Improvement in edema and cognitive recovery after moderate traumatic brain injury with the neurosteroid prodrug NTS-104.

Neuroactive steroids reduce mortality, decrease edema, and improve functional outcomes in preclinical and clinical traumatic brain injury (TBI) studies. In this study, we tested the efficacy of two related novel neuroactive steroids, NTS-104 and NTS-105, in a rat model of TBI. NTS-104 is a water-soluble prodrug of NTS-105, a partial progesterone receptor agonist. To investigate the effects of NTS-104 on TBI recovery, adult male Sprague Dawley rats received moderate parasagittal fluid-percussion injury or sham surgery and were treated with vehicle or NTS-104 (10 â€‹mg/kg, intramuscularly) at 4, 10, 24, and 48 â€‹h post-TBI. The therapeutic time window was also assessed using the neuroactive steroid NTS-105 (3 â€‹mg/kg, intramuscularly). Edema in the parietal cortex and hippocampus, measured at 3 days post-injury (DPI), was reduced by NTS-104 and NTS-105. NTS-105 was effective in reducing edema when given at 4, 10, or 24 â€‹h post-injury. Sensorimotor deficits in the cylinder test at 3 DPI were ameliorated by NTS-104 and NTS-105 treatment. Cognitive recovery, assessed with cue and contextual fear conditioning and retention of the water maze task assessed subacutely 1-3 weeks post-injury, also improved with NTS-104 treatment. Cortical and hippocampal atrophy at 22 DPI did not improve, indicating that NTS-104/NTS-105 may promote post-TBI cognitive recovery by controlling edema and other processes. These results demonstrate that NTS-104/NTS-105 is a promising therapeutic approach to improve motor and cognitive recovery after moderate TBI.

The epithelial barrier theory and its associated diseases.

The prevalence of many chronic noncommunicable diseases has been steadily rising over the past six decades. During this time, over 350,000 new chemical substances have been introduced to the lives of humans. In recent years, the epithelial barrier theory came to light explaining the growing prevalence and exacerbations of these diseases worldwide. It attributes their onset to a functionally impaired epithelial barrier triggered by the toxicity of the exposed substances, associated with microbial dysbiosis, immune system activation, and inflammation. Diseases encompassed by the epithelial barrier theory share common features such as an increased prevalence after the 1960s or 2000s that cannot (solely) be accounted for by the emergence of improved diagnostic methods. Other common traits include epithelial barrier defects, microbial dysbiosis with loss of commensals and colonization of opportunistic pathogens, and circulating inflammatory cells and cytokines. In addition, practically unrelated diseases that fulfill these criteria have started to emerge as multimorbidities during the last decades. Here, we provide a comprehensive overview of diseases encompassed by the epithelial barrier theory and discuss evidence and similarities for their epidemiology, genetic susceptibility, epithelial barrier dysfunction, microbial dysbiosis, and tissue inflammation.

Intranasal versus oral treatments for allergic rhinitis: A systematic review with meta-analysis.

BACKGROUND: Treatments for allergic rhinitis include intranasal or oral medications. OBJECTIVE: To perform a systematic review with meta-analysis comparing the effectiveness of intranasal corticosteroids or antihistamines versus oral antihistamines or leukotriene receptor antagonists in improving allergic rhinitis symptoms and quality of life. METHODS: We searched four bibliographic databases and three clinical trial datasets for randomised controlled trials (i) assessing patients ≥12 years old with seasonal or perennial allergic rhinitis, and (ii) comparing intranasal corticosteroids or antihistamines versus oral antihistamines or leukotriene receptor antagonists. We performed a meta-analysis of the Total Nasal Symptom Score (TNSS), Total Ocular Symptom Score (TOSS), Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ), development of adverse events, and withdrawals due to adverse events. Certainty of evidence was assessed using GRADE. RESULTS: We included 35 studies, most of which assessed patients with seasonal allergic rhinitis and displayed an unclear risk of bias. Superiority of intranasal treatments was found for all assessed outcomes. Intranasal corticosteroids were more effective than oral antihistamines at improving the TNSS (MD=-0.86; 95%CI=-1.21;-0.51; I2=70%), TOSS (MD=-0.36; 95%CI=-0.56;-0.17; I2=0%) and RQLQ (MD=-0.88; 95%CI=-1.15;-0.61; I2=0%), being mostly associated with clinically meaningful improvements. Superiority of intranasal corticosteroids at improving the TNSS was also found against oral leukotriene receptor antagonists (MD=-1.05; 95%CI=-1.33;-0.77). Intranasal antihistamines were more effective than oral antihistamines at improving the TNSS (MD=-0.47; 95%CI=-0.81;-0.14; I2=0%) and RQLQ (MD=-0.31; 95%CI=-0.56;-0.06; I2=0%). CONCLUSIONS: Randomized controlled trials suggest that intranasal treatments are more effective than oral treatments at improving symptoms and quality of life in seasonal allergic rhinitis.

