Diversity and characterization of oxytetracycline-resistant bacteria associated with non-native species, white-leg shrimp (Litopenaeus vannamei), and native species, black tiger shrimp (Penaeus monodon), intensively cultured in Thailand.

AIMS: This study aimed at surveying prevalence of oxytetracycline (OTC)-resistant bacteria in the white-leg shrimp Litopenaeus vannamei, and the black tiger shrimp Penaeus monodon, intensively cultured in Thailand. We investigated the phylogenetic diversity of the bacterial isolates, as well as the minimum inhibitory concentration (MIC) of OTC, the occurrence of major OTC-resistant genes and multiple-antibiotic resistance in the isolates. METHODS AND RESULTS: Shrimps were collected from culture ponds, and the homogenates of whole bodies were plated on tryptic soy agar supplemented with or without OTC. Percentages of OTC-resistant bacteria were 0·3-52·1% in white-leg samples and 0·008-22·3% in black tiger samples. Analyses of 16S rDNA sequences indicated that most OTC-resistant isolates were closely related to Aeromonas spp. and Lactococcus garvieae. MICs of OTC were 4-128 µg ml(-1) in the OTC-resistant aeromonads and 128-256 µg ml(-1) in OTC-resistant L. garvieae. OTC resistance was found to be conferred by the genes tet(A), tet(C), tet(D), tet(E), tet(M) and tet(S), detected either singly or in pairs. No resistance to ceftazidime, imipenem or chloramphenicol was observed in any isolate. CONCLUSIONS: Both species of shrimp are associated with OTC-resistant bacteria, occasionally at high densities exceeding 10(6) cfu g(-1). The associated bacteria, predominantly Lactococcus and Aeromonas genera, are potential pathogens and are reservoirs of a variety of OTC-resistant genes. SIGNIFICANCE AND IMPACT OF THE STUDY: Cultured shrimps can be vehicle to carry OTC-resistant bacteria to domestic and foreign consumers via the food chain. Very low populations of OTC-resistant bacteria observed in the several ponds suggest that levels of the resistant bacteria are artificially high and should be reduced in farmed shrimps.

Role of inhaled tiotropium on the perioperative outcomes of patients with lung cancer and chronic obstructive pulmonary disease.

BACKGROUND: Tiotropium, a long-acting bronchodilator, can be used perioperatively in patients with lung cancer and airway obstruction, although its benefits for the perioperative outcome remain unclear. METHODS: We prospectively collected the perioperative data of 44 patients with resectable lung cancer and untreated airway obstruction. Tiotropium was not used before September 2007 (control group, n = 24) but was used routinely thereafter (treated group, n = 20). We estimated a propensity score to adjust comparisons between the groups. RESULTS: Tiotropium improved preoperative global pulmonary function significantly, especially in four patients. Postoperative outcomes in these major responders were significantly better than those in the remaining minor responders. However, postoperative outcomes were not significantly different between the treated group (n = 15) and the control group (n = 15) matched by a propensity score. CONCLUSION: Regardless of its favorable effects on preoperative pulmonary function, we could not establish a significant benefit of tiotropium for postoperative outcomes overall. Nonetheless, our data suggested that tiotropium might have improved the postoperative outcomes of major responders.

[Our method of exposing the femoral artery and vein and the axillary artery].

A thoracic surgeon must have the skill to expose the femoral artery and vein, as well as the axillary artery, quickly without causing any complications. We briefly describe our method of exposing these vessels. We devised this method to reach the targeted blood vessel within the shortest time and distance, based on a sound knowledge of the surgical anatomy. To prevent postoperative lymphorrhoea. we ligate the lymphoducts during the operation.

Effect of glutathione on the development of rat embryos following microinsemination.

