ARAP3 protects from excessive formylated peptide-induced microvascular leakage by acting on endothelial cells and neutrophils.

Vascular permeability is temporarily heightened during inflammation, but excessive inflammation-associated microvascular leakage can be detrimental, as evidenced in the inflamed lung. Formylated peptides regulate vascular leakage indirectly via formylated peptide receptor-1 (FPR1)-mediated recruitment and activation of neutrophils. Here we identify how the GTPase-activating protein ARAP3 protects against formylated peptide-induced microvascular permeability via endothelial cells and neutrophils. In vitro, Arap3-/- endothelial monolayers were characterised by enhanced formylated peptide-induced permeability due to upregulated endothelial FPR1 and enhanced vascular endothelial cadherin internalisation. In vivo, enhanced inflammation-associated microvascular leakage was observed in Arap3-/- mice. Leakage of plasma protein into the lungs of Arap3-/- mice increased within hours of formylated peptide administration. Adoptive transfer experiments indicated this was dependent upon ARAP3 deficiency in both immune and non-immune cells. Bronchoalveolar lavages of formylated peptide-challenged Arap3-/- mice contained neutrophil extracellular traps (NETs). Pharmacological inhibition of NET formation abrogated excessive microvascular leakage, indicating a critical function of NETs in this context. The observation that Arap3-/- mice developed more severe influenza suggests these findings are pertinent to pathological situations characterised by abundant formylated peptides. © 2024 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Eco-friendly management of harmful cyanobacterial blooms in eutrophic lakes through vertical flow multi-soil-layering technology.

Eutrophication has led to the widespread occurrence of cyanobacterial blooms. Toxic cyanobacterial blooms with high concentrations of microcystins (MCs) have been identified in the Lalla Takerkoust reservoir in Morocco. The objective of this study was to evaluate the efficiency of the Multi-Soil-Layering (MSL) ecotechnology in removing natural cyanobacterial blooms from the lake. Two MSL pilots were used in rectangular glass tanks (60 × 10 × 70 cm). They consisted of permeable layers (PLs) made of pozzolan and a soil mixture layer (SML) containing local soil, ferrous metal, charcoal and sawdust. The main difference between the two systems was the type of local soil used: sandy soil for MSL1 and clayey soil for MSL2. Both MSL pilots effectively reduced cyanobacterial cell concentrations in the treated water to very low levels (0.09 and 0.001 cells/mL). MSL1 showed a gradual improvement in MC removal from 52 % to 99 %, while MSL2 started higher at 90 % but dropped to 54% before reaching 86%. Both MSL systems significantly reduced organic matter levels (97.2 % for MSL1 and 95.8 % for MSL2). Both MSLs were shown to be effective in removing cyanobacteria, MCs, and organic matter with comparable performance.

Identification and structural characterization of a novel (+)-catechin-caffeic acid adduct present in wines.

A new compound with a molecular ion mass of m/z 467 in the negative ion mode was found to occur in a white wine aged 30 months in bottle. In this latter, fragment ions compatible with the loss of a carboxylic acid (-44 a.m.u.), a caffeic acid unit (-178 a.m.u.), and a Retro-Diels Alder (-152 a.m.u.) were observed. The present work reports the synthesis of a (+)-catechin-caffeic acid adduct resulting from the condensation reaction between caffeic acid and (+)-catechin. The structural characterization by NMR showed that this adduct is formed by the linkage between carbon 8 at ring A from (+)-catechin and carbon 9 from caffeic acid. In addition, the similarity in the HPLC retention time and UV-Visible spectra of the synthesized compound with the one detected in white wine and the bottling experiments, confirms the presence of this novel (+)-catechin-derived compound in those matrices.

Tracing the fate of microcystins from irrigation water to food chains: Studies with Fragaria vulgaris and Meriones shawi.

