Rescuing a Troubled Tolcapone with PEGylated PLGA Nanoparticles: Design, Characterization, and Hepatotoxicity Evaluation.

Tolcapone is an orally active catechol-O-methyltransferase (COMT) inhibitor used as adjuvant therapy in Parkinson's disease. However, it has a highly hepatotoxic profile, as recognized by the U.S. Food and Drug Administration. As a possible solution, nanoscience brought us several tools in the development of new functional nanomaterials with tunable physicochemical properties, which can be part of a solution to solve several drawbacks, including drug's short half-life and toxicity. This work aims to use PEGylated poly(lactic-co-glycolic acid) (PLGA) nanoparticles as a stable carrier with lower hydrodynamic size and polydispersity to encapsulate tolcapone in order to overcome its therapeutic drawbacks. Using the nanoprecipitation method, tolcapone-loaded nanoparticles with a DLC% of 5.7% were obtained (EE% of 47.0%) and subjected to a lyophilization optimization process to obtain a final shelf-stable formulation. Six different cryoprotectants in concentrations up to 10% (w/v) were tested. A formulation of PLGA nanoparticles with 3% hydroxypropyl-ß-cyclodextrin (HPßCD) as a cryoprotectant (PLGA-HP@Tolc), presenting sub-200 nm sizes and low polydispersity (PdI < 0.200) was selected. Cytotoxicity assays, namely, MTT and SRB, were used to study the metabolic activity and cell density of tolcapone and PLGA-HP@Tolc-treated cells. In both assays, a hepatocarcinoma cell line (HepG2) growing in glucose or glucose-free media (galactose-supplemented medium) was used. The results demonstrated that the treatment with the PLGA-HP@Tolc formulation led to a decrease in cytotoxicity in comparison to free tolcapone-treated cells in both media tested. Moreover, the elected formulation also counteracted ATP-depletion and excessive ROS production induced by tolcapone. The results suggest that HPßCD might have a dual function in the formulation: cryoprotectant and anticytotoxic agent, protecting cells from tolcapone-induced damage. Using an in vitro COMT inhibition assay, the PLGA-HP@Tolc formulation demonstrated to inhibit COMT as efficiently as free tolcapone. Overall, the results suggest that tolcapone-loaded PLGA NPs could be an interesting alternative to free tolcapone, demonstrating the same in vitro efficacy in inhibiting COMT but with a safer cytotoxic profile.

Management and outcomes of bronchiolitis in Italy and Latin America: a multi-center, prospective, observational study.

We aimed to describe differences in the epidemiology, management, and outcomes existing between centers located in countries which differ by geographical location and economic status during to post-pandemic bronchiolitis seasons.  This was a prospective observational cohort study performed in two academic centers in Latin America (LA) and three in Italy. All consecutive children with a clinical diagnosis of bronchiolitis were included, following the same data collection form.  Nine hundred forty-three patients have been enrolled: 275 from the two Latin American Centers (San Jose, 215; Buenos Aires, 60), and 668 from Italy (Rome, 178; Milano, 163; Bologna, 251; Catania, 76). Children in LA had more frequently comorbidities, and only rarely received palivizumab. A higher number of patients in LA had been hospitalized in a ward (64% versus 23.9%, p < 0.001) or in a PICU (16% versus 6.2%, p < 0.001), and children in LA required overall more often respiratory support, from low flow oxygen to invasive mechanical ventilation, except for CPAP which was more used in Italy. There was no significant difference in prescription rates for antibiotics, but a significantly higher number of patients treated with systemic steroids in Italy. CONCLUSIONS: We found significant differences in the care for children with bronchiolitis in Italy and LA. Reasons behind such differences are unclear and would require further investigations to optimize and homogenize practice all over the world. WHAT IS KNOWN: • Bronchiolitis is among the commest cause of morbidity and mortality in infants all over the world. WHAT IS NEW: • There are significant differences on how clinicians care for bronchiolitis in different centers and continents. Differences in care can be principally due to different local practices than differences in patients severity/presentations. • Understanding these differences should be a priority to optime and standardize bronchiolitis care globally.

