. However, there are many reports of poisoning caused by glyphosate products, and ingestion of massive quantities of...

Background: The common garden plant autumn crocus contains colchicine and its derivatives. Colchicine poisoning causes abdominal cramps and diarrhea within hours. Several days later, multiorgan failure, pancytopenia, and cardiovascular collapse occur. 1,2 Severe colchicine poisoning is associated with high mortality. Case report: A 63-year-old woman who accidentally ingested an autumn crocus, which contained ~0.38 mg colchicine, had severe vomiting and was taken to an emergency center. She presented with symptoms of gastroenterocolitis within 1 hour of ingestion, and bone marrow hypoplasia with pancytopenia developed on the 3 rd day after ingestion. We continued administration of granulocyte colony-stimulating factor (300 mg) for 5 days until we confirmed that the patient's white blood cell count was increasing. Also, there was focal and segmental intestinal ischemia and some cakes of charcoal remained in the intestinal tract. Therefore, we presumed that nonocclusive mesenteric ischemia was caused by hypotension with severe dehydration, although pseudo-obstruction due to the activated charcoal may have been a contributing factor. We were able to promptly intervene to treat paralytic ileus and gastrointestinal edema before anticipated worsening of abdominal compartment syndrome, by conducting open peritoneal drainage. Despite severe poisoning, our patient survived with intensive care. Conclusion: Colchicine intoxication may lead to a sudden and extreme critical course. Therefore, as there is no means to predict prognosis from initial severity of symptoms at onset, we suggest that all patients suspected of colchicine intoxication should be managed in hospital with continuous vital sign monitoring and frequent laboratory testing for at least a few days after ingestion. Copyright © 2015, Taiwan Society of Emergency Medicine. Published by Elsevier Taiwan LLC. All rights reserved.

Aconitum and Colchicum are known to be highly toxic plants based on their respective toxins, namely aconite and colchicine. Aconite and colchicine poisoning generally occurs because of misidentification and contamination, and is also associated with homicide and suicide cases. The detection of aconitines and colchicine in body fluids and leftovers plays an important role in clinical and forensic toxicology investigation of suspected food poisoning by natural poisons. Therefore, we have developed and validated a rapid determination of aconitine, hypaconitine, jesaconitine, mesaconitine, and colchicine in serum, urine, and plants using liquid chromatography–tandem mass spectrometry. All targeted compounds, together with internal standards, were extracted from serum and urine using a C18 monolithic spin column. Recovery of all compounds from serum, spiked at concentrations of 0.1, 0.8, 6, and 18 ng mL−1, ranged from 96.8 to 101.4 %. Recovery of all compounds from urine, spiked at concentrations of 0.1, 8, 80 and 190 ng mL−1, ranged from 96.6 to 101.9 %. The intra- and inter-assay accuracy and precision (expressed as relative standard deviation, %RSD) evaluated for all compounds were within 96.7–103.2 % and 1.7–11.5 %, respectively. Detection limits and quantitation limits of aconitines and colchicine for serum and urine were 0.05 and 0.1 ng mL−1, respectively. This method had a chromatographic run time of 7.0 min. Calibration curves for serum and urine ranged from 0.1 to 20 ng mL−1 for serum and 0.1–200 ng mL−1 for urine, respectively. The validated method was successfully applied to clinical aconitine and colchicine poisoning cases.

Introduction Paraquat (PQ) is one of the most toxic herbicides to humans. However, it is still in use in many countries, including Japan, and many incidents, such as homicides, intentional ingestions, and occupational accidents, have been reported thus far. In PQ poisoning cases, it is possible to predict severity and prognosis using nomograms. Therefore, if the serum PQ level is determined immediately, a treatment plan can be rapidly established. However, most known analytical methods are time-consuming and therefore hardly ever contribute to patient treatment. Methods We developed a new method for PQ quantitation in serum by combining a probe electrospray ionization technique with mass spectrometry. This method requires virtually no serum pretreatment and can yield quantitation values in 18 s. Results We applied the proposed method to samples from real poisoning cases and compared the results with those obtained via liquid-chromatography–tandem mass spectrometry, revealing the absence of any significant differences at the 5% significance level (t(8) = 1.000, p > .05). The limits of detection and quantitation were 0.004 and 0.015 μg/L, respectively, and the calibration curve exhibited good linearity over the concentration range of 0.015–4.0 μg/mL (r2 = 0.998). Discussion As the proposed method is fast and easy to perform, it should be useful in emergency medical settings.

