L-Carnitine Functionalization to Increase Skeletal Muscle Tropism of PLGA Nanoparticles

Muscular dystrophies are a group of rare genetic pathologies, encompassing a variety of clinical phenotypes and mechanisms of disease. Several compounds have been proposed to treat compromised muscles, but it is known that pharmacokinetics and pharmacodynamics problems could occur. To solve these issues, it has been suggested that nanocarriers could be used to allow controlled and targeted drug release. Therefore, the aim of this study was to prepare actively targeted poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) for the treatment of muscular pathologies. By taking advantage of the high affinity for carnitine of skeletal muscle cells due to the expression of Na⁺-coupled carnitine transporter (OCTN), NPs have been actively targeted via association to an amphiphilic derivative of L-carnitine. Furthermore, pentamidine, an old drug repurposed for its positive effects on myotonic dystrophy type I, was incorporated into NPs. We obtained monodispersed targeted NPs, with a mean diameter of about 100 nm and a negative zeta potential. To assess the targeting ability of the NPs, cell uptake studies were performed on C2C12 myoblasts and myotubes using confocal and transmission electron microscopy. The results showed an increased uptake of carnitine-functionalized NPs compared to nontargeted carriers in myotubes, which was probably due to the interaction with OCTN receptors occurring in large amounts in these differentiated muscle cells.     

Cerebral Folate Metabolism in Post-Mortem Alzheimer’s Disease Tissues: A Small Cohort Study

We investigated the cerebral folate system in post-mortem brains and matched cerebrospinal fluid (CSF) samples from subjects with definite Alzheimer’s disease (AD) (n = 21) and neuropathologically normal brains (n = 21) using immunohistochemistry, Western blot and dot blot. In AD the CSF showed a significant decrease in 10-formyl tetrahydrofolate dehydrogenase (FDH), a critical folate binding protein and enzyme in the CSF, as well as in the main folate transporter, folate receptor alpha (FRα) and folate. In tissue, we found a switch in the pathway of folate supply to the cerebral cortex in AD compared to neurologically normal brains. FRα switched from entry through FDH-positive astrocytes in normal, to entry through glial fibrillary acidic protein (GFAP)-positive astrocytes in the AD cortex. Moreover, this switch correlated with an apparent change in metabolic direction to hypermethylation of neurons in AD. Our data suggest that the reduction in FDH in CSF prohibits FRα-folate entry via FDH-positive astrocytes and promotes entry through the GFAP pathway directly to neurons for hypermethylation. This data may explain some of the cognitive decline not attributable to the loss of neurons alone and presents a target for potential treatment.     

Limited Accuracy of Pan-Trk Immunohistochemistry Screening for NTRK Rearrangements in Follicular-Derived Thyroid Carcinoma

Patients with advanced thyroid cancer harboring NTRK rearrangements can be treated with highly effective selective inhibitors. Immunohistochemistry (IHC) analysis, to detect Trk protein expression, represents an appealing screening strategy for NTRK rearrangements, but its efficacy has been poorly explored in thyroid cancer. The aim of this study is to investigate the diagnostic utility of Trk IHC in the identification of NTRK rearrangements. A series of 26 follicular-derived thyroid tumors, positive for NTRK rearrangements, and 28 NTRK fusion-negative controls were retrospectively analyzed by IHC using the pan-Trk monoclonal antibody (clone {"type":"entrez-protein","attrs":{"text":"EPR17341","term_id":"523383444","term_text":"EPR17341"}}EPR17341) on the Ventana system. Area under the curve (AUC), sensitivity and specificity were calculated by ROC analysis. Trk expression was detected in 25 samples, including 22 out of the 26 NTRK-rearranged (84.6%) and three out of 28 NTRK-negative samples (10.7%). Four out of twenty-six NTRK-rearranged thyroid tumors were negative for Trk expression (15.4%), all carrying the ETV6/NTRK3 fusion. The AUC, sensitivity and specificity were 0.87, 0.85 and 0.89, respectively. A screening based on IHC analysis showed limited sensitivity and specificity in the identification of NTRK-rearranged tumors. Since falsely negative results could preclude the administration of effective targeted drugs, alternative detection strategies should be considered for thyroid cancer.     

Cholesterol photosensitised oxidation of horse meat slices stored under different packaging films

The effect of the type of packaging film (transparent vs. light-protecting red film) was evaluated on the formation of cholesterol oxidation products (COPs) in refrigerated horse meat slices stored in retail conditions under light exposure for 8 h. In meat wrapped with a transparent film, COPs increased from 233 (control) to 317 μg/g of fat, whereas the red film delayed cholesterol oxidation and offered protection against COPs formation, since COPs decreased from 173 (control) to 139 μg/g of fat after 8 h of light exposure. In addition, light opened the epoxy ring and led to the formation of triol, which was actually absent at T_0. A proper packaging film may represent a useful strategy to retard oxidative degradation in a light-sensitive, high pigment- and fat-containing food, such as horse meat.     

