Chagas Disease: A Silent Threat for Dogs and Humans.

Chagas disease (CD) is a vector-borne Neglected Zoonotic Disease (NZD) caused by a flagellate protozoan, Trypanosoma cruzi, that affects various mammalian species across America, including humans and domestic animals. However, due to an increase in population movements and new routes of transmission, T. cruzi infection is presently considered a worldwide health concern, no longer restricted to endemic countries. Dogs play a major role in the domestic cycle by acting very efficiently as reservoirs and allowing the perpetuation of parasite transmission in endemic areas. Despite the significant progress made in recent years, still there is no vaccine against human and animal disease, there are few drugs available for the treatment of human CD, and there is no standard protocol for the treatment of canine CD. In this review, we highlight human and canine Chagas Disease in its different dimensions and interconnections. Dogs, which are considered to be the most important peridomestic reservoir and sentinel for the transmission of T. cruzi infection in a community, develop CD that is clinically similar to human CD. Therefore, an integrative approach, based on the One Health concept, bringing together the advances in genomics, immunology, and epidemiology can lead to the effective development of vaccines, new treatments, and innovative control strategies to tackle CD.

Exploiting Leishmania-Primed Dendritic Cells as Potential Immunomodulators of Canine Immune Response.

Dendritic cells (DCs) capture pathogens and process antigens, playing a crucial role in activating naïve T cells, bridging the gap between innate and acquired immunity. However, little is known about DC activation when facing Leishmania parasites. Thus, this study investigates in vitro activity of canine peripheral blood-derived DCs (moDCs) exposed to L. infantum and L. amazonensis parasites and their extracellular vesicles (EVs). L. infantum increased toll-like receptor 4 gene expression in synergy with nuclear factor κB activation and the generation of pro-inflammatory cytokines. This parasite also induced the expression of class II molecules of major histocompatibility complex (MHC) and upregulated co-stimulatory molecule CD86, which, together with the release of chemokine CXCL16, can attract and help in T lymphocyte activation. In contrast, L. amazonensis induced moDCs to generate a mix of pro- and anti-inflammatory cytokines, indicating that this parasite can establish a different immune relationship with DCs. EVs promoted moDCs to express class I MHC associated with the upregulation of co-stimulatory molecules and the release of CXCL16, suggesting that EVs can modulate moDCs to attract cytotoxic CD8+ T cells. Thus, these parasites and their EVs can shape DC activation. A detailed understanding of DC activation may open new avenues for the development of advanced leishmaniasis control strategies.

Hyaluronic acid-based gels for oral application: Comparison of in vitro effects on gingival cells and bacteria.

Purpose: This study aimed to evaluate the cytotoxic effects of different topical hyaluronic acid-based gels on human gingival fibroblasts and oral bacteria. Methods: Four different hyaluronate gels - Bexident® Aftas (BA), GUM® AftaClear (AfC), Gengigel®(G), Aloclair® Plus (AlC) and a chlorhexidine gel - Bexident®Gums(BG) were selected. Human gingival fibroblasts (HGF) were seeded in 48-well plates with different gel/culture medium concentrations (v/v%) and cell viability was evaluated at 1 and 3 days of culture. Cell morphology was assessed, and alterations graded according to ISO 10993-5:2009(E). Streptococcus oralis CECT 907T colony was, seed on 48-well plate or spread onto the blood agar plates and exposed to the different gel's concentration. The optical density (OD) was assessed, and the diameter of the inhibition zone was measured (mm). Results: BA and G elicited reduced HGF cytotoxicity, followed by AfC. AlC and BG were cytotoxic at concentrations up to 3% for all exposure times. PCM images of HGF showed moderate-to-severe alterations for AlC and BG and slight to mild changes, for BA, AfC and G. The highest antibacterial activity against S.oralis was observed on AlC and AfC, and no antibacterial activity was observed for BA and G. Inhibitory effect in sessile colonies was only observed in AlC and BG. Conclusions: AlC demonstrated superior antibacterial activities against S.oralis but a higher cytotoxic potential in HGF. BA and G presented the lowest cytotoxicity with little to no antibacterial effect. AfC demonstrated bacteriostatic effects and low cytotoxicity on HGF.

