Distribution of virulence markers among Vibrio vulnificus isolates of clinical and environmental origin and regional characteristics in Japan.

BACKGROUND: Vibrio vulnificus is an opportunistic human pathogen that is widely distributed in estuarine environments and is capable of causing necrotizing fasciitis and sepsis. In Japan, based on epidemiological research, the incidences of V. vulnificus were concentrated in Kyusyu, mainly in coastal areas of the Ariake Sea. To examine the virulence potential, various genotyping methods have recently been developed. This study aimed to investigate the distribution of virulence markers among V. vulnificus isolates of clinical and environmental origin in three coastal areas with different infection incidences and to determine whether these isolates have the siderophore encoding gene viuB. METHODOLOGY/PRINCIPAL FINDINGS: We examined the distribution of genotypes of the 16S ribosomal ribonucleic acid (rRNA) gene, vvhA, vcg, and capsular polysaccharide (CPS), and the presence of viuB in 156 isolates collected from patients and environmental samples in Japan. The environmental samples were collected from three coastal areas: the Ariake Sea, Ise & Mikawa Bay, and Karatsu Bay. The results showed disparity in the ratios of genotypes depending on the sample origins. V. vulnificus isolates obtained from patients were classified into the clinical type for all genotypes. In the environmental isolates, the ratios of the clinical type for genotypes of the 16S rRNA gene, vvhA, and vcg were in the order of the Ariake Sea>Ise & Mikawa Bay>Karatsu Bay. Meanwhile, CPS analysis showed no significant difference. Most isolates possessed viuB. CONCLUSIONS: Many V. vulnificus belonging to the clinical type existed in the Ariake Sea. Three coastal areas with different infection incidences showed distinct ratios of genotypes. This may indicate that the distribution of clinical isolates correlates with the incidence of V. vulnificus infection.

[Difficulty in left-sided double-lumen tube placement in patients undergoing surgical graft replacement of the descending aorta is predicted from tracheobronchial compression].

BACKGROUND: In surgical graft replacement of the descending aorta graft, one-lung ventilation (OLV) is required to provide adequate surgical view and to allow removal of blood from the left lung. It is best to use a double-lumen tube (DLT) to assure OLV but it is sometimes difficult to place the left-sided DLT due to thoracic aneurysm or the dissection lumen. We retrospectively investigated tracheobronchial anatomy by chest X-ray and chest computed tomography (CT) in 29 cases of descending aorta replacement to determine how best to manage difficult placement of the left-sided DLT. METHODS: From our database of 29 patients who had undergone descending aorta replacement between February 1, 2005, and December 31, 2009, we investigated the association between difficulty in placing the left-sided DLT and tracheobronchial anatomy by chest X-ray and CT. RESULTS: We could not place a left-sided DLT in 3 of 29 cases. Two of these cases were planned surgery for aortic aneurysm and the other was an emergency operation for acute aortic dissection. We could manage the two cases safely using a right-sided DLT. We compared chest X-ray and chest CT images of these 3 cases with the other 26 cases and found that compression of the tracheobronchial tree was prevalent in the cases of difficult placement of the left-sided DLT. CONCLUSIONS: We experienced difficulty in placement of the left-sided DLT in 3 of 29 cases of descending aorta replacement. We can predict difficulty of left-sided DLT placement by the presence of compression of the tracheobronchial tree on chest CT.

Sivelestat relaxes vascular smooth muscle contraction in human gastric arteries

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Sivelestat sodium hydrate (sivelestat) is a novel synthetic drug and specific inhibitor of neutrophil elastase that has been approved in Japan as a treatment for acute lung injury associated with systemic inflammatory response syndrome. It is important to determine how sivelestat affects hemodynamics and the regulatory mechanisms of vascular smooth muscle (VSM). We recently found that sivelestat relaxes porcine coronary artery VSM via selective inhibition of Ca2+ sensitization induced by a receptor agonist without affecting the normal Ca2+-induced contraction. Although sivelestat relaxes porcine artery, its effects on human artery are unknown; therefore, the purpose of the present study was to assess the effects of sivelestat on human artery. In the present study, sivelestat induced concentration-dependent (1 × 10−6 to 3 × 10−4 M) vasorelaxation in U46619 (1 nM) and sphingosylphosphorylcholine (SPC) (30 mM)-precontracted human gastric artery with or without endothelium, but sivelestat did not induce vasorelaxation in conditions of high K+ (40 mM) depolarization. Sivelestat inhibited VSM contraction by an agonist and SPC, and it did not affect Ca2+-induced normal physiologic contraction (AU)

Sivelestat relaxes vascular smooth muscle contraction in human gastric arteries.

