Cell Culture Differentiation and Proliferation Conditions Influence the In Vitro Regeneration of the Human Airway Epithelium.

The human airway mucociliary epithelium can be recapitulated in vitro using primary cells cultured in an Air-Liquid Interface (ALI), a reliable surrogate to perform pathophysiological studies. As tremendous variations exist between media used for ALI-cultured human airway epithelial cells, our study aimed to evaluate the impact of several media (BEGMTM, PneumaCultTM, "Half&Half" and "Clancy") on cell type distribution using single-cell RNA sequencing and imaging. Our work revealed the impact of these media on cell composition, gene expression profile, cell signaling and epithelial morphology. We found higher proportions of multiciliated cells in PneumaCultTM-ALI and Half&Half, stronger EGF signaling from basal cells in BEGMTM-ALI, differential expression of the SARS-CoV-2 entry factor ACE2, and distinct secretome transcripts depending on media used. We also established that proliferation in PneumaCultTM-Ex Plus favored secretory cell fate, showing the key influence of proliferation media on late differentiation epithelial characteristics. Altogether, our data offer a comprehensive repertoire for evaluating the effects of culture conditions on airway epithelial differentiation and will help to choose the most relevant medium according to the processes to be investigated such as cilia, mucus biology or viral infection. We detail useful parameters that should be explored to document airway epithelial cell fate and morphology. This article is open access and distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Adaptation and Psychometric Properties of an Attitude toward Artificial Intelligence Scale (AIAS-4) among Peruvian Nurses.

BACKGROUND: The integration of Artificial Intelligence (AI) into various aspects of daily life has sparked growing interest in understanding public attitudes toward this technology. Despite advancements in tools to assess these perceptions, there remains a need for culturally adapted instruments, particularly in specific contexts like that of Peruvian nurses. OBJECTIVE: To evaluate the psychometric properties of the AIAS-4 in a sample of Peruvian nurses. METHODS: An instrumental design was employed, recruiting 200 Peruvian nurses. The Attitude toward Artificial Intelligence in Spanish (AIAS-S), a cultural and linguistic adaptation of the AIAS-4, involved data analysis using descriptive statistics, confirmatory factor analysis (CFA), and invariance tests. RESULTS: The Confirmatory Factor Analysis (CFA) confirmed a unidimensional factor structure with an excellent model fit (χ2 = 0.410, df = 1, p = 0.522, CFI = 1.00, TLI = 1.00, RMSEA = 0.00, SRMR = 0.00). The scale demonstrated high internal consistency (α = 0.94, ω = 0.91). Tests of invariance from configural to strict confirmed that the scale is stable across different demographic subgroups. CONCLUSIONS: The AIAS-S proved to be a psychometrically solid tool for assessing attitudes toward AI in the context of Peruvian nurses, providing evidence of validity, reliability, and gender invariance. This study highlights the importance of having culturally adapted instruments to explore attitudes toward emerging technologies in specific groups.

Developing and validating a prediction model of live birth following single vitrified-warmed blastocyst transfer.

RESEARCH QUESTION: Can the developed clinical prediction model offer an accurate estimate of the likelihood of live birth, involving blastocyst morphology and vitrification day after single vitrified-warmed blastocyst transfer (SVBT), and therefore assist clinicians and patients? STUDY DESIGN: Retrospective cohort study conducted at a Spanish university-based reproductive medicine unit (2017-2021) including consecutive vitrified-warmed blastocysts from IVF cycles. A multivariable logistic regression incorporated key live birth predictors: vitrification day, embryo score, embryo ploidy status and clinically relevant variables, i.e. maternal age. RESULTS: The training set involved 1653 SVBT cycles carried out between 2017 and 2020; 592 SVBT cycles from 2021 constituted the external validation dataset. The model revealed that female age and embryo characteristics, including overall quality and blastulation day, is linked to live birth rate in SVBT cycles. Stratification by vitrification day and quality (from day-5A to day-6 C blastocysts) applied to genetically tested and untested embryos. The model's area under the curve was 0.66 (95% CI 0.64 to 0.69) during development and 0.65 (95% CI 0.61 to 0.70) in validation, denoting moderate discrimination. Calibration plots showed strong agreement between predicted and observed probabilities. CONCLUSION: By incorporating essential predictors such as vitrification day, embryo morphology grade, age and preimplantation genetic testing for aneuploidy usage, this predictive model offers valuable guidance to clinicians and patients, enabling accurate forecasts of live birth rates for any given vitrified blastocyst within SVBT cycles. Additionally, it serves as a potentially indispensable laboratory tool, aiding in selecting the most promising blastocysts for optimal outcomes.

