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Background: Various surgical techniques have been proposed to treat small aortic annulus. We have used small-diameter mechanical valves for small aortic annulus. The present study examined aortic valve replacement with small diameter mechanical valves to determine their effectiveness., Methods: The study included 67 patients who underwent elective, isolated aortic valve replacement with ATS-Advanced Performance (AP) valves (ATS Medical, Inc, Minneapolis, MN) within our department. The patients were divided into two groups; patients receiving the 16-mm AP valve (16-mm group) and those receiving valves 18-mm AP or larger (>16-mm group)., Results: The mean age of the 16-mm group was significantly higher than that of the >16-mm group. Body surface area was significantly smaller in the 16-mm group than the >16-mm group. We found no significant differences in preoperative conditions between the groups. The effective orifice area index of the mechanical valve was lower in the 16-mm group (0.87+/-0.06 vs 1.12+/-0.13 cm2/m2, p<0.0001). Six patients (40%) in the 16-mm group fell into the category of prosthesis-patient mismatch cases. Although the postoperative pressure gradient was higher in the 16-mm group (33.7+/-12.5 vs 23.16+/-8.78, p<0.01), left ventricular mass index and left ventricular ejection fraction at early and at late follow-up improved from the preoperative period in both groups. We found no significant differences in incidence of postoperative complications, including death., Conclusions: We believe aortic valve replacement with small-diameter mechanical valves (the ATS-AP valves) in patients with a small annulus provides satisfactory remote prognosis. The use of mechanical valves in elderly patients is considered acceptable with strict clinical monitoring to prevent thromboembolism and anticoagulation therapy-induced complications., (Copyright (c) 2010 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.)

Background: Daphnia magna is a useful aquatic organism for testing ecological toxicities of environmental pollutants. However, there were only a few studies on agricultural chemicals using these organisms., Methods: We investigated acute and subchronic toxicities of 30 agricultural chemicals commonly used in Japan in D. magna. Acute toxicity of the agricultural chemicals was determined using the concentrations yielding 50% immobility of D. magna after 24 hr and 48 hr exposure as end points. D. magna was cultivated with the chemical and algae until the first brood production. Lethal toxicity and the number of survival broods were determined within 13 days., Results: All insecticides among the agricultural chemicals exhibited the strongest acute toxicity (LC50 from 0.00053 to 0.037 mg/L). More than 50% of the herbicides and fungicides did not exhibit acute toxicity at 10 mg/L. Chlornitrofen, pencycuron, and fenitrothion showed significantly lower LC50 values at 8 days than at 24 hr and 48 hr. Isoprothiolane, flutolanil, and thiophanatemethyl significantly delayed the first brood at concentrations less than half of those for LC50 (8 days). Thiobencarb, iprodione, flutolanil, mepronil, and thiophanatemethyl significantly reduced the size of the first brood at concentrations less than half of those for LC50 (8 days)., Conclusions: In this study, chlornitrofen, pencycuron, and fenitrothion were suggested to have slow-acting toxicity. Also, thiobencarb, iprodione, flutolanil, mepronil, and thiophanatemethyl were suggested to have parthenogenetic toxicity.

Dimethylarsinous acid (DMAIII) induced tetraploidy and multinucleation in V79 cells. While the highest yield of tetraploids and multinucleated cells was at 1.25 microM of DMAIII, the mitotic index was highest at DMAIII 2.5 microM due to mitotic arrest. Mitosis was not observed at 5 microM. We observed V79 cells treated with DMAIII and visualized with rhodamine-phalloidin. Abnormal actin location was observed in the dividing cells which were treated with DMAIII and visualized with rhodamine-phalloidin. These findings suggested strongly that DMAIII inhibits not only formation of the normal mitotic spindle but cytokinesis and induces the formation of multinuclear cells.

A specific mercuric ion binding protein (MerP) originating from transposon TnMERI1 of Bacillus megaterium strain MB1 isolated from Minamata Bay displayed good adsorption capability for a variety of heavy metals. In this study, the Gram-positive MerP protein was expressed in transgenic Arabidopsis to create a model system for phytoremediation of heavy metals. Under control of an actin promoter, the transgenic Arabidpsis showed higher tolerance and accumulation capacity for mercury, cadium and lead when compared with the control plant. Results from confocal microscopy analysis also indicate that MerP was localized at the cell membrane and vesicles of plant cells. The developed transgenic plants possessing excellent metal-accumulative ability could have potential applications in decontamination of heavy metals.

The splicing of a bacterial group II subclass B intron B.me.I1 from Bacillus megaterium chromosomes was investigated. RT-PCR and nucleic acid hybridization methods were used to understand the role of the intron-encoded protein (IEP) in the splicing of B.me.I1. An in vivo assay showed that the splicing occurred in the absence of IEP. An in vitro assay showed that B.me.I1 was spliced under conditions similar to those of the intracellular environment with no help from other biological molecules. Because all group II introns previously reported needed IEPs for their splicing in vivo, our results suggest that B.me.I1 is an "actual" self-splicing group II intron. This is also the first report to recognize the existence of group II introns that independently splice mRNA in vivo. The self-splicing of a bacterial intron may support that eukaryotic spliceosomal introns originated in bacterial genomes.

Comparative analyses of bacterial community successions in the composting materials were done for a conventional windrow post-treatment (WPOT) process with the hyperthermophilic pre-treatment (HTPRT) and simple windrow composting (SWC; without the HTPRT). Multidimensional scaling profiles based on data of terminal restriction fragment length polymorphisms of the bacterial population in the samples of every 7 days composting material and analyses of the 16S rRNA gene-based clone library of the 7 and 21 days composting materials suggested that bacterial communities of the composting materials differed much between these two processes until the 35 days of composting, whereas that they were closely related to each other at the final composting stage (42 days of composting). Detailed phylogenetic analysis clarified that all WPOT clone libraries contained many clones of the lineages of aerobic bacteria (for example, bacilli). However, the most abundant clones retrieved from all SWC materials were affiliated with a clone cluster closely related to identified and classified members of the phylum Firmicutes that have strictly anaerobic metabolism pathways. From these results, we conclude that the HTPRT process contributed to easily establish an aerobic ecosystem from the early stage to the final stage of WPOT composting with plowing the materials only once a week.

The mercury resistance module of Bacillus transposon TnMERI1 is regulated by three operator/promoter regions (O/P merB3, O/P merR1, and O/P merR2) and two regulatory proteins (MerR1 and MerR2) encoded by the module itself. To clarify the roles of MerR1 and MerR2 in the regulatory mechanism, both proteins were overexpressed and purified. MerR1 bound the regulatory regions O/P merB3 and O/P merR1, with a preference for O/P merB3 as measured on in vitro gel shift assays. However, MerR2 bound O/P merR2, as revealed by gel shift and restriction endonuclease protection assays. The transcriptional start sites of O/P merB3 and O/P merR2 were determined by rapid amplification of 5'-cDNA ends (5'-RACE) in the TnMERI1 original host, Bacillus megaterium strain MB1. Real-time reverse transcription polymerase chain reaction (RT-PCR) assays showed that O/P merB3 and O/P merR1 were induced in the presence of Hg2+ but not O/P merR2. It was concluded that MerR1 regulates O/P merB3 and O/P merR1, while MerR2 regulates O/P merR2.