Microcystins (MCs) are cyanobacterial toxins that can negatively impact human and animal health. This study investigated the bioaccumulation, transfer, depuration, and health risks of MCs in strawberry plants (Fragaria vulgaris) and Meriones shawi animals. The plants were irrigated with 1, 5, 10, and 20 µg/L MCs for 60 days (bioaccumulation phase) and then with clean water for 30 days (depuration phase). The harvested plants (roots and leaves) were then prepared in an aliquot form and used as feed for Meriones shawi. Liquid chromatography-mass spectrometry (LC/MS/MS) was used to measure MC concentrations in plant and animal tissues. The bioaccumulation of MCs was found to be highest in the roots, followed by leaves, fruits, liver, stomach, and fecal matter. The bioaccumulation factor (BAF) was highest in perlite (8.48), followed by roots (5.01), leaves (1.55), stomach (0.87), and fecal matter (1.18), indicating that the parts with high bioaccumulation factor had high translocation of MCs. The transfer of MCs to animal organs was low, and the daily toxin intake of adult consumers of strawberry fruit irrigated with 1, 5, 10, and 20 µg/L MC did not exceed the WHO-recommended limit of 0.04 µg MC-LR/Kg of bw/day. However, fruits from plants irrigated with 10 and 20 µg/L may pose a moderate health risk to children (25 Kg bw), and Meriones' consumption of leaves may pose a significant health risk. After the depuration phase, MC concentration in perlite, roots, leaves, and fruits decreased, indicating that depuration reduced the danger of MC transmission and bioaccumulation. The study also found that glutathione reductase and glutathione S-transferase activity were essential in the depuration of MCs in the tested plants. The findings suggest that legislation regulating the quality of irrigation water in terms of MC concentrations is necessary to prevent detrimental consequences to crops and human exposure.

Bioprospecting for industrially relevant exopolysaccharide-producing cyanobacteria under Portuguese simulated climate.

Cyanobacterial exopolysaccharides (EPS) are potential candidates for the production of sustainable biopolymers. Although the bioactive and physicochemical properties of cyanobacterial-based EPS are attractive, their commercial exploitation is limited by the high production costs. Bioprospecting and characterizing novel EPS-producing strains for industrially relevant conditions is key to facilitate their implementation in various biotechnological applications and fields. In the present work, we selected twenty-five Portuguese cyanobacterial strains from a diverse taxonomic range (including some genera studied for the first time) to be grown in diel light and temperature, simulating the Portuguese climate conditions, and evaluated their growth performance and proximal composition of macronutrients. Synechocystis and Cyanobium genera, from marine and freshwater origin, were highlighted as fast-growing (0.1-0.2 g L-1 day-1) with distinct biomass composition. Synechocystis sp. LEGE 07367 and Chroococcales cyanobacterium LEGE 19970, showed a production of 0.3 and 0.4 g L-1 of released polysaccharides (RPS). These were found to be glucan-based polymers with high molecular weight and a low number of monosaccharides than usually reported for cyanobacterial EPS. In addition, the absence of known cyanotoxins in these two RPS producers was also confirmed. This work provides the initial steps for the development of cyanobacterial EPS bioprocesses under the Portuguese climate.

Combined Neonatal Alloimmune Neutropenia and Thrombocytopenia in Dizygotic Twins Conceived After Oocyte Donation.

Neonatal alloimmune thrombocytopenia (NAIT) and neonatal alloimmune neutropenia (NAIN) may have severe consequences in the neonatal period. We report two dizygotic twins conceived after donated oocytes, suffering NAIT and NAIN in the context of alloantibodies to human platelet antigens (anti-HPA-5b) and human leukocyte antigens (anti-HLA class I). Genotyping demonstrated paternal homozygosity for HPA-5a, while the neonates were heterozygous for HPA-5b.

Reduced G protein signaling despite impaired internalization and ß-arrestin recruitment in patients carrying a CXCR4Leu317fsX3 mutation causing WHIM syndrome.

WHIM syndrome is an inherited immune disorder caused by an autosomal dominant heterozygous mutation in CXCR4. The disease is characterized by neutropenia/leukopenia (secondary to retention of mature neutrophils in bone marrow), recurrent bacterial infections, treatment-refractory warts, and hypogammaglobulinemia. All mutations reported in WHIM patients lead to the truncations in the C-terminal domain of CXCR4, R334X being the most frequent. This defect prevents receptor internalization and enhances both calcium mobilization and ERK phosphorylation, resulting in increased chemotaxis in response to the unique ligand CXCL12. Here, we describe 3 patients presenting neutropenia and myelokathexis, but normal lymphocyte count and immunoglobulin levels, carrying what we believe to be a novel Leu317fsX3 mutation in CXCR4, leading to a complete truncation of its intracellular tail. The analysis of the L317fsX3 mutation in cells derived from patients and in vitro cellular models reveals unique signaling features in comparison with R334X mutation. The L317fsX3 mutation impairs CXCR4 downregulation and ß-arrestin recruitment in response to CXCL12 and reduces other signaling events - including ERK1/2 phosphorylation, calcium mobilization, and chemotaxis - all processes that are typically enhanced in cells carrying the R334X mutation. Our findings suggest that, overall, the L317fsX3 mutation may be causative of a form of WHIM syndrome not associated with an augmented CXCR4 response to CXCL12.

Using Social Media Categorical Reactions as a Gateway to Identify Hate Speech in COVID-19 News.