PURPOSE: The present study was carried out to evaluate the role of glutathione on rat embryo developmental potential after ICSI. We observed the effects of glutathione on the development of non-treated rat embryos, ICSI embryos and embryos with sham injection treatment. The development of glutathione-microinjected embryos was also observed. METHODS: Oocytes and fertilized embryos were obtained from superovulated Wistar-Imamichi rats and cultured in mR1ECM medium. Oocytes and embryos were then allowed to develop to assess the effect of glutathione on the development rate in intact embryos, micro-injected embryos and ICSI embryos. RESULTS: (1) In the intact embryo, the proportion of blastocyst stage development increased when 0.01 mM GSH was added to the medium compared to the control. (2) Microinjection of glutathione (GSSG, GSH) into the embryo increased development at each stage, and the addition of 0.2 nM GSSG or GSH significantly increased blastocyst development, in comparison to that of the control (P < 0.05). (3) Compared to the control, all the GSSG and GSH concentrations improved damaged blastocyst development, where 0.01 mM GSH improved significantly (P < 0.05). (4) The addition of glutathione in the medium increased the rate of blastocyst development after ICSI. A significantly higher number of TE and total cells were obtained in the micro-injected embryo with both of the 0.02 mM GSSG and GSH treatments (P < 0.05). CONCLUSIONS: The addition of glutathione into the culture media can improve early embryo development and is capable of repairing the damage of ICSI rat embryos.

The beneficial effect of fructose and glucose on in vitro maturation and the fertilization of porcine oocytes.

PURPOSE: This study was conducted to investigate the effects of supplementing fructose in the culture medium on in vitro maturation (IVM), in vitro fertilization (IVF), and metabolism of porcine oocytes. METHODS: Porcine oocytes were matured in vitro in modified North Carolina State University 37 medium (NCSU-37) and then supplemented with either glucose (5.5 mM), fructose (5.5 mM), or glucose (2.75 mM) plus fructose (2.75 mM). The maturation and fertilization of oocytes, and the incorporation and oxidation of 14C-glucose, 14C-fructose, and 14C-methionine in oocytes at different stages of development were examined. RESULTS: The supplementation of glucose plus fructose significantly promoted (P < 0.05) oocytes germinal vesicle break down (GVBD), maturation to metaphase II (MII), penetration by spermatozoa, and male pronuclear formation compared with glucose. The incorporation and oxidation of 14C-methionine into the oocyte significantly increased (P < 0.05) with glucose plus fructose supplementation than glucose. A significantly higher (P < 0.05) rate of incorporation and oxidation was achieved with 14C-fructose compared to 14C-glucose. CONCLUSIONS: Glucose plus fructose supplementation improved maturation, penetration by spermatozoa, male pronuclear formation, and energy metabolism by porcine oocytes.

[Mitral valve replacement with St. Jude Medical mechanical heart valve in severely calcified mitral annulus; a simple and novel technique].

A 63-year-old woman who had been on hemodialysis for 20 years was referred to our institution to undergo mitral valve surgery for mitral valve stenosis. Intraoperative investigations confirmed severe calcification of the mitral leaflets and mitral annulus. We excised the only anterior leaflet, but preserved the posterior leaflet to prevent a fatal complication such as posterior left ventricular rupture or injury of the coronary artery. We passed 2-0 polyester mattress sutures through the anterior mitral annulus from the left ventricle to the left atrium, and then folded the posterior leaflet. Preserved posterior leaflet was fixed to posterior mitral annulus and prosthetic valve. The mitral valve was replaced using a St. Jude Medical mechanical heart valve with a specific structure, whose hinge shifts to the left atrial side and most of the leaflet to move its housing. This particular structure enables to perform this procedure without the need for excision of a severely calcified posterior mitral leaflet and annulus.

Prebiotic effect of two grams of lactulose in healthy Japanese women: a randomised, double-blind, placebo-controlled crossover trial.