Transcription and mobilization facilities of a group II intron, B.me.I1, which was classified within a group IIB subclass and nested in a broad-spectrum mercury resistance transposon TnMERI1 found from the chromosome of Bacillus megaterium strain MB1 were investigated. Though B.me.I1 does not intervene in any recognizable exon gene, the splicing ability of B.me.I1 in Escherichia coli was confirmed by applying RT-PCR with RNA template transcribed by using a T7 RNA polymerase-promoter expression system. The homing activity of B.me.I1 was confirmed by using an intron-less allele fragment as a specific integration site which was cloned from Tn5084, a TnMERI1-family mercury resistance transposon. The transcription and splicing of B.me.I1 in the original host, the strain MB1, were also observed as its natural property. Primer-walking and 5' RACE analyses showed that the transcription start site and the putative promoter region of B.me.I1 were located on the antisense strand of tnpR gene of TnMERI1. Therefore, it is cleared that TnMERI1 provides a specific integration site and facilities for transcription of B.me.I1. From these results, it is considered that these genetic facilities given by the transposon TnMERI1 confer splicing and homing capability on B.me.I1, even though the group II intron is not associated with any exon gene.

Background: Arsenic in drinking water remains a major public problem in Bangladesh, although arsenic mitigation programs began there a decade ago. The purpose of this study was to examine the effectiveness of this program by determining the relationship between current arsenic levels in well water and the high level of urinary arsenic excretion., Methods: A community-based cross-sectional study was conducted in the Pabna district of Bangladesh between May and July 2005. We included 174 married couples and collected their drinking water from 138 wells. The allowable limit for arsenic in drinking water is 50 microg/L in Bangladesh, while the normal level of urinary arsenic is < or =40 microg x 1.5 L(-1) x day(-1) by Dhaka Community Hospital., Results: Of 348 subjects, 304 exceeded the urinary arsenic level of 40 microg x 1.5 L(-1) x day(-1). Of all wells, 44.2% had arsenic levels >50 microg/L. Multiple-adjusted odds ratios of urinary arsenic level >40 microg x 1.5 L(-1) x day(-1) were 8.90 (95% CI: 3.31-23.93) for the arsenic level in well water of 11-50 microg/L, and 53.07 (11.91-236.46) for that of 51-332 microg/L, compared with < or =10 microg/L. When the Bangladeshi standard arsenic level in drinking water of 50 microg/L was used, the sensitivity in detecting subjects with a urinary arsenic level >40 microg x 1.5 L(-1) x day(-1) was 50%, although when the World Health Organization (WHO) guideline value of 10 microg/L was used, it was 76.3%., Conclusions: Green marked wells, which the Bangladesh government regards as safe, are not always safe. The mitigation programs should use the WHO guideline arsenic level to determine the safety of well water for drinking.

A new clinical syndrome with prominent cerebellar symptoms in patients living in Kamisu City, Ibaraki Prefecture, Japan, is described. Since the patients ingested drinking water containing diphenylarsinic acid (DPA), a stable degradation product of both diphenylcyanoarsine and diphenylchloroarsine, which were developed for use as chemical weapons and cause severe vomiting and sneezing, DPA was suspected of being responsible for the clinical syndrome. The purpose of the present study was to elucidate prominent cerebellar symptoms due to DPA. The aim of the study was to determine if single (15 mg/kg) or continuous (5 mg/kg/day for 5 weeks) oral administration of DPA to ICR-strain mice induced oxidative and/or nitrosative stress in their brain. Significantly positive staining with malondialdehyde (MDA) and 3-nitrotyrosine (3-NT) was observed in the cerebellar Purkinje cells by repeated administration (5 mg/kg/day) with DPA for 5 weeks that led to the cerebellar symptoms from a behavioral pharmacology standpoint and by single administration of DPA (15 mg/kg). Furthermore, it is possible that the production of 3-NT was not caused by peroxynitrite formation. The present results suggest the possibility that arsenic-associated novel active species may be a factor underlying the oxidative and nitrosative stress in Purkinje cells due to exposure to DPA, and that the damage may lead to the cerebellar symptoms.

To solve malodorous odor problems by ammonia emission in composting of cattle dung wastes, we developed an alternative composting method consisting of a hyperthrmophilic pre-treatment reactor (HTPRT) (first step) combined with a general windrow post-treatment system (WPOT) (second step). In this study, physicochemical and microbiological differences in compost materials during the HTPRT-WPOT process and a simple windrow composing process (SWC) were investigated. The HTPRT-WPOT process removed excess ammonia in the compost materials by physical ammonia stripping, and controlled the malodorous ammonia emission. The organic matter evolution index showed that the HTPRT-WPOT process also contributed to accelerate formation of humic acids in composting. Quantitative real-time PCR analyses using Bacterial-, Archaeal- and fungal-protozoan-specific primer sets showed that small subunit ribosomal RNA (SSU rRNA) gene copy numbers differed much between composting materials of these two processes. Particularly, the SSU rRNA gene copy of eukaryotic microbes (fungi-protozoa) in the HTPRT-WPOT process was much higher than in the SWC process. From these results, we conclude that the HTPRT-WPOT process has great advantages for the control of malodorous odor problems caused by ammonia emission, and for high rate of composting evaluated by the humification rate and microbial characterization of the composting materials.

Background: Previous studies have demonstrated that atheroembolism during percutaneous coronary intervention is associated with myocardial damage. The purpose of this study is to investigate the clinical and angiographic characteristics related to removable plaque elements in patients undergoing thrombectomy for myocardial infarction., Methods: Eighty consecutive lesions in 80 patients (M/F=58/22, age 65.5+/-11.6 years) with myocardial infarction who underwent thrombectomy (TVAC system, Nipro, Osaka, Japan) prior to mechanical dilatation (balloon angioplasty and/or stent implantation) were investigated. Visible debris was collected and plaque elements (cholesterol clefts and/or foamy cells) were investigated pathologically. Baseline angiographic characteristics [baseline thrombolysis in myocardial infarction (TIMI) grade, culprit lesion, haziness, lesion length, ostium, bifurcation, calcification, eccentricity, thrombus, and multivessel] were analyzed, and predictive angiographic and clinical factors for plaque elements were investigated., Results: There were no complications related to thrombectomy. Final TIMI grade 3 and blush grade 2 or 3 were achieved in 75 (94%) and 66 (83%) patients, respectively. Visible debris specimens were obtained in 49 (61%) patients. Histological plaque elements (cholesterol clefts and/or foamy cells) were observed in 27 out of 49 patients with debris specimens. There was no significant difference in the clinical characteristics between the groups of patients with (group P) and without (group NP) plaque elements. Aspirated plaque elements were more frequently observed in discrete and eccentric lesions (group P vs. group NP: discreteness, 52% vs. 28%, P<.05; eccentricity, 67% vs. 36%, P<.05)., Conclusions: This study demonstrated the clinical characteristics associated with removable plaque components in patients with myocardial infarction undergoing thrombectomy by means of the TVAC system. Discreteness and eccentricity were more frequently observed in lesions with removable plaque elements.

An analytical method using a combination of solid-phase extraction (SPE) and gas chromatography with a flame thermionic detector (GC/FTD) was developed for determination of N-methyl-2-pyrrolidone (NMP), N-methylsuccinimide (MSI), and 2-hydroxy-N-methylsuccinimide (2-HMSI) in human urine. The SPE cartridge of poly(divinylbenzene/hydroxymethacrylate) used was directly loaded with urine sample, followed by elution with methyl isobutyl ketone (MIBK) and subsequent centrifugation, and the supernatant was injected into the capillary GC using a DB1701. This method allowed efficient separation of NMP, MSI, and 2-HMSI, which were nearly free of interference by other GC peaks arising from urine. Recoveries of NMP, MSI, and 2-HMSI from the SPE cartridge were about 98, 101, and 67%, respectively, with limits of detection of 0.04, 0.02, and 0.06 mg/L, respectively, which met the regulatory requirements. The present method was used for assay in biological monitoring of workers exposed to NMP in their occupational environment.

