Fluorescence imaging of lamellipodin-mediated biomolecular condensates on solid supported lipid bilayer membranes.

Biomolecular condensates play a major role in numerous cellular processes, including several that occur on the surface of lipid bilayer membranes. There is increasing evidence that cellular membrane trafficking phenomena, including the internalization of the plasma membrane through endocytosis, are mediated by multivalent protein-protein interactions that can lead to phase separation. We have recently found that proteins involved in the clathrin-independent endocytic pathway named Fast Endophilin Mediated Endocytosis can undergo liquid-liquid phase separation (LLPS) in solution and on lipid bilayer membranes. Here, the protein solution concentrations required for phase separation to be observed are significantly smaller compared to those required for phase separation in solution. LLPS is challenging to systematically characterize in cellular systems in general, and on biological membranes in particular. Model membrane approaches are more suitable for this purpose as they allow for precise control over the nature and amount of the components present in a mixture. Here we describe a method that enables the imaging of LLPS domain formation on solid supported lipid bilayers. These allow for facile imaging, provide long-term stability, and avoid clustering of vesicles and vesicle-attached features (such as buds and tethers) in the presence of multi-valent membrane interacting proteins.

The Development of a Rapid, Cost-Effective, and Green Analytical Method for Mercury Speciation.

Mercury is a naturally occurring metal found in various inorganic and organic forms within the environment. Due to its high toxicity, there is global concern regarding human exposure to this element. The combination of high-performance liquid chromatography and inductively coupled plasma mass spectrometry (HPLC-ICP-MS) is commonly used to analyze the different forms of mercury in a sample due to its high sensitivity and ability to selectively detect mercury. However, the traditional HPLC-ICP-MS methods are often criticized for their lengthy analysis times. In this study, we have refined the conventional approach by transitioning to ultra-high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry (UHPLC-ICP-MS). This modification has resulted in significant reductions in runtime as well as reagent and argon usage, thereby offering a more rapid, environmentally friendly, and cost-effective method. We successfully adapted an HPLC-ICP-MS method to UHPLC-ICP-MS, achieving the analysis of Hg2+ and MeHg+ within 1 min with a mobile phase consumption of only 0.5 mL and a sample volume of 5.0 µL; this is a major advance compared to HPLC analysis with run times generally between 5 and 10 min. The method's performance was assessed by analyzing muscle and liver tissue samples (serving as reference material) from fish, demonstrating the versatility of the method in relation to different complex matrices.

All-polysaccharide, self-healing, pH-sensitive, in situ-forming hydrogel of carboxymethyl chitosan and aldehyde-functionalized hydroxyethyl cellulose.

In situ forming hydrogels are promising for biomedical applications, especially in drug delivery. The precursor solution can be injected at the target site, where it undergoes a sol-gel transition to afford a hydrogel. In this sense, the most significant characteristic of these hydrogels is fast gelation behavior after injection. This study describes an all-polysaccharide, rapidly in situ-forming hydrogel composed of carboxymethyl chitosan (CMCHT) and hydroxyethyl cellulose functionalized with aldehyde groups (HEC-Ald). The HEC-Ald was synthesized through acetal functionalization, followed by acid deprotection. This innovative approach avoids cleavage of pyran rings, as is inherent in the periodate oxidation approach, which is the most common method currently employed for adding aldehyde groups to polysaccharides. The resulting hydrogel exhibited fast stress relaxation, self-healing properties, and pH sensitivity, which allowed it to control the release of an encapsulated model drug in response to the medium pH. Based on the collected data, the HEC-Ald/CMCHT hydrogels show promise as pH-sensitive drug carriers.

The parabasal filaments of Trichomonas vaginalis: A new filament and observations using 0.8 nm-resolution scanning electron microscopy.

Trichomonas vaginalis is the etiologic agent of trichomoniasis, the most common nonviral sexually transmitted infection worldwide, with an estimated 260 million new cases annually. T. vaginalis contains organelles common to all eukaryotic cells, uncommon cell structures such as hydrogenosomes, and a complex and elaborate cytoskeleton constituting the mastigont system. The mastigont system is mainly formed by several proteinaceous structures associated with basal bodies, the pelta-axostylar complex made of microtubules, and striated filaments named the costa and the parabasal filaments (PFs). Although the structural organization of trichomonad cytoskeletons has been analyzed using several techniques, observation using a new generation of scanning electron microscopes with a resolution exceeding 1 nm has allowed more detailed visualization of the three-dimensional organization of the mastigont system. In this study, we have investigated the cytoskeleton of T. vaginalis using a diverse range of scanning probe microscopy techniques, which were complemented by electron tomography and Fast-Fourier methods. This multi-modal approach has allowed us to characterize an unknown parabasal filament and reveal the ultrastructure of other striated fibers that have not been published before. Here, we show the differences in origin, striation pattern, size, localization, and additional details of the PFs, thus improving the knowledge of the cell biology of this parasite.