Analysis of care and gender stereotypes in nursing scientific research: a scoping review.

OBJECTIVE: To map evidence about care and gender stereotypes in nursing scientific research. METHOD: A scoping review developed under the JBI framework with analysis of gender perspective in care approaches. The searches were carried out on January 31, 2023 in SciELO, Scopus, CINAHL, PubMed, BDENF. RESULTS: Of the 3,743 studies located, 25 were included. Evidence was grouped into categories: essentially female care (n = 9; 36%); calling and service of love (n = 3; 12%); erasure of gender inequalities (n = 2; 8%); "inadequate and harmful" care (n = 5; 20%); neutralization of gender and bodies (n = 3; 12%); and reporting oppression in care work (n = 3; 12%). CONCLUSION: Most scientific research on care reproduces gender stereotypes that reinforce the oppression of women in nursing. In contrast, resistance denounces naturalization of care as "inadequate and harmful", for perpetuating gender oppression in care work.

Integrating nutrition and culinary medicine into preclinical medical training.

BACKGROUND: Poor nutrition is a leading cause of preventable death, but is inconsistently taught in medical education and inadequately discussed in medical care. To overcome this problem, we developed a hybrid nutrition team-based learning/culinary medicine approach to integrate practical nutrition knowledge and basic cooking skills into the training of future health professionals. METHODS: Nutrition was integrated into the systems-based courses at a college of osteopathic medicine, complemented by culinary medicine sessions based on the Health meets Food curriculum (HmF; culinarymedicine.org). Students participated in the program for one year and two cohorts of students were included in this analysis. Outcomes were measured via online food frequency questionnaire (FFQ, Vioscreen, Viocare, Inc) and surveys administered via Qualtrics online survey software. Diet quality was measured using the Healthy Eating Index (HEI)-2015. Data were analyzed using SAS 9.4. RESULTS: One hundred and ninety-five first year students completed a baseline FFQ (97.5% response rate). Mean age of students was 26 years, 47% were female (n = 92/195). The average BMI of participants was 24.8 kg/m2 (range 17-45.4) and the majority of participants reported being active. Seventy-five students (38%) completed an end of year FFQ. Diet quality was poor among students at baseline (n = 195; 67.59 (SD 10.54)) and improved slightly but significantly at the end of year 1 (n = 75, 69.63 (SD: 12.42), p = 0.04). The survey was administered to the second cohort only; 63 students responded (53% response rate). Talking to patients about nutrition was seen as more relevant to future practice among respondents than talking to patients about safe sex, weight, tobacco, alcohol, other substance abuse and domestic violence. CONCLUSIONS: This study evaluated the nutrition and culinary medicine curriculum at a new college of osteopathic medicine. Students rated the program highly and attendance was excellent, even though not required. Student diet quality did not decline over the first year of medical school. Students rated talking to patients about nutrition as highly relevant, providing encouragement that they will do so in future practice. We believe our work shows that nutrition can be integrated into the training of future physicians and that it may pay dividends, particularly with the increasing awareness of the importance of preventive care.

Phosphate-binding proteins and peptides: from molecular mechanisms to potential applications.

Selective binding of phosphate is vital to multiple aims including phosphate transport into cells and phosphate-targeted applications such as adsorption-based water treatment and sensing. High-affinity phosphate-binding proteins and peptides offer a nature-inspired means of efficiently binding and separating phosphate from complex matrices. The binding protein PstS is characterized by a Venus flytrap topology that confers exceptional phosphate affinity and selectivity, and is effective even at low phosphate concentrations, all of which are essential for applications such as phosphate sensing, removal, and recovery. The binding event is reversible under controlled conditions, making it germane to catch-and-release objectives that advance phosphorus sustainability. Peptides such as the P loop motif are also promising for such applications. Future advances in protein/peptide design can contribute to increased implementation in engineered systems.

