Paracoccidioidomicosis y TBC-MR en portador de VIH/VHC / Paracoccidioidomicosis and multidrug-resistant tuberculosis (TBC-MDR) in patient coinfected with HIV and hepatitis C

A case of an adult male patient diagnosed with HIV and Hepatitis C co infection is presented. He had granu-lomatuos hepatitis and blood smear positive to Paracoccidioides brasiliensis concomitant to the detection of MDR Mycobacterium tuberculosis in sputum further complicated with reactivation of cytomegalovirus (possible pancreatitis and retinitis). Difficulties in diagnostic and therapeutic approach in a patient with multiple infections are reviewed.

Reportamos el caso de un varón de 54 años portador de VHC y VIH estadio SIDA quien tuvo hepatitis granu-lomatosa y frotis de sangre positivo a Paracoccidioides brasiliensis concomitante al hallazgo de Mycobacterhim tuberculosis multiresistente en esputo, que evolucionó con reactivación de citomegalovirus (pancreatitis probable y retinitis). Se describen las dificultades diagnósticas y terapéuticas en un paciente con múltiples infecciones.

[Paracoccidioidomicosis and multidrug-resistant tuberculosis (TBC-MDR) in patient coinfected with HIV and hepatitis C]. / Paracoccidioidomicosis y TBC-MR en portador de VIH/VHC.

A case of an adult male patient diagnosed with HIV and Hepatitis C co infection is presented. He had granu-lomatuos hepatitis and blood smear positive to Paracoccidioides brasiliensis concomitant to the detection of MDR Mycobacterium tuberculosis in sputum further complicated with reactivation of cytomegalovirus (possible pancreatitis and retinitis). Difficulties in diagnostic and therapeutic approach in a patient with multiple infections are reviewed.

Polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and coplanar polychlorinated biphenyls in human milk in Osaka City, Japan.

Forty-eight human milk samples were collected from primiparous mothers in Osaka City from June 1999 to January 2000 and analyzed for polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like coplanar polychlorinated biphenyls (CoPCBs). Mean toxic equivalents (TEQs) in the milk were 13.86 pg I-TEQ/g fat or 16.50 pg World Health Organization (WHO)-TEQ/g fat for PCDDs and PCDFs; 9.87 pg WHO-TEQ/g fat for CoPCBs; and 23.74 pg TEQ/g fat using I-TEQ values of PCDDs and PCDFs or 26.36 pg TEQ/g fat using WHO-TEQ values of PCDDs and PCDFs for total PCDDs, PCDFs, and CoPCBs. The TEQ levels of these chemicals in human milk in Osaka City were in the range of levels in human milk surveyed in Japan, but the TEQ levels of PCDDs and PCDFs and total PCDDs, PCDFs, and CoPCBs from our study were slightly higher than average TEQ levels in human milk in Japan. When comparing our data with the latest data from the United States and some European countries, the TEQ levels of PCDDs and PCDFs in human milk from Osaka City were relatively high, whereas those of CoPCBs were ranked as being of intermediate level. Only TEQ values of CoPCBs in human milk were found to correlate with the increasing age of mothers and their estimated intake of seafood during the year before pregnancy. Concentrations of PCBs 105 and 118 contributed to TEQ values of CoPCBs associated with seafood intake, whereas those of PCBs 156, 157, 114, 189, 167, and 169 contributed to TEQ values of CoPCBs associated with increasing maternal age.

Effects of cysteine on the cytotoxicity of arsenic compounds.

Effects of cysteine on the cytotoxicity of arsenic compounds, such as arsenite, arsenate, methylarsonic acid (MMA), and dimethylarsinic acid (DMA), were investigated in cultured human HL-60 cells. Using adenosine triphosphate bioluminescence assay, the rank order of the mixtures of arsenicals with cysteine was: DMA > arsenite > arsenate > MMA. The IC50 of DMA with equimolar cysteine was approximately 7.7 microM, nearly two orders of magnitude lower than that of DMA alone. Apoptotic cells were examined by the TUNEL method, and cysteine was found to enhance the induction of apoptosis by arsenicals. Using LC-ICP-MS, trivalent arsenic was detected in the mixtures of arsenate, DMA, and MMA with cysteine. These results suggested that the trivalent arsenic in the mixtures of arsenicals with cysteine might account for the enhanced cytotoxicity as well as apoptosis, and that cysteine is involved in induction of the adverse effects of arsenicals in humans.

The meaning of the turning point of the index of motor current amplitude curve in controlling a continuous flow pump or evaluation of left ventricular function.

