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IntroductionThe aim of this study was to evaluate the fit performance of implant infrastructures manufactured by five different techniques: heat-press (IPS), milling (ZIR), lost wax (CER), calcinable cylinder (CAL), and CAD/Waxx® (CAD).MethodsThe methodology was based on the Replica Technique, which can simulate and evaluate the fit of the infrastructure on the implant component. Thus, each infrastructure was internally filled with low-viscosity silicone addition and seated on the component until its final setting, obtaining the replica of the cementation space. After removing the coping containing the silicone film, light-density silicone was inserted addition in place of the components, and in its surroundings, condensation silicone was applied, establishing support for the assembly. The joint was sectioned mesiodistally, photographed, and analyzed in image processing software in order to measure the thickness of the interface infrastructure/implant at five different areas: marginal opening (M), gingival-axial angle (G-A), axial region (A), axial-occlusal angle (A-O) and occlusal surface (O).ResultsThe lowest and the highest average thickness between groups was, respectively, IPS: 187.5 μm and CAD: 221.6 μm, with statistically significant differences (p

IntroductionGalvanic vestibular stimulation (GVS) is a simple, safe, and noninvasive method of neurostimulation that can be used to improve body balance. Several central nervous system diseases cause alterations in body balance, including HTLV-1-associated myelopathy (HAM).ObjectiveTo test GVS as a balance rehabilitation strategy for HAM.MethodsThis study is a quasi-experimental clinical trial in which postural balance was compared before and after a GVS rehabilitation protocol applied to 20 patients with HAM, 12 women and 8 men, average age of 78 and 79 years, respectively. They were followed for nine months after the end of the GVS protocol, which consisted of one GVS session per week for 12 consecutive weeks. The GVS current intensity was progressively increased from 1.0 milliamperes (mA) to 3.5 mA until the third session and maintained at 3.5 mA until the 12th session. The electrical stimulation time progressively increased from 9 min in the first session to 18 min in the second session and maintained at 30 min from the third session onwards. Postural balance was assessed by Time up and go test (TUG), Berg balance scale (BBS) and posturography that were performed before the beginning of the intervention, during the intervention (6th week), at the end of the intervention (12th week) and after 9 months of follow-up without electrical stimulation.ResultsIn a blind comparison, in the 12th week of stimulation, improvement was observed in all the tests. In TUG, time in seconds changed from 28 before to 18 after GVS (p

Type 1 diabetes (T1D) is characterized by a prolonged autoimmune attack resulting in the massive loss of insulin-producing beta cells. The initiation and progression of T1D depends on a complex interaction between genetic, immunological and environmental factors. Epidemiological, experimental and clinical evidence suggest a link between viral infections, particularly Coxsackievirus type B (CVB), and T1D development. Specifically, infections by the CVB serotype 1 (CVB1) contribute to the triggering of autoimmunity against beta cells in genetically predisposed individuals, and prolonged and probably non-lytic infections by CVB are associated with the development of T1D. However, the molecular mechanisms underlying CVB1 replication and establishing persistent infections in human pancreatic beta cells remain poorly understood. Here we show that the N6-methyladenosine (m6A) RNA epigenetic modification machinery regulates CVB1 amplification in the human beta cells. Using small interfering RNA (siRNA) targeting m6A writers and erasers, we observed that downregulation of m6A writers increases CVB1 amplification, while the downregulation of m6A erasers decreases it. Notably, the inhibition of Fat Mass and Obesity-associated protein (FTO), a key m6A eraser, reduced by 95% the production of infectious CVB1 in both human insulin-producing EndoC-βH1 cells and in induced pluripotent stem cell (iPSC)-derived islets. The FTO inhibitor reduced CVB1 expression within 6 h post-infection, suggesting a direct regulation of the CVB1 genome by m6A modification. Furthermore, in the absence of viral replication, FTO inhibition also decreased the translation of the incoming CVB1 genome, indicating that m6A plays a critical role in the initial stages of viral RNA translation. In addition, modulation of the m6A machinery affected the type I interferon response after poly-IC transfection, a mimic of RNA virus replication, but did not affect the cellular antiviral response in CVB1-infected cells. Altogether, these observations suggest that m6A directly affects CVB1 production. Our study provides the first evidence that the m6A epigenetic modification machinery controls CVB amplification in human pancreatic beta cells. This suggests that the m6A machinery is a potential target to control CVB infection in T1D and raises the possibility of an epigenetic control in the establishment of persistent CVB infections observed in the pancreas in individuals with type 1 diabetes.

IntroductionThe prevalence of germline pathogenic/likely pathogenic variants (P/LP) in high and moderate penetrance (HMP) genes is approximately 7%–10% among breast cancer (BC) patients. The prevalence and spectrum of BC P/LP variants are affected by several factors. There are limited genetic data from Brazilian patients with BC.MethodsThis is a retrospective cross-sectional study that aims to evaluate the germline profile of P/LP variants in 13 HMP BC genes (BRCA1, BRCA2, PALB2, TP53, CDH1, NF1, PTEN, STK11, CHEK2, ATM, BARD1, RAD51C, and RAD51D) in patients diagnosed with BC in Brazil. All patients were tested using multigene NGS panels covering from 35 to 105 genes. Primary endpoint was the prevalence of P/LP variants in BRCA1/2 and in other HMP genes. Secondary endpoints were stratified analyses according to age and BC subtype.ResultsThis cohort involved 2,208 patients with BC from 2019 to 2023. Most patients (79.7%) were from Southeastern Brazil. The median age at genetic testing was 47 years, and most patients (59.4%) were ≤50 years. The BC subtype was available in 641 cases: 264 patients (41.2%) were HR+/HER2−, 116 (18.1%) were HER2+, and 261 (40.7%) had triple-negative breast cancer (TNBC). Overall, 215 (9.7%) had a P/LP in HMP genes, including 5.8% in BRCA1/2. The most frequent variants were found in BRCA2, BRCA1, and TP53. The founder variant R337H accounted for 79% of all TP53 pathogenic variants, representing 1% of the overall population. Deleterious variants in BRCA1/2 were more common in patients ≤50 years (7.7%) and TNBC (10.7%). In other HMP BC genes, the prevalence of P/LP variants did not significantly vary according to age and BC molecular subtype. The overall VUS rate in HMP genes was 19.6%.ConclusionIn Brazil, the epidemiology of deleterious variants in HMP is comparable to published US and EU cohorts. The Brazilian TP53 R337H is a prevalent variant in BC patients. Deleterious BRCA1/2 variants vary according to age and BC subtype. Our study gives a broader understanding of BC risk genes and has opened doors to optimized testing and surveillance strategies in Brazil.

Spinal cord injury (SCI) can cause significant motor, sensory, and autonomic dysfunction by disrupting neural connections. As a result, it is a global health challenge that requires innovative interventions to improve outcomes. This review assesses the wide-ranging impacts of SCI and focuses on the laparoscopic implantation of neuroprosthesis (LION) as an emerging and promising rehabilitation technique. The LION technique involves the surgical implantation of electrodes on lumbosacral nerves to stimulate paralyzed muscles. Recent findings have demonstrated significant improvements in mobility, sexual function, and bladder/bowel control in chronic SCI patients following LION therapy. This manuscript revisits the potential physiological mechanisms underlying these results, including neuroplasticity and modulation of autonomic activity. Additionally, we discuss potential future applications and amendments of LION therapy. This study emphasizes the potential of neuromodulation as a complementary approach to traditional rehabilitation, that can provide a beacon of hope for improving functionality and quality of life for individuals with SCI.