Cell Biological and Antibacterial Evaluation of a New Approach to Zirconia Implant Surfaces Modified with MTA.

Peri-implantitis continues to be one of the major reasons for implant failure. We propose a new approach to the incorporation of MTA into zirconia implant surfaces with Nd:YAG laser and investigate the biological and the microbiological responses of peri-implant cells. Discs of zirconia stabilized with yttria and titanium were produced according to the following four study groups: Nd:YAG laser-textured zirconia coated with MTA (Zr MTA), Nd:YAG laser-textured zirconia (Zr textured), polished zirconia discs, and polished titanium discs (Zr and Ti). Surface roughness was evaluated by contact profilometry. Human osteoblasts (hFOB), gingival fibroblasts (HGF hTERT) and S. oralis were cultured on discs. Cell adhesion and morphology, cell differentiation markers and bacterial growth were evaluated. Zr textured roughness was significantly higher than all other groups. SEM images reveal cellular adhesion at 1 day in all samples in both cell lines. Osteoblasts viability was lower in the Zr MTA group, unlike fibroblasts viability, which was shown to be higher in the Zr MTA group compared with the Zr textured group at 3 and 7 days. Osteocalcin and IL-8 secretion by osteoblasts were higher in Zr MTA. The Zr textured group showed higher IL-8 values released by fibroblasts. No differences in S. oralis CFUs were observed between groups. The present study suggests that zirconia implant surfaces coated with MTA induced fibroblast proliferation and osteoblast differentiation; however, they did not present antibacterial properties.

A metatranscriptomics strategy for efficient characterization of the microbiome in human tissues with low microbial biomass.

The high background of host RNA poses a major challenge to metatranscriptome analysis of human samples. Hence, metatranscriptomics has been mainly applied to microbe-rich samples, while its application in human tissues with low ratio of microbial to host cells has yet to be explored. Since there is no computational workflow specifically designed for the taxonomic and functional analysis of this type of samples, we propose an effective metatranscriptomics strategy to accurately characterize the microbiome in human tissues with a low ratio of microbial to host content. We experimentally generated synthetic samples with well-characterized bacterial and host cell compositions, and mimicking human samples with high and low microbial loads. These synthetic samples were used for optimizing and establishing the workflow in a controlled setting. Our results show that the integration of the taxonomic analysis of optimized Kraken 2/Bracken with the functional analysis of HUMAnN 3 in samples with low microbial content, enables the accurate identification of a large number of microbial species with a low false-positive rate, while improving the detection of microbial functions. The effectiveness of our metatranscriptomics workflow was demonstrated in synthetic samples, simulated datasets, and most importantly, human gastric tissue specimens, thus providing a proof of concept for its applicability on mucosal tissues of the gastrointestinal tract. The use of an accurate and reliable metatranscriptomics approach for human tissues with low microbial content will expand our understanding of the functional activity of the mucosal microbiome, uncovering critical interactions between the microbiome and the host in health and disease.

The Microbial Community Structure in the Rhizosphere of Theobroma cacao L. and Euterpe oleracea Mart. Is Influenced by Agriculture System in the Brazilian Amazon.

This study tested the hypothesis that cocoa monoculture (MS) and cocoa-açai agroforestry systems (AFS) may influence the microbial community structure and populations of plant growth-promoting bacteria (PGPR). Accordingly, the aim was to analyze the microbial community structure and PGPR populations in different agroecosystems in the Brazilian Amazon. To achieve this, the rhizosphere microbial community of cocoa and açai plants in both Amazonian seasons (dry and rainy) was analyzed using culture-dependent (PGPR screening) and -independent methods [PCR-DGGE based on rrs, alp, nifH gene, and intergenic region (ITS) of fungi]. Concerning PGPR screening, out of 48 isolated bacterial strains, 25% were capable of siderophore production, 29% of mineralized organic phosphate, 8% of inorganic phosphate solubilization, and 4% of indole acetic acid production. Moreover, 17% of isolates could inhibit the growth of various phytopathogenic fungi. Statistical analyses of DGGE fingerprints (p < 0.05) showed that bacterial and fungal community structures in the rhizosphere were influenced by the seasons, supporting the results of the physicochemical analysis of the environment. Furthermore, as hypothesized, microbial communities differed statistically when comparing the MS and AFS. These findings provide important insights into the influence of climate and cultivation systems on soil microbial communities to guide the development of sustainable agricultural practices.

