Lysosomes serve as the degradation hubs for macroautophagic/autophagic and endocytic components, thus maintaining cellular homeostasis essential for neuronal survival and function. LAMP1 (lysosomal ...associated membrane protein 1) and LAMP2 are distributed among autophagic and endolysosomal organelles. Despite widespread distribution, LAMP1 is routinely used as a lysosome marker and LAMP1-positive organelles are often referred to as lysosomal compartments. By applying immuno-electron microscopy (iTEM) and confocal imaging combined with Airyscan microscopy, we expand on the limited literature to provide a comprehensive and quantitative analysis of LAMP1 distribution in various autophagic and endolysosomal organelles in neurons. Our study demonstrates that a significant portion of LAMP1-labeled organelles lack major lysosomal hydrolases. BSA-gold pulse-chase assay further shows heterogeneous degradative capacities of LAMP1-labled organelles. In addition, LAMP1 intensity is not a sensitive readout to assess lysosomal deficits in familial amyotrophic lateral sclerosis-linked motor neurons in vivo. Our study thus calls for caution when interpreting LAMP1-labeled organelles in the nervous system where LAMP1 intensity, trafficking, and distribution do not necessarily represent degradative lysosomes or autolysosomes under physiological and pathological conditions.
Abbreviations: ALS: amyotrophic lateral sclerosis; BSA: bovine serum albumin; DRG: dorsal root ganglion; IGF2R/CI-M6PR: insulin like growth factor 2 receptor; iTEM: immuno-transmission electron microscopy; LAMP1/2: lysosomal associated membrane protein 1/2; P80: postnatal day 80; sMNs: spinal motor neurons
Cet article présente les résultats de la recherche-action du programme Histoires de famille ayant été offert à 9 familles issues de milieux défavorisés pendant 12 semaines dans divers lieux ...communautaires (p. ex., bibliothèque municipale, musée, maison de la famille, etc.). Ce programme a pour but d’outiller les parents à faire des activités de littératie afin de favoriser le développement langagier des enfants. L’objectif de cette étude qualitative est d’explorer les pratiques en littératie de mères issues de milieux défavorisés participant au projet communautaire Histoires de famille, de même que l’évolution de leurs pratiques au cours du projet. Les résultats montrent que les mères ont apprécié l’approche utilisée dans le programme, ont intégré des activités de littératie dans leur quotidien et ont constaté plusieurs apprentissages qu’elles-mêmes et leurs enfants ont réalisés. Des changements ont également été observés dans la manière dont les mères offrent du soutien à leurs enfants dans la réalisation de tâches de littératie. Enfin, cette recherche révèle l’importance d’ouvrir les portes des organismes communautaires aux familles vulnérables en les invitant à vivre des activités de littératie pour favoriser la préparation scolaire.
Paclitaxel and nanoparticle albumin-bound paclitaxel are known to cause adverse events of eye disorders, such as cystoid macular edema. However, at present, the risk factors remain unclear. ...Therefore, risk factors for eye disorders caused by paclitaxel and nanoparticle albumin-bound paclitaxel were studied. This retrospective study targeted patients who were newly administered paclitaxel or nanoparticle albumin-bound paclitaxel at Kyoto Okamoto Memorial Hospital between April 1, 2012, and March 31, 2017. Eye disorder occurrence was defined as an event in which the pharmacist confirmed the symptoms in a patient interview and the ophthalmologist diagnosed the disorder. To analyze the risk factors, logistic regression analysis using 41 factors was performed. Of 128 subjects, 13 (10.2%) had eye disorders with symptom degrees of Grades 1 and 2. The symptoms were conjunctivitis or subconjunctival hemorrhage (3.1%), visual acuity reduction (2.3%), blurred vision and eye pain (1.6% each), eye mucus, blepharitis, stye, watering eyes, photopsia, and muscae volitantes (0.8% each). In eight patients, the conditions patients improved with spontaneously or with medication use; no improvements were observed the cases of visual acuity reduction, blurred vision, or muscae volitantes. Multivariate logistic regression analysis revealed that a cumulative dose of >-819 mg/m2 (odds ratio: 5.34, 95% confidence interval: 1.32-21.60, p=0.019) and baseline alkaline phosphatase >-256U/L (odds ratio: 3.74, 95% confidence interval: 1.02-13.70, p=0.046) were significant risk factors associated with eye disorders. In conclusion, it was determined that paclitaxel- and nanoparticle albumin-bound paclitaxel-related eye disorders might be influenced by cumulative dose and baseline alkaline phosphatase.