Pathogenic Leptospira are spirochetes that cause leptospirosis, a worldwide zoonotic disease. Leptospirosis affects humans and animals, with approximately 1 million human infections and 60,000 deaths per year. The diversity of leptospiral strains and serovars allied to the fact that pathogenesis is not yet fully understood, make the development of an effective vaccine against leptospirosis a challenge. Outer membrane and secreted proteins are considered potential antigens since they play a vital role in mediating interactions with host molecules. Several domains or motifs have been reported to participate in the leptospiral infection process. Among them, leucine-rich repeat (LRR) proteins have been highlighted as attractive multipurpose proteins, exhibiting a broad spectrum of ligands and having a putative role in bacterial pathogenesis. Indeed, genome annotation of leptospiral species pointed out that LRR proteins are predominant in pathogenic strains, a feature that corroborates this hypothesis. A few LRR proteins of L. santarosai, L. borgpetersenii and L. interrogans have been studied and their possible role in virulence was proposed. Yet, a mechanistic and broad investigation of LRR proteins was not fully performed. In this review, a comprehensive in silico analysis of 21 LRR proteins of L. interrogans was performed in relation to structure, function, dynamics and virulent potential that will contribute to understanding the key role of these domains in the underlying mechanisms of leptospiral infection.

Metarhizium is widely known for its role as an arthropod biocontrol agent and plant bioinoculant. By using mass-production industrial methods, it is possible to produce large amounts of fungal single-celled propagules (including blastospores) to be applied in the field. However, in the environment, the solar ultraviolet components (particularly UV-B) can harm the fungus, negatively impacting its pathogenicity toward the arthropod pest. The present study is the first to use comparative genome-wide transcriptome analyses to unveil changes in gene expression between Metarhizium pingshaense blastospores exposed or not to UV-B. Relative blastospores culturability was calculated 72 h after UV-B exposure and exhibited 100% culturability. In total, 6.57% (n = 728) out of 11,076 predicted genes in M. pingshaense were differentially expressed after UV-B exposure: 320 genes (44%; 320/728) were upregulated and 408 (56%; 408/720) were downregulated in the UV-B exposed blastospores. Results unveiled differentially expressed gene sets related to fungal virulence, production of secondary metabolites, and DNA repair associated with UV damage; genes related to virulence factors were downregulated, and genes associated with nucleotide excision repair were upregulated. These findings illustrate critical aspects of Metarhizium blastospores strategies to overcome UV-B damage and survive solar radiation exposures in insulated fields.

Enteropathogenic Escherichia coli (EPEC) strains are subdivided into typical (tEPEC) and atypical (aEPEC) according to the presence or absence of a virulence-associated plasmid called pEAF. Our research group has previously demonstrated that two aEPEC strains, 0421-1 and 3991-1, induce an increase in mucus production in a rabbit ileal loop model in vivo. This phenomenon was not observed with a tEPEC prototype strain. Few studies on aEPEC strains evaluating their capacity to induce intestinal mucus hypersecretion were done. This study aimed to investigate aEPEC strains regarding their genotypic and phenotypic characteristics, their ability to alter mucus production in an in vivo intestinal infection model, and their potential mucinolytic activity. To investigate the relationship between strains 0421-1 and 3991-1 and 11 other aEPEC strains, their serotypes, sequence types (ST), and virulence factors (VF), several sequencing and genomic analyses were carried out. The study also involved researching the reproduction of mucus hypersecretion in rabbits in vivo. We found that the two mucus-inducing strains and two other strains (1582-4 and 2531-13) shared the same phylogroup (A), ST (378), serotype (O101/O162:H33), and intimin subtype (ι2), were phylogenetically related, and induced mucus hypersecretion in vivo. A wide diversity of VFs was found among the strains, confirming their genomic heterogeneity. However, among the genes studied, no unique virulence factor or gene set was identified exclusively in the mucus-inducing strains, suggesting the multifactorial nature of this phenomenon. The two strains (1582-4 and 2531-13) closely related to the two aEPEC strains that induced mucus production in vivo also induced the phenomenon. The investigation of the mucinolytic activity revealed that all aEPEC strains used mucins as their carbon sources. Ten of the 13 aEPEC strains could cross a mucin layer, and only four adhered better to agar containing mucin than to agar without mucin. The present study paves the way for subsequent investigations into the molecular mechanisms regarding cellular interactions and responses, as well as the correlation between virulence factors and the induction of mucus production/expression during aEPEC infections.

The study targeted to compare the nutritional profile of two varieties of açaí, the white and purple, found in different mesoregions of Pará, Brazil. The research focused on analyzing levels of total phenolics, total anthocyanins, antioxidant capacity, and mineral composition in these two varieties. The study sought to identify significant differences between the two varieties in terms of nutritional composition and antioxidant potential, providing valuable information into the specific nutritional and functional properties of each type of açaí studied. Higher levels of total phenolics, total anthocyanins, and antioxidant capacity were observed in purple açaí fruits, with values of 806.17 ± 17.48 mgGAE/100 g, 81.73 ± 1.77 mg/100 g, and 19.25 ± 0.35 μmol of Trolox equivalent (TE)/g, respectively, compared to 401.92 ± 52.70 mgGAE/100 g, 37.70 ± 5.34 mg/100 g, and 6.17 ± 1.07 μmol TE/g in white açaí. HPLC-MS analysis identified and quantified monomeric anthocyanins in white açaí, using two distinct analytical methods, revealing average values of 0.29 and 1.05 μg/100 g for cyanidin-3-glucoside and between 0.74 and 3.13 μg/100 g for cyanidin-3-rutinoside, respectively, which were higher than those found in yellow tropical fruits. The quality of both purple and white açaí varied significantly among mesoregions, with fruits from floodplain soils demonstrating superior quality compared to those from sandy and solid soils in southeastern Pará. Mineral composition and microbiological characteristics were similar between white and purple açaí. These findings underscore the significant influence of mesoregion and soil type on açaí quality, emphasizing the superiority of fruits grown in floodplain soils.

Despite extensive efforts to unravel tumor behavior and develop anticancer therapies, most treatments fail when advanced to clinical trials. The main challenge in cancer research has been the absence of predictive cancer models, accurately mimicking the tumoral processes and response to treatments. The tumor microenvironment (TME) shows several human-specific physical and chemical properties, which cannot be fully recapitulated by the conventional 2D cell cultures or the in vivo animal models. These limitations have driven the development of novel in vitro cancer models, that get one step closer to the typical features of in vivo systems while showing better species relevance. This review introduces the main considerations required for developing and exploiting tumor spheroids and organoids as cancer models. We also detailed their applications in drug screening and personalized medicine. Further, we show the transition of these models into novel microfluidic platforms, for improved control over physiological parameters and high-throughput screening. 3D culture models have provided key insights into tumor biology, more closely resembling the in vivo TME and tumor characteristics, while enabling the development of more reliable and precise anticancer therapies.

Pathogenic spirochaetes of the genus Leptospira are the etiological agents of leptospirosis, a zoonotic infection worldwide. The disease is considered an emerging and re-emerging threat due to global warming, followed by heavy rainfall and flooding when outbreaks of leptospirosis occur. Adhesion to host tissues is mediated by surface/extracellular proteins expressed by pathogens during infection. Leucine-rich repeat (LRR) domain-containing proteins seem to be important for the virulence of pathogenic Leptospira and their role has been recently examined. Here, we report the characterization of two LRR-proteins encoded by LIC11051 and LIC11505. They present 7 and 17 LRR motifs, respectively. LIC11051 was found mainly in the P1 subclade, whereas LIC11505 was identified with higher identity in subclade P1, but was also found in subclades P2, S1, and S2. The recombinant proteins were recognized by antibodies in leptospirosis serum samples, suggesting their expression during infection. rLIC11505 contains a broad spectrum of ligands, including GAG and integrin receptors, whereas rLIC11051 showed limited binding activity. The attachment of proteins to ligands was specific, dose-dependent, and saturable. Compared to their role in adhesion, both proteins were shown to be secreted, with the ability to reassociate with the bacteria. Taken together, our data suggested that LIC11051 and LIC11505 participate in leptospiral pathogenesis. To the best of our knowledge, this is the first report showing leptospiral LRR-proteins exhibiting GAG and integrin receptor-binding properties.

