Rapid discrimination methods for clinical and environmental strains of Aeromonas hydrophila and A. veronii biovar sobria using the N-terminal sequence of the flaA gene and investigation of antimicrobial resistance.

Although the genus Aeromonas inhabits the natural environment, it has also been isolated from hospital patient specimens as a causative agent of Aeromonas infections. However, it is not known whether clinical strains live in the natural environment, and if these strains have acquired antimicrobial resistance. In this study, we performed the typing of flagellin A gene (flaA) of clinical and environmental strains of Aeromonas hydrophila and A. veronii biovar sobria using Polymerase Chain Reaction (PCR) assay with newly designed primers. Detection rates of the clinical and environmental flaA types of A. hydrophila were 66.7% and 88.2%, and the corresponding rates for A. veronii biovar sobria were 66.7% and 90.9%. The PCR assays could significantly discriminate between clinical and environmental strains of both species in approximately 4 h. Also, among the 63 clinical Aeromonas strains used, only one extended-spectrum ß-lactamase-producing bacteria, no plasmid-mediated quinolone resistance bacteria, and only four multidrug-resistant bacteria were detected. Therefore, the PCR assays could be useful for the rapid diagnosis of these Aeromonas infections and the monitoring of clinical strain invasion into water-related facilities and environments. Also, the frequency of drug-resistant Aeromonas in clinical isolates from Okinawa Prefecture, Japan, appeared to be low.

Differences in flaA gene sequences, swimming motility, and biofilm forming ability between clinical and environmental isolates of Aeromonas species.

The flagellin A gene (flaA) sequences, swimming motility, and biofilm forming ability were investigated in order to reveal the genetic and functional differences of flagella between clinical and environmental isolates of Aeromonas species. Twenty-eight clinical and 48 environmental strains of Aeromonas species isolated in Okinawa Prefecture of Japan were used in this study. The full-length flaA genes of these strains were sequenced and aligned, and a phylogenetic tree was constructed. In addition, swimming motility and biofilm forming ability were evaluated by conventional methods. Aeromonas veronii biovar sobria and A. hydrophila clearly divided into clinical and environmental strain clusters in the flaA phylogenetic classification, and the six and 13 specific amino acids respectively, of FlaA of both species were different in clinical and environmental strains. Furthermore, the flaA size of the clinical strain of A. veronii bv. sobria was mainly 909, 924, and 939 bp, and the size of A. hydrophila was 909 bp. The swimming motility of clinical isolates of both species was lower than the environmental isolates; however, the biofilm forming ability of the clinical isolates was high. Thus, the clinical isolates of A. veronii bv. sobria and A. hydrophila had different genetic and functional characteristics of flagellin than the environmental isolates. The characteristics of flagellin could serve as indicators to distinguish between clinical and environmental isolates of the both species. It may contribute to diagnosis of these diseases and the monitoring of clinical strain invasion into the natural environment.

Successful Endovascular Repair of a Kommerell's Diverticulum and a Right-Sided Aortic Arch.

BACKGROUND: A 57-year-old woman was diagnosed with Kommerell's diverticulum in the setting of a right aortic arch on computed tomography. CASE REPORT: Although asymptomatic, the maximum diameter of the aneurysm was 55 mm; thus, she underwent surgery to prevent rupture of the aneurysm. A bypass was constructed from the left common carotid artery to the left subclavian artery. A stent-graft was deployed from the distal right subclavian artery, and coil embolization of the diverticulum was performed via the left subclavian artery. She was discharged after 12 days of surgery. The postoperative four-month follow up showed a smaller aneurysm. CONCLUSION: Thoracic endovascular aortic repair is feasible and effective for Kommerell's diverticulum.

Retrograde Type A Aortic Dissection after Thoracic Endovascular Aortic Repair for Type B Dissection.

BACKGROUND: A 64-year-old woman presented with dilatation of the distal aortic arch secondary to chronic type B aortic dissection. CASE REPORT: The patient underwent fenestrated thoracic endovascular aortic repair (TEVAR) for closure of the entry site, and reconstruction of the left subclavian artery with a covered stent. On the 40th postoperative day, a retrograde type A aortic dissection (RTAD) was observed on computed tomography and she underwent emergency surgery. The entry tear, related to the proximal bare metal stent, was located in front of the aortic arch. A partial aortic arch replacement was performed. CONCLUSION: Consideration of the risk factors of RTAD is important when performing TEVAR.