Influence of Fast Drying on the Morphology of α-Fe2O3 and FeMnO3/α-Fe2O3 Fibers Produced by Solution Blow Spinning.

α-Fe2O3 and FeMnO3/α-Fe2O3 fibers were successfully prepared via Solution Blow Spinning (SBS). The effect of drying during the SBS process on fiber morphology was evaluated by scanning electron microscopy (SEM), transmission electron microscopy (TEM), and N2 adsorption-desorption isotherms. A slow drying promoted continuous fibers with rough surfaces and lower average diameters. However, fast drying enabled the production of fibers with low densification and many surface pores with higher BET-specific surface areas. The porous fibers produced have potential applications in energy generation and storage.

Assessment and comparison of thermochemical pathways for the rice residues valorization: pyrolysis and gasification.

Lignocellulosic biomass conversion applying thermochemical routes has been postulated as an alternative for generating renewable energy. This research compares energy-driven biorefineries based on two thermochemical routes addressed to upgrade rice husk and rice straw produced in the Department of Sucre-Colombia. Initially, this research analyzes the physico-chemical and structural characterization of the rice residues. Four different scenarios were proposed to compare the energy-driven biorefineries based on fast pyrolysis and gasification considering technical, economic, and environmental metrics. These biorefineries were simulated using the Aspen Plus V.14.0 software. The novelty of this research is focused on the identification of the biorefinery with the best techno-economic, energetic, and environmental performance in the Colombian context. Economic and environmental analyses were done by using economic metrics and emissions. From an economic perspective, the stand-alone gasification process did not have a positive economic margin. In contrast, the fast pyrolysis process has the best economic performance since this process has a positive profit margin. Indeed, scenario 1 (fast pyrolysis of both rice residues) presented an economic margin of 13.75% and emissions of 2170.92 kgCO2eq/kg for 10 years. However, this scenario was not energetically the best, holding second place due to the feedstock requirements, compared to gasification. The biorefinery scenario 1 has the best performance.

Design and Development of Learning Management System Huemul for Teaching Fast Healthcare Interoperability Resource: Algorithm Development and Validation Study.

BACKGROUND: Interoperability between health information systems is a fundamental requirement to guarantee the continuity of health care for the population. The Fast Healthcare Interoperability Resource (FHIR) is the standard that enables the design and development of interoperable systems with broad adoption worldwide. However, FHIR training curriculums need an easily administered web-based self-learning platform with modules to create scenarios and questions that the learner answers. This paper proposes a system for teaching FHIR that automatically evaluates the answers, providing the learner with continuous feedback and progress. OBJECTIVE: We are designing and developing a learning management system for creating, applying, deploying, and automatically assessing FHIR web-based courses. METHODS: The system requirements for teaching FHIR were collected through interviews with experts involved in academic and professional FHIR activities (universities and health institutions). The interviews were semistructured, recording and documenting each meeting. In addition, we used an ad hoc instrument to register and analyze all the needs to elicit the requirements. Finally, the information obtained was triangulated with the available evidence. This analysis was carried out with Atlas-ti software. For design purposes, the requirements were divided into functional and nonfunctional. The functional requirements were (1) a test and question manager, (2) an application programming interface (API) to orchestrate components, (3) a test evaluator that automatically evaluates the responses, and (4) a client application for students. Security and usability are essential nonfunctional requirements to design functional and secure interfaces. The software development methodology was based on the traditional spiral model. The end users of the proposed system are (1) the system administrator for all technical aspects of the server, (2) the teacher designing the courses, and (3) the students interested in learning FHIR. RESULTS: The main result described in this work is Huemul, a learning management system for training on FHIR, which includes the following components: (1) Huemul Admin: a web application to create users, tests, and questions and define scores; (2) Huemul API: module for communication between different software components (FHIR server, client, and engine); (3) Huemul Engine: component for answers evaluation to identify differences and validate the content; and (4) Huemul Client: the web application for users to show the test and questions. Huemul was successfully implemented with 416 students associated with the 10 active courses on the platform. In addition, the teachers have created 60 tests and 695 questions. Overall, the 416 students who completed their courses rated Huemul highly. CONCLUSIONS: Huemul is the first platform that allows the creation of courses, tests, and questions that enable the automatic evaluation and feedback of FHIR operations. Huemul has been implemented in multiple FHIR teaching scenarios for health care professionals. Professionals trained on FHIR with Huemul are leading successful national and international initiatives.