IntroductionThe aim of this study was to compare the effectiveness of brucellosis control and reproductive performance using one or two doses of RB51 strain vaccine.MethodsThis experiment was conducted at two commercial beef farms (A, n = 477; and B, n = 673), which were selected due to their records of endemic brucellosis and a prevalence of 6 to 8% of seropositive cows. An initial serology screening (Day 0) was carried out in all cows using the Rose Bengal test (RB), and presumptive positive results were re-evaluated using a slow agglutination test with 2-mercaptoethanol (2-ME). Seropositive cows (64, 5.6%) were culled. Non-reactive cows were then randomly assigned into three experimental groups: G1, a single vaccination with RB51; G2, two doses of RB51 given 6 months apart; and G3 (control group), no vaccination. Serological tests were carried out on Days 90, 180, 270, and 360. In each evaluation, seropositive cows were removed. The variables related to occurrence of new infections and abortion, as well as those related to subsequent reproductive performance, were analyzed using the SAS software.Results and discussionSeropositive cows were still detected in both vaccinated and control groups at 90 days. However, no new infections were detected in G1 at 180 days or in G1 and G2 at 270 and 360 days, whereas new seropositive cows were diagnosed in all exams in G3 (control). Therefore, the cumulative number of new infections was lower in vaccinated than in control cows (0.2% and 0.0%, vs. 3.2% for groups G1, G2, and G3 respectively; p = 0.0002). In farm A the number of days open was greater in the control than in vaccinated groups (p

IntroductionArtificial intelligence is revolutionizing English language teaching with personalized assessments and advanced tools, though more studies are needed on its effectiveness and equitable accessibility.MethodA systematic literature review (SLR) was conducted using articles from Web of Science, Scopus, and ERIC to establish theoretical foundations.ResultsAI in learning English as a second language offers personalized tools such as chatbots, pronunciation improvement apps, and platforms that adapt content according to student progress.DiscussionThe study highlights how AI is revolutionizing English learning by personalizing lessons, improving pronunciation, and promoting cultural understanding.

Intracytoplasmic sperm injection (ICSI) is a widely used technique in fertility centers. ICSI success depends on both nuclear and cytoplasmic oocyte maturation. Cumulus cells, which surround the oocytes, play a pivotal role in oocyte competence. However, the significance of DNA damage in cumulus cells as a marker of fertilization success remains largely unexplored. This study aims to investigate the relationship between DNA damage in cumulus cells of females undergoing ICSI, and oocyte competence, with a focus on in vitro fertilization (IVF) outcomes. We employed the alkaline comet assay to assess DNA damage levels (%TDNA) in cumulus cells and whole blood from 22 potentially fertile females and 35 infertile females, including 20 with an ovulatory disfunction phenotype. Our results revealed significant differences between the levels of %TDNA in cumulus cells and blood. Females with an ovulatory dysfunction phenotype exhibited higher levels of %TDNA in cumulus cells compared to potentially fertile females. Additionally, within the group of females with ovulatory dysfunction, a significant correlation was observed between %TDNA levels and the number of oocytes with two pronuclei. Our findings suggest that blood does not accurately reflect DNA damage in cumulus cells, which was correlated with the fertilization success in females with ovulatory dysfunction. High levels of %TDNA in cumulus cells were associated with a higher likelihood of successful fertilization. Moreover, our results imply that low levels of %TDNA may be linked to oocytes that fail to complete maturation and, consequently, do not fertilize (oocytes with zero pronuclei). Further research with larger cohorts is necessary to validate these findings and to explore potential applications in female fertility. However, our study provides evidence that DNA damage in cumulus cells may serve as a valuable biomarker for predicting fertilization success and oocyte competence.

Nanoenergetics are defined as a class of nanomaterials that possess the ability to release energy in certain situations. These properties have been studied and deepened in recent years, so much so that nanoenergetics have been introduced into the use of the weapons industry, among others. It is therefore an emerging reality that deserves attention with regard to potential harmful effects on human and environmental health. it has been suggested that nanoenergetics may have genotoxic and immunotoxic effects, among others. Problems related to exposure to nanonenergetics can therefore potentially affect both exposed workers (both in the production and use phase) and the civilian population, if used in war scenarios, for example,. Starting from these assumptions, the INNOTOX research project aims to contribute to the in-depth study of the toxicity of nanonenergetics, through an integrated approach involving experts in occupational and forensic medicine, nanotoxicology and bioengineering.

IntroductionComplement activation split products are signatures of many immunopathological disorders. Among the laboratory findings observed in these diseases, a reduction in the level of circulating intact complement components can be mentioned, and this change has also been detected in envenomation by multiple Africanized honeybee (Apis mellifera) stings. Although envenomation by these animals elicits diverse life-threatening reactions, the capacity of bee venom (AmV) to activate the human complement system remains elusive.Methods and findingsBy coupling immunochemical and functional approaches, it was observed that AmV strongly consumes components of the alternative pathway (AP) of the complement system in normal human serum (NHS). Additionally, AmV interfered with classical (CP) and lectin pathways (LP) activities. In parallel, a high increase in Ba fragment levels was detected, suggesting that the changes in AP activity were due to its activation. Furthermore, an increase in the level of the C1s-C1INH complex and a decrease in the physiological level of MASP1-C1INH suggested that CP and LP were also activated in the presence of AmV. Strikingly, NHS exposed to increasing AmV concentrations varying from 5 to 1000 µg/mL presented a high generation of C3a, C4a and C5a anaphylatoxins, and sC5b-9 complexes assembly, thus reinforcing that AmV triggers complement activation.ConclusionThese results show that AmV is a strong complement activator. This activation presents a mixed profile, with a predominance of AP activation. This suggests that complement split products can play important roles in the envenomation by Africanized honeybee, as they could induce diverse immunopathological events observed in patients and may also dictate patient clinical prognosis.

Background/aimsDue to a lack of standard pediatric prescribing information, medicines are often used in a dosage form or for an indication that has not been investigated in children. Pediatric clinical trial research networks aim to facilitate the timely availability of innovative drugs for children by developing standardized trial facilitation and conduct processes. This paper aims to assess the (pre)feasibility duration and characteristics of a US-sponsored clinical trial, in collaboration with I-ACT for Children, for distribution across European sites via European clinical research facilitation networks.MethodA transatlantic partnership between the Belgian Pediatric Clinical Research Network (BPCRN,) and I-ACT for Children conducted feasibilities in Europe for industry-sponsored early-stage pharmacological clinical trials between 2019 and 2022. The collaboration recorded time to event for key elements of feasibility, influences on successful feasibility, and benefits of collaboration.ResultsTrials were conducted across 17 European countries with 202 participating hospital sites. The initial phase, the pre-feasibility questionnaire had a 70% response rate from 142 sites, and sites took a median 38 days (IQR 20 days) to complete the questionnaire for five trials. All responses underwent a quality control, addressing inaccuracies in site capabilities and recruitment. The first trial's CDA and feasibility questionnaire were completed in roughly 2 months for 7 countries. Time to completion was affected by precontracted sites, limited scope of studies, changes in timelines, COVID-related disruptions, and a learning curve for collaboration.ConclusionCollaboration between European collaborative national networks and US-network I-ACT for Children has supported site identification of global pediatric clinical trials. This illustrates one method for the importance of early engagement with sponsors and implementation of effective communication systems.