Formaldehyde is a carcinogen to which humans are exposed daily, but few methods are available to quantify formaldehyde in biological samples. We developed a simple, sensitive and rapid technique for the quantification of formaldehyde in urine by derivatization with O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine, using a headspace sampler coupled to a gas chromatograph equipped with an electron capture detector. The detection limit was 1.08 microg/L. The overall recovery of formaldehyde spiked in urine was 99%. The concentration of formaldehyde in urine obtained from healthy volunteers ranged from 56.85 to 144.57 microg/L. This method can be used successfully to measure formaldehyde in urine.

Although the Internet has been widely used in Japan, the quality of information on mental health-related issues has not been evaluated so far. The purpose of the present study was to investigate the current status of Japanese websites that deal with mental health, mental disorders, and their associated matters. Using a search engine, Google, 37 websites were identified that exclusively contain information on mental health or mental disorders. The characteristics of the sites were then examined, along with variety of mental disorders mentioned, and quality of contents for each of the 37 websites. More than half of the websites were set up, at least in part, for commercial purposes and only 27% of sites were owned by professionals. Mood disorder, panic disorder, and schizophrenia were the three most commonly referred disorders on websites. Aside from some exceptions, the quality of information was inadequate, especially that regarding treatment. Most of the websites on mental health and mental disorders examined in the present study have scope for improvement. The challenge is to establish a system to evaluate the sites and to motivate each webmaster to improve the sites.

Overexpression of a mercuric ion binding protein, MerP, from the mercury resistance operon genes of Gram-positive bacterial strain Bacillus megaterium MB1 and from Gram-negative bacterial strain Pseudomonas aeruginosa K-62 was found to enhance the mercury resistance level of Escherichia coli host cells, even though they share only 27.3% identity. Immunoblot analysis showed that MerP (BMerP) from Bacillus could be expressed on the membrane fraction of E. coli cells. Treated with 10 microM Hg2+, a recombinant strain harboring the BMerP gene significantly improved, showing a 27% increase in mercuric ion adsorption capacity, 16% better than that of a Pseudomonas merP gene (PMerP)-harboring strain. While multiple heavy metals co-existed, the mercuric ion adsorption capacity of the BMerP-harboring E. coli was not affected while that of the PMerP-harboring strain decreased. These results suggest that BMerP can act as a bio-adsorbent compartmentalizing the toxic mercuric ion on the cell membrane and enhancing resistance.

The toxicity and carcinogenicity of arsenic depend on its species. Individuals living in Japan consume much seafood that contains high levels of organoarsenics. Speciation analysis of urinary arsenic is required to clarify the health risks of arsenic intake. There has been no report of urinary arsenic analysis in Japan using high performance liquid chromatography with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). We performed speciation analysis of urinary arsenic for 210 Japanese male subjects without occupational exposure using HPLC-ICP-MS. The median values of urinary arsenics were as follows: sodium arsenite (AsIII), 3.5; sodium arsenate (AsV), 0.1; monomethylarsonic acid (MMA), 3.1; dimethylarsinic acid (DMA), 42.6; arsenobetaine (AsBe), 61.3; arsenocholine, trimethylarsine oxide, and unidentified arsenics (others), 5.2; and total arsenic (total As), 141.3 microgAs/l. The median creatinine-adjusted values were as follows: AsIII, 3.0; AsV, 0.1; MMA, 2.6; DMA, 35.9; AsBe, 52.1; others 3.5; and total As, 114.9 microgAs/g creatinine. Our findings indicate that DMA and AsBe levels in Japan are much higher than those found in Italian and American studies. It appears that the high levels of DMA and AsBe observed in Japan may be due in part to seafood intake. ACGIH and DFG set the BEI and BAT values for occupational arsenic exposure as 35 microgAs/l and 50 microgAs/l, respectively, using the sum of inorganic arsenic (iAs), MMA, and DMA. In the general Japanese population, the sums of these were above 50 microgAs/l in 115 (55%) samples. We therefore recommend excluding DMA concentration in monitoring of iAs exposure.

Background: The aim of this study was to measure the patient entrance skin dose (ESD) during percutaneous coronary intervention (PCI) for chronic total occlusion (CTO), and assess the factors that affect it., Methods and Results: Radiosensitive indicators were used to measure ESDs during 23 procedures. Multiple regression analysis identified the strength of the linear relationship of the dependent variable (the natural logarithm of the maximum ESD) with the set of multiple independent variables (the natural logarithm of both the patient and angiographic data). The methods for estimating the maximum ESD and the estimated ESDs were compared with the actual ESDs. The average maximum ESD for all the patients was 2.7+/-1.5 Gy (median: 2.6 Gy). The natural logarithm of the maximum ESD correlated well with the natural logarithm of body mass index (BMI; p=0.0112), total fluoroscopic time (TFT; p=0.0002), and Frame Fixation Rate (p=0.0014). For the higher Frame Fixation Rate group, there were significant correlations between the BMI2 x TFT value and maximum ESD (r=0.972, p<0.0001), and the TFT and maximum ESD (r=0.968, p<0.0001). There were no significant correlations for the lower Frame Fixation Rate group., Conclusions: In PCI for CTO, the BMI2 x TFT value or TFT is a good predictor of radiation skin injury risk, when the beam angulation is not changed frequently. It is important to change the beam angulation to control ESD during a prolonged procedure.

Brugada syndrome is an arrhythmogenetic disease characterized by electrocardiographic ST segment elevation in right precordial leads, which is called "coved type", and an increased risk of sudden death as the result of ventricular fibrillation. We presented a case of Brugada syndrome with a convulsive seizure, during administration of a tricyclic antidepressant for the treatment of a depressive state. A 43-year-old man with bipolar II disorder and obsessive-compulsive disorder was admitted to our hospital for the treatment of a depressive state. There was no medical history of heart failure. Nortriptyline was effective for his depressive as well as for obsessive symptoms. During the treatment, however, he presented a convulsive syncope. Electrocardiography (ECG) showed "coved type" ST segment elevation, and the patient consulted a cardiologist. Electrophysiological study revealed Brugada syndrome, and an implantable cardioverter defibrillator was placed. An overdose of antidepressants has been reported to produce a Brugada-type ECG because of its Na channel antagonism. However, in the present case, the abnormal ECG findings occurred following a usual dosage of nortriptyline. Thus, it is suggested that Brugada syndrome is related to a susceptibility to antidepressants in the present case. Every psychiatrist managing antidepressant therapy should be aware of Brugada syndrome and this ECG pattern, which may be a marker of sudden death.

As an initial step to understand the mobile nature of class II mercury resistance transposon TnMERI1, the effect of the recA gene on translocation of mini-TnMERI1 was evaluated. A higher transposition frequency in the LE392 strain (2.4+/-1.2x10(-5)) than in the recA-deficient DH1 strain (1.2+/-0.8x10(-6)) indicated participation of the recA gene in mini-TnMERI1 transposition. Introduction of the recA gene into the DH1 strain complemented the transposition frequency at the same level as in LE392 and confirmed participation of the recA gene in transposition. However, treatment of cells by stress agents, including irradiation of up to 3000 Jm(-2) UV doses, did not alter the transposition frequency and suggested independence of RecA from the SOS stress response. Further analysis of transconjugants indicated participation of RecA in the resolution of the cointegrate structure of the transposon. These results suggested that RecA is a constitutive cellular factor that increases translocation of mini-TnMERI1 and may participate in dissemination of TnMERI1-like transposons.

