Rotenone inhibits embryonic chick myogenesis in a ROS-dependent mechanism.

Skeletal muscle function is highly dependent on the energy supply provided by mitochondria. Besides ATP production, mitochondria have several other roles, such as calcium storage, heat production, cell death signaling, autophagy regulation and redox state modulation. Mitochondrial function is crucial for skeletal muscle fiber formation. Disorders that affect mitochondria have a major impact in muscle development and function. Here we studied the role of mitochondria during chick skeletal myogenesis. We analyzed the intracellular distribution of mitochondria in myoblasts, fibroblasts and myotubes using Mitotracker labeling. Mitochondrial respiration was investigated in chick muscle cells. Our results show that (i) myoblasts and myotubes have more mitochondria than muscle fibroblasts; (ii) mitochondria are organized in long lines within the whole cytoplasm and around the nuclei of myotubes, while in myoblasts they are dispersed in the cytoplasm; (iii) the area of mitochondria in myotubes increases during myogenesis, while in myoblasts and fibroblasts there is a slight decrease; (iv) mitochondrial length increases in the three cell types (myoblasts, fibroblasts and myotubes) during myogenesis; (v) the distance of mitochondria to the nucleus increases in myoblasts and myotubes during myogenesis; (vi) Rotenone inhibits muscle fiber formation, while FCCP increases the size of myotubes; (vii) N-acetyl cysteine (NAC), an inhibitor of ROS formation, rescues the effects of Rotenone on muscle fiber size; and (viii) Rotenone induces the production of ROS in chick myogenic cells. The collection of our results suggests a role of ROS signaling in mitochondrial function during chick myogenesis.

A new hydroxyapatite-alginate-gelatin biocomposite favor bone regeneration in a critical-sized calvarial defect model.

This study aimed to evaluate the osteogenic potential of hydroxyapatite (HA), Alginate (Alg), and Gelatine (Gel) composite in a critical-size defect model in rats. Twenty-four male rats were divided into three groups: a negative control with no treatment (Control group), a positive control treated with deproteinized bovine bone mineral (DBBM group), and the experimental group treated with the new HA-Alg-Gel composite (HA-Alg-Gel group). A critical size defect (8.5mm) was made in the rat's calvaria, and the bone formation was evaluated by in vivo microcomputed tomography analysis (µCT) after 1, 15, 45, and 90 days. After 90 days, the animals were euthanized and histological and histomorphometric analyses were performed. A higher proportion of mineralized tissue/biomaterial was observed in the DBBM group when compared to the HA-Alg-Gel and Control groups in the µCT analysis during all analysis periods. However, no differences were observed in the mineralized tissue/biomaterial proportion observed on day 1 (immediate postoperative) in comparison to later periods of analysis in all groups. In the histomorphometric analysis, the HA-Alg-Gel and Control groups showed higher bone formation than the DBBM group. Moreover, in histological analysis, five samples of the HA-Alg-Gal group exhibited formed bone spicules adjacent to the graft granules against only two of eight samples in the DBBM group. Both graft materials ensured the maintenance of defect bone thickness, while a tissue thickness reduction was observed in the control group. In conclusion, this study demonstrated the osteoconductive potential of HA-Alg-Gel bone graft by supporting new bone formation around its particles.

Ancestral aneuploidy and stable chromosomal duplication resulting in differential genome structure and gene expression control in trypanosomatid parasites.

