Identification of Marker Compounds and In Vitro Toxicity Evaluation of Two Portuguese Asphodelus Leaf Extracts

This article belongs to the Special Issue Discovery of Bioactive Ingredients from Natural Products III.The leaves of Asphodelus bento-rainhae subsp. bento-rainhae, an endemic Portuguese species, and Asphodelus macrocarpus subsp. macrocarpus have been used as food, and traditionally as medicine, for treating ulcers, urinary tract, and inflammatory disorders. The present study aims to establish the phytochemical profile of the main secondary metabolites, together with the antimicrobial, antioxidant and toxicity assessments of both Asphodelus leaf 70% ethanol extracts. Phytochemical screenings were conducted by the TLC and LC-UV/DAD-ESI/MS chromatographic technique, and quantification of the leading chemical classes was performed by spectrophotometric methods. Liquid-liquid partitions of crude extracts were obtained using ethyl ether, ethyl acetate, and water. For in vitro evaluations of antimicrobial activity, the broth microdilution method, and for the antioxidant activity, the FRAP and DPPH methods were used. Genotoxicity and cytotoxicity were assessed by Ames and MTT tests, respectively. Twelve known compounds including neochlorogenic acid, chlorogenic acid, caffeic acid, isoorientin, p-coumaric acid, isovitexin, ferulic acid, luteolin, aloe-emodin, diosmetin, chrysophanol, and β-sitosterol were identified as the main marker compounds, and terpenoids and condensed tannins were found to be the major class of secondary metabolites of both medicinal plants. The ethyl ether fractions demonstrated the highest antibacterial activity against all the Gram-positive microorganisms, (MIC value of 62 to 1000 µg/mL), with aloe-emodin as one of the main marker compounds highly active against Staphylococcus epidermidis (MIC value of 0.8 to 1.6 µg/mL). Ethyl acetate fractions exhibited the highest antioxidant activity (IC50 of 800 to 1200 µg/mL, respectively). No cytotoxicity (up to 1000 µg/mL) or genotoxicity/mutagenicity (up to 5 mg/plate, with/without metabolic activation) were detected. The obtained results contribute to the knowledge of the value and safety of the studied species as herbal medicines.This research was funded by the Foundation for Science and Technology (FCT, Portugal) through national funds FCT/MCTES to iMed.ULisboa (UIDP/04138/2020, UIDB/04138/2020) and MEtRICs (UIDP/04077/2020, UIDB/04077/2020) research projects, as well as doctoral scholarship (SFRH/BD/125310/2016) granted to the first author.info:eu-repo/semantics/publishedVersio

Structure of Hierridin C, Synthesis of Hierridins B and C, and Evidence for Prevalent Alkylresorcinol Biosynthesis in Picocyanobacteria

Small, single-celled planktonic cyanobacteria are ubiquitous in the world's oceans yet tend not to be perceived as secondary metabolite-rich organisms. Here we report the isolation and structure elucidation of hierridin C, a minor metabolite obtained from the cultured picocyanobacterium Cyanobium sp. LEGE 06113. We describe a simple, straightforward synthetic route to the scarcely produced hierridins that relies on a key regioselective halogenation step. In addition, we show that these compounds originate from a type III PKS pathway and that similar biosynthetic gene clusters are found in a variety of bacterial genomes, most notably those of the globally distributed picocyanobacteria genera Prochlorococcus, Cyanobium and Synechococcus.info:eu-repo/semantics/publishedVersio

Water and sodium intake habits and status of ultra-endurance runners during a multi-stage ultra-marathon conducted in a hot ambient environment: an observational field based study