In order to identify potential genotoxicant(s) in recycled paperboard, samples were fractionated using multiple liquid/liquid extraction, and gel permeation chromatography, and analysed by gas chromatography/mass spectrometry and liquid chromatography/mass spectrometry. The rec-assay was used as an indicator of genotoxicity. Genotoxicants in the recycled paperboard were identified as dehydroabietic acid (DHA) and abietic acid (AA). DHA and AA were detected in two out of five virgin products, and in all seven recycled products for food-contact use. Total amounts of DHA and AA were 240 and 990 microg/g in the virgin products and 200-990 microg/g in the recycled products. A good correlation was observed in the total amount of DHA and AA content determined in paper products and DNA-damaging activity. Moreover, genotoxic effects in paper products showed a good match with standard compounds, indicating that the genotoxic effects of these paper products was mostly attributable to DHA and AA.

The adverse health effects caused by indoor air pollution are termed "sick building syndrome". We report such a patient whose symptoms appeared in the workplace. A 36-year-old female office worker developed nausea and headache during working hours in a refurbished office. After eight months of seeking help at other clinics or hospitals without improvement, she was referred to our hospital. At that time she reacted to the smells of various chemicals outside of the office building. Biochemical findings were all within normal ranges. Specific IgE antibody to cedar pollen was positive and the ratio of TH1/TH2 was 4.5. In the Eye Tracking Test (ETT), vertical eye movement was saccadic. Her anxiety level was very high according to the State-Trait Anxiety Inventory (STAI) questionnaire. Subjective symptoms, ETT findings and anxiety levels on STAI gradually improved during two years of follow-up. One year after the onset of her illness, the formaldehyde concentrations in the building air ranged from 0.017-0.053 ppm. Even though relatively low, chemical exposure from building materials such as formaldehyde induced a range of symptoms. Also, lack of recognition by superiors and doctors that sick building syndrome might have been the source of her illness coupled with her high state of anxiety may have exacerbated her symptoms and led to the onset of multiple chemical sensitivity. Thus psychosocial factors may contribute to sick building syndrome in the workplace.

Dimethylarsinic acid [DMA, (CH(3))(2)AsO(OH)] causes cancer in the urinary bladder of rats. However, its mechanism of cancer or the ultimate carcinogenic form is not yet known. Rats administered dimethylarsinic acid excrete three unknown arsenic compounds (termed M-1, M-2, and M-3) in urine or feces, and these compounds are presumed to be produced by intestinal bacteria. Escherichia coli A3-6 isolated from a rat yielded two unknown arsenic compounds (M-2 and M-3) from dimethylarsinic acid and M-1 from trimethylarsine oxide (TMAO) in the presence of cysteine (Cys). Contents of M-2 and M-3 varied with cysteine concentration. The cytotoxicity and genotoxicity of the bacteria-free solution of dimethylarsinic acid or trimethylarsine oxide metabolized by E. coli A3-6 were studied using V79 cells. Dimethylarsinic acid (1 mM) metabolized by E. coli A3-6 in the presence of cysteine (1 mM) was highly cytotoxic (50% survival reduction concentration; 2.1 microM As) in V79 cells, and the toxic substance appeared to be M-2. The metabolite solution (at 2.5-10 microM total As) induced c-mitosis and tetraploids, and caused mitotic arrest, since it increased mitotic cells at the cytotoxic dose. The metabolite solution also significantly increased sister chromatid exchange (SCE) and chromosomal aberrations, most of which were chromatid gaps and chromatid breaks. A3-6 converted 96.1% of trimethylarsine oxide to M-1 in the presence of cysteine. This metabolite solution did not exhibit cytotoxicity or genotoxicity. The reported M-2 concentration in urine of rats administered levels of DMA via drinking water known to cause bladder tumors was sufficient to exhibit cytotoxic and genotoxic effects in urinary bladder. Thus, we hypothesize that intestinal bacteria play an important role in carcinogenicity of dimethylarsinic acid.

This study presents the research on the chemical analysis and genotoxicity of 28 virgin/recycled paper products in food-contact use. In the chemical analysis, paper products were extracted by reflux with ethanol, and analyzed by gas chromatography/mass spectrometry. 4,4'-bis(dimethylamino)benzophenone (Michler's ketone: MK), 4,4'-bis(diethylamino)benzophenone (DEAB), 4-(dimethylamino)benzophenone (DMAB) and bisphenol A (BPA) were found characteristically in recycled products. Seventy-five percent of the recycled paper products contained MK (1.7-12 microg/g), 67% contained DEAB (0.64-10 micro g/g), 33% contained DMAB (0.68-0.9 microg/g) and 67% contained BPA (0.19-26 microg/g). Although, BPA was also detected in virgin paper products, the detection levels in the recycled products were ten or more times higher than those in the virgin products. The genotoxicity of paper and paperboard extracts and compounds found in them were investigated by Rec-assay and comet assay. Of the 28 products tested by Rec-assay using Bacillus subtilis, 13 possessed DNA-damaging activity. More recycled than virgin products (75% against 25%) exhibited such activity, which, of the compounds, was observed in BPA, 1,2-benzisothiazoline-3-one (BIT), 2-(thiocyanomethylthio)benzothiazole, 2,4,5,6-tetrachloro-isophthalonitrile, 2,4,6-trichlorophenol (TCP), and pentachlorophenol. The critical toxicant in one virgin paper product was concluded to be BIT. Eight samples with DNA-damaging activity were also tested by comet assay using HL-60 cells; six induced comet cells significantly (five times or higher than the control) without a decrease of viable cells. TCP, BZ, DEAB, and BIT also caused a slight increase in comet cells. In conclusion, we showed that most recycled paper products contain chemicals such as MK, DEAB, DMAB, and BPA, and possess genotoxicity. However, the levels of the chemicals in the recycled products could not explain their genotoxic effects.

Although numerous epidemiological studies have indicated that human arsenic exposure is associated with increased incidences of bladder, liver, skin, and lung cancers, limited attempts have been made to understand mechanisms of carcinogenicity using animal models. Dimethylarsinic acid (DMA), an organic arsenic compound, is a major metabolite of ingested inorganic arsenics in mammals. Recent in vitro studies have proven DMA to be a potent clastogenic agent, capable of inducing DNA damage including double strand breaks and cross-link formation. In our attempts to clarify DMA carcinogenicity, we have recently shown carcinogenic effects of DMA and its related metabolites using various experimental protocols in rats and mice: (1) a multi-organ promotion bioassay in rats; (2) a two-stage promotion bioassay by DMA of rat urinary bladder and liver carcinogenesis; (3) a 2-year carcinogenicity test of DMA in rats; (4) studies on the effects of DMA on lung carcinogenesis in rats; (5) promotion of skin carcinogenesis by DMA in keratin (K6)/ornithine decarboxylase (ODC) transgenic mice; (6) carcinogenicity of DMA in p53(+/-) knockout and Mmh/8-OXOG-DNA glycolase (OGG1) mutant mice; (7) promoting effects of DMA and related organic arsenicals in rat liver; (8) promoting effects of DMA and related organic arsenicals in a rat multi-organ carcinogenesis test; and (9) 2-year carcinogenicity tests of monomethylarsonic acid (MMA) and trimethylarsine oxide (TMAO) in rats. The results revealed that the adverse effects of arsenic occurred either by promoting and initiating carcinogenesis. These data, as covered in the present review, suggest that several mechanisms may be involved in arsenic carcinogenesis.