From the closet to the shore: Fashion waste pollution on Colombian Central Caribbean beaches.

This paper examines the environmental effects of fashion waste on the central Caribbean coast of Colombia, highlighting significant ecological issues in coastal areas. A survey of 27 beaches found 503 items of fashion waste, mainly shoes and sandals, with an average density of 0.93 items/m2. The distribution of this type of waste varies notably across different beach types, with exposed, rural, and remote beaches showing higher accumulation, indicating a relationship between beach characteristics and waste concentration. Primary sources of this waste include riverine transport, coastal tourism, and poorly managed sewage systems, impacting urban, rural, and village beaches differently. The study reveals that longshore currents and oceanic movements, significantly influence the transport and fate of this waste, with exposed beaches accumulating more fashion waste than sheltered ones. The presence of driftwood also plays a vital role in trapping fashion waste along coastlines. The findings highlight the need for effective management strategies to mitigate the impact of fashion waste, providing crucial insights for local and national coastal managers and implications for managing fashion waste in the Southern Caribbean and similar regions worldwide.

On-Demand ICS + FABA Combinations in 6-11-Year-Old Children.

In recent years, some new concepts have been added to asthma treatment such as "anti-inflammatory reliever" (ß2-agonist use associated to an inhaled corticosteroid (ICS) as a reliever treatment) that combines the benefits of both therapies and provides short- and long-term benefits for treatment in asthma patients. Robust evidence has been presented in patients over 12 years, and the main changes in the international guidelines for asthma treatment were originally made in this age group. However, a few suggestions have been added to treatments in younger patients, in part because of the scarce evidence that exists in this group. We aim to analyze the information regarding the utilization of ICS + fast-acting beta-agonist (FABA) combination in children between 6 and 11 years. Although up until today only three published trials exist (two studies use beclomethasone + albuterol and one study uses budesonide + formoterol), they provide significant information on the benefits of ICS + FABA use in this population.

Fast Parabolic Fitting: An R-Peak Detection Algorithm for Wearable ECG Devices.

Heart diseases rank among the most fatal health concerns globally, with the majority being preventable through early diagnosis and effective treatment. Electrocardiogram (ECG) analysis is critical in detecting heart diseases, as it captures the heart's electrical activities. For continuous monitoring, wearable electrocardiographic devices must ensure user comfort over extended periods, typically 24 to 48 h. These devices demand specialized algorithms with low computational complexity to accommodate memory and power consumption constraints. One of the most crucial aspects of ECG signals is accurately detecting heartbeat intervals, specifically the R peaks. In this study, we introduce a novel algorithm designed for wearable devices, offering two primary attributes: robustness against noise and low computational complexity. Our algorithm entails fitting a least-squares parabola to the ECG signal and adaptively shaping it as it sweeps through the signal. Notably, our proposed algorithm eliminates the need for band-pass filters, which can inadvertently smooth the R peaks, making them more challenging to identify. We compared the algorithm's performance using two extensive databases: the meta-database QT database and the BIH-MIT database. Importantly, our method does not necessitate the precise localization of the ECG signal's isoelectric line, contributing to its low computational complexity. In the analysis of the QT database, our algorithm demonstrated a substantial advantage over the classical Pan-Tompkins algorithm and maintained competitiveness with state-of-the-art approaches. In the case of the BIH-MIT database, the performance results were more conservative; they continued to underscore the real-world utility of our algorithm in clinical contexts.

A controlled evaluation of filter paper use during staining of sputum smears for tuberculosis microscopy.

