The impact of daylight‐saving time (DST) on patients with delayed sleep‐wake phase disorder (DSWPD)

Due to time zones, sun time and local time rarely match. The difference between local and sun time, which we designate by Solar Jet Lag (SoJL), depends on location within a time zone and can range from zero to several hours. Daylight Saving Time (DST) simply adds one hour to SoJL, independent of location. We hypothesized that the impact of DST, is particularly problematic in patients with Delayed Sleep-Wake Phase Disorder (DSWPD), worsening their sleep debt. DSWPD is characterized by a chronic misalignment between the internal and social timing, reflected by an inability to fall asleep and wakeup at conventional or socially acceptable times. We analysed the clinical records of 162 DSWPD patients from a sleep medicine centre in Lisbon, Portugal (GMTzone), and separated them into two groups: the ones diagnosed across DST or Standard Time (ST). We included 82 patients (54.9% male; age: median [Q1, Q3] 34.5 [25.0, 45.3]; range 16-92; 54 in DST and 28 in ST) who had Dim Light Melatonin Onset (DLMO) measured as a marker for the circadian phase and sleep timing (onset, SO, mid-point, MS and end, SE) self-reported separately for work- and work-free days. Differences between ST and DST were compared using Mann-Whitney or Student's t tests. On a weekly average, patients in DST slept an hour less (62 min. pinfo:eu-repo/semantics/acceptedVersioninfo:eu-repo/semantics/publishedVersio

Integrated utilization of grape skins from white grape pomaces

An approach for the integrated utilization of grape skins from white grape pomaces has been proposed. This consists of consecutive or simultaneous extraction of grape skins with neutral organic solvent and water under reflux. Organic extract is a valuable raw material for the isolation of oleanolic acid. The aqueous extract (ca 50%, w/w) is composed of essentially hexoses and suitable for the high yield (till 51%) bioethanol production at a maximum specific cell growth rate (μmax) of 0.29 h−1. The remained extracted grape skins are the complex of structural polysaccharides embedded into cutinous matrix. Extracted grape skins were shown to be a prospective raw material for the production of low-density boards (d ≤ 0.40) for insulation needs. The boards produced from grape skins and bind of 8% urea–formaldehyde resin revealed reasonable tensile strength (0.4 MPa) and a low thermal conductivity (0.09–0.12 W (m K)−1) over a wide range of temperatures (40–200 °C).info:eu-repo/semantics/publishedVersio

Validation of the Portuguese Variant of the Munich Chronotype Questionnaire (MCTQPT)

This work was supported by the Fundação para a Ciência e Tecnologia (FCT) Ph.D. research Grants PDE/BDE/114584/2016 and PDE/BDE/127787/2016 and the Bial Foundation (135/18).Introduction: Differences in the manner circadian clocks entrain to the 24-h day are expressions of different chronotypes that can range from extreme early to extreme late, from proverbial larks to owls. The Morningness Eveningness Questionnaire (MEQ) was one of the first to assess daily preference based on subjective self-assessment – a psychological construct. The later developed Munich Chronotype Questionnaire (MCTQ) uses instead the actual sleep timing to assess chronotype. It calculates the mid-sleep point, halfway between onset and offset on work-free days (MSF), which is then corrected for potential oversleep on free days compensating for sleep debt accumulated over the workweek (MSFsc). MSFsc is expressed in local time and is thought to be a proxy for “phase of entrainment” of the circadian clock. The MCTQ-derived chronotype is therefore a biological construct. In the present report, we validate the Portuguese variant (MCTQPT) of the MCTQ. Portugal is of particular interest, since it is thought to consist of especially late chronotypes. Methods: We have used three methods to assess the timing of daily behavior, namely, the chronotype (MCTQ), the daily preference (rMEQ), and a simple self-assessment (time-of-day type). A total of 80 healthy adults living in Portugal, with age and sex distributed according to the Portuguese population, were recruited. We analyzed 4 weeks of continuous records of actimetry data to validate the MCTQPT and used the rMEQ to compare between a biological chronotype (sleep timing) and a psychological chronotype (daily preference). MCTQ variables were analyzed by descriptive statistics; correspondence between measurements was done by Spearman correlations or cross-tabulation; in a subset of 41 individuals, test–retest reliability was assessed. Results: MCTQ-derived variables (MSF, MSW, MSFsc) correlated highly with their counterparts calculated from actimetry (MSW: rho = 0.697; MSF: rho = 0.747; MSFsc: rho = 0.646; all p < 0.001). The MCTQ assessment of the chronotype showed good test–retest reliability (rho = 0.905; p < 0.001). The rMEQ score correlates with MSFsc (rho = −0.695; p < 0.001), and the agreement for the self-assessment with the MSFsc was fair (kw = 0.386; p < 0.001). Conclusion: The Portuguese variant of the MCTQ revealed to be a reliable questionnaire to assess the chronotype for the Portuguese adult population, as previously reported for other countries.publishersversionpublishe

Structural characterization of lignin from Grape Stalks (Vitis vinifera L.)