A survey of extended-spectrum ß-lactamase-producing Enterobacteriaceae in environmental water in Okinawa Prefecture of Japan and relationship with indicator organisms.

Surveys of extended-spectrum ß-lactamase-producing Enterobacteriaceae (ESBL-pE) in stream water and untreated wastewater were carried out in Okinawa Prefecture, Japan. Thirty-six samples of water were collected from 18 streams in Okinawa Prefecture, as well as ten samples of wastewater flowing into four wastewater treatment plants (WWTPs). We investigated bacterial species, Escherichia coli O antigen, ESBL phenotype, ESBL genotype, and pulsed-field gel electrophoresis (PFGE) type of isolates, and total viable count and fecal coliforms as indicator organisms. The relation between indicator organisms and ESBL-pE was also validated using the same samples. A total of 141 ESBL-pE including 107 E. coli, 15 Klebsiella pneumoniae, 2 Proteus mirabilis, and 17 other species was isolated from stream water and wastewater. Of the 141 ESBL-pE, 14.9% and 54.6% were found to be blaCTX-M-15 and blaCTX-M-14-like types, respectively, which have been found in hospital isolates in Okinawa. Two pairs of possibly related patterns according to PFGE criteria were isolated from stream water and wastewater in two districts. When ESBL-pE was significantly isolated, total viable count and fecal coliform boundaries were ≥ 6.0 × 103 CFU/ml and ≥ 4.3 × 102 most probable number/100 ml, respectively. These results suggested that ESBL-pE isolated from stream water is human derived, and that total viable count and fecal coliforms will be useful as indicators for confirming the spread of ESBL-pE to the environment by means of simple hygiene surveys.

[A Case of Cecum Cancer with Synchronous Adrenal Solitary Metastasis].

We herein report a case of cecum cancer with synchronous adrenal solitary metastasis. A 62-year-old woman who had been treated for other disease complained of weight loss. Lower endoscopy revealed cecum cancer, and computed tomography( CT)demonstrated a solitary left adrenal tumor; thus, she was diagnosed with a metastatic tumor. We concluded that the patient was a possible candidate for surgical resection because she did not present with local metastasis other than in the adrenal glands. Ileocecal resection and left adrenalectomy were performed. The histological findings indicated moderately differentiated adenocarcinoma, which was compatible with cecum cancer. The patient was administered chemotherapy containing mFOLFOX6, and no recurrence has been detected 4 years after the surgery. Some patients who develop solitary adrenal metastasis from colonic cancer appear to be good candidates for surgery in anticipation of a good prognosis.

Sanitary evaluation of domestic water supply facilities with storage tanks and detection of Aeromonas, enteric and related bacteria in domestic water facilities in Okinawa Prefecture of Japan.

To provide for temporary restrictions of the public water supply system, storage tanks are commonly installed in the domestic water systems of houses and apartment buildings in Okinawa Prefecture of Japan. To learn more about the sanitary condition and management of these water supply facilities with storage tanks (hereafter called "storage tank water systems") and the extent of bacterial contamination of water from these facilities, we investigated their usage and the existence of Aeromonas, enteric and related bacteria. Verbal interviews concerning the use and management of the storage tank water systems were carried out in each randomly sampled household. A total of 54 water samples were collected for bacteriological and physicochemical examinations. Conventional methods were used for total viable count, fecal coliforms, identification of bacteria such as Aeromonas, Enterobacteriaceae and non-fermentative Gram-negative rods (NF-GNR), and measurement of residual chlorine. On Aeromonas species, tests for putative virulence factor and an identification using 16S rRNA and rpoB genes were also performed. Water from the water storage systems was reported to be consumed directly without boiling in 22 of the 54 houses (40.7%). 31 of the sampled houses had installed water storage tanks of more than 1 cubic meter (m3) per inhabitant, and in 21 of the sampled houses, the tank had never been cleaned. In all samples, the total viable count and fecal coliforms did not exceed quality levels prescribed by Japanese waterworks law. Although the quantity of bacteria detected was not high, 23 NF-GNR, 14 Enterobacteriaceae and 5 Aeromonas were isolated in 42.6%, 7.4% and 3.7% of samples respectively. One isolated A. hydrophila and four A. caviae possessed various putative virulence factors, especially A. hydrophila which had diverse putative pathogenic genes such as aer, hlyA, act, alt, ast, ser, and dam. Many bacteria were isolated when the concentration of residual chlorine was below 0.1 mg/l and the water temperature was above 20 °C. These results suggest that elevated water temperature and mismatch between tank size and water demand lead to loss of residual chlorine in tap water. Therefore, to minimize growth of aquatic bacteria such as Aeromonas spp. and Pseudomonas spp., we recommend that an appropriate size tank and/or volume of stored water is always used, and also suggest installation of some means of reducing water temperature such as shading.

