Development of a high-performance liquid chromatographic glycated albumin assay using finger-prick blood samples.

BACKGROUND: Home blood glucose monitoring can be effective for the self-management of diabetic patients. Hemoglobin A1c (HbA1c) is a widely used marker that reflects the average blood glucose within 1-2 months but does not sensitively respond to behavioral changes. Self-monitoring of blood glucose, continuous glucose monitoring, and flush glucose monitoring are sensitive; however, the cost and invasiveness of these tests prevent their widespread use. We focused on glycated albumin (GA), which reflects the average blood glucose levels over 1-2 weeks, and established a GA measurement method for self-sampling, finger-prick blood, which may be submitted for testing through postal service to receive weekly results. METHODS: A high-performance liquid chromatography assay was established to measure GA levels in finger-prick blood samples from 103 diabetic patients and the results were compared with venous blood measurements using an enzymatic method. Furthermore, conditions for sending blood samples by mail were evaluated. Under these conditions, samples from 27 healthy and 32 patient volunteers sent through postal service were compared with samples stored in the laboratory. RESULTS: GA levels were measured in samples containing > 20 µg albumin, which resulted in a CV less than 0.3%. The correlation between the GA levels of finger-prick blood measured using HPLC and the GA levels of venous blood measured using the enzymatic method was R2 = 0.988 with the slope âˆ¼ 1.0, suggesting that the two were nearly equivalent. GA levels were stable for four days at 30 °C and two days at 37 °C. Mail-delivered samples exhibited a high correlation with samples that were not sent (R2 > 0.99). CONCLUSIONS: We established a method to measure GA levels in self-sampled, finger-prick blood sent through postal service in Japan. The method is applicable for weekly feedback of GA levels, which is potentially useful for motivating behavioral changes. In addition to markers such as HbA1c and blood glucose, GA can be used as a marker for assessing dietary and physical activities. This study highlighted the importance of GA monitoring by developing a suitable measurement method for weekly monitoring of GA levels.

The effects of continuous exposure to low-dose chlorine dioxide gas on the characteristics of induced pluripotent stem cells.

Background: Recently, various regenerative therapies have been developed based on induced pluripotent stem (iPS) cells. However, hygienic control strategies have not been established at the manufacturing facilities. We aimed to evaluate the safety and effects of continuous exposure to low-dose chlorine dioxide (ClO2) gas on cell fates, and to determine the optimum dose for safe usage of this disinfectant. Methods: We cultured an iPS cell line in the absence or presence of various doses of ClO2 gas. We evaluated cell proliferation, cell death, the maintenance of undifferentiated state, and cell senescence. Results: We found that iPS cell proliferation was not affected by 0.05 or 0.1 ppmv ClO2 gas in the atmosphere. Although 0.1 ppmv ClO2 slightly affected apoptosis, it was not a significant effect. Moreover, neither at 0.05 nor 0.1 ppmv ClO2 gas significantly affected the characteristics of iPS cells. Discussion and conclusion: Continuous exposure to 0.05 or 0.1 ppmv ClO2 gas did not affect the fate of iPS cells. These results may contribute to the development of new strategies for hygiene control in cell processing facilities.

Effects of continuous exposure to low concentration of ClO2 gas on the growth, viability, and maintenance of undifferentiated MSCs in long-term cultures.