Ataxia cerebelosa aguda y COVID-19: presentación de un caso clínico / Ataxia cerebelosa aguda y COVID-19: presentación de un caso clínico

COVID-19 es una enfermedad viral principalmente respiratoria y/o gastrointestinal. Las manifestaciones neurológicas tienen una frecuencia variable en pediatría. Presentamos un varón de 10 años de edad, previamente sano, que presentó una ataxia cerebelosa durante un cuadro agudo de COVID-19. El SARS-CoV-2 fue detectado por hisopado nasofaríngeo por antígeno y RPC. El LCR fue normal y el cultivo bacteriológico y estudio viral fueron negativos. La TC y RM encefálica fueron normales. No requirió tratamiento específico y tuvo una evolución favorable, con resolución completa de los síntomas neurológicos al mes. Debe considerarse la infección por SARS-CoV-2 como un diagnóstico diferencial entre las causas de ataxia cerebelosa aguda, según la situación epidemiológica.

COVID-19 is a disease that mainly produces respiratory and/or gastrointestinal symptoms. Neurological manifestations occur with a variable frequency in children. We present a previously healthy 10-year-old boy who presented acute cerebellar ataxia during an acute COVID-19. SARS-CoV-2 was detected in a nasopharyngeal sample by antigen and PCR. The CSF was normal, the bacteriological culture and the viral PCR were negative. CT of the brain and gadolinium MRI of the brain were normal. He did not require specific treatment and had a favorable evolution, with complete resolution of neurological symptoms at one month. SARS-CoV-2 infection should be considered as a differential diagnosis between the causes of acute cerebellar ataxia, according to the epidemiological situation.

Modulating Cytotoxicity with Lego-like Chemistry: Upgrading Mitochondriotropic Antioxidants with Prototypical Cationic Carrier Bricks.

Although the lipophilic triphenylphosphonium (TPP+) cation is widely used to target antioxidants to mitochondria, TPP+-based derivatives have shown cytotoxicity in several biological in vitro models. We confirmed that Mito.TPP is cytotoxic to both human neuronal (SH-SY5Y) and hepatic (HepG2) cells, decreasing intracellular adenosine triphosphate (ATP) levels, leading to mitochondrial membrane depolarization and reduced mitochondrial mass after 24 h. We surpassed this concern using nitrogen-derived cationic carriers (Mito.PICO, Mito.ISOQ, and Mito.IMIDZ). As opposed to Mito.TPP, these novel compounds were not cytotoxic to SH-SY5Y and HepG2 cells up to 50 µM and after 24 h of incubation. All of the cationic derivatives accumulated inside the mitochondrial matrix and acted as neuroprotective agents against iron(III), hydrogen peroxide, and tert-butyl hydroperoxide insults. The overall data showed that nitrogen-based cationic carriers can modulate the biological performance of mitochondria-directed antioxidants and are an alternative to the TPP cation.

Fiscalin Derivatives as Potential Neuroprotective Agents.

Neurodegenerative diseases (ND) share common molecular/cellular mechanisms that contribute to their progression and pathogenesis. In this sense, we are here proposing new neuroprotection strategies by using marine-derived compounds as fiscalins. This work aims to evaluate the protective effects of fiscalin derivatives towards 1-methyl-4-phenylpyridinium (MPP+)- and iron (III)-induced cytotoxicity in differentiated SH-SY5Y cells, an in vitro disease model to study ND; and on P-glycoprotein (P-gp) transport activity, an efflux pump of drugs and neurotoxins. SH-SY5Y cells were simultaneously exposed to MPP+ or iron (III), and noncytotoxic concentrations of 18 fiscalin derivatives (0-25 µM), being the cytotoxic effect of both MPP+ and iron (III) evaluated 24 and 48 h after exposure. Fiscalins 1a and 1b showed a significant protective effect against MPP+-induced cytotoxicity and fiscalins 1b, 2b, 4 and 5 showed a protective effect against iron (III)-induced cytotoxicity. Fiscalins 4 and 5 caused a significant P-gp inhibition, while fiscalins 1c, 2a, 2b, 6 and 11 caused a modest increase in P-gp transport activity, thus suggesting a promising source of new P-gp inhibitors and activators, respectively. The obtained results highlight fiscalins with promising neuroprotective effects and with relevance for the synthesis of new derivatives for the treatment/prevention of ND.