Introduction/objectivesThe precise location of the tumor site is essential for the success of surgical treatment. Neoadjuvant chemotherapy (NAC) is a challenge for preoperative tumor and node localization. Thus, the knowledge and attitudes of the affiliated members of the Brazilian Society of Mastology (SBM) regarding breast and axilla marking were evaluated and a consensus regarding management and treatment was reached.MethodsThis was an online survey conducted between June and December 2022. All 1,742 active mastologists affiliated to the SBM were invited anonymously. The online form contained 28 objective questions, of which 22 were formulated on a Likert scale. These questions addressed relevant aspects related to breast and axilla marking in the neoadjuvant setting. Responses that reached 70% agreement were considered consensual. Statistical analysis was performed using the SPSS program version 26.0. Post hoc analysis was performed when appropriate and the significance level was set at p < 0.05. Polychoric regression analyses were conducted using `VGAM` packageResultsIn total, 468 mastologists answered the questionnaire (26.8%), with a predominance of professionals aged between 40–49 years (32.1%). Most professionals were board-certified (84,8%). The indication of tumor marking in the breast prior to NAC was consensual (96.4%) and the metal clip was the preferred method (69.7%). There was no consensus regarding the indication of pre-NAC histologically positive lymph node marking (49.8% disagree and 42.8% agree). However, there was consensus that the clinical and imaging evaluation was insufficient for staging the axilla as N1 (71.6%). The contraindication of breast and node marking in T4b tumors (71.2%) was consensual. There was consensus on the indication of sentinel lymph node biopsy (SLNB) for initially cN1 (92.3%) or cN2 (72.7%) tumors that became cN0 after NAC, with 67.5% opting for dual staining with technetium and patent blue. When

The rise of antibiotic-resistant bacterial strains has become a critical health concern. According to the World Health Organization, the market introduction of new antibiotics is alarmingly sparse, underscoring the need for novel therapeutic targets. The LytR-CpsA-Psr (LCP) family of proteins, which facilitate the insertion of cell wall glycopolymers (CWGPs) like teichoic acids into peptidoglycan, has emerged as a promising target for antibiotic development. LCP proteins are crucial in bacterial adhesion and biofilm formation, making them attractive for disrupting these processes. This study investigated the structural and functional characteristics of the LCP domain of LytR from Streptococcus dysgalactiae subsp. dysgalactiae. The protein structure was solved by X-ray Crystallography at 2.80 Å resolution. Small-angle X-ray scattering (SAXS) data were collected to examine potential conformational differences between the free and ligand-bound forms of the LytR LCP domain. Additionally, docking and molecular dynamics (MD) simulations were used to predict the interactions and conversion of ATP to ADP and AMP. Experimental validation of these predictions was performed using malachite green activity assays. The determined structure of the LCP domain revealed a fold highly similar to those of homologous proteins while SAXS data indicated potential conformational differences between the ligand-free and ligand-bound forms, suggesting a more compact conformation during catalysis, upon ligand binding. Docking and MD simulations predicted that the LytR LCP domain could interact with ADP and ATP and catalyze their conversion to AMP. These predictions were experimentally validated by malachite green activity assays, confirming the protein’s functional versatility. The study provides significant insights into the structural features and functional capabilities of the LCP domain of LytR from S. dysgalactiae subsp. dysgalactiae. These findings pave the way for designing targeted therapies against antibiotic-resistant bacteria and offer strategies to disrupt bacterial biofilm formation.

Integrated systems are efficient technologies used for diversifying production and promoting agricultural sustainability. The benefits of double intercropping are well-established in research. With advancements in sustainable food production technology, some studies have explored triple intercropping by adding legumes. However, triple intercropping methods must be comprehensively understood. This study aimed to evaluate the production of dry mass, fermentation profile, and nutritive values of silage from maize based intercropping. The experiment was conducted in the field, in Instituto Federal Goiano, Campus Rio Verde, State of Goiás, Brazil, where we assessed silage production and quality in monoculture and intercropping systems, in a randomized block complete design with three replications. We evaluated 8 treatments, consisting of silage from: maize monoculture; Tamani guinea grass monoculture; Quênia guinea grass monoculture; Zuri guinea grass monoculture; Pigeon pea monoculture; maize + Tamani guinea grass + Pigeon pea; maize + Quênia guinea grass + Pigeon pea and maize + Zuri guinea grass + Pigeon pea. The results showed that maize silage intercropped with cultivars of Panicum maximum (Tamani, Quênia and Zuri guinea grasses) and Pigeon pea increased silage mass production by 37.8% and the crude protein content at 25.1% compared with that of the monoculture maize silage. It also improved the fermentation characteristics of silage, reducing 5.1% of pH, 32.5% of buffering capacity, 28.9% of ammoniacal nitrogen in total nitrogen, 15.7% of effluent production, and 20.0% of dry matter losses, compared with those of grass and legume silage in monoculture. Thus, the silage produced by the triple intercropping proved to be an efficient technique for sustainable forage production.

BackgroundPeople with structural vulnerabilities (including immigrants, people who use drugs, and those who are unhoused or uninsured) are more likely to experience COVID-19 testing disparities relative to other groups. We documented barriers and facilitators to COVID-19 testing and explored how structural vulnerabilities created and/or exacerbated COVID-19 testing barriers.MethodsBetween 2021 and 2022, we conducted semi-structured interviews with 34 members of structurally vulnerable populations and 27 key informants who provide health and social services to them. Our abductive analysis was iterative, utilizing both inductive and deductive coding processes. Recognizing that adequate and appropriate testing for COVID-19 is a complex health behavior that involves both decision-making and issues related to access, we developed a hybrid model of COVID-19 testing behavior to organize reported barriers. We then used that model for more in-depth analysis of structural vulnerabilities in the context of testing.ResultsOur model of testing behaviors provides a framework for understanding the many barriers and facilitators relevant to COVID-19 testing. After identifying locally-reported barriers, we found that specific conditions—economic precarity, legal precarity, the confusing U.S. healthcare landscape, English-exclusive environments, and stigmatizing medical encounters—make adequate and appropriate testing less likely by making COVID-19 testing feel riskier (entailing legal, financial, and psycho-social risks) and making healthcare, and thus vicariously testing, more difficult to access.ConclusionThe COVID-19 pandemic exposed disparities in health care delivery. To avoid under-testing and its associated health consequences during the next pandemic, public health efforts should address structural conditions to ameliorate risks and bolster testing infrastructure to improve access.

Tuberculosis remains a serious global health threat, exacerbated by the rise of resistant strains. This study investigates the potential of two benzofuroxan (Bfx) derivatives, 5n and 5b, as targeted treatments for MDR-TB using in silico, in vitro, and in vivo methodologies. In vitro analyses showed that Bfx compounds have significant activity against Mtb H37Rv, with Bfx 5n standing out with a MIC90 of 0.09 ± 0.04 μM. Additionally, their efficacy against MDR and pre-XDR strains was superior compared to commercial drugs. These Bfx compounds have a narrow spectrum for mycobacteria, which helps avoid dysbiosis of the gut microbiota, and they also exhibit high selectivity and low toxicity. Synergism studies indicate that Bfx derivatives could be combined with rifampicin to enhance treatment efficacy and reduce its duration. Scanning electron microscopy revealed severe damage to the morphology of Mtb following treatment with Bfx 5n, showing significant distortions in the bacillary structures. Whole-genome sequencing of the 5n-resistant isolate suggests resistance mechanisms mediated by the Rv1855c gene, supported by in silico studies. In vivo studies showed that the 5n compound reduced the pulmonary load by 3.0 log10 CFU/mL, demonstrating superiority over rifampicin, which achieved a reduction of 1.23 log10 CFU/mL. In conclusion, Bfx derivatives, especially 5n, effectively address resistant infections caused by Mtb, suggesting they could be a solid foundation for future therapeutic developments against MDR-TB.

IntroductionCongenital hypopituitarism (CH) is characterized by the deficiency of pituitary hormones. Among CH patients, 85% lack a molecular diagnosis. Whole Exome Sequencing (WES) identified a homozygous variant (c.865G>A, p.Val289Ile) in the CDH2 gene, responsible for N-Cadherin production, crucial for cell-cell adhesion. Predicted to be likely pathogenic, the variant was found in a patient deficient in GH, TSH, ACTH, and LH/FSH. Its impact on cell adhesion was confirmed in L1 fibroblast cell lines.ObjectiveCreate a cdh2 knockdown in zebrafish for investigating its role in pituitary development through growth hormone and transcription factors expression.MethodsUtilized pET28B-RfxCas13d-His plasmid for Cas13 mRNA production via in vitro transcription, guiding Cas13 to cdh2 with three RNAs. Injected the complex into single-cell embryos for analysis up to 96 hpf. Assessed gene expression of cdh2, prop1, pit1, and gh1 using RT-qPCR. Evaluated cdh2 protein expression through the western blot technique.ResultsKnockdown animals displayed developmental delay. The cdh2 expression decreased by 75% within 24 hours, rebounded by 48 hours, and reached wild-type levels by 96 hpf. gh1 expression decreased at 48h but increased by 96 hpf, aligning with WT. No significant differences in prop1 and pit1 expression were observed.ConclusionOur findings underscore cdh2’s role in pituitary development and hormonal regulation, offering insights for developmental biology research.