Safety and efficacy of continuous terlipressin infusion in HRS-AKI in a transplant population.

Hepatorenal syndrome-acute kidney injury (HRS-AKI) is associated with significant morbidity and mortality. While liver transplantation is the definitive treatment, continuous terlipressin infusion for HRS-AKI may provide benefit and, as such, was assessed in a population composed of candidates for liver transplant (LT). Fifty hospitalized LT-eligible patients with HRS-AKI received a single bolus followed by continuous terlipressin infusion. Acute-on-chronic liver failure grade 3, serum creatinine (SCr)>5.0 mg/dL, or Model for End-Stage Liver Disease (MELD) ≥35 were exclusions. Fifty hospitalized patients who received midodrine and octreotide or norepinephrine for HRS-AKI served as a historical comparator cohort. Complete response (CR) was defined as a ≥30% decrease in SCr with end-of-treatment (EOT) SCr≤1.5, partial response as a ≥30% decrease in SCr with EOT SCr>1.5, and nonresponse as a <30% decrease in SCr. CR rate was significantly higher in the terlipressin cohort compared to the historical cohort (64% vs. 16%, p <0.001). Survival, while numerically higher in those who received terlipressin, was statistically similar (D30: 94% vs. 82%, p =0.12; D90: 78% vs. 68%, p =0.37). Renal replacement therapy (RRT) was more common among terlipressin NR than CR and PR (70% vs. 3% vs. 13%, p < 0.001). EOT MELD and SCr were significantly lower within terlipressin cohort (MELD: 19 vs. 25, SCr: 1.4 vs. 2.1 mg/dL, p <0.001). Sixteen of 40 terlipressin-treated patients received LT-alone (terlipressin CR in 10/16). One patient on terlipressin had a hypoxic respiratory failure that responded to diuretics; one possibly had drug-related rash. With continuous terlipressin infusion, a CR rate of 64% was observed with a favorable safety profile. Terlipressin use was associated with lower EOT MELD and SCr than the historical midodrine and octreotide/norepinephrine cohort; LT-alone was accomplished in a high proportion of complete terlipressin responders.

Usability evaluation of the educational website "understanding my diabetes" for Mexican patients with type 2 diabetes.

Background: Diabetes education is an integral part of the treatment for the metabolic control of patients with diabetes. The use of the Internet as a tool for diabetes education, as well as its acceptance, is still under study. Aim: To assess the usability of the educational website "I understand my diabetes" designed for patients with type 2 diabetes attending primary care clinics. Material and method: A cross-sectional study was done in 110 patients with type 2 diabetes from two family medicine clinics, each of whom was assigned a user account on the educational website "Entiendo mi diabetes." The web site assigned a user name and password to each patient. They were able to access the educational website at home. After a 15-day review period, participants were asked to evaluate usability using the Computer System Usability Questionnaire. Additionally, we developed an eight-item questionnaire usability focusing on diabetes care. Sociodemographic data, blood pressure, and anthropometric measurements were recorded. Glucose levels and lipid profiles were also measured. Results: The patients with diabetes had a mean age of 52.7 years and a median of 5 years since they were diagnosed with diabetes. The website received a good usability rating from 89.1% of participants, with favorable assessments in all three dimensions: 87.3% for information, 85.5% for quality, and 88.2% for interface. Regarding usability specifically for diabetes care, 98.2% rated it as having good usability. Conclusion: The website for education about the disease in patients "I understand my diabetes" had an adequate usability evaluation by patients, so they also considered it very useful for diabetes care. The diabetes care instrument had adequate usability and reliability.

