An inflection point in high-throughput proteomics with Orbitrap Astral: analysis of biofluids, cells, and tissues.

This technical note presents a comprehensive proteomics workflow for the new combination of Orbitrap and Astral mass analyzers across biofluids, cells, and tissues. Central to our workflow is the integration of Adaptive Focused Acoustics (AFA) technology for cells and tissue lysis, to ensure robust and reproducible sample preparation in a high-throughput manner. Furthermore, we automated the detergent-compatible single-pot, solid-phase-enhanced sample Preparation (SP3) method for protein digestion, a technique that streamlines the process by combining purification and digestion steps, thereby reducing sample loss and improving efficiency. The synergy of these advanced methodologies facilitates a robust and high-throughput approach for cells and tissue analysis, an important consideration in translational research. This work disseminates our platform workflow, analyzes the effectiveness, demonstrates reproducibility of the results, and highlights the potential of these technologies in biomarker discovery and disease pathology. For cells and tissues (heart, liver, lung, and intestine) proteomics analysis by data-independent acquisition mode, identifications exceeding 10,000 proteins can be achieved with a 24-minute active gradient. In 200ng injections of HeLa digest across multiple gradients, an average of more than 80% of proteins have a CV less than 20%, and a 45-minute run covers ~90% of the expressed proteome. In plasma samples including naive, depleted, perchloric acid precipitated, and Seer nanoparticle captured, all with a 24-minute gradient length, we identified 87, 108, 96 and 137 out of 216 FDA approved circulating protein biomarkers, respectively. This complete workflow allows for large swaths of the proteome to be identified and is compatible across diverse sample types.

Risk of endometrial cancer after RRSO in BRCA 1/2 carriers: a multicentre cohort study / Riesgo de cáncer de endometrio después de RRSO en portadoras de BRCA 1/2: un estudio de cohorte multicéntrico

Objective: To know the risk of endometrial cancer (EC) in a population of women with BRCA 1/2 pathogenic or likely pathogenic variants after risk-reducing salpingo-oophorectomy (RRSO). Methods: The study cohort included data from 857 women with BRCA mutations who underwent RRSO visited four hospitals in Catalonia, Spain, from January 1, 1999 to April 30, 2019. Standardized incidence ratio (SIR) of EC was calculated in these patients using data from a regional population-based cancer registry. Results: After RRSO, eight cases of EC were identified. Four in BRCA 1 carriers and four in BRCA2 carriers. The expected number of cases of EC was 3.67 cases, with a SIR of 2.18 and a 95% CI (0.93–3.95). Conclusions: In our cohort, the risk of EC in BRCA1/2 carriers after RRSO is not greater than expected. Hysterectomy is not routinely recommended for these patients.(AU)

Medical students' knowledge about human papillomavirus (HPV), HPV vaccine and head and neck cancer.

The Human Papillomavirus (HPV) is a sexually transmitted infection that significantly affects the population worldwide. HPV preventive methods include vaccination, prophylactics, and education. Different types of cancers associated with HPV usually take years or decades to develop after infections, such as Head and Neck Cancer(HNC). Therefore, HPV prevention can be considered cancer prevention. A sample of medical students in Puerto Rico was evaluated to assess their knowledge about HPV, HPV vaccine, and HNC through two previously validated online questionnaires composed of 38 dichotomized questions, we measured HPV, HPV vaccination(HPVK), and HNC knowledge (HNCK). Out of 104 students surveyed, the mean HPVK score obtained was 20.07/26, SD = 3.86, while the mean score for HNCK was 6.37/12, SD = 1.78. Bidirectional stepwise regression showed study year and HPV Vaccine name had been the most influential variables on HPVK and HNCK. MS1 participants scored lower than MS2-MS4 participants, with no significant difference between MS2-MS4 scores. The results reveal knowledge gaps in HPV/HPV Vaccine and HNC among surveyed medical students. Our findings also suggest an association between knowledge of personal vaccination status, self-perceived risk, and how uncertainty in these factors may affect the medical students' understanding of HPV, HPV vaccination, and associated cancers.

The Impact of Sibling Relationships on Behavioral and Sexual Health among Latino Sexual Minority Men.