Sixty healthy Japanese women with a defaecation frequency of 2-4 times/week participated in this randomised, double-blind crossover trial. Participants received 2 g/day lactulose for 2 weeks and placebo in a random order, separated by a washout period of 3 weeks. Eight participants were excluded who did not satisfy the conditions, and therefore data from 52 were analysed. The primary outcome was defaecation frequency and the secondary outcomes were the number of defaecation days, faecal consistency, faecal volume, and the number and percentage of Bifidobacterium in faeces. The defaecation frequency (times/week) was significantly higher during lactulose (4.28±0.23) than placebo (3.83±0.23) treatment (delta (Δ) 0.45 [95% confidence interval (CI) 0.10-0.80], P=0.013). The defaecation days (days/week) was significantly higher during lactulose (3.77±0.17) than placebo (3.47±0.17) treatment (Δ0.30 [95% CI 0.04-0.56], P=0.024). Faecal consistency using the Bristol Stool Scale (/defaecation) was significantly higher during lactulose (3.84±0.10) than placebo (3.68±0.10) treatment (Δ0.16 [95% CI 0.00-0.31], P=0.044). Faecal volume (/week) was significantly higher during lactulose (21.73±3.07) than placebo (17.65±3.07) treatment (Δ4.08 [95% CI 0.57-7.60], P=0.024). The number of Bifidobacterium in faeces (log colony forming units/g faeces) was significantly higher during lactulose (9.53±0.06) than placebo (9.16±0.06) treatment (Δ0.37 [95% CI 0.23-0.49], P<0.0001). The percentage of Bifidobacterium in faeces was also significantly higher during lactulose (25.3±1.4) than placebo (18.2±1.4) treatment (Δ7.1 [95% CI 2.9-11.4], P=0.0014). Finally, straining at defaecation (/defaecation) during lactulose (3.62±0.24) treatment was significantly lower than during placebo (3.97±0.24) treatment (Δ0.35 [95% CI -0.69 - -0.02], P=0.037). No significant difference was observed between lactulose and placebo with regard to flatulence. Severe adverse effects did not occur. Thus, oral ingestion of 2 g/day lactulose had a prebiotic effect, increasing the number and percentage of bifidobacteria in faeces, softening the faeces, and increasing defaecation frequency, but without increasing flatulence.

Genomic organization, expression and evolution of porcine CRSP1, 2, and 3.

Recently we identified and characterized porcine calcitonin receptor-stimulating peptide (CRSP) 1, CRSP2 and CRSP3 as members of the calcitonin/calcitonin gene-related peptide (CT/CGRP) family. In the present study, the genomic sequences and organization of CRSP1, 2, and 3 were determined, and the expression of the genes in the porcine brain was investigated using in situ hybridization. Analysis of 5'-upstream regions of the three CRSPs demonstrated that CRSP1 and CRSP2 have almost identical sequences (>98% similarity) and high sequence similarities including functional transcription binding sites with the corresponding region of human CALCA (CT/alpha CGRP), whereas CRSP3 retains less similarity with the above genes. RH mapping of CRSPs demonstrated that they resided in a region of swine chromosome 2 (SSC2). The arrangement of the genes in the region was found to be conserved in corresponding human and mouse regions. In situ hybridization demonstrated sense transcripts of the three genes in cerebrum, hippocampus, hypothalamus, pons/midbrain, and thalamus of 3-month-old pigs, and CRSP2 sense transcripts additionally in tractus opticus. The sense transcripts of alpha CGRP and CALCB (beta CGRP) were detected in cerebrum, hippocampus, and pons/midbrain of newborn mice, and to a lesser extent in pons/midbrain of 8-week-old mice. These results taken together with the chromosomal conservation and phylogenetic clustering of CT/CGRP family indicate that CRSP1, 2, and 3 may be functionally different from alpha CGRP and beta CGRP, though they are indicated to have a common progenitor gene.

[Subdural hematoma following cardiac surgery].

We report 3 cases of subdural hematoma following open heart surgery under cardiopulmonary bypass. In 2 patients, emergency removal and drainage of a subdural hematoma was performed by neurosurgeons, and conservative management was performed in the remaining one. All patients belonged to a high risk group of bleeding due to anticoagulation therapy, and they had no episode of head trauma. Subdural hematomas may have been due to rapid alterations in cerebral volume, leading to a tearing of the dural bridging veins under cardiopulmonary bypass. Although early diagnosis and prompt treatment is very important, it is often difficult to examine neurological findings after cardiac surgery. We consider that patients who need long-term sedation under anticoagulation therapy must have their neurological status checked at least once in a few days.

Peculiar clinical features of cellulitis in peripheral lymphedema.