IntroductionColorectal cancer (CRC) is the third most incident and the second most lethal malignant tumor. Despite the recognized association between obesity and CRC, further clarification is necessary regarding the lipids that are overexpressed during the development of CRC. In this scenario, the combination of metabolomics and a three-dimensional (3D) co-culture model involving CRC tumor cells and lipids can enhance the knowledge of energy metabolism modifications at the cross-talk between colorectal cancer and adipocytes. This study aimed to screen potential metabolites in the three dimensional (3D) co-culture of CRC and adipocytes by investigating the metabolome composition of this co-culture released into the extracellular space, which is known as the secretome.MethodsPre-adipocyte cells (3T3-L1), human colon carcinoma (HT-29), and the 3D co-culture (3T3-L1 + HT-29) were cultured for the secretome obtention. Then, ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) was employed to analyze the metabolomics of each secretome.ResultsOverall, 3.731 molecules were detected independent of the cell culture. When comparing the three cultures, 105 molecules presented a statistically significant difference in abundance between groups. Among these molecules, 16 were identified, with a particular emphasis on six lipids (PG 20:0, octadecenal, 3-Hydroxytetracosanoyl-CoA, 9,10-dihydroxy-octadecenoic acid, palmitoleic acid, and PA 18:4) and one amino acid derivative (acetylglutamic acid), which presented significant scores during the partial least-squares discriminant analysis (PLS-DA).DiscussionAlthough it is too early to determine the possible impact of such molecules in a CRC microenvironment, these results open new avenues for further studies on the energy metabolism at the cross-talk of colorectal cancer adipocytes.

IntroductionThe development of reading and complex executive functions is fundamental for achieving social, academic, and professional success. So far, there is no single neuropsychological instrument that comprehensively assesses the domains of inhibitory control, cognitive flexibility, working memory, and reading comprehension. To assess executive functions related to reading, the “Assessment of Reading and Executive Functions” (AREF) was developed. In this study, we show initial evidence of validity and reliability for four subtests - Graphophonological-Semantic Flexibility, Inhibitory Control, Flexibility, and Working Memory.MethodsA total of 93 students from 4th to 9th grade, aged 8-14, in public (n = 61) and private (n = 32) schools were evaluated. Tasks from the AREF instrument, as well as measures of reading comprehension, inhibitory control, cognitive flexibility, working memory, and intelligence, were administered. Correlations between AREF scores and the other measures were performed to assess external construct validity. Performance differences between school groups on AREF subtests were analyzed using ANOVA, t-test, and Mann-Whitney tests, and the internal consistency of the instrument’s tasks was evaluated using Cronbach’s alpha coefficient.ResultsThe scores of the AREF subtests demonstrated significant positive correlations with reading measures (ranging from 0.339 to 0.367) and executive functions (ranging from 0.209 to 0.396). Significant differences were found in the performance of some AREF tasks when comparing individuals from public and private schools, as well as between 4th and 5th graders compared to students in higher grades. The internal consistency of the tasks was low for Graphophonological-Semantic Flexibility (Cronbach’s α = 0.566), moderate for Inhibitory Control and Flexibility (Cronbach’s α = 0.768), and high for Working Memory (Cronbach’s α = 0.881).DiscussionThe results provide initial evidence of construct validity and reliability for the AREF subtests. It is expected that this new neuropsychological test will contribute to the assessment of reading skills and executive functions, assisting in guiding clinical and educational interventions for individuals with and without neurodevelopmental disorders.

AimsThe integration of smartwatches into postoperative cardiac care transforms patient monitoring, systematically tracking vital signs and delivering real-time data to a centralized platform. This study focuses on developing a platform for seamless integration, assessing reliability, and evaluating the impact on post-cardiac surgery. The goal is to establish a robust foundation for understanding the efficacy and dependability of smartwatch-based telemonitoring, enhancing care for this population.Methods and resultsA total of 108 cardiac surgery patients were divided into telemonitoring (TLM) and control (CTL) groups. The TLM group utilized smartwatches for continuous monitoring of vital parameters (SpO2, HR, BP, ECG) over 30 ± 3 days. Statistical analyses (Pearson, Intraclass Correlation, Bland-Altman, Tost Test) were employed to compare smartwatch measurements with traditional methods. Significant correlations and concordance were observed, particularly in HR and BP measurements. Challenges were noted in SpO2 measurement. The ECG algorithm exhibited substantial agreement with cardiologists (Kappa: 0.794; p > 0.001), highlighting its reliability. The telemonitoring platform played a crucial role in early detection of clinical changes, including prompt Emergency Department (ED) visits, contributing significantly to preventing outcomes that could lead to mortality, such as asymptomatic Atrioventricular block. Positive patient responses affirmed technological efficacy, especially in identifying cardiac arrhythmias like atrial fibrillation.ConclusionThe integration of smartwatches into remote telemonitoring for postoperative cardiac care demonstrates substantial potential, improving monitoring and early complication detection, thereby enhancing patient outcomes. The FAPO-X Study (Assisted Digital Telemonitoring with Wearables in Patients After Cardiovascular Surgery; NCT05966857) underscores the promising role of telemonitoring in postoperative cardiac care.

Clinical relevanceThis study compares a novel photoscreening device with a previously validated one in a school-age population. It highlights a tendency of the new device to underestimate myopic spherical equivalent and overestimate hyperopic cases.PurposeTo compare the PlusoptiX A16 and Vision Screener V100 photoscreeners in a study population of school-age children.MethodsOne hundred and thirty-three children, with a mean age of 6.4 ± 0.5 years, were evaluated using both the PlusoptiX A16 and Vision Screener V100 photoscreeners. The measurements were taken in random order in a room with diminished ambient lighting.ResultsThe mean refractive error values for the M component were 0.27 ± 0.67D (PlusoptiX A16) and 0.21 ± 0.58D (Vision Screener V100). For the J0 component, means were 0.16 ± 0.38D (PlusoptiX A16) and 0.06 ± 0.33D (Vision Screener V100) and for theJ45 component the means were 0.03 ± 0.17D (PlusoptiX A16) and 0.06 ± 0.22D (Vision Screener V100). When compared both instruments, statistically significant differences were observed for the M (p=0.017) and J0 (p=0.004) components. The agreement rates between PlusoptiX A16 and Vision Screener V100 across different refractive components were 80.5% for sphere, 82.0% for cylinder, and 40.6% for axis when considering a range of ±0.75 D for sphere and cylinder and ±25.0 degrees for cylinder axis. Simultaneously considering all three conditions, the overall agreement was 73.7%.ConclusionThe Vision Screener V100, while generally aligning well with PlusoptiX A16, tends to underestimate myopic spherical equivalent, overestimate hyperopic cases, and underestimate J0 astigmatism.

The formation of a lytic immunological synapse (IS) is crucial for cytotoxic lymphocytes to accurately target and effectively eliminate malignant cells. While significant attention has been focused on the lymphocyte side of the IS, particularly its role as a secretory domain for lytic granules, the cancer cell side of the IS has remained relatively underexplored. Recent findings have revealed that cancer cells can rapidly polarize their actin cytoskeleton toward the IS upon interaction with natural killer (NK) cells, thereby evading NK cell-mediated cytotoxicity. In this Brief Research Report, we present preliminary findings suggesting that actin cytoskeleton remodeling at the cancer cell side of the IS is associated with the targeted secretion of small extracellular vesicles towards the interacting NK cell. We observed that multivesicular bodies (MVBs) preferentially accumulate in the synaptic region in cancer cells exhibiting synaptic accumulation of F-actin, compared to those lacking actin cytoskeleton remodeling. Extracellular immunofluorescence staining revealed increased surface exposure of CD63 at the cancer cell side of the IS, suggestive of the fusion of MVBs with the plasma membrane. This hypothesis was supported by a pH-sensitive probe demonstrating dynamic trafficking of CD63 to the extracellular region of the IS. Collectively, our data support the notion that cancer cells can engage in targeted secretion of extracellular vesicles in response to NK cell attack, underscoring the need for further research into the potential role of this process in facilitating cancer cell immune evasion.

