51 research outputs found
Mitochondrial DNA D-loop variants correlate with a primary open-angle glaucoma subgroup
IntroductionPrimary open-angle glaucoma (POAG) is a characteristic optic neuropathy, caused by degeneration of the optic nerve-forming neurons, the retinal ganglion cells (RGCs). High intraocular pressure (IOP) and aging have been identified as major risk factors; yet the POAG pathophysiology is not fully understood. Since RGCs have high energy requirements, mitochondrial dysfunction may put the survivability of RGCs at risk. We explored in buffy coat DNA whether mtDNA variants and their distribution throughout the mtDNA could be risk factors for POAG.MethodsThe mtDNA was sequenced from age- and sex-matched study groups, being high tension glaucoma (HTG, n=71), normal tension glaucoma patients (NTG, n=33), ocular hypertensive subjects (OH, n=7), and cataract controls (without glaucoma; n=30), all without remarkable comorbidities.ResultsNo association was found between the number of mtDNA variants in genes encoding proteins, tRNAs, rRNAs, and in non-coding regions in the different study groups. Next, variants that controls shared with the other groups were discarded. A significantly higher number of exclusive variants was observed in the D-loop region for the HTG group (~1.23 variants/subject), in contrast to controls (~0.35 variants/subject). In the D-loop, specifically in the 7S DNA sub-region within the Hypervariable region 1 (HV1), we found that 42% of the HTG and 27% of the NTG subjects presented variants, while this was only 14% for the controls and OH subjects. As we have previously reported a reduction in mtDNA copy number in HTG, we analysed if specific D-loop variants could explain this. While the majority of glaucoma patients with the exclusive D-loop variants m.72T>C, m.16163 A>G, m.16186C>T, m.16298T>C, and m.16390G>A presented a mtDNA copy number below controls median, no significant association between these variants and low copy number was found and their possible negative role in mtDNA replication remains uncertain. Approximately 38% of the HTG patients with reduced copy number did not carry any exclusive D-loop or other mtDNA variants, which indicates that variants in nuclear-encoded mitochondrial genes, environmental factors, or aging might be involved in those cases.ConclusionIn conclusion, we found that variants in the D-loop region may be a risk factor in a subgroup of POAG, possibly by affecting mtDNA replication
Preparation of a polyurethane scaffold for tissue engineering made by a combination of salt leaching and freeze-drying of dioxane
Molecular alterations induced by a high-fat high-fiber diet in porcine adipose tissues: variations according to the anatomical fat location
Fusion of Wild-Type Mesoangioblasts with Myotubes of mtDNA Mutation Carriers Leads to a Proportional Reduction in mtDNA Mutation Load
In 25% of patients with mitochondrial myopathies, pathogenic mitochondrial DNA (mtDNA) mutation are the cause. For heteroplasmic mtDNA mutations, symptoms manifest when the mutation load exceeds a tissue-specific threshold. Therefore, lowering the mutation load is expected to ameliorate disease manifestations. This can be achieved by fusing wild-type mesoangioblasts with mtDNA mutant myotubes. We have tested this in vitro for female carriers of the m.3271T>C or m.3291T>C mutation (mutation load >90%) using wild-type male mesoangioblasts. Individual fused myotubes were collected by a newly-developed laser capture microdissection (LCM) protocol, visualized by immunostaining using an anti-myosin antibody. Fusion rates were determined based on male-female nuclei ratios by fluorescently labelling the Y-chromosome. Using combined ‘wet’ and ‘air dried’ LCM imaging improved fluorescence imaging quality and cell yield. Wild-type mesoangioblasts fused in different ratios with myotubes containing either the m.3271T>C or the m.3291T>C mutation. This resulted in the reduction of the mtDNA mutation load proportional to the number of fused wild-type mesoangioblasts for both mtDNA mutations. The proportional reduction in mtDNA mutation load in vitro after fusion is promising in the context of muscle stem cell therapy for mtDNA mutation carriers in vivo, in which we propose the same strategy using autologous wild-type mesoangioblasts
Blood biomarkers for assessment of mitochondrial dysfunction:An expert review