Aneuploidy is widely observed in both unicellular and multicellular eukaryotes, usually associated with adaptation to stress conditions. Chromosomal duplication stability is a tradeoff between the fitness cost of having unbalanced gene copies and the potential fitness gained from increased dosage of specific advantageous genes. Trypanosomatids, a family of protozoans that include species that cause neglected tropical diseases, are a relevant group to study aneuploidies. Their life cycle has several stressors that could select for different patterns of chromosomal duplications and/or losses, and their nearly universal use of polycistronic transcription increases their reliance on gene expansion/contraction, as well as post-transcriptional control as mechanisms for gene expression regulation. By evaluating the data from 866 isolates covering seven trypanosomatid genera, we have revealed that aneuploidy tolerance is an ancestral characteristic of trypanosomatids but has a reduced occurrence in a specific monophyletic clade that has undergone large genomic reorganization and chromosomal fusions. We have also identified an ancient chromosomal duplication that was maintained across these parasite's speciation, named collectively as the trypanosomatid ancestral supernumerary chromosome (TASC). TASC has most genes in the same coding strand, is expressed as a disomic chromosome (even having four copies), and has increased potential for functional variation, but it purges highly deleterious mutations more efficiently than other chromosomes. The evidence of stringent control over gene expression in this chromosome suggests that these parasites have adapted to mitigate the fitness cost associated with this ancient chromosomal duplication.

Ozone therapy improves early stages of osseointegration in ovariectomized rats.

OBJECTIVE: the aim of this study was to analyze the influence of ozone therapy (OZN) on peri-implant bone repair in critical bones by installing osseointegrated implants in the tibia of ovariectomized rats. METHODOLOGY: ovariectomy was performed on 30 Wistar rats, aged six months (Rattus novergicus), and, after 90 days, osseointegrated implants were installed in each tibial metaphysis. The study groups were divided into the animals that received intraperitoneal ozone at a concentration of 700 mcg/kg - OZ Group (n=15) - and a control group that received an intraperitoneal saline solution and, for this reason, was named the SAL group (n=15). The applications for both groups occurred during the immediate post-operative period on the 2nd, 4th, 6th, 8th, 10th, and 12th day post-surgery. At various stages (14, 42, and 60 days), the animals were euthanized, and tests were performed on their tibiae. These tests include histomorphometric and immunohistochemical analyses, computerized microtomography, sampling in light-cured resin for calcified sections, and confocal microscopy. The obtained data were then analyzed using One-way ANOVA and the Shapiro-Wilk, Kruskal-Wallis, and student t-tests (P<0.05). RESULTS: our findings indicate that the OZ group (3.26±0.20 mm) showed better cellular organization and bone neoformation at 14 days (SAL group, 0.90±1.42 mm) (P=0.001). Immunohistochemistry revealed that osteocalcin labeling was moderate in the OZ group and mild in the SAL group at 14 and 42 days post-surgery. The data from the analysis of calcified tissues (microtomography, histometric, and bone dynamism analysis) at 60 days showed no statistically significant differences between the groups (P=0.32). CONCLUSION: it was concluded that ozone therapy anticipated the initial phases of the peri-implant bone repair process.

BioDeepfuse: a hybrid deep learning approach with integrated feature extraction techniques for enhanced non-coding RNA classification.

The accurate classification of non-coding RNA (ncRNA) sequences is pivotal for advanced non-coding genome annotation and analysis, a fundamental aspect of genomics that facilitates understanding of ncRNA functions and regulatory mechanisms in various biological processes. While traditional machine learning approaches have been employed for distinguishing ncRNA, these often necessitate extensive feature engineering. Recently, deep learning algorithms have provided advancements in ncRNA classification. This study presents BioDeepFuse, a hybrid deep learning framework integrating convolutional neural networks (CNN) or bidirectional long short-term memory (BiLSTM) networks with handcrafted features for enhanced accuracy. This framework employs a combination of k-mer one-hot, k-mer dictionary, and feature extraction techniques for input representation. Extracted features, when embedded into the deep network, enable optimal utilization of spatial and sequential nuances of ncRNA sequences. Using benchmark datasets and real-world RNA samples from bacterial organisms, we evaluated the performance of BioDeepFuse. Results exhibited high accuracy in ncRNA classification, underscoring the robustness of our tool in addressing complex ncRNA sequence data challenges. The effective melding of CNN or BiLSTM with external features heralds promising directions for future research, particularly in refining ncRNA classifiers and deepening insights into ncRNAs in cellular processes and disease manifestations. In addition to its original application in the context of bacterial organisms, the methodologies and techniques integrated into our framework can potentially render BioDeepFuse effective in various and broader domains.