Brain drug delivery and neurodegenerative diseases: Polymeric PLGA-based nanoparticles as a forefront platform.

The discovery of effective drugs for the treatment of neurodegenerative disorders (NDs) is a deadlock. Due to their complex etiology and high heterogeneity, progresses in the development of novel NDs therapies have been slow, raising social/economic and medical concerns. Nanotechnology and nanomedicine evolved exponentially in recent years and presented a panoply of tools projected to improve diagnosis and treatment. Drug-loaded nanosystems, particularly nanoparticles (NPs), were successfully used to address numerous drug glitches, such as efficacy, bioavailability and safety. Polymeric nanoparticles (PNPs), mainly based on polylactic-co-glycolic acid (PLGA), have been already validated and approved for the treatment of cancer, neurologic dysfunctions and hormonal-related diseases. Despite promising no PNPs-based therapy for neurodegenerative disorders is available up to date. To stimulate the research in the area the studies performed so far with polylactic-co-glycolic acid (PLGA) nanoparticles as well as the techniques aimed to improve PNPs BBB permeability and drug targeting were revised. Bearing in mind NDs pharmacological therapy landscape huge efforts must be done in finding new therapeutic solutions along with the translation of the most promising results to the clinic, which hopefully will converge in the development of effective drugs in a foreseeable future.

In Vitro Studies on Nasal Formulations of Nanostructured Lipid Carriers (NLC) and Solid Lipid Nanoparticles (SLN).

The nasal route has been used for many years for the local treatment of nasal diseases. More recently, this route has been gaining momentum, due to the possibility of targeting the central nervous system (CNS) from the nasal cavity, avoiding the blood-brain barrier (BBB). In this area, the use of lipid nanoparticles, such as nanostructured lipid carriers (NLC) and solid lipid nanoparticles (SLN), in nasal formulations has shown promising outcomes on a wide array of indications such as brain diseases, including epilepsy, multiple sclerosis, Alzheimer's disease, Parkinson's disease and gliomas. Herein, the state of the art of the most recent literature available on in vitro studies with nasal formulations of lipid nanoparticles is discussed. Specific in vitro cell culture models are needed to assess the cytotoxicity of nasal formulations and to explore the underlying mechanism(s) of drug transport and absorption across the nasal mucosa. In addition, different studies with 3D nasal casts are reported, showing their ability to predict the drug deposition in the nasal cavity and evaluating the factors that interfere in this process, such as nasal cavity area, type of administration device and angle of application, inspiratory flow, presence of mucoadhesive agents, among others. Notwithstanding, they do not preclude the use of confirmatory in vivo studies, a significant impact on the 3R (replacement, reduction and refinement) principle within the scope of animal experiments is expected. The use of 3D nasal casts to test nasal formulations of lipid nanoparticles is still totally unexplored, to the authors best knowledge, thus constituting a wide open field of research.

Mapping Chromone-3-Phenylcarboxamide Pharmacophore: Quid Est Veritas?