Effects of Oral Antidiabetic Drugs on Changes in the Liver-to-Spleen Ratio on Computed Tomography and Inflammatory Biomarkers in Patients With Type 2 Diabetes and Nonalcoholic Fatty Liver Disease.

PURPOSE: Oral antidiabetic drugs (OADs) such as pioglitazone and metformin have beneficial effects in patients with nonalcoholic steatohepatitis. We prospectively assessed the effects of OADs on nonalcoholic fatty liver disease (NAFLD) in 886 men with type 2 diabetes mellitus and in a murine model of NAFLD. METHODS: Patients were randomized to receive pioglitazone, metformin, sitagliptin, or a non-OAD (control) for 6 months. All the patients received dietary and exercise guidance once a month during this study. Changes in the liver-to-spleen ratio on computed tomography (CT) and NAFLD-related parameters were measured from baseline to the end of treatment. FINDINGS: The liver/spleen ratio improved significantly in the pioglitazone and metformin groups compared with the control group (both P < 0.01), but not in the sitagliptin group (P = 0.73). The mean changes from baseline were -3.464 ± 10.156%, 19.236 ± 9.896%, 4.783 ± 1.467%, and 1.328 ± 0.802% in the control, pioglitazone, metformin, and sitagliptin groups, respectively. Multivariable analysis showed that the liver/spleen ratio was strongly correlated with high-sensitivity C-reactive protein concentration in the pioglitazone group (F = 9.973; P < 0.01) and abdominal visceral fat volume in the metformin group (F = 6.049; P < 0.05). CONCLUSIONS: Pioglitazone elicited the greatest improvements in features of NAFLD in type 2 diabetes mellitus. (Trial Registration: www.isrctn.org/, ISRCTN33414972, http://www.isrctn.org/).

Distribution of Aeromonas species in environmental water used in daily life in Okinawa Prefecture, Japan.

OBJECTIVES: The genus Aeromonas is known to causes diseases such as food poisoning, sepsis, and wound infection. However, the mode of Aeromonas transmission from environment to humans is not clearly understood. To evaluate the health risks of Aeromonas spp. in environmental freshwater, the number, proportion and putative virulence factors of Aeromonas species were investigated in Okinawa Prefecture, Japan. METHODS: Environmental freshwater samples were collected from three dams, two springs and three private wells. Aeromonas strains were identified by the biochemical method and the viable count was calculated. The production of extracellular enzymes and the virulence genes were investigated for possessing putative virulence factors using representative isolates. RESULTS: At least seven species of already-known Aeromonas isolates as well as unidentified Aeromonas spp. with/without arginin dehydrolase (ADH) exist in water at these sites. Aeromonas spp. was found to exist at over 1000 CFU/100 ml in one spring and two wells. A. veronii biovar sobria and A. jandaei were the predominant species in dams, and A. hydrophila and/or A. eucrenophila were predominant in wells. Almost all the sampled Aeromonas species produced protease, gelatinase, lipase, esterase and DNase, but A. caviae, A. caviae-like bacteria, and A. eucrenophila had low hemolytic activity. Most sampled A. hydrophila strains possessed both aerolysin gene (aer) and hemolysin gene (hlyA), but A. caviae and A. eucrenophila strains did not possess either gene. CONCLUSIONS: Since these results indicated that several Aeromonas species having potential pathogenicity exist in environmental water in Okinawa, surveys are recommended as a public health measure.