<p>Abstract</p> <p>Background</p> <p>Anecdotal evidence suggests ultra-runners may not be consuming sufficient water through foods and fluids to maintenance euhydration, and present sub-optimal sodium intakes, throughout multi-stage ultra-marathon (MSUM) competitions in the heat. Subsequently, the aims were primarily to assess water and sodium intake habits of recreational ultra-runners during a five stage 225 km semi self-sufficient MSUM conducted in a hot ambient environment (T_max range: 32°C to 40°C); simultaneously to monitor serum sodium concentration, and hydration status using multiple hydration assessment techniques.</p> <p>Methods</p> <p>Total daily, pre-stage, during running, and post-stage water and sodium ingestion of ultra-endurance runners (UER, <it>n</it> = 74) and control (CON, <it>n</it> = 12) through foods and fluids were recorded on Stages 1 to 4 by trained dietetic researchers using dietary recall interview technique, and analysed through dietary analysis software. Body mass (BM), hydration status, and serum sodium concentration were determined pre- and post-Stages 1 to 5.</p> <p>Results</p> <p>Water (overall mean (SD): total daily 7.7 (1.5) L/day, during running 732 (183) ml/h) and sodium (total daily 3.9 (1.3) g/day, during running 270 (151) mg/L) ingestion did not differ between stages in UER (<it>p</it> < 0.001 <it>vs</it>. CON). Exercise-induced BM loss was 2.4 (1.2)% (<it>p</it> < 0.001). Pre- to post-stage BM gains were observed in 26% of UER along competition. Pre- and post-stage plasma osmolality remained within normal clinical reference range (280 to 303 mOsmol/kg) in the majority of UER (<it>p</it> > 0.05 <it>vs</it>. CON pre-stage). Asymptomatic hyponatraemia (<135 mmol/L) was evident pre- and post-stage in <it>n</it> = 8 UER, corresponding to 42% of sampled participants. Pre- and post-stage urine colour, urine osmolality and urine/plasma osmolality ratio increased (<it>p</it> < 0.001) as competition progressed in UER, with no change in CON. Plasma volume and extra-cellular water increased (<it>p</it> < 0.001) 22.8% and 9.2%, respectively, from pre-Stage 1 to 5 in UER, with no change in CON.</p> <p>Conclusion</p> <p>Water intake habits of ultra-runners during MSUM conducted in hot ambient conditions appear to be sufficient to maintain baseline euhydration levels. However, fluid over-consumption behaviours were evident along competition, irrespective of running speed and gender. Normonatraemia was observed in the majority of ultra-runners throughout MSUM, despite sodium ingestion under benchmark recommendations.</p

Early mobilisation in critically ill COVID-19 patients: a subanalysis of the ESICM-initiated UNITE-COVID observational study

Background Early mobilisation (EM) is an intervention that may improve the outcome of critically ill patients. There is limited data on EM in COVID-19 patients and its use during the first pandemic wave. Methods This is a pre-planned subanalysis of the ESICM UNITE-COVID, an international multicenter observational study involving critically ill COVID-19 patients in the ICU between February 15th and May 15th, 2020. We analysed variables associated with the initiation of EM (within 72 h of ICU admission) and explored the impact of EM on mortality, ICU and hospital length of stay, as well as discharge location. Statistical analyses were done using (generalised) linear mixed-effect models and ANOVAs. Results Mobilisation data from 4190 patients from 280 ICUs in 45 countries were analysed. 1114 (26.6%) of these patients received mobilisation within 72 h after ICU admission; 3076 (73.4%) did not. In our analysis of factors associated with EM, mechanical ventilation at admission (OR 0.29; 95% CI 0.25, 0.35; p = 0.001), higher age (OR 0.99; 95% CI 0.98, 1.00; p ≤ 0.001), pre-existing asthma (OR 0.84; 95% CI 0.73, 0.98; p = 0.028), and pre-existing kidney disease (OR 0.84; 95% CI 0.71, 0.99; p = 0.036) were negatively associated with the initiation of EM. EM was associated with a higher chance of being discharged home (OR 1.31; 95% CI 1.08, 1.58; p = 0.007) but was not associated with length of stay in ICU (adj. difference 0.91 days; 95% CI − 0.47, 1.37, p = 0.34) and hospital (adj. difference 1.4 days; 95% CI − 0.62, 2.35, p = 0.24) or mortality (OR 0.88; 95% CI 0.7, 1.09, p = 0.24) when adjusted for covariates. Conclusions Our findings demonstrate that a quarter of COVID-19 patients received EM. There was no association found between EM in COVID-19 patients' ICU and hospital length of stay or mortality. However, EM in COVID-19 patients was associated with increased odds of being discharged home rather than to a care facility. Trial registration ClinicalTrials.gov: NCT04836065 (retrospectively registered April 8th 2021)

