Hole burning imaging studies of cancerous and analogous normal ovarian tissues utilizing organelle specific dyes

In order to elucidate differences between cancerous and analogous normal tissues, nonphotochemical hole burning (NPHB) spectroscopy utilizing MF680 (commonly known as rhodamine 800) was performed. The cationic lipophilic fluorophore MF680 preferentially locates in in situ mitochondria due to the presence of a large negative membrane potential (relative to the cellular cytoplasm) caused by oxidative phosphorylation. The cancerous ovarian (adenocarcinoma, stage 3C) and normal peritoneal tissue samples were surgically removed and characterized at the University of Oklahoma-Health Science Center (Oklahoma City, OK), where the normal tissues were then further categorized as adequate or distant normal, depending on the physical proximity to the cancerous tissue.;The results of these hole burning experiments are interpreted on the basis of the NPHB mechanism and characteristic interactions between the glass-like structured host (in vivo mitochondrial matrix) and the guest (MF680) in the burning of spectral holes, thus providing an image of the cellular ultrastructure. The fluorescence excitation spectra of MF680 in the tissues and the confocal microscopy images of thin sliced tissues incubated with MF680 confirmed the site-specificity of the probe molecules in the cellular systems. A direct comparison of the positions of the MF680 fluorescence excitation peaks between tissues showed only insignificant differences. Hole growth kinetics of MF680 in tissues, unlike the preliminary cultured cell studies, also proved inconclusive in distinguishing the tissue cellular matrices surrounding the chromophores. Measurement of changes in the permanent dipole moment ( f·Deltamu) was accomplished by measurement of changes in hole width in response to the application of an external electric (Stark) field. Stark fields resulted in hole broadening for both tissue samples when laser polarization was either parallel or perpendicular to the applied electric field. The f·Deltamu values for the cancerous tissues were found to be ~1.35 times greater than those of the distant normal analogs on all burn frequencies. The differences in the change in the dipole moments are interpreted in terms of effects from the mitochondrial membrane potential. Results for flow cytometric determination of the mitochondrial membrane potential in the attempt to quantificate f·Deltamu values are also presented

Dimethyl sulfoxide-respiring bacteria in Suribati Ike, a hypersaline lake, in Antarctica and the marine environment

Dimethyl sulfoxide (DMSO) occurs worldwide, especially in marine environments as well as in lakes and rainwater. DMSO respiration by bacteria is assumed to play an important role in the sulfur cycle in Antarctica and on earth. We first studied whether DMSO-respiring bacteria existed in Antarctica. Eight strains were isolated that grew by DMSO respiration under anaerobic conditions from water of the halocline in a meromictic lake, Suribati Ike, near Syowa Station in Antarctica. All of them were related to known species belonging to the genus Marinobacter based on 16S rRNA gene sequences. Using a clone library analysis of 16S rRNA gene sequences, 38 of total 48 clones from water of the halocline were identified as Marinobacter. Studies on the various anaerobic respiration capabilities by bacteria in the halocline water found only DMSO respiration. Studies on bacteria with anaerobic respiration abilities in seawater from the Pacific Ocean and Seto Inland Sea, showed that either DMSO-respiring or nitrate-respiring bacteria were present and that all of isolates capable of DMSO respiration were closely related to Vibrio species

Validity and reproducibility of folate and vitamin B12 intakes estimated from a self-administered diet history questionnaire in Japanese pregnant women

<p>Abstract</p> <p>Background</p> <p>No validated dietary questionnaire for assessing folate and vitamin B₁₂intakes during pregnancy is available in Japan. We evaluated the validity and reproducibility of intakes of folate and vitamin B₁₂estimated from a self-administered diet history questionnaire (DHQ) in Japanese pregnant women.</p> <p>Methods</p> <p>A sample of 167 healthy subjects with singleton pregnancies in the second trimester was recruited at a private obstetric hospital in metropolitan Tokyo from June to October 2008 (n = 76), and at a university hospital in Tokyo from June 2010 to June 2011 (n = 91). The dietary intakes of folate and vitamin B₁₂were assessed using the DHQ. The serum concentrations of folate and vitamin B₁₂were measured as reference values in the validation study. To assess the reproducibility of the results, 58 pregnant women completed the DHQ twice within 4-5 week interval.</p> <p>Results</p> <p>Significantly positive correlations were found between energy-adjusted intakes and serum concentrations of folate and vitamin B₁₂(r = 0.286, <it>p </it>< 0.001 and r = 0.222, <it>p </it>= 0.004, respectively). After excluding the participants with nausea (n = 121), the correlation coefficient for vitamin B₁₂increased to 0.313 (<it>p </it>= 0.001). When participants were classified into quintiles based on intakes and serum concentrations of folate and vitamin B₁₂, approximately 60% were classified in the same or adjacent quintile. The intraclass correlation coefficients of the two-time DHQ were 0.725 for folate and 0.512 for vitamin B₁₂.</p> <p>Conclusion</p> <p>The present study indicated that the DHQ had acceptable validity and reproducibility for assessing folate and vitamin B₁₂intakes in Japanese pregnant women.</p