In this series, we investigated the meaning of the t-point of index of motor current amplitude (ICA) curve from a point of view of flow rate on in vitro and in vivo studies. On mock circulation loop and left ventricular assist device (LVAD)-equipped pigs, we detected the t-point and compared the pump flow at the t-point with the simultaneous cardiac output. The pump flow at the t-point showed high correlation against the simultaneous cardiac output for in vitro or in vivo study. By detection of the t-point of the ICA curve and measuring or estimating the pump flow at t-point, the cardiac output may be assessed without any sensor in various cardiac conditions.

Genotoxicity of gardenia yellow and its components.

Gardenia fruit (Gardenia jasminoides ELLIS) is widely used as a natural food colorant and as a traditional Chinese medicine for treatment of hepatic and inflammatory diseases. "Gardenia yellow" is a natural food colorant which is extracted by ethanol from gardenia fruit. The purpose of this study was to evaluate the genotoxicity of gardenia yellow. Genotoxicity of gardenia yellow and its components, crocetin, gentiobiose (a component of crocin), geniposide and genipin (formed by hydrolysis of geniposide), was studied by Ames test, rec-assay, and sister chromatid exchange (SCE) using V79 cells. Gardenia yellow and its components were found not to be mutagenic in the Salmonella reverse mutation assay. Gardenia yellow and genipin caused damage of DNA in rec-assay. Gardenia yellow induced a significant dose-dependent increase of SCE frequency (8.6 times at 1000 microg/ml as the value for the solvent control). Only genipin induced SCEs significantly among the components of gardenia yellow. Moreover, genipin induced a significant increase of tetraploids at all doses tested (95% at 8 microg/ml). Gardenia yellow preparation was analyzed by capillary electrophoresis (CE), and geniposide was detected. However, genipin was not observed. In conclusion, we have shown that genipin possesses genotoxicity. Furthermore, there were unidentified genotoxicants in gardenia yellow.

Simultaneous detection and removal of organomercurial compounds by using the genetic expression system of an organomercury lyase from the transposon Tn MERI1.

Using a newly identified organomercury lyase gene (merB3) expression system from Tn MERI1, the mercury resistance transposon first found in Gram-positive bacteria, a dual-purpose system to detect and remove organomercurial contamination was developed. A plasmid was constructed by fusing the promoterless luxAB genes as bioluminescence reporter genes downstream of the merB3 gene and its operator/promoter region. Another plasmid, encoding mer operon genes from merR1 to merA, was also constructed to generate an expression regulatory protein, MerR1, and a mercury reductase enzyme, MerA. These two plasmids were transformed into Escherichia coli cells to produce a biological system that can detect and remove environmental organomercury contamination. Organomercurial compounds, such as neurotoxic methylmercury at nanomolar levels, were detected using the biomonitoring system within a few minutes and were removed during the next few hours.

Bio-affecting mercury detection using mercury resistance gene module fused with bioluminescence reporter genes.

Bioluminescence sensor systems were developed for monitoring environmental mercury contamination. The biological mercury measurement sensor systems were constructed by DNA recombination technique. A bacterial mercury-resistant operon (meroperon) from Pseudomonas sp. K-6y4 and a bacterial bioluminescence operon (lux operon) from an ocean bacterium Vibrio fischeri were fused in avector plasmid. The resulting recombinant plasmids were cloned in Escherichia coli cells. The bioluminescence sensor systems responded to mercury chloride of 0.1 nM to 100 nM. The mercury bioluminescence sensor developed in this study can be used for monitoring of the bio-affecting mercury instead of total mercury that is measured by conventional analytical equipment. The fundamental feature of the bioluminescence sensor system is attractive for use as a monitoring system for bio-affecting environmental mercury contamination.

The index of motor current amplitude has feasibility in control for continuous flow pumps and evaluation of left ventricular function.

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.

Serum uric acid and the risk for hypertension and Type 2 diabetes in Japanese men: The Osaka Health Survey.

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.

Sampling methods and residential factors affecting formaldehyde concentration in indoor air.

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.

Evaluation of ppk-specified polyphosphate as a mercury remedial tool.

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.

Simple and rapid determination of Gtpase activity by capillary electrophoresis without radioisotope.

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.

Control strategy for biventricular assistance with mixed-flow pumps.

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.

Sensorless controlling method for a continuous flow left ventricular assist device.