The spread of COVID-19 news on social media provided a particularly prolific ground for emotional commotion, disinformation and hate speech, as uncertainty and fear grew by the day. In this paper, we examine the media coverage of the COVID-19 outburst in Portugal (March-May 2020), the subsequent emotional engagement of audiences and the entropy-based emotional controversy generated as a gateway to detect the presence of hate speech, using computer-assisted qualitative data analysis (CAQDAS) embedded in a cross-sectional descriptive methodology. Our results reveal that negative and volatile categorical emotions ("Angry", "Haha" and "Wow") serve as main engines for controversy, and that controversial news have the highest sharing ratio. Moreover, using a small sample of the most controversial news with the highest overall emotional engagement, we establish a relation between the entropy-based emotional controversy obtained from Facebook's click-based reactions and the presence of cultural and ethnic hate speech, plausibly confirming the click-based categorical emotions as a gateway to hatred comment pools. In doing so, we also reveal that negative emotions alone do not always indicate the presence of hate speech, which may sprout in seemingly humorous social media posts where irony proliferates, and negativity is not apparent. This work adds to the literature on social media categorical emotion detection and its implications for the detection of hate speech.

Cork, a Natural Choice to Wine?

This review presents the most recent data on the state-of-the-art of the main compounds present in cork, their interaction with wine, and the impact that natural stoppers may have on wines' physical-chemical and sensory properties. According to the recent scientific literature, the chemical composition of cork and the scientific relevance of the compounds extract from cork to wine over time are reviewed. Furthermore, the effect of cork compounds transfer into wines during post-bottling is also discussed, as well as their impact on the organoleptic (colour and taste) of wines. This knowledge is essential for the decision-making process undertaken by wine producers to select the stopper most suitable for their wines. In addition, sustainability is also a topic addressed since it is a natural product that generates some waste as well as the way in which this industry is adapting to the closure of the waste cycle.

The Impact of Storage Conditions and Bottle Orientation on the Evolution of Phenolic and Volatile Compounds of Vintage Port Wine.

This work evaluates the influence of the cellar conditions and bottle orientation, on the phenolic and volatile composition of a Vintage Port wine, sealed with natural cork stoppers, for 44 months post-bottling. The storage was performed in two different cellars, namely a cellar A with controlled temperature and humidity, and a cellar B, representing a traditional cellar, with uncontrolled temperature and humidity. The impact of bottle orientation was studied in cellar A, where the bottles were stored in horizontal and vertical positions. The phenolic and volatile composition of the bottled Vintage Port wine were analyzed after 6, 15 and 44 months. The results unveiled that the cellar conditions and bottle orientation had an impact in Port wine composition which was higher at 44 months post-bottling. The samples stored in the traditional cellar unveiled significantly higher yellow tones, lower tannin specific activity, and higher levels of furfural and 5-methylfurfural. Furthermore, the samples stored in the horizontal position revealed significant higher levels of total proanthocyanidins and higher tannin specific activity than the samples stored in the vertical position. Interestingly, for the first time to our knowledge, an ellagitannin-derived compound (Corklin) was detected in Vintage Port wines stored in the horizontal position, which results from the reaction of cork constituents with phenolic compounds present in wines.

The effectiveness of Sustained Natural Apophyseal Glide on Flexion Rotation Test, pain intensity, and functionality in subjects with Cervicogenic Headache: A Systematic Review of Randomized Trials.

OBJECTIVE: To determine the effect of sustained natural apophyseal glide (SNAG) on Flexion Rotation Test, pain intensity, and functionality in subjects with Cervicogenic Headache (CH). METHODS: The research was conducted on five computerized databases PubMed/Medline, Web of Science, PEDro, Lilacs, and Cochrane Library (CENTRAL), using the keywords combination: (sustained natural apophyseal glide OR SNAG OR joint mobilization OR Mulligan) AND (cervicogenic headache) according to PRISMA guidelines. The methodological quality of the included studies was analyzed using the Physiotherapy Evidence Database (PEDro) scale. RESULTS: Eight articles fulfilled the eligibility criteria and were included in the review. The selected studies had a methodological quality of 6.6/10 on the PEDro scale and included a total of 357 participants. The SNAG significantly improved pain, Flexion Rotation Test and reduced functional symptoms. CONCLUSION: The available evidence suggests that SNAG may be a relevant intervention for CH.

Morphological, molecular, and biochemical study of cyanobacteria from a eutrophic Algerian reservoir (Cheffia).