Un rapide rappel des traits définitoires du genre metal nous permet de poser l’existence, dans cette musique, de trois catégories distinctes d’albums-concept : anthologie, geste, épopée. Leur examen ...nous amène à nous demander si ce format particulier d’album a pour vocation de dérégler, par le truchement intermédial, ce qui avait forme et légitimité, ou, au contraire, de mettre de l’ordre dans le chaos d’une musique extrême.
The objective of this work was to investigate the mechanism of release of active pharmaceutical ingredients (APIs) both small molecules (ketoprofen, indomethacin, coumarin-6) and macromolecules ...(human serum albumin, ovalbumin), from PLGA (50:50) nanoparticulates (400–700
nm), having drug loadings less than 10%. The nanoparticulates were prepared by emulsification/solvent evaporation methods and release determined in phosphate buffer pH 7.4 at 37
°C. The release profiles exhibited an initial burst release phase, a slower lag phase and a second increased release rate phase. The profiles were consistent with a model in which the first phase of the release reflects diffusion controlled dissolution of drug accessible to the solid/dissolution medium interface and the second phase reflects release of drug entrapped in the polymer, the release of which was dependent on the bulk degradation of the polymer. The burst phase tended to increase with increase in API loading and solubility. The polymer erosion related parameters also indicated that increased drug loading accelerated this phase of API release. Small acidic hydrophobic actives such as ketoprofen and indomethacin had a much greater effect on these parameters than the larger hydrophilic more neutral proteins, HSA and ovalbumin.
This work provides a new understanding of critical process parameters involved in the production of inhalation aerosol particles by ultrasonic spray freeze drying to enable precise control over ...particle size and aerodynamic properties. A series of highly porous mannitol, lysozyme, and bovine serum albumin (BSA) particles were produced, varying only the solute concentration in the liquid feed, cs, from 1 to 5wt%. The particle sizes of mannitol, BSA, and lysozyme powders were independent of solute concentration, and depend only on the drop size produced by atomization. Both mannitol and lysozyme formulations showed a linear relationship between the computed Fine Particle Fraction (FPF) and the square root of cs, which is proportional to the particle density, ρ, given a constant particle size dg. The FPF decreased with increasing cs from 57.0% to 16.6% for mannitol and 44.5% to 17.2% for lysozyme. Due to cohesion, the BSA powder FPF measured by cascade impaction was less than 10% and independent of cs. Ultrasonic spray freeze drying enables separate control over particle size, dg, and aerodynamic size, da which has allowed us to make the first experimental demonstration of the widely accepted rule da=dg(ρ/ρo)1/2 with particles of constant dg, but variable density, ρ (ρo is unit density).
To identify determinants of early progressive renal decline in type 2 diabetes a range of markers was studied in 1032 patients enrolled into the 2nd Joslin Kidney Study. eGFR slopes estimated from ...serial measurements of serum creatinine during 5-12 years of follow-up were used to define early renal decline. At enrollment, all patients had normal eGFR, 58% had normoalbuminuria and 42% had albuminuria. Early renal decline developed in 6% and in 18% patients, respectively. As determinants, we examined baseline values of clinical characteristics, circulating markers: TNFR1, KIM-1, and FGF23, and urinary markers: albumin, KIM-1, NGAL, MCP-1, EGF (all normalized to urinary creatinine) and the ratio of EGF to MCP-1. In univariate analysis, all plasma and urinary markers were significantly associated with risk of early renal decline. When analyzed together, systolic blood pressure, TNFR1, KIM-1, the albumin to creatinine ratio, and the EGF/MCP-1 ratio remained significant with the latter having the strongest effect. Integration of these markers into a multi-marker prognostic test resulted in a significant improvement of discriminatory performance of risk prediction of early renal decline, compared with the albumin to creatinine ratio and systolic blood pressure alone. However, the positive predictive value was only 50% in albuminuric patients. Thus, markers in plasma and urine indicate that the early progressive renal decline in Type 2 diabetes has multiple determinants with strong evidence for involvement of tubular damage. However, new, more informative markers are needed to develop a better prognostic test for such decline that can be used in a clinical setting.