A road map for the treatment of pediatric diffuse midline glioma.

Recent clinical trials for H3K27-altered diffuse midline gliomas (DMGs) have shown much promise. We present a consensus roadmap and identify three major barriers: (1) refinement of experimental models to include immune and brain-specific components; (2) collaboration among researchers, clinicians, and industry to integrate patient-derived data through sharing, transparency, and regulatory considerations; and (3) streamlining clinical efforts including biopsy, CNS-drug delivery, endpoint determination, and response monitoring. We highlight the importance of comprehensive collaboration to advance the understanding, diagnostics, and therapeutics for DMGs.

The Chromatin Remodeler CHD4 Sustains Ewing Sarcoma Cell Survival by Controlling Global Chromatin Architecture.

Ewing sarcoma is an aggressive cancer with a defective response to DNA damage leading to an enhanced sensitivity to genotoxic agents. Mechanistically, Ewing sarcoma is driven by the fusion transcription factor EWS-FLI1, which reprograms the tumor cell epigenome. The nucleosome remodeling and deacetylase (NuRD) complex is an important regulator of chromatin function, controlling both gene expression and DNA damage repair, and has been associated with EWS-FLI1 activity. Here, a NuRD-focused CRISPR/Cas9 inactivation screen identified the helicase CHD4 as essential for Ewing sarcoma cell proliferation. CHD4 silencing induced tumor cell death by apoptosis and abolished colony formation. Although CHD4 and NuRD colocalized with EWS-FLI1 at enhancers and super-enhancers, CHD4 promoted Ewing sarcoma cell survival not by modulating EWS-FLI1 activity and its oncogenic gene expression program but by regulating chromatin structure. CHD4 depletion led to a global increase in DNA accessibility and induction of spontaneous DNA damage, resulting in an increased susceptibility to DNA-damaging agents. CHD4 loss delayed tumor growth in vivo, increased overall survival, and combination with PARP inhibition by olaparib treatment further suppressed tumor growth. Collectively, these findings highlight the NuRD subunit CHD4 as a therapeutic target in Ewing sarcoma that can potentiate the antitumor activity of genotoxic agents. SIGNIFICANCE: CRISPR/Cas9 screening in Ewing sarcoma identifies a dependency on CHD4, which is crucial for the maintenance of chromatin architecture to suppress DNA damage and a promising therapeutic target for DNA damage repair-deficient malignancies.

Particle abrasion as a pre-bonding dentin surface treatment: A scoping review.

OBJECTIVE: This scoping review aims to assess the influence of air abrasion with aluminum oxide and bioactive glass on dentin bond strength. MATERIALS AND METHODS: An electronic search was conducted in three databases (PubMed, Cochrane Library, and Embase), on March 3rd, 2023, with previously identified MeSH Terms. A total of 1023 records were screened. Exclusion criteria include primary teeth, air abrasion of a substrate other than sound dentin, use of particles apart from aluminum oxide or bioactive glass, and studies in which bond strength was not assessed. RESULTS: Out of the 1023 records, title and abstract screening resulted in the exclusion of 895 and 67 studies, respectively, while full-text analysis excluded another 25 articles. In addition, 5 records were not included, as full texts could not be obtained after requesting the authors. Two cross-references were added. Thus, 33 studies were included in this review. It is important to emphasize the absence of standardization of air abrasion parameters. According to 63.6% of the studies, air abrasion does not influence dentin bond strength. Moreover, 30.3% suggest improving bonding performance, and 6.1% advocate a decrease. CONCLUSIONS: Air abrasion with aluminum oxide does not enhance or impair dentin bond strength. The available data on bioactive glass are limited, which hinders conclusive insights. CLINICAL SIGNIFICANCE: Dentin air abrasion is a widely applied technique nowadays, with numerous clinical applications. Despite the widespread adoption of this procedure, its potential impact on bonding performance requires a thorough analysis of the existing literature.