Pre-exposure prophylaxis (PrEP) is a highly effective method for preventing HIV acquisition and plays a crucial role in the Ending the HIV Epidemic in the US initiative. However, there are various barriers that hinder the access and uptake of PrEP among Latino sexual minority men (SMM) at individual, interpersonal, and cultural levels. While the significance of cultural factors in designing and implementing HIV prevention programs for Latino populations has been consistently emphasized in the literature and prioritized by the Centers for Disease Control and Prevention, few studies have directly integrated these cultural factors into their programs. Our study aimed to investigate the potential influence of siblings in promoting the utilization of PrEP for HIV prevention, an area that has received limited attention. We conducted interviews with 31 pairs of siblings between December 2020 and January 2021, which were held in either English or Spanish and lasted approximately 45-60 min. The data were analyzed using a deductive thematic content analysis approach. The interviews revealed several key themes and categories, including: (1) Sibling support for coming out; (2) Types of support that siblings provide to each other for behavior change; (3) Sibling support for PrEP; and (4) The impact of the study interview on the quality of the sibling relationships. Our findings indicated that siblings were willing to provide support for PrEP in various ways, ranging from emotional support for brothers who may be concerned about potential rejection to practical support such as transportation or financial assistance. These results have significant implications for the design of HIV prevention interventions for Latinos. Incorporating siblings or other extended family members into these interventions can facilitate communication between siblings and their brothers, ultimately encouraging the use of PrEP or similar prevention methods. By considering the unique dynamics and support systems within Latino communities, researchers can develop more effective strategies to promote HIV prevention and support the well-being of Latino SMM.

Consensus Pharmacophore Strategy For Identifying Novel SARS-Cov-2 Mpro Inhibitors from Large Chemical Libraries.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main Protease (Mpro) is an enzyme that cleaves viral polyproteins translated from the viral genome and is critical for viral replication. Mpro is a target for anti-SARS-CoV-2 drug development, and multiple Mpro crystals complexed with competitive inhibitors have been reported. In this study, we aimed to develop an Mpro consensus pharmacophore as a tool to expand the search for inhibitors. We generated a consensus model by aligning and summarizing pharmacophoric points from 152 bioactive conformers of SARS-CoV-2 Mpro inhibitors. Validation against a library of conformers from a subset of ligands showed that our model retrieved poses that reproduced the crystal-binding mode in 77% of the cases. Using models derived from a consensus pharmacophore, we screened >340 million compounds. Pharmacophore-matching and chemoinformatics analyses identified new potential Mpro inhibitors. The candidate compounds were chemically dissimilar to the reference set, and among them, demonstrating the relevance of our model. We evaluated the effect of 16 candidates on Mpro enzymatic activity finding that seven have inhibitory activity. Three compounds (1, 4, and 5) had IC50 values in the midmicromolar range. The Mpro consensus pharmacophore reported herein can be used to identify compounds with improved activity and novel chemical scaffolds against Mpro. The method developed for its generation is provided as an open-access code (https://github.com/AngelRuizMoreno/ConcensusPharmacophore) and can be applied to other pharmacological targets.

Hypothermia and temperature modulation for intracerebral hemorrhage (ICH): pathophysiology and translational applications.

Background: Intracerebral hemorrhage (ICH) still poses a substantial challenge in clinical medicine because of the high morbidity and mortality rate that characterizes it. This review article expands into the complex pathophysiological processes underlying primary and secondary neuronal death following ICH. It explores the potential of therapeutic hypothermia as an intervention to mitigate these devastating effects. Methods: A comprehensive literature review to gather relevant studies published between 2000 and 2023. Discussion: Primary brain injury results from mechanical damage caused by the hematoma, leading to increased intracranial pressure and subsequent structural disruption. Secondary brain injury encompasses a cascade of events, including inflammation, oxidative stress, blood-brain barrier breakdown, cytotoxicity, and neuronal death. Initial surgical trials failed to demonstrate significant benefits, prompting a shift toward molecular mechanisms driving secondary brain injury as potential therapeutic targets. With promising preclinical outcomes, hypothermia has garnered attention, but clinical trials have yet to establish its definitive effectiveness. Localized hypothermia strategies are gaining interest due to their potential to minimize systemic complications and improve outcomes. Ongoing and forthcoming clinical trials seek to clarify the role of hypothermia in ICH management. Conclusion: Therapeutic hypothermia offers a potential avenue for intervention by targeting the secondary injury mechanisms. The ongoing pursuit of optimized cooling protocols, localized cooling strategies, and rigorous clinical trials is crucial to unlocking the potential of hypothermia as a therapeutic tool for managing ICH and improving patient outcomes.