In acute glufosinate poisoning, sudden respiratory arrest and convulsion can occur after a latent period of 4-60 h. There is still no factor that accurately predicts the occurrence of these symptoms.To elucidate the predictors of severe effects following acute glufosinate poisoning.This study is a retrospective observational case series. The subjects were 16 patients who had acute glufosinate poisoning. They were divided into a group with respiratory arrest or convulsion during hospitalization (severe group) and a group without (non-severe group). The following characteristics (or predictors) were compared between the groups: age, sex, calculated amount of glufosinate (volume of ingested poison (glufosinate-containing herbicide) × glufosinate concentration of the product), time duration from poison ingestion to arrival at our hospital, use of gastric lavage, use of whole bowel irrigation, Glasgow Coma Scale, laboratory parameters, PaO₂/FiO₂ ratio (P/F ratio), shock index, and presence or absence of systemic inflammatory response syndrome (SIRS) on arrival.The P/F ratio was significantly lower in the severe group than in the non-severe group (median, 287.5 vs. 409.0; P = 0.049). The receiver operating characteristic (ROC) curve was plotted for the predictor of increasing severity based on the P/F ratio. The area under the curve was 0.714, and the optimal cutoff point for increasing severity was 374.0. The sensitivity was 75.0%, specificity of 71.4%, and accuracy of 75.0%. The shock index was significantly higher (median, 0.52 vs. 0.41; P = 0.031). Significantly more patients had SIRS in the severe group than in the non-severe group (P = 0.015). Logistic regression analysis was performed with a backward elimination procedure. SIRS was selected as the independent predictor of increasing severity (odds ratio, 29.810; 95% confidence interval, 1.011-878.952; P = 0.049).Severe effects following acute glufosinate poisoning were associated with two positive SIRS criteria. A low P/F ratio may be useful for predicting the occurrence of respiratory complications.

Aconite is a well-known toxic-plant containing Aconitum alkaloids such as aconitines, benzoylaconines, and aconins. We describe here the distribution of Aconitum alkaloids detected by liquid chromatography-tandem mass spectrometry (LC/MS/MS) in three autopsy cases of suicide by aconite poisoning. Case 1: a male in his fifties had eaten aconite leaves. The concentrations of jesaconitine in cardiac blood, urine, and kidney were 12.1 ng/ml, 993.0 ng/ml, and 114.2 ng/g, respectively. Case 2: a female in her fifties had eaten aconite root. The aconite root in the stomach included a high level of mesaconitine. The concentrations of mesaconitine in cardiac blood, liver, and kidney were 69.1 ng/ml, 960.9 ng/g, and 776.9 ng/g, respectively. Case 3: a male in his sixties had drunk liquor in which aconite root had been soaked. The concentrations of mesaconitine and aconitine in cardiac blood were 259.5 and 228.5 ng/ml, respectively. The Aconitum alkaloid levels were very high in the liver. The absorption of ethanol and Aconitum alkaloids might have been increased because of his having undergone total gastrectomy. In all three cases, the Aconitum alkaloid levels were high in the liver and kidney and low in the heart and cerebrum. The level in the cerebrum was lower than that in blood. Data on the distribution of the Aconitum alkaloids in the body in cases of aconite poisoning is useful to elucidate various actions of aconite alkaloids.

A patient committed suicide with hydrogen sulfide (H(2)S) by combining two commercial products. The patient was given hydroxocobalamin as an antidote in addition to treatment with cardiopulmonary resuscitation, but died approximately 42 min after his arrival at the hospital. The patient's cause of death was attributed to acute hydrogen sulfide poisoning. Serum concentrations of sulfide before and after administration of hydroxocobalamin were 0.22 and 0.11 μg/mL, respectively; serum concentrations of thiosulfate before and after hydroxocobalamin administration were 0.34 and 0.04 μmol/mL, respectively. Hydroxocobalamin is believed to form a complex with H(2)S in detoxification pathways of H(2)S. Although H(2)S is rapidly metabolized and excreted, the decreased sulfide concentration may be also associated with this complex formation. The decreased sulfide concentration suggests that hydroxocobalamin therapy may be effective for acute H(2)S poisoning. The decreased thiosulfate concentration seems to be associated with formation of a thiosulfate/hydroxocobalamin complex, because hydroxocobalamin can form a complex with thiosulfate. The thiosulfate concentration decreased to a greater extent than did sulfide, suggesting that hydroxocobalamin has a higher affinity for thiosulfate than for H(2)S. Therefore, prompt administration of hydroxocobalamin after H(2)S exposure may be effective for H(2)S poisoning.

The aconite tuber, which is processed from the roots of a flowering plant commonly known as monkshood, is used clinically for its analgesic, thermal, metabolic, cardiac, and diuretic effects [1]. The aconite tuber contains diterpenoid alkaloids, including aconitine, mesaconitine, hypaconitine, and jesaconitine, and mesaconitine plays a central role in the analgesic effects of this tuber [2]. Powdered aconite root is subjected to toxicity-reducing procedures that are collectively termed “processing.” It is believed that Aconitum alkaloids cannot be detected in the serum of patients who receive Chinese herbal medicines containing processed aconite root [3e6]. Previously, we developed a method to detect four types of Aconitum alkaloids in cases of aconitine poisoning, by using liquid chromatography-mass spectrometry (LC-MS) techniques, and we applied this method in a clinical setting [7e9]. Benzoylaconine, benzoylmesaconine, benzoylhypaconine, and 14-anisoylaconine are formed at the C-8 site during the toxicityreducing process following acetylic hydrolysis of Aconitum alkaloids. We previously detected four types of Aconitum alkaloids, aconitine, mesaconitine, hypaconitine, and jesaconitine, in healthy adult volunteers who were administered powdered processed aconite root containing Aconitum carmichaeli Debeaux and Aconitum japonicum Thunberg mixed at a fixed ratio [10]. Several pharmaceutical companies produce their own powdered processed aconite root products that may contain different varieties of aconite