INTRODUCTION: Hygienic management is more important in the manufacturing of cell products than in the production of chemical agents, because cell material and final product cannot be decontaminated. On the other hand, especially in the selection of hygienic agent, the adverse effects on the cells must be considered as well as the decontamination effect. ClO2 is a potent disinfectant, which is now expected as a safe and effective hygienic agent in the field of cell production. In this study, we investigated the effects of low dose ClO2 gas in the atmosphere of CO2 incubator on the characteristics of MSCs cultured in it. METHODS: First, we installed a ClO2 generator to a CO2 incubator for cell culture in which a constant level of ClO2 can be maintained. After culturing human cord derived MSCs in the CO2 incubator, the characteristics of cells were analyzed. RESULTS: Continuous exposure to 0.05 ppmv of ClO2 gas did not affect cell proliferation until at least 8th passage. In the FACS analysis, antigens usually expressed on MSCs, CD105, CD90, CD44, CD73 and CD29, were positively observed, but differentiation markers, CD11b and CD34, were little expressed on the MSCs exposed to 0.05 ppmv or 0.1 ppmv of ClO2 gas just as on the control cells. Also in the investigation for cell death, 0.05 ppmv and 0.1 ppmv of ClO2 gas little affected the viability, apoptosis or necrosis of MSCs. Furthermore, we assessed senescence using SA-ß-gal staining. Although the frequency of stained cells cultured in 0.1 ppmv of ClO2 gas was significantly increased than that of not exposed cells, the stained cells in 0.05 ppmv were rare and their frequency was almost the same as that in control. CONCLUSIONS: All these results indicate that, although excessive concentration of ClO2 gas induces senescence but neither apoptosis nor cell differentiation, exposure to 0.05 ppmv of ClO2 gas little affected the characteristics of MSCs. In this study we demonstrate that continuous exposure to appropriate dose of ClO2 gas can be safely used as decontamination agent in cell processing facilities.

Randomized, controlled trial of secondary prevention of coronary sclerosis in normocholesterolemic patients using pravastatin: final 5-year angiographic follow-up of the Prevention of Coronary Sclerosis (PCS) study.

BACKGROUND: The potential benefit of cholesterol-lowering therapy for normocholesterolemic patients with coronary artery disease (CAD) has not been clarified. The Prevention of Coronary Sclerosis (PCS) Study was designed to evaluate the effect of pravastatin on secondary prevention of progression of CAD in normocholesterolemic patients for a period of 5 years. METHODS: A total of 329 patients with CAD were enrolled. Normocholesterolemic patients were defined by a serum total cholesterol (TC) level of 180-219 mg/dl. Patients in this group were randomized into pravastatin and dietary control groups. Patients whose serum TC level fell outside the normal range were divided into a high-cholesterol reference group (TC > or = 220 mg/dl) and a low-cholesterol reference group (TC < 180 mg/dl). Patients in the pravastatin and high-cholesterol groups received pravastatin 10 mg/day. Coronary angiography was performed at baseline, 2 years, and 5 years and analyzed by quantitative coronary arteriography. Angiographic coronary progression was evaluated by minimum obstruction diameter (MOD) and mean segment diameter (MSD). RESULTS: At 5 years, change in MOD was significantly (change, P=0.033; change/years, P=0.032) less in the pravastatin group (-0.04 +/- 0.17 mm) than in the dietary control group (-0.16 +/- 0.27 mm). Although a similar trend was observed in the MSD results, the differences were not significant. CONCLUSION: Long-term angiographic data show that cholesterol-lowering therapy by pravastatin prevents progression of coronary atherosclerosis in normocholesterolemic patients with CAD.

Clinical outcome of percutaneous coronary intervention with antecedent mutant t-PA administration for acute myocardial infarction.

OBJECTIVE: We investigated the acute-phrase and chronic-phase outcomes of patients with acute myocardial infarction (AMI) undergoing percutaneous coronary intervention (PCI) with or without antecedent mutant tissue-type plasminogen (t-PA) administration. METHODS: Thirty-nine patients with a first AMI within 6 hours of onset were randomly assigned to the treatment group (1,600,000 IU IV monteplase, n = 19) or the nontreatment group (n = 20), followed by PCI. Clinical outcomes were then evaluated. RESULTS: Patient characteristics did not differ between the 2 groups. A significantly higher number of patients in the monteplase group achieved Thrombolysis In Myocardial Infarction trial (TIMI) grade 2 flow or more at the first angiography (84.2% vs 40.0%; P <.005), reduced number of devices (1.44 vs 1.80 devices, P <.05), and reduced procedure times (59.7 vs 86.7 minutes; P <.01), with no differences in peak creatine kinase and rates of major complications and no reflow or distal embolization. Observation over an average of 5.5 months revealed a tendency toward lower target lesion revascularization rates in the monteplase group (17.6% vs 31.6%) but no intergroup difference in rates of major complications. Pretreatment quantitative coronary angioplasty only showed a significant difference in minimal lumen diameter and percent diameter stenosis in the acute phase (1.13 mm in the monteplase group vs 0.66 mm in the nontreatment group, 57.0% vs 73.0%; P <.05). (99m)Tc-QGS (quantitative electrocardiographically gated single-photon emission computed tomographic scintigraphy) showed no intergroup differences in left ventricular end-diastolic volume index, end- systolic volume index, and ejection fraction in the acute and chronic phases. CONCLUSIONS: Our results suggest that PCI with antecedent mutant t-PA for AMI not only accelerates reperfusion, thereby facilitating PCI, but also attenuates restenosis in the chronic phase.