Current status of extended spectrum ß-lactamase-producing Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis in Okinawa prefecture, Japan.

Enterobacteriaceae producing extended spectrum ß-lactamase (ESBL) are distributed worldwide. In this study, 114 ESBL-producing Enterobacteriaceae were isolated by analyzing 1672 clinical isolates of Enterobacteriaceae collected from an Okinawa prefectural hospital in Japan between June 2013 and July 2014. The overall prevalence of ESBL-producing Enterobacteriaceae was 6.8%; the prevalence of different bacterial species among the ESBL-producing isolates was as follows: 11.5% Escherichia coli (90 of 783 isolates), 6.2% Klebsiella pneumoniae (19 of 307 isolates), and 11.1% Proteus mirabilis (5 of 45 isolates). The ESBL types blaCTX-M-1, -3, -15, -2, -14, -27, and mutants of blaSHV-1 were detected. Among them, blaCTX-M-15 (33.3%), blaCTX-M-14 (27.8%) and blaCTX-M-27 (33.3%) were dominant in the E. coli isolates, whereas a blaSHV mutant which possessed four mutations (Tyr7Phe, Leu35Gln, Gly238Ser and Glu240Lys) in the amino acid sequence of SHV-1 dominated in the K. pneumoniae isolates (11 of 19, 57.9%). The pandemic E. coli ST131 clone was found to constitute 3.3% of the overall examined isolates and 62.2% of the ESBL-producing E. coli isolates. Our results suggest that the genetic combination of blaCTX-M, and blaSHV and antibiotics-resistant profile were different from that in other regions such as other areas of Japan, Asia, Europe, and North America, especially in the ESBL-producing K. pneumoniae isolates and in the E. coli B2-O25b-ST131 isolates possessing blaCTX-M-15 (40.7% of the E. coli B2-O25b-ST131 isolates). Taken together, our results indicate that the ESBL-producing Enterobacteriaceae in Okinawa, Japan, might be of a unique nature.

Use of a modified early warning score system to reduce the rate of in-hospital cardiac arrest.

BACKGROUND: Physiological abnormalities are often observed in patients prior to cardiac arrest. A modified early warning score (MEWS) system was introduced, which aims to detect early abnormalities by grading vital signs, and the present study investigated its usefulness. METHODS: Based on previous reports, the Chubu Tokushukai Hospital-customized MEWS was developed in Okinawa, Japan. The MEWS was calculated among all inpatients, and the rates of in-hospital cardiac arrests (IHCAs) were compared according to the score. The warning zone (WZ) was set as 7 or more because of the high possibility of acute deterioration. The MEWS system was introduced to provide immediate interventions for patients who reached the WZ in accordance with the callout algorithm. The numbers of IHCAs were compared between the 18 months before and after introduction of the MEWS system. RESULTS: The numbers of patients who experienced IHCA with each score were as follows: score of 6, 1 of 556 patients (0.18 %); score of 7, 4 of 289 (1.40 %); score of 8, 2 of 114 (1.75 %); and score of 9 or more, 2 of 56 (3.57 %). There was no significant difference in the mean age or sex between before and after the introduction of the MEWS system. The rate of IHCAs per 1000 admissions decreased significantly from 5.21 (79/15,170) to 2.05 (43/17,961) (p < 0.01). CONCLUSIONS: The Chubu Tokushukai Hospital-customized MEWS was applied to all inpatients, and the rate of IHCA decreased owing to the introduction of the system, as the system enables early interventions for patients who have the possibility of acute deterioration.

Nonsurgical treatment of scalp arteriovenous malformation using a combination of ultrasound-guided thrombin injection and transarterial coil embolization.