BackgroundDespite the growing number of elderly kidney transplant (Ktx) recipients, few studies have examined the effects of immunosuppression on their lymphocyte profiles.MethodsWe evaluated the early conversion from mycophenolate sodium (MPS) to everolimus (EVL) after rabbit antithymocyte globulin (rATG) 2 mg/kg induction in elderly kidney recipients. Three groups of KTx patients were compared: (a) Young (n=20, 36 ± 7 y) receiving standard immunosuppression (Group A1) (prednisone, tacrolimus, and MPS), (b) Elderly (n=35, 65 ± 3 y) receiving standard immunosuppression (Group B1), and (c) Elderly (n=16, 65 ± 3 y) with early (mean 30 d) conversion from MPS to EVL (Group B2). Naive, memory, and regulatory peripheral blood TCD4+ lymphocytes were quantified at 0, 30, and 365 d.ResultsResults are reported as [mean(p25–p75)]. Young recipients had higher lymphocyte counts at baseline [2,100(1,630–2,400) vs. 1,310 (1,000–1,600)/mm3, p

IntroductionIn adult horses, the development and evaluation of enteral electrolyte solutions containing sodium acetate for correcting hyperchloremic metabolic acidosis are still lacking, although these electrolyte and acid–base imbalances are commonly observed. The objective of this study was to evaluate the alkalinizing effect of two enteral electrolyte solutions containing different concentrations of acetate, administered via nasogastric tube in continuous flow, in adult horses with experimental hyperchloremic metabolic acidosis.MethodsSix mares aged between 3 and 10 years were used in a 6×2 crossover design, with each animal receiving both treatments. The horses were subjected to a protocol to induce hyperchloremic metabolic acidosis. They then received one of two treatments: HighAcetate (81.4 mmol/L) and LowAcetate (22.7 mmol/L) at an infusion rate of 15 mL/kg/h for 12 h. Plasma, serum, and urinary biochemical assessments; hematocrit; urinary volume, pH, and specific gravity; and blood gas analysis were measured at the following time points: T-12 h (beginning of the 12-h fast), T0h (end of fasting and beginning of the acidosis induction phase), every 2 h during the hyperchloremic metabolic acidosis induction phase (Ti2h and Ti4h), every 2 h during the 12-h enteral hydration phase (Tt2h, Tt4h, Tt6h, Tt8h, Tt10h, and Tt12h), with one sample taken at T24h (24 h after the start of acidosis induction) and another at T36h (36 h after the start of acidosis induction). Data were analyzed using descriptive statistics and analysis of variance based on a factorial design of repeated measures, with Tukey’s post-hoc test or the Kruskal-Wallis test with Dunn’s post-hoc test for non-parametric tests.ResultsAt the end of the induction phase, the animals developed moderate to severe hyperchloremic metabolic acidosis. The HighAcetate solution effectively corrected electrolyte and acid–base imbalances before the end of the treatment phase (Tt12h), while the LowAcetate solution was not effective in correcting those changes.ConclusionThe HighAcetate (81.4 mmol/L) solution is deemed an effective and safe alternative for the treatment of hyperchloremic metabolic acidosis in horses.

Over the last few decades, advancements in process safety and quality methods have been significantly improved, yet new challenges continue to emerge in the sustainable food supply chain. This study aimed to investigate some physicochemical and microbiological parameters impacting meat products, particularly cooked sausages, within a sustainable supply chain, focusing on quality, spoilage populations, and syneresis formation under vacuum conditions. A comprehensive analysis was conducted on 355 samples collected over four seasons using high-throughput sequencing (16S/ITS) and microbiological and physicochemical [pH and water activity (aw)] assessments. The microbial growth predictor MicroLab_ShelfLife was employed, and multiple factor analysis (MFA) and agglomerative hierarchical clustering (AHC) were utilized to understand how these variables influence the microbiome resilience of these products. Lactic and acetic acids were correlated with the microbiome of the sausages and the liquid coating covering them using metagenomic analyses. The study highlighted that 52% of the evaluated meat industries in southeastern Brazil are implementing effective protocols for sustainable chain production. The results indicated that the durability of vacuum-packaged cooked sausages was primarily influenced by storage temperature (RV coefficient of 0.906), initial microbial load (0.755), and aw (0.624). Average microbial counts were 4.30 log cfu/g (initial), 4.61 (7°C/4 days), 4.90 (7°C/8 days), 6.06 (36°C/4 days), and 6.79 (36°C/8 days). Seasonal durability analysis revealed that winter had the highest average durability of 45.58 days, while summer had the lowest at 26.33 days. Yeast populations, including Trichosporon sp. and Candida sp., were identified as key genera influencing spoilage dynamics. In addition, Bacillus species emerged as dominant spoilage microorganisms, highlighting the need for new critical controls. This study demonstrates the impact of metagenomic approaches, including ITS and 16S amplicon sequencing, in revealing microbial community dynamics, storage temperature, and aw, which are essential for developing targeted interventions to enhance food safety and quality sustainably.

Despite treatment advances through immunotherapies, including anti-PD-1/PD-L1 therapies, the overall prognosis of non-small cell lung cancer (NSCLC) patients remains poor, underscoring the need for novel approaches that offer long-term clinical benefit. This review examined the literature on the subject over the past 20 years to provide an update on the evolving landscape of dendritic cell-based immunotherapy to treat NSCLC, highlighting the crucial role of dendritic cells (DCs) in immune response initiation and regulation. These cells encompass heterogeneous subsets like cDC1s, cDC2s, and pDCs, capable of shaping antigen presentation and influencing T cell activation through the balance between the Th1, Th2, and Th17 profiles and the activation of regulatory T lymphocytes (Treg). The intricate interaction between DC subsets and the high density of intratumoral mature DCs shapes tumor-specific immune responses and impacts therapeutic outcomes. DC-based immunotherapy shows promise in overcoming immune resistance in NSCLC treatment. This article review provides an update on key clinical trial results, forming the basis for future studies to characterize the role of different types of DCs in situ and in combination with different therapies, including DC vaccines.

IntroductionOverusing and misusing pesticides, including paraquat (PQ), have led to numerous environmental contamination complications. PQ is an emerging bio-accumulative contaminant that is present in environmental aqueous matrices. Adsorption techniques are part of a set of technologies applied in ecological remediation, known for their high effectiveness in removing aqueous PQ. A study of the PQ adsorption capacity of three cyclodextrin–silica nanocomposites (α-CDSi, β-CDSi, and γ-CDSi) from contaminated waters is presented in this paper.MethodsThe cyclodextrin–silica nanocomposites were synthesized via an esterification reaction between the inorganic matrix and cyclodextrins (CDs) (α, β, and γ) and were characterized physicochemically by spectroscopic, thermal, and surface methods. Their PQ removal performance from contaminated aqueous media was studied under different experimental conditions.Results and DiscussionThe results showed a fast adsorptive response in removal treatment studies over time. Adsorption capacities of 87.22, 57.17, and 77.27 mg.g−1 were found for α-CDSi, β-CDSi, and γ-CDSi, respectively, at only 30 min of treatment. Thermodynamic studies indicated spontaneous and exothermic adsorption processes. The removal assays responded mainly to physisorption mechanisms with contributions from chemisorption mechanisms. Spectroscopic assays showed a strong interaction of PQ with the adsorbents used. Innovative CDSi nanocomposites have proven to be highly efficient in applying aqueous PQ remediation, thus proving to be sustainable adsorbents of contaminants of emerging importance worldwide.

