Screening of Potential Stress Biomarkers in Sweat Associated with Sports Training

The authors acknowledge the financial support of Fundacao para a Ciencia e Tecnologia (FCT/MCTES), grant PTDC/SAU-SOC/28390/2017. Also, this work was supported by the Associate Laboratory for Green Chemistry - LAQV which is financed by national funds from FCT/MCTES (UIDB/50006/2020).Background: Intense and continuous physical training in sports is related with psychological and physiological stress, affecting the health and well-being of athletes. The development of non-invasive sampling methodologies is essential to consider sweat as a potential biological fluid for stress biomarker assessment. In the current work, the identification in sweat samples of potential molecules that may be used as stress biomarkers was pursued. Methods: A sweat pool sample from football players after a 90-min intense training game was studied. Results: An analysis method using liquid chromatography with detection by tandem mass spectrometry (LC-MSMS) to attain a screening profile of sweat composition is presented. The major focus was on neurotransmitters (e.g. monoamines and metabolites) and other biological molecules related with physical training, such as precursors of biogenic amines (phenylaniline, tyrosine, etc.). Conclusions: This study allowed the identification of small biomolecules, neurotransmitters and other related molecules in sweat that are potentially associated with stress conditions. The developed methodology intends to contribute to the assessment and study of physical and psychological stress biomarkers related with intense sports using non-invasive methods.publishersversionpublishe

A Novel Approach for Bisphosphonates: Ionic Liquids and Organic Salts from Zoledronic Acid

Novel ionic liquids and organic salts based on mono- or dianionic zoledronate and protonated superbases, choline and n-alkylmethylimidazolium cations, were prepared and characterized by spectroscopic and thermal analyses. Most of the prepared salts display amorphous structures and very high solubility in water and saline solutions, especially the dianionic salts. Among the zoledronate-based ionic compounds, those containing choline [Ch] and methoxyethylmethylimidazolium [C3 OMIM] cations appear to have significant cytotoxicity against human osteosarcoma cells (MG63) and low toxicity toward healthy skin fibroblast cells. Because osteosarcoma is a bone pathology characterized by an increase in bone turnover rate, the results presented herein may be a promising starting point for the development of new ionic pharmaceutical drugs against osteosarcoma.info:eu-repo/semantics/publishedVersio

Mito-nuclear sequencing is paramount to correctly identify sympatric hybridizing fishes

Background. Hybridization may drive speciation and erode species, especially when intrageneric sympatric species are involved. Five sympatric Luciobarbus species—Luciobarbus sclateri (Günther, 1868), Luciobarbus comizo (Steindachner, 1864), Luciobarbus microcephalus (Almaça, 1967), Luciobarbus guiraonis (Steindachner, 1866), and Luciobarbus steindachneri (Almaça, 1967)—are commonly identified in field surveys by diagnostic morphological characters. Assuming that i) in loco identification is subjective and observer-dependent, ii) there is previous evidence of interspecific hybridization, and iii) the technical reports usually do not include molecular analyses, our main goal was to assess the concordance between in loco species identification based on phenotypic characters with identifications based on morphometric indices, mtDNA only, and a combination of mito-nuclear markers. Materials and methods. Specimens of Luciobarbus from six Guadiana River sub-basins were collected and sequenced for the cytochrome b and beta-actin genes. For comparative purposes, samples of Luciobarbus from other 12 river basins were also used. Four levels of taxonomical identification were conducted based on: identification made in the field (in loco identification), cytb gene only, beta-actin gene only, and mito-nuclear combined genomes. Results. Results showed that interspecific hybridization seems to be high (around 41%) and likely favoured by non-random mating and the loss of fluvial connectivity. About 34% of the hybrids showed mito-nuclear discordance. Misidentifications were frequent when only phenotypic characters are considered, and the use of a single mitochondrial gene is not sufficient: the use of two mito-nuclear markers showed that around 82% of the in loco identifications based on the phenotype were not correct. Conclusion. Incorrect species assignment likely generated biased results in previous studies on the biology and ecology of Guadiana barbels and in the assignment of conservation status and, consequently, on the establishment of conservation management measuresinfo:eu-repo/semantics/publishedVersio

Migração celular distinta induzida por Leishmania infantum chagasi e saliva de Lutzomyia longipalpis em um modelo de pool hemorrágico