Targeting ß Cells with Cathelicidin Nanomedicines Improves Insulin Function and Pancreas Regeneration in Type 1 Diabetic Rats.

Type 1 diabetes (T1D) is an incurable condition with an increasing incidence worldwide, in which the hallmark is the autoimmune destruction of pancreatic insulin-producing ß cells. Cathelicidin-based peptides have been shown to improve ß cell function and neogenesis and may thus be relevant while developing T1D therapeutics. In this work, a cathelicidin-derived peptide, LLKKK18, was loaded in poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs), surface-functionalized with exenatide toward a GLP-1 receptor, aiming the ß cell-targeted delivery of the peptide. The NPs present a mean size of around 100 nm and showed long-term stability, narrow size distribution, and negative ζ-potential (-10 mV). The LLKKK18 association efficiency and loading were 62 and 2.9%, respectively, presenting slow and sustained in vitro release under simulated physiologic fluids. Glucose-stimulated insulin release in the INS-1E cell line was observed in the presence of the peptide. In addition, NPs showed a strong association with ß cells from isolated rat islets. After administration to diabetic rats, NPs induced a significant reduction of the hyperglycemic state, an improvement in the pancreatic insulin content, and glucose tolerance. Also remarkable, a considerable increase in the ß cell mass in the pancreas was observed. Overall, this novel and versatile nanomedicine showed glucoregulatory ability and can pave the way for the development of a new generation of therapeutic approaches for T1D treatment.

Permission to pass: on the role of p53 as a gatekeeper for aneuploidy.

Aneuploidy-the karyotype state in which the number of chromosomes deviates from a multiple of the haploid chromosome set-is common in cancer, where it is thought to facilitate tumor initiation and progression. However, it is poorly tolerated in healthy cells: during development and tissue homeostasis, aneuploid cells are efficiently cleared from the population. It is still largely unknown how cancer cells become, and adapt to being, aneuploid. P53, the gatekeeper of the genome, has been proposed to guard against aneuploidy. Aneuploidy in cancer genomes strongly correlates with mutations in TP53, and p53 is thought to prevent the propagation of aneuploid cells. Whether p53 also participates in preventing the mistakes in cell division that lead to aneuploidy is still under debate. In this review, we summarize the current understanding of the role of p53 in protecting cells from aneuploidy, and we explore the consequences of functional p53 loss for the propagation of aneuploidy in cancer.

Spanish cohort of VEXAS syndrome: clinical manifestations, outcome of treatments and novel evidences about UBA1 mosaicism.

BACKGROUND: The vacuoles, E1-enzyme, X linked, autoinflammatory and somatic (VEXAS) syndrome is an adult-onset autoinflammatory disease (AID) due to postzygotic UBA1 variants. OBJECTIVES: To investigate the presence of VEXAS syndrome among patients with adult-onset undiagnosed AID. Additional studies evaluated the mosaicism distribution and the circulating cytokines. METHODS: Gene analyses were performed by both Sanger and amplicon-based deep sequencing. Patients' data were collected from their medical charts. Cytokines were quantified by Luminex. RESULTS: Genetic analyses of enrolled patients (n=42) identified 30 patients carrying UBA1 pathogenic variants, with frequencies compatible for postzygotic variants. All patients were male individuals who presented with a late-onset disease (mean 67.5 years; median 67.0 years) characterised by cutaneous lesions (90%), fever (66.7%), pulmonary manifestations (66.7%) and arthritis (53.3%). Macrocytic anaemia and increased erythrocyte sedimentation rate and ferritin were the most relevant analytical abnormalities. Glucocorticoids ameliorated the inflammatory manifestations, but most patients became glucocorticoid-dependent. Positive responses were obtained when targeting the haematopoietic component of the disease with either decitabine or allogeneic haematopoietic stem cell transplantation. Additional analyses detected the UBA1 variants in both haematopoietic and non-haematopoietic tissues. Finally, analysis of circulating cytokines did not identify inflammatory mediators of the disease. CONCLUSION: Thirty patients with adult-onset AID were definitively diagnosed with VEXAS syndrome through genetic analyses. Despite minor interindividual differences, their main characteristics were in concordance with previous reports. We detected for the first time the UBA1 mosaicism in non-haematopoietic tissue, which questions the previous concept of myeloid-restricted mosaicism and may have conceptual consequences for the disease mechanisms.

