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Background: The darkling beetle Zophobas morio can be implemented as an alternative in vivo model to study different intestinal colonization aspects. Recently, we showed that its larvae can be colonized by multidrug-resistant Escherichia coli strains administered via contaminated food (for 7 d) for a total experimental duration of 28 d. Method: In the present work, we aimed to shorten the model to 14 d (T14) by administering the previously used CTX-M-15 extended-spectrum β-lactamase-producing ST131 E. coli strain Ec-4901.28 via a single oral administration (5 µL dose of 108 CFU/mL), using a blunt 26s-gauge needle connected to a 250 μL gastight syringe. Force-feeding was performed either without or with (larvae placed on ice for 10 min before injection) anaesthesia. In addition, phage-treated larvae were orally injected with 10 µL of INTESTI bacteriophage cocktail (∼105–6 PFU/mL) on d 4 (T4) and 7 (T7). Results: Growth curve analyses showed that, while larvae rapidly became colonized with Ec-4901.28 (T1, ∼106–7 CFU/mL), only those anaesthetized maintained a high bacterial load (∼102–3 vs. ∼105–6 CFU/mL) and survival rate (76% vs. 99%; P < 0.001) by T14. Moreover, bacteriophage administration to anaesthetized larvae significantly reduced the bacterial count of INTESTI-susceptible Ec-4901.28 at T14 (5.17 × 105 vs. 2.26 × 104, for non-treated and phage-treated larvae, respectively; P = 0.04). Conclusions: The methodological refinements applied to establish the intestinal colonization model simplify the use of Z. morio larvae, facilitate prompt evaluation of novel decolonization approaches and reduce experiments involving vertebrate animals in accordance with the Replacement, Reduction and Refinement principles.

Abstract Background During prolonged FDC therapy, the emergence of FDC non-susceptibility in CRAB has been reported. Here, we report a transmission cluster of FDC-non-susceptible CRAB in four patients, all naïve to FDC treatment, characterized by a premature stop codon and amino acid deletion in the PirA protein. Methods CRAB strains obtained from patients admitted in a single medicine ward of the IRCCS Fondazione Ospedale Maggiore Policlinico between March and July 2024 were analyzed by WGS and antimicrobial susceptibility testing. Phylogenetic analysis was used to assess their genetic relatedness. Results Between March and July 2024, an outbreak of 33 CRAB was observed among hospitalized patients in a single ward at IRCCS. Genomic analysis, available in 29 cases, revealed that 24 isolates belonged to ST208/1806, 4 to ST369, and one to ST195/1816 (according to the Oxford scheme). FDC susceptibility was affected only in the four ST369 isolates (Kirby-Bauer disk diffusion diameter: 13 mm; UMIC® method MIC: 4 mg/L), all characterized by a premature stop codon followed by a 52 amino acid deletion located between the amino acids 377 and 428 of the siderophore-drug receptor PirA. No other relevant mutations were detected in the iron-uptake genes. Core-genome ML tree including ST369 reference strains revealed that the four ST369 isolates were highly related and formed a distinct cluster (SNP distance: 3 [IQR: 1–6]). Of note, the four isolates were collected from four FDC-naïve individuals, two experiencing a CRAB-mediated infection. Conclusions Our findings alert about the circulation of clones carrying modified siderophore-drug receptors without evidence of previous FDC treatment and support the importance of testing FDC susceptibility appropriately before its administration.

Abstract Background Microglia play crucial roles in maintaining neuronal homeostasis but have been implicated in contributing to amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). However, the role of microglia in ALS/FTD remains incompletely understood. Methods Here, we generated highly enriched cultures of VCP mutant microglia derived from human induced pluripotent stem cells (hiPSCs) to investigate their cell autonomous and non-cell autonomous roles in ALS pathogenesis. We used RNA-sequencing, proteomics and functional assays to study hiPSC derived VCP mutant microglia and their effects on hiPSC derived motor neurons and astrocytes. Results Transcriptomic, proteomic and functional analyses revealed immune and lysosomal dysfunction in VCP mutant microglia. Stimulating healthy microglia with the inflammatory inducer lipopolysaccharide (LPS) showed partial overlap with VCP mutant microglia in their reactive transformation. LPS-stimulated VCP mutant microglia displayed differential activation of inflammatory pathways compared with LPS-stimulated healthy microglia. Conserved gene expression changes were identified between VCP mutant microglia, SOD1 mutant mice microglia, and postmortem ALS spinal cord microglial signatures, including increased expression of the transmembrane glycoprotein GPNMB. While knockdown of GPNMB affected inflammatory and phagocytosis processes in microglia, this was not sufficient to ameliorate cell autonomous phenotypes in VCP mutant microglia. Secreted factors from VCP mutant microglia were sufficient to activate the JAK-STAT pathway in hiPSC derived motor neurons and astrocytes. Conclusions VCP mutant microglia undergo cell autonomous reactive transformation involving immune and lysosomal dysfunction that partially recapitulate key phenotypes of microglia from other ALS models and post mortem tissue. These phenotypes occur independently of GPNMB. Additionally, VCP mutant microglia elicit non cell autonomous responses in motor neurons and astrocytes involving the JAK-STAT pathway.

The Taraba Sound is situated north of the Strait of Magellan within the Chilean fjord and channel system. In this inlet, samples were collected at three beach levels (supralittoral, mesolittoral and infralittoral) along four transects perpendicular to the coastline. A total of 2,754 specimens were collected and identified, representing seven phyla and 24 operational taxonomic units (OTUs). The dominant components across the four beaches were bivalves and crustaceans, with Choromytilus chorus and Paramoera cf. fissicauda contributing significantly to zone similarity. Beaches 2 and 4, located adjacent to river mouths, had lower salinity and consequently lower species richness, abundance, diversity and evenness, particularly in the supra- and mesolittoral zones where freshwater influence is more pronounced. Our study represents the first comprehensive characterisation of intertidal benthic invertebrate biodiversity in the region. Its findings provide valuable baseline data for future research, especially considering the anticipated increase in aquaculture concession applications and the establishment of offshore sea farming centres in the region.

El artículo analiza las políticas culturales implementadas entre 2007 y 2015 en relación con el Instituto Saturnino Unzué de Mar del Plata, Provincia de Buenos Aires, Argentina, destacando la intervención estatal y la colaboración interjurisdiccional en su preservación y refuncionalización. Se estudian las acciones de diversos actores, resaltando la necesidad de políticas más integrales y sostenibles para garantizar la conservación de este patrimonio cultural. El trabajo subraya la importancia de las intervenciones conjuntas para mantener vivo este espacio emblemático, que sigue siendo un centro cultural activo y significativo para la comunidad local, provincial y nacional.

Abstract Background Despite advances in screening and therapy, breast cancer (BC) remains the predominant cancer in women globally. Dysregulation of microRNAs (miRNAs) is pivotal in carcinogenesis across various cancers, including BC. Evidence indicates that miR‐1307‐3p is upregulated in BC tumors, yet its target genes are not fully elucidated. This study aimed to explore how miR‐1307‐3p regulates BC proliferation, migration, invasion, and angiogenesis and to identify potential target genes. Methods Basal miR‐1307‐3p levels were quantified in BC cell lines MDA‐MB‐231 and MCF‐7, as well as MCF‐10A using quantitative real‐time reverse transcription‐PCR (RT‐qPCR). The impact of miR‐1307‐3p inhibition on BC cell proliferation, migration, invasion, and angiogenesis was assessed. Nine miRNA‐target prediction databases identified potential miR‐1307‐3p targets. Target expression was validated using RT‐qPCR, Western blot, and dual‐luciferase reporter assays. MiR‐1307‐3p was overexpressed in MDA‐MB‐231 and MCF‐7 compared to MCF‐10A. Results Inhibiting miR‐1307‐3p significantly reduced BC cell proliferation, migration, invasion, and angiogenesis. Bioinformatics analysis identified 17 potential miR‐1307‐3p targets, with protamine 2 (PRM2) overexpression confirmed via Western blot and dual‐luciferase assays. Conclusion MiR‐1307‐3p overexpression in BC promotes proliferation, migration, invasion, and angiogenesis. PRM2 emerges as a novel miR‐1307‐3p target in BC.

