Regulation of hepatic cardiolipin metabolism by TNFα: Implication in cancer cachexia

International audienceCardiolipin (CL) content accumulation leads to an increase in energy wasting in liver mitochondria in a rat model of cancer cachexia in which tumor necrosis factor alpha (TNFα) is highly expressed. In this study we investigated the mechanisms involved in liver mitochondria CL accumulation in cancer cachexia and examined if TNFα was involved in this process leading to mitochondrial bioenergetics alterations. We studied gene, protein expression and activity of the main enzymes involved in CL metabolism in liver mitochondria from a rat model of cancer cachexia and in HepaRG hepatocyte-like cells exposed to 20 ng/ml of TNFα for 12 h. Phosphatidylglycerolphosphate synthase (PGPS) gene expression was increased 2.3-fold (p < 0.02) and cardiolipin synthase (CLS) activity decreased 44% (p < 0.03) in cachectic rat livers compared to controls. CL remodeling enzymes monolysocardiolipin acyltransferase (MLCL AT-1) activity and tafazzin (TAZ) gene expression were increased 30% (p < 0.01) and 50% (p < 0.02), respectively, in cachectic rat livers compared to controls. Incubation of hepatocytes with TNFα increased CL content 15% (p < 0.05), mitochondrial oxygen consumption 33% (p < 0.05), PGPS gene expression 44% (p < 0.05) and MLCL AT-1 activity 20% (p < 0.05) compared to controls. These above findings strongly suggest that in cancer cachexia, TNFα induces a higher energy wasting in liver mitochondria by increasing CL content via upregulation of PGPS expression

Small-molecule inhibition of MuRF1 attenuates skeletal muscle atrophy and dysfunction in cardiac cachexia

Background; Muscle ring finger 1 (MuRF1) is a muscle‐specific ubiquitin E3 ligase activated during clinical conditions associated with skeletal muscle wasting. Yet, there remains a paucity of therapeutic interventions that directly inhibit MuRF1 function, particularly in vivo. The current study, therefore, developed a novel compound targeting the central coiled coil domain of MuRF1 to inhibit muscle wasting in cardiac cachexia. Methods; We identified small molecules that interfere with the MuRF1–titin interaction from a 130 000 compound screen based on Alpha Technology. A subset of nine prioritized compounds were synthesized and administrated during conditions of muscle wasting, that is, to C2C12 muscle cells treated with dexamethasone and to mice treated with monocrotaline to induce cardiac cachexia. Results; The nine selected compounds inhibited MuRF1–titin complexation with IC50 values <25 μM, of which three were found to also inhibit MuRF1 E3 ligase activity, with one further showing low toxicity on cultured myotubes. This last compound, EMBL chemical core ID#704946, also prevented atrophy in myotubes induced by dexamethasone and attenuated fibre atrophy and contractile dysfunction in mice during cardiac cachexia. Proteomic and western blot analyses showed that stress pathways were attenuated by ID#704946 treatment, including down‐regulation of MuRF1 and normalization of proteins associated with apoptosis (BAX) and protein synthesis (elF2B‐delta). Furthermore, actin ubiquitinylation and proteasome activity was attenuated. Conclusions; We identified a novel compound directed to MuRF1's central myofibrillar protein recognition domain. This compound attenuated in vivo muscle wasting and contractile dysfunction in cardiac cachexia by protecting de novo protein synthesis and by down‐regulating apoptosis and ubiquitin‐proteasome‐dependent proteolysis

Correction to: Use of MFM-20 to monitor SMA types 1 and 2 patients treated with nusinersen.

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The terrestrialization process: A palaeobotanical and palynological perspective (2)

International audienc

Lymphocyte activation marker analysis in patients with sight threatening uveitis

info:eu-repo/semantics/nonPublishe

Développement de mini-formes de gaufres imprimées en 3D contenant de l'hydrocortisone pour la médecine personnalisée des enfants

peer reviewedHydrocortisone is mainly used in the substitution treatment of adrenal insufficiency which results in a dysregulation of cortisol. Compounding of hydrocortisone capsules remains the only low-dose oral treatment suitable for the pediatric population. However, capsules often show non-compliance in mass and content uniformity. Three-dimensional printing offers the prospect of practising personalized medicine for vulnerable patients like children. The goal of this work is to develop low-dose solid oral forms containing hydrocortisone by hot-melt extrusion coupled with fused deposition modeling for the pediatric population. Formulation, design and processes temperatures were optimized to produce printed forms with the desired characteristics. Red mini-waffle shapes containing drug loads of 2, 5 and 8 mg were successfully printed. This new 3D design allow to release more than 80% of the drug in 45 minutes indicating a conventional release like the one obtained with capsules. Mass and content uniformity, hardness and friability tests complied with European Pharmacopeia specifications, despite the considerable challenge of the small dimensions of the forms. This study demonstrates that FDM can be used to produce innovative pediatric-friendly printed shapes of an advanced pharmaceutical quality to practice personalize medicine.Development of new oral formulations produced by hot-melt extrusion technique coupled with fused deposition modeling 3D printin

Les formes de gaufres imprimées en 3D pourraient-elles remplacer les gélules d'hydrocortisone préparées dans les pharmacies d'officine pour la médecine personnalisée des enfants ?