The cyanobacteria management in water bodies requires a deep knowledge of the community composition. Considering the reliable and thorough information provided by the polyphasic approach in cyanobacteria taxonomy, here we assess the cyanobacterial community structure of the Cheffia reservoir from Algeria. Cyanobacteria were identified on the basis of morphological traits and next-generation sequencing (NGS); toxins-related genes were localized in addition to the identification of toxins; temperature and nutrient level of water samples were also determined. The polyphasic approach was essential for cyanobacteria investigation; 28 genera were identified through 16S rRNA metabarcoding with the dominance of taxa from Microcystis (34.2%), Aphanizomenon (20.1%), and Planktothrix (20.0%), and morphological analysis revealed the association in this water body of five species within the genus Microcystis: M. aeruginosa, M. novacekii, M. panniformis, M. ichthyoblabe, and M. flos-aquae. The presence of mcyE genotypes was detected; moreover, HPLC-PDA and LC-ESI-MS/MS revealed the production of microcystin-LR. Results obtained in our study are very important since this ecosystem is used for water supply and irrigation; as a consequence, a good water management plan is essential.

Interaction between salivary proteins and cork phenolic compounds able to migrate to wine model solutions.

This work reports the study of the interaction of human salivary proteins (SP) with phenolic compounds that migrate from cork stoppers to wine. This study yields valuable data to understand the influence that these compounds may have on the sensory perception of wine from an astringency perspective. For that, three cork fractions containing the phenolic compounds that migrate in greater amounts from cork to model wine solutions were selected. Fraction M1 contains gallic acid, protocatechuic acid, vanillin and protocatechuic aldehyde; fraction M2 comprises essentially gallic acid and ellagic acid, as well as castalagin and dehydrocastalagin; and fraction M3 contains the two isomeric ellagitannins castalagin and vescalagin. The reactivity of each fraction towards SP was M3 > M2 > M1. Within M3 fraction, castalagin showed a higher ability to precipitate SP (mainly aPRPs, statherin and P-B peptide) comparatively to vescalagin. In M1 fraction, caffeic and sinapic acids were the compounds with the highest interaction with SP, mainly cystatins. In addition, there also seems to be a matrix effect (presence of other compounds) that could be affecting these interactions.

Cyanotoxin Screening in BACA Culture Collection: Identification of New Cylindrospermopsin Producing Cyanobacteria.

Microcystins (MCs), Saxitoxins (STXs), and Cylindrospermopsins (CYNs) are some of the more well-known cyanotoxins. Taking into consideration the impacts of cyanotoxins, many studies have focused on the identification of unknown cyanotoxin(s)-producing strains. This study aimed to screen strains from the Azorean Bank of Algae and Cyanobacteria (BACA) for MCs, STX, and CYN production. A total of 157 strains were searched for mcy, sxt, and cyr producing genes by PCR, toxin identification by ESI-LC-MS/MS, and cyanotoxin-producing strains morphological identification and confirmation by 16S rRNA phylogenetic analysis. Cyanotoxin-producing genes were amplified in 13 strains and four were confirmed as toxin producers by ESI-LC-MS/MS. As expected Aphanizomenon gracile BACA0041 was confirmed as an STX producer, with amplification of genes sxtA, sxtG, sxtH, and sxtI, and Microcystis aeruginosa BACA0148 as an MC-LR producer, with amplification of genes mcyC, mcyD, mcyE, and mcyG. Two nostocalean strains, BACA0025 and BACA0031, were positive for both cyrB and cyrC genes and ESI-LC-MS/MS confirmed CYN production. Although these strains morphologically resemble Sphaerospermopsis, the 16S rRNA phylogenetic analysis reveals that they probably belong to a new genus.

Depletion of Arg/Abl2 improves endothelial cell adhesion and prevents vascular leak during inflammation.

Endothelial barrier disruption and vascular leak importantly contribute to organ dysfunction and mortality during inflammatory conditions like sepsis and acute respiratory distress syndrome. We identified the kinase Arg/Abl2 as a mediator of endothelial barrier disruption, but the role of Arg in endothelial monolayer regulation and its relevance in vivo remain poorly understood. Here we show that depletion of Arg in endothelial cells results in the activation of both RhoA and Rac1, increased cell spreading and elongation, redistribution of integrin-dependent cell-matrix adhesions to the cell periphery, and improved adhesion to the extracellular matrix. We further show that Arg is activated in the endothelium during inflammation, both in murine lungs exposed to barrier-disruptive agents, and in pulmonary microvessels of septic patients. Importantly, Arg-depleted endothelial cells were less sensitive to barrier-disruptive agents. Despite the formation of F-actin stress fibers and myosin light chain phosphorylation, Arg depletion diminished adherens junction disruption and intercellular gap formation, by reducing the disassembly of cell-matrix adhesions and cell retraction. In vivo, genetic deletion of Arg diminished vascular leak in the skin and lungs, in the presence of a normal immune response. Together, our data indicate that Arg is a central and non-redundant regulator of endothelial barrier integrity, which contributes to cell retraction and gap formation by increasing the dynamics of adherens junctions and cell-matrix adhesions in a Rho GTPase-dependent fashion. Therapeutic inhibition of Arg may provide a suitable strategy for the treatment of a variety of clinical conditions characterized by vascular leak.