Abstract The historical restriction of magnetic resonance imaging (MRI) for patients with cardiac implantable electronic devices (CIEDs) has been lifted by certified MRI-conditional systems in recent years. Mixed-brand CIED systems consisting of a generator from one manufacturer and at least one lead from another manufacturer are not certified for MRI. We evaluated the temporal trend in the prevalence of mixed-brand systems in the era of MRI-conditional systems. Data were analyzed on 5853 CIEDs implanted de novo between 2012 and 2022 in 81 Italian centers linked to the nationwide Home Monitoring Expert Alliance network. The percentage of mixed-brand implants was calculated by device type (pacemaker, implantable cardioverter-defibrillator [ICD], cardiac resynchronization therapy [CRT] device) and over time. A mixed-brand system was implanted in 4.1% (95% CI, 3.6-4.6%) of analyzed patients or, by device type, in 4.5% (3.5-5.7%) of pacemaker patients, 1.1% (0.7-1.7%) of ICD patients, and 6.8% (5.7-7.9%) of CRT pacemaker/defibrillator patients (p

Abstract Objective We investigated the effects of adding regions to current dissemination in space (DIS) criteria for multiple sclerosis (MS). Methods Participants underwent brain, optic nerve, and spinal cord MRI. Baseline DIS was assessed by 2017 McDonald criteria and versions including optic nerve, temporal lobe, or corpus callosum as a fifth region (requiring 2/5), a version with all regions (requiring 3/7) and optic nerve variations requiring 3/5 and 4/5 regions. Performance was evaluated against MS diagnosis (2017 McDonald criteria) during follow‐up. Results Eighty‐four participants were recruited (53F, 32.8 ± 7.1 years). 2017 McDonald DIS criteria were 87% sensitive (95% CI: 76–94), 73% specific (50–89), and 83% accurate (74–91) in identifying MS. Modified criteria with optic nerve improved sensitivity to 98% (91–100), with specificity 33% (13–59) and accuracy 84% (74–91). Criteria including temporal lobe showed sensitivity 94% (84–98), specificity 50% (28–72), and accuracy 82% (72–90); criteria including corpus callosum showed sensitivity 90% (80–96), specificity 68% (45–86), and accuracy 85% (75–91). Criteria adding all three regions (3/7 required) had sensitivity 95% (87–99), specificity 55% (32–76), and accuracy 85% (75–91). When requiring 3/5 regions (optic nerve as the fifth), sensitivity was 82% (70–91), specificity 77% (55–92), and accuracy 81% (71–89); with 4/5 regions, sensitivity was 56% (43–69), specificity 95% (77–100), and accuracy 67% (56–77). Interpretation Optic nerve inclusion increased sensitivity while lowering specificity. Increasing required regions in optic nerve criteria increased specificity and decreased sensitivity. Results suggest considering the optic nerve for DIS. An option of 3/5 or 4/5 regions preserved specificity, and criteria adding all three regions had highest accuracy.

Abstract Juvenile‐onset Huntington's disease (HD) is a rare subset of HD with symptom‐onset before the age of 18. In contrast to the adult population, children present early‐on with behavioral, psychiatric, and cognitive symptoms, in addition to a diverse spectrum of movement disorders. This poses a distinct challenge in diagnosis and management. We here describe the spectrum of movement disorders, accompanied with detailed video recordings, in seven cases of juvenile‐onset HD. Our findings highlight early cognitive and behavioral symptoms, preceding motor symptoms. The diverse movement disorder phenotypes included dystonia, Parkinsonism, myoclonus, and chorea, findings which underscore the heterogeneity of presenting symptoms.

The aim of this study was to identify pitfalls in ovarian tissue cryopreservation protocol from referral to surgical procedures and to analyze factors associated with chemotherapy exposure of the cryopreserved tissue and decreased ovarian function in a cohort of young girls at high risk of infertility. The study population comprised 200 girls eligible for ovarian tissue cryopreservation between 2002 and 2020 at the Children's Hospital of the University Central Hospital of Helsinki (Finland). Analyses included the evaluation of the proportion of patients who underwent ovarian tissue cryopreservation, factors associated with patient selection and timing of ovarian tissue cryopreservation, and ovarian function during long-term follow-up in relation to oncological treatments. Lack of counseling was identified as the major reason for not receiving ovarian tissue cryopreservation. A longer interval from scheduling gonadotoxic therapy to cryopreservation correlated with a higher exposure to alkylating agents of the ovarian tissue. The long-term ovarian function was mainly influenced by age at the time of gonadotoxic treatment. Current selection criteria for ovarian tissue cryopreservation should be implemented in order to stratify patients at risk of infertility and timely identify those at higher risk, especially in relation to age and pubertal stage. Efforts to increase healthcare providers’ awareness and facilitate guided timing in relation to the treatment protocols are needed to guarantee early access to ovarian tissue cryopreservation for all patients at high risk of infertility. Lay summary Girls affected by cancer may undergo aggressive treatments to cure the disease, such as chemotherapy and radiotherapy, which can harm the ovaries. These treatments often result in the destruction of the ovaries and infertility. Nowadays, an option exists to help these girls recover their fertility after the cancer has been successfully treated. A fragment or an entire ovary can be removed by surgery and frozen before the treatments begin. When the girls are well and wish as adults to start a family, this fragment can be reimplanted back into the remaining ovary to restore fertility. This technique is called ovarian tissue cryopreservation. In this article, we report on the experience of ovarian tissue cryopreservation at the Children’s Hospital in Finland for 200 girls undergoing cancer treatment with a high risk of subsequent infertility. We analyzed the proportion of girls who had ovarian tissue cryopreservation performed among those exposed to aggressive treatments and investigated the reasons why some of them did not undergo the procedure. We also examined the time passing before the procedure was done to identify potential avoidable sources of delay. Finally, we explored how the ovaries of all the girls functioned during the time after the cancer diagnosis and looked at this in relation to their cancer treatments and ovarian tissue freezing.

Objectives. This study aimed to compare the audiological benefits of a non-implantable wearable option for a bone conduction (BC) implant mounted on an arch (SoundArc) to those of traditional BC hearing aids (HAs) mounted on eyeglasses (BCHAs) in patients with moderate to severe conductive or mixed hearing loss. Methods: A preliminary cross-sectional observational prospective cohort study was conducted in the Tertiary Audiological Department, University Hospital. Fourteen adults with conductive or mixed hearing loss (PTA at 0.5-1-2-4 KHz = 67 ± 15 dB HL) who had been wearing conventional BCHAs mounted on eyeglasses for at least 3 years and had declined surgical implantation of a bone conduction hearing implant (BCHI) were included in the study. Unaided and aided pure-tone air conduction (AC) and bone conduction (BC) thresholds, as well as speech tests in quiet and noise, were recorded at baseline and in two different settings: with a BCHI mounted on SoundArc® and with their own BCHAs mounted on eyeglasses using two couplers. Participants completed questionnaires in both conditions, including the International Inventory for Hearing Aids (IOI-HA), the Hearing Handicap Inventory for Adults/Elderly (HHIA/E), the Speech, Spatial, and Qualities of Hearing Scale (SSQ), a 10-point visual analog scale (VAS), and the Fatigue Impact Scale (FIS). Results: A significant functional gain was observed in both settings (p = 0.0001). Better speech perception in quiet and noise was observed with SoundArc compared to conventional BCHAs on eyeglasses (improvements in word repetition scores in noise: +19.3 at SNR +10 dB, p = 0.002; +12.1 at SNR 0 dB, p = 0.006; and +11.4 at SNR −10 dB, p = 0.002). No significant differences were found in IOI-HA, FIS, and HHIA/E scores. However, significantly better SSQ scores were reported for SoundArc in all domains (p = 0.0038). Conclusions: Although patients were accustomed to using BCHAs mounted on eyeglasses, the bone conduction wearable option of the BCHI (SoundArc) proved to be a viable alternative for adult patients with conductive or mixed hearing loss who are unable or unwilling to undergo BCHI surgery.

Abstract Background Pediatric residency in Italy has undergone significant changes in recent years, with a notable increase in the number of pediatric residents. Exploring the implications of this increase, highlighting disparities in training opportunities, and discussing the broader socio-economic impacts on pediatric healthcare, represent a crucial consideration for the healthcare system in the future. Main Body The Italian National Association of Pediatric Residents (“Osservatorio Nazionale Specializzandi in Pediatria”, ONSP) conducted an extensive survey among pediatric residents to assess the current state of pediatric residency. Key findings indicate that 50% of respondents believe the number of residents is excessive for the available training opportunities, leading to concerns about the quality of education and hands-on experience. Despite the increased number of residents, the workload has increased, but up to one-third of residents feel that the autonomy provided by their programs is insufficient. Significant disparities in training quality were found across different regions, with notable shortages in neonatology and pediatric emergency departments. Research opportunities are also limited, with only 17% of residents finding the time allocated to research satisfactory, especially in central and southern Italy. Conclusion The increase in pediatric residents presents both challenges and opportunities. Addressing these challenges through strategic reforms, such as implementing standardized national curricula, investing in training resources and mobility programs, and enhancing research opportunities, is crucial for the future of pediatric residency in Italy. Ensuring high-quality training for all residents is an ethical and practical obligation that will significantly impact pediatric healthcare.

Abstract Coronavirus disease 2019 (COVID-19) has been associated with brain functional, structural, and cognitive changes that persist months after infection. Most studies of the neurologic outcomes related to COVID-19 focus on severe infection and aging populations. Here, we investigated the neural activities underlying COVID-19 related outcomes in a case-control study of mildly infected youth enrolled in a longitudinal study in Lombardy, Italy, a global hotspot of COVID-19. All participants (13 cases, 27 controls, mean age 24 years) completed resting-state functional (fMRI), structural MRI, cognitive assessments (CANTAB spatial working memory) at baseline (pre-COVID) and follow-up (post-COVID). Using graph theory eigenvector centrality (EC) and data-driven statistical methods, we examined differences in ECdelta (i.e., the difference in EC values pre- and post-COVID-19) and Volumetricdelta (i.e., the difference in cortical volume of cortical and subcortical areas pre- and post-COVID) between COVID-19 cases and controls. We found that ECdelta significantly between COVID-19 and healthy participants in five brain regions; right intracalcarine cortex, right lingual gyrus, left hippocampus, left amygdala, left frontal orbital cortex. The left hippocampus showed a significant decrease in Volumetricdelta between groups (p = 0.041). The reduced ECdelta in the left amygdala associated with COVID-19 status mediated the association between COVID-19 and disrupted spatial working memory. Our results show persistent structural, functional and cognitive brain changes in key brain areas associated with olfaction and cognition. These results may guide treatment efforts to assess the longevity, reversibility and impact of the observed brain and cognitive changes following COVID-19.

