A comparison between pure active pharmaceutical ingredients and therapeutic deep eutectic solvents: solubility and permeability studies

THEDES, so called therapeutic deep eutectic solvents are here defined as a mixture of two components, which at a particular molar composition become liquid at room temperature and in which one of them is an active pharmaceutical ingredient (API). In this work, THEDES based on menthol complexed with three different APIs, ibuprofen (ibu), BA (BA) and phenylacetic acid (PA), were prepared. The interactions between the components that constitute the THEDES were studied by NMR, confirming that the eutectic system is formed by H-bonds between menthol and the API. The mobility of the THEDES components was studied by PFGSE NMR spectroscopy. It was determined that the self-diffusion of the species followed the same behavior as observed previously for ionic liquids, in which the components migrate via jumping between voids in the suprastructure created by punctual thermal fluctuations. The solubility and permeability of the systems in an isotonic solution was evaluated and a comparison with the pure APIs was established through diffusion and permeability studies carried out in a Franz cell. The solubility of the APIs when in the THEDES system can be improved up to 12 fold, namely for the system containing ibu. Furthermore, for this system the permeability was calculated to be 14 105 cm/s representing a 3 fold increase in comparison with the pure API. With the exception of the systems containing PA an increase in the solubility, coupled with an increase in permeability was observed. In this work, we hence demonstrate the efficiency of THEDES as a new formulation for the enhancement of the bioavailability of APIs by changing the physical state of the molecules from a solid dosage to a liquid system.he European Union’s Seventh Framework Programme (FP7/2007- 2013) under grant agreement nREGPOT-CT2012-316331- POLARIS and from Project ‘‘Novel smart and biomimetic materials for innovative regenerative medicine approaches (Ref.: RL1 - ABMR - NORTE-01-0124-FEDER-000016)” co-financed by North Portugal Regional Operational Programme (ON.2 – O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF). Funding was also provided by Fundação para a Ciência e a Tecnologia, through contracts LAQV-REQUIMTE: UID/QUI/50006/2013 and UCIBIOREQUIMTE: UID/Multi/04378/2013. This work was co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01- 0145-FEDER-007728).info:eu-repo/semantics/publishedVersio

A common yardstick to measure the effects of different extreme climatic events on soil arthropod community composition using time-series data

Extreme climatic events can have profound effects on ecosystems. Climate change is causing an increase in the frequency and intensity of extreme events, which raises the probability that natural ecosystems will be exposed to subsequent or simultaneous extreme events. Exposure to multiple extreme events may involve very different abiotic stressors, which makes it hard to compare their consequences for more than a single or a few species, limiting our knowledge of community level effects. Here, we propose a novel approach to assess how different abiotic stressors influenced soil arthropod communities in a coastal ecosystem, using a species abundance time series of more than 2 years. We first correlated shifts in community composition with the climatic conditions preceding the sampling dates. Temperature was found to be the most important factor influencing community composition in both locations. Based on local meteorological data, we then defined five types of extreme events which occurred during our monitoring period: heat, cold, drought, heavy precipitation, and high sea water levels. To compare the effect of different types of extreme climatic events on community composition, we calculated the multidimensional Hellinger distance between two subsequent sampling dates, which is a measure for the compositional dissimilarity between communities. Extreme events were expected to result in a larger change in community composition between sampling dates and thus a larger Hellinger distance. However, no significant difference in Hellinger distance was found for intervals with or without extreme events in three out of the four location-vegetation combinations. In the saline location with an open vegetation type we found that Hellinger's distance was reduced when extreme events had occurred, which is discussed in the light of the buffering potential of different vegetation types. Our study illustrates the novel use of an established method from the community ecological toolbox to facilitate direct comparison of different types of extreme climatic events on community composition. We highly encourage other ecologists with long-term monitoring datasets to perform similar analyses and test the general applicability of this method

Discards for Southern Hake Stock, a First Approach to Iberian Data

This document presents sampling levels obtained with Portuguese and Spanish Discard Sampling Programmes, both included in National Sampling Programmes, and some preliminary results for the Iberian Southern Hake discards. These programs consist on onboard-observer sampling schemes, with co-operative vessels, quasirandom selected, in the ICES Division VIIIc and IXa. The methodology used to estimate hake discards for Portugal and Spain since 2003 was based on the one proposed in the “Workshop on Discard Sampling Methodology and Raising Procedures” (PGCCDBS, Denmark, 2003). Spain presents results for the years 1994, 1997, 1999, 2000, 2003 and 2004. In the Portuguese Program, 2004 is the first year with an analysis of discard data. The number of trips sampled by the Spanish program for 2004 was 53, distributed by three trawl fleets- BACA trawl, Pair trawl and WHVO trawl. Portuguese onboard-observers sampled 40 trips distributed by Crustacean and Fish trawl fleets. The estimated percentages of discarded hake in relation to total catch were of 19,9 (C.V. = 22.6%) and 46,7 (C.V. = 72.1%) for the Spanish and Portuguese fleets respectively. The confidence interval for the Portuguese discard estimate is very large which denotes the very low precision of the obtained value. A co-operation in the analysis of this kind of data, between the two countries, could be the next step, since a standardisation of the methods would be of great importance for a realistic comparison of results obtained from both programs