Association between cardiovascular risk and maternal perception of BMI in Peruvian schoolchildren.

Introduction: In the modern era, the maternal perception of children's nutritional status has emerged as a critical area of study, given its potential influence on nutritional interventions and long-term child health. The relationship between this perception and children's Body Mass Index (BMI) by age is particularly intriguing, as it may reveal discrepancies between perception and reality. Objective: The aim of this study was to evaluate Peruvian mothers' perception of their children's Body Mass Index (BMI) in relation to age and to determine how this perception associates with the children's cardiovascular risk. The study also analyzed sociodemographic factors that might influence this perception. Methods: The study included 130 mothers of schoolchildren aged 5 to 11 from a school in Lima. Mothers' perceptions of their children's weight were assessed using pictograms, and sociodemographic characteristics were collected through a questionnaire. Weight and height measurements were taken to calculate BMI, and waist circumference was measured to classify cardiovascular risk. Results: A total of 57.4% of the schoolchildren presented with excess malnutrition, and 51.5% of the mothers incorrectly classified the actual BMI/Age of their children (kappa 0.11; p ≤ 0.05). Additionally, it was found that the schoolchild's age is associated with the mother's failure to accurately perceive her child's weight (OR 1.59). Lastly, there was a significant association between maternal perception and cardiovascular risk (p ≤ 0.05). Conclusion: There is a significant discrepancy between maternal perception and the actual nutritional status of children, which can increase cardiovascular risk. It is necessary to implement intervention and education strategies targeted at parents to enhance the recognition and management of childhood overweight and obesity.

Psychometric Properties of a Short Academic Motivation Scale (SAMS) in Medical Students.

Background: Medical education represents a complex field of study, influenced by various psychological, demographic, and contextual factors. Academic motivation, essential for educational success, has been linked to critical decisions in medical careers and can be modulated by contextual elements such as socioeconomic and geographical environments. The theory of self-determination has provided a solid framework for understanding the multidimensional nature of motivation. Objective: To evaluate the psychometric properties of a Spanish version of the Short Scale of Academic Motivation among Peruvian medical students. Methods: Using an instrumental design, the factorial structure, reliability, and gender invariance of the SAMS-S were assessed. A confirmatory factor analysis (CFA) was conducted to validate the scale's structure based on seven dimensions. Additionally, reliability was assessed through Cronbach's alpha coefficient and omega coefficient, and gender invariance was determined through multi-group confirmatory factor analysis. Results: The Peruvian version of the SAMS-S showed a good fit in the CFA with satisfactory goodness-of-fit indices. However, challenges in discriminant validity among certain dimensions were detected, suggesting the presence of a second-order factor. The proposed second-order model yielded an adequate fit (χ2 = 198.26, df = 70, CFI = 0.92, TLI = 0.90, RMSEA = 0.08 [90% CI: 0.07-0.1], SRMR = 0.07), validating the factorial structure of the SAMS-S. The scale's reliability and its subscales were within acceptable ranges. Furthermore, the gender invariance of the SAMS-S was confirmed at all levels, from configural to strict. Conclusions: The second-order model of the SAMS-S presents as a valid and reliable tool for measuring academic motivation among medical students in Peru. Its robustness and adaptability make it relevant for future research in similar educational contexts and can serve as a basis for interventions aimed at improving academic motivation in this specific group.

Epidemiology, clinical features, and classification of 3,404 patients with uveitis: Colombian Uveitis Multicenter Study (COL-UVEA).