Although mitochondrial dysfunction is the known cause of primary mitochondrial disease, mitochondrial dysfunction is often difficult to measure and prove, especially when biopsies of affected tissue are not available. In order to identify blood biomarkers of mitochondrial dysfunction, we reviewed studies that measured blood biomarkers in genetically, clinically or biochemically confirmed primary mitochondrial disease patients. In this way, we were certain that there was an underlying mitochondrial dysfunction which could validate the biomarker. We found biomarkers of three classes: 1) functional markers measured in blood cells, 2) biochemical markers of serum/plasma and 3) DNA markers. While none of the reviewed single biomarkers may perfectly reveal all underlying mitochondrial dysfunction, combining biomarkers that cover different aspects of mitochondrial impairment probably is a good strategy. This biomarker panel may assist in the diagnosis of primary mitochondrial disease patients. As mitochondrial dysfunction may also play a significant role in the pathophysiology of multifactorial disorders such as Alzheimer's disease and glaucoma, the panel may serve to assess mitochondrial dysfunction in complex multifactorial diseases as well and enable selection of patients who could benefit from therapies targeting mitochondria
Human embryonic stem cell-derived cardiomyocytes as an in vitro model to study cardiac insulin resistance
Displacement of the medial meniscus within the passive motion characteristics of the human knee joint: an RSA study in human cadaver knees.
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47520.pdf (publisher's version ) (Closed access)The objective of this study was to validate an in vitro human cadaver knee-joint model for the evaluation of the meniscal movement during knee-joint flexion. The question was whether our model showed comparable meniscal displacements to those found in earlier meniscal movement studies in vivo. Furthermore, we determined the influence of tibial torque on the meniscal displacement during knee-joint flexion. Three tantalum beads were inserted in the medial meniscus of six human-cadaver joints. The knee joints were placed and loaded in a loading apparatus, and the movements of the beads were determined by means of RSA during knee-joint flexion and extension with and without internal tibial (IT) and external tibial (ET) torque. During flexion without tibial torque, all menisci moved in posterior and lateral direction. The anterior horn showed significantly greater excursions than the posterior horn in both posterior and lateral direction. Internal tibial torque caused an anterior displacement of the pathway on the tibial plateau. External tibial torque caused a posterior displacement of the pathway. External tibial torque restricted the meniscal displacement during the first 30 degrees of knee-joint flexion. The displacements of the meniscus in this experiment were similar to the displacements described in the in vivo MRI studies. Furthermore, the application of tibial torque confirmed the relative immobility of the posterior horn of the meniscus. During external tibial torque, the posterior displacement of the pathway on the tibial plateau during the first 30 degrees of flexion might be restricted by the attached knee-joint capsule or the femoral condyle. This model revealed representative meniscal displacements during simple knee-joint flexion and also during the outer limits of passive knee-joint motion
Low mitochondrial DNA copy number in buffy coat DNA of primary open-angle glaucoma patients
Preparation of a polyurethane scaffold for tissue engineering made by a combination of salt leaching and freeze-drying of dioxane
In the past several types of synthetic porous materials have been made as meniscus reconstruction materials. Most of these materials lacked a good combination between suitable mechanical properties and a high interconnectivity. In this work porous scaffolds were made from the polyurethane Estane 5701-F1 by freeze drying of a dioxane and water solution in combination with salt leaching. It was possible to obtain very porous scaffolds with a very high interconnectivity. Porosity, pore size and interconnectivity can be independently adjusted by varying the amount of water, porogen size and the amount of porogen used. The obtained compression moduli of the scaffolds were between 40 kPa and 400 kPa with a variation in porosity between 72 and 87%. These scaffolds are very suitable for the use as meniscus replacement materials.
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