Impact of natural events on metal bioaccumulation in Anemonia sulcata.

Samples of Anemonia sulcata were collected in 2022 from different areas of the Canary Islands affected by different natural contamination sources, such sandstorms, submarine volcanic activity, continuous rainfall, upwelling and dinoflagellate blooms. Significant differences were observed between the zones for the metals and trace elements analyzed (Al, Zn, Cd, Pb, Ni, Co, Fe, B, Cu, Mg and Li). Anemones from volcanic areas showed higher levels of Cd, Pb and Ni. Individuals from sandstorm areas showed elevated levels of Al, Zn and Fe. Samples collected from areas affected by upwelling processes had higher concentrations of Cu, Mg and Li. Finally, the areas affected by dinoflagellates showed lower levels of Zn, Pb, Fe, Mg and Li. The study reveals how natural phenomena dramatically influence metal accumulation in A. sulcata, which is of great value for anticipating and managing potential problems associated with public health.

Onyx®, A New Tool for Intraoperative Localization of Liver Lesions.

BACKGROUND: Precise preoperative localization of liver tumors facilitates successful surgical procedures, Intraoperative ultrasonography is a sensitive imaging modality. However, the presence of small non-palpable isoechoic intraparenchymal lesions may be challenging intraoperatively. METHODOLOGY AND MATERIAL DESCRIPTION: Onyx® is a non-adhesive liquid agent comprised of ethylene-vinyl alcohol usually used dissolved in dimethyl-sulfoxide and suspended micronized tantalum powder to provide contrast for visualization under fluoroscopy and ultrasonography and a macroscopic black shape. This embolization material has been increasingly used for the embolization of intracranial arteriovenous malformations. We present the novel application of Onyx® on liver surgery. CURRENT STATUS: We present the case of a female, 55 years-old, whose medical history revealed an elective sigmoidectomy (pT3N1a). After 17 months of follow up, by PET-CT scan, the patient was diagnosed of a small intraparenchymal hypo-attenuated 13 mm tumor located at segment V consistent with metachronous colorectal liver metastasis. Open metastasectomy was performed, ultrasonography-guided Onyx® infusion was delivered the day after, intraoperative ultrasonography showed a palpable hyperechoic material with a posterior acoustic shadowing artifact around the lesion. Onyx® is a promising new tool, without any previous application on liver surgery, feasible with advantages in small not palpable intraparenchymal liver lesions.

Comparative analysis of metal pollution in the Atlantic Ocean and Mediterranean Sea: insights from Anemonia sulcata study.

In this study, we investigated metal concentrations in Anemonia sulcata specimens from various locations in the Atlantic Ocean and the Mediterranean Sea. A total of 84 individuals were sampled from specific zones, and their tissue samples were processed for metal analysis using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES). The results revealed notable differences in metal concentrations among the studied regions. The Atlantic Ocean and the Mediterranean Sea were found to have distinct patterns of marine pollution, influenced by a complex interplay of geographical, demographic, industrial, and environmental policy factors. Conversely, the semi-enclosed Mediterranean Sea has a lower natural dilution capacity, leading to the accumulation and prolonged presence of pollutants. Population density and industrial activities in coastal areas play a significant role in pollution disparities between the Mediterranean Sea and the Atlantic Ocean. The Mediterranean coasts, with higher population densities and intensive industrial operations, experience greater strain on marine ecosystems due to increased pollution sources. Additionally, environmental policies and management approaches differ between the two regions, contributing to variations in pollution response and regulation.

A novel study, textbook outcome in adrenalectomy: retrospective observational study in an endocrine surgical unit.