Antimanic-like effect of dipyridamole in the methylphenidate-induced hyperlocomotion.

BACKGROUND: Adenosinergic system has been implicated in the pathophysiology of bipolar disorder and drugs that affect adenosine neurotransmission have shown some efficacy as add-on therapy in manic patients. OBJECTIVE: Thus, the aim of the present study was to screen adenosinergic drugs for antimanic-like effect in methylphenidate (MPH)-induced hyperlocomotion in mice. METHODS: Male and female Swiss mice received a single allopurinol (50 and 200 mg/kg, ip), dipyridamole (20 mg/kg, ip), or inosine (50 mg/kg, ip) administration before an acute MPH challenge (5 mg/kg, sc). In experiments with repeated treatment, male mice received a daily administration of allopurinol (25 and 50 mg/kg, ip), dipyridamole (20 mg/kg, ip), or inosine (50 mg/kg, ip) for 14 days. Finally, pretreatment with aminophylline (2 mg/kg, sc), an unspecific adenosine receptor antagonist, was used to evaluate a putative adenosinergic mediation. Locomotor activity was measured in the automated activity chamber for 20 min. RESULTS: Acute and repeated dipyridamole reduced the increase in locomotor activity induced by MPH, while allopurinol and inosine had no effect. Aminophylline blocked the effect of dipyridamole in MPH-induced hyperlocomotion. CONCLUSION: The present results suggest that dipyridamole may have an antimanic-like effect through adenosine receptors and reinforce the proposal that the adenosine system may be an interesting target for new antimanic drugs.

Enhancing Weaned Piglet Health and Performance: The Role of Autolyzed Yeast (Saccharomyces cerevisiae) and ß-Glucans as a Blood Plasma Alternative in Diets.

The objective of this study was to evaluate the inclusion of the autolyzed yeast (AY) Saccharomyces cerevisiae with or without an immunomodulator (1,3/1,6 ß-glucans) as a total/partial substitute for blood plasma (BP) in the diet of post-weaning piglets; zootechnical performance, intestinal health and microbiota, immune responses and energy metabolism were assessed. A total of 240 castrated male and female piglets, with a mean age of 22 days and mean initial weight of 5.24 ± 0.82 kg, were randomly divided into blocks of four treatments with 12 replicates. The dietary inclusions were blood plasma (BP), autolyzed yeast (AY), autolyzed yeast + immunomodulator (AYI) and 50% BP and 50% AY (BPAY). In pre-initial phase II (29-35 days), piglets fed AY showed better feed conversion (FCR = 1.358) than the piglets in the BP (1.484), AYI (1.379) and BPAY (1.442) groups, i.e., 8.49% (0.126), 1.52% (0.021) and 4.50% (0.084), respectively (p = 0.0293). In the total period (21-42 days), better FCR was observed in the AYI (1.458) group, i.e., 4.64% (0.071), 1.15% (0.017) and 4.58% (0.070), than in the BP (1.529), AY (1.475) and BPAY (1.528) groups, respectively (p = 0.0150). In piglets fed AY (n = 3) and BPAY (n = 2), there was a reduction in the number of medications, i.e., 82.35% (-14n) and 88.23% (-15n), respectively (p = 0.0001), compared with that in the BP group (n = 17). In the AY group (73.83 mg/dL), AYI group (69.92 mg/dL), and BPAY group (69.58 mg/dL), piglets exhibited increases in triglyceride levels of 79.32%, 69.83%, and 69.00%, respectively, in comparison to those in the BP group, which had triglyceride levels of 41.17 mg/dL (p = 0.0400). The beta-hydroxybutyrate concentration in the AY group (79.96 ng/µL) was lower by 31.95%, 22.64%, and 5.89% compared to the BP group (117.50 ng/µL), AYI group (103.36 ng/µL), and BPAY group (84.67 ng/µL), respectively (p = 0.0072). In the AYI group, there was modulation of the microbiota, with an increase in the relative abundance of bacteria of the genera Lactobacillus, Collinsella and Bulleidia. AY, associated or not associated with an immunomodulator, is a potential substitute for BP in diets for piglets in the nursery phase, with positive effects on immune, metabolic, and intestinal microbial performance.

