N1-methylation of adenosine (m1A) in ND5 mRNA leads to complex I dysfunction in Alzheimer's disease.

One mechanism of particular interest to regulate mRNA fate post-transcriptionally is mRNA modification. Especially the extent of m1A mRNA methylation is highly discussed due to methodological differences. However, one single m1A site in mitochondrial ND5 mRNA was unanimously reported by different groups. ND5 is a subunit of complex I of the respiratory chain. It is considered essential for the coupling of oxidation and proton transport. Here we demonstrate that this m1A site might be involved in the pathophysiology of Alzheimer's disease (AD). One of the pathological hallmarks of this neurodegenerative disease is mitochondrial dysfunction, mainly induced by Amyloid ß (Aß). Aß mainly disturbs functions of complex I and IV of the respiratory chain. However, the molecular mechanism of complex I dysfunction is still not fully understood. We found enhanced m1A methylation of ND5 mRNA in an AD cell model as well as in AD patients. Formation of this m1A methylation is catalyzed by increased TRMT10C protein levels, leading to translation repression of ND5. As a consequence, here demonstrated for the first time, TRMT10C induced m1A methylation of ND5 mRNA leads to mitochondrial dysfunction. Our findings suggest that this newly identified mechanism might be involved in Aß-induced mitochondrial dysfunction.

Effect of three-dimensional expansion and cell seeding density on the cartilage-forming capacity of human articular chondrocytes in type II collagen sponges.

Chondrocytes for tissue engineering strategies are typically expanded in monolayer (2D), leading to cell dedifferentiation but allowing to generate large cell numbers for seeding into scaffolds. Direct chondrocyte culture in scaffolds, instead, may support better maintenance of the differentiated phenotype but reduce the extent of proliferation and thus the resulting cell density. This study investigates whether the quality of cartilaginous tissues generated in vitro by human articular chondrocytes (HAC) on type II collagen sponges is enhanced (1) by direct expansion on the scaffolds (3D), as compared with standard 2D, or (2) by increasing cell seeding density, which in turn requires extensive 2D expansion. Three-dimensional expansion of HAC on the scaffolds, as compared with 2D expansion for the same number of doublings, better maintained the chondrocytic phenotype of the expanded cells (13.7-fold higher levels of type II collagen mRNA) but did not enhance their accumulation of glycosaminoglycan (GAG) following chondrogenic culture. Instead, increasing the HAC seeding density in the scaffolds (from 25 × 10(3) to 66 × 10(3) cells/mm(3)) significantly improved chondrogenesis (up to 3.3-fold higher GAG accumulation and up to 9.3-fold higher type II collagen mRNA), even if seeded cells had to be expanded and dedifferentiated more extensively in 2D to reach the required cell numbers. This study indicates that, under the specific conditions tested, a high-seeding density of HAC in 3D scaffolds is more critical for the generation of cartilaginous constructs than the stage of cell differentiation reached following expansion.

Chondrogenesis of human mesenchymal stem cells by local transforming growth factor-beta delivery in a biphasic resorbable carrier.

Little is known about the potential of growth factor-augmented biphasic implants composed of a gel and a solid scaffold to enhance chondrogenesis of mesenchymal stem cells (MSCs). We analyzed whether a collagen type I/III carrier and fibrin glue (FG) combined to a biphasic construct support in vitro chondrogenesis of MSCs and allow for local release of bioactive transforming growth factor-beta1 (TGF-beta1). Further, a possible advantage of partial autologous fibrin glue (PAF) over commercial FG was assessed. Collagen carriers seeded with 5 x 10(5) human MSCs with or without FG, PAF, or TGF-beta1-upgraded FG were cultured for 6 weeks in chondrogenic medium with or without TGF-beta1. Pellets with or without FG/PAF served as controls. FG and collagen carriers allowed strong upregulation of COL2A1, AGC, and COL10A1 mRNA, deposition of collagen-type II, and mediated a significantly higher proteoglycan content compared with biomaterial-free pellets. Collagen-carrier groups contained significantly more proteoglycan than FG and PAF pellets, whereas biphasic PAF-carrier constructs were inferior to FG-carrier constructs. Upgrading of biphasic FG-carrier constructs with 50 ng TGF-beta1/construct mediated chondrogenesis as successfully as supply of TGF-beta1 via the medium. In conclusion, the biphasic carrier constructs showed a high biofunctionality by continuous form stability with improved chondrogenesis and long-term local supply of bioactive TGF-beta1 which may be useful to enhance matrix-assisted repair strategies for damaged cartilage.