Although the occurrence of cellulitis in lymphedema (LE) is believed to be an infection-related event, many findings in its clinical course seem to suggest that it is unlikely to be an infection. Therefore, we tried to clarify the specific features of cellulitis in LE. In-hospital courses of cellulitis obtained from medical charts were reviewed in the patients with leg LE (LE; 24 patients, 72admissions), chronic venous insufficiency (CVI; 28 patients, 29 admissions), and leg cellulitis secondary to wound infection without underlying disease (N; 42 patients, 42 admissions). The patients with LE complained of less local pain (peak numerical scale; LE: 1.4 ± 1.7, CVI: 4.1 ± 2.5, N: 3.2 ±2.0, p < 0.0001), showed an abnormally higher peak procalcitonin level (LE: 33.8 ± 34.8 (N = 7), CVI: 2.9 ± 5.8 (N = 8), N: 0.4 ± 0.6(N = 10), p < 0.05), and required fewer antibiotics (LE: 1.1 ± 0.3, CVI: 1.8 ± 0.9, N: 1.5 ± 0.9, p < 0.0001). These findings suggested that the occurrence of cellulitis in LE seems unlikely to be an infection-related type of cellulitis similar to that found in CVI.

[Surgical repair of coarctation of the aorta in adults].

We report the surgical treatment of coarctation of the aorta (CoA) in 3 adults, 2 women and 1 man, aged between 18 and 32 years old. All of the patients had blood pressure gradients higher than 70 mmHg between the upper and lower limbs. In 2 patients, we simply clamped the aorta and excised the CoA: while in the other patient, we excised the CoA using partial extracorporeal circulation with a femoro-femoral (F-F) bypass. Reconstruction was done by an end to end anastomosis in 2 patients and with an artificial tube graft in 1 patient who regulred the extended aortic arch repair. Postoperatively, the pressure gradients between the upper and lower limbs dropped to below 20 mmHg in intensive care unit (ICU). Two of the patients have now stopped taking antihypertensive drugs and the other patient is taking half the preoperative dose.

[The Waffle procedure for constrictive epicarditis after direct closure of atrial septal defect; report of a case].

A 20-year-old man was admitted with a diagnosis of constrictive pericarditis 6 months after direct closure of atrial septal defect (ASD). He complained of fatigue and dyspnea. Cardiac echo cardiography, computed tomography (CT), magnetic resonance imaging (MRI) and cardiac catheterization suggested pericardial and epicardial constriction. During the operation, the thickened pericardium was peeled off. Multiple longitudinal and transverse incisions were made in the thickened epicardium as reported by waffle. Postoperative hemodynamic state was improved. The cardiac index increased from 1.91 to 3.17 l/min/m2. The pulmonary capillary wedge pressure (PCWP) decreased from 26 to 14 mmHg, although dip and plateau pattern was maintained. The postoperative course was uneventful.

[Aortic root abscess after percutaneous coronary intervention].

A 68-year-old man, who had undergone percutaneous coronary intervention for right coronary disease 2 weeks earlier, was admitted to our hospital for investigation of a fever. Blood culture and echocardiography revealed isolated aortic valve infective endocarditis. He was treated with antibiotics for more than 1 week, but echocardiography showed an aortic root abscess with severe aortic regurgitation. Thus, we performed aortic root replacement using an artificial Freestyle stentless bioprosthesis valve. The patient had an uneventful postoperative course and antibiotic treatment was continued for a further 8 weeks.

Galacto-oligosaccharides ameliorate dysbiotic Bifidobacteriaceae decline in Japanese patients with type 2 diabetes.

Gut microbiota affects the host's metabolism, and it is suggested that there are differences in gut microbiota composition between patients with type 2 diabetes and healthy individuals. Additionally, dysbiosis may increase the concentration of lipopolysaccharides (LPS), causing metabolic endotoxemia, which induces impaired glucose tolerance. Several studies have reported relationships between metabolic diseases and the gut microbiota; and prebiotics, such as oligosaccharides, are commonly consumed to regulate gut microbiotas in healthy individuals. Galacto-oligosaccharides (GOS) are a major prebiotic, which specifically increase Bifidobacteriaceae abundance. Recent studies have reported that Bifidobacteriaceae improved metabolic endotoxemia or impaired glucose tolerance. However, there are few studies reporting the effects of GOS on patients with type 2 diabetes. In the current study, we compared clinical parameters, faecal gut microbiota, their associated metabolic products and their components such as LPS, and LPS-binding protein (LBP) produced by the host, between patients with diabetes and healthy controls. We then assessed the effects of GOS on glycaemic control, and gut microbiotas and metabolites in patients with type 2 diabetes in a double-blind controlled manner. LBP levels were significantly higher in patients with diabetes than those of healthy subjects, which was consistent with previous reports. The abundance of Bifidobacteriaceae and the diversity of intestinal microbiota were significantly lower in patients with diabetes than in healthy subjects. Interestingly, Bifidobacteriaceae was markedly restored in patients with diabetes after consumption of GOS, whereas LBP and glucose tolerance did not improve during this short-term trial period. In the present study, we demonstrated that GOS can ameliorate dysbiosis in patients with diabetes, and continuous intake of GOS may be a promising method for managing type 2 diabetes.