Fifty-six mercury-resistant (Hg(R)) Bacillus strains were isolated from natural environments at various sites of the world. Southern hybridisation and polymerase chain reaction (PCR) analysis showed that 21 of the 56 isolates have closely related or identical mer operons to that of Bacillus megaterium MB1. These 21 isolates displayed a broad-spectrum mercury resistance and volatilised Hg(0). PCR amplification with a single primer and restriction fragment length polymorphism analysis showed that these 21 isolates had TnMERI1-like class II transposons. These transposons can be classified into Tn5084, Tn5085, or TnMERI1. From these results, at least three types of class II mercury resistance transposons exist in Hg(R)Bacillus and these transposons may contribute the worldwide distribution and horizontal dissemination of the mer operons among Bacillus strains in natural environments.

A method for the determination of trivalent arsenicals in urine was examined. Trivalent arsenicals, extracted as complexes with diethylammonium diethyldithiocarbamate (DDDC) into carbon tetrachloride, were determined by liquid chromatography-inductively coupled plasma mass spectrometry (LC-ICP-MS). The trivalent methylated arsenicals monomethylarsonous acid (MMA(III)), dimethylarsinous acid (DMA(III)), and trimethylarsine (TMA) were detected in urine of rats that had received dimethylarsinic acid (DMA(V)) or monomethylarsonic acid (MMA(V)) at concentration of 200 microg ml(-1) in drinking water for 24 weeks. This method is the first to permit quantification of trivalent methylated arsenicals in urine without significant changes in concentration during storage or pretreatment.

The carcinogenicity of monomethylarsonic acid (MMA(V)), a major metabolite of inorganic arsenics in human and experimental animals, was investigated in male Fischer 344 rats. A total of 129 rats at 10 weeks of age were randomly divided into three groups and received drinking water containing MMA(V) at doses of 0 (Control), 50, and 200 ppm ad libitum for 104 weeks. No significant differences were found between the control and the MMA(V)-treated groups regarding clinical signs, mortality, hematological, and serum biochemistry findings. Quantitative analysis of glutathione S-transferase placental form (GST-P) positive foci in liver revealed a significant increase of numbers and areas in the 200 ppm MMA(V)-treated group. In the urinary bladder MMA(V) induced simple hyperplasia and significantly elevated the proliferating cell nuclear antigen (PCNA)-positive index in the urothelium. A variety of tumors developed in rats of all groups, including the controls, but all were histologically similar to those known to occur spontaneously in F344 rats and there were no significant differences among the groups. Thus, it could be concluded that, under the present experimental conditions, MMA(V) induced lesions in the liver and urinary bladder, but did not cause tumor development in male F344 rats even after 2 years exposure.

Arsenic is a notorious environmental toxicant known to be carcinogenic for the skin, lung and urinary bladder in human beings. The carcinogenicity of trimethylarsine oxide (TMAO), one organic metabolite of inorganic arsenics in humans and experimental animals, was investigated here in male Fischer 344 rats in a 2-year carcinogenicity test. TMAO was administered to a total of 129 male rats ad libitum at concentrations of 0 (Control), 50 or 200 p.p.m. in the drinking water. In animals that died or were killed from the 87th week until the end of 104th week, incidences of hepatocellular adenomas were 14.3, 23.8 and 35.6% in the 0, 50 and 200 p.p.m.-treated groups, respectively; the multiplicities were 0.21, 0.33 and 0.53. Both were significantly increased in the 200 p.p.m.-treated group. While a variety of other tumors developed in various organs, they were present in all groups, including the controls, and were histologically diagnosed as those known to occur spontaneously in F344 rats. To test the contribution of reactive oxygen species (ROS) to TMAO tumorigenicity in the liver, 8-hydroxydeoxyguanosine (8-OHdG) formation was assessed by high performance liquid chromatography. The 8-OHdG values for the 200 p.p.m. TMAO group were significantly higher than those for the control group. Furthermore, as assessed by the proliferating cell nuclear antigen index, cell proliferation in the normally appearing parenchyma was elevated by the TMAO treatment. These results indicate that TMAO exerts liver tumorigenicity with possible mechanistic roles for oxidative DNA damage and enhanced cell proliferation.

Thirty mercury-resistant (Hg R) Bacillus strains were isolated from mercury-polluted sediment of Minamata Bay, Japan. Mercury resistance phenotypes were classified into broad-spectrum (resistant to inorganic Hg(2+) and organomercurials) and narrow-spectrum (resistant to inorganic Hg(2+) and sensitive to organomercurials) groups. Polymerase chain reaction (PCR) product sizes and the restriction nuclease site maps of mer operon regions from all broad-spectrum Hg R Bacillus were identical to that of Bacillus megaterium MB1. On the other hand, the PCR products of the targeted merP (extracellular mercury-binding protein gene) and merA (intracellular mercury reductase protein gene) regions from the narrow-spectrum Hg R Bacillus were generally smaller than those of the B. megaterium MB1 mer determinant. Diversity of gene structure configurations was also observed by restriction fragment length polymorphism (RFLP) profiles of the merA PCR products from the narrow-spectrum Hg R Bacillus. The genetic diversity of narrow-spectrum mer operons was greater than that of broad-spectrum ones.

There is abundant epidemiological evidence that arsenic is an environmental carcinogen related to human cancers of the skin, lung, liver and urinary bladder, in particular. Dimethylarsinic acid (DMA) has also been reported to act as a carcinogen/or a promoter in rat models. To elucidate molecular mechanisms, we conducted an 18 month carcinogenicity study of DMA in p53 heterozygous (+/-) knockout mice, which are susceptible to early spontaneous development of various types of tumors, and wild-type (+/+) C57BL/6J mice. Totals of 88-90 males, 7-8 weeks of age, were divided into three groups each administered 0, 50 or 200 p.p.m. DMA in their drinking water for 18 months. Mice that were found moribund or died before the end of the study were autopsied to evaluate the tumor induction levels, as well as those killed at the end. Both p53(+/-) knockout and wild-type mice demonstrated spontaneous tumor development, but lesions were more prevalent in the knockout case. Carcinogenic effect of DMA was evident by significant early induction of tumors in both treated p53(+/-) knockout and wild-type mice, significant increase of the tumor multiplicity in 200 p.p.m.-treated p53(+/-) knockout mice, and by significant increase in the incidence and multiplicity of tumors (malignant lymphomas) in the treated wild-type mice. By the end of 80 weeks, tumor induction, particularly malignant lymphomas and sarcomas, were similar in treated and control p53(+/-) knockout mice. No evidence for organ-tumor specificity of DMA was obtained. Molecular analysis using PCR-SSCP techniques revealed no p53 mutations in lymphomas from either p53(+/-) knockout or wild-type mice. In conclusion, DMA primarily exerted its carcinogenic effect on spontaneous development of tumors with both of the animal genotypes investigated here.

The broad-spectrum mercury resistance transposon, TnMERI1, of Bacillus megaterium strain MB1, contains three proposed operator/promoter (O/P) transcriptional start sites and two regulatory genes (merR1 and merR2). A series of luciferase (lux)-based transcriptional fusion plasmids were studied in Escherichia coli to show that both merR1 and merR2 gene products repressed transcription from O/PmerB3, O/PmerR1, and O/PmerR2 under uninduced conditions. Derepression occurred when the merR1 gene was present and Hg(2+) functioned as an inducer. In the presence of organomercurial compounds, basal transcription of merB3 was needed to produce inorganic Hg(2+) as the inducer of expression regulated by MerR1 at O/PmerB3. The presence of merR2 repressed transcription from all three O/Pmer sites under both non-induced conditions and when inorganic Hg(2+) or organomercurials were added. These results show that MerR1 functions as a repressor in the absence of Hg(2+) and as an activator in the presence of Hg(2+), while MerR2 functions as a repressor.