The chemical structure of lignin from grape stalks, an abundant waste of winemaking, has been studied. The dioxane lignin was isolated from extractive- and protein-free grape stalks (Vitis vinifera L.) by modified acidolytic procedure and submitted to a structural analysis by wet chemistry (nitrobenzene and permanganate oxidation (PO)) and spectroscopic techniques. The results obtained suggest that grape stalk lignin is an HGS type with molar proportions of p-hydroxyphenyl (H), guaiacyl (G) and syringyl (S) units of 3:71:26. Structural analysis by 1H and 13C NMR spectroscopy and PO indicates the predominance of β-O-4′ structures (39% mol) in grape stalk lignin together with moderate amounts of β-5′, β-β, β-1′, 5-5′, and 4-O-5′ structures. NMR studies also revealed that grape lignin should be structurally associated with tannins. The condensation degree of grape stalks lignin is higher than that of conventional wood lignins and lignins from other agricultural residues.N/

Structural characterization of polysaccharides isolated from grape stalks of Vitis vinifera L.

The main structural polysaccharides of grape stalks are cellulose, heteroxylan, and glucan. Cellulose contributes 30.3% of grape stalk matter and has an unusually high degree of the crystallinity (75.4%). Among hemicelluloses, xylan was the most abundant one, contributing ∼12% to the weight. The heteroxylan was isolated from the corresponding peracetic holocellulose by DMSO extraction followed by precipitation in ethanol. The Mw of heteroxylan (19.0kDa) was assessed by size exclusion chromatography (SEC) and the structure was inferred by methanolysis and methylation linkage analysis, as well as 1D and 2D nuclear magnetic resonance (NMR) spectroscopy. The heteroxylan is a partially acetylated (DS=0.49) glucuronoxylan possessing the main backbone composed by β-(1→4)-linked d-xylopyranosyl units ramified with α-(1→2)-linked 4-O-methyl-α-d-glucuronosyl residues (MeGlcpA) at a molar ratio 25:1. The isolated heteroxylan contained concomitant β-glucan (ca. 15%), whose structure was elucidated by methylation linkage analysis and by NMR spectroscopy. The results obtained revealed mixed β-(1→3; 1→4)-d-glucan with a molar ratio of β-(1→3)- to β-(1→4)-linked glucopyranosyl units of 1:2.info:eu-repo/semantics/publishedVersio

Methylene blue associated with maghemite nanoparticles has antitumor activity in breast and ovarian carcinoma cell lines

Background: Cancer constitutes group of diseases responsible for the second largest cause of global death, and it is currently considered one of the main public health concerns nowadays. Early diagnosis associated with the best choice of therapeutic strategy, is essential to achieve success in cancer treatment. In women, breast cancer is the second most common type, whereas ovarian cancer has the highest lethality when compared to other neoplasms of the female genital system. The present work, therefore, proposes the association of methylene blue with citrate-coated maghemite nanoparticles (MAGCIT–MB) as a nanocomplex for the treatment of breast and ovarian cancer. Results: In vitro studies showed that T-47D and A2780 cancer cell lines underwent a significant reduction in cell viability after treatment with MAGCIT–MB, an event not observed in non-tumor (HNTMC and HUVEC) cells and MDA-MB-231, a triple-negative breast cancer cell line. Flow cytometry experiments suggest that the main mechanism of endocytosis involved in the interiorization of MAGCIT–MB is the clathrin pathway, whereas both late apoptosis and necrosis are the main types of cell death caused by the nanocomplex. Scanning electron microscopy and light microscopy reveal significant changes in the cell morphology. Quantification of reactive oxygen species confirmed the MAGCIT–MB cytotoxic mechanism and its importance for the treatment of tumor cells. The lower cytotoxicity of individual solution of maghemite nanoparticles with citrate (MAGCIT) and free methylene blue (MB) shows that their association in the nanocomplex is responsible for its enhanced therapeutic potential in the treatment of breast and ovarian cancer in vitro. Conclusions: Treatment with MAGCIT–MB induces the death of cancer cells but not normal cells. These results highlight the importance of the maghemite core for drug delivery and for increasing methylene blue activity, aiming at the treatment of breast and ovarian cancer

Exploring the correlations between epi indicators of COVID-19 and the concentration of pharmaceutical compounds in Wastewater Treatment Plants in Northern Portugal