Evaluation of carrageenans extracted by an eco-friendly technology as source for gelled matrices with potential food application.

Red macroalgae are considered an immense source of hydrocolloids (agar and carrageenan) that are gaining momentum in the food industry as an alternative to animal-based ones, like gelatin. This work evaluates carrageenans extracted from four different red macroalgae (Chondrus crispus, Mastocarpus stellatus, Sarcopeltis skottsbergii and Gigartina pistillata) by an eco-friendly process (hydrothermal extraction), for their possible employment as food additives considering purity requirements stated by the European Regulation. In general, carrageenans presented a suitable composition, although some sample presented lower sulfate content than 15 % and higher As content than 3 mg/kg, being only carrageenans from Chondrus crispus and Sarcopeltis skottsbergii appropriate for gelled matrices formulation. Different concentrations of hydrocolloids (1-5 %) and salts (0.1-1 M NaCl, CaCl2 and KCl) were evaluated to reach a desired consistency. Rheological behavior of said gels revealed a gel-like behavior, with G' > G" and practically frequency independency of the parameters. Overall, gels formulated with KCl achieved higher G' with maximum values of 100-1000 Pa, whereas the commercial gelled dessert (used as control) only achieved values of around 10 Pa. After 3 months of cold storage, all gels exhibited a strengthening of the gelled matrix, without water syneresis. The colorimetric parameters were also evaluated, showing higher inclination for red and yellow tones with modest lightness values (around 60 %). In this work, hydrothermally extracted carrageenans from Chondrus crispus and Sarcopeltis skottsbergii were assessed, laying the groundwork for further studies in this area.

In vivo dendritic cell reprogramming for cancer immunotherapy.

Immunotherapy can lead to long-term survival for some cancer patients, yet generalized success has been hampered by insufficient antigen presentation and exclusion of immunogenic cells from the tumor microenvironment. Here, we developed an approach to reprogram tumor cells in vivo by adenoviral delivery of the transcription factors PU.1, IRF8, and BATF3, which enabled them to present antigens as type 1 conventional dendritic cells. Reprogrammed tumor cells remodeled their tumor microenvironment, recruited, and expanded polyclonal cytotoxic T cells, induced tumor regressions, and established long-term systemic immunity in multiple mouse melanoma models. In human tumor spheroids and xenografts, reprogramming to immunogenic dendritic-like cells progressed independently of immunosuppression, which usually limits immunotherapy. Our study paves the way for human clinical trials of in vivo immune cell reprogramming for cancer immunotherapy.

The First Patient with Tibial Hemimelia-Polysyndactyly-Triphalangeal Thumb Syndrome Caused by De Novo c.423+4916 T>C ZRS Variant: A Case Report.

Genetic variants in the zone of polarizing activity regulatory sequence (ZRS) that induce ectopic expression of the SHH gene have been associated with different ZRS-related phenotypes. We report the first patient with a de novo variant, c.423+4916 T>C, in ZRS (previously classified as a variant of uncertain significance) that causes tibial hemimelia-polysyndactyly-triphalangeal thumb syndrome (THPTTS). A two-month-old male patient presented with bilateral preaxial polydactyly, triphalangeal thumb, and tibial agenesis and was heterozygous for the variant c.423+4916T>C (neither of his parents was a carrier). The findings obtained from the family study were sufficient to reclassify the variant from "uncertain significance" to "likely pathogenic" according to three criteria from the American College of Medical Genetics and Genomics guidelines, as follows: (1) absence of gnomAD, (2) confirmation of paternity and maternity, and (3) strong phenotype-genotype association. In ZRS-associated syndromes, a wide clinical spectrum has been observed, ranging from polydactyly to THPTTS; our patient has the most severe and rare phenotype. We did not perform functional assays. However, the c.423+4916T>C variant is flanked by three variants, which have been proven not only to cause the phenotype but also to increase the expression of SHH. Through all this data gathering, we consider the c.423+4916T>C variant to be causative of THPTTS.