Recruitment of a specific cell population after Leishmania infection can influence the outcome of the disease. Cellular migration in response to Leishmania or vector saliva has been reported in air pouch model, however, cellular migration induced by Leishmania associated with host's blood and vector saliva in this model has not been described. Herein we investigated cellular migration into air pouch of hamster after stimulation with combination of L. chagasi and host's blood and Lutzomyia longipalpis saliva. Migration induced by saliva was 3-fold more than those induced by L. chagasi alone. Additionally, L. chagasi associated with blood and saliva induced significantly even more leukocytes into air pouch than Leishmania alone. L. chagasi recruited a diverse cell population; however, most of these cells seem to have not migrated to the inflammatory exudate, remaining in the pouch lining tissue. These results indicate that L. chagasi can reduce leukocyte accumulation to the initial site of infection, and when associated with vector saliva in the presence of blood components, increase the influx of more neutrophils than macrophages, suggesting that the parasite has developed a strategy to minimize the initial inflammatory response, allowing an unlimited progression within the host. This work reinforces the importance of studies on the salivary components of sand fly vectors of leishmaniasis in the transmission process and the establishment of the infection.O recrutamento de uma população de células específicas após infecção por Leishmania pode influenciar o resultado da doença. A migração celular em resposta a Leishmania ou saliva do vetor tem sido reportada utilizando o modelo da bolsa de ar subcutânea, entretanto, a migração celular induzida por Leishmania associada com o sangue do hospedeiro e saliva do vetor neste modelo ainda não foi descrita. Neste trabalho foi investigada a migração celular no modelo da bolsa de ar subcutânea em hamster após a estimulação com a combinação de L. chagasi, sangue do hospedeiro e saliva de Lutzomyia longipalpis. A migração induzida por saliva foi três vezes maior do que a induzida por L. chagasi sozinha. Adicionalmente, L. chagasi associada com sangue e saliva induziu significativamente ainda mais leucócitos no exsudato inflamatório do que o estímulo com Leishmania sozinha. L. chagasi recrutou uma população de células distintas, no entanto, a maioria dessas células parece não ter migrado para o exsudato inflamatório, permanecendo no tecido da bolsa de ar. Estes resultados indicam que L. chagasi pode reduzir o acúmulo de leucócitos para o local inicial da infecção e que quando associada à saliva do vetor e na presença de componentes do sangue aumenta o influxo de mais neutrófilos do que macrófagos, sugerindo que o parasito desenvolveu uma estratégia para minimizar a resposta inflamatória inicial, permitindo uma progressão ilimitada dentro do hospedeiro. Este trabalho reforça a importância de mais estudos sobre os componentes da saliva dos vetores das leishmanioses no processo de transmissão e no estabelecimento da infecção

Synthesis and antibacterial activity of ionic liquids and organic salts based on penicillin g and amoxicillin hydrolysate derivatives against resistant bacteria

Fundacao para a Ciencia e Tecnologia through projects (PTDC/QUI-QOR/32406/2017, PEst-C/LA0006/2013) and PTDC/BTM-SAL/29786/2017, and one contract under Investigador FCT (L.C.B.). ZP thanks to Fundacao para a Ciencia e a Tecnologia, MCTES, for the Norma transitoria DL 57/2016 Program Contract. This work was also supported by the Associate Laboratory for Green Chemistry, which is financed by national funds from FCT/MEC, UIDB/50006/2020 and co‐financed by the ERDF under the PT2020 Partnership Agreement (POCI‐01‐ 0145‐FEDER‐007265). The authors also thank Solchemar company for support.The preparation and characterization of ionic liquids and organic salts (OSILs) that contain anionic penicillin G [secoPen] and amoxicillin [seco-Amx] hydrolysate derivatives and their in vitro antibacterial activity against sensitive and resistant Escherichia coli and Staphylococcus aureus strains is reported. Eleven hydrolyzed β-lactam-OSILs were obtained after precipitation in moderate-to-high yields via the neutralization of the basic ammonia buffer of antibiotics with different cation hydroxide salts. The obtained minimum inhibitory concentration (MIC) data of the prepared compounds showed a relative decrease of the inhibitory concentrations (RDIC) in the order of 100 in the case of [C2OHMIM][seco-Pen] against sensitive S. aureus ATCC25923 and, most strikingly, higher than 1000 with [C16Pyr][seco-Amx] against methicillin-resistant Staphylococcus aureus (MRSA) ATCC 43300. These outstanding in vitro results showcase that a straightforward transformation of standard antibiotics into hydrolyzed organic salts can dramatically change the pharmaceutical activity of a drug, including giving rise to potent formulations of antibiotics against deadly bacteria strains.publishersversionpublishe

Synthesis and biological evaluation of Amphotericin B formulations based on organic salts and ionic liquids against Leishmania infantum