PURPOSE: To describe the epidemiology, clinical features, and classification of uveitis in a large cohort of Colombian patients. METHODS: Data were collected from seven ophthalmological referral centers in the four main cities in Colombia. The study included patients with a confirmed diagnosis of uveitis from January 2010 to December 2022. Information on demographics, ophthalmic examination findings, uveitis classification, and etiology was recorded. RESULTS: The study reviewed 3,404 clinical records of patients with uveitis. The mean age at diagnosis was 41.1 (SD 19.0) years, and 54.2% of the patients were female. Overall, 1,341(39.4%) were infectious, 626 (18.4%) non-infectious, and four masquerade syndromes (0.1%). The most common types of uveitis were unilateral (66.7%), acute (48.3%), and non-granulomatous (83%). Anterior uveitis was the most common anatomical localization (49.5%), followed by posterior uveitis (22.9%), panuveitis (22.3%), and intermediate uveitis (5.2%). A diagnosis was established in 3,252 (95.5%) cases; idiopathic was the most common cause (27.7%), followed by toxoplasmosis (25.3%) and virus-associated uveitis (6.4%). The age group between 30 and 50 exhibited the highest frequency of uveitis. CONCLUSION: This multicenter study comprehensively describes uveitis characteristics in Colombian patients, providing valuable insights into its demographic and clinical features. The study findings emphasize the need to continue updating the changing patterns of uveitis to improve diagnosis and treatment strategies for diseases associated with intraocular inflammation.

Analytical and functional profiles of paralytic shellfish toxins extracted from Semele proficua and Senilia senilis from Angola.

Paralytic shellfish poisoning is a foodborne illness that typically derive from the consumption of shellfish contaminated with saxitoxin-group of toxins produced by dinoflagellates of the genus Gymnodinium, Alexandrium and Pyrodinium. N-sulfocarbamoyl, carbamate and dicarbamoyl are the most abundant. In 2007 and 2008 some episodes of PSP occurred in Angola where there is not monitoring program for shellfish contamination with marine biotoxins. Therefore, ten samples extracted from Semele proficua from Luanda Bay and Senilia senilis from Mussulo Bay, were analyzed by HPLC finding saxitoxin, decarbamoylsaxitoxin and other three compounds that have an unusual profile different to the known hydrophilic PSP toxins were found in different amounts and combinations. These new compounds were not autofluorescent, and they presented much stronger response after peroxide oxidation than after periodate oxidation. The compounds appear as peaks eluted at 2.5 and 5.6 min after periodate oxidation and 8.2 min after peroxide oxidation. Electrophysiological studies revealed that none of the three unknown compounds had effect at cellular level by decreasing the maximum peak inward sodium currents by blocking voltage-gated sodium channels. Thus, not contributing to PSP intoxication. The presence in all samples of saxitoxin-group compounds poses a risk to human health and remarks the need to further explore the presence of new compounds that contaminate seafood, investigating their activity and developing monitoring programs.

Blastocysts from partial compaction morulae are not defined by their early mistakes.

RESEARCH QUESTION: Is partial compaction during morula formation associated with an embryo's developmental ability and implantation potential? DESIGN: Retrospective analysis of data from 196 preimplantation genetic testing for aneuploidy (PGT-A) cycles. Embryos starting compaction were grouped according to the inclusion or not of all the blastomeres in the forming morula (full compaction or partial compaction). The possible effect of maternal age and ovarian response on compaction was analysed. Morphokinetic characteristics, blastocyst formation rate, morphology and cytogenetic constitution of the obtained blastocysts were compared. Comparisons of reproductive outcomes after the transfer of euploid blastocysts from both groups were established. Finally, in a subset of embryos, the chromosomal constitution concordance of the abandoned cells and the corresponding blastocyst through trophectoderm biopsies was assessed. RESULTS: A total of 430 embryos failed to include at least one cell during compaction (partial compaction group [49.3%]), whereas the 442 remaining embryos formed a fully compacted morula (full compaction group [50.7%]). Neither female age nor the number of oocytes collected affected the prevalence of partial compaction morulae. Morphokinetic parameters were altered in embryos from partial compaction morulae compared with full compaction. Although an impairment in blastocyst formation rate was observed in partial compaction morulae (57.2% versus 70.8%, P < 0.001), both chromosomal constitution (euploidy rate: partial compaction [38.4%] versus full compaction [34.2%]) and reproductive outcomes (live birth rate: partial compaction [51.9%] versus full compaction [46.2%]) of the obtained blastocysts were equivalent between groups. A high ploidy correlation of excluded cells-trophectoderm duos was observed. CONCLUSIONS: Partial compaction morulae show a reduced developmental ability compared with full compaction morulae. Resulting blastocysts from both groups, however, have similar euploidy rates and reproductive outcomes. Cell exclusion might be a consequence of a compromised embryo development regardless of the chromosomal constitution of the excluded cells.