Background: Some sputum smear microscopy protocols recommend placing filter paper over sputum smears during staining for Mycobacterium tuberculosis (TB) . We found no published evidence assessing whether this is beneficial. We aimed to evaluate the effect of filter paper on sputum smear microscopy results. Methods: Sputum samples were collected from 30 patients with confirmed pulmonary TB and 4 healthy control participants. From each sputum sample, six smears (204 smears in total) were prepared for staining with Ziehl-Neelsen (ZN), auramine or viability staining with fluorescein diacetate (FDA). Half of the slides subjected to each staining protocol were randomly selected to have Whatman grade 3 filter paper placed over the dried smears prior to stain application and removed prior to stain washing. The counts of acid-fast bacilli (AFB) and precipitates per 100 high-power microscopy fields of view, and the proportion of smear that appeared to have been washed away were recorded. Statistical analysis used a linear regression model adjusted by staining technique with a random effects term to correct for between-sample variability.   Results: The inclusion of filter paper in the staining protocol significantly decreased microscopy positivity independent of staining with ZN, auramine or FDA (p=0.01). Consistent with this finding, there were lower smear grades in slides stained using filter paper versus without (p=0.04), and filter paper use reduced AFB counts by 0.28 logarithms (95% confidence intervals, CI=0.018, 0.54, p=0.04) independent of staining technique. In all analyses, auramine was consistently more sensitive with higher AFB counts versus ZN (p=0.001), whereas FDA had lower sensitivity and lower AFB counts (p<0.0001). Filter paper use was not associated with the presence of any precipitate (p=0.5) or the probability of any smear washing away (p=0.6) during the staining process. Conclusions: Filter paper reduced the sensitivity of AFB microscopy and had no detectable beneficial effects so is not recommended.

Marsupial position on life-history continua and the potential contribution of life-history traits to population growth.

Previous studies have suggested that mammal life history varies along the fast-slow continuum and that, in eutherians, this continuum is linked to variation in the potential contribution of survival and reproduction to population growth rate (λ). Fast eutherians mature early, have large litters and short lifespans, and exhibit high potential contribution of age at first reproduction and fertility to λ, while slow eutherians show high potential contribution of survival to λ. However, marsupials have typically been overlooked in comparative tests of mammalian life-history evolution. Here, we tested whether the eutherian life-history pattern extends to marsupials, and show that marsupial life-history trade-offs are organized along two major axes: (i) the reproductive output and dispersion axis, and (ii) the fast-slow continuum, with an additional association between adult survival and body mass. Life-history traits that potentially drive changes in λ are similar in eutherians and marsupials with slow life histories, but differ in fast marsupials; age at first reproduction is the most important trait contributing to λ and fertility contributes little. Marsupials have slower life histories than eutherians, and differences between these clades may derive from their contrasting reproductive modes; marsupials have slower development, growth and metabolism than eutherians of equivalent size.

Development and Implementation of an Enhanced Recovery Protocol for Bariatric Patients in a Third World Environment.

Introduction: An applicable and reproducible enhanced recovery protocol was developed and implemented to improve our outcomes in a third-world environment. Methods: We compared the results obtained prospectively. The group treated before the application of the enhanced recovery protocol was called usual care (UC) and included all bariatric surgeries operated on between 2014 and 2017. The new protocol was applied between 2017 and 2019 including all operated patients, and this group was called Fast Track (FT). The variables analyzed were the length of stay, readmissions, and complications recorded during the first 30 days. We also analyzed the milligrams of morphine used by each patient, and a cost analysis was performed. Results: During the study period, 816 patients were studied. Of these, 385 (47.2%) belonged to the UC group and 431 (52.8%) to the FT group. The mean hospital stay was 58.5 hours (UC) versus 40.3 hours (FT) (P = .0001). When comparing the global morbidity of both groups, we did not find significant differences (P = .47). There was also no statistically significant difference when comparing major complications (P = .79). No mortality was recorded. Morphine indication reported a statistically significant difference that favored FT. Costs were significantly higher in UC than in FT (P < .0001). Conclusions: We believe that the implementation of an enhanced recovery protocol in bariatric surgery is a reliable measure and can be implemented even in an underdevelopment environment enlarging the benefit for patients.

Rational PCR Reactor Design in Microfluidics.

Limit of detection (LOD), speed, and cost for some of the most important diagnostic tools, i.e., lateral flow assays (LFA), enzyme-linked immunosorbent assays (ELISA), and polymerase chain reaction (PCR), all benefited from both the financial and regulatory support brought about by the pandemic. From those three, PCR has gained the most in overall performance. However, implementing PCR in point of care (POC) settings remains challenging because of its stringent requirements for a low LOD, multiplexing, accuracy, selectivity, robustness, and cost. Moreover, from a clinical point of view, it has become very desirable to attain an overall sample-to-answer time (t) of 10 min or less. Based on those POC requirements, we introduce three parameters to guide the design towards the next generation of PCR reactors: the overall sample-to-answer time (t); lambda (λ), a measure that sets the minimum number of copies required per reactor volume; and gamma (γ), the system's thermal efficiency. These three parameters control the necessary sample volume, the number of reactors that are feasible (for multiplexing), the type of fluidics, the PCR reactor shape, the thermal conductivity, the diffusivity of the materials used, and the type of heating and cooling systems employed. Then, as an illustration, we carry out a numerical simulation of temperature changes in a PCR device, discuss the leading commercial and RT-qPCR contenders under development, and suggest approaches to achieve the PCR reactor for RT-qPCR of the future.