Sivelestat sodium hydrate (sivelestat) is a novel synthetic drug and specific inhibitor of neutrophil elastase that has been approved in Japan as a treatment for acute lung injury associated with systemic inflammatory response syndrome. It is important to determine how sivelestat affects hemodynamics and the regulatory mechanisms of vascular smooth muscle (VSM). We recently found that sivelestat relaxes porcine coronary artery VSM via selective inhibition of Ca(2+) sensitization induced by a receptor agonist without affecting the normal Ca(2+)-induced contraction. Although sivelestat relaxes porcine artery, its effects on human artery are unknown; therefore, the purpose of the present study was to assess the effects of sivelestat on human artery. In the present study, sivelestat induced concentration-dependent (1 × 10(-6) to 3 × 10(-4) M) vasorelaxation in U46619 (1 nM) and sphingosylphosphorylcholine (SPC) (30 mM)-precontracted human gastric artery with or without endothelium, but sivelestat did not induce vasorelaxation in conditions of high K(+) (40 mM) depolarization. Sivelestat inhibited VSM contraction by an agonist and SPC, and it did not affect Ca(2+)-induced normal physiologic contraction.

Carbon-14 analysis in solidified product of non-metallic solid waste by a combination of alkaline fusion and gaseous CO2 trapping.

In order to establish a simple and rapid analytical method for (14)C in solidified products made from non-metallic low-level radioactive solid wastes such as concrete, mortar and glass by melting treatment, a radiochemical analysis in combination with alkaline fusion as a sample decomposition method was examined. A simulated solidified product containing (14)C, which was prepared by using nuclear reaction (14)N(n, p)(14)C with thermal neutron irradiation, was analyzed by the present method to compare with a conventional radiochemical analysis using oxidizing combustion. The reproducible and quantitative recovery of (14)C from the simulated solidified product indicates that the present method is more efficient for (14)C analysis in solidified products than the conventional method using oxidizing combustion.

High concentrations of dexmedetomidine inhibit compound action potentials in frog sciatic nerves without alpha(2) adrenoceptor activation.

BACKGROUND AND PURPOSE: Dexmedetomidine, an alpha(2)-adrenoceptor agonist, exhibits anti-nociceptive actions at the spinal cord and enhances the effect of local anaesthetics in the peripheral nervous system. Although the latter action may be attributed in part to inhibition of nerve conduction produced by dexmedetomidine, this has not been fully examined yet. EXPERIMENTAL APPROACH: We examined the effects of various adrenoceptor agonists including dexmedetomidine, and tetracaine, a local anaesthetic, on compound action potentials (CAPs) recorded from the frog sciatic nerve, using the air-gap method. KEY RESULTS: Dexmedetomidine reversibly and concentration-dependently reduced the peak amplitude of CAPs (IC(50)= 0.40 mmol x L(-1)). This action was not antagonized by two alpha(2)-adrenoceptor antagonists, yohimbine and atipamezole; the latter antagonist itself reduced CAP peak amplitude. Clonidine and oxymetazoline, two other alpha(2)-adrenoceptor agonists, also inhibited CAPs; the maximum effect of clonidine was only 20%, while oxymetazoline was less potent (IC(50)= 1.5 mmol x L(-1)) than dexmedetomidine. On the other hand, (+/-)-adrenaline, (+/-)-noradrenaline, alpha(1)-adrenoceptor agonist (-)-phenylephrine and beta-adrenoceptor agonist (-)-isoprenaline (each 1 mmol x L(-1)) had no effect on CAPs. Tetracaine reversibly reduced CAP peak amplitude (IC(50) of 0.014 mmol x L(-1)). CONCLUSIONS AND IMPLICATIONS: Dexmedetomidine reduced CAP peak amplitude without alpha(2)-adrenoceptor activation (at concentrations >1000-fold higher than those used as alpha(2) adrenoceptor agonist), with a lower potency than tetracaine. CAPs were inhibited by other alpha(2) adrenoceptor agonists, oxymetazoline and clonidine, and also an alpha(2) adrenoceptor antagonist atipamezole. Thus, some drugs acting on alpha(2) adrenoceptors are able to block nerve conduction.