Nowadays, organic salts and ionic liquids (OSILs) containing active pharmaceutical ingredients (APIs) are being explored as drug delivery systems in modern therapies (OSILs-API). In that sense, this work is focused on the development of novel OSILs-API based on amphotericin B through an innovative procedure and the evaluation of the respective biological activity against Leishmania infantum. Several ammonium, methylimidazolium, pyridinium and phosphonium organic cations combined with amphotericin B as anion were synthesized in moderate to high yields and high purities by the water-reduced buffer neutralization method. All prepared compounds were characterized to confirm the desired chemical structure and the specific optical rotation ([α]D25) was also determined. The biological assays performed on L. infantum promastigotes showed increased activity against this parasitic disease when compared with the starting chloride forms and amphotericin B alone, highlighting [P6,6,6,14][AmB] as the most promising formulation. Possible synergism in the antiprotozoal activity was also evaluated for [P6,6,6,14][AmB], since it was proven to be the compound with the highest toxicity. This work reported a simple synthetic method, which can be applied to prepare other organic salts based on molecules containing fragile chemical groups, demonstrating the potential of these OSILs-AmB as possible agents against leishmaniasis.info:eu-repo/semantics/publishedVersio

Anti-DLL4 VNAR targeted nanoparticles for targeting of both tumour and tumour associated vasculature

Acknowledgements The authors acknowledge the Engineering and Physical Sciences Research Council (EPSRC) (S3802ASA) and the generous support of the Martin Family Foundation for funding the Ph.D. studentships of P. S. and A. L., respectively. This work was also partially funded through a US-Ireland R&D Partnership grant awarded by HSCNI (STL/5010/14), Medical Research Council UK (MC_PC_15013), and the Biotechnology and Biological Sciences Research Council (BBSRC) (BB/R009112/1).Peer reviewedPublisher PD

Bioactivity of Ionic Liquids Based on Valproate in SH-SY5Y Human Neuroblastoma Cell Line

This work received financial support from the PT national funds (FCT-MCTES, Fundação para a Ciência e Tecnologia and Ministério da Ciência, Tecnologia e Ensino Superior) through the project UIDB/50006/2020 | UIDP/50006/2020. A.F.M.S. and Ž.P. also acknowledge FCT-MCTES for the PhD grant (SFRH/BD/132551/2017) and Norma Transitória DL 57/2016 program contract, respectivelyThe search for alternative and effective therapies to fight cancer is one of the main goals of the pharmaceutical industry. Recently, ionic liquids (ILs) have emerged as potential therapeutic agents with antitumor properties. The goal of this study was to synthesize and evaluate the bioactivity of different ILs coupled with the active pharmaceutical ingredient (API) valproate (VPA) as an antitumor agent. The toxicity of the prepared ionic liquids was evaluated by the MTT cell metabolic assay in human neuroblastoma SH-SY5Y and human primary Gingival Fibroblast (GF) cell lines, in which they showed inhibitory effects during the study period. In addition, low cytotoxicity against GF cell lines was observed, suggesting that these compounds are not toxic to human cell lines. [C2OHDMiM][VPA] demonstrated an outstanding antitumor activity against SH-SY5Y and lower activity against the non-neoplastic GF line. The herein assessed compounds played an important role in the modulation of the signaling pathways involved in the cellular behavior. This work also highlights the potential of these ILs-API as possible antitumor agentspublishersversionpublishe