Conflict of interest: Yes, This work was supported by Brazilian Society of Hepatology and Instituto Brasileiro do Fígado - IBRAFIG. Introduction and Objectives: Previous studies have identified risk factors associated with recurrent primary sclerosing cholangitis (rPSC) after liver transplantation (LT) in Caucasians. There is paucity of data regarding rPSC in multiethnic Latin patients. Objectives: To investigate rPSC frequency and its associated risk factors in a highly admixed population from Brazil. Patients / Materials and Methods: The Brazilian Cholestasis Study Group database was retrospectively reviewed for including primary sclerosing cholangitis (PSC) patients who underwent LT. Primary outcome was rPSC. Results and Discussion: A total of 96 patients were included, 60% males, mean age 32 ± 13 years. After a follow-up of 90 months (interquartile range 39-154), rPSC occurred in 29 (30%) of the participants. There were no statistically significant associations between rPSC and age, gender, concurrent or de novo inflammatory bowel disease, MELD score at the time of LT or allograft rejection. The only factor associated with an increased risk of disease recurrence was time after LT. Conclusions: In Brazilian PSC patients who underwent LT, one-third had rPSC. Longer time after LT was associated with rPSC diagnosis.

Conflict of interest: No Introduction and Objectives: Primary biliary cholangitis is a chronic and progressive autoimmune liver disease, whose prognosis can be improved by normalizing alkaline phosphatase and bilirubin. While ursodeoxycholic acid (UDCA) is first line standard of care, approximately 40% of patients exhibit incomplete response. We aimed to identify prognostic markers for deep response to UDCA therapy at presentation. Patients / Materials and Methods: Data from the Brazilian Cholestasis Study Group cohort were analyzed retrospectively. Patients were assessed for deep response (defined as normalization of alkaline phosphatase and bilirubin) after 1 year of UDCA treatment. With the purpose of selecting the set of relevant variables related to the deep response for a parsimonious multivariate model, we applied the Varrank algorithm. Additionally, the performance of the UDCA response score in predicting deep response was evaluated. Results and Discussion: A total of 297 patients were analyzed, with 57.2% achieving an adequate response according to the Toronto criteria, while 22.9% reached deep response. Cirrhosis (OR 0.460; 95% CI 0.225-0.942; p=0.034) and elevated baseline alkaline phosphatase levels (OR 0.629; 95% CI 0.513-0.770; p

Conflict of interest: Yes, FONDECYT #1241450 Introduction and Objectives: Although Latinos living in the United States are at higher risk of steatotic liver disease (SLD), information from Latin American countries is extremely scarce. We aimed to characterize SLD in Latin America and explore the role of the common genetic variants in this region. Patients / Materials and Methods: Cross-sectional multicenter study including individuals with SLD who undergo liver biopsy or transient elastography (TE) between 2003–2024. TE thresholds were established as follows: significant fibrosis (F2) ≥8.2 kPa, advanced fibrosis (F3) ≥9.7 kPa, and cirrhosis (F4) ≥13.6 kPa. Analyses included logistic binary regression. Results and Discussion: We included 2,159 patients (93.7% metabolic dysfunction-associated SLD and 6.3% alcohol-associated liver disease) from 13 centers in 5 countries (Argentina, Brazil, Chile, Mexico, and Peru). Mean age was 54.3±14.3 years old, 58.8% were female, and 60.2% had a liver biopsy. Around 18% had significant fibrosis, 18.7% advanced fibrosis, and 16.2% had cirrhosis. Additionally, 21.8% were homozygous carriers of the rs738409 risk polymorphism (PNPLA3 I148M variant), 42.5% were heterozygotes, and 35.7% were non-carriers. In an adjusted multivariable model, only age (odds ratio [OR]:1.03; 95%CI:1.01–1.04; p=0.030), body mass index (BMI) (OR:1.04; 95%CI:1.01–1.08, p=0.008), prediabetes/diabetes (OR:2.41; 95%CI:1.48–3.91, p

Background: Falls amongst the elderly represent a global public health challenge because of their potential to cause illness, death, and reduce the autonomy of this group. They also impact the emotional, family, social and economic well-being of those involved. Various strategies to prevent falls have been reported in the literature, focusing mainly on addressing individual risk factors, and on the continuous assessment of the risk of falls in older people. Objective: This study evaluated user satisfaction and acceptability of a comprehensive model, implemented in the community, to prevent falls amongst independent older adults aged 65 years and above. It sought to capture both the perceptions of the individuals who received the intervention and of the interventionists who implemented it. The study protocol was registered at ClinicalTrials.gov in November 2020 (ID: NCT04313062). Design: Qualitative, exploratory study using a case study design. The evaluation of the intervention followed the recommendations proposed by the Medical Research Council for complex interventions. Methods and participants: In the period between April 2021 to April 2022, 11 semi-structured interviews were conducted with independent older adults between 65 and 80 years of age who participated in the implementation of the comprehensive model in Santiago, Chile. Data were also collected with eight interventionists through: three semi-structured interviews at the beginning of the intervention; and two focus groups with seven interventionists at the end of the implementation of the model. The team members undertook a content analysis of the data collected. Results: Three themes emerged to account for the satisfaction and acceptability of the intervention with the model on the part of the participants and interventionists: (1) Previous experience of older persons and interventionists; (2) The older person-interventionist encounter and its context; and (3) Identification of facilitators, strengths and challenges for the implementation of the model. The results show a positive assessment of the model, highlighting the value of the social contact derived from the intervention by both participants and interventionists. Although the model involved an individual intervention, the participants’ accounts indicate that it reached out to others, including family members and other elderly acquaintances. Moreover, the interventionists helped identify challenges in implementation and made recommendations to strengthen the model. Conclusion: The evaluation of satisfaction and feasibility of implementing the model showed positive results that will nurture the next phase of development of this model, which involves scaling up the intervention.

Abstract Background Elephant endotheliotropic herpesvirus (EEHV) infection is the most common cause for lethal hemorrhagic disease in captive juvenile Asian elephants (Elephas maximus). Although EEHV1 is known as the most likely cause of fatal haemorrhagic disease in Asian elephants, EEHV5 was lately involved in lethal cases of haemorrhagic disease in captive elephants. Case presentation Here we report the first death of a four-year old Asian elephant diagnosed with EEHV5 in Germany. Molecular diagnosis yielded detection of EEHV5 DNA in all tested tissues. Histopathological examination revealed typical features of hemorrhagic disease in all examined organs. EEHV5 was sequenced from total DNA isolated from heart tissue by Illumina and Nanopore sequencing. Sequencing data showed 3,881 variants, distributed across the entire genome, compared to the published EEHV5 sequence. Conclusions We have detected EEHV5 in a fatal disease case of a male Asian elephant. Whole genome sequencing revealed substantial differences of our DNA isolate compared to available EEHV5 sequences. This report of fatal haemorrhagic disease associated with EEHV5 infection should raise awareness for EEHV5 as an important elephant pathogen. Genome sequencing and downstream SNPs analysis will further encourage future research to understand genetic diversity, pathogenesis and virulence of EEHVs with respect to developing new diagnostic methods, prophylactic strategies, and implementation of surveillance and control measures.

Abstract Background HIV infection can cause malabsorption and rapid utilization of nutrients. A randomized trial of multivitamin supplementation among people living with HIV/AIDS (PLWHA) initiating antiretroviral therapy (ART) in Tanzania was stopped early due to increased alanine aminotransferase (ALT) concentrations in the multiple recommended dietary allowances (RDA) multivitamin group. We conducted detailed analysis to assess the effect of multivitamins on ALT elevations and evaluate whether subgroups of PLWHA have greater hepatotoxicity risks associated with the use of high-dose multivitamins. Methods We utilized data from a randomized, double-blind trial conducted in 2006–2009 that assessed the effect of high-dose multivitamins that contained vitamin B complex, vitamin C, and vitamin E at multiple RDA as compared to standard-dose multivitamins containing single RDAs among adults initiating ART in Tanzania. We evaluated the effect of high-dose multivitamins on incident mild/moderate ALT elevations > 40 IU/L, persistent ALT elevations > 40 IU/L (2 + clinic visits), and severe ALT elevations > 200IU/L using Cox proportional hazard models. We then evaluated effect modification by patient characteristics to determine if subgroups of PLWHA experienced different magnitudes of risk for ALT elevations associated with high-dose multivitamins. Results High-dose multivitamins increased the risk of incident mild/moderate ALT elevations > 40 IU/mL as compared to standard-dose multivitamins (hazard ratio (HR): 1.41; 95%CI: 1.26,1.58) as well as incident sustained mild/moderate ALT elevations (HR: 1.19; 95%CI: 1.04,1.36), but there was no overall effect on severe ALT elevations (HR: 1.44; 95% CI: 0.91,2.28). There was no evidence that the effect of high-dose multivitamins on any or sustained mild/moderate ALT elevations was modified by any patient characteristic. However, CD4 T-cell count was found to modify the effect of high-dose multivitamins on severe ALT elevations (p-value for interaction:0.01). Among participants with a baseline CD4 T-cell count ≤ 100 cells/µL, individuals receiving high-dose multivitamins had 3.74 times (95%CI: 1.52–9.17) the risk of incident severe ALT elevations compared to standard-dose multivitamins, while participants with CD4 T-cell counts > 100 cells/µL, appeared to have no effect of high-dose multivitamins on severe ALT elevations (HR:0.92; 95% CI: 0.50,1.67). Conclusions High-dose RDA multivitamin supplementation increased the incidence of any mild to moderate ALT elevations among adults starting ART in Tanzania and the magnitude of the risk does not appear to differ by patient characteristics. However, immunocompromised PLWHA with CD4 T-cell counts