A study protocol to evaluate the relationship between outdoor air pollution and pregnancy outcomes

<p>Abstract</p> <p>Background</p> <p>The present study protocol is designed to assess the relationship between outdoor air pollution and low birth weight and preterm births outcomes performing a semi-ecological analysis. Semi-ecological design studies are widely used to assess effects of air pollution in humans. In this type of analysis, health outcomes and covariates are measured in individuals and exposure assignments are usually based on air quality monitor stations. Therefore, estimating individual exposures are one of the major challenges when investigating these relationships with a semi-ecologic design.</p> <p>Methods/Design</p> <p>Semi-ecologic study consisting of a retrospective cohort study with ecologic assignment of exposure is applied. Health outcomes and covariates are collected at Primary Health Care Center. Data from pregnant registry, clinical record and specific questionnaire administered orally to the mothers of children born in period 2007-2010 in Portuguese Alentejo Litoral region, are collected by the research team. Outdoor air pollution data are collected with a lichen diversity biomonitoring program, and individual pregnancy exposures are assessed with spatial geostatistical simulation, which provides the basis for uncertainty analysis of individual exposures. Awareness of outdoor air pollution uncertainty will improve validity of individual exposures assignments for further statistical analysis with multivariate regression models.</p> <p>Discussion</p> <p>Exposure misclassification is an issue of concern in semi-ecological design. In this study, personal exposures are assigned to each pregnant using geocoded addresses data. A stochastic simulation method is applied to lichen diversity values index measured at biomonitoring survey locations, in order to assess spatial uncertainty of lichen diversity value index at each geocoded address. These methods assume a model for spatial autocorrelation of exposure and provide a distribution of exposures in each study location. We believe that variability of simulated exposure values at geocoded addresses will improve knowledge on variability of exposures, improving therefore validity of individual exposures to input in posterior statistical analysis.</p

C2-phytoceramide perturbs lipid rafts and cell integrity in Saccharomyces cerevisiae in a sterol-dependent manner

Specific ceramides are key regulators of cell fate, and extensive studies aimed to develop therapies based on ceramide-induced cell death. However, the mechanisms regulating ceramide cytotoxicity are not yet fully elucidated. Since ceramides also regulate growth and stress responses in yeast, we studied how different exogenous ceramides affect yeast cells. C2-phytoceramide, a soluble form of phytoceramides, the yeast counterparts of mammalian ceramides, greatly reduced clonogenic survival, particularly in the G2/M phase, but did not induce autophagy nor increase apoptotic markers. Rather, the loss of clonogenic survival was associated with PI positive staining, disorganization of lipid rafts and cell wall weakening. Sensitivity to C2-phytoceramide was exacerbated in mutants lacking Hog1p, the MAP kinase homolog of human p38 kinase. Decreasing sterol membrane content reduced sensitivity to C2-phytoceramide, suggesting sterols are the targets of this compound. This study identified a new function of C2-phytoceramide through disorganization of lipid rafts and induction of a necrotic cell death under hypo-osmotic conditions. Since lipid rafts are important in mammalian cell signaling and adhesion, our findings further support pursuing the exploitation of yeast to understand the basis of synthetic ceramides' cytotoxicity to provide novel strategies for therapeutic intervention in cancer and other diseases.This work was supported by Fundacao para a Ciencia e Tecnologia through projects PTDC/BIA-BCM/69448/2006 and PEst-C/BIA/UI4050/2011, and fellowships to A. P. (SFRH/BPD/65003) and F. A. (SFRH/BD/80934/2011), as well as by FEDER through POFC - COMPETE. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Validation of the questionnaire on beliefs about medication with type 2 diabetic patients

O presente trabalho teve como objectivo validar o Questionário Crenças sobre a Medicação, que avalia Crenças Gerais e Crenças Específicas, estudando suas propriedades psicométricas em uma amostra de 387 pacientes diabéticos tipo 2. O estudo de validade para as Crenças Gerais revelou uma solução de um factor, com um alfa de 0,76, e para as Crenças Específicas, dois factores – Necessidades e Preocupações –, com um alfa de 0,77 e 0,69 respectivamente. Quanto à validade de constructo, verificou-se uma relação entre as Crenças Gerais e a subescala Necessidades das Crenças Específicas com Adesão à Medicação, avaliada pela Escala de Avaliação de Aderência Médica. O instrumento apresenta boas qualidades psicométricas para ser utilizado em pacientes diabéticos tipo 2.The present paper focused on the validation of the Questionnaire on Beliefs about Medication, which assesses both General Beliefs and Specific Beliefs. The psychometric properties of the instrument were analyzed on a sample of 387 type 2 diabetic patients. The validity study for General Beliefs found a unifactorial solution, with an alpha of .76, and for Specific Beliefs, a two-factor solution – Necessities and Concern –, with an alpha of .77 and .69, respectively. In terms of construct validity, a relationship between General Beliefs, subscale Necessities from Specific Beliefs, and adherence to medication, as evaluated by Medical Adherence Rating Scale, was found. The instrument presents good psychometric qualities to be used in type 2 diabetic patients.Fundação para a Ciência e Tecnologia (FCT