Non-invasive assessment of severe liver fibrosis in patients with Fontan-associated liver disease: The VALDIG-EASL FONLIVER cohort.

BACKGROUND & AIMS: Fontan-type surgery is used as a palliation for congenital heart disease with univentricular physiology but may, in the long term, lead to advanced chronic liver disease. This study assessed the accuracy of conventional non-invasive models in assessing liver fibrosis and introduces a new risk score employing non-invasive tools. METHODS: A prospective, cross-sectional, observational study was conducted across five European centers and encompassing all consecutive adult patients with Fontan circulation, liver biopsy and non-invasive tests (elastography, APRI, FIB-4, Fibrosis score, Doha, GUCI, and AAR). The primary outcome was the identification of severe liver fibrosis on biopsy. Multivariable logistic regression identified non-invasive predictors of severe fibrosis, leading to the development and internal validation of a new scoring model named the FonLiver risk score. RESULTS: In total, 217 patients (mean [standard deviation] age, 27.9 [8.9] years; 50.7% males) were included. Severe liver fibrosis was present in 47.9% (95% CI 41.2%-54.5%) and correlated with a lower functional class, protein-losing enteropathy, and compromised cardiopulmonary and systemic hemodynamics. The final FonLiver risk score incorporated liver stiffness measurement using transient elastography and platelet count and demonstrated strong discrimination and calibration (area under the receiver operating curve [AUROC] of 0.81). The FonLiver risk score outperformed conventional prediction models (APRI, FIB-4, Fibrosis score, Doha, GUCI, and AAR), which all exhibited worse performance in our cohort (AUROC < 0.70 for all). CONCLUSION: Severe liver fibrosis is prevalent in adults following Fontan-type palliation and can be effectively estimated using with the novel FonLiver risk score. This scoring system can be easily incorporated into the routine assessment of patients with Fontan circulation.

Development of a new reference material for accurate measurements of lithium in Li-clays.

Currently, there are no national and international certified reference materials (CRM) in lithium clays that can make reliable and traceable lithium measurements for the International System of Units (SI). Hence, it is necessary to have references to meet the needs in terms of mining and activities that involve the use of lithium to favor the economy derived from its multiple uses and associated benefits in the exploration, exploitation, and handling of lithium ore. In this study, a candidate for reference material (RM) of Li in clays was developed and certified based on the provisions of ISO 17034:2016 and ISO Guide 35:2017. Different mass sizes of the RM (0.05, 0.1, and 0.25 g) were used to evaluate homogeneity. An isochronous study (short-term stability) was carried out in the assessment of stability, influenced by the effects of transport at different temperatures (20, 40, and 50 °C) for a determined time of 6 weeks, in addition to a classic (long-term) study for 19 weeks. The sample was treated using microwave-assisted acid digestion and Li measurements were performed using the analytical technique of Flame Atomic Absorption Spectrometry (FAAS). The CRM is homogeneous for the sample mass sizes of 0.05 and 0.1 g, and the mass fraction of w(Li) was stable in the RM for temperatures of 20, 40, and 50 °C. The determined period of validity was 3 years.

Advances in the approaches used to repurpose drugs for neuroblastoma.

INTRODUCTION: Neuroblastoma (NB) remains a challenging pediatric malignancy with limited treatment options, particularly for high-risk cases. Drug repurposing offers a convenient and cost-effective strategy for treating rare diseases like NB. Using existing drugs with known safety profiles accelerates the availability of new treatments, reduces development costs, and mitigates risks, offering hope for improved patient outcomes in challenging conditions. AREAS COVERED: This review provides an overview of the advances in approaches used to repurpose drugs for NB therapy. The authors discuss strategies employed in drug repurposing, including computational and experimental methods, and rational drug design, highlighting key examples of repurposed drugs with promising clinical results. Additionally, the authors examine the challenges and opportunities associated with drug repurposing in NB and discuss future directions and potential areas for further research. EXPERT OPINION: The fact that only one new drug has been approved in the last 30 years for the treatment of neuroblastoma plus a significant proportion of high-risk NB patients that remain uncurable, evidences the need for new fast and cost-effective alternatives. Drug repurposing may accelerate the treatment development process while reducing expenses and risks. This approach can swiftly bring effective NB therapies to market, enhancing survival rates and patient quality of life.