Abstract Mesenchymal stem/stromal cells (MSCs) have emerged as a promising tool in the field of regenerative medicine due to their unique therapeutic properties as they can differentiate into multiple cell types and exert paracrine effects. However, despite encouraging results obtained in preclinical studies, clinical trials have not achieved the same levels of efficacy. To improve the therapeutic properties of MSCs, several strategies have been explored. Therefore, in this review, the therapeutic properties of MSCs will be analyzed, and an update and overview of the most prominent approaches used to enhance their therapeutic capabilities will be provided. These approaches include using drugs, molecules, strategies based on biomaterials, and modification parameters in culture. The strategy described shows several common factors among those affected by these strategies that lead to an enhancement of the MSCs therapeutic properties such as the activation of the PI3K/AKT pathway and the increased expression of Heat Shock Proteins and Hypoxia-Inducible Factor. The combined effect of these elements shift MSCs towards a glycolytic state, suggesting this shift is essential for their enhancement. Graphical abstract

Abstract Background Women with high-risk breast lesions, such as atypical hyperplasia (AH) or lobular carcinoma in situ (LCIS), have a 4- to tenfold increased risk of breast cancer compared to women with non-proliferative breast disease. Despite high-quality data supporting chemoprevention, uptake remains low. Interventions are needed to break down barriers. Methods The parent trial, MiCHOICE, is a cluster randomized controlled trial evaluating the effectiveness and implementation of patient and provider decision support tools to improve informed choice about chemoprevention among women with AH or LCIS. For this pre-implementation analysis, 25 providers participated in semi-structured interviews prior to accessing decision support tools. Interviews sought to understand attitudes/beliefs and barriers/facilitators to chemoprevention. Results Interviews with 25 providers (18 physicians and 7 advanced practice providers) were included. Providers were predominantly female (84%), white (72%), and non-Hispanic (88%). Nearly all providers (96%) had prescribed chemoprevention for eligible patients. Three themes emerged in qualitative analysis. The first theme describes providers’ confidence in chemoprevention and the utility of decision support tools. The second theme elucidates barriers to chemoprevention, including time constraints, risk communication and perceptions of patients’ fear of side effects and anxiety. The third theme is the need for early implementation of decision support tools. Conclusions This qualitative study suggests that providers were interested in the early inclusion of decision aids (DA) in their chemoprevention discussion workflow. The DAs may help overcome certain barriers which were elucidated in these interviews, including patient level concerns about side effects, clinic time constraints and difficulty communicating risk. A multi-faceted intervention with a DA as one active component may be needed. Trial registration This trial was registered with the NIH clinical trial registry, clinicaltrials.gov, NCT04496739.

Objectives: Tilia viridis (Bayer) Simonk. (Malvaceae) is widely distributed in Argentina and employed for its tranquilizing properties. Other species of the genus (Tilia europaea L., Tilia cordata Mill., Tilia platyphyllos Scop.) have been traditionally used for the treatment of epilepsy. Epilepsy affects approximately 65 million people worldwide and is characterized by an imbalance between excitatory and inhibitory processes in the brain, leading to unpredictable, unprovoked, recurrent seizures. Current pharmacological interventions often present mild to moderately severe side effects. Epilepsy has been associated with oxidative and nitrative stress as well as neuroinflammation. Herbal medicine therapies may offer new treatment options with multi-target antioxidant and anticonvulsant effects for patients whose seizures remain uncontrolled, potentially providing cost-effective solutions for individuals worldwide suffering from uncontrolled epilepsy.The aim of this study was to demonstrate the anticonvulsant activity of a standardized T. viridis aqueous extract (TE).Methods: Study of the constituents of TE, TE’s antioxidant and anticonvulsant activities and toxicity, and analysis of the possible relation between the potential activities and the compounds present in the extract. In order to demonstrate TE’s anticonvulsant activity a zebrafish model was used. The study also assessed TE’s toxicity and antioxidant activity. To standardize the extract, total polyphenols and flavonoids were quantified and specific flavonoids were identified and quantified using HPLC-MS/MS and HPLC-UV.Results: TE exhibited anticonvulsant activity at low concentrations and demonstrated antioxidant effects by scavenging free radicals, exhibiting superoxide dismutase and peroxidase- like activities, as well as inhibiting lipoperoxidation. These actions can be attributed to the presence of polyphenols, particularly flavonoids.Conclusion: TE holds promise as a complementary herbal medicine in the treatment of epilepsy and may also offer benefits for other neuropathies associated with oxidative stress, such as Parkinson’s disease and Alzheimer’s disease.

The description of diagnostic morphological characters and DNA barcoding of fish larvae from nine species of the carangid family are provided from specimens collected during a weekly zooplankton time-series (2016–2017) at Cabo Pulmo National Park, Gulf of California, Mexico. Five nominal species (Caranx sexfasciatus, C. caballus, Naucrates ductor, Selar crumenophthalmus, and Selene peruviana) and three morphotypes of Decapterus spp. and one of Caranx spp. were identified and separated based on morphological, meristic, and pigmentary diagnostic characters. All larvae were genetically sequenced for a fragment of the cytochrome c oxidase subunit I mitochondrial gene. Sequences of larval Caranx and Decapterus showed high genetic similarity (> 99%), low intraspecific divergence (< 1%), and an interspecific divergence between 6% and 11%, allowing the discrimination of diagnostic pigmentation patterns of fish larvae among three sibling species from each genus: Caranx (C. caballus, C. caninus, and C. sexfasciatus) and Decapterus (D. macarellus, D. macrosoma, and D. muroadsi). DNA barcoding supported the presence of Caranx caballus, C. caninus, C. sexfasciatus, Decapterus macarellus, D. muroadsi, Selar crumenophthalmus, and Selene peruviana, and for the first time Naucrates ductor and D. macrosoma at the CPNP. Abundance of these nine species (confirmed molecularly) was estimated throughout the 2016–2017 weekly time series. Decapterus macarellus and Caranx caninus were the most abundant species. The morphological and molecular taxonomic methods allowed us to infer the species number and abundance of these commercial species at the CPNP to improve conservation in protected areas and fishery management.

El objetivo de este artículo es analizar las percepciones de las mujeres agricultoras en México para identificar su interpretación en relación con: (1) las principales actividades que realizan en el campo, (2) el uso de maquinaria y las labores agrícolas, (3) la valoración de su trabajo, (4) las oportunidades laborales, (5) los recursos y apoyos necesarios para mejorar la producción, (6) la toma de decisiones en la siembra, (7) el acceso a la capacitación y desarrollo, (8) las implicaciones del cambio climático, (9) los desafíos que enfrentan en el campo y (10) las propuestas para fomentar su participación en el sector agrícola. Es un estudio cualitativo; la obtención de información se realizó mediante entrevistas semiestructuradas a once mujeres originarias de municipios de Hidalgo, Puebla, Estado de México y de la alcaldía Xochimilco en la Ciudad de México. Se encontró que las mujeres expresaron que su trabajo no es valorado, se mantiene la creencia de que ellas son proveedoras de cuidados y son encargadas de las labores domésticas; muchas productoras a pequeña escala no cuentan con ingresos económicos y las que trabajan para algún patrón se les paga por día, y reconocen que ganan menos que los hombres. Además, las tierras en su mayoría son propiedad de los esposos o las rentan a varones; las mujeres expresan que no tienen derecho a heredar la tierra y que su toma de decisiones todavía es un asunto reservado, aunado a que se considera que el campo es masculino. Se concluye que es necesario apoyar al campo mexicano con perspectiva de género pensado que las mujeres son la fuerza más poderosa de la soberanía alimentaria.