Arsenic is a well-documented human carcinogen, and contamination with this heavy metal is of global concern, presenting a major issue in environmental health. However, the mechanism by which arsenic induces cancer is unknown, in large part due to the lack of an appropriate animal model. In the present set of experiments, we focused on dimethylarsinic acid (DMA), a major metabolite of arsenic in most mammals including humans. We provide, for the first time, the full data, including detailed pathology, of the carcinogenicity of DMA in male F344 rats in a 2-year bioassay, along with the first assessment of the genetic alteration patterns in the induced rat urinary bladder tumors. Additionally, to test the hypothesis that reactive oxygen species (ROS) may play a role in DMA carcinogenesis, 8-hydroxy-2'-deoxyguanosine (8-OHdG) formation in urinary bladder was examined. In experiment 1, a total of 144 male F344 rats at 10 weeks of age were randomly divided into four groups that received DMA at concentrations of 0, 12.5, 50 and 200 p.p.m. in the drinking water, respectively, for 104 weeks. From weeks 97-104, urinary bladder tumors were observed in 8 of 31 and 12 of 31 rats in groups treated with 50 and 200 p.p.m. DMA, respectively, and the preneoplastic lesion, papillary or nodular hyperplasias (PN hyperplasia), was noted in 12 and 14 rats, respectively. DMA treatment did not cause tumors in other organs and no urinary bladder tumors or preneoplastic lesions were evident in the 0 and 12.5 p.p.m.-treated groups. Urinary levels of arsenicals increased significantly in a dose-responsive manner except for arsenobetaine (AsBe). DMA and trimethylarsine oxide (TMAO) were the major compounds detected in the urine, with small amounts of monomethylarsonic acid (MMA) and tetramethylarsonium (TeMa) also detected. Significantly increased 5-bromo-2'-deoxyuridine (BrdU) labeling indices were observed in the morphologically normal epithelium of the groups treated with 50 and 200 p.p.m. DMA. Mutation analysis showed that DMA-induced rat urinary bladder tumors had a low rate of H-ras mutations (2 of 20, 10%). No alterations of the p53, K-ras or beta-catenin genes were detected. Only one TCC (6%) demonstrated nuclear accumulation of p53 protein by immunohistochemistry. In 16 of 18 (89%) of the TTCs and 3 of 4 (75%) of the papillomas, decreased p27(kip1) expression could be demonstrated. Cyclin D1 overexpression was observed in 26 of 47 (55%) PN hyperplasias, 3 of 4 (75%) papillomas, and 10 of 18 (56%) TCCs. As a molecular marker of oxidative stress, increased COX-2 expression was noted in 17 of 18 (94%) TCCs, 4 of 4 (100%) papillomas, and 39 of 47 (83%) PN hyperplasias. In experiment 2, 8-OHdG formation in urinary bladder was significantly increased after treatment with 200 p.p.m. DMA in the drinking water for 2 weeks compared with the controls. The studies demonstrated DMA to be a carcinogen for the rat urinary bladder and suggested that DMA exposure may be relevant to the carcinogenic risk of inorganic arsenic in humans. Diverse genetic alterations observed in DMA-induced urinary bladder tumors imply that multiple genes are involved in stages of DMA-induced tumor development. Furthermore, generation of ROS is likely to play an important role in the early stages of DMA carcinogenesis.

Objective: To investigate the relationship between blood pressure at 4 min after exercise using a Master's two-step and the risk for hypertension., Design: Prospective cohort study., Setting: Work site in Osaka, Japan., Participants: A total of 6557 Japanese men, aged 35-63 years with systolic blood pressure (SBP) < 140 mmHg and diastolic blood pressure (DBP) < 90 mmHg, and no history of hypertension or diabetes at baseline., Main Outcome Measures: Blood pressure was measured by standard techniques, using 160/95 mmHg for diagnosis of hypertension. Normotension was defined as no history of hypertension, and SBP < 130 mmHg and DBP < 85 mmHg. High normal blood pressure was defined as no history of hypertension and SBP >or= 130 and < 140 mmHg or DBP >or= 85 and < 90 mmHg., Results: During the 63 696 person-years follow-up period, we confirmed 660 cases of hypertension. SBP and DBP after exercise were associated with an increased risk for developing hypertension. The multiple-adjusted relative risk for SBP and DBP after exercise were 1.55 per 10 mmHg (confidence interval, 1.42-1.69) and 1.55 per 10 mmHg (confidence interval, 1.42-1.69), respectively. These associations were independent of resting SBP and DBP. Even after stratifying subjects according to blood pressure at rest, SBP or DBP at 4 min after exercise was associated with an increased risk for hypertension in subjects with normotension or high normal blood pressure at rest., Conclusions: The blood pressure response after exercise with a two-step was associated with an increased risk for hypertension, independently of resting blood pressures.

An Escherichia coli strain was generated by fusion of a merA-deleted broad-spectrum mer operon from Pseudomonas K-62 with a bacterial polyphosphate kinase gene (ppk) from Klebsiella aerogenes in vector pUC119. A large amount of the ppk-specified polyphosphate was identified in the mercury-induced bacterium with the fusion plasmid designated pMKB18 but not in the cells without mercury induction. These results suggest that the synthesis of polyphosphate as well as the expression of the mer genes is mercury-inducible and regulated by merR. The E. coli strain with pMKB18 was more resistant to both Hg2+ and C6H5Hg+ than its isogenic strain with cloning vector pUC119. The recombinant strain accumulated more mercury from Hg2+- and C6H5Hg+-contaminated medium. Hg2+ transported into the cytoplasm appeared to be bound by chelation with the polyphosphate produced by the recombinant cells. The transported phenylmercury was degraded to Hg2+ before the chelation since polyphosphate did not directly chelate with C6H5Hg+. These results indicate that polyphosphate is capable of reducing the cytotoxicity of the transported Hg2+ probably via chelation between polyphosphate and Hg2+.

We previously reported that detection of two specific points (the t-point and the s-point) in the relationship between pump speed and Motor Current Amplitude index (ICA) indicates the safe driving range for a continuous-flow ventricular assist device (CFVAD). During the first stage of the present experiment, the characteristic curves relating pump speed and ICA were determined by varying preload (left atrial pressure: -6 to 30 mm Hg), afterload (total circuit resistance: 890 to 3,180 dyne x sec x cm(-5)), and contractility of the left ventricle (total circuit flow: 0.5 to 2.1 L/min). These data showed that an ICA value of 0.18 was always located between the t- and s-points. During the second stage of the experiment, we developed an automatic driving program to control pump speed by maintaining ICA at 0.18. This program was able to drive the CFVAD, without exhibiting regurgitant flow or sucking, under various driving conditions in the mock circulation. Pump speed stabilized within 1 minute after varying the drive conditions. This sensorless method of driving the CFVAD by using a target ICA proved feasible and effective for safe automatic control, within our mock circulation.