Isolation of algicidal compounds from Ulva fasciata revealed that the algicidal substances were the polyunsaturated fatty acids (PUFAs) as hexadeca-4,7,10,13-tetraenoic acid (HDTA) C16:4 n-3, octadeca-6,9,12,15- tetraenoic acid (ODTA) C18:4 n-3, α-linolenic acid (ALA) C18:3 n-3 and linoleic acid (LA) C18:2 n-6. The fatty acid composition of four species of Ulvaceae (U. fasciata, U. pertusa, U. arasakii and U. conglobota) was analyzed by capillary gas chromatography to investigate the relationship with the algicidal activity. The results indicate that highly algicidal species, U. fasciata and U. pertusa, showed higher contents of C16:4 n-3, C18:3 n-3, and C18:4 n-3. Concentrations of these PUFAs released from the seaweed in the culture medium were also analyzed. These PUFAs were found to be significantly active against Chattonella antiqua, C. marina, Fibrocapsa japonica, Heterosigma akashiwo, Karenia mikimotoi, moderately effective against Heterocapsa circularisquama, Prorocentrum minimum, P. sigmoides, Scrippsiella trochoidea, whereas low effective against Alexandrium catenella and Cochlodinium polykrikoides. It is suggested that the PUFAs are useful mitigation agents to remove several harmful effects without causing detrimental effects on surrounding marine living organisms., Journal of Applied Phycology, 20(5), pp.713-720; 2009

Background: MicroRNA (miRNA) molecules have been detected in many body fluids and used as biomarkers. However, blood cell-derived miRNA molecules due to hemolysis have been shown to affect the amounts of plasma miRNAs. It is important to check the quality of plasma samples for clinical applications. In the present study, an objective method for quality checking plasma samples was investigated using permitted color tone level and the absorbance at 414 nm. Material and method: An ROC analysis of the macroscopic color tone and the absorbance in 1213 clinical samples was performed. The optimal cut-off absorbance value was validated using the amount of plasma miRNA. Results: AUC for detecting hemolyzed samples was very high (0.986) at a cut-off absorbance value of 1.664. A sensitivity and specificity were 99% and 92%, respectively. For validation study, 3 candidate miRNAs were selected by a miRNA microarray between fresh and hemolyzed plasma samples; the amounts of miR-16 and miR-19b increased in hemolyzed samples, whereas that of miR-223 remained unchanged. The amounts of plasma miR-16 and miR-19b were significantly increased in 5 clinical samples with higher absorbance values (p=0.0001 and p=0.0003, respectively), while the amounts of these miRNAs were not increased in samples with lower absorbance values. Conclusions: A simple method for quality checking plasma samples using color tone and absorbance is very useful and significant.

Alpha-linolenic acid and linoleic acid isolated from Ulva fasciata showed toxic effects on red tide phytoplankters in a concentration-dependent manner. Among six species tested, raphidophycean flagellate Heterosigma akashiwo was the most susceptible to these fatty acids, and 50% lethal concentrations (LC50) of alpha-linolenic acid and linoleic acid were estimated to be 0.58 and 1.91 microg/ml respectively, whereas dinoflagellate Gymnodinium impudicum and Heterocapsa circularisquama were highly resistant and no significant toxic effects were observed up to 1,000 microg/ml. Both fatty acids were less toxic to fish (devil stinger), zooplankters (brine shrimp and rotifer), and mammalian cell lines (U937, HeLa, Vero, and CHO cells) than H. akashiwo.

AIM: To evaluate the effectiveness of barium impaction therapy for patients with colonic diverticular bleeding. METHODS: We reviewed the clinical charts of patients in whom therapeutic barium enema was performed for the control of diverticular bleeding between August 2010 and March 2012 at Yokohama Rosai Hospital. Twenty patients were included in the review, consisting of 14 men and 6 women. The median age of the patients was 73.5 years. The duration of the follow-up period ranged from 1 to 19 mo (median: 9.8 mo). Among the 20 patients were 11 patients who required the procedure for re-bleeding during hospitalization, 6 patients who required it for re-bleeding that developed after the patient left the hospital, and 3 patients who required the procedure for the prevention of re-bleeding. Barium (concentration: 150 w%/v%) was administered per the rectum, and the leading edge of the contrast medium was followed up to the cecum by fluoroscopy. After confirmation that the ascending colon and cecum were filled with barium, the enema tube was withdrawn, and the patient’s position was changed every 20 min for 3 h. RESULTS: Twelve patients remained free of re-bleeding during the follow-up period (range: 1-19 mo) after the therapeutic barium enema, including 9 men and 3 women with a median age of 72.0 years. Re-bleeding occurred in 8 patients including 5 men and 3 women with a median age of 68.5 years: 4 developed early re-bleeding, defined as re-bleeding that occurs within one week after the procedure, and the remaining 4 developed late re-bleeding. The DFI (disease-free interval) decreased 0.4 for 12 mo. Only one patient developed a complication from therapeutic barium enema (colonic perforation). CONCLUSION: Therapeutic barium enema is effective for the control of diverticular hemorrhage in cases where the active bleeding site cannot be identified by colonoscopy.