Abstract Background Aggressive malignancies, such as pancreatic cancer, are increasingly impacting young, female populations. Our investigation centered on whether the observed trends in cancer incidence were unique to pancreatic cancer or indicative of a broader trend across various cancer types. To delve deeper into this phenomenon, we analyzed cancer incidence trends across different age and sex groups. Furthermore, we explored differences in cancer incidence within specific young subgroups aged 18 to 26 and 27 to 34, to better understand the emerging incidence trend among young individuals. Methods This study collected cancer incidence data from one of the Surveillance, Epidemiology, and End Results cancer registry databases (SEER22), with 10,183,928 total cases from 2000 to 2020. Data were analyzed through Joinpoint trend analysis approach to evaluate sex- and age-specific trends in cancer incidence. Exposure rates were reported as Average Annual Percentage Changes (AAPCs). Results The analysis revealed significant age and sex-specific disparities, particularly among individuals aged 18–26 and 27–34. Pancreatic cancer incidence rates increased more in females aged 18–26 (AAPC, 9.37% [95% CI, 7.36–11.41%]; p

Introdução: Os casos de síndrome gripal (SG) e síndrome respiratória aguda grave (SRAG) vem aumentando, sabe-se que pacientes oncológicos estão entre os grupos de risco. Um hospital oncológico de grande porte, estabeleceu um fluxo desde o início para o diagnóstico etiológico dos pacientes com SG e SRAG com pesquisa de antígeno COVID-19 (Kovalent do Brasil Ltda) em swab de nasofaringe (NF) e para os pacientes internados com pesquisa de antígeno COVID-19 negativa, um novo swab de NF é coletado e encaminhado para o IAL/ SP para realizar painel viral (Protocolo CDC de Vírus Respiratório/Atlanta/EUA). Objetivo: Descrever os agentes etiológicos encontrados nos pacientes com diagnósticos de SG e SRAG que procuraram o serviço no período de janeiro/23 a abril/24. Método: Estudo descritivo da frequência dos vírus respiratórios nos pacientes atendidos na unidade pronto atendimento (PA) e internação. O hospital conta com 162 leitos de sendo 10 de UTI, com média mensal de 1.100 internações e 2.500 atendimentos na unidade de PA. Cerca de 75% dos atendimentos são direcionados a pacientes oncológicos e o restante ao tratamento de patologias ginecológicas benignas, reprodução humana e vítimas de violência sexual. Resultados: No período de jan/23 a abr/24, 165 pacientes tiveram diagnóstico de SG ou SRAG, desses 89 (53,9%) tiveram o diagnóstico de SG, sem indicação de internação hospitalar, dos quais 29 (32%) apresentaram pesquisa de antígeno COVID-19 positivo. Os demais 76 (46,1%) pacientes foram internados, dos quais 16 (21%) apresentaram pesquisa de antígeno COVID-19 positivo. Dentre os 60 (79%) das pacientes com pesquisa de antígeno COVID-19 negativa, em 58 (76,3%) foi realizado painel viral, sendo 48 (63,1%) com resultado negativo e 10 (13,1%) com resultado positivo (3 casos positivo para rinovírus, 2 casos positivos para COVID-19, 1 caso positivo para VSR, 1 caso positivo para FLU A, 1 caso positivo para FLU B, 1 caso positivo para COVID-19 + adenovírus e 1 caso positivo para VRS + rinovírus.). A incidência de SG e SRAG no períodos foi de 412,5 casos por 100.000 atendimentos no PA. Conclusão: Dentre os casos de SG e SRAG, COVID-19 é o agente mais prevalente (27%). Os meses mar/23, abr/23, fev/24 e mar/24 foram os meses de maior incidência de casos de SG e SRAG, como também do vírus COVID-19. Dentre os isolados dos outros tipos de vírus, não foi possível determinar uma sazonalidade. No período, não foi possível diagnosticar caso de transmissão hospitalar.

Las transformaciones en las ciudades producidas desde el inicio de la revolución industrial son una de las principales amenazas para la salvaguarda del patrimonio urbano. Estas circunstancias han demandado la elaboración de estrategias y nuevos enfoques orientados a lograr un desarrollo sustentable del patrimonio cultural. Al respecto, en las últimas décadas el vínculo entre el patrimonio cultural y las diferentes dimensiones de la sustentabilidad ocupa uno de los puntos centrales de la agenda patrimonial de organizaciones nacionales e internacionales, así como de investigadores de todo el mundo. De las diferentes dimensiones de la sustentabilidad identificadas, es la sociocultural la que mayor incidencia tiene en la salvaguardia del patrimonio cultural. En el abordaje del patrimonio cultural desde la perspectiva de la sustentabilidad los procesos de evaluación mediante el uso de indicadores presentan amplias ventajas para la formulación, análisis y seguimiento de las políticas culturales y los procesos de toma de decisiones en la gestión. En este contexto, existe un amplio espectro de propuestas de indicadores de sustentabilidad que son aplicados, desde diferentes enfoques, a una diversidad de objetos y de contextos. No obstante, en general se observa que, en la mayoría de las propuestas, se formulan indicadores cuantitativos con una fuerte incidencia de las dimensiones económica y ambiental, presentando limitaciones para abordar aspectos más sensibles del patrimonio cultural, como las valoraciones y percepciones, que por tener un carácter subjetivo no son fácilmente cuantificables. Por ello, en el presente trabajo se presenta una propuesta metodológica para la evaluación de la sustentabilidad sociocultural del patrimonio urbano en ciudades medias mediante un conjunto de indicadores formulados desde un enfoque cualitativo, diseñada y testeada en una ciudad intermedia de Argentina (Olavarría). La metodología propuesta permite medir los avances y retrocesos del patrimonio en pos de su sustentabilidad en el mediano plazo. Además, facilita la integración de evaluaciones de distintas disciplinas e incorpora la mirada de los principales actores de la patrimonialización y la gestión del patrimonio, así como posibilita la realización de un diagnóstico de la situación del patrimonio urbano en las áreas de estudio. The transformations in cities since the beginning of the industrial revolution are one of the main threats to safeguarding urban heritage. These circumstances have demanded the elaboration of strategies and new approaches to achieve the sustainable development of cultural heritage. In this respect, in recent decades, the link between cultural heritage and the different dimensions of sustainability has been at the centre of the heritage agenda of national and international organisations and researchers from all over the world. Of the various dimensions of sustainability identified, the sociocultural dimension has the most significant impact on safeguarding cultural heritage. In the approach to cultural heritage from the perspective of sustainability, evaluation processes through the use of indicators have broad advantages for formulating, analysing and monitoring cultural policies and management decision-making processes. In this context, a broad spectrum of proposals for sustainability indicators is applied from different approaches to various objects and contexts. However, in general, it is observed that most of the proposals formulate quantitative indicators with a substantial incidence of the economic and environmental dimensions, presenting limitations to address more sensitive aspects of cultural heritage, such as valuations and perceptions, which are not easily quantifiable due to their subjective nature. Therefore, this paper presents a methodological proposal for evaluating the sociocultural sustainability of urban heritage in medium-sized cities through a set of indicators formulated from a qualitative approach designed and tested in a medium-sized city in Argentina (Olavarría). The proposed methodology makes it possible to measure the progress and setbacks of heritage in pursuit of sustainability in the medium term. In addition, it facilitates the integration of evaluations from different disciplines. It incorporates the viewpoint of the main actors in heritage and heritage management, as well as making it possible to diagnose the situation of urban heritage in the areas of study.

Abstract The increasing prevalence of gut colonization with CTX-M extended-spectrum β-lactamase- and/or DHA plasmid-mediated AmpC-producing Escherichia coli is a concern. Here, we evaluate Nanopore-shotgun metagenomic sequencing (Nanopore-SMS) latest V14 chemistry to detect bla CTX-M and bla DHA genes from healthy stools. We test 25 paired samples characterized with culture-based methods (native and pre-enriched). Antimicrobial resistant genes (ARGs) are detected from reads and meta-assembled genomes (MAGs) to determine their associated genetic environments (AGEs). Sensitivity and specificity of native Nanopore-SMS are 61.1% and 100%, compared to 81.5% and 75% for pre-enriched Nanopore-SMS, respectively. Native Nanopore-SMS identifies only one sample with an AGE, whereas pre-enriched Nanopore-SMS recognizes 9/18 plasmids and 5/9 E. coli chromosomes. Pre-enriched Nanopore-SMS identifies more ARGs than native Nanopore-SMS (p

Abstract Chronic viral infections caused by the human immunodeficiency virus (HIV), hepatitis C (HCV), and hepatitis B (HBV) are common among patients with end-stage renal disease (ESKD). These infections were once considered contraindications to kidney transplantation due to potential risks associated with long-term immunosuppression. Improved management and antiviral therapies have changed the prognosis and survival of this group of patients, along with an increased experience in transplanting people with these viral infections. We report the first successful kidney transplant in an ESKD patient on hemodialysis with a history of concomitant HIV, HCV and HBV infection in Mexico.