Depression, professional self-efficacy, and job performance as predictors of life satisfaction: the mediating role of work engagement in nurses.

Background: The life satisfaction and job performance of nursing professionals are affected by a multitude of factors, including work engagement, self-efficacy, and depression. The Job Demands-Resources (JD-R) model provides a theoretical framework to explore these relationships. Objective: Our study aimed to analyze the primary goal of this research, which is to examine the mediating role of work engagement in the relationship between depression, professional self-efficacy, job performance, and their impact on life satisfaction in nurses, using the JD-R theory as a guide. Methods: This cross-sectional study involved 579 participants aged between 21 to 57 years (M = 39, SD = 9.95). Mediation analysis was used to examine the influence of depression, self-efficacy, and job performance on work engagement, and in turn, its effect on life satisfaction. Results: Findings indicated that work engagement plays a crucial mediating role between depression, self-efficacy, job performance, and life satisfaction. Interventions to increase work engagement could assist nurses in better managing depression and improving their performance and life satisfaction. Conclusions: Our study highlights the need for workplace policies and strategies that foster work engagement and self-efficacy among nurses while effectively managing job demands to prevent depression. Moreover, these findings underscore the importance of the JD-R theory to understand and improve nurses' job satisfaction and performance, and suggest areas for future research, including exploring other potential factors and applying these findings across different contexts and cultures.

Surgical Outcomes in Uveitic Glaucoma: Long-Term Evaluation of Trabeculectomy, Non-Penetrating Deep Sclerectomy, Ex-PRESS Shunt and Ahmed Glaucoma Valve. A 3-Year Follow-Up Study.

PURPOSE: Uveitis can lead to secondary glaucoma, a condition with challenging management that can carry irreversible visual loss. Filtering surgery has demonstrated a higher failure rate, increased incidence of postoperative complications and reinterventions in uveitic patients. There is no consensus on the optimal surgical approach for uveitic glaucoma (UG) due to limited data comparing various intraocular pressure (IOP)-lowering surgeries. This retrospective cohort aims to assess the clinical outcomes of trabeculectomy (TBT), non-penetrating deep sclerectomy (NPDS), Ex-PRESS shunt and Ahmed glaucoma valve, providing additional insights into the long-term IOP control and safety of filtering surgeries in UG. METHODS: The filtering surgery was performed on 32 eyes of 27 UG patients. Complete success was defined as IOP ≤ 18 mmHg or a 30% reduction. Qualified success allowed topical hypotensive treatment. RESULTS: Complete success was 40.63% (13/32) at 12 months and 36.67% (11/30) at 36 months. Qualified success was 84.38% (27/32) at 12 months and 63.33% (19/30) at 36 months. In the survival analysis, both NPDS and Ex-PRESS demonstrated decreased failure rates compared to TBT (NPDS vs TBT: HR = 0.20, p = 0.049; Ex-PRESS vs TBT: HR = 0.28, p = 0.13). One or more reinterventions were required in 34.38% (11/32) of the eyes. NPDS had the lowest incidence of hypotony. Secondary cataract was a common complication in all groups. CONCLUSION: Various filtering surgeries are safe and effective procedures for lowering IOP and reducing the requirement of topical antihypertensives in UG at 36 months. However, one-third of the patients will require another IOP-lowering procedure.

Exosomal Osteoclast-Derived miRNA in Rheumatoid Arthritis: From Their Pathogenesis in Bone Erosion to New Therapeutic Approaches.