We originated a novel control strategy for a continuous flow left ventricular assist device (LVAD). We examined our method by acute animal experiments to change the left ventricular (LV) contractility or LV end-diastolic pressure (LVEDP). To estimate the pump pulsatility without any specific sensor, we calculated the index of current amplitude (ICA) from motor current waveform. The ICA had a peak point (t-i point) that corresponded closely with the turning point from partial to total assistance, and a trough (s-i point) that corresponded with the beginning point of ventricular collapse. The pump flow at the t-i point (Qt-i) had no component of flow regurgitation. In the evaluation of the effects of preload LVEDP, afterload (mAoP), and contractility (max LV dp/dt), we found that preload was the only parameter that significantly influenced Qt-i. We concluded that our method could well control continuous flow LVAD by preventing reversed flow and ventricular collapse.

Determination of histamine, 1-methylhistamine and N-methylhistamine by capillary electrophoresis with micelles.

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.

Leisure-time physical activity at weekends and the risk of Type 2 diabetes mellitus in Japanese men: the Osaka Health Survey.

AIMS: To investigate association between leisure-time physical activity at weekends and the risk of developing Type 2 diabetes mellitus (DM). METHODS: Prospective examination of 6,013 Japanese men aged 35-60 years who were free of DM, impaired fasting glycaemia, or hypertension at study entry. Type 2 DM was defined by a fasting plasma glucose level > or = 7.0 mmol/l or a 2-h post-load plasma glucose level > or =11.1 mmol/l. Data on physical activity obtained from questionnaires consisted of overall leisure-time physical activity weekly and leisure-time physical activity at weekends. RESULTS: During the 59,966 person-years follow-up, 444 cases developed Type 2 DM. Regular physical exercise at least once a week was associated with a reduced risk of Type 2 DM. After adjustments for age, body mass index, daily alcohol consumption, smoking habits, blood pressure levels and a parental history of Type 2 DM, men who engaged in regular physical exercise at least once a week had a relative risk of Type 2 DM of 0.75 (95% CI, 0.61-0.93) compared with men engaging in exercise less often. Even vigorous activity only once a week at weekends was associated with a reduced risk of Type 2 DM. Men who engaged in vigorous activity at least once a week at weekends had a multiple-adjusted relative risk of Type 2 DM of 0.55 (95% CI, 0.35-0.88) compared with sedentary men. CONCLUSIONS: Regular physical exercise at least once a week and vigorous activity even only once a week at weekends are associated with a decreased risk of Type 2 DM.

Impact of cigarette smoking on the incidence of Type 2 diabetes mellitus in middle-aged Japanese men: the Osaka Health Survey.

AIMS: To assess the impact of cigarette smoking on the incidence of Type 2 diabetes mellitus (DM) in middle-aged Japanese men. METHODS: The study enrolled 6250 men aged 35-60 years and free of diabetes, impaired fasting glucose and hypertension at entry. Type 2 DM was defined by a fasting plasma glucose level > or =7.0 mmol/l or physician-diagnosed Type 2DM. RESULTS: Four hundred and fifty cases of Type 2 DM were confirmed during the 60904 person-years follow-up. After adjustment for multiple covariates, including age, body mass index, alcohol consumption, physical activity, parental history of diabetes and the level of fasting plasma glucose, total cholesterol, triglycerides, high-density lipoprotein cholesterol and haematocrit, the relative risk of Type 2 DM among current smokers compared with non-smokers was 1.47 (95% confidence interval (CI) 1.14-1.92). Men who smoked >30 cigarettes/day had a multivariate-relative risk of 1.73 (95% CI 1.20-2.48) compared with non-smokers. The number of cigarettes smoked daily and the pack-year values were positively related to the development of Type 2 DM in a dose-dependent manner (P for trends = 0.0026 and 0.001, respectively). CONCLUSIONS: A cigarette smoking habit is an independent risk factor for Type 2 DM.

Identification of three merB genes and characterization of a broad-spectrum mercury resistance module encoded by a class II transposon of Bacillus megaterium strain MB1.

The complete structure of a broad-spectrum mercury resistance module was shown by sequencing the Gram-positive bacterial transposon TnMERI1 of Bacillus megaterium MB1. The regions encoding organomercury resistance were identified. Upstream of a previously identified organomercurial lyase merB (merB1) region of TnMERI1, a second merR (merR2) and a second merB gene (merB2) were found. These genes constitute a second operon (mer operon 2) following a promoter/operator (P(merR2)) region. A third organomercurial lyase gene (merB3) was found immediately upstream of the mer operon (mer operon 1) followed by a promoter/operator (P(merB3)) region homologous to that of the mer operon 1 (P(merR1)-merR1-merE-like-merT-merP-merA). The complete genetic structure of the mercury resistance module is organized as P(merB3)-merB3-P(merR1)-merR1-merE-like-merT+ ++ -merP-merA-P(merR2)-merR2 -merB2-merB1. The subcloning analysis of these three merB genes showed distinct substrate specificity as different organomercury lyase genes.