Cranial operations are associated with a high risk of postoperative intracranial hemorrhage (pICH) and venous thromboembolic events, along with increased mortality and morbidity. With the use of acetylsalicylic acid (ASA) for prophylaxis becoming more prevalent, the risk of bleeding when ASA is administered preoperatively is unknown, as are the effects of discontinuation upon the occurrence of thromboembolic events, especially in societies with aging demographics. To address these questions, a retrospective analysis was performed using medical records and radiological images of 1862 patients subjected to brain tumor surgery over a decade in our department. The risk of pICH was compared in patients with metastases receiving ASA treatment versus patients not receiving ASA treatment. The occurrence of venous thromboembolic events after surgery was also evaluated. The study group consisted of 365 patients with different types of brain metastases. In total, 20 patients suffered pICH and 7 of these were associated with clinical neurological deterioration postoperatively. Of the 58 patients who took ASA preoperatively, 2 patients experienced pICH, compared with 5 patients in the non-ASA impact group (p = 0.120). Patients who took ASA were not at significantly higher risk of pICH and therefore a worse outcome compared to the group without ASA. Therefore, these data suggest that in patients at high cardiovascular risk, ASA can be safely continued during elective brain tumor surgery.

Abstract Restoring motor function after stroke necessitates involvement of numerous cognitive systems. However, the impact of damage to motor and cognitive network organization on recovery is not well understood. To discover correlates of successful recovery, we explored imaging characteristics in chronic stroke subjects by combining noninvasive brain stimulation and fMRI. Twenty stroke survivors (6 months or more after stroke) were randomly assigned to a single session of transcranial direct current stimulation (tDCS) or sham during image acquisition. Twenty healthy subjects were included as controls. tDCS was limited to 10 min at 2 mA to serve as a mode of network modulation rather than therapeutic delivery. Fugl-Meyer Assessments (FMA) revealed significant motor improvement in the chronic stroke group receiving active stimulation (p = 0.0005). Motor changes in this group were correlated in a data-driven fashion with imaging features, including functional connectivity (FC), surface-based morphometry, electric field modeling and network topology, focusing on relevant regions of interest. We observed stimulation-related changes in FC in supplementary motor (p = 0.0029), inferior frontal gyrus (p = 0.0058), and temporo-occipital (p = 0.0095) areas, though these were not directly related to motor improvement. The feature most strongly associated with FMA improvement in the chronic stroke cohort was graph topology of the dorsal attention network (DAN), one of the regions surveyed and one with direct connections to each of the areas with FC changes. Chronic stroke subjects with a greater degree of motor improvement had lower signal transmission cost through the DAN (p = 0.029). While the study was limited by a small stroke cohort with moderate severity and variable lesion location, these results nevertheless suggest a top-down role for higher order areas such as attention in helping to orchestrate the stroke recovery process.

Abstract Background Studies have shown that contamination of surfaces by illicit drugs frequently occurs in forensic laboratories when manipulating seized samples as well as in pharmacies and hospitals when preparing medicinal drugs. In this project, we extended these studies to a Drug Consumption Room to investigate drug levels and possible exposure of the staff members. Methods We investigated pre and post cleaning contamination by heroin and cocaine and their degradation products 6-monoacetylmorphine and benzoylecgonine on different surfaces (tables, counters, computers and door handles) and in the ambient air. We also collected urine and hair samples from staff members to check for potential short and long term contaminations. Results Medium to heavy contamination has been detected on most surfaces and door handles; as expected, air contamination was particularly high in the smoking room. Drug levels were

Abstract This study aimed to estimate the prevalence of asymptomatic and subpatent P. falciparum infections in the city of Bouaké, Central Côte d’Ivoire, to compare the performance of three tests, and to investigate potential P. falciparum histidine-rich protein 2 (pfhrp2) gene deletions. A cross-sectional survey was conducted in nine neighborhoods in Bouaké in 2016. Matched light microscopy (LM), rapid diagnostic test (RDT), and quantitative PCR (qPCR) data were used to determine the prevalence of P. falciparum infection and compare the performance of the three diagnostic tests. Pfhrp2/3 deletions were genotyped by digital PCR. Among 2313 individuals, 97.2% were asymptomatic and 2.8% were symptomatic. P. falciparum prevalence among symptomatic individuals was 25.8%, 30.3%, and 40.9% by LM, RDT, and varATS qPCR, respectively, and among asymptomatic individuals, it was 10.3%, 12.5%, and 34.9%. Asymptomatic infections comprised 96.4% of all malaria infections, with 58.2% detectable only by varATS qPCR. Although the prevalence of asymptomatic P. falciparum infections was higher in school-age children (5–14 years: 42.0%) compared to 10,000 parasites/µL. Individuals who were positive according to all three tests had significantly greater parasite density (856.8 parasites/µL; 95% CI 707.3–1,038) than did those who were positive by varATS qPCR only (13.7 parasites/µL; 95% CI 11.4–16.3) (p

Abstract Background Although the clinical signs of inflammatory breast cancer (IBC) resemble acute inflammation, the role played by infiltrating immune and stromal cells in this aggressive disease is uncharted. The tumor microenvironment (TME) presents molecular alterations, such as epimutations, prior to morphological abnormalities. These changes affect the distribution and the intricate communication between the TME components related to cancer prognosis and therapy response. Herein, we explored the global DNA methylation profile of IBC and surrounding tissues to estimate the microenvironment cellular composition and identify epigenetically dysregulated markers. Methods We used the HiTIMED algorithm to deconvolve the bulk DNA methylation data of 24 IBC and six surrounding non-tumoral tissues (SNT) (GSE238092) and determine their cellular composition. The prognostic relevance of cell types infiltrating IBC and their relationship with clinicopathological variables were investigated. CD34 (endothelial cell marker) and CD68 (macrophage marker) immunofluorescence staining was evaluated in an independent set of 17 IBC and 16 non-IBC samples. Results We found lower infiltration of endothelial, stromal, memory B, dendritic, and natural killer cells in IBC than in SNT samples. Higher endothelial cell (EC) and stromal cell content were related to better overall survival. EC proportions positively correlated with memory B and memory CD8+ T infiltration in IBC. Immune and EC markers exhibited distinct DNA methylation profiles between IBC and SNT samples, revealing hypermethylated regions mapped to six genes (CD40, CD34, EMCN, HLA-G, PDPN, and TEK). We identified significantly higher CD34 and CD68 protein expression in IBC compared to non-IBC. Conclusions Our findings underscored cell subsets that distinguished patients with better survival and dysregulated markers potentially actionable through combinations of immunotherapy and epigenetic drugs.

Abstract Objective To assess seizure and developmental outcomes, their predictors, and complications in 160 children who, between 1998 and 2022, underwent surgery for lesional epilepsy with curative intent before the age of 3 years. To compare trends in epilepsy surgery in this age group before and after the year 2014. Methods Retrospective multicenter study. Descriptive and univariate analyses, and multivariable models for all outcomes. Results These 160 patients (76 F; 47.5%) underwent 169 surgeries (age at surgery 20.4 ± 9.4 months). At the last follow‐up (77 ± 57.4 months), 121 patients (75.6%) were in Engel class I, 106 (66.2%) of whom were in Engel class Ia. Antiseizure medications were stopped in 84 patients (52.5%). Complications requiring reoperations were observed in 16 patients (10%; 9.5% of surgeries) and unexpected permanent deficits in 12 (7.5%; 7.1% of surgeries). Postoperative cognitive functions remained unchanged in 56 patients (44.4%), improved in 51 (40.5%), and worsened in 19 (15.1%). Multivariable analyses showed that the probability of achieving Engel class Ia was lower when the duration of epilepsy was longer, patients underwent preoperative video‐EEG, and unexpected postoperative permanent deficits occurred. Cognitive improvement after surgery was associated with lower preoperative seizure frequency, better preoperative developmental level, and a longer postoperative follow‐up. FCDII and tumors were the histopathologies carrying a higher probability of achieving seizure freedom, while polymicrogyria was associated with a lower probability of cognitive improvement. The number of patients operated on after 2014 was higher than before (61.3% vs. 38.7%), with stable outcomes. Significance Epilepsy surgery is effective and safe in infants and toddlers, although the complication rate is higher than seen in older patients. Shorter duration of epilepsy, lower seizure frequency, no need for video‐EEG, tumors, and some malformations of cortical development are robust predictors of seizure and cognitive outcome that may be exploited to increase earlier referral. Plain Language Summary This study analyzed the results of epilepsy surgery in 160 children who had been operated on before the age of 3 years at four Italian centers between 1998 and 2022. At the last follow‐up (77 ± 57.4 months), 121 patients (75.6%) were free from disabling seizures, of which 106 (66.2%) were completely seizure‐free since surgery. Major surgical complications occurred in 28 patients (17.5%), which is higher than observed with epilepsy surgery in general, but similar to hemispheric/multilobar surgery. Postoperative cognitive function remained unchanged in 56 patients (44.4%), improved in 51 (40.5%), and worsened in 19 (15.1%). Epilepsy surgery is effective and safe in infants and toddlers.

Sexual arousal (SA) is a multidimensional experience that includes cognitive, emotional, motivational and physiological components. Texts with erotic content have been used to generate a state of SA. Erotic texts often include aggressive content that has not been evaluated in relation to SA. The aim of this work was to compare cortical functionality in women when reading a sexually explicit text (SET) and a sexually explicit text with aggression (SETA). Twenty-seven women participated. The EEG activity of the frontal, temporal and parietal locations was recorded during the reading of both texts. The participants found the SET to be more pleasant than the SETA. Both texts were identified as triggers of general and SA. While reading the SETA, there was an increase in absolute power in the frontal and parietal locations, a higher intrahemispheric correlation between the left frontal and temporal locations in fast frequency bands and a greater interhemispheric correlation between the frontal locations in the delta and alpha1 bands. These findings indicate that cortical functionality during SA in women differs based on the content and context of the erotic material being read, possibly associated with mechanisms that underlie the processing and incentive value assignment of stimuli with sexual and aggressive connotations.

