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Quantification of Basal and Stimulated ROS Levels as Predictors of Islet Potency and Function 总被引:2,自引:0,他引:2
B. Armann M. S. Hanson E. Hatch A. Steffen L. A. Fernandez 《American journal of transplantation》2007,7(1):38-47
We have developed a luminol-based assay using intact islets, which allows for quantification of reactive oxygen species (ROS). In addition, an index capable of characterizing metabolic and mitochondrial integrity prior to transplantation was created based on the capacity of islets to respond to high glucose and rotenone (mitochondrial respiratory chain complex I inhibitor) by production of ROS. To validate this assay, lipid peroxidation and antioxidative defense capacity were evaluated by detection of malondialdehyde (MDA) levels and glutathione peroxidase activity (GPx), respectively. Also, flow cytometric analyses of ROS (dihydroethidine), apoptosis (Annexin V, active caspases), necrosis (Topro3), and mitochondrial membrane potential (JC-1) were done in parallel to correlate with changes in luminol-measured ROS. ATP/ADP ratios were quantified by HPLC and the predictive value of ROS measurement on islet functional potency was correlated with capacity to reverse diabetes in a streptozotocin-induced diabetic NOD.scid mouse model as well as in human transplant recipients. Our data demonstrate that levels of ROS in islets correlate with the percentage of apoptotic cells and their functional potency in vivo. The ROS indices following glucose and rotenone exposure are indicative of metabolic potency and mitochondrial integrity and can be used as surrogate markers to evaluate the quality of islets prior to transplantation. 相似文献
33.
Antonio Gigante Claudia Bevilacqua Andrea Ricevuto Monica Mattioli-Belmonte Francesco Greco 《Knee surgery, sports traumatology, arthroscopy》2007,15(1):88-92
The implantation of chondrocytes, seeded on matrices such as hyaluronic acid or collagen membranes, is a method that is being widely used for the treatment of chondral defects. The aim of the present study was to evaluate the distribution, viability and phenotype expression of the cells seeded on a collagen membrane just at the time of the implantation. Twelve patients who were suffering from articular cartilage lesions were treated by the MACI® procedure. The residual part of each membrane was tested by colorimetric assay (MTT) and histochemical and ultrastructural analyses were carried out. In all of the samples a large number of viable cells, quite homogenously distributed, was detected. The cells expressed the markers of the differentiated hyaline chondrocytes. These data reassure in that the MACI procedure provides a suitable engineered tissue for cartilage repair, in line with the clinical evidences emerging in the literature. 相似文献
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Monica A McHenry 《J. Speech Lang. Hear. Res.》2003,46(3):702-710
There are many potential sources of variability in speech production, particularly in individuals with dysarthria. The degree and time course of stabilization of the speech production system during recovery from a neurological insult is not constant across individuals. Another source of variability in speech production is speaking rate. Although individuals with no neurological impairments typically show increased variability at reduced speaking rates, this phenomenon has not been explored extensively in individuals with dysarthria. Because rate control strategies are commonly used in dysarthria treatment, it is of clinical importance to know if individuals with dysarthria produce less variable speech with rate reduction. Six individuals with mild dysarthria, 6 with moderate-to-severe dysarthria, and 6 matched normal controls repeated an utterance in four speaking rate conditions: habitual, fast, breaks between words, and stretched. Data were analyzed using the spatiotemporal index (STI), a composite measure of spatial and temporal variability across token repetitions. The normal controls consistently demonstrated the least variability, regardless of rate condition. Both groups with dysarthria were the least variable in the stretched condition and the most variable in the fast condition. The STI values of the group with moderate-to-severe dysarthria were significantly different from both the individuals with mild dysarthria and the normal controls. There were no significant differences between the group with mild dysarthria and the normal controls. In general, slowing the speaking rate in individuals with dysarthria reduces spatiotemporal variability; however, the effect of reduced spatiotemporal variability on intelligibility requires further investigation. 相似文献
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Monica Conciatori Christopher J Stodgell Susan L Hyman Melanie O'Bara Roberto Militerni Carmela Bravaccio Simona Trillo Francesco Montecchi Cindy Schneider Raun Melmed Maurizio Elia Lori Crawford Sarah J Spence Lucianna Muscarella Vito Guarnieri Leonardo D'Agruma Alessandro Quattrone Leopoldo Zelante Daniel Rabinowitz Tiziana Pascucci Stefano Puglisi-Allegra Karl-Ludvig Reichelt Patricia M Rodier Antonio M Persico 《Neuropsychopharmacology》2004,55(4):413-419
BACKGROUND: The HOXA1 gene plays a major role in brainstem and cranial morphogenesis. The G allele of the HOXA1 A218G polymorphism has been previously found associated with autism. METHODS: We performed case-control and family-based association analyses, contrasting 127 autistic patients with 174 ethnically matched controls, and assessing for allelic transmission disequilibrium in 189 complete trios. RESULTS: A, and not G, alleles were associated with autism using both case-control (chi(2) = 8.96 and 5.71, 1 df, p <.005 and <.025 for genotypes and alleles, respectively), and family-based (transmission/disequilibrium test chi(2) = 8.80, 1 df, p <.005) association analyses. The head circumference of 31 patients carrying one or two copies of the G allele displayed significantly larger median values (95.0th vs. 82.5th percentile, p <.05) and dramatically reduced interindividual variability (p <.0001), compared with 166 patients carrying the A/A genotype. CONCLUSIONS: The HOXA1 A218G polymorphism explains approximately 5% of the variance in the head circumference of autistic patients and represents to our knowledge the first known gene variant providing sizable contributions to cranial morphology. The disease specificity of this finding is currently being investigated. Nonreplications in genetic linkage/association studies could partly stem from the dyshomogeneous distribution of an endophenotype morphologically defined by cranial circumference. 相似文献
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Roberto Banfi Eleonora Pavone Rita Cerri Monica Menichetti 《Pharmacy World & Science》2007,29(4):400-403
In Italy mifepristone is not yet marketed. Gynaecologists in our hospital asked to use this medication as a less traumatic
method for voluntary abortions. We followed the standard procedure defined by the Italian Health Ministry (IMH) for purchasing
drugs from abroad but encountered several unexpected barriers. Starting from this case, this paper is aimed at identifying
these barriers which we found to be not only professional, but also ethical, religious and moral. 相似文献