Beneficial effect of tetrahydrobiopterin on ischemia-reperfusion injury in isolated perfused rat hearts

AbstractObjective: It has recently been proposed that nitric oxide synthase, in the presence of suboptimal levels of tetrahydrobiopterin, an essential cofactor of this enzyme, might favor increased production of oxygen radicals. The aim of this study was to clarify whether supplement with tetrahydrobiopterin would exert a cardioprotective effect against ischemia-reperfusion injury. Methods: Isolated perfused rat hearts were subjected to 30 minutes of global ischemia and 30 minutes of reperfusion at 37°C. Hearts were treated with tetrahydrobiopterin or vehicle for 5 minutes just before ischemia and during the first 5 minutes of the reperfusion period. Effects of tetrahydrobiopterin on left ventricular function, myocardial contents of lipid peroxidation and high-energy phosphates, and levels of lactate dehydrogenase and nitrite plus nitrate in perfusate during ischemia and after reperfusion were estimated and further compared with those of superoxide dismutase plus catalase or l-ascorbic acid. Results: Tetrahydrobiopterin and superoxide dismutase plus catalase both improved contractile and metabolic abnormalities in reperfused hearts. On the other hand, l-ascorbic acid at a dose having an equipotent radical scavenging activity with tetrahydrobiopterin did not significantly affect the postischemic changes. Although tetrahydrobiopterin and superoxide dismutase plus catalase significantly alleviated ischemic contracture during ischemia, diminished perfusate levels of nitrite plus nitrate after reperfusion were restored only with tetrahydrobiopterin. Conclusion: Results demonstrated that tetrahydrobiopterin lessens ischemia-reperfusion injury in isolated perfused rat hearts, probably independent of its intrinsic radical scavenging action. The cardioprotective effect of tetrahydrobiopterin implies that tetrahydrobiopterin could be a novel and effective therapeutic option in the treatment of ischemia-reperfusion injury.J Thorac Cardiovasc Surg 2002;124:775-8

High HbA1c levels correlate with reduced plaque regression during statin treatment in patients with stable coronary artery disease: Results of the coronary atherosclerosis study measuring effects of rosuvastatin using intravascular ultrasound in Japanese subjects (COSMOS)

Abstract Background The incidence of cardiac events is higher in patients with diabetes than in people without diabetes. The Coronary Atherosclerosis Study Measuring Effects of Rosuvastatin Using Intravascular Ultrasound in Japanese Subjects (COSMOS) demonstrated significant plaque regression in Japanese patients with chronic coronary disease after 76 weeks of rosuvastatin (2.5 mg once daily, up-titrated to a maximum of 20 mg/day to achieve LDL cholesterol Methods In this subanalysis of COSMOS, we examined the association between HbA1c and plaque regression in 40 patients with HbA1c ≥6.5% (high group) and 86 patients with HbA1c Results In multivariate analyses, HbA1c and plaque volume at baseline were major determinants of plaque regression. LDL cholesterol decreased by 37% and 39% in the high and low groups, respectively, while HDL cholesterol increased by 16% and 22%, respectively. The reduction in plaque volume was significantly (p = 0.04) greater in the low group (from 71.0 ± 39.9 to 64.7 ± 34.7 mm3) than in the high group (from 74.3 ± 34.2 to 71.4 ± 32.3 mm3). Vessel volume increased in the high group but not in the low group (change from baseline: +4.2% vs −0.8%, p = 0.02). Change in plaque volume was significantly correlated with baseline HbA1c. Conclusions Despite similar improvements in lipid levels, plaque regression was less pronounced in patients with high HbA1c levels compared with those with low levels. Tight glucose control during statin therapy may enhance plaque regression in patients with stable coronary disease. Trial registration ClinicalTrials.gov, Identifier NCT00329160</p

Chronic Treatment with a Water-Soluble Extract from the Culture Medium of Ganoderma lucidum