SUMMARY Ethyl methane sulfonate-induced mutagenesis techniques were used to obtain cerulenin and erythromycin resistant mutant strains, which show a marked increase of 29 and 12%, respectively, in eicosapentaenoic acid content compared to the wild type (WT) strain. The increase in eicosapentaenoic acid content in both strains correlated well with a concomitant increase in 14:0, 16:0, 16:1, 18:0 and 20:4 n − 6 content of fatty acids. However, the linoleic acid content increased only in the cerulenin resistant strain, whereas the erythromycin resistant strain was completely devoid of this fatty acid, providing a contrasting feature for further study. Partial characterization of the mutant strains based on whole cell ultrastructure, growth patterns with or without selection pressure as well as at different temperatures revealed that the mutants were more thermotolerant, and maintained their morphological and genomic fidelity, although minor changes in band patterns were noted with random amplified polymorphic DNA techniques compared to the WT strain. Therefore, the developed new strains could have wider applications as planktonic feed for the mariculture industry. Further characterization of the mutants at the molecular level revealed that steady-state mRNA transcript levels of chloroplast-localized Δ12 desaturase and stearoyl-ACP desaturase genes showed a contrasting transcript profile. However, the level of endoplasmic reticulum-localized Δ12 desaturase mRNA transcript in either of the mutant strains was almost equal to that of the WT strain. Based on this observation, we postulate that the complete loss of 18:2 fatty acid content in the erythromycin resistant strain might be a result of mutation in the chloroplast-localized Δ12 desaturase gene and that the increased content of very long chain-polyunsaturated fatty acids might result from extrachloroplastic lipid desaturase activity, a phenomenon similar to that in the majority of angiosperms.

We developed a method for the determination of milnacipran in serum using a system combining liquid chromatography and mass spectrometry (LC/MS) in which electrospray ionization (ESI) was used. Milnacipran was extracted from serum by liquid-liquid extraction using an Extrelut® NT 3 column. The chromatographic separations of milnacipran and amoxapine (internal standard) were performed using a XTerra® RP 18 (150 × 2.1 mm I.D.) column with a gradient of 0.1% formic acid and 0.1% formic acid in acetonitrile as the mobile phase. The extraction recoveries at concentrations of 10 and 100 ng/mL were 85.6 and 83.7%, respectively. The calibration curve was linear in a concentration range from 5 to 300 ng/mL and the detection limit was 1 ng/mL. The intra-day and inter-a day assay precision expressed as coefficients of variation (C.V.) were 1.94-2.17% and 5.75-6.20%, respectively. This method proved to be useful in quantifying milnacipran in serum obtained from a patient who took this drug.

ポリプロピレン樹脂はその優れた特性が時代のニーズともよくマッチし,幅広い分野で用いられている。ポリプロピレンの高性能化には分子設計技術・成形加工技術を駆使した高次構造の制御が不可欠である。その研究の事例と今後のさらなる高性能化の可能性を紹介する。

Diarylethene derivatives having one or two chryso[b]thiophene ring(s) have been synthesized. In solution the derivatives underwent photochromism and the derivative with one chryso[b]thiophene ring exhibited photochromism even in the single crystalline phase. The color of the UV irradiated single crystal changed from blue to green by rotation the sample under polarized light.

We investigated a portable bioartificial renal tubule device (BRTD) consisting of renal tubule cells and hollow fibers, to improve the quality of life of patients. It is necessary for a BRTD system to be compact. A compact portable BRTB requires transfection of an appropriate water channel or electrical pump genes in tubular epithelial cells, which should be based on physiological similarities to human kidney function. LLC-PK(1) cells, into which rat kidney aquaporin 1 (AQP1) cDNA was stably transfected, were evaluated for water transport ability. The expression and localization of water AQP1 were examined by Western blotting, RT-PCR, and immunofluorescence. To measure transcellular water permeation, a simple method was applied, using phenol red as a cell-impermeant marker of concentration. In contrast to wild-type LLC-PK(1) cells, rat AQP1-transfected cells had high transcellular osmotic water permeability. The expression of rat AQP1 mRNA (ratio of AQP1 to beta-actin mRNA) and protein bands (a 28-kDa band and a broad, 35- to 45-kDa band) was confirmed to be stably maintained until a population doubling level of 24. In AQP1-transfected LLCPK(1) cells, the protein was localized mainly to the basolateral side, but also the apical side, of the plasma membrane. Wild-type LLC-PK(1) cells were not stained at the plasma membrane. It is possible that enough AQP1-transfected tubule epithelial cells were supplied for a bioartificial renal tubule device.

Nannochloropsis oculata was subjected to N-methyl-N-nitrosourea-induced mutagenesis under the selection pressure of quizalofop, a known inhibitor of acetyl-CoA carboxylase (ACCase) activity with the objective of generating genetically tractable mutants with altered fatty acid metabolism. Two mutants, QUIZ1 and QUIZ2, with stable resistance to quizalofop were isolated and partially characterized. The growth properties and morphology of the mutants appeared identical with the parent strain. However thermo-tolerance was observed in the mutants. Enhanced resistance to quizalofop suggested the presence of herbicide resistant isoforms of ACCase. In vitro assays for ACCase activity showed that ACCase in the wild strains was much more sensitive to quizalofop than the mutant strains. Gas chromatographic analysis of fatty acids revealed that the mutant strains were rich in polyunsaturated fatty acids (n− 3PUFAs), as well as total fatty acid contents; this was accompanied by a concomitant increase in triacylglycerol (TAG) followed by linoleic acid (18:2), arachidonic acid (20:4 n− 6) and EPA (20:5 n− 3). These results suggest that an increased substrate pool (malonyl-CoA) (due to increased specific activity of ACCase) in the mutant strains in vivo and in vitro may have led to the increased TAG accumulation. Random mutagenesis was shown to be a good tool to manipulate PUFAs and EPA in Nannochloropsis. The strains developed will be useful in understanding fatty acid metabolism using genetic and biochemical approaches and also for their direct use in mariculture.