Caracterização da atividade anti-malárica de derivados da quinazolina

A malária continua a ser uma das doenças parasitárias mais relevantes no mundo – segundo a OMS, houve 216 milhões de casos clínicos em 2016, sendo a doença parasitária que mais mortes provoca mundialmente. É causada por cinco espécies distintas do género Plasmodium que é transmitido ao ser humano pela picada do mosquito fêmea Anopheles spp. A resistência aos fármacos anti-maláricos em uso clínico bem como a inexistência de uma vacina eficaz constituem o principal obstáculo no controlo da doença. A rápida disseminação da resistência torna crucial a pesquisa e síntese de novos compostos com ação anti-malárica, capazes de atuar nos vários estadios de desenvolvimento do parasita. Este estudo tem como objetivo avaliar a atividade anti-malárica in vitro de P. falciparum de 18 novos compostos da subclasse de alcaloides da quinazolinona, calculando-se o valor de IC50. Os compostos foram sintetizados no Laboratório de Química Orgânica e Farmacêutica do Departamento de Ciências Químicas, da Faculdade de Farmácia da Universidade do Porto pelo grupo da Professora Doutora Maria Emília Sousa. Para os 3 compostos com melhor atividade biológica SL4C, SL5C e SL20C, foi avaliada a citotoxicidade em células de mamífero V79 através do ensaio MTT e calculado o Índice de Seletividade (IS), a hemotoxicidade (atividade hemolítica) e a capacidade de inibição da polimerização da hemozoína (β-hematina) in vitro. Os compostos alcaloides da quinazolinona SL4C, SL5C e SL20C demonstraram eficácia na inibição do crescimento de P. falciparum in vitro com IC50 ≤ 0,2 μM. Estes compostos não apresentaram atividade hemolítica in vitro. Não foi detetada a inibição da polimerização da hemozoína (β-hematina) in vitro dos compostos nas doses testadas. Os compostos não revelaram ser citotóxicos in vitro em células de mamífero não tumorais V79 (DL50 ≤ 14 μM), com um valor de IS ≥ 19. As quinazolinonas SL4C, SL5C e SL20C possuem caraterísticas com potencial para o desenvolvimento como novos fármacos anti-maláricos.Malaria continues to be one of the most important parasitic diseases in the world - according to the WHO, there were 216 million clinical cases in 2016, being the parasitic disease that causes more deaths worldwide. It is caused by five distinct species of the genus Plasmodium that is transmitted to humans by the bite of the female mosquito Anopheles spp. Resistance to antimalarial drugs in clinical use as well as the lack of an effective vaccine constitute the main obstacle in controlling the disease. The rapid spread of resistance makes it crucial to research and synthesize new compounds with antimalarial action, capable of acting in the various stages of development of the parasite. This study aims to evaluate the in vitro antimalarial activity of P. falciparum from 18 new compounds of the quinazolinone alkaloid subclass, calculating the IC50 value. The compounds were synthesized in the Laboratory of Organic and Pharmaceutical Chemistry of the Department of Chemical Sciences, Faculty of Pharmacy of the University of Porto by the group of Doctor Teacher Maria Emília Sousa. For the 3 compounds with the best biological activity SL4C, SL5C and SL20C, the cytotoxicity was evaluated in mammalian cells V79 through the MTT assay and the Selectivity Index (IS) was calculated, haemotoxicity (hemolytic activity) and the ability to inhibit hemozoin (β-hematin) polymerization in vitro. The alkaloid compounds of quinazolinone SL4C, SL5C and SL20C demonstrated efficacy in inhibiting the growth of P. falciparum in vitro with IC50 ≤ 0,2 μM. These compounds did not present hemolytic activity in vitro. Inhibition of hemozoin (β-hematin) polymerization in vitro of the compounds at the doses tested was not detected. The compounds were not found to be cytotoxic in vitro in non-tumor mammalian cells V79 (LD50 ≤ 14 μM), with an IS value ≥ 19. The quinazolinones SL4C, SL5C and SL20C have characteristics with potential for the development as new antimalarial drugs