Scalp arteriovenous malformations are treated by surgical excision in many patients. We report a patient with a scalp arteriovenous malformation who was successfully treated by a combination of ultrasound-guided thrombin injection (UGTI) and transarterial coil embolization. This patient was a 52-year-old man with a subcutaneous mass in the left retroauricular region. An angiogram showed that the mass was a nidus of arteriovenous malformation. We performed UGTI after transarterial coil embolization. No recurrence or complication was reported during 2 years of follow-up. This report describes the advantages of UGTI and the method for complete occlusion of the collateral artery.

Cardioprotective effects of tetrahydrobiopterin in cold heart preservation after cardiac arrest.

BACKGROUND: It has recently been shown that tetrahydrobiopterin (BH4), an essential cofactor of nitric oxide synthase (NOS), reduces ischemia-reperfusion myocardial injury. The aim of this study was to determine if supplementation with BH4 after cardiac arrest followed by cold heart preservation would exert a cardioprotective effect against ischemia-reperfusion injury. MATERIALS AND METHODS: Isolated perfused rat hearts were subjected to 4 degrees C cold ischemia and reperfusion. Hearts were treated with cold cardioplegic solution with or without BH4 just before ischemia and during the first 5 min of reperfusion period. Effects of BH4 on left ventricular function, myocardial contents of high-energy phosphates, and nitrite plus nitrate were measured in the perfusate, before ischemia and after reperfusion. Moreover, the effect of BH4 on the cold-heart preservation followed by normothermic (37 degrees C) ischemia was determined. RESULTS: BH4 improved the contractile and metabolic abnormalities in reperfused cold preserved hearts that were subjected to normothermic ischemia. Furthermore, BH4 significantly alleviated ischemic contracture during ischemia, and restored the diminished perfusate levels of nitrite plus nitrate after reperfusion. CONCLUSION: These results demonstrated that BH4 reduces ischemia-reperfusion injury in cold heart preservation. The cardioprotective effect of BH4 implies that BH4 could be a novel and effective therapeutic option in the preservation treatment of donor heart after cardiac arrest.

Graft for mycotic thoracic aortic aneurysm: omental wrapping to prevent infection.

Nine cases of mycotic thoracic aortic aneurysm were treated surgically between July 1995 and March 2003. The aneurysms were located in the ascending aorta in 1 patient, the descending thoracic aorta in 5, and the thoracoabdominal aorta in 3. Preoperatively, 3 patients were in shock due to rupture of the aneurysm. All patients underwent aneurysmectomy and in-situ graft placement. In 5 patients, the graft was covered with a pedicled omental flap to prevent postoperative graft infection. There were 2 hospital deaths: one patient died of multi-organ failure, and the other died from intrathoracic bleeding. After discharge, one patient died from intrathoracic bleeding 3 months after surgery. These 3 patients had not received omental wrapping. Postoperative graft infection did not occur in the 6 surviving patients during a mean follow-up period of 4.0 +/- 3.1 years. It was concluded that covering the prosthetic graft with a pedicled omental flap may help prevent postoperative graft infection and improve the surgical results.

Improvement of liver function after surgery for Budd-Chiari syndrome.

PURPOSE: We evaluated the relationship between liver histology and postoperative improvement of liver function after surgery for Budd-Chiari syndrome (BCS). METHODS: Over a period of 23 years, we operated on 46 patients with BCS by reconstructing the occluded inferior vena cava (IVC) and reopening as many occluded hepatic veins as possible. We divided the patients into a liver cirrhosis group (group I, n = 30) and a hepatic fibrosis or liver congestion group (group II, n = 16), and compared the ages, duration of illness, preoperative liver function, changes in liver function, and changes in esophageal varices (EV). RESULTS: There were no hospital deaths. In group I the patients were older, and the duration of illness was longer. The group I patients also had a lower thrombotest percentage and a higher serum ammonia. The indocyanine green clearance (ICG) test showed more remarkable improvement in liver function in group II. The rate of disappearance of EV was also higher in group II. CONCLUSION: Surgery during the early stage of BCS is important in improving postoperative liver function.

Malignant fibrous histiocytoma of the heart complicated by factor XI deficiency in a Jehovah's Witness patient.