We investigated the crystalline morphology of isotactic polypropylene obtained by melt crystallization under carbon dioxide (CO2) at various pressures. Spherulites consisting of regularly arranged fibrils without subsidiary lamellae were obtained by crystallization under CO2 below 2 MPa, whereas large spherulites consisting of irregularly arranged fibrils with subsidiary lamellae were obtained under ambient pressure. Distorted domain crystals with uniform optical anisotropy consisting of α-form were found to be obtained under CO2 above 2 MPa, and needle crystals consisting of γ-form were obtained above 12 MPa. Transmission electron micrographs showed that straight and thick lamellae are regularly arranged in both the distorted domain crystals and the needle crystals. The uniformly thick lamellae were confirmed by differential scanning calorimetry thermograms; that is, the melting temperature is higher and the melting peak is sharper than those obtained under ambient pressure. Such characteristic crystalline morphologies obtained under CO2 may be attributed to local ordering in the melt state. © 2004 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 42: 2738–2746, 2004

To understand the cause of discoloration of the sea laver "nori," which is found in the Ariake Sea, the concentrations of pigments and elements in the normal and discolored laver samples were determined. In the discolored samples, a decrease in all of the pigments, chlorophyll a and carotenoids, and proteinous pigments, phycobiliproteins, was clearly observed. This was accompanied by a decrease in the content of Fe, Zn, Mn, Cu, and P. Good correlations between these elements and chlorophyll a, as well as between these elements and phycobiliproteins, were confirmed, indicating that, in addition to the deficiency of nitrogen and phosphorus, the deficiency of trace elements (Fe, Zn, Mn, and Cu), which are specifically required for photosynthesis, could be a reason for the discoloration of nori. The cause of elemental deficiency is also discussed.

Pythium porphyrae (Oomycota) is a microbial pathogen which causes red rot disease in the commercially cultivated red seaweed Porphyra. This disease is initiated by the motile zoospores of the fungus, which it has been suggested to recognize and process host specific signals by membrane bound receptors. Monoclonal antibodies (MAbs) were developed against the surface components of zoospores and cysts of this fungus in order to try and identify the putative receptor molecules involved in the zoospore encystment process. Screening of MAbs by immunofluorescence assays has revealed three different patterns of surface epitope binding, while labeling of zoospore and cysts components by FITC-conjugated lectins has identified different carbohydrate moieties. Of the MAbs and lectins tested, MAb 1A3 and wheat germ agglutinin have induced zoospore encystment under in vitro conditions. MAb 1A3 identified a 109 KDa band of a glycoprotein in western blot analysis which could be a putative receptor responsible for the induction of zoospore encystment.

The function of current hemodialysis as an artificial kidney is insufficient because of the lack of reabsorptive function. In this study, we intend to develop a bioartificial renal tubule cell device (RTD) using tubular epithelial cells and artificial membranes and to evaluate the reabsorptive function of the confluent layers. Madin-Darby canine kidney (MDCK) cells were cultured on a nucleopore polycarbonate membrane for up to 4 weeks after confluence to examine the influence of the culture period on their ability to transport Na+ actively using Na+/K+ATPase (NKA). The results were (1) active Na+ transport of the cells averaged 24.8 mM/m(2) x 24 h during the initial 2 weeks after confluence and then decreased to about 4.2 mM/m(2) x 24 h during the next 2 weeks; (2) NKA localized on the basal-lateral sides of the cells during the initial 2 weeks, whereas it also localized on the apical side of the cells during the next 2 weeks; (3) long-term culture resulted in an increased number of upheaving cell mass, increased fatty droplets in the cells, and necrosis; and (4) scanning electron microscopy showed fewer microvilli 3-four weeks after confluence. It is concluded that the culture period is critical for developing RTD using cultured renal tubular epithelial cells.