Focused Assessment with Sonography for Trauma (FAST).

The main cause of death in traumas is hypovolemic shock. Physical examination is limited to detect hemopericardium, hemoperitoneum, and hemopneumothorax. Computed tomography (CT) is the gold standard for traumatic injury evaluation. However, CT is not always available, is more expensive, and there are transportation issues, especially in hemodynamically unstable patients. In this scenario, a rapid, reproducible, portable, and noninvasive method such as ultrasound emerged, directed for detecting hemopericardium, hemoperitoneum, and hemopneumothorax, in a "point of care" modality, known as the focused assessment with sonography for trauma (FAST) protocol. With decades of experience, spread worldwide, and recommended by the most prestigious trauma care guidelines, FAST is a bedside ultrasound to be performed when accessing circulation issues of trauma patients. It is indicated to hemodynamically unstable patients with blunt abdominal trauma, with penetrating trauma of the thoracoabdominal transition (where there is doubt of penetrating the abdominal cavity) and for any patient with the cause of the instability unknown. There are four regions to be examined in the traditional FAST protocol: pericardium (to detect cardiac tamponade), right upper abdominal quadrant, left upper abdominal quadrant, and pelvis (to detect hemoperitoneum). The called extended FAST (e-FAST) protocol also searches the pleural spaces for hemothorax and pneumothorax. It is important to know the false positives and false negatives of the protocol, as well as its limitations. FAST/e-FAST protocol is designed to provide a simple "yes or no" answer regarding the presence of bleeding. It is not intended to quantify the bleeding nor evaluate organ lesions due to its limited accuracy for these purposes. Moreover, the amount of bleeding and/or the identification of organ lesions will not change patient's management: Hemodynamically unstable patients with positive FAST must go to the operating room without delay. CT should be considered for hemodynamically stable patients.

A new approach with dried plasma spots: a rapid determination of antidepressants and antipsychotics by fast GC-MS.

Background: The increasing prevalence of poisoning cases related to antidepressants and antipsychotics has raised concerns. Methods: To address this issue, a new adaptation of the dried plasma spot technique was developed using a 24-well plate and fast gas chromatography-mass spectrometry. The method involves the optimization of extraction variables and sample preparation, and was successfully validated. Results: The limits of quantitation ranged from 20 to 60 ng/ml, and accuracy ranged from 87.8% to 112.2%. The technique was applied to 102 human plasma samples from suspected poisoning cases, with positivity of 90.2%. Conclusion: This method provides a cheap, easy to implement and fast approach, making it ideal for toxicological emergency laboratories and promoting valuable support for healthcare professionals managing poisoning cases involving antidepressants and antipsychotics.

CBIR-SAR System Using Stochastic Distance.

This article proposes a system for Content-Based Image Retrieval (CBIR) using stochastic distance for Synthetic-Aperture Radar (SAR) images. The methodology consists of three essential steps for image retrieval. First, it estimates the roughness (α^) and scale (γ^) parameters of the GI0 distribution that models SAR data in intensity. The parameters of the model were estimated using the Maximum Likelihood Estimation and the fast approach of the Log-Cumulants method. Second, using the triangular distance, CBIR-SAR evaluates the similarity between a query image and images in the database. The stochastic distance can identify the most similar regions according to the image features, which are the estimated parameters of the data model. Third, the performance of our proposal was evaluated by applying the Mean Average Precision (MAP) measure and considering clippings from three radar sensors, i.e., UAVSAR, OrbiSaR-2, and ALOS PALSAR. The CBIR-SAR results for synthetic images achieved the highest MAP value, retrieving extremely heterogeneous regions. Regarding the real SAR images, CBIR-SAR achieved MAP values above 0.833 for all polarization channels for image samples of forest (UAVSAR) and urban areas (ORBISAR). Our results confirmed that the proposed method is sensitive to the degree of texture, and hence, it relies on good estimates. They are inputs to the stochastic distance for effective image retrieval.

Lower Extremity Nodules After Spelunking in Mexico.