To evaluate the utility of polyphosphate kinase gene (ppk)-specified polyphosphate in mercury remediation, a fusion plasmid, pMK27, with ppk from Klebsiella aerogenes and mercury transport genes, merT and merP, from Pseudomonas K-62, was constructed. The transcription and translation of ppk, merT and merP were found to be mercury-inducible. The ppk-specified polyphosphate was identified in cells preinduced by Hg2+, but not in cells without mercury induction, suggesting that the synthesis of polyphosphate is regulated by merR. The hypersensitive phenotype to Hg2+, shown by bacteria with pMRD141, which contains merT and merP, was almost completely restored to its original levels when the ppk was introduced into the plasmid, suggesting that the Hg2+-toxicity was reduced by the polyphosphate, probably via chelation formation. Bacteria with pMK27 accumulated approximately 6-fold more mercury than the bacteria with cloning vector, pUC119. These results clearly demonstrate that the polyphosphate is capable of retaining mercury in the cells without taxing the cells. Based on the results obtained in the present study, the fusion plasmid pMK27 may serve as a strategy for mercury remediation.

Formaldehyde (HCHO) is the most serious residential pollutant. In order to evaluate residential HCHO levels, two sampling methods have been recommended; one is a 30 minute sampling in a closed room, and the other is a 24 hour sampling with an ordinary lifestyle routine. The aim of this report was to clarify the difference between the HCHO levels obtained by the two sampling methods. Residential air in 58 rooms was sampled for 30 minutes by an active sampling method more than 5 hours after residents closed windows, and by a passive sampling method for 24 hours with an ordinary lifestyle routine. The HCHO concentration with the 30 minute sampling was 0.118+/-0.065 ppm (range: 0.034-0.295 ppm) and 36 rooms (62%) exceeded the Japanese guideline value of 0.08 ppm, while 5% were higher than 0.25 ppm. The HCHO concentration with the 24 hours sampling was 0.053+/-0.039 ppm (range: 0.02-0.167 ppm) and only 13 rooms (22%) exceeded 0.08 ppm. The relationship between the concentrations obtained by the two methods was linear. However, the level with the 24 hour sampling significantly reduced with prolonged window opening time, meaning that occupants made an effort to reduce the usual exposure to about 40% of the exposure in a closed room by opening windows in order to escape from irritation. Since major adverse effects of HCHO are irritation and sensitization, the occasional peak concentration must be focused. In order to evaluate residential HCHO levels, measurement in a closed room is recommended even if people are living there.

The index of motor current amplitude (ICA) has feasibility in continuous-flow ventricular assist device control. It can demonstrate the safe range of pump speed, which exists between the starting point of total assistance (t-point) and the starting point of sucking (s-point). The objective of this study was to investigate how the ICA characteristic curve changes with each condition of contractility, preload, and afterload changes. We changed preload, afterload, and contractility of closed-mock circulation and plotted the change of the ICA value against pump speed. Then the shift of ICA characteristic curve against the change of each condition was considered. When preload increased, ICA characteristic curves showed the expansion of a safe range. When afterload increased, ICA characteristic curves were shifted to the high rotation side, slightly narrowing a safe range. When contractility increased, ICA characteristic curves showed the shift of a convex above to narrowing of a safe range. As these shift patterns were observed even when the driving conditions of a circuit changed, reproducibility was checked. Understanding the feature of a shift pattern of ICA characteristic curves correctly, a possibility that change of the heart function could be predicted by change of ICA value and a possibility for a flexible control method based on ICA, according to hemodynamic state, were suggested.

Objective: To investigate the association of serum uric acid level with the risk for hypertension and Type 2 diabetes., Design: Prospective cohort study., Setting: Work site in Osaka, Japan., Participants: A total of 6,356 Japanese men, aged 35-60 years with systolic blood pressure < 140 mmHg and diastolic blood pressure < 90 mmHg, normal glucose intolerance, and no history of hypertension or diabetes at baseline., Main Outcome Measures: Blood pressure was measured by standard techniques, using 160/95 mmHg for diagnosis of hypertension. Type 2 diabetes was defined as a fasting plasma glucose level > or = 126 mg/dl or a 2 h post-loaded plasma glucose level > or = 200 mg/dl., Results: During the 61,716 person-years follow-up period, we confirmed 639 cases of hypertension and 454 cases of Type 2 diabetes. Serum uric acid level was associated with an increased risk for hypertension but not for Type 2 diabetes. After adjustment for known risk factors, including daily alcohol consumption, the serum uric acid level was associated with an increased risk for hypertension; the relative risks for hypertension were 1.00 for quintile 1 of the serum uric acid level, 1.24 [95% confidence interval (CI), 0.94-1.65] for quintile 2, 1.34 (CI, 1.03-1.76) for quintile 3, 1.76 (CI, 1.35-2.29) for quintile 4, and 2.01 (CI, 1.56-2.60) for quintile 5 (P for trend < 0.001). Even among both non-drinkers and lean subjects, serum uric acid level was associated with an increased risk for hypertension., Conclusions: Serum uric acid level was associated with an increased risk for hypertension but not for Type 2 diabetes.

In order to determine guanosine-5'-triphosphatase (GTPase) activity, we developed a simple, rapid and reliable method that utilizes capillary electrophoresis without radioisotope. Tubulin-GTPase was used for simple measurement of GTPase activity utilizing capillary electrophoresis. Tubulin, a component of microtubules, was incubated with guanosine-5'-triphosphate (GTP) in 100 mM 2-(N-morpholino) ethanesulfonic acid (MES) buffer (pH 6.5). Guanosine-5'-diphosphate (GDP) was determined as the hydrolyzed product of GTP. Guanosine-5'-monophosphate, GDP and GTP in the filtrate of the mixture were clearly separated using 10 mM MES buffer (pH 6.5) (migration time, 3.8, 5.5 and 7.2 minutes, respectively) with a fused-silica capillary column. The quantification of GDP was based on the peak area, which increased linearly with the concentration of GDP from 1 to 50 microM (r2=0.995). The peak area and migration time had good reproducibility; the intra-assay coefficient of variation (n=6) was 1.3% for peak area and 0.6% for migration time. As an application of this method, we examined the effect of dimethylarsinic acid, an effective antimitotic agent, on tubulin-GTPase. Dimethylarsinic acid inhibited tubulin-GTPase activity in a dose-dependent manner. The inhibition was not complete and the maximum decrease of the activity was about 50% at 200 microM dimethylarsinic acid. Thus, since this method is clean, simple and rapid, its application to the study of various GTPase proteins is expected to be useful.

A left ventricular assist device (LVAD) is an effective method to rescue severe heart failure. Although some require a biventricular assist, the control method for the biventricular assist device (BVAD) with a rotary pump is rarely shown. The objective of this study was to investigate the strategy for controlling BVAD with rotary pumps by in vivo studies. Using 5 piglets, we set a BVAD through a left thoracotomy and made global ischemia for 30 min by clamping the base of the ascending aorta. After unclamping, the analysis of pumping performance acted for 6 h reperfusion. We set the target flow of the LVAD and set the right ventricular assist device (RVAD) speed limit as less than when the atrial collapse occurs. To detect the ventricular collapse without any specific sensor, we calculated the index of current amplitude from motor current waveform and simultaneous mean current value. In all cases, over 6 h of observation was performed, and the RVAD was weaned almost automatically.

Urinary histamine and Ngamma-methylhistamine (1-MH), a histamine metabolite, are highly correlated with histamine in plasma. Therefore, allergic reactions can be examined by determination of histamine and 1-MH in urine. We separated histamine, 1-MH and Nalpha-methylhistamine (N-MH) by capillary electrophoresis with UV detection at 210nm, using borate buffer (pH 9) containing 100 mM SDS. The absolute detection limits were 200, 100 and 50 pg for histamine, 1-MH and N-MH, respectively. To purify histamine 1-MH and N-MH in urine, a silica cartridge was used. Recovery rates of histamine, 1-MH and N-MH in physiological saline were 90.0, 91.4 and 95.4%, respectively. We measured histamine and 1-MH levels in urine from a normal female volunteer before and after a meal, and a male bronchial asthma patient. The results showed clearly that the concentrations of histamine and its metabolite rose after eating or asthma attack. N-MH was not detected in the urine.