Pigment composition and color parameters of commercial Spanish red wine samples: linkage to quality perception

Anthocyanins play a crucial role in wine color. Thus, their analysis results essential for evaluating their contribution to perceived color in wines, attribute directly linked to quality judgements. In this context, this study was aimed at: (1) defining the color space of a relative large number of commercial oaked Spanish red wines and (2) establishing a linkage between wine coloring, anthocyanic composition, and quality perception of this set of wines. Therefore, CIELab parameters (a ₁₀*, b ₁₀*, L ₁₀*), monomeric anthocyanin contents analyzed by HPLC with ultraviolet and mass detection, and polymeric pigment content estimated by a protein precipitation assay combined with bisulfite bleaching were evaluated in a total of 58 wine samples. Principal component analysis was performed on color variables, and their correlations were discussed. The results have evidenced the important role played by the pyranoanthocyanic compounds since they seem to be the main contributors to the red color of these aged wines, and they have been demonstrated not to increase their yellow nuances. On the other hand, results highlight that, regardless of the time of aging, wines presenting more red (higher values for a ₁₀* and for low molecular weighted anthocyanins) and lower yellow nuances (lower values for b ₁₀* and T) as well as a darker color (lower in L ₁₀* and higher in CI) are in general evaluated with higher quality scores by wine experts.     

Genomic Analysis Made It Possible to Identify Gene-Driver Alterations Covering the Time Window between Diagnosis of Neuroblastoma 4S and the Progression to Stage 4

Neuroblastoma (NB) is a tumor of the developing sympathetic nervous system. Despite recent advances in understanding the complexity of NB, the mechanisms that determine its regression or progression are still largely unknown. Stage 4S NB is characterized by a favorable course of disease and often by spontaneous regression, while progression to true stage 4 is a very rare event. Here, we focused on genomic analysis of an NB case that progressed from stage 4S to stage 4 with a very poor outcome. Array-comparative genomic hybridization (a-CGH) on tumor-tissue DNA, and whole-exome sequencing (WES) on exosomes DNA derived from plasma collected at the onset and at the tumor progression, pointed out relevant genetic changes that can explain this clinical worsening. The combination of a-CGH and WES data allowed for the identification iof somatic copy number aberrations and single-nucleotide variants in genes known to be responsible for aggressive NB. KLRB1, MAPK3 and FANCA genes, which were lost at the time of progression, were studied for their possible role in this event by analyzing in silico the impact of their expression on the outcome of 786 NB patients.     

A Lagrange duality characterisation for stability under arbitrary switching in switched positive linear systems

The present communication is concerned with uniform exponential stability, under arbitrary switching, in discrete-time switched positive linear systems. Lagrange duality is used in order to obtain a new characterisation for uniform exponential stability which is in terms of sets of inequalities involving each of the matrices that represent the modes of the system. These sets of inequalities are shown to generalise the classical linear Lyapunov inequality that characterises, in positive matrices, the property of being Schur. Each solution to these sets of inequalities is shown to provide a representation, in terms of a number of linear functionals, for a common Lyapunov function for the switched positive linear system. A result is further presented which conveys to, a conservative upper bound on the minimum required number of linear functionals (in the above mentioned representation), and also to a method for computing them. Our proof for the aforementioned characterisation is based on another (equivalent) characterisation, in terms of the solvability of a dynamic programming equation associated to the switched positive linear system, which is also reported in the paper. In particular, it is shown that the associated dynamic programming equation has at most one solution. And this solution is shown to be convex, monotonic, positively homogeneous, and it yields a common Lyapunov function for the switched positive linear system.     

Reduction of the Nitrate Content in Baby Spinach Leaves by Vacuum Impregnation with Sucrose

Vacuum impregnation (VI) has been widely utilized as a pre-treatment method prior to, e.g., the minimal processing, freezing, or drying of foods. In many cases, VI has been used to focus on the enrichment of fruits and vegetables with probiotics or micronutrients, texture enhancement, the modification of the sensory attributes, and extension of shelf life by pH reduction. However, little attention has been paid to the exploration of the metabolic consequences of VI that could lead to changes in the quality characteristics of plant tissues. Since nitrate has long been discussed as a compound that is harmful to human health, the aim of this investigation was to reduce the nitrate content in spinach leaves by feeding sucrose into the tissue using VI. The leaves, either non-treated or treated with VI, were stored under saturated humidity conditions and in darkness at 8 °C, for up to 72 h. VI-treated leaves showed a remarkable reduction in nitrate content as compared to the non-treated samples. Upon storage, sucrose was reduced, indicating that this sugar had been respired and had induced metabolization of the stored nitrate. The nitrite content of the treated leaves was unaffected, proving that this toxic compound was not accumulated in the baby spinach leaves upon external sucrose feeding.