A Case of Febrile Polyphagia With an Underlying Paraneoplastic Limbic Encephalitis.

A 57-year-old female with a history of malignant mixed Müllerian tumors of the uterus and ovaries developed a fever of unknown origin and neuropsychiatric symptoms. Her EEG showed slow activity in the left temporal region, and brain magnetic resonance imaging revealed limbic encephalitis, leading to the diagnosis of classic paraneoplastic limbic encephalitis (PLE). During our investigation into the underlying cause of the patient's condition, we conducted a PET-CT scan, which revealed the presence of several hypermetabolic lymph nodes. One of these lymph nodes underwent a biopsy, and the results confirmed the presence of metastatic cells, indicating the likelihood of carcinoma, most probably adenocarcinoma of the gynecological tract. PLE should be considered as one of the differential diagnoses in patients with a history of cancer and acute-to-subacute neuronal and psychiatric dysfunction.

Effects of endodontic irrigation solutions on structural, chemical, and mechanical properties of coronal dentin: A scoping review.

OBJECTIVE: This review aims to assess structural, chemical, and mechanical properties of coronal dentin after endodontic irrigation. MATERIALS AND METHODS: Reporting followed the PRISMA extension for scoping reviews. An electronic search was carried out in PubMed, Embase, and Cochrane Library. Records filtered by language and published up to November 4, 2022 were independently screened by two researchers. Studies evaluating structural, chemical, or mechanical properties of human permanent coronal dentin after irrigation within the scope of nonsurgical root canal treatment were included. Data were extracted regarding study type, sample description and size, experimental groups, outcome, evaluation method, and main findings. RESULTS: From the initial 1916 studies, and by adding 2 cross-references, 11 in vitro studies were included. Seven studies provide ultrastructural and/or chemical characterization, and six assessed microhardness and/or flexural strength. One percent to 8% sodium hypochlorite (NaOCl) and 1%-17% ethylenediaminetetraacetic acid (EDTA) were the most commonly tested solutions, with contact times of 2-240 min (NaOCl) and 1-1440 min (EDTA) being evaluated. CONCLUSIONS: Overall, the literature is consensual regarding the inevitable impact of NaOCl and chelating agents on coronal dentin, with both deproteinizing and decalcifying effects being concentration- and time-dependent. The alteration of mechanical parameters further confirmed the surface and subsurface ultrastructural and chemical changes. CLINICAL SIGNIFICANCE: Endodontic treatment success highly depends on restorative sealing. Understanding the result of exposing coronal dentin, the main substrate for bonding, to irrigants' action is crucial. The deproteinizing and decalcifying effects of NaOCl and chelating agents are both concentration- and time-dependent, causing surface and subsurface ultrastructural, chemical, and mechanical alterations.

Effectiveness of changing the color of darker teeth is potentiated by association with violet LED light.

BACKGROUND: The effectiveness of in-office bleaching protocols performed with violet LED light either combined with a bleaching agent containing 37% carbamide peroxide, or not, was determined by comparing teeth with different degrees of darkening. METHODOLOGY: Eighty bovine incisors were separated into groups of "light" teeth (luminosity greater than or equal to B3) and "dark" teeth (less than or equal to A3.5) to receive the protocols: HP - 35% hydrogen peroxide (Whiteness HP), CP - 37% carbamide peroxide (Whiteness SuperEndo), LED - violet LED light (Bright Max Whitening), CPLED - CP associated with the LED. For color analysis the CIEL*a*b* e WID, ΔEab, ΔE00 e ΔWID parameters were used. Data were analyzed using Mann-Whitney, Kruskal-Wallis, Dunn, Friedman or Nemenyi tests (α = 5%). RESULTS: HP and CP resulted in similar color change values (ΔEab, ΔE00 e ΔWID) for light and dark teeth (p > 0.05). Dark teeth showed better bleaching effectiveness (ΔEab, ΔE00 e ΔWID) than light teeth when CPLED was used (p < 0.05). LED showed color change that were below the limits of acceptability and perceptibility for ΔWID. CONCLUSION: light teeth are effectively bleached with the use of HP or CP, whereas dark teeth respond better to treatment with the CPLED protocol. Violet LED used alone did not show a satisfactory result.