Background: It is not known whether physical activity is effective in reducing the risk for hypertension., Objective: To investigate the association of the duration of the walk to work and leisure-time physical activity with the risk for hypertension., Design: Prospective cohort study., Setting: Work site in Osaka, Japan., Participants: 6017 Japanese men 35 to 60 years of age with systolic blood pressure less than 140 mm Hg, diastolic blood pressure less than 90 mm Hg, normal glucose intolerance, and no history of hypertension or diabetes at baseline., Measurements: Data on physical activity were obtained by using questionnaires. Blood pressure was measured by using a standard technique; a value of at least 160/95 mm Hg was used to diagnose hypertension., Results: During 59,784 person-years of follow-up, 626 cases of hypertension were confirmed. The duration of the walk to work was associated with a reduction in the risk for incident hypertension; multivariate-adjusted relative risks were 1.00 for a walk of 10 minutes or less (reference category), 0.88 (95% CI, 0.75 to 1.04) for an 11- to 20-minute walk, and 0.71 (CI, 0.52 to 0.97) for a walk of 21 minutes or more (P for trend = 0.02). For every 26.3 men who walk more than 20 minutes to work, one case of hypertension will be prevented., Conclusions: Walking to work and other types of physical activity decreased the risk for hypertension in Japanese men. Regular exercise can prevent hypertension.

To clarify the effect of lead exposure on peripheral hemodynamics, acceleration plethysmography (APG) was performed for 48 male subjects occupationally exposed to lead (exposure group) and 43 male subjects with no history of occupational exposure to lead (control group). In the exposure group, the blood lead concentration (Pb-B) was also measured. Each APG parameter was assessed by comparing measured data with the standard aging curves. A significant negative correlation was obtained between the parameter--b/a and Pb-B. The exposure group showed significantly lower values of parameters--b/a (p < 0.01) and d/a (p < 0.05) than the control group. The parameter--b/a in the exposure group dose-dependently decreased with increases in length of working career (duration of exposure to lead) and Pb-B. The parameter--b/a significantly (p < 0.05) decreased in subjects with working careers of 5 years or more and in subjects whose Pb-B was 40 micrograms/100 ml or more. These results suggest that lead exposure affects peripheral hemodynamics as evaluated by APG.

A survey of solvent was conducted for 196 unit work areas in 95 plants in 1994 to 1996 in Hiroshima Prefecture, Japan. The survey had been repeated every 6 months (i.e., twice a year) during the 3-year period. Sampling and analysis of the solvent vapors were carried out after national protocols set by the regulation. Toluene was most frequently detected regardless of the type of solvent work (except for degreasing), whereas the second- and the third-most common solvents varied depending on the type of solvent works. Among chlorinated hydrocarbon solvents for degreasing, dichloromethane was most widely used. Solvent concentrations were generally low as none of the median concentrations exceeded corresponding Administrative Control Levels set by the regulation, either individually or even when the assumption of additiveness was applied. Among the 1176 cases analyzed, 80% of the unit work areas were evaluated as adequate (i.e., classified as Class I). Furthermore, about 57% stayed in Class I throughout the 3 years, suggesting that solvent exposure conditions were generally quite stable. In regulatory evaluation by classification, A-sampling was decisive in most cases, whereas the role of B-sampling was limited.

The metabolism of arsenic compounds in rats was studied by comparing urinary metabolites of arsenic compounds administered for 1 wk or 7 mo. Male F344/DuCrj rats were given 100 mg As/L as monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), trimethylarsine oxide (TMAO), or arsenobetaine (AsBe), or 10 mg As/L as arsenite [As(III)] via drinking water for 7 mo. Urine was collected by forced urination after 1 wk or 7 mo. Arsenic metabolites in urine were analyzed by ion chromatography with inductively coupled plasma mass spectrometry. In the case of As(III) ingestion, a small portion of all arsenic excreted in urine (about 6%) was excreted in inorganic form, while most arsenic was excreted as methylated arsenic metabolites. Following MMA treatments for 1 wk or 7 mo, the predominant products excreted were unchanged MMA and DMA accompanied by small amounts of TMAO and tetramethylarsonium (TeMA). In the case of DMA treatment the urinary compounds found were mainly the parent DMA and TMAO with minute amounts of TeMA. TMAO was methylated to TeMA to a slight extent after 1 wk and 7 mo of administration, although most TMAO was excreted in the form of unchanged TMAO. AsBe was predominantly eliminated in urine without any transformation. Two unidentified metabolites were detected in urine after 7 mo of arsenic species exposure; the amounts of these metabolites increased in the order DMA > MMA > TMAO with only small quantities of these detected in the As(III)-treated group. These results suggest that these unidentified metabolites are formed during a demethylation process, and not during methylation. Our findings indicate that long-term exposure to As(III), MMA, or DMA decreases the proportion of TMAO elimination in urine and increases that of DMA, M-1, and M-2, and that further methylation to TMAO to TeMA does occur to a slight extent following long-term exposure to arsenical compounds in rats.

We investigated the cytogenetic effects of dimethylarsinic acid (DMA), which is the major metabolite of inorganic arsenic compounds, on mouse bone marrow cells after a single intraperitoneal injection to mice. DMA increased mitotic indices significantly at 16, 24 and 48 h after injection, and prolonged the average generation time 1.5 h at the 24 h. These results suggest that DMA may cause mitotic arrest in vivo as well as in vitro. However the activity of mitotic arrest induced by DMA was much weaker than that induced by colchicine. Metaphase cells obtained after administration of DMA without colchicine pretreatment were morphologically normal except for chromosome number, which varied by stage from the prophase to the telophase in M phase as seen after administration of saline. DMA significantly induced aneuploids. The frequencies of euploids with DMA and saline treatment were 55.1 and 94.0%, respectively, and in DMA treatment hyperploids with 1 or 2 extra chromosomes were over 80% of all aneuploids. These results suggest that aneuploidy induced by DMA might be associated with carcinogenicity of arsenic.

Phthalic anhydride (PA), used in the chemical industry, binds to proteins and causes allergic reactions. It is important to study the characteristics of antibody to PA-protein. We produced specific IgG against PA-rabbit serum albumin (RSA) by administering subcutaneous injections of PA-RSA conjugate to two rabbits. Both rabbits' sera had high titers of IgG not only to PA-RSA but also to PA-human serum albumin (HSA) and HSA. In enzyme-linked immunosorbent assay (ELISA) and ELISA HSA inhibition, specific IgG to PA-HSA revealed cross-reactivity to three other phthalyl anhydride conjugates, hexahydrophthalic anhydride (HHPA)-HSA, methylhexahydrophthalic anhydride (MHHPA)-HSA, and methyltetrahydrophthalic anhydride (MTHPA)-HSA, in both sera. Titers of IgG to HHPA-HSA, MHHPA-HSA, and MTHPA-HSA were not significantly different. On affinity chromatography, highly specific IgG to PA hapten alone was purified. In the serum not binding to PA column, specific IgG to PA-HSA was significantly less than in original serum, but levels of specific IgG to other phthalyl anhydride-HSA were unchanged. Rabbits immunized with PA-RSA produced at least two types of IgG: one is to PA hapten alone and the other may be against new antigenic determinants (NADs) on HSA.