Pythium porphyrae Takahashi et Sasaki, a facultative parasite of Porphyra spp., is the common microbial agent responsible for red rot disease of this red alga in Japan. Host infection by this species and other plant parasitic members of the Pythiaceae is initiated by motile biflagellate zoospores. Factors regulating host specificity and the initial steps involved in the infection process, consisting of attachment, encystment and appressorium formation, are not known. Zoospore encystment and appressorium formation of P. porphyrae were monitored by staining of the fungal cell walls using calcofluor. The zoospores infected only Porphyra spp. and Bangia atropurpurea (Roth) C. Agardh thalli, although they attached to, and encysted on, many other members of the Rhodophyceae (Stylonema alsidii[Zanardini] Drew, Gelidium elegans Kutzing, Pterocladiella capillacea[Gmelin] Santelices et Hommersand, Carpopeltis affinis[Harvey] Okamura, Gloiosiphonia capillaris[Hudson] Carmichael in Berkeley, Grateloupia turuturu Yamada, Callophyllis adhaerens Yamada, Gracilaria spp., Lomentaria hakodatensis Yendo, Rhodymenia intricata[Okamura] Okamura, Griffithsia subcylindrica Okamura, Wrangelia tanegana Harvey, and Polysiphonia morrowii Harvey). No attachment or encystment was observed on the red alga Kappaphycus striatum (Schmitz) Doty ex Silva in Silva et al., the brown algae Undaria pinnatifida (Harvey) Suringar, Scytosiphon sp., and Sargassum thunbergii (Mertens ex Roth) Kuntze as well as members of the Ulvaceae (green algae). Sequential extraction of carbohydrates from Porphyra yezoensis Ueda thalli and the addition of diverse monosaccharides, polysaccharides, and amino acids to zoospore suspensions indicated that encystment and appressorium formation were induced only by sulfated galactans (porphyran, commercial agar, agarose, and carrageenans). Zoospore attachment and encystment on thalli of P. yezoensis was abolished by periodate oxidation of the thallus surface and was reduced by 80–90% after enzymatic removal of sulfated galactan (porphyran). It appears that the interaction of zoospore surface receptors with sulfated galactan (porphyran) determinants on the thallus surface induced specific attachment and encystment on Porphyra spp. thalli. Zoospores encysted, germinated, and formed appressoria on sulfated galactan films and in suspensions of this carbohydrate. Attachment and encystment were induced on commercial agar and agarose films, but appressoria were not induced on agarose films. Supplementation of agarose media with both cold and hot water fractions and with porphyran from P. yezoensis–induced appressoria implicated sulfated galactans (porphyran) in appressorium formation.

We intend to develop a bioartificial kidney using tubular epithelial cells and artificial membranes, and to evaluate the reabsorptive function of the confluent layers. Madin-Darby canine kidney (MDCK) cells were cultured on a nucleopore polycarbonate membrane for up to 4 weeks after confluence to examine the influence of culture period on their properties, such as the localization of Na+/K+-ATPase and active Na+ transport. The results were as follows. Ouabain-sensitive Na+ active transport declined at 3 to 4 weeks after confluence in each matrix. The localization of Na+/K+-ATPase indicated depolarization in the cell membrane 3 to 4 weeks after confluence. Prolongation of the culture period increased the formation of an upheaving cell mass after the formation of the confluent monolayer. Scanning electron microscopy revealed fewer microvilli and more flat cells after 3 to 4 weeks of confluency. We conclude that the decline of Na+ active transport in the MDCK cells was due to both the formation of multilayers and a decline of cell function throughout the long period of culture following the formation of the confluent monolayers. Further study for selection of membrane material, the extracellular matrix, and species of cells should be continued.

Advanced glycation and lipidoxidation of the peritoneal membrane: Respective roles of serum and peritoneal fluid reactive carbonyl compounds.BackgroundAdvanced glycation of proteins has been incriminated in the progressive alteration of the peritoneal membrane during chronic peritoneal dialysis (PD). Advanced glycation end products (AGEs) result from a modification of proteins by reactive carbonyl compounds (RCOs). RCOs resulting from glucose breakdown are present in commercial PD fluid. They also accumulate in uremic plasma. The present study was undertaken to evaluate the respective contribution of these two sources of RCOs in the genesis of peritoneal AGEs.MethodsThree major RCOs formed during heat sterilization of PD fluid, that is, glyoxal, methylglyoxal, and 3-deoxyglucosone, and total RCOs were measured in commercial PD fluid and in PD effluent. The generation of pentosidine, used as a surrogate marker for AGEs, during one-week incubations of PD fluid and effluent samples fortified with bovine serum albumin (BSA) was measured by high-performance liquid chromatography. Peritoneal samples were stained with antibodies specific for two AGEs derived from carbohydrate-dependent RCOs, Nε-(carboxymethyl)lysine (CML) and pentosidine, or for two advanced lipoxidation end products (ALEs) derived from lipid-dependent RCOs, malondialdehyde (MDA)-lysine and 4-hydroxynonenal (HNE)-protein adduct.ResultsGlyoxal, methylglyoxal, and 3-deoxyglucosone were identified in commercial PD fluid. Their levels in PD effluents decreased with dwell time probably by diffusion into blood circulation. In contrast, the levels of total RCOs were initially low in commercial PD fluid, increased in PD effluent with dwell time probably by diffusion from circulation into the peritoneal cavity, and after 12 hours, reached values observed in uremic serum. The relevance of the rise in total RCOs for AGE formation is demonstrated by a parallel increase in the generation of pentosidine during incubations of PD effluents. In contrast with RCOs present in glucose-rich PD fluid, RCOs diffusing from uremic circulation originate from both carbohydrates and lipids. Their role in the modification of peritoneal proteins is demonstrated by the immunohistochemical study of peritoneal tissue. Two AGEs and two ALEs increase in parallel in the mesothelial layers and in vascular wall of small arteries in the peritoneum.ConclusionsProtein modification of the peritoneum is determined not only by RCOs originating in PD fluid, but also by RCOs originating from the uremic circulation. The present data might be relevant to current attempts to improve PD fluid toxicity by lowering its glucose content.