Clinical features of Vibrio vulnificus infections in the coastal areas of the Ariake Sea, Japan.

Vibrio vulnificus infection can result in necrotizing fasciitis and sepsis and is associated with high mortality. Most patients infected with this microbe have liver dysfunction as an underlying disease. However, because of the sporadic nature of outbreaks and unidentified cases, extensive evaluation of clinical features and identification of factors affecting prognosis have not been performed. We retrospectively analyzed 37 cases in Japan from 1984 to 2008 to review clinical features and to identify risk factors associated with prognosis. Statistical differences between clinical features (patient's characteristics, initial clinical laboratory data, symptoms upon admission, and other risk indicators) and prognosis were analyzed by use of the chi(2) test or the Mann-Whitney U test. Multivariate logistic regression analysis was also performed to assess factors which potentially affect hospital mortality. The mortality rate was 64.9%. An underlying liver disease was observed in 91.6% of the patients. The presence of liver cirrhosis tended to be related to hospital mortality; however, statistical significance was not achieved. Advanced age, lower platelet counts, and the presence of extensive skin lesions at onset affected outcomes with statistical significance. The prognosis of this disease is poor, because septic shock and necrotizing fasciitis often develop within a few days. Early diagnosis and treatment are needed to improve the prognosis of V. vulnificus infection.

Influence of hydrofluoric acid on extraction of thorium using a commercially available extraction chromatographic resin.

The dependence of Th recovery on hydrofluoric acid (HF) concentration in nitric acid (HNO(3)) solutions (1-5 mol/dm(3)) containing 1x10(-6) mol/dm(3) of Th and various concentrations of HF and the elution behavior were studied using a commercially available UTEVA (for uranium and tetravalent actinide) resin column. Thorium recovery decreased with an increase in HF concentration in the sample solutions. The concentration of HF at which Th recovery started to decrease was approximately 1x10(-4) mol/dm(3) in 1 mol/dm(3) HNO(3) solution, approximately 1x10(-3) mol/dm(3) in 3 mol/dm(3) HNO(3) solution, and approximately 1x10(-2) mol/dm(3) in 5 mol/dm(3) HNO(3) solution. When Al(NO(3))(3) (0.2 mol/dm(3)) or Fe(NO(3))(3) (0.6 mol/dm(3)) was added as a masking agent for F(-) to the Th solution containing 1x10(-1) mol/dm(3) HF and 1 mol/dm(3) HNO(3), Th recovery improved from 1.4+/-0.3% to 95+/-5% or 93+/-3%. Effective extraction of Th using UTEVA resin was achieved by selecting the concentration of HNO(3) and/or adding masking agents such as Al(NO(3))(3) according to the concentration of HF in the sample solution.

[Rapid detection of Vibrio vulnificus using a loop-mediated isothermal amplification method].

Vibrio vulnificus is found in marine waters near the coast around the world. Infection with this gram-negative rod, via ingestion of raw seafood or via a skin wound following contact with contaminated estuarine or marine water, can cause necrotizing fasciitis and sepsis. Most of patients with Vibrio vulnificus infection have underlying liver dysfunction or diabetes mellitus. Due to the high mortality and short latent periods, control of this infection depends on early identification of the bacterial species and prompt initiation of intensive care. Accordingly, the development of a technique that can identify this microbe quickly and accurately is of great importance. Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method to detect specific genes with rapidity and high sensitivity. In this study, we developed LAMP for the detection of Vibrio vulnificus. Using 28 Vibrio vulnificus strains and 53 other bacterial strains, we confirmed the high specificity of this method. Moreover, our LAMP method also showed high sensitivity, with a minimum detection level of one colony-forming unit per test. Furthermore, we developed simplified and conventional pretreatments for the method using experimental animal models. All of these attempts have lod to our non being able to detect Vibrio vulnificus within 1 hour.