We describe a 16-year-old girl with malignant fibrous histiocytoma (MFH) of the heart complicated by factor XI deficiency. The preoperative diagnosis was left atrial myxoma. We decided to perform the operation owing to a normal bleeding time. Operative findings suggested a malignant tumor. The patient was a Jehovah's Witness, and extensive excision was not performed because blood transfusion was not allowed. We resected as much of the tumor and left atrial appendage as possible. The pathologic diagnosis was MFH. Excessive bleeding was not observed during the operation. Bleeding time helps to determine whether a surgical procedure is indicated in patients with factor XI deficiency.

The prevention of nerve injury in aortic arch aneurysmal surgery.

In a case of aortic arch aneurysm associated with adhesion to the surrounding structures, we devised an operative technique to avoid nerve injury during the surgical procedure. By preserving the adventitial layer of the aortic arch aneurysm to which the phrenic and recurrent nerves were attached, injury to the nerves was avoided, and the aneurysmectomy was completed with the distal anastomosis being performed intraluminally.

Diaphragmatic plication in adult patients with diaphragm paralysis after cardiac surgery.

OBJECTIVE: We investigated the benefit of diaphragmatic plication for weaning from mechanical ventilation in these adult patients. PATIENTS AND METHODS: Four patients underwent diaphragmatic plication for difficulty of weaning from mechanical ventilation due to diaphragmatic paralysis. They were all men with an average age of 70.5 +/- 6.3 years. Three of the patients had undergone cardiac surgeries for coronary artery bypass grafting and one patient ascending aortic replacement for pseudoaneurysm after coronary revascularization. Right diaphragmatic plication (muscle sparing procedure) was performed between 30 to 61 days after cardiac surgery. RESULTS: The mean forced tidal volume improved dramatically from 216 to 415 ml after plication in all patients, and it was possible to discontinue mechanical ventilation from 2 to 12 days after plication. One patient with obstructive respiratory dysfunction died from aspiration pneumonia 15 days after plication. However, postoperative tidal volume in this patient improved to 420 ml and he was able to be weaned from ventilatory support five days after plication. The other three patients were discharged between 26 to 58 days after plication and continue to do well without symptoms. CONCLUSION: Diaphragmatic plication is a useful procedure for treatment of diaphragmatic paralysis in adults as well in children.

Direct cannulation of the common carotid artery during the ascending aortic or aortic arch replacement.

OBJECTIVE: To evaluate cerebral perfusion using direct cannulation into the common carotid artery. A new technique is needed to protect brain ischemic injury during ascending aortic or aortic arch replacement. METHODS: This technique was evaluated for patients who would have difficulty maintaining adequate cerebral perfusion during surgery. The procedure was performed when patients had the following diagnoses: pseudoaneurysm formation in contact with the sternum with the risk of aneurysmal rupture (n = 5), acute aortic dissection with compression of the true lumen of the innominate artery by the pseudolumen (n = 3), or a large volume of thrombus in the lumen of the aneurysm with the risk of cerebral thromboembolism if standard extracorporeal circulation was used (n = 2). The perfusion catheter was cannulated into one side of the common carotid artery (right side: n = 6, left side: n = 4) and mean perfusion flow rate was found to be 175 mL/min. The operative procedures consisted of ascending aortic and aortic arch replacement with coronary artery bypass grafting in six patients, ascending aortic replacement in 2 patients, and innominate artery reconstruction/innominate artery and right subclavian artery reconstruction in one patient. RESULTS: No cerebral accidents or deaths occurred while patients were hospitalized. We have followed up patients for a mean of 2.1 years (maximum 3.6 years), with no complications noted from the surgical procedure. CONCLUSIONS: Direct cannulation of the common carotid artery is a simple, safe, and acceptable cerebral protection for patients undergoing aortic or aortic arch replacement procedures in the patients with these specific conditions.

Type B dissection complicated with subacute visceral ischemia.

A 63-year old man presented with back pain and abdominal pain that worsened after eating. Contrast-enhanced computed tomography revealed type B aortic dissection. Arteriography 20 days after dissection revealed celiac trunk stenosis and the superior mesenteric artery did not arise from the true lumen. Saphenous vein bypass grafting from the right common iliac artery to the superior mesenteric and gastroduodenal arteries was performed. The postoperative course was uneventful and the abdominal symptoms completely disappeared.