Development of new oral formulations produced by the hot melt extrusion technique coupled with 3D printing in order to increase the solubility of active molecules like BCSI

L'avenir de l'impression 3D pour la médecine personnalisée : Les formes de gaufres imprimées en 3D pourraient-elles remplacer la préparation magistrale des gélules d'hydrocortisone ?

INTRODUCTION Hydrocortisone (HCT) is mainly used in the substitution treatment of adrenal insufficiency (AS) which results in a disruption of the production of cortisol. The magistral preparation of HCT capsules in community pharmacies remains the only oral treatment adapted to the paediatric population due to the low doses administered. However, these preparations show non-compliance in mass and content uniformity. Three-dimensional (3D) printing appears as a potential solution because of the possibility to practise personalized medicine. The goal of this work is therefore to develop low-dose solid oral forms, of advanced pharmaceutical quality, containing HCT by HME and Fused Deposition Modeling (FDM) for the treatment of children against AS. MATERIALS AND METHODS The formulation contained HCT, Affinisol® 15LV (hydroxypropylmethylcellulose), Kollidon® VA 64 (vinylpyrrolidone-vinyl acetate), tri(ethyl)citrate and red iron oxide. HME was performed at 140°C and 50 rpm. Waffle shapes with diameters of 4.5, 6.0, 7.25 mm, heights of 2.0, 2.8 and 3.2 mm and drug loads of 2, 5 and 8 mg were printed at 155°C (Figure 1). The monograph uncoated tablets in European Pharmacopoeia 11th edition was used to characterize the printed forms. Dissolution studies were performed in 500 mL of HCl 0.1 M during 2 h. The HCT content was evaluated by a validated HPLC method. Friability test was performed at 25 rpm for 4 min. RESULTS AND DISCUSSION More than 80% of HCT was released in 45 min from waffle shapes containing 2, 5 and 8 mg of HCT, indicating a conventional drug release. The content uniformity of all 3D printed waffle shapes has shown encouraging results with maximum acceptance values lower than the requested value of 15.0. The differences in total weight before and after the test were lower than 1.0% for all 3D printed waffle shapes, which complied with the specifications. CONCLUSION Red 3D printed waffle shapes containing low-dose HCT for pediatric use were successfully developed by HME coupled to FDM. Content uniformity, drug dissolution and friability complied with the specifications, despite the considerable challenge of their small dimensions. 3D-printed solid oral forms could offer patients a higher pharmaceutical quality than capsules containing low-dose corticosteroids. Solid oral forms containing different dosages of API can be printed with the same filament, offering the prospect of practicing personalized medicine.Development of new oral formulations produced by the hot melt extrusion technique coupled with 3D printing in order to increase the solubility of active molecules like BCSI

Late-Onset Nephropathic Cystinosis: Clinical Presentation, Outcome, and Genotyping

Background and objectives: Cystinosis is an autosomal recessive disease characterized by the intralysosomal accumulation of cystine, as a result of a defect in cystine transport across the lysosomal membrane. Three clinical forms have been described on the basis of severity of symptoms and age of onset: infantile cystinosis, characterized by renal proximal tubulopathy and progression to end-stage renal disease before 12 yr of age; juvenile form, with a markedly slower rate of progression; and adult form, with only ocular abnormalities

Reduced cardiolipin content decreases respiratory chain capacities and increases ATP synthesis yield in the human HepaRG cells

International audienceCardiolipin (CL) is a unique mitochondrial phospholipid potentially affecting many aspects of mitochondrial function/processes, i.e. energy production through oxidative phosphorylation. Most data focusing on implication of CL content and mitochondrial bioenergetics were performed in yeast or in cellular models of Barth syndrome. Previous work reported that increase in CL content leads to decrease in liver mitochondrial ATP synthesis yield. Therefore the aim of this study was to determine the effects of moderate decrease in CL content on mitochondrial bioenergetics in human hepatocytes. For this purpose, we generated a cardiolipin synthase knockdown (shCLS) in HepaRG hepatoma cells showing bioenergetics features similar to primary human hepatocytes. shCLS cells exhibited a 55% reduction in CLS gene and a 40% decrease in protein expression resulting in a 45% lower content in CL compared to control (shCTL) cells. Oxygen consumption was significantly reduced in shCLS cells compared to shCTL regardless of substrate used and energy state analyzed. Mitochondrial low molecular weight supercomplexes content was higher in shCLS cells (+ 60%) compared to shCTL. Significant fragmentation of the mitochondrial network was observed in shCLS cells compared to shCTL cells. Surprisingly, mitochondrial ATP synthesis was unchanged in shCLS compared to shCTL cells but exhibited a higher ATP:O ratio (+ 46%) in shCLS cells. Our results suggest that lowered respiratory chain activity induced by moderate reduction in CL content may be due to both destabilization of supercomplexes and mitochondrial network fragmentation. In addition, CL content may regulate mitochondrial ATP synthesis yield