The COVID-19 pandemic caused by the SARS-CoV-2 virus led to changes in the lifestyle and human behaviour, which resulted in different consumption patterns of some classes of pharmaceuticals including curative, symptom-relieving, and psychotropic drugs. The trends in the consumption of these compounds are related to their concentrations in wastewater systems, since incompletely metabolised drugs (or their metabolites back transformed into the parental form) may be detected and quantified by analytical methods. Pharmaceuticals are highly recalcitrant compounds and conventional activated sludge processes implemented in wastewater treatment plants (WWTP) are ineffective at degrading these substances. As a results, these compounds end up in waterways or accumulate in the sludge, being a serious concern given their potential effects on ecosystems and public health. Therefore, it is crucial to evaluate the presence of pharmaceuticals in water and sludge to assist in the search for more effective processes. In this work, eight pharmaceuticals from five therapeutic classes were analysed in wastewater and sludge samples collected in two WWTP located in the Northern Portugal, during the third COVID-19 epidemic wave in Portugal. The two WWTP demonstrated a similar pattern with respect to the concentration levels in that period. However, the drugs loads reaching each WWTP were dissimilar when normalising the concentrations to the inlet flow rate. Acetaminophen (ACET) was the compound detected at highest concentrations in aqueous samples of both WWTP (98. 516 g L1 in WWTP2 and 123. 506 g L1in WWTP1), indicating that this drug is extensively used without the need of a prescription, known of general public knowledge as an antipyretic and analgesic agent to treat pain and fever. The concentrations determined in the sludge samples were below 1.65 µg g1 in both WWTP, the highest value being found for azithromycin (AZT). This result may be justified by the physico-chemical characteristics of the compound that favour its adsorption to the sludge surface through ionic interactions. It was not possible to establish a clear relationship between the incidence of COVID-19 cases in the sewer catchment and the concentration of drugs detected in the same period. However, looking at the data obtained, the high incidence of COVID-19 in January 2021 is in line with the high concentration of drugs detected in the aqueous and sludge samples but prediction of drug load from viral load data was unfeasible.This study was supported by the Competitiveness and Internationalisation Operational Programme, Lisbon Regional Operational Programme and Algarve Regional Operational Programme with the support of FEDER, through the Incentive Scheme: research and development activities and investment in testing and optimisation (upscaling) infrastructures in the context of COVID-19, through the Project “SARS CONTROL: Evaluation of the impacts of SARS-CoV-2 on the urban water cycle and the downstream effects on Public Health" (Ref. 070076). Acknowledge is also due to the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit, and by LABBELS – Associate Laboratory in Biotechnology, Bioengineering and Microelectromechanical Systems, LA/P/0029/2020. Strategic funding from FCT to cE3c and BioISI Research Units (UIDB/00329/2020 and UIDB/04046/2020) and to the Associate Laboratory CHANGE (LA/P/0121/2020) is also gratefully acknowledged. ARS holds an FCT grant SFRH/BD/131905/2017 and COVID/BD/151951/2021.ARLR and MFRP acknowledge the financial support from LA/P/0045/2020 (ALiCE), UIDB/50020/2020 and UIDP/50020/2020 (LSRE-LCM), funded by national funds through FCT/MCTES (PIDDAC). ARLR acknowledges FCT funding under DL57/2016 Transitory Norm Programme.info:eu-repo/semantics/publishedVersio

Differentiation of Bacillus pumilus and Bacillus safensis using MALDI-TOF-MS

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) despite being increasingly used as a method for microbial identification, still present limitations in which concerns the differentiation of closely related species. Bacillus pumillus and Bacillus safensis, are species of biotechnological and pharmaceutical significance, difficult to differentiate by conventional methodologies. In this study, using a well-characterized collection of B. pumillus and B. safensis isolates, we demonstrated the suitability of MALDI-TOF-MS combined with chemometrics to accurately and rapidly identify them. Moreover, characteristic species-specific ion masses were tentatively assigned, using UniProtKB/Swiss-Prot and UniProtKB/TrEMBL databases and primary literature. Delineation of B. pumilus (ions at m/z 5271 and 6122) and B. safensis (ions at m/z 5288, 5568 and 6413) species were supported by a congruent characteristic protein pattern. Moreover, using a chemometric approach, the score plot created by partial least square discriminant analysis (PLSDA) of mass spectra demonstrated the presence of two individualized clusters, each one enclosing isolates belonging to a species-specific spectral group. The generated pool of species-specific proteins comprised mostly ribosomal and SASPs proteins. Therefore, in B. pumilus the specific ion at m/z 5271 was associated with a small acid-soluble spore protein (SASP O) or with 50S protein L35, whereas in B. safensis specific ions at m/z 5288 and 5568 were associated with SASP J and P, respectively, and an ion at m/z 6413 with 50S protein L32. Thus, the resulting unique protein profile combined with chemometric analysis, proved to be valuable tools for B. pumilus and B. safensis discrimination, allowing their reliable, reproducible and rapid identification.Dr. Kasthuri Venkateswaran, Dr. Irene Ouoba, Dr. Joseph W. Kloepper, Dr. Cecilie From and Dr. Maria Morea are gratefully acknowledged for providing isolates FO-36bT, SAFN-027, SAFN-037, KL-052, 51-3C and 82-2C; Bs31; SE 49 (AP3) and SE 52 (AP7); FEL 55, UNG22 and MIL46, respectively. Raquel Branquinho was supported by a PhD fellowship (Ref. SFRH/BD/61410/2009) and Clara Sousa by a post-doctoral fellowship (Ref. SFRH/BPD/70548/2010), from FCT (Fundacao para a Ciencia e Tecnologia, Portugal). Hugo Osorio acknowledges the funding from QREN-FEDER through the Operational Program ON. 2 - O Novo Norte. IPATIMUP is an Associate Laboratory of the Portuguese Ministry of Science, Technology and Higher Education and is partially supported by the Portuguese Foundation for Science and Technology. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competin