Rheumatoid arthritis (RA) is an autoimmune disease that causes inflammation, pain, and ultimately, bone erosion of the joints. The causes of this disease are multifactorial, including genetic factors, such as the presence of the human leukocyte antigen (HLA)-DRB1*04 variant, alterations in the microbiota, or immune factors including increased cytotoxic T lymphocytes (CTLs), neutrophils, or elevated M1 macrophages which, taken together, produce high levels of pro-inflammatory cytokines. In this review, we focused on the function exerted by osteoclasts on osteoblasts and other osteoclasts by means of the release of exosomal microRNAs (miRNAs). Based on a thorough revision, we classified these molecules into three categories according to their function: osteoclast inhibitors (miR-23a, miR-29b, and miR-214), osteoblast inhibitors (miR-22-3p, miR-26a, miR-27a, miR-29a, miR-125b, and miR-146a), and osteoblast enhancers (miR-20a, miR-34a, miR-96, miR-106a, miR-142, miR-199a, miR-324, and miR-486b). Finally, we analyzed potential therapeutic targets of these exosomal miRNAs, such as the use of antagomiRs, blockmiRs, agomiRs and competitive endogenous RNAs (ceRNAs), which are already being tested in murine and ex vivo models of RA. These strategies might have an important role in reestablishing the regulation of osteoclast and osteoblast differentiation making progress in the development of personalized medicine.

Prevalence of corpus callosum pathology in an unselected population. Should assessment of the corpus callosum be included in the routine 20 weeks scan?

OBJECTIVES: To determine the prevalence of abnormalities of the corpus callosum (AbnCC) in a non-selected population, to propose a systematic screening protocol for AbnCC in all populations through direct assessment, and to describe the follow-up and prognosis of all AbnCC cases diagnosed in our clinical setting. METHODS: This was a retrospective review of the prevalence of AbnCC over 11 years. We included a sagittal assessment of the corpus callosum (CC) in the second-trimester scan. AbnCC was classified into complete agenesis of CC (ACC) and dysgenesis of CC (DCC; including small, partial agenesis, thick and with lipoma). RESULTS: Of the 38,586 second-trimester scans performed during our screening, 43 cases of AbnCC were detected (prevalence of 0.8/1000). Of the AbnCC cases, 10 cases were identified as ACC (29.40%) and 24 as DCC (70.59%). Follow-up investigations showed that in the 43 cases with AbnCC, 76.5% had other associated ultrasound abnormalities, 26.5% had genetic abnormalities, 11.8% had other MRI abnormalities, and 25% of the children had neurodevelopmental delays (8.8% of the total), which were severe in only one case. CONCLUSIONS: AbnCC is found in approximately 0.8/1000 of cases in an unselected population. The findings suggest that systematic and direct assessment of the CC as part of screening ultrasound in the second trimester of gestation should be recommended as a routine practice.

Urease-powered nanobots for radionuclide bladder cancer therapy.

Bladder cancer treatment via intravesical drug administration achieves reasonable survival rates but suffers from low therapeutic efficacy. To address the latter, self-propelled nanoparticles or nanobots have been proposed, taking advantage of their enhanced diffusion and mixing capabilities in urine when compared with conventional drugs or passive nanoparticles. However, the translational capabilities of nanobots in treating bladder cancer are underexplored. Here, we tested radiolabelled mesoporous silica-based urease-powered nanobots in an orthotopic mouse model of bladder cancer. In vivo and ex vivo results demonstrated enhanced nanobot accumulation at the tumour site, with an eightfold increase revealed by positron emission tomography in vivo. Label-free optical contrast based on polarization-dependent scattered light-sheet microscopy of cleared bladders confirmed tumour penetration by nanobots ex vivo. Treating tumour-bearing mice with intravesically administered radio-iodinated nanobots for radionuclide therapy resulted in a tumour size reduction of about 90%, positioning nanobots as efficient delivery nanosystems for bladder cancer therapy.

Circadian regulation of MGMT expression and promoter methylation underlies daily rhythms in TMZ sensitivity in glioblastoma.