ObjectivesTo establish a consensus for evaluating second victims (SV) support interventions to facilitate comparison over time and across different organizations.MethodsA three-phase qualitative study was conducted from June 2023 to March 2024. This consensus approach engaged members of the European Researchers Network Working on Second Victims. A nominal group technique and insights from a scoping review were used to create a questionnaire for Delphi Rounds. Indicators were rated 1–5, aiming for agreement if over 70% of participants rated an indicator as feasible and sensitive with scores above 4, followed by a consensus conference.ResultsFrom an initial set of 113 indicators, 59 were assessed online, with 35 advancing to the Delphi rounds. Two Delphi rounds were conducted, achieving response rates of over 60% and 80% respectively, resulting in consensus on 11 indicators for evaluating SV support programs. These indicators encompass awareness and activation, outcomes of SV support programs, as well as training offered by the institution.ConclusionThis study presents a scoreboard for designing and monitoring SV support programs, as well as measuring standardized outcomes in future research.

ObjectivesThis study was performed to identify predictive markers of worse outcomes in patients with severe COVID-19 in an intensive care unit.MethodsSixty patients with severe COVID-19, hospitalized in the Intensive Care Unit (ICU) between March and July 2021, were stratified into two groups according to the outcome survivors and non-survivors. After admission to the ICU, blood samples were collected directly for biomarker analysis. Routine hematological and biochemical biomarkers, as well as serum levels of cytokines, chemokines, and immunoglobulins, were investigated.ResultsLymphopenia, neutrophilia, and thrombocytopenia were more pronounced in non-surviving patients, while the levels of CRP, AST, creatinine, ferritin, AST, troponin I, urea, magnesium, and potassium were higher in the non-surviving group than the survival group. In addition, serum levels of IL-10, CCL2, CXCL9, and CXCL10 were significantly increased in patients who did not survive. These changes in the biomarkers evaluated were associated with increased mortality in patients with severe COVID-19.ConclusionThe present study confirmed and expanded the validity of laboratory biomarkers as indicators of mortality in severe COVID-19.

PurposeThis study aimed to determine the prevalence of anxiety and depression among patients with glaucoma compared to the average Brazilian prevalence (9.8% of anxiety and 5.8% of depression, according to the World Health Organization) and its correlation with the severity of the disease.MethodsThis was a transversal, single-arm trial of patients from four glaucoma centers in São Paulo and Curitiba—Brazil. Patients comprised adults at least 18 years of age with glaucoma diagnosis under treatment for at least 6 months. All subjects of the study answered two questionnaires (PHQ-9 and GAD-7) to evaluate the presence of anxiety and depression, and the results were analyzed accordingly to clinical and demographic characteristics.ResultsThe protocol included a total of 210 patients. The average age was 61.6 ± 15.3 years, and the female gender was more common (68.86%). Primary open-angle glaucoma was the most common diagnosis (59.90%). The average IOP was 18.5 mmHg, and 1.5 anti-glaucoma drops were the mean treatment. The prevalence of depression and anxiety was 26.90 and 25.71%, respectively. Most patients with anxiety were classified as early glaucoma, while those with depression had severe glaucoma.ConclusionThis study found that the prevalence of anxiety and depression among patients with glaucoma is higher than in the general population in our country.

Bone substitutes are widely employed for applications in orthopedic surgery for the replacement of injured bone. Among the diverse methods that are used to design 3D bioceramic matrices, Freeze Foaming has gained attention, since it provides the ability to tune the shape of the created structures. One of the major problems related to these constructs is the lack of porosity at the outwards sides (holder) of the scaffold, thus reducing the cellular affinity and creating a rejection of the implant. In this research, we aimed to develop a bone scaffold with enhanced surface properties and improved cellular affinity. The main aim was to alter the biocompatibility characteristics of the 3D bioceramic constructs. We have produced three-dimensional, complex-shaped hollow shell structures, manufactured by Additive Manufacturing processes and as a second step, filled with a ceramic suspension by the Freeze-Foaming process. 3D constructs from HAP-derived TCP and TCP/ZrO2 were synthesized by freeze-foaming method and subsequently irradiated with a fs-laser (λ = 800 nm) spanning a range of parameters for achievement of optimal surface processing conditions. The designed scaffolds demonstrated enhanced topographical properties with improved porosity examined by SEM, EDX, and 3D profilometry after laser treatment. Wettability and computer tomography (CT) evaluation was also performed. The results from X-ray diffraction (XRD) and micro-Raman analysis did not show photochemical and surface or volume defects and changes after laser processing of the ceramic samples. Preliminary results from MG-63 osteoblast-like cell tests showed good cell affinity on the processed surfaces and no cytotoxic effect on the cells.

IntroductionVisceral leishmaniasis (VL) is an important tropical and neglected disease and represents a serious global health problem. The initial interaction between the phagocytes and the parasite is crucial to determine the pathogen’s capacity to initiate infection and it shapes the subsequent immune response that will develop. While type-1 T-cells induce IL-6, IL-1β, TNF-α, and IL-12 production by monocytes/macrophages to fight the infection, type-2 T-cells are associated with a regulatory phenotype (IL-10 and TGF-β) and successful infection establishment. Recently, our group demonstrated the role of an important Th1/Th17 T-cell population, the mucosal-associated invariant T (MAIT) cells, in VL. MAIT cells can respond to L. infantum by producing TNF-α and IFN-γ upon MR1-dependent activation.Objective and methodsHere, we describe the impact of the MR1-blockage on L. infantum internalization on the functional profile of circulating neutrophils and monocytes as well as the impact of the MR1-blockage on the soluble mediator signatures of in vitro whole blood cultures.ResultsOverall, our data showed that VL patients presents higher percentage of activated neutrophils than asymptomatic and non-infected controls. In addition, MR1 blockade led to lower TNF-α and TGF-β production by non-activated neutrophils from asymptomatic individuals. Moreover, TNF-α and IL-10 production by monocytes was higher in VL patients. In the analysis of soluble mediators produced in vitro, MR1-blockade induced a decrease of IFN-γ and an increase of IL-10, IL-27 and IL-33 in the cell cultures of AS group, a cytokine pattern associated with type 2 deleterious response.Discussion and conclusionThese data corroborate the hypothesis that MR1-restricted responses are associated to a protective role during Leishmania infection.

Phytophthora cinnamomi Rands devastates forest species worldwide, causing significant ecological and economic impacts. The European chestnut (Castanea sativa) is susceptible to this hemibiotrophic oomycete, whereas the Asian chestnuts (Castanea crenata and Castanea mollissima) are resistant and have been successfully used as resistance donors in breeding programs. The molecular mechanisms underlying the different disease outcomes among chestnut species are a key foundation for developing science-based control strategies. However, these are still poorly understood. Dual RNA sequencing was performed in C. sativa and C. crenata roots inoculated with P. cinnamomi. The studied time points represent the pathogen’s hemibiotrophic lifestyle previously described at the cellular level. Phytophthora cinnamomi expressed several genes related to pathogenicity in both chestnut species, such as cell wall–degrading enzymes, host nutrient uptake transporters, and effectors. However, the expression of effectors related to the modulation of host programmed cell death (elicitins and NLPs) and sporulation-related genes was higher in the susceptible chestnut. After pathogen inoculation, 1,556 and 488 genes were differentially expressed by C. crenata and C. sativa, respectively. The most significant transcriptional changes occur at 2 h after inoculation (hai) in C. sativa and 48 hai in C. crenata. Nevertheless, C. crenata induced more defense-related genes, indicating that the resistant response to P. cinnamomi is controlled by multiple loci, including several pattern recognition receptors, genes involved in the phenylpropanoid, salicylic acid and ethylene/jasmonic acid pathways, and antifungal genes. Importantly, these results validate previously observed cellular responses for C. crenata. Collectively, this study provides a comprehensive time-resolved description of the chestnut–P. cinnamomi dynamic, revealing new insights into susceptible and resistant host responses and important pathogen strategies involved in disease development.