Bio-Piezoelectric Ceramic Composites for Electroactive Implants-Biological Performance.

Barium titanate (BaTiO3) piezoelectric ceramic may be a potential alternative for promoting osseointegration due to its piezoelectric properties similar to bone electric potentials generated in loading function. In this sense, the aim of this in vitro study was to evaluate the cellular response of human osteoblasts and gingival fibroblasts as well as the impact on S. oralis when in contact with BaTiO3 functionalized zirconia implant surfaces with piezoelectric properties. Zirconia discs with BaTiO3 were produced and contact poling (piezo activation) was performed. Osteoblasts (hFOB 1.19), fibroblasts (HGF hTERT) and S. oralis were culture on discs. Cell viability and morphology, cell differentiation markers, bacterial adhesion and growth were evaluated. The present study suggests that zirconia composite surfaces with the addition of piezoelectric BaTiO3 are not cytotoxic to peri-implant cells. Also, they seem to promote a faster initial osteoblast differentiation. Moreover, these surfaces may inhibit the growth of S. oralis by acting as a bacteriostatic agent over time. Although the piezoelectric properties do not affect the cellular inflammatory profile, they appear to enable the initial adhesion of bacteria, however this is not significant over the entire testing period. Furthermore, the addition of non-poled BaTiO3 to zirconia may have a potential reduction effect on IL-6 mediated-inflammatory activity in fibroblasts.

A Challenging Case of Wilson's Disease.

Wilson's disease (WD) is an inherited disorder characterized by the accumulation of copper in various organs, particularly the liver, central nervous system, and cornea. The clinical presentation of WD can vary widely. Diagnosis requires a combination of clinical and biochemical findings. We present a case of a 20-year-old woman who presented to the Emergency Room with progressive motor decline. She exhibited characteristic neurological symptoms and signs, such as hypomimia, bradyphrenia, bradykinesia, dysarthria, sialorrhea, upper limb dystonia, and wing-beating tremor. Ophthalmological examination revealed corneal deposits known as Kayser-Fleischer rings. Laboratory investigations demonstrated low levels of ceruloplasmin and elevated serum copper. Brain MRI showed typical signs of copper deposition in the basal ganglia. The Leipzig criteria were used to confirm the diagnosis. Treatment with penicillamine and zinc acetate resulted in symptom improvement. This case highlights the diverse presentation of WD and the importance of early diagnosis and prompt treatment initiation.

Adenovirus Infection in a Kidney-Pancreatic Transplant Recipient: Case Report.

Adenovirus infection in transplant recipients may present from asymptomatic viremia to multisystemic involvement. Most frequently, it occurs in the first year after a kidney transplant, and it is secondary to the reactivation of latent disease. However, primary infection may occur, and disseminated disease is more common when related to primary infection. Kidney involvement may be confirmed by biopsy, although diagnosis may be presumptive. Reduction of immunosuppression and supportive care are important components of therapy. CASE DESCRIPTION: A 41-year-old female renal-pancreatic recipient 12 years before with chronic renal graft dysfunction and a functional pancreatic graft had a history of cytomegalovirus and polyoma virus infection 2 years after transplantation. She was taking tacrolimus, mycophenolate mofetil, and prednisolone. The patient was admitted after persistent uncharacteristic diarrhea 3 weeks before hospitalization without any relevant epidemiologic context. She was dehydrated, and the lab results showed worsened kidney function and leucocytosis. The viral culture revealed adenovirus. Vigorous hydration was implemented, and the mycophenolate mofetil dose was reduced. The patient was discharged, and renal function returned to previous values. DISCUSSION AND CONCLUSION: Adenovirus infection has a wide clinical presentation, and multisystemic involvement may occur in transplant recipients. Supportive care is paramount. The clinical features and viral culture confirm the diagnosis, although tissue samples and quantitative polymerase chain reaction may be required in more severe cases.