Nanoparticle-based DNA vaccine protects against SARS-CoV-2 variants in female preclinical models.

A safe and effective vaccine with long-term protection against SARS-CoV-2 variants of concern (VOCs) is a global health priority. Here, we develop lipid nanoparticles (LNPs) to provide safe and effective delivery of plasmid DNA (pDNA) and show protection against VOCs in female small animal models. Using a library of LNPs encapsulating unique barcoded DNA (b-DNA), we screen for b-DNA delivery after intramuscular administration. The top-performing LNPs are further tested for their capacity of pDNA uptake in antigen-presenting cells in vitro. The lead LNP is used to encapsulate pDNA encoding the HexaPro version of SARS-CoV-2 spike (LNP-HPS) and immunogenicity and protection is tested in vivo. LNP-HPS elicit a robust protective effect against SARS-CoV-2 Gamma (P.1), correlating with reduced lethality, decreased viral load in the lungs and reduced lung damage. LNP-HPS induce potent humoral and T cell responses against P.1, and generate high levels of neutralizing antibodies against P.1 and Omicron (B.1.1.529). Our findings indicate that the protective efficacy and immunogenicity elicited by LNP-HPS are comparable to those achieved by the approved COVID-19 vaccine from Biontech/Pfizer in animal models. Together, these findings suggest that LNP-HPS hold great promise as a vaccine candidate against VOCs.

Conhecimento dos profissionais intra-hospitalares acerca do suporte básico de vida em uma parada cardiorrespiratória / In-hospital professionals' knowledge about basic life support in cardiac arrest / Conocimiento de los profesionales intrahospitalarios sobre el soporte vital básico en la parada cardíaca

Objetivo: avaliar o efeito da intervenção educativa no conhecimento da equipe de enfermagem sobre o suporte básico de vida para o atendimento à parada cardiorrespiratória de adultos no ambiente intra-hospitalar. Método: estudo transversal com abordagem quantitativa, realizado com 25 profissionais de enfermagem em dois hospitais de região oeste de Santa Catarina - Brasil. Avaliou-se por meio da aplicação de um pré-teste, intervenção educativa e pós-teste. Resultados: houve aumento significativo no conhecimento dos profissionais. O hospital A obteve a média de acertos de 7,23 no pré-teste, elevando para 11,33 no pós-teste, com valor de p ≤ 0,0001. Já o hospital B pontuou 6,07 no pré-teste, progredindo para 11,15 no pós-teste, valor de p ≤ 0,0006. Conclusão: a intervenção realizada demonstrou ser uma estratégia eficaz, visto que os resultados pré-teste demostravam déficit significativo de conhecimento, e após a intervenção educativa, mostraram melhoria na maioria dos itens avaliados em relação ao atendimento específico.

Objective: to evaluate the effect of an educational intervention on the nursing team's knowledge about basic life support for adult cardiac arrest care in the in-hospital environment. Method: cross-sectional study with a quantitative approach, carried out with 25 nursing professionals in two hospitals in the western region of Santa Catarina - Brazil. A pre-test, educational intervention and post-test were applied. Results: there was a significant increase in the professionals' knowledge. Hospital A had a mean score of 7.23 in the pre-test, increasing to 11.33 in the post-test, with p-value ≤ 0.0001. Hospital B scored 6.07 in the pre-test, increasing to 11.15 in the post-test, p-value ≤ 0.0006. Conclusion: the intervention proved to be an effective strategy, since the pre-test results showed significant knowledge deficit, and after the educational intervention, showed improvement in most of the items evaluated in relation to specific care.