In this study, we explored the effects of chronic stress on lumpfish (Cyclopterus lumpus) physiological, immune response, health, and plasma free amino acids. 3 groups of lumpfish were exposed to 1-minute air exposure. 1 group was exposed to stress once per week, a second group exposed 2 times per week, and a third group exposed 4 times per week. The present study revealed significant alterations in immunity and increased nutritional demands, particularly the branched chain amino acids and lysine. Cortisol levels fluctuated, with significantly higher levels halfway through the experiment on the groups that were stressed more often. Though, by the end of the experiment, there were no significant differences in cortisol levels between groups. Regardless of stress exposure, cataract developed in virtually all sampled fish, pointing toward a potential dietary imbalance. A transient immunomodulation of stress was visible. While in early stages stress had an immune enhancing effect, as seen by the increase in plasma nitric oxide and peroxidase in the group most frequently exposed to stress, these differences were not apparent by the end of the experiment. Additionally, the worst health condition was found in this group. Our results underscore the complex interplay between stress, immunity and nutrition, highlighting the need for tailored dietary strategies and improved rearing practices.

BackgroundIn osteoarthritis (OA), articular homeostasis is regulated by microRNA-140 that inhibits ADAMTS-5, an enzyme that cleaves aggrecan and stimulates the synthesis of other inflammatory mediators. This study aims to evaluate the expression of microRNA-140 in extracellular vesicles (EVs) derived from equine synovial-membrane-derived mesenchymal stem cells (eqSMMSCs) cultured in monolayer (2D) and three-dimensional (3D) culture models under an in vitro inflammatory environment.MethodsFour experimental groups of eqSMMSC cultures were defined for isolation of the EVs. The 2D and 3D control groups were cultured in a conventional cell culture medium, while the 2D-OA and 3D-OA treatment groups were exposed to an OA-like medium containing IL-1β and TNFα. The culture media samples were collected at 24 h, 72 h, and 120 h time points for EV isolation and characterization using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Reverse transcription quantitative polymerase chain reaction was employed to assess the expressions of microRNA-140 in both the cells and EVs. All statistical analyses were conducted at the 5% significance level.ResultsEncapsulation of the eqSMMSCs protected the cells from the inflammatory media compared to the monolayer cultures. EVs were found in higher concentrations in the 3D-OA cultures. Additionally, higher expressions of microRNA-140 were observed in the cells of the 3D-OA group at 24 and 72 h, whereas microRNA-140 expressions in the EVs were higher in the 3D group at 72 h and in the 2D-OA group at 120 h (p < 0.001). However, the 3D-OA culture showed higher expression of the mRNA Adamts5 in the EVs at 120 h.ConclusionThe responses of the eqSMMSCs to inflammatory stimuli involve intracellular expression of microRNA-140 and its subsequent transportation via the EVs, with quicker responses observed in the 3D than 2D cultures. This study sheds light on the behaviors of stem cells in restoring homeostasis in osteoarthritic joints.

Satellite detection of active fires has contributed to advance our understanding of fire ecology, fire and climate dynamics, fire emissions, and how to better manage the use of fires as a tool. In this study, we use active fire data of 12 years (2012–2023) combined with landcover information in the South-Central United States to derive a monthly, open-access dataset of categorized fires. This is done by calculating a fire predominance index used to rank fire-prone landcovers, which are then grouped into four main landscapes: grassland, forest, wildland, and crop fires. County-level aggregated analyses reveal spatial distributions, climatologies, and peak fire months that are particular to each fire type. Using the Standardized Precipitation Index (SPI), it was found that during the climatological fire peak-month, the SPI and fires exhibit an inverse relationship in forests and crops, whereas grassland and wildland fires show less consistent inverse or even direct relationship with the SPI. This varied behavior is discussed in the context of landscapes’ responses to anomalies in precipitation and fire management practices, such as prescribed fires and crop residue burning. In a case study of Osage County (OK), we find that large wildfires, known to be closely related to climate anomalies, occur where forest fires are located in the county and absent in areas of grassland fires. Weaker grassland fire response to precipitation anomalies can be attributed to the use of prescribed burning, which is normally planned under environmental conditions that facilitate control and thus avoided during droughts. Crop fires, on the other hand, are set to efficiently burn residue and are practiced more intensely in drier years than in wetter years, explaining the consistently strong inverse correlation between fires and precipitation anomalies. In our increasingly volatile climate, understanding how fires, vegetation, and precipitation interact has become imperative to prevent hazardous fire conflagrations and to better manage ecosystems.

BackgroundA better understanding of the consequences of the Coronavirus Disease 2019 (COVID-19) pandemic on lifestyle of patients with Chagas disease (ChD) is of paramount importance to facilitate the implementation of intervention strategies tailored to this specific population.ObjectiveThe present study aimed to evaluate the level of physical activity (PA) in Chagas disease (ChD) patients during the Coronavirus Disease 2019 (COVID-19) pandemic and its main associated factors.MethodsThis is a cross-sectional study with 187 patients of both sexes, aged ≥18 years, followed in a national infectious disease center (Rio de Janeiro, Brazil). The level of PA was determined by the International Physical Activity Questionnaire short version and expressed in terms of total volume of physical activity (PA) (MET-minutes per week). Individuals were classified as physically active following the 2020 World Health Organization PA guideline. The exposure variables were age, sex, race, marital status, schooling, income per capita, number of rooms per domicile, number of residents per domicile, body mass index, clinical form of ChD, COVID-19 antibodies, comorbidities, self-reported anxiety, self-reported depression, self-reported fear, and self-reported sadness. The association between the exposure variables with total PA (as a continuous variable) was determined using univariate and multivariate linear regression models.ResultsMean age was 61.1 ± 11.6 years. Most (62%) were women and self-declared their race as mixed (50.8%). The percentage of physically active individuals according to was 52%. The variables independently associated with total PA levels were non-white race (Exp β = 1.39; 95% CI 1.02 to 1.90), dyslipidemia (Exp β = 0.73; 95% CI 0.56 to 0.95) and self-reported depression during quarantine (Exp β = 0.71; 95% CI 0.52 to 0.96).ConclusionNon-white race was positively associated with total levels of PA, while dyslipidemia, and self-reported depression during quarantine were negatively associated with total levels of PA. The identification of associated factors can facilitate the development of tailored strategies to increase PA levels ChD patients.

Physical inactivity can lead to frailty and negative health outcomes in middle-aged to older adults. Sedentary individuals have double the risk of death compared to those who engage in high levels of physical activity (PA). The advantages of practicing PA in older age are significant, with regular, moderate-intensity activity (150 min per week)being consistently linked with a decreased risk of chronic disease, cognitive decline, and mortality. Therefore, the study aimed to carry out a bibliometric review related to the terms “Physical activity,” “Functional capacity” and “Aging” including all the documents published in the Web of Science Core Collection until 31st December 2023. The sample was made up of 231 studies related to the topic. The results reported that the first document was published in 1994. However, there was no continuity in the publication of the documents till 1998, which was the first year with at least one document published. Considering 1998 as the first year, it is observed an exponential growth of 77.4%, between the oldest (1997–2008) and contemporaneous studies (2008–2023), in which “Geriatric Gerontology” was the Web of Science category with the highest number of documents (n = 59). The journal “Experimental Gerontology” was associated with the largest number of published documents (n = 7), being indexed in Quartil 2. The years 2009 had the highest number of citations (n = 1811), with a total of 7 documents published and 2018 with the higher number of documents (n = 25). These results reported the importance of PA in elderly people, and how it influences the risk of falls, improving the balance and the functional capacity. Thus, it is important to carry out programmes to promote physical activity to this population and reduce the risk of falls and the presence of diseases.

Age-related neurodegenerative diseases, like Alzheimer’s disease (AD), are challenging diseases for those affected with no cure and limited treatment options. Functional, human derived brain tissues that represent the diverse genetic background and cellular subtypes contributing to sporadic AD (sAD) are limited. Human stem cell derived brain organoids recapitulate some features of human brain cytoarchitecture and AD-like pathology, providing a tool for illuminating the relationship between AD pathology and neural cell dysregulation leading to cognitive decline. In this review, we explore current strategies for implementing brain organoids in the study of AD as well as the challenges associated with investigating age-related brain diseases using organoid models.