Implementing a Diet Risk Score (DRS) for Spanish-Speaking Adults in a Clinical Setting: A Feasibility Study.

Tools to briefly assess diet among US Spanish-speaking adults are needed to identify individuals at risk for cardiometabolic disease (CMD) related to diet. Two registered dietitian nutritionists (RDNs) recruited bilingual medical students to translate the validated Diet Risk Score (DRS) into Spanish (DRS-S). Participants were recruited from a federally qualified health center. Students administered the DRS-S and one 24-h recall (Automated Self-Administered 24-Hour (ASA24®) Dietary Assessment Tool) on one day; a second recall was administered within 1 week. Recalls were scored using the Healthy Eating Index (HEI)-2015, a measure of adherence to the Dietary Guidelines for Americans. Spearman correlations, weighted kappa, and ANOVA were conducted using SAS 9.4 to assess the relative validity of the DRS-S. Thirty-one Spanish-speaking adults (female: n = 17, 53%; mean age: 58 (42-69)) completed assessments. The mean DRS-S was 9 (SD = 4.2) (max: 27; higher score = higher risk) and the mean HEI-2015 score was 65.7 (SD = 9.7) (max: 100; higher score = lower risk), with significant agreement between measures (r: -0.45 (p = 0.01)), weighted kappa: -0.3 (p = 0.03). The DRS-S can be used in resource-constrained settings to assess diet for intervention and referral to RDNs. The DRS-S should be tested in clinical care to assess the impact of dietary changes to reduce CMD risk.

Selective human milk oligosaccharide utilization by members of the Bifidobacterium pseudocatenulatum taxon.

Human milk oligosaccharides (HMOs) are essentially unaffected by the digestive enzymes of the nursling and are known for their ability to enrich certain microbial species in the infant gut microbiota, in particular bifidobacteria. HMO metabolism has been studied in various bifidobacterial species such as B. breve, B. bifidum, and B. longum subsp. infantis. In the current study, we describe differential growth abilities elicited by twenty-three newly isolated Bifidobacterium pseudocatenulatum strains on particular HMOs, such as 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), lacto-N-tetraose (LNT), and lacto-N-neotetraose (LNnT). Through gene-trait matching and comparative genome analysis, we identified genes involved in the degradation of fucosylated HMOs in this strain set, while we employed a transcriptomic approach to facilitate the identification and characterization of genes and associated enzymes involved in LNT metabolism by strain B. pseudocatenulatum MM0196. A total of 252 publicly available genomes of the B. pseudocatenulatum taxon were screened for homologs of the glycosyl hydrolases (GHs) identified here as being required for selected HMO metabolism. From this analysis, it is clear that all members of this species possess homologs of the genes involved in LNT degradation, while genes required for degradation of fucosylated HMOs are variably present.IMPORTANCEOur findings allow a better understanding of the complex interaction between Bifidobacterium and its host and provide a roadmap toward future applications of B. pseudocatenulatum as a probiotic with a focus on infant health. Furthermore, our investigations have generated information on the role of HMOs in shaping the infant gut microbiota, thus also facilitating applications of HMOs in infant nutrition, with potential extension into the mature or adult gut microbiota. Supplementation of HMOs is known to result in the modulation of bacterial communities toward a higher relative abundance of bifidobacteria, which in turn enforces their ability to modulate particular immune functions and strengthen the intestinal barrier. This work may therefore inspire future studies to improve the formulation of neonatal nutritional products, aimed at facilitating the development of a healthy digestive and immune system and reducing the differences in gut microbiota composition observed between breastfed and formula-fed babies or full-term and preterm infants.

Automated High-Throughput Biological Sex Identification from Archeological Human Dental Enamel Using Targeted Proteomics.

