Determination and comparison of the chemical composition of Calendula L. species growing in Continental Portugal

11th National Meeting of Organic Chemistry and 4th Meeting of Therapeutic Chemistry.The flora of Continental Portugal includes three species of Calendula L. (Calendula officinalis L., C. arvensis L., C. suffruticosa Vahl), one of which comprises three subspecies (C. suffruticosa subsp. algarbiensis (Boiss.) Nyman, C. suffruticosa subsp. lusitanica (Boiss.) Ohle and C. suffruticosa subsp. cinerea (Ohle) P.Silveira & A.C.Gonç.). C. officinalis is recognised for its medical properties and its chemical composition has been widely studied (Muley, B. P., et al. J. Pharm. Res. 2009, 8, 455; Safdar, W., et al. Int. J. Cell Mol. Biol. 2010, 1, 108). Nevertheless, little is known about the chemical composition of C. arvensis and even less regarding the different subspecies of C. suffruticosa. Therefore, the present study aims the elucidation of these plants’ chemical composition and to compare and identify differences and/or similarities among them. […].Instituto da Conservação da Natureza e das Florestas; Universidade de Aveiro; FCT / MEC (FCT UID / QUI / 00062/2013); UEM CESAM (UID / AMB / 50017); FEDER (Acordo de Parceria PT2020).info:eu-repo/semantics/publishedVersio

BASHY Dye Platform Enables the Fluorescence Bioimaging of Myelin Debris Phagocytosis by Microglia during Demyelination

Multiple sclerosis (MS) is a demyelinating disease of the central nervous system that is characterized by the presence of demyelinated regions with accumulated myelin lipid debris. Importantly, to allow effective remyelination, such debris must be cleared by microglia. Therefore, the study of microglial activity with sensitive tools is of great interest to better monitor the MS clinical course. Using a boronic acid-based (BASHY) fluorophore, specific for nonpolar lipid aggregates, we aimed to address BASHY’s ability to label nonpolar myelin debris and image myelin clearance in the context of demyelination. Demyelinated ex vivo organotypic cultures (OCSCs) and primary microglia cells were immunostained to evaluate BASHY’s co-localization with myelin debris and also to evaluate BASHY’s specificity for phagocytosing cells. Additionally, mice induced with experimental autoimmune encephalomyelitis (EAE) were injected with BASHY and posteriorly analyzed to evaluate BASHY+ microglia within demyelinated lesions. Indeed, in our in vitro and ex vivo studies, we showed a significant increase in BASHY labeling in demyelinated OCSCs, mostly co-localized with Iba1-expressing amoeboid/phagocytic microglia. Most importantly, BASHY’s presence was also found within demyelinated areas of EAE mice, essentially co-localizing with lesion-associated Iba1+ cells, evidencing BASHY’s potential for the in vivo bioimaging of myelin clearance and myelin-carrying microglia in regions of active demyelination.This work was supported by Grant for Multiple Sclerosis Innovation-Merck Serono to AF, Young Investigator’s Projects for Collaborative Cross-disciplinary Studies from iMed.ULisboa to A.F. and F.S.; PTDC/QUI-QOR/29967/2017 and LISBOA-01-0145-FEDER-029967 to P.G.; the Spanish Ministry of Science, Innovation, and Universities (CTQ2017-89832-P for U.P.); the PhD grant 2021.04911.BD from Fundação para a Ciência e Tecnologia, Portugal (FCT) to M.V.P.; and in part by UIDB/04138/2020 and UIDP/04138/2020-from FCT to iMed.ULisboa

Methodology for Abrasive Wear Evaluation in Elevator Stage Centrifugal Pump Impellers

The abrasion resistance of the impellers depends on the characteristics of the materials used in their manufacture. In this work, a methodology is proposed for the evaluation of the abrasive wear of the plates of the centrifugal pump impellers, used in the gross water infrastructure station (GWIS) of sedimentary rivers, due to the sediment load variation and the river fluviometric dimension. In order to determine the wear mode and the relationship of the material-specific wear coefficient (K), due to the sediment concentration, a rotating ball abrasometer test was performed on SAE 8620, gray cast iron (GCI), and nodular cast iron (NCI), used in the manufacture of impellers. As an abrasive suspension, the concentrations of 1, 2, 3, 5, and 10 g L−1 of sediment were used in distilled water. The wear volume as a function of the relative velocity of the mixture in relation to the impeller blades was estimated mathematically. The results show that: i) The abrasive capacity of the sediments in different concentrations; ii) SAE 8620 steel was more resistant to abrasive wear; and iii) the rotational control of the pump as a function of sediment concentration and river level showed the possibility of reducing wear by 30%

Ausência da perda de heterozigose do PRKAR1A em células capturadas por microdissecção a laser de tecido de nódulo pigmentoso adrenocortical de um paciente com complexo de Carney causado por uma nova mutação nonsense