Mammalian cells have been shown to internalize oligonucleotide-functionalized gold nanoparticles (DNA-Au NPs or siRNA-Au NPs) without the aid of auxiliary transfection agents and use them to initiate ...an antisense or RNAi response. Previous studies have shown that the dense monolayer of oligonucleotides on the nanoparticle leads to the adsorption of serum proteins and facilitates cellular uptake. Here, we show that serum proteins generally act to inhibit cellular uptake of DNA-Au NPs. We identify the pathway for DNA-Au NP entry in HeLa cells. Biochemical analyses indicate that DNA-Au NPs are taken up by a process involving receptor-mediated endocytosis. Evidence shows that DNA-Au NP entry is primarily mediated by scavenger receptors, a class of pattern-recognition receptors. This uptake mechanism appears to be conserved across species, as blocking the same receptors in mouse cells also disrupted DNA-Au NP entry. Polyvalent nanoparticles functionalized with siRNA are shown to enter through the same pathway. Thus, scavenger receptors are required for cellular uptake of polyvalent oligonucleotide functionalized nanoparticles.
O-Phosphoethanolamine (PEA) is an endogenous substance that is attracting interest as a biomarker for depression, and thus there is a need to develop a simple analytical method that specifically ...measures PEA. Therefore, this study aimed to develop a simple and specific enzyme-linked immunosorbent assay (ELISA) for PEA.
Anti-PEA antibody was obtained by immunizing mice with an antigen conjugated with mercaptosuccinyl bovine serum albumin using m-maleimidobenzoyl-N-hydroxysulfosuccinimide ester (MBS). In this assay, the PEA to be quantified is chemically modified by benzoyl chloride that is allowed to compete with a PEA-MBS-HRP conjugate for binding to a limited amount of an anti-PEA antibody, which was used to coat the wells of a microtiter plate. This ELISA shows a linear range of detection of 0.11–27 μM, and a limit of quantification of 0.144 μM. The anti-PEA antibody showed high affinity for benzoyl PEA. No detectable cross-reactivity was found with benzoyl 2-aminoethanol, O-phospho-l-tyrosine or benzoyl sphingosine-1-phosphate. The values of plasma PEA levels measured by this ELISA were comparable to those measured by HPLC, and a strong correlation was observed between the values determined by the two methods. The developed ELISA should provide a valuable new tool for the quantification of PEA in human plasma.
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•We developed an ELISA for PEA, a biomarker of depression.•This is the first ELISA that exhibits specificity for PEA.•The limit of quantification is 0.144 μM for PEA.•This method is useful for the quantification of PEA in human plasma.
È ormai noto come la produzione digitale abbia quasi del tutto soppiantato le tradizionali pratiche del disegno nel processo architettonico. Se da un lato l’uso della rappresentazione digitale ha ...interrotto la produzione grafica tradizionale, dall’altro essa rende possibile una sempre più efficace e immersiva disseminazione e divulgazione di progetti architettonici antichi non sufficientemente conosciuti, se non inediti o, ancora, immaginati. In questo senso, il presente contributo vuole indagare e sviluppare la digitalizzazione di uno dei più importanti progetti in ambito costiero posto in essere da parte del Ministero dei Lavori Pubblici e concretizzato mediante la redazione dell’Album dei fari illustrato dalle notizie intorno ai loro caratteri e posizione, al fine di rendere fruibili tutti quei progetti in tutto o in parte realizzati per la crescita delle nostre coste. Il progetto proposto nel 1873, all’indomani dell’Unità d’Italia, prevede la progettazione e la realizzazione di quattrocentosessantadue segnalamenti marittimi per gli ottomila chilometri di costa italiana. La digitalizzazione dell’Album dei fari, ovvero un caposaldo per la storia della costruzione di queste architetture costiere, si configura dunque come l’azione fondamentale per intraprendere un percorso di conoscenza e comparazione di tipo digitale delle strutture costiere passate e presenti, secondo i dettami dell’interdisciplinarietà Albisinni, De Carlo 2011.
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