Abstract Background Hox proteins interact with DNA and many other proteins, co-factors, transcriptional factors, chromatin remodeling components, non-coding RNAs and even the extracellular matrix that assembles the Hox complexes. The number of interacting partners continues to grow with diverse components and more transcriptional factors than initially thought. Hox complexes present many activities, but their molecular mechanisms to modulate their target genes remain unsolved. Results In this paper we showed the protein-protein interaction of Antp with Ubx through the homeodomain using BiFC in Drosophila. Analysis of Antp-deletional mutants showed that AntpHD helixes 1 and 2 are required for the interaction with Ubx. Also, we found a novel interaction of Ubx with TBP, in which the PolyQ domain of TBP is required for the interaction. Moreover, we also detected the formation of two new trimeric complexes of Antp with Ubx, TBP and Exd using BiFC-FRET; these proteins, however, do not form a trimeric interaction with BIP2 or TFIIEβ. The novel trimeric complexes reduced Antp transcriptional activity, indicating that they could confer specificity for repression. Conclusions Our results increase the number of transcriptional factors in the Antp and Ubx interactomes that form two novel trimeric complexes with TBP and Exd. We also report a new Ubx interaction with TBP. These novel interactions provide important clues of the dynamics of Hox-interacting complexes involved in transcriptional regulation, contributing to better understand Hox function.

New Italian data on the distribution of the Annex I Habitats are reported in this contribution. Specifically, 9 new occurrences in Natura 2000 sites are presented and 34 new cells are added in the EEA 10 km × 10 km reference grid. The new data refer to the Italian administrative regions of Abruzzo, Apulia, Calabria, Latium, Lombardy, Marche, Sardinia, Sicily, Tuscany and Umbria.

Abstract Immune cell-derived extracellular vesicles (EV) affect tumor progression and hold promise for therapeutic applications. Eosinophils are major effectors in Th2-related pathologies recently implied in cancer. Here, we evaluated the anti-tumor activities of eosinophil-derived EV following activation with the alarmin IL-33. We demonstrate that IL-33-activated mouse and human eosinophils produce higher quantities of EV with respect to eosinophils stimulated with IL-5. Following incorporation of EV from IL-33-activated eosinophils (Eo33-EV), but not EV from IL-5-treated eosinophils (Eo5-EV), mouse and human tumor cells increased the expression of cyclin-dependent kinase inhibitor (CDKI)-related genes resulting in cell cycle arrest in G0/G1, reduced proliferation and inhibited tumor spheroid formation. Moreover, tumor cells incorporating Eo33-EV acquired an epithelial-like phenotype characterized by E-Cadherin up-regulation, N-Cadherin downregulation, reduced cell elongation and migratory extent in vitro, and impaired capacity to metastasize to lungs when injected in syngeneic mice. RNA sequencing revealed distinct mRNA signatures in Eo33-EV and Eo5-EV with increased presence of tumor suppressor genes and enrichment in pathways related to epithelial phenotypes and negative regulation of cellular processes in Eo33-EV compared to Eo5-EV. Our studies underscore novel IL-33-stimulated anticancer activities of eosinophils through EV-mediated reprogramming of tumor cells opening perspectives on the use of eosinophil-derived EV in cancer therapy.

Abstract Background In the fight against GBM, drug repurposing emerges as a viable and time-saving approach to explore new treatment options. Chlorpromazine, an old antipsychotic medication, has recently arisen as a promising candidate for repositioning in GBM therapy in addition to temozolomide, the first-line standard of care. We previously demonstrated the antitumor efficacy of chlorpromazine and its synergistic effects with temozolomide in suppressing GBM cell malignant features in vitro. This prompted us to accomplish a Phase II clinical trial to evaluate the efficacy and safety of adding chlorpromazine to temozolomide in GBM patients with unmethylated MGMT gene promoter. In this in vitro study, we investigate the potential role of chlorpromazine in overcoming temozolomide resistance. Methods In our experimental set, we analyzed Connexin-43 expression at both the transcriptional and protein levels in control- and chlorpromazine-treated GBM cells. DNA damage and subsequent repair were assessed by immunofluorescence of γ-H2AX and Reverse-Phase Protein microArrays in chlorpromazine treated GBM cell lines. To elucidate the relationship between DNA repair systems and chemoresistance, we analyzed a signature of DNA repair genes in GBM cells after treatment with chlorpromazine, temozolomide and Connexin-43 downregulation. Results Chlorpromazine treatment significantly downregulated connexin-43 expression in GBM cells, consequently compromising connexin-dependent cellular resilience, and ultimately contributing to cell death. In line with this, we observed concordant post-translational modifications of molecular determinants involved in DNA damage and repair pathways. Our evaluation of DNA repair genes revealed that temozolomide elicited an increase, while chlorpromazine, as well as connexin-43 silencing, a decrease in DNA repair gene expression in GBM cells. Conclusions Chlorpromazine potentiates the cytotoxic effects of the alkylating agent temozolomide through a mechanism involving downregulation of Cx43 expression and disruption of the cell cycle arrest essential for DNA repair processes. This finding suggests that chlorpromazine may be a potential therapeutic strategy to overcome TMZ resistance in GBM cells by inhibiting their DNA repair mechanisms.

Abstract Gulf War Illness (GWI) is characterized by a wide range of symptoms that manifests largely as gastrointestinal symptoms. Among these gastrointestinal symptoms, motility disorders are highly prevalent, presenting as chronic constipation, stomach pain, indigestion, diarrhea, and other conditions that severely impact the quality of life of GWI veterans. However, despite a high prevalence of gastrointestinal impairments among these veterans, most research attention has focused on neurological disturbances. This perspective provides a comprehensive overview of current in vivo research advancements elucidating the underlying mechanisms contributing to gastrointestinal disorders in GWI. Generally, these in vivo and in vitro models propose that neuroinflammation alters gut motility and drives the gastrointestinal symptoms reported in GWI. Additionally, this perspective highlights the potential and challenges of in vitro bioengineering models, which could be a crucial contributor to understanding and treating the pathology of gastrointestinal related-GWI.

In recent years, K-Pop has become a global sensation, especially in Indonesia, where it has a massive fan base on social media. While K-Pop brings joy to many, it has also raised concerns about its impact on body image, especially among young fans known as K-Popers. This study explores how young women who are fans of K-Pop feel about their bodies. Through interviews and observations, three participants—Yerin, Joy, and Rose—shared their thoughts. Despite K-Pop's influence on beauty standards, these women have a positive body image. They feel good about themselves and focus on self-confidence rather than what others think. Their stories show how they navigate societal pressures and find acceptance for their bodies. By prioritizing self-esteem and support from friends, they maintain a positive outlook despite external beauty ideals. This research sheds light on how K-Pop fans view their bodies and offers insights into promoting body positivity. It's important for researchers and others to continue exploring these issues to create a more accepting environment for everyone to feel good about themselves.

Abstract Background Inflammatory breast cancer (IBC) is a rare disease characterized by rapid progression, early metastasis, and a high mortality rate. Methods Genome-wide DNA methylation analysis (EPIC BeadChip platform, Illumina) and somatic gene variants (105 cancer-related genes) were performed in 24 IBCs selected from a cohort of 140 cases. Results We identified 46,908 DMPs (differentially methylated positions) (66% hypomethylated); CpG islands were predominantly hypermethylated (39.9%). Unsupervised clustering analysis revealed three clusters of DMPs characterized by an enrichment of specific gene mutations and hormone receptor status. The comparison among DNA methylation findings and external datasets (TCGA-BRCA stages III-IV) resulted in 385 shared DMPs mapped in 333 genes (264 hypermethylated). 151 DMPs were associated with 110 genes previously detected as differentially expressed in IBC (GSE45581), and 68 DMPs were negatively correlated with gene expression. We also identified 4369 DMRs (differentially methylated regions) mapped on known genes (2392 hypomethylated). BCAT1, CXCL12, and TBX15 loci were selected and evaluated by bisulfite pyrosequencing in 31 IBC samples. BCAT1 and TBX15 had higher methylation levels in triple-negative compared to non-triple-negative, while CXCL12 had lower methylation levels in triple-negative than non-triple-negative IBC cases. TBX15 methylation level was associated with obesity. Conclusions Our findings revealed a heterogeneous DNA methylation profile with potentially functional DMPs and DMRs. The DNA methylation data provided valuable insights for prognostic stratification and therapy selection to improve patient outcomes. Graphical Abstract

Abstract Experimental autoimmune encephalomyelitis (EAE) is a demyelinating disease affecting the central nervous system (CNS) in animals that parallels several clinical and molecular traits of multiple sclerosis in humans. Herpes simplex virus type 1 (HSV-1) infection mainly causes cold sores and eye diseases, yet eventually, it can also reach the CNS, leading to acute encephalitis. Notably, a significant proportion of healthy individuals are likely to have asymptomatic HSV-1 brain infection with chronic brain inflammation due to persistent latent infection in neurons. Because cellular senescence is suggested as a potential factor contributing to the development of various neurodegenerative disorders, including multiple sclerosis, and viral infections may induce a premature senescence state in the CNS, potentially increasing susceptibility to such disorders, here we examine the presence of senescence-related markers in the brains and spinal cords of mice with asymptomatic HSV-1 brain infection, EAE, and both conditions. Across all scenarios, we find a significant increases of senescence biomarkers in the CNS with some differences depending on the analyzed group. Notably, some senescence biomarkers are exclusively observed in mice with the combined conditions. These results indicate that asymptomatic HSV-1 brain infection and EAE associate with a significant expression of senescence biomarkers in the CNS.