On the Limits of Anthocyanins Co-Pigmentation Models and Respective Equations.

Anthocyanins co-pigmentation models with application on 1:1 complexes were revisited, and their limitations were critically commented. The flavylium multistate of species is dramatically simplified to a single acid-base equilibrium between flavylium cation and its conjugated base CB, equal to the sum of quinoidal base, hemiketal, and cis and trans-chalcones. Bearing this, a new equation that simultaneously allows calculation of the co-pigmentation constant with flavylium cation (KAH+CP) and with its conjugated base CB (KCBCP) was deduced. This equation can be used at a fixed co-pigment concentration with pH as a variable or at fixed pH and co-pigment concentration variable. A global fitting of all data allows us to calculate both association constants with good accuracy. The model was applied to the co-pigmentation of malvidin-3-glucoside with caffeine and pentagalloyl glucose (PGG). Caffeine gives rise to complexes not only with flavylium cation KAH+CP = 125 ± 7 M-1 but also with CB with KCBCP = 23 ± 3 M-1. PGG complexes exclusively with flavylium cation, KAH+CP = 914 ± 10 M-1, and the possible interaction with quinoidal base is lower than the detection limits that the inherent experimental error permits.

Predicting musculoskeletal symptoms in workers of a manufacturing company.

Background. Musculoskeletal symptoms are the leading factor responsible for sickness absence. Factors associated with the development of musculoskeletal symptoms must be identified and addressed. Objective. To investigate the 7-day prevalence of musculoskeletal symptoms by body regions, and assess the contribution of individual, lifestyle and occupational risk factors to the development of musculoskeletal disorders. Methods. The sample comprised 202 white-collar and blue-collar workers. Musculoskeletal symptoms were assessed with the Nordic questionnaire, and physical activity level and sitting time with the international physical activity questionnaire. Statistical analyses were conducted to assess the associations between individual, lifestyle and occupational category factors and musculoskeletal symptoms. Results. The 7-day prevalence of work-related musculoskeletal symptoms was 41.6% (n = 84). The highest prevalence of symptoms was reported in the lower back (18.8%), followed by the wrists/hands (17.3%), neck (15.8%), shoulder (15.3%) and ankles/feet (11.4%). A significant association was found between reporting musculoskeletal symptoms and work category. In addition, musculoskeletal symptoms were reported more often by older workers. Work category was a significant predictor of pain in the lower back and shoulder regions, and gender was a significant predictor for neck pain. Conclusions. Our results emphasize the need for worksite interventions to prevent musculoskeletal symptoms in this population.

The influence of sports practice, dominance and gender on the knee joint position sense.

BACKGROUND: Physical activity, muscle fatigue or age have been suggested as factors that positively or negatively influence the knee joint position sense (KJPS). However, conflicting results about the influence of sports practice, limb dominance and gender are found in the literature. This study aimed to assess the influence of sports practice, limb dominance and gender on the KJPS of soccer players and untrained individuals. METHODOLOGY: Sixty subjects participated in this study: 29 soccer players (SPs) and 31 untrained participants (UPs). KJPS was tested in a seated position, for the target angles of 20° and 45° of knee flexion, through an open kinetic chain technique and active repositioning method. Intergroup analysis was performed to compare KJPS between SPs and UPs and between gender, and intragroup analysis was performed to compare proprioceptive acuity between dominant and non-dominant limbs. RESULTS: Knee repositioning errors were lower in SPs than in the UPs. Repositioning errors of the dominant and non-dominant limb were not significantly different in SPs (P > 0.05), but in UPs the dominant limb showed significantly lower repositioning errors, both for 20° (P = 0.046) and 45° target (P = 0.036). There were no gender differences in the KJPS in both groups (P > 0.05). CONCLUSIONS: Proprioceptive acuity is higher in trained than in untrained participants. Dominance seems not to influence KJPS of SPs, but in UPs the dominant limb showed a higher accuracy. There were no differences in KJPS related to gender. These results suggest that sports practice, but neither gender nor limb dominance, may positively influence proprioceptive acuity.