OBJECTIVE: Primary pigmented nodular adrenocortical disease (PPNAD) is the main endocrine manifestation of Carney complex, a multiple neoplasia syndrome caused by PRKAR1A gene mutations. The presence of PRKAR1A loss of heterozygosity (LOH) in adrenocortical tumorigenesis remains controversial. The aim of the present study is to investigate the presence of PRKAR1A LOH in adrenocortical cells in a patient with Carney complex. METHODS: The LOH was investigated using a PRKAR1A informative intragenic marker by GeneScan software analysis in DNA obtained from laser-captured microdissected cells of several adrenal nodules. Patients: A young adult male patient with Carney complex and his family were studied. RESULTS: A novel heterozygous mutation (p. Y21X) was identified at PRKAR1A in blood DNA of the male proband and his relatives. No PRKAR1A LOH was evidenced in the laser-captured microdissected cells from PPNAD tissue by different methodologies. CONCLUSION: We identified a new PRKAR1A nonsense mutation and in addition we did not evidence PRKAR1A LOH in laser-captured nodules cells, suggesting that adrenocortical tumorigenesis in PPNAD may occurs apart from the second hit.OBJETIVO: A doença adrenocortical nodular pigmentosa primária (PPNAD) é uma das manifestações do complexo de Carney, uma neoplasia endócrina múltipla causada por mutações no PRKAR1A. A perda de heterozigose (LOH) do PRKAR1A na tumorigenese adrenal permanece controversa dada à possibilidade de contaminação com o tecido normal. Nosso objetivo foi investigar a presença de LOH no PRKAR1A a partir de células do nódulo adrenal de um paciente com complexo de Carney. MÉTODOS: A pesquisa da LOH do PRKAR1A foi realizada através do estudo de um marcador intragênico em DNA de células do nódulo adrenal microdissecadas a laser, evitando contaminação com o tecido normal. Pacientes: Um paciente com PPNAD e cinco familiares foram estudados. RESULTADOS: A nova mutação (p. Y21X) foi identificada no PRKAR1A sem evidência de LOH no tecido adrenal. CONCLUSÃO: Identificamos uma nova mutação no PRKAR1A e não evidenciamos LOH nas células dos nódulos adrenocorticais, sugerindo que a PPNAD possa ocorrer na ausência de um segundo evento molecular

Proof-of-Concept Study of Multifunctional Hybrid Nanoparticle System Combined with NIR Laser Irradiation for the Treatment of Melanoma

Supplementary Materials - available online at https://www.mdpi.com/article/10.3390/biom11040511/s1The global impact of cancer emphasizes the importance of developing innovative, effective and minimally invasive therapies. In the context of superficial cancers, the development of a multifunctional nanoparticle-based system and its in vitro and in vivo safety and efficacy characterization are, herein, proposed as a proof-of-concept. This multifunctional system consists of gold nanoparticles coated with hyaluronic and oleic acids, and functionalized with epidermal growth factor for greater specificity towards cutaneous melanoma cells. This nanoparticle system is activated by a near-infrared laser. The characterization of this nanoparticle system included several phases, with in vitro assays being firstly performed to assess the safety of gold nanoparticles without laser irradiation. Then, hairless immunocompromised mice were selected for a xenograft model upon inoculation of A375 human melanoma cells. Treatment with near-infrared laser irradiation for five minutes combined with in situ administration of the nanoparticles showed a tumor volume reduction of approximately 80% and, in some cases, led to the formation of several necrotic foci, observed histologically. No significant skin erythema at the irradiation zone was verified, nor other harmful effects on the excised organs. In conclusion, these assays suggest that this system is safe and shows promising results for the treatment of superficial melanoma.The authors would like to thank to Fundação para a Ciência e Tecnologia (FCT) for the essential financial support under the project’s references PTDC/BBB-BMC/0611/2012, UIDB/00645/2020, UIDB/04138/2020 and UIDP/04138/2020 as well as for the PhD fellowships SFRH/BD/148044/2019 and SFRH/BD/147306/2019.info:eu-repo/semantics/publishedVersio

Preclinical pharmacokinetics and biodistribution of anticancer dinuclear Palladium(II)-Spermine Complex (Pd2Spm) in mice

Palladium-based compounds are regarded as potential analogs to platinum anticancer drugs with improved properties. The present study assessed the pharmacokinetics and biodistribution of a dinuclear palladium(II)-spermine chelate (Pd2Spm), which has previously been shown to possess promising in vitro activity against several therapy-resistant cancers. Using inductively coupled plasma-mass spectrometry, the kinetic profiles of palladium/platinum in serum, serum ultrafiltrate and tissues (kidney, liver, brain, heart, lungs, ovaries, adipose tissue and mammary glands) were studied in healthy female Balb/c mice after a single intraperitoneal bolus injection of Pd2Spm (3 mg/kg bw) or cisplatin (3.5 mg/kg bw) between 0.5 and 48 h post-injection. Palladium in serum exhibited biphasic kinetics with a terminal half-life of 20.7 h, while the free palladium in serum ultrafiltrate showed a higher terminal half-life than platinum (35.5 versus 31.5 h). Palladium was distributed throughout most of the tissues except for the brain, with the highest values in the kidney, followed by the liver, lungs, ovaries, adipose tissue and mammary glands. The in vitro cellular accumulation was also evaluated in breast cancer cells, evidencing a passive diffusion as a mechanism of Pd2Spm’s cellular entry. This study reports, for the first time, the favorable pharmacokinetics and biodistribution of Pd2Spm, which may become a promising pharmacological agent for cancer treatmentinfo:eu-repo/semantics/publishedVersio