Sivelestat relaxes porcine coronary artery via inhibition of Ca2+ sensitization induced by a receptor agonist.

Sivelestat sodium hydrate (sivelestat) is a novel synthetic drug and specific inhibitor of neutrophil elastase that has been approved in Japan as a treatment for acute lung injury associated with systemic inflammatory response syndrome. There are no reports on the effects of sivelestat on the contractile regulation of vascular smooth muscle. The purpose of the present study was to assess the effects of sivelestat on porcine coronary artery. Sivelestat induced concentration-dependent (3 x 10 to 3 x 10 M) vasorelaxation in U46619 (100 nM)-precontracted porcine coronary artery with or without endothelium. Simultaneous measurements of tension and the cytosolic Ca concentration ([Ca]i) revealed that sivelestat shifted the [Ca]i-tension curve to the right and downward during stimulation with 118 mM K and 100 nM U46619. In beta-escin-permeabilized arterial strips, sivelestat abolished GTP plus U46619-induced contractions at constant [Ca]i, whereas it had no effect on Ca-induced contractions. Thus, sivelestat relaxes porcine coronary artery smooth muscle via the selective inhibition of Ca sensitization induced by a receptor agonist, without affecting Ca-induced contraction.

Partial blood recirculation: a new trial for prolonging filter life during continuous hemodiafiltration.

Although continuous hemodiafiltration (CHDF) has been widely accepted in the management of complicated acute renal failure, the requirement for prolonged continuous systemic anticoagulation appears to be a major drawback. We herein describe the case of a patient who developed postoperative multiple organ failure and received CHDF therapy with partial blood recirculation (PBR). PBR is a mode of extracorporeal circulation used as an anticoagulation modality. The technique accelerates the blood flow rate with the goal of extending filter life, and it was performed because the filter life had been significantly shortened (10.5 +/- 5.1 h) during the CHDF process in this case. Despite increasing the dose of the anticoagulant, changing the hemofilter and changing the mode from postdilution to predilution, we did not obtain amelioration of filter life. The filter life was significantly improved (41.5 +/- 1.4 h) when we performed PBR. It is difficult to minimize the bleeding risk and maintain filter life during CHDF. Our success in prolonging the filter life during this case therefore suggests that PBR might resolve one of the main problems related to CHDF, although more study is needed to clarify the advantages of this system.

A fatal case of necrotizing fasciitis due to bacterial translocation of Klebsiella oxytoca.

We report a 73-year-old man with hepatocellular cell carcinoma who had eruptions on and severe pain in the lower leg. Within several hours, the patient's skin lesions had progressed markedly. Magnetic resonance imaging findings were consistent with necrotizing fasciitis. Klebsiella oxytoca was isolated from cultures of biopsy samples taken from the leg. The resulting DNA fingerprint pattern revealed that the enteric bacterium was the same as that obtained from the biopsy samples taken from the leg. Furthermore, a dendrogram showed that genetic proximity between samples was extremely high. These results confirmed that translocation of Klebsiella oxytoca as an enteric pathogen caused the necrotizing fasciitis in this patient.

Survival case of Vibrio vulnificus infection with multiple endocrine neoplasia type I.

A 35-year-old female with malabsorption syndrome who underwent a pancreatoduodenectomy for multiple endocrine adenomatosis 13 years prior was admitted to our hospital with diarrhea, general fatigue, high fever, and eruption in the lower legs. The patient had consumed raw shrimp a few days before onset and presented systemic inflammatory response syndrome at the time of hospitalization. Vibrio vulnificus was isolated from a blood culture performed before admission to the intensive care unit. We excised necrotizing tissue in the legs after improvement of her general condition. During the treatment process, glucose, catecholamine, and appropriate antibiotics were administered for hypoglycemia, hypotension, and high fever, respectively. The patient was discharged 107 days after contracting the disease. Of 18 septic patients with V. vulnificus infection admitted to our hospital, this was the first to develop septicemia in the absence of a previous liver dysfunction. In order to prevent this type of fatal infection, public education for immuno-compromised individuals as well as those with liver disease is essential. For early diagnosis and appropriate treatment, more effective strategies are required, such as the establishment of a network system where family physicians and emergency hospital staff could discuss information regarding high-risk patients.