Objetivos:evaluar el efecto de una intervención educativa en el conocimiento del equipo de enfermería sobre el soporte vital básico para la atención del paro cardíaco del adulto en el ambiente intrahospitalario. Método: estudio transversal con abordaje cuantitativo, realizado con 25 profesionales de enfermería en dos hospitales de la región oeste de Santa Catarina - Brasil. Se aplicó un pre-test, una intervención educativa y un post-test. Resultados: hubo un aumento significativo de los conocimientos de los profesionales. El Hospital A obtuvo una puntuación media de 7,23 en el pre-test, aumentando a 11,33 en el post-test, con valor p ≤ 0,0001. El Hospital B obtuvo una puntuación de 6,07 en el pre-test, aumentando a 11,15 en el post-test, con valor p ≤ 0,0006. Conclusión: una intervención realizada demostró ser una estrategia eficaz, visto que os resultados previos demostraron un déficit significativo de conhecimento, y después de una intervención educativa, mostraron una mejoría na maioria dos itens avaliados em relação ao atendimento específico.

Ozone therapy improves early stages of osseointegration in ovariectomized rats

Abstract Objective the aim of this study was to analyze the influence of ozone therapy (OZN) on peri-implant bone repair in critical bones by installing osseointegrated implants in the tibia of ovariectomized rats. Methodology ovariectomy was performed on 30 Wistar rats, aged six months (Rattus novergicus), and, after 90 days, osseointegrated implants were installed in each tibial metaphysis. The study groups were divided into the animals that received intraperitoneal ozone at a concentration of 700 mcg/kg — OZ Group (n=15) — and a control group that received an intraperitoneal saline solution and, for this reason, was named the SAL group (n=15). The applications for both groups occurred during the immediate post-operative period on the 2nd, 4th, 6th, 8th, 10th, and 12th day post-surgery. At various stages (14, 42, and 60 days), the animals were euthanized, and tests were performed on their tibiae. These tests include histomorphometric and immunohistochemical analyses, computerized microtomography, sampling in light-cured resin for calcified sections, and confocal microscopy. The obtained data were then analyzed using One-way ANOVA and the Shapiro-Wilk, Kruskal-Wallis, and student t-tests (P<0.05). Results our findings indicate that the OZ group (3.26±0.20 mm) showed better cellular organization and bone neoformation at 14 days (SAL group, 0.90±1.42 mm) (P=0.001). Immunohistochemistry revealed that osteocalcin labeling was moderate in the OZ group and mild in the SAL group at 14 and 42 days post-surgery. The data from the analysis of calcified tissues (microtomography, histometric, and bone dynamism analysis) at 60 days showed no statistically significant differences between the groups (P=0.32). Conclusion it was concluded that ozone therapy anticipated the initial phases of the peri-implant bone repair process.

Trypanosoma sp. infection in Boa constrictor snakes: morphological, hematological, clinical biochemistry, molecular, and phylogenetic characteristics.

There are few reports of Trypanosoma in snakes, as well as little information about its pathogenicity in these animals. Thus, the present study aimed to characterize Trypanosoma found in Boa constrictor snakes, to verify the influence of the parasitism on hematological and clinical biochemistry parameters, and to perform a phylogenetic study of the isolates. Blood samples from sixty-one boas were analyzed for the presence of trypanosomatids and by hematological and clinical biochemistry assays. The flagellates that were found in this analysis were used for cell culture, morphometry, and molecular analysis. Later, molecular typing phylogenetic studies were performed. Nine positive animals (14.75%) were identified by microscopy analysis. The hematological results showed that parasitized animals presented significantly lower levels of packed cell volume, hemoglobin, mean corpuscular volume, and mean corpuscular hemoglobin. In the leukogram, eosinophils and heterophils counts were higher in parasitized animals. Considering the molecular analyses, the isolates presented a higher identity of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the 18S small subunit ribosomal RNA (SSU rRNA) gene fragments with Trypanosoma serpentis. The phylogenetic tree, using the GAPDH, clustered all isolates with T. serpentis and Trypanosoma cascavelli. This is the first description of T. serpentis parasitizing boas and of the clinical changes caused by trypanosomatid infection in snakes.