BACKGROUND: Glioblastoma (GBM) is the most common primary brain tumor in adults. Despite extensive research and clinical trials, median survival post-treatment remains at 15 months. Thus, all opportunities to optimize current treatments and improve patient outcomes should be considered. A recent retrospective clinical study found that taking TMZ in the morning compared to the evening was associated with a 6-month increase in median survival in patients with MGMT-methylated GBM. Here, we hypothesized that TMZ efficacy depends on time-of-day and O6-Methylguanine-DNA Methyltransferase (MGMT) activity in murine and human models of GBM. METHODS AND RESULTS: In vitro recordings using real-time bioluminescence reporters revealed that GBM cells have intrinsic circadian rhythms in the expression of the core circadian clock genes Bmal1 and Per2, as well as in the DNA repair enzyme, MGMT. Independent measures of MGMT transcript levels and promoter methylation also showed daily rhythms intrinsic to GBM cells. These cells were more susceptible to TMZ when delivered at the daily peak of Bmal1 transcription. We found that in vivo morning administration of TMZ also decreased tumor size and increased body weight compared to evening drug delivery in mice bearing GBM xenografts. Finally, inhibition of MGMT activity with O6-Benzylguanine abrogated the daily rhythm in sensitivity to TMZ in vitro by increasing sensitivity at both the peak and trough of Bmal1 expression. CONCLUSION: We conclude that chemotherapy with TMZ can be dramatically enhanced by delivering at the daily maximum of tumor Bmal1 expression and minimum of MGMT activity and that scoring MGMT methylation status requires controlling for time of day of biopsy.

In situ metagenomics: A platform for rapid sequencing and analysis of metagenomes in less than one day.

We present here a complete system for metagenomic analysis that allows performing the sequencing and analysis of a medium-size metagenome in less than one day. This unprecedented development was possible due to the conjunction of state-of-the-art experimental and computational advances: a portable laboratory suitable for DNA extraction and sequencing with nanopore technology; the powerful metagenomic analysis pipeline SqueezeMeta, capable to provide a complete analysis in a few hours and using scarce computational resources; and tools for the automatic inspection of the results via a graphical user interface, that can be coupled to a web server to allow remote visualization of data (SQMtools and SQMxplore). We have tested the feasibility of our approach in the sequencing of the microbiota associated to volcanic rocks in La Palma, Canary Islands. Also, we did a two-day sampling campaign of marine waters in which the results obtained on the first day guided the experimental design of the second day. We demonstrate that it is possible to generate metagenomic information in less than one day, making it feasible to obtain taxonomic and functional profiles fast and efficiently, even in field conditions. This capacity can be used in the further to perform real-time functional and taxonomic monitoring of microbial communities in remote areas.

Linking the impact of bacteria on phytoplankton growth with microbial community composition and co-occurrence patterns.

The interactions between microalgae and bacteria have recently emerged as key control factors which might contribute to a better understanding on how phytoplankton communities assemble and respond to environmental disturbances. We analyzed partial 16S rRNA and 18S rRNA genes from a total of 42 antibiotic bioassays, where phytoplankton growth was assessed in the presence or absence of an active bacterial community. A significant negative impact of bacteria was observed in 18 bioassays, a significant positive impact was detected in 5 of the cases, and a non-detectable effect occurred in 19 bioassays. Thalasiossira spp., Chlorophytes, Vibrionaceae and Alteromonadales were relatively more abundant in the samples where a positive effect of bacteria was observed compared to those where a negative impact was observed. Phytoplankton diversity was lower when bacteria negatively affect their growth than when the effect was beneficial. The phytoplankton-bacteria co-occurrence subnetwork included many significant Chlorophyta-Alteromonadales and Bacillariophyceae-Alteromonadales positive associations. Phytoplankton-bacteria co-exclusions were not detected in the network, which contrasts with the negative effect of bacteria on phytoplankton growth frequently detected in the bioassays, suggesting strong competitive interactions. Overall, this study adds strong evidence supporting the key role of phytoplankton-bacteria interactions in the microbial communities.