Textbook outcome is a multidimensional quality management tool that uses a set of traditional surgical measures to reflect an "ideal" surgical result for a particular pathology. Retrospective study of all patients undergoing scheduled for adrenal tumors surgery at an endocrine surgery unit from January 2010-December 2022. The definition of Textbook Outcome were: R0 resection, no Clavien-Dindo ≥ IIIa complications, no prolonged stay (< P75), no readmissions, and no mortality in the first 30 days. The main objective was to analyze the rate of Textbook Outcome obtained. One hundred and five patients were included in the study. Textbook Outcome was achieved in 71.4%. Surgical approach (p < 0.001), Charlson scale (p = 0.031), American Society of Anesthesiologists Classification (p = 0.047) and surgical time (p < 0.001) were all significantly associated with the achievement of Textbook Outcome. The laparoscopic approach was associated as an independent factor with obtaining Textbook Outcome (OR:5.394; p = 0.016), as was surgical time (OR:0.986; p = 0.004). Textbook Outcome is a novel, useful, easy-to-interpret tool for measuring results in adrenal surgery. The laparoscopic approach is associated with a higher rate of "ideal" surgical results. The study was registered in the public accessible database clinicaltrials.gov with the ClinicalTrials.gov ID: NCT05888753.

OGT prevents DNA demethylation and suppresses the expression of transposable elements in heterochromatin by restraining TET activity genome-wide.

The O- GlcNAc transferase OGT interacts robustly with all three mammalian TET methylcytosine dioxygenases. We show here that deletion of the Ogt gene in mouse embryonic stem cells (mESC) results in a widespread increase in the TET product 5-hydroxymethylcytosine (5hmC) in both euchromatic and heterochromatic compartments, with concomitant reduction of the TET substrate 5-methylcytosine (5mC) at the same genomic regions. mESC engineered to abolish the TET1-OGT interaction likewise displayed a genome-wide decrease of 5mC. DNA hypomethylation in OGT-deficient cells was accompanied by de-repression of transposable elements (TEs) predominantly located in heterochromatin, and this increase in TE expression was sometimes accompanied by increased cis -expression of genes and exons located 3' of the expressed TE. Thus, the TET-OGT interaction prevents DNA demethylation and TE expression in heterochromatin by restraining TET activity genome-wide. We suggest that OGT protects the genome against DNA hypomethylation and impaired heterochromatin integrity, preventing the aberrant increase in TE expression observed in cancer, autoimmune-inflammatory diseases, cellular senescence and ageing.

Specific targeting of the NRF2/ß-TrCP axis promotes beneficial effects in NASH.

Non-alcoholic steatohepatitis (NASH) is a common chronic liver disease that compromises liver function, for which there is not a specifically approved medicine. Recent research has identified transcription factor NRF2 as a potential therapeutic target. However, current NRF2 activators, designed to inhibit its repressor KEAP1, exhibit unwanted side effects. Alternatively, we previously introduced PHAR, a protein-protein interaction inhibitor of NRF2/ß-TrCP, which induces a mild NRF2 activation and selectively activates NRF2 in the liver, close to normal physiological levels. Herein, we assessed the effect of PHAR in protection against NASH and its progression to fibrosis. We conducted experiments to demonstrate that PHAR effectively activated NRF2 in hepatocytes, Kupffer cells, and stellate cells. Then, we used the STAM mouse model of NASH, based on partial damage of endocrine pancreas and insulin secretion impairment, followed by a high fat diet. Non-invasive analysis using MRI revealed that PHAR protects against liver fat accumulation. Moreover, PHAR attenuated key markers of NASH progression, including liver steatosis, hepatocellular ballooning, inflammation, and fibrosis. Notably, transcriptomic data indicate that PHAR led to upregulation of 3 anti-fibrotic genes (Plg, Serpina1a, and Bmp7) and downregulation of 6 pro-fibrotic (including Acta2 and Col3a1), 11 extracellular matrix remodeling, and 8 inflammatory genes. Overall, our study suggests that the mild activation of NRF2 via the protein-protein interaction inhibitor PHAR holds promise as a strategy for addressing NASH and its progression to liver fibrosis.

Metal content in Sardina pilchardus during the period 2014-2022 in the Canary Islands (Atlantic EC, Spain).