Chromone-3-phenylcarboxamides (Crom-1 and Crom-2) were identified as potent, selective, and reversible inhibitors of human monoamine oxidase B (hMAO-B). Since they exhibit some absorption, distribution, metabolism, and excretion (ADME)-toxicity liabilities, new derivatives were synthesized to map the chemical structural features that compose the pharmacophore, a process vital for lead optimization. Structure-activity relationship data, supported by molecular docking studies, provided a rationale for the contribution of the heterocycle's rigidity, the carbonyl group, and the benzopyran heteroatom for hMAO-B inhibitory activity. From the study, N-(3-chlorophenyl)-4H-thiochromone-3-carboxamide (31) (hMAO-B IC50 = 1.52 ± 0.15 nM) emerged as a reversible tight binding inhibitor with an improved pharmacological profile. In in vitro ADME-toxicity studies, compound 31 showed a safe cytotoxicity profile in Caco-2, SH-SY5Y, HUVEC, HEK-293, and MCF-7 cells, did not present cardiotoxic effects, and did not affect P-gp transport activity. Compound 31 also protected SH-SY5Y cells from iron(III)-induced damage. Collectively, these studies highlighted compound 31 as the first-in-class and a suitable candidate for in vivo preclinical investigation.

Coordination Compounds As Multi-Delivery Systems for Osteoporosis.

Osteoporosis therapies leveraging bisphosphonates and mineral components (e.g., magnesium, calcium, and strontium) have been raising attention because of their potential for managing this ever-growing disease. The administration of multicomponent therapeutics (combined therapy) in elderly patients is complex and suffers from low patient adherence. Herein, we report an all-in-one combination of four antiosteoporotic components into a new family of coordination complexes: [M2(H4alen)4(H2O)2]·1.5H2O [where M2+ = Mg2+ (1), (Mg0.535Ca0.465)2+ (2) and (Mg0.505Ca0.450Sr0.045)2+ (3)]. These solid-state complexes were prepared, for the first time, through microwave-assisted synthesis. It is demonstrated that the compounds are capable of releasing their antiosteoporotic components, both in conditions that mimic the path along the gastrointestinal tract and in long periods under physiological conditions (pH ∼7.4). More importantly, when administered in low concentrations, the compounds did not elicit a cytotoxic effect toward liver, kidney, and osteoblast-like cell lines. Besides, it is important to highlight the unique coordination complex with four bone therapeutic components, [(Mg0.505Ca0.450Sr0.045)2(H4alen)4(H2O)2]·1.5H2O (3), which significantly promoted osteoblast metabolic activity up to ca. 1.4-fold versus the control group. These findings bring this type of compounds one-step closer to be considered as an all-in-one and more effective treatment for managing chronic bone diseases, prompting further research on their therapeutic properties.

Fine-Tuning the Biological Profile of Multitarget Mitochondriotropic Antioxidants for Neurodegenerative Diseases.

Neurotransmitter depletion and mitochondrial dysfunction are among the multiple pathological events that lead to neurodegeneration. Following our previous studies related with the development of multitarget mitochondriotropic antioxidants, this study aims to evaluate whether the π-system extension on the chemical scaffolds of AntiOXCIN2 and AntiOXCIN3 affects their bioactivity and safety profiles. After the synthesis of four triphenylphosphonium (TPP+) conjugates (compounds 2-5), we evaluated their antioxidant properties and their effect on neurotransmitter-metabolizing enzymes. All compounds were potent equine butyrylcholinesterase (eqBChE) and moderate electric eel acetylcholinesterase (eeAChE) inhibitors, with catechols 4 and 5 presenting lower IC50 values than AntiOXCIN2 and AntiOXCIN3, respectively. However, differences in the inhibition potency and selectivity of compounds 2-5 towards non-human and human cholinesterases (ChEs) were observed. Co-crystallization studies with compounds 2-5 in complex with human ChEs (hChEs) showed that these compounds exhibit different binging modes to hAChE and hBChE. Unlike AntiOXCINs, compounds 2-5 displayed moderate human monoamine oxidase (hMAO) inhibitory activity. Moreover, compounds 4 and 5 presented higher ORAC-FL indexes and lower oxidation potential values than the corresponding AntiOXCINs. Catechols 4 and 5 exhibited broader safety windows in differentiated neuroblastoma cells than benzodioxole derivatives 2 and 3. Compound 4 is highlighted as a safe mitochondria-targeted antioxidant with dual ChE/MAO inhibitory activity. Overall, this work is a contribution for the development of dual therapeutic agents addressing both mitochondrial oxidative stress and neurotransmitter depletion.