[Use of screens, sedentary lifestyle, and physical activity in children under six years of age, during the period of preventive and compulsory social isolation in AMBA: online survey]. / Uso de pantallas, sedentarismo y actividad física en los niños menores de seis años, durante el periodo de aislamiento social preventivo y obligatorio en AMBA: encuesta en línea.

Introduction: Within the context of the COVID-19 pandemic, children were confined to their homes. The Buenos Aires Metropolitan Area (AMBA) region has the highest urban density in Argentina and has been the epicenter of COVID-19 infections and deaths. The objectives of this study are to characterize the habits of screen use, sedentary lifestyle and physical activity and describe their associations, in children under six years of age from AMBA, Argentina. Methods: analytical and cross-sectional study, in which data from the online survey (Google Forms®) were used. Through a non-probabilistic convenience sampling, fathers, mothers, legal guardians, and caregivers of children under six years of age were invited to participate. The survey was available during four months of the quarantine period and was made up of 31 questions about screen use, physical activity, and sedentary lifestyle of children. Results: 256 surveys were answered, the majority (79.6%) were female and 50.8% in the age range of 31-40 years. Conclusion: The television is the most used and children begin to use touch screens from one year. The play time of adults with children and the time that they usually carry out activities such as reading a book or drawing is one to two hours per day. There was an association between owning the screen and using it for more hours. The lower the income level, the more time adults play with their children.

Introducción: Dentro del contexto de la pandemia de COVID-19, los niños se quedaron confinados en sus hogares. La región del Área Metropolitana de Buenos Aires (AMBA) concentra la mayor densidad urbana de la Argentina y ha sido el epicentro de contagios y muertes por COVID-19. Los objetivos del presente estudio son caracterizar los hábitos del uso de pantalla, de sedentarismo y de la actividad física y describir sus asociaciones, en niños menores a seis años, del AMBA, Argentina. Métodos: estudio analítico y transversal, en que se utilizaron los datos de la encuesta en línea (Google Forms®). Mediante un muestreo no probabilístico por conveniencia, se invitaron a participar padres, madres, tutores legales y cuidadores de niños/as menores de seis años. La encuesta estuvo disponible durante cuatro meses del periodo de cuarentena y estuvo conformada por 31 preguntas sobre uso de pantalla, actividad física y sedentarismo de los niños. Resultados: fueron respondidas 256 encuestas, la mayoría (79,6%) era del género femenino y 50,8% en el rango etario de 31-40 años. Conclusión: La televisión es la más utilizada y los niños empiezan a usar pantallas táctiles a partir de un año. El tiempo de juego de los adultos con los niños y el tiempo que ellos suelen realizar actividades como leer un libro o dibujar es de una a dos horas por día. Hubo asociación entre ser dueño de la pantalla y usarla por más horas. Cuanto menor el nivel de ingreso, más tiempo los adultos juegan con sus niños.

Synergistic Effect of Brevetoxin BTX-3 and Ciguatoxin CTX3C in Human Voltage-Gated Nav1.6 Sodium Channels.

Emerging marine biotoxins such as ciguatoxins and brevetoxins have been widely and independently studied as food pollutants. Their maximum levels in food components were set without considering their possible synergistic effects as consequence of their coexistence in seafood and their action at the same cellular target. The absolute lack of data and regulations of the possible combined effects that both marine biotoxins may have raised the need to analyze their direct in vitro effects using electrophysiology techniques. The results presented in this study indicate that ciguatoxins and brevetoxins had a synergistic effect on human Nav1.6 voltage-gated sodium channels by hyperpolarizing their activation and inactivation states. The results presented here indicate that brevetoxin 3 (BTX-3) acts as partial agonist of human sodium channels, while ciguatoxin 3C (CTX3C) was a full agonist, explaining the differences in the effect of each toxin in the channel. Therefore, this work sets the cellular basis to further apply this type of studies to other food toxicants that may act synergistically and thus implement the corresponding regulatory limits considering their coexistence and the risks to human and animal health derived from it.