Type 2 diabetes mellitus has been known to increase systemic oxidative stress by chronic hyperglycemia and visceral obesity and aggravate cerebral ischemic injury. On the basis of our previous study regarding a water-soluble extract from the culture medium of Ganoderma lucidum mycelia (designed as MAK), which exerts antioxidative and neuroprotective effects, the present study was conducted to evaluate the preventive effects of MAK on apoptosis and necroptosis (a programmed necrosis) induced by hypoxia/ischemia (H/I) in type 2 diabetic KKAy mice. H/I was induced by a combination of unilateral common carotid artery ligation with hypoxia (8% O2 for 20 min) and subsequent reoxygenation. Pretreatment with MAK (1 g/kg, p.o.) for a week significantly reduced H/I-induced neurological deficits and brain infarction volume assessed at 24 h of reoxygenation. Histochemical analysis showed that MAK significantly suppressed superoxide production, neuronal cell death, and vacuolation in the ischemic penumbra, which was accompanied by a decrease in the numbers of TUNEL- or cleaved caspase-3-positive cells. Furthermore, MAK decreased the expression of receptor-interacting protein kinase 3 mRNA and protein, a key molecule for necroptosis. These results suggest that MAK confers resistance to apoptotic and necroptotic cell death and relieves H/I-induced cerebral ischemic injury in type 2 diabetic mice

Examination of the Effective Utilization of the CARELINK® Remote Monitoring System after its Introduction

AbstractBackground: Japan started using the CARELINK® (Medtronic, Inc, Minneapolis, MN, USA) remote monitoring system in April 2009. However, in some cases, the device failed to transmit a message after registration or according to schedule. We investigated the difference between patients who could make effective use of CARELINK® system and those who could not.Subjects and Method: Sixty patients who had registered until December 2009 at our institution were analyzed. These patients were divided into two groups: those who were able to use the device effectively (group G, n = 49) and those who were not (group F = 11). Patient background, automatic or manual telemetries, new or existing implant patient, presence of adverse events, and the use or non-use of a checklist at the time of introduction were compared between the two groups.Results: In group G, more patients used a checklist at the time of introduction than that in group F (use of checklist/total, 31/49 in group G vs. 3/11 in group F; P, 0.029). No significant difference was observed in other factors between the two groups.Conclusion: We consider that the method used to explain the system are important to make the patients understand handling methods of CARELINK® system. The number of patients introduced to remote monitoring of implantable devices will continue to increase in the future; therefore, we must continue to develop innovative approaches for their effective us

Identification of a novel intronic enhancer responsible for the transcriptional regulation of musashi1 in neural stem/progenitor cells

<p>Abstract</p> <p>Background</p> <p>The specific genetic regulation of neural primordial cell determination is of great interest in stem cell biology. The Musashi1 (Msi1) protein, which belongs to an evolutionarily conserved family of RNA-binding proteins, is a marker for neural stem/progenitor cells (NS/PCs) in the embryonic and post-natal central nervous system (CNS). Msi1 regulates the translation of its downstream targets, including <it>m-Numb </it>and <it>p21 </it>mRNAs. <it>In vitro </it>experiments using knockout mice have shown that Msi1 and its isoform Musashi2 (Msi2) keep NS/PCs in an undifferentiated and proliferative state. Msi1 is expressed not only in NS/PCs, but also in other somatic stem cells and in tumours. Based on previous findings, Msi1 is likely to be a key regulator for maintaining the characteristics of self-renewing stem cells. However, the mechanisms regulating <it>Msi1 </it>expression are not yet clear.</p> <p>Results</p> <p>To identify the DNA region affecting <it>Msi1 </it>transcription, we inserted the fusion gene <it>ffLuc</it>, comprised of the fluorescent <it>Venus </it>protein and firefly <it>Luciferase</it>, at the translation initiation site of the mouse <it>Msi1 </it>gene locus contained in a 184-kb bacterial artificial chromosome (BAC). Fluorescence and Luciferase activity, reflecting the <it>Msi1 </it>transcriptional activity, were observed in a stable BAC-carrying embryonic stem cell line when it was induced toward neural lineage differentiation by retinoic acid treatment. When neuronal differentiation was induced in embryoid body (EB)-derived neurosphere cells, reporter signals were detected in Msi1-positive NSCs and GFAP-positive astrocytes, but not in MAP2-positive neurons. By introducing deletions into the BAC reporter gene and conducting further reporter experiments using a minimized enhancer region, we identified a region, "D5E2," that is responsible for <it>Msi1 </it>transcription in NS/PCs.</p> <p>Conclusions</p> <p>A regulatory element for <it>Msi1 </it>transcription in NS/PCs is located in the sixth intron of the <it>Msi1 </it>gene. The 595-bp D5E2 intronic enhancer can transactivate <it>Msi1 </it>gene expression with cell-type specificity markedly similar to the endogenous Msi1 expression patterns.</p