New marine-derived indolymethyl pyrazinoquinazoline alkaloids with promising antimicrobial profiles.

Due to the emergence of multidrug-resistant pathogenic microorganisms, the search for novel antimicrobials is urgent. Inspired by marine alkaloids, a series of indolomethyl pyrazino [1,2-b]quinazoline-3,6-diones was prepared using a one-pot microwave-assisted multicomponent polycondensation of amino acids. The compounds were evaluated for their antimicrobial activity against a panel of nine bacterial strains and five fungal strains. Compounds 26 and 27 were the most effective against Staphylococcus aureus ATCC 29213 reference strain with MIC values of 4 µg mL-1, and a methicillin-resistant Staphylococcus aureus (MRSA) isolate with MIC values of 8 µg mL-1. It was possible to infer that enantiomer (-)-26 was responsible for the antibacterial activity (MIC 4 µg mL-1) while (+)-26 had no activity. Furthermore, compound (-)-26 was able to impair S. aureus biofilm production and no significant cytotoxicity towards differentiated and non-differentiated SH-SY5Y cells was observed. Compounds 26, 28, and 29 showed a weak antifungal activity against Trichophyton rubrum clinical isolate with MIC 128 µg mL-1 and presented a synergistic effect with fluconazole.

Colangiopancreatografía retrógrada endoscópica en edad geriátrica / Endoscopic retrograde cholangiopancreatography in geriatric ages

En febrero de 2010 se introdujo la técnica colangiopancreatografía retrógrada endoscópica en la provincia de Matanzas, dando cobertura a los casos con esta indicación, y atendiendo a un importante grupo de pacientes en edad geriátrica. Por tal motivo, este trabajo describió los resultados después de cuatro años de trabajo con este proceder. Para ello se realizó un estudio descriptivo, transversal, en pacientes mayores de 65 años, quienes se realizaron el estudio en el servicio de Cirugía de Mínimo Acceso del Hospital Universitario Clínico Quirúrgico “Comandante Faustino Pérez Hernández”, en el período de febrero 2010 a febrero de 2015. La población de estudio quedó conformada por 147 pacientes, que cumplieron con los criterios de inclusión con 157 informes de colangiopancreatografía retrógrada endoscópica. Se analizaron variables demográficas, relacionadas con aspectos técnicos, y clínicas. Como resultados predominó el sexo femenino, con 51,7 %; el grupo etareo más frecuente fue de 65 a 69 años, con 64,63 %. La efectividad en la canulación fue del 97,28 %, realizándose precorte solo en 5,59 %; el diagnóstico más frecuente fue litiasis coledociana (38,16 %) y el 80,13 % requirió intervención terapéutica. Se presentaron complicaciones en 7,48 % y la más frecuente fue la colangitis aguda (3,40 %). La mortalidad fue de 1,36 %. La comparación con otros estudios realizados, permitió concluir que los resultados son similares a los realizados en poblaciones en general, independientemente de la edad, resultando ser un proceder seguro en edades geriátricas, siendo la colangiopancreatografía retrógrada endoscópica un arma con gran utilidad diagnóstica y terapéutica.

The cholangiopancratographic technique was introduced in February 2010 in the province of Matanzas, covering the cases with that indication, and attending to an important group of patients in geriatric age. The aim of the current work was describing the results of four years work with this procedure. For that, a cross-sectional, descriptive study was carried out in patients older than 65 years who were studied at the Minimal Access Surgery service of the University Clinical Surgical Hospital “Comandante Faustino Pérez Hernández”, in the period February 2010-February 2015. The studied population was formed by 147 patients, fulfilling the inclusion criteria, with 157 informs of endoscopic retrograde cholangiopancreatography. Clinical and demographic variables, related to technical aspects, were analyzed. The outcomes were the following: the female gender predominated, with 51.7 %; the most frequently found age group was the one grouping patients aged 65-69 years, with 64.63 %. The cannulation effectiveness was 97.28 %, making pre-cut only in 5.59 % of the cases; the most frequent diagnosis was choledochal lithiasis (38.16 %) and 80.13 % required therapeutic intervention. Complications were in 7.48 % of the cases, and the most frequent one was acute cholangitis (3.40 %). Mortality was 1.36 %. The comparison with other studies allowed concluding that the outcomes are similar to the ones obtained in studies carried out in general populations, independently of age, being a secure procedure in geriatric ages. The endoscopic retrograde cholangiopancreatography is a tool of great diagnostic and therapeutic usefulness.