The contamination present in an organism varies depending on biological and oceanographic conditions, so monitoring the same species is of great importance to understand the state of the ecosystem. Fifteen specimens in Sardina pilchardus between 12 and 15 cm in total length were collected during the second half of January of each of the study years (2014, 2016, 2018, 2020 and 2022). Samples were analyzed with Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) to measure metals (Al, Cd, Cu, Fe, Li, Ni, Pb and Zn) in mg/Kg. There was a progressive decrease in Pb content over the period, with the highest concentration being obtained in 2014 (0.086 ± 0.065 mg/kg). Locally important oceanic-atmospheric events may occur in the study period that strongly impact the tissue composition of marine organisms. In this case, discontinuous trends were evident in some of the metal concentrations analyzed in the muscle of European sardine in the Canary Islands.

Host genetic regulation of human gut microbial structural variation.

Although the impact of host genetics on gut microbial diversity and the abundance of specific taxa is well established1-6, little is known about how host genetics regulates the genetic diversity of gut microorganisms. Here we conducted a meta-analysis of associations between human genetic variation and gut microbial structural variation in 9,015 individuals from four Dutch cohorts. Strikingly, the presence rate of a structural variation segment in Faecalibacterium prausnitzii that harbours an N-acetylgalactosamine (GalNAc) utilization gene cluster is higher in individuals who secrete the type A oligosaccharide antigen terminating in GalNAc, a feature that is jointly determined by human ABO and FUT2 genotypes, and we could replicate this association in a Tanzanian cohort. In vitro experiments demonstrated that GalNAc can be used as the sole carbohydrate source for F. prausnitzii strains that carry the GalNAc-metabolizing pathway. Further in silico and in vitro studies demonstrated that other ABO-associated species can also utilize GalNAc, particularly Collinsella aerofaciens. The GalNAc utilization genes are also associated with the host's cardiometabolic health, particularly in individuals with mucosal A-antigen. Together, the findings of our study demonstrate that genetic associations across the human genome and bacterial metagenome can provide functional insights into the reciprocal host-microbiome relationship.

Risk of endometrial cancer after RRSO in BRCA 1/2 carriers: a multicentre cohort study.

OBJECTIVE: To know the risk of endometrial cancer (EC) in a population of women with BRCA 1/2 pathogenic or likely pathogenic variants after risk-reducing salpingo-oophorectomy (RRSO). METHODS: The study cohort included data from 857 women with BRCA mutations who underwent RRSO visited four hospitals in Catalonia, Spain, from January 1, 1999 to April 30, 2019. Standardized incidence ratio (SIR) of EC was calculated in these patients using data from a regional population-based cancer registry. RESULTS: After RRSO, eight cases of EC were identified. Four in BRCA 1 carriers and four in BRCA2 carriers. The expected number of cases of EC was 3.67 cases, with a SIR of 2.18 and a 95% CI (0.93-3.95). CONCLUSIONS: In our cohort, the risk of EC in BRCA1/2 carriers after RRSO is not greater than expected. Hysterectomy is not routinely recommended for these patients.

Flow and temporal effects on the sonolytic defluorination of perfluorooctane sulfonic acid.

The removal of per- and polyfluoroalkyl substance (PFAS) pollution from the environment is a globally pressing issue, due to some PFAS' recalcitrant, bioaccumulative, and carcinogenic nature. Destruction via ultrasonic waves (sonolysis) is a promising contender for industrialisation due to; moderate power consumption, applicability to several PFAS and sample types, and limited by-products. Liquid flow rate through an ultrasonic reactor can affect the size, shape, and spatial distribution of ultrasonic cavities and hence their chemical activity. Such effects have not been studied during PFAS sonolysis, and temporal effects have not been studied much beyond the reactant concentration. Here, the effects of varying recirculating flow rate on the ultrasonic defluorination of perfluorooctane sulfonic acid (PFOS) and implications for industrial scale up are presented. Under the ultrasonic power (200 W L-1, 2.27 W cm-2) and frequency (410 kHz) used, flow rates of 79 and 214 ml min-1 enhanced defluorination up to 14 % during 30 min of treatment. However, these effects were temporal and most significant in the initial minutes of treatment. This indicated a dynamic bubble size distribution which stabilised after around 15 min. Defluorination rates of PFOS were compared with measured potassium iodide dosimetry, calorimetry, sonoluminescence (SL), and sonochemiluminescence (SCL). Flow rates which enhanced defluorination correlated moderately with enhanced SCL and negatively impacted SL, calorimetry, and dosimetry. Effects were attributed to perturbed cavity surfaces, leading to asymmetric cavity collapse, and the possibility of enhanced solvated electron production/interaction. SL, SCL, dosimetry, and calorimetric measurements were also temporal, and each showed different times to equilibrate. Flow rates of 439 and 889 ml min-1 returned all sonochemical measurements to the levels without flow, likely due to continued collapse temperature quenching by furthered bubble asymmetry. Flow also enhanced reactor cooling, which is significant for industrial temperature control. The pump energy consumed was small (≈1.9 %) compared to that of the amplifier and chiller, hence, PFOS defluorination was more cost-effective using flow. However, the effect may be limited for the longer treatment times needed for environmental remediation.