Does Vitamin D Status Influence the Exertional Stress Induced Neutrophil Response to 2 Hours Running at 70% ˙V O2max?

Objective.—It is well established that endurance exercise depresses neutrophil function, a key immune response in post-exercise recovery. Considering vitamin D (25(OH)D) role in immune maintenance, the study aimed to determine whether circulating 25(OH)D concentration influences bacterially challenged neutrophil degranulation in responses to endurance running. Methods.—Male runners (n = 8) completed two hydration interventions (euhydrated and dehydrated) in a randomised order during 2 h running at 70% ˙V O2max in thermoneutral conditions (0.4 and 3.1%bodymass loss, 290 and 303 mOsmol·kg−1 plasma osmolality, and −0.9 and −5.8% plasma volume change, respectively). Immediately and 1 h post-exercise participants received water (equating to × 1.5 body mass loss). Venous blood samples were collected pre- and post-exercise, and during recovery. Whole blood was used to determine total circulatory neutrophil counts and theirmaturity status. Additionally, 1000 μl of whole blood was incubated with 1 μg·mL−1 of lipopolysaccharide for 1 h at 37ºC. Aliquots were assayed for plasma elastase by ELISA. Serum and plasma samples were also analysed for 25(OH)D and IL-8 concentrations, respectively. Results.—Sub-optimal 25(OH)D levels were observed in three runners (50.1 ± 14.7 nmol·L−1), while five runners presented optimal 25(OH)D levels (99.1 ± 18.6 nmol·L−1) (P = .008), with only one athlete having insufficient levels (<50 nmol·L−1). A time effect (pre- to post-exercise increase) was observed for neutrophil degranulation (P = .001) and IL-8 response (P = .003), with no differences between the groups.A significant negative correlation was observed between serum 25(OH)D concentration and IL-8 responses in the euhydrated trial during recovery (P = .031). Conclusions.—The current study suggests that circulating 25(OH)D concentration has little impact on IL-8 or bacterially challenged neutrophil functional responses to endurance running. It is likely that the levels of 25(OH)D within the cohort were not low enough to induce substantial immune perturbations. However, the negative correlation between 25 (OH)D status and IL-8 responses in the euhydrated state warrants further investigation

Does Vitamin D Status Influence the Exertional Stress Induced Neutrophil Response to 2 Hours Running at 70% ˙V O2max?