A chromogenic substrate culture plate for early identification of Vibrio vulnificus and isolation of other marine Vibrios.

Vibrio vulnificus infection can result in necrotizing fasciitis and sepsis, which have short latentcy periods and high mortality rates. Thus, an easy and quick detection method is needed to improve the outcome. To distinguish V. vulnificus from other pathogens that cause necrotizing fasciitis, we developed a selective isolation culture agar plate (Chromochecker Vibrio Agar-1; CVA-1) for use in environmental monitoring and in the clinical setting. One hundred four strains of V. vulnificus, already identified biochemically, showed typical colony form and color when grown on CVA-1. Thirty-six of 51 marine bacteria samples suspected to be V. vulnificus on CVA-1 were subsequently identified as V. vulnificus by a biochemical identification system. Of 8 bacteria known to cause necrotizing fasciitis, only V. vulnificus grew on CVA-1. In addition, growth on CVA-1 allowed ready differentiation of Vibrio species. CVA-1 can be used to distinguish pathogenic Vibrios according to colony form and chromatic differences.

Application of extraction chromatography to the separation of thorium and uranium dissolved in a solution of high salt concentration.

Extraction chromatography with commercially available UTEVA resin (for uranium and tetravalent actinide) was applied for the separation of Th and U from control solutions prepared from a multi-element control solution and from sample solutions of solidified simulated waste. Thorium and U in control solutions with 1-5mol/dm(3) HNO(3) were extracted with UTEVA resin and recovered with a solution containing 0.1mol/dm(3) HNO(3) and 0.05mol/dm(3) oxalic acid to be separated from the other metallic elements. Extraction behavior of U in the sample solutions was similar to that in the control solutions, but extraction of Th was dependent on the concentration of HNO(3). Thorium was extracted from 5mol/dm(3) HNO(3) sample solutions but not from 1mol/dm(3) HNO(3) sample solutions. We conjecture that thorium fluoride formation interferes with extraction of Th. Addition of Al(NO(3))(3) and Fe(NO(3))(3), which have higher stability constant with fluoride ion than Th, does improve extractability of Th from 1mol/dm(3) HNO(3) sample solution.

Iodoform intoxication; a case report of prolonged consciousness disturbance in a patient with a high plasma iodine level.

This report describes a case in which a consciousness disturbance was associated with a high plasma iodine level and which improved after the removal of iodoform gauze that had been applied to infected wounds. A 71-year-old male with poorly controlled diabetes underwent a laminectomy for an epidural abscess. On the 6th and 8th postoperative days, he underwent debridement for fasciitis in the both arms and iodoform gauze was applied to the wounds (both arms and lumbar region) at the end of the first debridement and then changed every day. His consciousness then gradually deteriorated after the debridement. His plasma iodine level was measured and was shown to be remarkably high (6,280 microg/dl) 19 days after the laminectomy (13 days after the beginning of iodoform application). His consciousness recovered gradually with a concomitant decrease in the plasma iodine level. Three months after the removal of the iodoform he became almost alert. Caution should be exercised in using iodoform gauze because of the possibility that it could affect consciousness.

[A collective review of Vibrio vulnificus infection in Japan].