Biological sex is key information for archeological and forensic studies, which can be determined by proteomics. However, the lack of a standardized approach for fast and accurate sex identification currently limits the reach of proteomics applications. Here, we introduce a streamlined mass spectrometry (MS)-based workflow for the determination of biological sex using human dental enamel. Our approach builds on a minimally invasive sampling strategy by acid etching, a rapid online liquid chromatography (LC) gradient coupled to a high-resolution parallel reaction monitoring (PRM) assay allowing for a throughput of 200 samples per day (SPD) with high quantitative performance enabling confident identification of both males and females. Additionally, we developed a streamlined data analysis pipeline and integrated it into a Shiny interface for ease of use. The method was first developed and optimized using modern teeth and then validated in an independent set of deciduous teeth of known sex. Finally, the assay was successfully applied to archeological material, enabling the analysis of over 300 individuals. We demonstrate unprecedented performance and scalability, speeding up MS analysis by 10-fold compared to conventional proteomics-based sex identification methods. This work paves the way for large-scale archeological or forensic studies enabling the investigation of entire populations rather than focusing on individual high-profile specimens. Data are available via ProteomeXchange with the identifier PXD049326.

Whole Exome Sequencing Identifies Epithelial and Immune Dysfunction-Related Biomarkers in Food Protein-Induced Enterocolitis Syndrome.

BACKGROUND: Food protein-induced enterocolitis syndrome (FPIES) is a food allergy primarily affecting infants, often leading to vomiting and shock. Due to its poorly understood pathophysiology and lack of specific biomarkers, diagnosis is frequently delayed. Understanding FPIES genetics can shed light on disease susceptibility and pathophysiology-key to developing diagnostic, prognostic, preventive and therapeutic strategies. Using a well-characterised cohort of patients we explored the potential genome-wide susceptibility factors underlying FPIES. METHODS: Blood samples from 41 patients with oral food challenge-proven FPIES were collected for a comprehensive whole exome sequencing association study. RESULTS: Notable genetic variants, including rs872786 (RBM8A), rs2241880 (ATG16L1) and rs2289477 (ATG16L1), were identified as significant findings in FPIES. A weighted SKAT model identified six other associated genes including DGKZ and SIRPA. DGKZ induces TGF-ß signalling, crucial for epithelial barrier integrity and IgA production; RBM8A is associated with thrombocytopenia absent radius syndrome, frequently associated with cow's milk allergy; SIRPA is associated with increased neutrophils/monocytes in inflamed tissues as often observed in FPIES; ATG16L1 is associated with inflammatory bowel disease. Coexpression correlation analysis revealed a functional correlation between RBM8A and filaggrin gene (FLG) in stomach and intestine tissue, with filaggrin being a known key pathogenic and risk factor for IgE-mediated food allergy. A transcriptome-wide association study suggested genetic variability in patients impacted gene expression of RBM8A (stomach and pancreas) and ATG16L1 (transverse colon). CONCLUSIONS: This study represents the first case-control exome association study of FPIES patients and marks a crucial step towards unravelling genetic susceptibility factors underpinning the syndrome. Our findings highlight potential factors and pathways contributing to FPIES, including epithelial barrier dysfunction and immune dysregulation. While these results are novel, they are preliminary and need further validation in a second cohort of patients.

New Approaches for Basophil Activation Tests Employing Dendrimeric Antigen-Silica Nanoparticle Composites.

In vitro cell activation through specific IgE bound to high-affinity receptors on the basophil surface is a widely used strategy for the evaluation of IgE-mediated immediate hypersensitivity reactions to betalactams. Cellular activation requires drug conjugation to a protein to form a large enough structure displaying a certain distance between haptens to allow the cross-linking of two IgE antibodies bound to the basophil's surface, triggering their degranulation. However, no information about the size and composition of these conjugates is available. Routine in vitro diagnosis using the basophil activation test uses free amoxicillin, which is assumed to conjugate to a carrier present in blood. To standardize the methodology, we propose the use of well-controlled and defined nanomaterials functionalized with amoxicilloyl. Silica nanoparticles decorated with PAMAM-dendrimer-amoxicilloyl conjugates (NpDeAXO) of different sizes and amoxicilloyl densities (50-300 µmol amoxicilloyl/gram nanoparticle) have been prepared and chemically characterized. Two methods of synthesis were performed to ensure reproducibility and stability. Their functional effect on basophils was measured using an in-house basophil activation test (BAT) that determines CD63+ or CD203chigh activation markers. It was observed that NpDeAXO nanocomposites are not only able to specifically activate basophils but also do so in a more effective way than free amoxicillin, pointing to a translational potential diagnosis.