Colangiopancreatografía retrógrada endoscópica en edad geriátrica / Endoscopic retrograde cholangiopancreatography in geriatric ages

En febrero de 2010 se introdujo la técnica colangiopancreatografía retrógrada endoscópica en la provincia de Matanzas, dando cobertura a los casos con esta indicación, y atendiendo a un importante grupo de pacientes en edad geriátrica. Por tal motivo, este trabajo describió los resultados después de cuatro años de trabajo con este proceder. Para ello se realizó un estudio descriptivo, transversal, en pacientes mayores de 65 años, quienes se realizaron el estudio en el servicio de Cirugía de Mínimo Acceso del Hospital Universitario Clínico Quirúrgico “Comandante Faustino Pérez Hernández”, en el período de febrero 2010 a febrero de 2015. La población de estudio quedó conformada por 147 pacientes, que cumplieron con los criterios de inclusión con 157 informes de colangiopancreatografía retrógrada endoscópica. Se analizaron variables demográficas, relacionadas con aspectos técnicos, y clínicas. Como resultados predominó el sexo femenino, con 51,7 por ciento; el grupo etareo más frecuente fue de 65 a 69 años, con 64,63 por ciento. La efectividad en la canulación fue del 97,28 por ciento, realizándose precorte solo en 5,59 por ciento; el diagnóstico más frecuente fue litiasis coledociana (38,16 por ciento) y el 80,13 por ciento requirió intervención terapéutica. Se presentaron complicaciones en 7,48 por ciento y la más frecuente fue la colangitis aguda (3,40 por ciento). La mortalidad fue de 1,36 por ciento. La comparación con otros estudios realizados, permitió concluir que los resultados son similares a los realizados en poblaciones en general, independientemente de la edad, resultando ser un proceder seguro en edades geriátricas, siendo la colangiopancreatografía retrógrada endoscópica un arma con gran utilidad diagnóstica y terapéutica(AU)

The cholangiopancratographic technique was introduced in February 2010 in the province of Matanzas, covering the cases with that indication, and attending to an important group of patients in geriatric age. The aim of the current work was describing the results of four years work with this procedure. For that, a cross-sectional, descriptive study was carried out in patients older than 65 years who were studied at the Minimal Access Surgery service of the University Clinical Surgical Hospital “Comandante Faustino Pérez Hernández”, in the period February 2010-February 2015. The studied population was formed by 147 patients, fulfilling the inclusion criteria, with 157 informs of endoscopic retrograde cholangiopancreatography. Clinical and demographic variables, related to technical aspects, were analyzed. The outcomes were the following: the female gender predominated, with 51,7 percent; the most frequently found age group was the one grouping patients aged 65-69 years, with 64,63 percent. The cannulation effectiveness was 97,28 percent, making pre-cut only in 5,59 percent of the cases; the most frequent diagnosis was choledochal lithiasis (38,16 percent) and 80,13 percent required therapeutic intervention. Complications were in 7,48 percent of the cases, and the most frequent one was acute cholangitis (3,40 percent). Mortality was 1,36 percent. The comparison with other studies allowed concluding that the outcomes are similar to the ones obtained in studies carried out in general populations, independently of age, being a secure procedure in geriatric ages. The endoscopic retrograde cholangiopancreatography is a tool of great diagnostic and therapeutic usefulness(AU)