[Treatment for perioperative arrhythmias with nifekalant hydrochloride].

Ventricular tachycardia (VT), ventricular fibrillation (VF), and atrial flutter (AFL) are potentially fatal or serious complications arising after cardiac surgery. Generally, we treat these complications with class I antiarrhythmic agents and/or direct counter shock (DC). However, sometimes these complications do not respond to antiarrhythmic agents and require frequent DC. Moreover, these class I agents induce heart failure due to their negative inotropic effect. Nifekalant hydrochloride (NIF) is a class III antiarrhythmic agent that prolongs the refractory period of the atrial and ventricular myocardium without any negative inotropic action. From July 2003 to September 2004, we treated 11 patients with NIF for perioperative arrhythmias (VT 5, VF 2, and AFL 4). NIF was administered by continuous intravenous infusion (0.3 to 0.4 mg/ kg/h) to prevent the recurrence of VT/VF and AFL. NIF prevented the recurrence of VT in 3 of the 5 cases. No recurrence was observed in 2 cases with VF. Furthermore, NIF prevented the recurrence of AFL in all the 4 patients. None of the patients exhibited changes in heart rate, cardiac output, and QTc interval. Additionally, no occurrence of Torsades de pointes was observed in any of the cases. In conclusion, NIF is an effective and safe antiarrhythmic agent for the treatment of perioperative arrhythmias under continuous monitoring of the QTc interval.

[Mid-term results of overlapping cardiac volume reduction for ischemic cardiomyopathy].

BACKGROUND: Left ventricular restoration (LVR) is thought to be effective for ischemic cardiomyopathy (ICM). We assessed the early and mid-term results of overlapping cardiac volume reduction (OLCVR) as an operation for LVR on patients with ICM. METHODS: From February to June 2004, 4 patients with ICM underwent OLCVR in our department. RESULTS: There was no death. Significant decreases in the left ventricular end diastolic volume index and in the left ventricular end systolic volume index were observed in the early postoperative period in all the 4 patients. These effects were still evident 1 year later, with improved New York Heart Association (NYHA) functional class and a smaller left ventricular diastolic diameter. One patient required implantation of a cardioverter defibrillator 16 months postoperatively. CONCLUSION: The favorable early and mid-term results of the OLCVR indicate that it is an effective procedure for patients with ICM.

[Blunt right atrial rupture].

Atrial rupture without other major injury following blunt chest trauma is rare, but carries high mortality rate. Here we report 3 cases of blunt atrial rupture. All patients presented with hypotension and loss of consciousness. Echocardiography and computed tomography (CT) demonstrated pericardial effusion in all cases. All patients underwent surgery with a median sternotomy. Tear of the right atrium was seen in 2 patients, and no remarkable tear was seen in 1 patient. One of the 3 patients underwent surgery using cardiopulmonary bypass because he showed cardiogenic shock. Pericardiocentesis before the operation was not effective. All patients were discharged uneventfully. Echocardiography and CT were useful for diagnosis of cardiac rupture. In most reported cases, the cardiac tear has been repaired without using cardiopulmonary bypass. However, it is difficult to diagnose location of the tear, therefore, the repair became safer using cardiopulmonary bypass for the patients with cardiogenic shock. This article has described the mechanisms and generation of blunt chest trauma lesions.

Effect of three-size too large strong stocking on venous hemodynamics in normal subjects.