Mycobacterium marinum is a non-tuberculous mycobacterium that presents as a nodular granulomatous disease. The bacillus can infect humans when broken skin is exposed to a contaminated aquatic environment. M. marinum infections are usually isolated to the skin and soft tissues and can spread in a lymphatic distribution. A 26-year-old male cut his right ankle while spelunking in Tulum, Mexico. He presented to his primary care physician three months after he sustained the laceration with a nonhealing wound on the right lateral posterior ankle. Examination of the lesion demonstrated erythematous, violaceous, and hyperpigmented indurated plaques with satellite lesions noted at the right medial, posterior, and lateral ankle. The lesion characteristics raised initial suspicion for an invasive fungal infection. Biopsy of the lesion demonstrated epidermal ulceration covered by neutrophilic serum, marked underlying dermal acute inflammation, and granulation tissue. A mild perivascular, predominantly lymphocytic infiltrate was present in the deep dermis with no evidence of granuloma. Acid-fast bacilli culture plated onto chocolate agar confirmed the species M. marinum.

Contenido de sodio en preparaciones de consumo frecuente fuera del hogar en tres zonas de Lima Metropolitana, Perú / Sodium content in commonly consumed away-from-home food in three areas of Metropolitan Lima, Peru

El objetivo fue caracterizar el contenido de sodio en preparaciones de consumo frecuente fuera del hogar en tres zonas de Lima Metropolitana. Se realizó un estudio transversal para el cual se identificaron veinte preparaciones de consumo frecuente según el lugar de expendio. La determinación del contenido de sodio se obtuvo mediante espectroscopía de absorción atómica en preparaciones recolectadas en el 2019. La mediana del contenido de sodio en productos de venta ambulatoria fue 492,36 mg/100 g (RIQ: 83,93 - 918,78); 471,37 mg/100 g, en preparaciones tradicionales y típicas (RIQ: 76,04 - 765,39); y 471,06 mg/100 g (RIQ: 115,31 - 695,18), en comidas rápidas. El 65% de las preparaciones fueron consideradas altas en sodio según los parámetros peruanos, mientras que el 30% de las preparaciones también presentaron un alto contenido, de acuerdo con los parámetros del Reino Unido. La mayoría de las preparaciones expendidas y consumidas fuera del hogar presentan un elevado contenido de sodio. Es esencial involucrar a todos los actores que participan en la preparación de alimentos para el consumo fuera del hogar, con el fin de sensibilizarlos e incluirlos en la promoción de políticas enfocadas en la reducción del consumo de sodio.

The aim of this study was to characterize the sodium content in commonly consumed away-from-home food in three areas of Metropolitan Lima. We conducted a cross-sectional study, in which twenty frequently consumed foods were identified according to the place of sale. Sodium content was determined through atomic absorption spectroscopy in preparations collected in 2019. The median sodium content in street food products was 492.36 mg/100g (IQR: 83.93 - 918.78), 471.37 mg/100 g in artisanal food (IQR: 76.04 - 765.39) and 471.06 mg/100 g in fast food (IQR: 115.31 - 695.18). Sixty-five percent of the foods were classified as having high sodium content according to Peruvian regulations, while 30% of the preparations had high sodium content, according to UK parameters. Most food sold and consumed away from home have high sodium content. It is essential to engage all stakeholders involved in food preparation for away-from-home consumption in order to raise awareness and involve them in the promotion of policies aimed at reducing sodium intake.

Sample pooling and incurred samples improve analytical throughput and quality control of lipophilic phycotoxins screening in bivalve mollusks.

Lipophilic marine biotoxins (LMBs) are one of the main risks associated with the consumption of mussels and oysters. Sanitary and analytical control programs are developed to detect the occurrence of these toxins in seafood before they reach toxic levels. To ensure quick results, methods must be easy and fast to perform. In this work, we demonstrated that incurred samples were a viable alternative to validation and internal quality control studies for the analysis of LMBs in bivalve mollusks. These samples were used to optimize, validate, and monitor a simple and fast ultrasound-assisted extraction (UAE) procedure. An internal quality control material containing okadaic acid (227 ± 46 µg kg-1) was produced and characterized. This material had its homogeneity and stability verified and was included as a quality control in all batches of analytical routine. Besides, a sample pooling protocol for extracts analysis was developed, based on tests for COVID-19. Up to 10 samples could be analyzed simultaneously, reducing the instrumental time of analysis by up to 80%. The UAE and sample pooling approaches were then applied to more than 450 samples, of which at least 100 were positive for the okadaic acid group of toxins.