Effect of preintervention remodeling type on subsequent coronary artery behavior after directional atherectomy.

To evaluate the influence of preintervention remodeling on subsequent vessel behavior after directional coronary atherectomy (DCA) under intravascular ultrasound (IVUS) guidance, serial (before and after DCA and at 6-month follow-up) IVUS data were analyzed for 246 lesions that were classified into 2 categories: positive remodeling (PR) in 77 lesions versus intermediate or negative remodeling in 169 lesions. Although the 2 groups had similar baseline characteristics, IVUS data showed that the PR group had a greater acute lumen area (LA) gain without an increased late LA loss, resulting in a greater net (acute plus late) LA gain and follow-up LA. This suggests that IVUS-guided DCA may neutralize the negative impact of preintervention PR on late vessel patency.

Percutaneous transluminal septal myocardial ablation for hypertrophic obstructive cardiomyopathy.

Percutaneous transluminal septal myocardial ablation (PTSMA) is a new therapeutic option for patients with hypertrophic obstructive cardiomyopathy (HOCM). In the present study, the acute and follow-up results of PTSMA were evaluated. From August 1997 to March 2003 27 medically refractory patients (New York Heart Association (NYHA) functional class 2.9+/-0.6) with HOCM underwent PTSMA. The target septal branch was determined by probationary ballooning in 3 and by myocardial contrast echocardiography in 24 patients. The mean resting left ventricular outflow tract pressure gradient (PG) was reduced from 70+/-44 to 24+/-22 mmHg (p<0.0001); the peak concentration of creatine kinase was 1545+/-686 IU/L. Although transient trifascicular block was observed in 14 patients, permanent pacemaker implantation was not required. There were no major adverse cardiac events during the hospital stay; the mean clinical follow-up was 2.2+/-1.7 years. Repeated PTSMA was needed in 1 patient; however, symptomatic improvement had been well preserved in all patients (NYHA class 1.2+/-0.4). Follow-up echocardiographic examination showed sustained improvement in PG, septal and left ventricular posterior wall thicknesses, and the grade of systolic anterior movement and regurgitation of the mitral valve. In conclusion, PTSMA is a safe and effective therapeutic option for medically refractory patients with HOCM.

Aortic valve replacement for a patient with porcelain aorta and retrosternal gastric tube reconstruction after esophageal resection.

A 67-year-old woman with congestive heart failure due to aortic stenosis and regurgitation needed aortic valve replacement. She had undergone right radical mastectomy 23 years before, and total thoracic esophagectomy with retrosternal gastric tube reconstruction 11 years before. Plain computed tomography showed coincident porcelain aorta. Aortic valve replacement was performed through a median sternotomy approach. Blunt dissection on anterior and right side of the gastric tube could be done with minimal injury, and the heart was exposed as in usual cardiac surgery. Preoperative multi-detector computed tomography revealed inhomogeneous and patchy distribution of calcification in the ascending aorta, and was helpful to decide aortotomy site. Aortic valve replacement was done and aortotomy was closed with felt strip buttressed running suture. Postoperative course was uneventful.