Design and technical adjustment of a water treatment system: 15 years of experience.

This article gives a technical summary over the development of our dialysis water preparation site covering more than a decade of common effort to improve and secure water quality. We invite the reader to follow us through the experience of 3 major conception changes determined by available localities and resources, operational difficulties, and maintenance as well as progressive technologies and standards. Critical analysis aims to provide information to promote development in this area including details of preventable errors and misconceptions. The findings show that it is possible to achieve higher water quality on regular bases at affordable cost through technical changes, often minor; preventive maintenance; and continual monitoring of all components.

New nitrogen mustards structurally related to (L)-carnitine.

Enantiopure nitrogen mustards which mimic (L)-carnitine framework are prepared by a multi-step synthesis from the (R)-di-tert-butyl malate and their antitumor properties evaluated.

Effective flow performances and dialysis doses delivered with permanent catheters: a 24-month comparative study of permanent catheters versus arterio-venous vascular accesses.

BACKGROUND: Permanent venous catheters have emerged as a long-term vascular access option for renal replacement therapy in end-stage renal disease patients. The design and venous location of catheter devices bear intrinsic flow limitations that may negatively affect the adequacy of dialysis and the patient outcome. There is limited data comparing the long-term dialysis adequacy delivered with permanent catheters vs arterio-venous vascular accesses (AVA). METHODS: To explore this problem, we conducted a prospective 24-month trial comparing the flow performances and dialysis dose (Kt/Vdp) deliveries of both access options in a group of 42 haemodialysis patients during two study phases. During the first 12 months the patients completed a treatment period by means of permanent dual silicone catheters (DualKT). Then they were transferred to an AVA (40 native arterio-venous fistulas and two PTFE grafts) and monitored for an additional 12-month period. Assessments of flow adequacy and dialysis quantification were performed monthly. RESULTS: Dialysis adequacy was achieved in all cases. No patient had to be transferred prematurely to the AVA because of catheter failure. Three catheters had to be replaced due to bacteraemia in three patients. The mean effective blood flow rates achieved were 316+/-3.5 ml/min and 340+/-3.3 ml/min with DualKT and AVA, respectively, for a pre-set machine blood flow of 348+/-2.2 ml/min. Recirculation rates evaluated with the 'slow blood flow' method were 8.6+/-0.6 and 12.1+/-0.8% for DualKT and AVA using mean values of the solute markers urea and creatinine. Due to the possibility of a comparison veno-venous vs arterio-venous blood circulation, a corrected arterio-venous access recirculation could be derived from the difference between the two, which was around 3%. The blood flow resistance of the DualKT was slightly higher than with AVA as indicated by venous pressure differences. Kt/Vdp delivered was 1.37+/-0.02 and 1.45+/-0.02 with DualKT and AVA access respectively. The loss of dialysis efficacy using catheters was estimated at 6%. However, in all cases Kt/Vdp values remained above the recommended values (Kt/Vdp > or = 1.2). Protein nutritional state, as well as conventional clinical and biochemical markers of dialysis adequacy, remained in the optimal range. CONCLUSION: Permanent venous catheters provide adequate haemodialysis on a long-term basis. Flow performances and dialysis doses are slightly reduced (5-6%) when compared with AVA. Regular assessment of dialysis performance is strongly recommended to assure dialysis adequacy. Lengthening dialysis time may represent a simple and efficient tool to compensate for reduced flow performances with catheter use.