OBJECTIVES: To study the differences in impact on venous hemodynamics between larger size strong graduated elastic compression stockings (GECS) and appropriate size strong/moderate GECS. METHOD: In healthy legs fitted for a small (Group S; n = 8) and large (Group L; n = 8) GECS, air plethysmography was performed without GECS, with an appropriate size strong GECS (GECS1), with a three-size too large strong GECS (GECS2), and with an appropriate size moderate GECS (GECS3) in this order. RESULTS: In Group S, interface pressure with GECS2 was equal to or higher than that with GECS3. Decreased venous volume, unchanged ejection volume, and decreased residual volume were achieved by GECS, but differences in these parameters among GECS were not observed. Although insignificant, a similar tendency was found in Group L. CONCLUSIONS: A larger size strong GECS seemed to have equivalent interface pressure and impact on venous hemodynamics compared to an appropriate size moderate or strong GECS.

Up-regulation of inositol 1,4,5 trisphosphate receptor expression in atrial tissue in patients with chronic atrial fibrillation.

OBJECTIVES: We examined whether patients with atrial fibrillation (AF) have alterations in atrial inositol 1,4,5 trisphosphate receptors (IP3 receptors). BACKGROUND: Abnormal intracellular Ca2+ homeostasis occurs in chronic AF. The intracellular Ca2+ concentration is regulated by ryanodine and IP3 receptors. We recently reported alterations in ryanodine receptors in atrial tissue from patients in chronic AF. METHODS: We analyzed IP3 receptor expression in the right atrial myocardium from 13 patients with mitral valvular disease (MVD) with AF (MVD/AF), five patients with MVD who had normal sinus rhythm (MVD/NSR) and eight control patients with NSR (tissue obtained during coronary artery bypass surgery). Hemodynamic and echocardiographic data were obtained preoperatively, and an immunohistochemical study was performed on atrial tissue. RESULTS: The relative expression level of IP3 receptor protein was significantly greater in MVD/AF (0.75 +/- 0.26) than it was in MVD/NSR (0.42 +/- 0.13, p < 0.01), and both were significantly above control (0.14 +/- 0.08). The relative expression level of IP3 receptor messenger RNA was significantly greater in the MVD/AF group (0.028 +/- 0.008) than it was in the control group (0.015 +/- 0.004, p < 0.01), but patients with MVD/AF did not differ from patients with MVD/NSR (0.020 +/- 0.006). The relative expression levels of IP3 receptor protein and messenger RNA were higher in patients with left atrial dimension > or = 40 mm, pulmonary capillary wedge pressure > or = 10 mm Hg and right atrial pressure > or = 5 mm Hg. Inositol 1,4,5 trisphosphate receptors were over-expressed in the cytosol and at the nuclear envelope of atrial myocytes in MVD. CONCLUSIONS: Since chronic mechanical overload of the atrial myocardium increased IP3 receptor expression, especially in patients with chronic AF, up-regulation of IP3 receptors may be important in modulating intracellular Ca2+ homeostasis and initiating or perpetuating AF.

cDNA structure of an insulin-related peptide in the Pacific oyster and seasonal changes in the gene expression.

Insulin-related peptide cDNA was characterized in the Pacific oyster Crassostrea gigas. It was determined that three transcripts with differing lengths of 3'-untranslated region (3'-UTR) were expressed in the visceral ganglia. The insulin-related peptide cDNA contained a number of AUUUA motifs that were typical of adenylate/uridylate-rich elements in the 3'-UTR. The deduced preprohormone was a polypeptide of 161 residues and showed a conformation typical of preprohormones of the insulin superfamily, which included conserved amino acids necessary to adopt the globular insulin structure. The expression of the three different transcripts was variable throughout the year, with the highest expression observed in March and lower expression in November and July. The expression of the shortest mRNA in March was about tenfold higher than in July, while the expression of the longest transcript varied approximately twofold during the year. The accumulation of glycogen in the soft body rapidly increased in October and November, and robust body growth and gametogenetic development occurred in March to May. The period of the highest expression of the oyster insulin-related peptide gene corresponded to the onset of body growth and gametogenetic development, but did not overlap with the period of glycogen accumulation. This is the first report that fully details the structure and expression of the insulin-related peptide gene in bivalves.