Abstract Objectives Genomic surveillance of Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) is crucial for virulence, drug-resistance monitoring, and outbreak containment. Methods Genomic analysis on 87 KPC-Kp strains isolated from 3 Northern Italy hospitals in 2019-2021 was performed by whole genome sequencing (WGS), to characterize resistome, virulome, and mobilome, and to assess potential associations with phenotype resistance and clinical presentation. Maximum Likelihood and Minimum Spanning Trees were used to determine strain correlations and identify potential transmission clusters. Results Overall, 15 different STs were found; the predominant ones included ST307 (35, 40.2%), ST512/1519 (15, 17.2%), ST20 (12, 13.8%), and ST101 (7, 8.1%). 33 (37.9%) KPC-Kp strains were noticed to be in five transmission clusters (median number of isolates in each cluster: 5 [3-10]), four of them characterized by intra-hospital transmission. All 87 strains harbored Tn4401a transposon, carrying bla KPC-3 (48, 55.2%), bla KPC-2 (38, 43.7%), and in one case (1.2%) bla KPC-33, the latter gene conferred resistance to ceftazidime/avibactam (CZA). Thirty strains (34.5%) harbored porin mutations; of them, 7 (8.1%) carried multiple Tn4401a copies. These strains were characterized by significantly higher CZA minimum inhibitory concentration compared with strains with no porin mutations or single Tn4401a copy, respectively, even if they did not overcome the resistance breakpoint of 8 ug/mL. Median 2 (IQR:1-2) virulence factors per strain were detected. The lowest number was observed in ST20 compared to the other STs (p

We studied the vascular flora of the isthmus of Feniglia, a Nature Reserve in southern Tuscany stretching between the Italian Peninsula and Mt. Argentario. Since the nineteenth century, the area has undergone significant environmental modifications due to intensive grazing and deforestation. Later, rehabilitation interventions were carried out, including reforestation and planting of dune species, making it an interesting protected area to study for its botanical aspects. Therefore, we aim to compile the floristic inventory of this distinctive Mediterranean area by integrating old and new data. The checklist comprises 502 specific and subspecific taxa of vascular plants. The life-form spectrum shows a predominance of therophytes, followed by hemicryptophytes. The chorological spectrum highlights the dominance of Mediterranean species, followed by Euromediterranean and Eurasian species. The presence of 15 species of regional importance, 3 species of the Italian red list and 3 Italian endemics (Ornithogalum exscapum Ten., Limonium multiforme Pignatti, Linaria purpurea (L.) Mill.) is noteworthy. Despite the environmental changes undergone in this area, our study reports the presence of remarkable species, including rare ones such as Atriplex littoralis L., Ruppia spiralis L. ex Dumort., and species at their distribution margin in the Italian Peninsula, such as Staphisagria macrosperma Spach, Juncus sorrentinoi Parl. and Maresia nana (DC.) Batt. Additionally the presence of 26 alien species, of which 15 are invasive, 9 naturalized and 2 casual, indicates a relatively low presence of alien species. Notwithstanding the significant environmental changes that the Feniglia isthmus has experienced in the last century and the high tourist pressure during the summer, the presence of the protected area suggested the effectiveness and benefits of such a form of protection for plant diversity.

Objetivos: Evaluar el efecto de extractos cítricos sobre la adhesión C. albicans a acrílicos de uso en odontología. Métodos: Extractos cítricos: se secaron y pulverizaron cáscaras de limón y naranja, sometiéndolas a maceración a 50°C, solución etanol/agua 50:50 y relación masa/solvente 1:5. Discos de acrílico: se prepararon discos de espesor de 2 mm, diámetro de 2,5cm y superficies pulidas según indicaciones del fabricante. Microorganismos: suspensión de C. albicans (CA), ATCC 5314, a 0,5 Mc Farland. Prueba de adhesión: se incubaron los discos de acrílico con diferentes tratamientos: 1: CA + extracto de limón (0,8 g/mL); 2: CA + extracto de naranja (0,6 g/mL); y 3: control positivo CA sin tratamiento. Las muestras se incubaron a 37°C/24hs. Los discos se lavaron con PBS y fueron metalizadas para su análisis por microscopía electrónica de barrido. Se realizó recuento de levaduras y formas filamentosas (unidades/campo) e identificación morfológica digital a 5 kV con un aumento de X-750 en instrumento SEM. Resultados: Se observó inhibición en la adhesión de levaduras con los tratamientos con extractos cítricos. Se registraron un 79% y 60% menos de levaduras en discos sometidos a tratamiento 1 y 2 respectivamente. En relación a la morfología, no se observaron hifas/tubos germinativos y, se mantuvo la estructura levaduriforme y tamaño promedio. Conclusiones: El extracto de limón fue el de mayor efectividad sobre la adhesión del hongo a acrílicos de uso odontológico, revelando la potencial utilidad de compuestos naturales para la formulación de soluciones de limpieza de prótesis dentales.

Conflict of interest: No Introduction and Objectives: Background: Severe alcohol-associated hepatitis (AH) is considered a common precipitant of acute-on-chronic liver failure (ACLF). This study aims to characterize the association between AH and ACLF, focusing on mortality across different ACLF grades. Patients / Materials and Methods: Multicenter prospective cohort study. We included patients admitted with severe AH between 2015–2022. The main outcome was mortality by ACLF grade during admission. The analysis included survival analysis using Cox regression. We adjusted multivariable models based on the main predictors of mortality observed in prior studies. Results and Discussion: We prospectively included 646 patients from 24 centers and 8 countries. Age 49.9±11.7 years, 85.1% of men and 64.4% had a previous diagnosis of cirrhosis. Median MELD at admission was 25 [20-31] points, 46.5% of patients were treated with corticosteroids, and only 2.2% underwent liver transplantation (LT). Around 67.4% of patients fulfilled ACLF criteria: 10.1% grade 1, 19.2% grade 2, and 38.1% grade 3. The most frequent organ dysfunctions were 76.2% liver, 40.8% brain, 43.2% coagulation, 29.6% renal, 29.4% circulatory, and 18.9% lung failure. Survival at 180 days was 77.8% (95%CI: 72.1-82.6%) in those without ACLF grade 3 and 28.0% (95%CI: 20.5–36.0%) in those with ACLF grade 3 (p

Conflict of interest: No Introduction and Objectives: Primary biliary cholangitis (PBC) may present differently depending on various factors such as ethnicity and genetic background. Latin America has a highly admixed population with a unique genetic diversity compared to other regions of the world. However, there is limited information available on the presentation and epidemiology of PBC in this region. This study aims to address the epidemiology of PBC in Latin America. Patients / Materials and Methods: Ongoing retrospective, international, multicentric cohort study sponsored by ALEH that enrolls PBC patients from different countries in Latin America. Results and Discussion: Data were accrued on 231 patients [Brazil (52%), Argentina (27.4%), Chile (10.8%), Costa Rica (4.5%), Cuba (3.6%), and Mexico (0.9%)], 92.1% female (mean age at diagnosis 50.5 years), 25.6% with cirrhosis at baseline. Overlap with autoimmune hepatitis was reported in 16.0% of cases. Most patients were symptomatic (67.9%) at diagnosis, with fatigue (41.9%) and pruritus (40.5%) being the main symptoms. Anti-mitochondrial antibodies (AMA) were positive in 70.8% and antinuclear antibodies (ANA) in 60.6%. Hashimoto thyroiditis (23.7%) and Sjogren syndrome (9.1%) were the most common extrahepatic autoimmune diseases associated with PBC. Mean baseline alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, and bilirubin levels were 445.9 (± 407), 89.8 (± 137.2), 37.6 (± 8.9) U/L, and 1.6 (± 3.1) mg/dL, respectively. Almost all patients (99.1%) were treated with ursodeoxycholic acid (UDCA). 67.4% achieved adequate response to UDCA according to the Toronto criteria and 32% normalized alkaline phosphatase at 12 months. Only 19.9% received second-line therapy, all with fibrates (89.1% bezafibrate, 8.7% ciprofibrate, 4.3% fenofibrate). Of the patients, 9% died, with 33% of deaths being liver-related, while 6% underwent liver transplantation. Hepatocellular carcinoma was diagnosed in 1.7% of patients. Conclusions: In this unprecedented study, the epidemiology of PBC in Latin America appears similar to that in other parts of the world. However, lower rates of AMA positivity were observed, and most patients were still diagnosed with symptomatic disease. Second-line therapy options were limited to the availability of fibrates only.