Photobiomodulation reduces the cytokine storm syndrome associated with Covid-19 in the zebrafish model

Although the exact mechanism of the pathogenesis of COVID-19 is not fully understood, oxidative stress and the release of pro-inflammatory cytokines have been highlighted as playing a vital role in the pathogenesis of the disease. In this sense, alternative treatments are needed to reduce the inflammation caused by COVID-19. Therefore, this study aimed to investigate the potential effect of red PBM as an attractive therapy to downregulate the cytokine storm caused by COVID-19 from a zebrafish model. RT-PCR analyses and protein-protein interaction prediction among SARS-CoV-2 and Danio rerio proteins showed that rSpike was responsible for generating systemic inflammatory processes with significantly increased pro-inflammatory (il1b, il6, tnfa, and nfkbiab), oxidative stress (romo1) and energy metabolism (slc2a1a, coa1) mRNA markers, with a pattern like those observed in COVID-19 cases in humans. On the other hand, PBM treatment decreased the mRNA levels of these pro-inflammatory and oxidative stress markers compared with rSpike in various tissues, promoting an anti-inflammatory response. Conversely, PBM promotes cellular and tissue repair of injured tissues and significantly increases the survival rate of rSpike-inoculated individuals. Additionally, metabolomics analysis showed that the most impacted metabolic pathways between PBM and the rSpike-treated groups were related to steroid metabolism, immune system, and lipids metabolism. Together, our findings suggest that the inflammatory process is an incisive feature of COVID-19, and red PBM can be used as a novel therapeutic agent for COVID-19 by regulating the inflammatory response. Nevertheless, the need for more clinical trials remains, and there is a significant gap to overcome before clinical trials.publishedVersio

Evaluation of an Antimicrobial L-Amino Acid Oxidase and Peptide Derivatives from Bothropoides mattogrosensis Pitviper Venom

Healthcare-associated infections (HAIs) are causes of mortality and morbidity worldwide. The prevalence of bacterial resistance to common antibiotics has increased in recent years, highlighting the need to develop novel alternatives for controlling these pathogens. Pitviper venoms are composed of a multifaceted mixture of peptides, proteins and inorganic components. L-amino oxidase (LAO) is a multifunctional enzyme that is able to develop different activities including antibacterial activity. In this study a novel LAO from Bothrops mattogrosensis (BmLAO) was isolated and biochemically characterized. Partial enzyme sequence showed full identity to Bothrops pauloensis LAO. Moreover, LAO here isolated showed remarkable antibacterial activity against Gram-positive and -negative bacteria, clearly suggesting a secondary protective function. Otherwise, no cytotoxic activities against macrophages and erythrocytes were observed. Finally, some LAO fragments (BmLAO-f1, BmLAO-f2 and BmLAO-f3) were synthesized and further evaluated, also showing enhanced antimicrobial activity. Peptide fragments, which are the key residues involved in antimicrobial activity, were also structurally studied by using theoretical models. The fragments reported here may be promising candidates in the rational design of new antibiotics that could be used to control resistant microorganisms

Human Apolipoprotein A-I-Derived Amyloid: Its Association with Atherosclerosis

Amyloidoses constitute a group of diseases in which soluble proteins aggregate and deposit extracellularly in tissues. Nonhereditary apolipoprotein A-I (apoA-I) amyloid is characterized by deposits of nonvariant protein in atherosclerotic arteries. Despite being common, little is known about the pathogenesis and significance of apoA-I deposition. In this work we investigated by fluorescence and biochemical approaches the impact of a cellular microenvironment associated with chronic inflammation on the folding and pro-amyloidogenic processing of apoA-I. Results showed that mildly acidic pH promotes misfolding, aggregation, and increased binding of apoA-I to extracellular matrix elements, thus favoring protein deposition as amyloid like-complexes. In addition, activated neutrophils and oxidative/proteolytic cleavage of the protein give rise to pro amyloidogenic products. We conclude that, even though apoA-I is not inherently amyloidogenic, it may produce non hereditary amyloidosis as a consequence of the pro-inflammatory microenvironment associated to atherogenesis