Vibrio vulnificus is found globally in marine coastal waters. Infection with this organism, via ingestion of raw shellfish or exposure to marine water, can cause necrotizing fasciitis and sepsis, which have high mortality and short latency. In Japan, many cases have been reported since 1980, mainly from hospitals in western prefectures. However, because of the sporadic nature of infection outbreaks, a thorough epidemiologic survey has not been done. We studied the epidemiological and clinical characteristics of Vibrio vulnificus infections reported in Japan from 1975 to 2005. We identified 185 cases using the medical article search engines Ichushi (Japan Medical Abstracts Society), CiNii (Citation Information by National Institute of Informatics), and PubMed over 30 years. The median age of patients was 59. The number of male patients was eight times the number of female patients; however, no significant difference in mortality was found between genders. In yearly distributions of patients, 20 cases were reported in 2001, a year in which we experienced more rainfall and a longer rainy season in Northern Kyushu. In monthly distribution, about 80% of cases were reported from July to September when sea water temperatures rise. About 40% of cases were reported in four prefectures around the Ariake Sea. The underlying disease indicated liver dysfunction in 90% of patients, but mortality was the same regardless of the infection pathway (oral ingestion or wound). Because of its rapid aggravation and high mortality rate, public education is important to prevent new cases. It is also highly recommended that patients with preexisting liver dysfunction avoid raw fish and limit exposure to marine water during the summer.

Extraction of uranium from simulated ore by the supercritical carbon dioxide fluid extraction method with nitric acid-TBP complex.

The supercritical fluid extraction (SFE) method using CO(2) as a medium with an extractant of HNO(3)-tri-n-butyl phosphate (TBP) complex was applied to extract uranium from several uranyl phosphate compounds and simulated uranium ores. An extraction method consisting of a static extraction process and a dynamic one was established, and the effects of the experimental conditions, such as pressure, temperature, and extraction time, on the extraction of uranium were ascertained. It was found that uranium could be efficiently extracted from both the uranyl phosphates and simulated ores by the SFE method using CO(2). It was thus demonstrated that the SFE method using CO(2) is useful as a pretreatment method for the analysis of uranium in ores.

[Anesthetic management of combined lung volume reduction surgery and off-pump coronary artery bypass grafting].

A 54-year-old man with severe emphysema and stenosis of coronary artery was scheduled for combined surgery of lung volume reduction and an off-pump coronary artery bypass grafting. His FEV1.0 was 600 ml and %FEV1.0 was 18%. Coronary angiography showed 99% stenosis of the left anterior descending artery. Anesthesia was induced with propofol, fentanyl and vecuronium, and was maintained with sevoflurane and continuous epidural anesthesia. In order to avoid high airway pressure, a pressure-controlled ventilation (less than 15 cmH2O) was carried out. A laryngeal mask airway was replaced with an endotracheal tube after surgery to avoid bucking during extubation, and this was removed after recovery from anesthesia successfully. No complications were observed during anesthesia. Lung volume reduction surgery after coronary reconstruction by off-pump coronary artery bypass grafting may be beneficial for patients with emphysema and ischemic heart disease.

NOS II inhibition restores attenuation of endothelium-dependent hyperpolarization in rat mesenteric artery exposed to lipopolysaccharide.

The aim of this study was to assess the effects of lipopolysaccharide (LPS) exposure on the endothelium-dependent hyperpolarization in the rat mesenteric artery using isometric tension recordings and electrophysiological studies. Mesenteric arterial rings of male Sprague-Dawley rats were incubated with LPS for 6 hours. All experiments were performed in the presence of indomethacin to inhibit the formation of vasoactive prostanoids. Contraction to phenylephrine was significantly reduced in rings incubated with LPS, which was restored in the presence of N(omega)-nitro-L-arginine methyl ester (L-NAME). L-NAME resistant relaxation to acetylcholine was attenuated in LPS-treated rings. LPS exposure hyperpolarized resting membrane potentials of arterial smooth muscle cells, which was repolarized by incubation with either L-NAME or 1400W, a selective inhibitor of nitric oxide synthase II (NOS II). Endothelium-dependent hyperpolarization to acetylcholine was attenuated in arteries incubated with LPS, while incubation with LPS and 1400W restored EDHF-mediated hyperpolarization. LPS-induced membrane potential change was mimicked by incubation with either SIN-1 or diethylamine NONOate, a donor of nitric oxide. These data suggest that LPS exposure attenuates EDHF-mediated both relaxation and hyperpolarization in the rat mesenteric artery. The possible mechanisms underlying decreased EDHF-mediated responses might be due to, at least in some part, massive nitric oxide induced by NOS II.