Experimental Tests and Preliminary Life Cycle Assessment

Funding Information: Thanks are also due to FCT for the project PTDC/BII-BIO/30884/2017. A.F.P. and M.M.M. thank FCT for funding through the Individual Call to Scientific Employment Stimulus (ref. 2020.01614.CEECIND/CP1596/CT0007 and CEEC-IND/02702/2017, respectively). O.D. is thankful for the Ph.D. grant supported by FCT (2021.05322.BD). Funding Information: This work was funded by Portuguese funds through Fundação para a Ciência e a Tecnologia (FCT/MCTES) in the framework of the projects UIDB/50006/2020, UIDP/50006/2020, and EXPL/BII-BIO/0436/2021 and also by BIP Proof 2022/2023 through UI-Transfer, a cofunding project by European Union through COMPETE 2020-Portugal 2020, CIQUP, Centro de Investigação em Química da Universidade do Porto (UIDB/00081/2020), and IMS, Institute of Molecular Sciences (LA/P/0056/2020). Publisher Copyright: © 2023 The Authors. Published by American Chemical Society.The control of the oxidative stability of biodiesel and blends of biodiesel with diesel is one of the major concerns of the biofuel industry. The oxidative degradation of biodiesel can be accelerated by several factors, and this is most critical in the so-called second generation biodiesel, which is produced from low-cost raw materials with lower environmental impacts. The addition of antioxidants is imperative to ensure the oxidative stability of biodiesel, and these are considered products of high commercial value. The antioxidants currently available on the market are from synthetic origin, so the existence/availability of alternative antioxidants of natural origin (less dependent on fossil sources) at a competitive price presents itself as a strong business opportunity. This work describes and characterizes a sustainable alternative to synthetic antioxidants used in the biodiesel market developed from extracts of vineyard pruning waste (VPW), which are naturally rich in phenolic compounds with antioxidant properties. A hydrothermal extraction process was applied as a more efficient and sustainable technology than the conventional one with the potential of the extracts as antioxidant additives in biodiesel evaluated in Rancitech equipment. The VPW extract showed comparable antioxidant activity as the commercial antioxidant butylated hydroxytoluene (BHT) typically used in biodiesel. The stability of the biodiesel is dependent from the amount of the extract added. Further, for the first time, the assessment of the environmental impacts of using natural extracts to control the oxidative stability of biodiesel in the production process is also discussed as a key factor of the process environmental sustainability.publishersversionpublishe

Profile of opioid prescriptions for intubated and mechanically ventilated neonates

Objective: to identify factors that affect the decision of prescribing opioids for intubated and ventilated neonates Method: retrospective study of intubated and ventilated newborn infants for periods longer than one hour, admitted to the NICU from January 1995 to June 1997. During this period, 203 patients fulfilled inclusion criteria and data of 176 charts were reviewed. Charts were analyzed regarding demographic data, characteristics of analgesia and respiratory support, invasive procedures performed and clinical entities diagnosed during the period of mechanical ventilation. Discriminative analysis was used to understand factors that lead to opioid use by some of these patients. Results: Ninety-seven neonates received at least one dose of opioids during the period of mechanical ventilation. None of these patients was evaluated with pain scales, and in 63% of them we could not retrieve any reason for opioid prescription in their charts. Discriminative analysis showed that the main differences between groups were birthweight, gestational age, oxygenation index at intubation, and number of arterial sticks during the first 72 hours of mechanical ventilation. The most mature and heaviest neonates with a more severe respiratory insufficiency received opioid analgesia during ventilation. Conclusion: the decision to use opioids in intubated and ventilated neonates was based on the infants' aspect and respiratory status. It did not consider the pain these patients might be suffering and it was not based on the evaluation of pain.Objetivo: identificar fatores que levaram os médicos a prescreverem opióides a recém-nascidos em ventilação mecânica. Método: estudo retrospectivo de pacientes em ventilação mecânica por cânula traqueal, por mais de 1 hora, internados em UTI neonatal entre janeiro de 1995 a junho de 1997. Nesse período, 203 recém-nascidos preencheram o critério de inclusão, recuperando-se 176 prontuários. Os prontuários foram analisados quanto a dados demográficos, características da analgesia e do suporte ventilatório, procedimentos invasivos realizados e entidades mórbidas diagnosticadas durante o período de ventilação. Para entender os fatores que determinaram o uso da analgesia em parte dessa população, utilizou-se a análise discriminante. Resultados: Nos 97 pacientes que receberam > 1 dose de opióides durante a ventilação, a analgesia foi iniciada, em média, até 24 horas após o início da ventilação. As escalas de avaliação da dor não foram usadas em nenhum paciente e, em 63%, não havia relato do motivo para a analgesia. A análise discriminante mostrou que as variáveis que diferenciaram os grupos submetidos ou não à analgesia foram: peso ao nascer, idade gestacional, índice de oxigenação e número de punções arteriais. Os neonatos com maior chance de receberem alguma dose de opióide durante a ventilação foram os de peso mais elevado, idade gestacional mais avançada, índice de oxigenação mais acentuado no início da ventilação e maior necessidade de gasometrias, ou seja, os bebês mais maduros e com doença respiratória mais grave. Conclusão: os médicos não levam em conta a dor propriamente dita e nem a avaliam para decidirem pela necessidade de analgesia no neonato em ventilaçãoUniversidade Federal de São Paulo (UNIFESP) - Escola Paulista de Medicina (UNIFESP-EPM)UNIFESP-EPM Disciplina de Pediatria NeonatalUNIFESP-EPM Disciplina de BioestatísticaUNIFESP, - Escola Paulista de Medicina (UNIFESP-EPM)UNIFESP, EPM Disciplina de Pediatria NeonatalUNIFESP, EPM Disciplina de BioestatísticaSciEL