Porphyra yezoensis Ueda conchospore germlings (1–4-cell stages) were treated with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) for inducing mutations. Three kinds of color-mutated gametophytic blades, which were composed of the mutated cells wholly, sectorially or spottedly, were obtained; and most of them were sectorially variegated blades. The highest frequency of these mutated blades was 1.3%. Four different pigmentation mutant strains were obtained by regenerating single cells and protoplasts that were enzymatically isolated from the mutated sectors of the sectorially variegated blades. The mutants were relatively stable in color in both gametophytic blade and conchocelis phases. In the two phases, each mutant strain showed characteristic differences in the in vivo absorption spectra, and had different pigment contents of major photosynthetic pigments (chlorophyll a, phycoerythrin and phycocyanin) as compared with the wild-type and with each other. The gametophytic blades from the four mutant lines showed significant differences in growth and photosynthetic rates, when they were cultured in the same conditions. By crossing the mutant with the wild-type, it was found that the color phenotypes of two mutants reported above, were resulted from two mutations in different genes, respectively.

It has been proposed that hematopoietic and endothelial cells are derived from a common cell, the hemangioblast. In this study, we demonstrate that a subset of CD34+ cells have the capacity to differentiate into endothelial cells in vitro in the presence of basic fibroblast growth factor, insulin-like growth factor-1, and vascular endothelial growth factor. These differentiated endothelial cells are CD34+, stain for von Willebrand factor (vWF), and incorporate acetylated low-density lipoprotein (LDL). This suggests the possible existence of a bone marrow-derived precursor endothelial cell. To demonstrate this phenomenon in vivo, we used a canine bone marrow transplantation model, in which the marrow cells from the donor and recipient are genetically distinct. Between 6 to 8 months after transplantation, a Dacron graft, made impervious to prevent capillary ingrowth from the surrounding perigraft tissue, was implanted in the descending thoracic aorta. After 12 weeks, the graft was retrieved, and cells with endothelial morphology were identified by silver nitrate staining. Using the di(CA)n and tetranucleotide (GAAA)n repeat polymorphisms to distinguish between the donor and recipient DNA, we observed that only donor alleles were detected in DNA from positively stained cells on the impervious Dacron graft. These results strongly suggest that a subset of CD34+ cells localized in the bone marrow can be mobilized to the peripheral circulation and can colonize endothelial flow surfaces of vascular prostheses.

Heparin is used as an anticoagulant drug. The anticoagulation process is mainly caused by the interaction of heparin with antithrombin followed by inhibition of anticoagulant factor IIa and factor Xa. The anti-factor IIa and anti-factor Xa activities of heparin are critical for its anticoagulant effect; however, physicochemical methods that can reflect these activities have not been established. Thus, the measurements of anti-IIa and anti-Xa activities by biological assay are critical for the quality control of heparin products. Currently in the Japanese Pharmacopoeia (JP), the activities of heparin sodium and heparin calcium are measured by an anti-Xa activity assay (anti-Xa assay), but anti-IIa activity is not measured. Here, we established an anti-IIa activity assay (anti-IIa assay) and an anti-Xa assay having good accuracy and precision. When samples having a relative activity of 0.8, 1.0 and 1.2 were measured by the established anti-IIa and anti-Xa assays in nine laboratories, good accuracy (100.0-102.8% and 101.6-102.8%, respectively), good intermediate precision (1.9-2.1% and 2.4-4.2%, respectively) and good reproducibility (4.0-4.8% and 3.6-6.4%, respectively) were obtained. The established anti-IIa and anti-Xa assays have similar protocols, and could be performed by a single person without a special machine. The established assays would be useful for quality control of heparin.

To evaluate the relationship between the metal and chlorophyll contents in dried sea lavers (nori), analyses of metals by ICP-AES and chlorophyll content by a Minolta chlorophyll meter, as expressed by the SPAD value, were performed. Seventy dried sea laver samples including color fading lavers were products from Ariake Sea. The correlations between the contents of Fe, Zn, Mn and the SPAD values was confirmed with the coefficient (R) being 0.774, 0.707 and 0.564, respectively. It was suggested that the SPAD value could be applicable to evaluate the quality of dried sea lavers.

Phase transformation of amorphous-silicon during millisecond annealing using micro-thermal-plasma-jet irradiation was directly observed using a high-speed camera with microsecond time resolution. An oval-shaped molten-silicon region adjacent to the solid phase crystallization region was clearly observed, followed by lateral large grain growth perpendicular to a liquid-solid interface. Furthermore, leading wave crystallization (LWC), which showed intermittent explosive crystallization, was discovered in front of the moving molten region. The growth mechanism of LWC has been investigated on the basis of numerical simulation implementing explosive movement of a thin liquid layer driven by released latent heat diffusion in a lateral direction.

A gram-scale miniature reactor designed for reactive blendind was introduced. A brief review was given on the light scattering method for morphology characterization and the ellipsometric analysis on polymer-polymer interface. These methods were applied for the reactive blending of poly(ethylene terephthalate) with ethylene-propylene rubber (70/30 PET/EPR). Compared with non-reactive system, the reactive one yielded finer dispersion of rubber particles. It showed better stability of particle dispersion during static annealing. The reactive system with thick interface showed higher impact strength than the non-reactive one having same particle size but thinner interface, suggesting that the thick interface may be a prerequisite to the rubber toughening.