Objective.—It is well established that endurance exercise depresses neutrophil function, a key immune response in post-exercise recovery. Considering vitamin D (25(OH)D) role in immune maintenance, the study aimed to determine whether circulating 25(OH)D concentration influences bacterially challenged neutrophil degranulation in responses to endurance running. Methods.—Male runners (n = 8) completed two hydration interventions (euhydrated and dehydrated) in a randomised order during 2 h running at 70% ˙V O2max in thermoneutral conditions (0.4 and 3.1%bodymass loss, 290 and 303 mOsmol·kg−1 plasma osmolality, and −0.9 and −5.8% plasma volume change, respectively). Immediately and 1 h post-exercise participants received water (equating to × 1.5 body mass loss). Venous blood samples were collected pre- and post-exercise, and during recovery. Whole blood was used to determine total circulatory neutrophil counts and theirmaturity status. Additionally, 1000 μl of whole blood was incubated with 1 μg·mL−1 of lipopolysaccharide for 1 h at 37ºC. Aliquots were assayed for plasma elastase by ELISA. Serum and plasma samples were also analysed for 25(OH)D and IL-8 concentrations, respectively. Results.—Sub-optimal 25(OH)D levels were observed in three runners (50.1 ± 14.7 nmol·L−1), while five runners presented optimal 25(OH)D levels (99.1 ± 18.6 nmol·L−1) (P = .008), with only one athlete having insufficient levels (<50 nmol·L−1). A time effect (pre- to post-exercise increase) was observed for neutrophil degranulation (P = .001) and IL-8 response (P = .003), with no differences between the groups.A significant negative correlation was observed between serum 25(OH)D concentration and IL-8 responses in the euhydrated trial during recovery (P = .031). Conclusions.—The current study suggests that circulating 25(OH)D concentration has little impact on IL-8 or bacterially challenged neutrophil functional responses to endurance running. It is likely that the levels of 25(OH)D within the cohort were not low enough to induce substantial immune perturbations. However, the negative correlation between 25 (OH)D status and IL-8 responses in the euhydrated state warrants further investigation.</p

Factors contributing to the mental health outcomes of carers during the transition of their family member to residential aged care : a systematic search and narrative review

Objectives: The transition of an older family member into a residential aged care facility (RACF) is often challenging for both the person being admitted and their family carer. This review aimed to identify the protective and contributing factors to adverse mental health outcomes among family carers following the decision to move a family member to a RACF. Method: A search of CINAHL, PubMed and PsycINFO was conducted for empirical papers published in English between 2004 and 2019, exploring the mental health or quality of life (QoL) of family carers of those recently admitted, or considering admission, to a RACF. Articles were reviewed by two authors for inclusion. Results: Twenty-three studies met the inclusion criteria. Pre-existing depressive symptoms and poor subjective health were related to adverse mental health outcomes following admission. Information from the facility, support to change roles, and factors related to carer’s health and demographics, were associated with changes in the mental health outcomes of carers during the transition of their relative to a RACF. Key protective factors of carer’s mental health outcomes following the transition of their relative to a RACF are flow and transparency of information between carer and the facility staff, and staff efforts to involve carers in providing emotional support to their relative, in monitoring care, and advocating for their quality of life. Conclusion: There is evidence to suggest factors such lack of flow and transparency of information between carer and the facility staff may predispose carers to poor mental health and QoL following the transition of a relative to a RACF. Key protective factors of carer’s mental health following admission are staff efforts to involve carers in providing emotional support to their relative, in monitoring care, and advocating for their quality of life. This review also indicates that the combination of factors that puts family carers more at risk of poor mental health and lower quality of life throughout the transition period. Policy and practice should follow recommendations that consider a combination of the above factors when addressing the needs of family carers before and after admission of an older person to RACF. © 2022, The Author(s)

Gut-training: The impact of two weeks repetitive gut-challenge during exercise on gastrointestinal status, glucose availability, fuel kinetics, and running performance.

Background: Due to gastrointestinal tract adaptability, the study aimed to determine the impact of two weeks gut-training protocol over two weeks on gastrointestinal status, blood glucose availability, fuel kinetics, and running performance. Methods: Endurance runners (n= 25) performed a gut-challenge trial (GC1), comprising of 2 h running exercise at 60% VO2max whilst consuming gel-discs containing 30 g carbohydrates (2:1 glucose-fructose, 10% w/v) every 20 min, and a 1 h distance test. Participants were then randomly assigned to a carbohydrate gel-disc (CHO-S), carbohydrate food (CHO-F), or placebo (PLA) gut-training group for a two weeks repetitive gut-challenge intervention. Participants then repeated a second gut-challenge trial (GC2). Results: Gastrointestinal symptoms reduced in GC2 on CHO-S (60%; p= 0.008) and CHO-F (63%; p= 0.046); reductions were greater than PLA (pThe accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author