A Comparative Analysis of Sparisoma cretense in Island Environments: Unraveling Metal Accumulation Differences in the Canary Islands (Spain, NW African Waters).

This study investigates the impact of varying environmental conditions on the metal composition within the tissues of Sparisoma cretense, contributing to the understanding necessary to offer scientifically sound advice regarding the health status of this species. This knowledge extends beyond fishery production, encompassing implications for food security. The data span the years 2022 and 2023, encompassing both cold and warm climatic seasons. The concentrations of various metals, such as Al, Zn, Cd, Pb, Fe, and Cu, exhibited noteworthy variations across the islands, with significant increases recorded in 2023, particularly during the warm season. The intricate interplay between multiple factors shaped the availability of the analyzed elements in S. cretense. Factors such as rising temperatures during the warm season increased biological activity in marine ecosystems, seasonal fluctuations in weather conditions, water quality, and anthropogenic influences, all contributing to the observed variations in metal concentrations. Additionally, the geological composition of each island and the patterns of marine currents and sediment transport play pivotal roles in these differences. Comprehensive scientific research, monitoring, and environmental surveillance are essential for a holistic understanding of this variability and providing valuable insights for the conservation and management of marine ecosystems in the Canary archipelago.

Soft Drinks as a Dietary Source of Fluoride Exposure.

High fluoride exposures can lead to adverse effects such as dental and bone fluorosis, as well as endocrine and cognitive developmental problems. Water is the main dietary source of this ion, although significant concentrations have also been detected in other beverages widely consumed by the population such as soft drinks. A total of 200 soft drink samples (60 flavoured, 70 extracts, 60 fruit juice and 10 soft drinks) were analysed by fluoride ion selective potentiometry. A consumption of 330 mL was estimated for exposure assessment and subsequent F-risk assessment by soft drink consumption. The highest average concentration was found in extract soft drinks (2.45 ± 1.15 mg/L), followed by flavoured (1.71 ± 2.29 mg/L) and carbonated soft drinks (1.38 ± 0.40 mg/L), while the lowest was found in fruit juice soft drinks (1.09 ± 0.62 mg/L). The flavours with the highest concentration were tea-melon and tea-passion fruit with 3.66 ± 0.40 and 3.17 ± 0.56 mg/L respectively and the lowest was lemon flavour with 0.69 mg/L. The contribution of these beverages, considering the UL (Upper level) reference values set by EFSA (European Food Safety Authority) are between 3.28-41.78%, depending on age group and sex.

Impact of the lockdown period during the COVID-19 pandemic on the metal content of the anemone Anemonia sulcata in the Canary Islands (CE Atlantic, Spain).

Anemones, specifically the species Anemonia sulcata, are cnidarians that serve as bioindicators in marine ecosystems, indicating the health of the environment and changes in environmental conditions. Monitoring anemone populations and studying their well-being and distribution provide valuable insights into marine ecosystem conditions. This study aimed to investigate the impact of the SARS-CoV-2 pandemic on the metal content of Anemonia sulcata. Over a six-year period (2017-2022), twenty specimens of Anemonia sulcata were collected in Tenerife, Spain. The results showed that in 2020, during the two-month lockdown in Spain from March to May when tourism was halted, A. sulcata exhibited the lowest concentrations of various metals studied (Al, Cd, Cu, Fe, Pb, and Zn). This finding suggests that the reduced anthropogenic pressure on the coast due to the absence of tourism significantly decreased pollution levels. Therefore, the study emphasizes the importance of promoting sustainable tourism worldwide. The research highlights that minimizing human impact on coastal areas through responsible tourism practices can effectively reduce pollution in marine ecosystems.