Teeth are recently drawing attention for their potential as biological modeling investigation samples due to their ability to be collected and their slow substance metabolism. There is no active metabolism of elements after the completion of dentin. Dentin is surrounded by enamel and cementum, and is not affected by the oral environment. Therefore, the amount of trace elements in dentin may change with age, and this is considered to be a reliable biological load index. The objectives in this study are to demonstrate concentrations of elements in the dentin of healthy Japanese subjects by sex and age, and to reveal the relationship between element levels and age. 121 healthy teeth samples were extracted due to periodontal disease or orthodontic treatment. Each tooth was sliced from the crown to the root apex into 0.5–1 mm thickness, then enamel, cementum, and the pulp were removed; the dentins were used as samples. The concentration of 10 trace elements (B, Mn, Co, Cu, Zn, Rb, Sr, Mo, Cd, and Pb) in the dentin was measured using inductively coupled plasma mass spectrometer (ICP-MS). The differences Co and Pb in the dentin between men and women were significant (p

The physiological role of aryl hydrocarbon receptor (AhR) is not yet fully understood, and investigation is hampered by the limited solubility of reported AhR ligands in aqueous media. To achieve improved solubility, we focused on our previous finding that planarity-disruption of molecules leads to less efficient crystal packing and greater aqueous solubility. Here, we describe chemical modification of an AhR agonist, beta-naphthoflavone, focusing on planarity-disruption. As expected, introduction of substituents at the ortho-positions of the phenyl group resulted in greater solubility. Among the compounds prepared, the fluoro analog showed more potent AhR agonistic activity and greater solubility than did beta-naphthoflavone. Our results indicate that this strategy to improve aqueous solubility, that is, introduction of substituent(s) that disrupt planarity, may be generally applicable to bicyclic molecules.

Fusion of isolated protoplasts (using bacterial crude enzyme following 2% protease treat-ment) from the thalli of Porphyra yezoensis and P. pseudolinearis by the polyethylene glycol (PEG) and the electric stimulation methods is described. In the PEG method, both PEG solu-tion and PEG powder were tested in combination with different dilution media and compared for their fusion rates. The fusion rate of PEG solution and the powder methods did not differ (about 10%). However the electrofusion method yielded higher fusion rates than the PEG method. Optimum fusions were obtained in the protoplasts aligned in an AC field of 40V at 1MHz for 20s and subsequent fusion by a DC pulse of 300V for 40μs duration. Supplementation of Ca2+ and Mg2+ to the fusion buffer enhanced the fusion rates up to 32.6% (24.3% binary and trinary fusion products) under the same electrical conditions. Protoplasts isolated by bacterial enzyme had higher fusion frequencies than those isolated by using enzymes of top shell and abalone. All the three enzymes showed activity for porphyranase, β-1, 3-xylanase and β-1, 4-mannanase. However enzymes of top shell and abalone had lower activity for porphyranase and β-1, 3-xylanase than the bacterial enzyme.

Processed Bushi powder for ethical dispensing, called TJ-3022, is a herbal drug of processed Aconiti tuber (Aconitum carmichaeli Debeaux) and TJ-3023 is newly developed to contain a higher proportion of diester alkaloid of aconitine (Aconitum carmichaeli Debeaux and Aconitum japonicum Thunberg). Safety of TJ-3022 and TJ-3023 was evaluated by measuring the level of aconitum alkaloids in healthy adult volunteers. Forty subjects were assigned to one of two groups (each 20 subjects): TJ-3022 group or TJ-3023 group. The subjects received the powdered processed Aconiti tuber 3 g/day and the blood concentrations of aconitum alkaloids were measured at 90 min and 72 h after taking the study drug. The serum concentrations of aconitum alkaloids after 90 min and 72 h in the TJ-3023 group were higher than those in the TJ-3022 group. As for the chronological changes in the serum concentration, a significant decrease was observed in the TJ-3022 group, while no significant decrease was seen in the TJ-3023 group, which suggests that an analgesic effect in TJ-3023 was stronger than in TJ-3022. Aconitum alkaloids, which always have been believed to have the blood concentration below the measurement limit in human, were detected for the first time, although the detected amounts were minute. The results suggest that TJ-3023 shows sufficient analgesic effect with smaller dose than TJ-3022.

We have developed a simulation method to predict the ground deformation due to the dissociation of methane hydrate. In the dissociation process, the phase change from solid to fluids leads to the change in partial stresses in the porous media, which will cause the ground deformation. The simulations are based on the chemo-thermo-mechanical coupled finite element analysis, in which the phase change, the flow of pore fluids, the mechanical behavior of solid skeleton, and heat transfer are simultaneously solved. We treat the ground as unsaturated soils, and apply an elastoviscoplastic constitutive model to the soil skeleton. Using the proposed method, we have numerically analyzed the dissociation process for heating methods. Ground deformation has been predicted which is caused by water and gas generation during the dissociation.

Bioassay-guided fractionation of an ethanol extract of the brown alga, Dictyota dichotoma, led to the isolation of a novel chlorine-containing perhydroazulene diterpene together with two known diterpenes, dictyolactone and sanadaol. The structure of the novel compound, named dictyol J, was elucidated on the basis of spectroscopic information. Dictyolactone showed the highest algicidal activity against red-tide phytoplanktons among the three isolated compounds.