Predictability of single versus double-barrel vascularized fibula flaps and dental implants in mandibular reconstructions: A systematic review and meta-analysis of prospective studies.

STATEMENT OF PROBLEM: Patients with vascularized bone flaps from the fibula have reduced bone height, in which case a higher prosthetic abutment is needed for their implant-supported prosthesis. Although the double-flap technique seems promising, systematic reviews and meta-analyses of prospective studies are lacking. PURPOSE: The purpose of this systematic review and meta-analysis was to evaluate the grafted areas of single barrel fibular flaps (SBFF) and double-barrel fibular flaps (DBFF) by considering failure rates, dental implant complications, and bone union at the osteotomy sites. MATERIAL AND METHODS: A systematic review and meta-analysis was performed in accordance with the preferred reporting items for systematic reviews and meta-analyses (PRISMA) statement, population, intervention, control, and outcomes (PICO) question, and the National Health and Medical Research Council scales. The event rate of complications and failures was calculated with a confidence interval (CI) of 95%. RESULTS: A total of 13 prospective studies with 441 participants and 330 graft sites were identified. A total of 235 participants had SBFF with 445 implants, and 95 had DBFF with 164 implants. The overall combined graft failure rates were 4.2% for SBFF and 3.2% for DBFF. The complication rate was 10% for SBFF and 1.9% for DBFF. Implant failure was at 4.7% in the SBFF group and 3.4% in the DBFF group. CONCLUSIONS: Complication rates and implant failures were similar for SBFF and DBFF. Therefore, for long-term oral rehabilitation, both SBFF and DBFF are suitable procedures for mandibular reconstruction.

Delivery of Plasmid DNA by Ionizable Lipid Nanoparticles to Induce CAR Expression in T Cells.

Introduction: Chimeric antigen receptor (CAR) cell therapy represents a hallmark in cancer immunotherapy, with significant clinical results in the treatment of hematological tumors. However, current approved methods to engineer T cells to express CAR use viral vectors, which are integrative and have been associated with severe adverse effects due to constitutive expression of CAR. In this context, non-viral vectors such as ionizable lipid nanoparticles (LNPs) arise as an alternative to engineer CAR T cells with transient expression of CAR. Methods: Here, we formulated a mini-library of LNPs to deliver pDNA to T cells by varying the molar ratios of excipient lipids in each formulation. LNPs were characterized and screened in vitro using a T cell line (Jurkat). The optimized formulation was used ex vivo to engineer T cells derived from human peripheral blood mononuclear cells (PBMCs) for the expression of an anti-CD19 CAR (CAR-CD19BBz). The effectiveness of these CAR T cells was assessed in vitro against Raji (CD19+) cells. Results: LNPs formulated with different molar ratios of excipient lipids efficiently delivered pDNA to Jurkat cells with low cytotoxicity compared to conventional transfection methods, such as electroporation and lipofectamine. We show that CAR-CD19BBz expression in T cells was transient after transfection with LNPs. Jurkat cells transfected with our top-performing LNPs underwent activation when exposed to CD19+ target cells. Using our top-performing LNP-9-CAR, we were able to engineer human primary T cells to express CAR-CD19BBz, which elicited significant specific killing of CD19+ target cells in vitro. Conclusion: Collectively, our results show that LNP-mediated delivery of pDNA is a suitable method to engineer human T cells to express CAR, which holds promise for improving the production methods and broader application of this therapy in the future.

The AnimalAssociatedMetagenomeDB reveals a bias towards livestock and developed countries and blind spots in functional-potential studies of animal-associated microbiomes.

BACKGROUND: Metagenomic data can shed light on animal-microbiome relationships and the functional potential of these communities. Over the past years, the generation of metagenomics data has increased exponentially, and so has the availability and reusability of data present in public repositories. However, identifying which datasets and associated metadata are available is not straightforward. We created the Animal-Associated Metagenome Metadata Database (AnimalAssociatedMetagenomeDB - AAMDB) to facilitate the identification and reuse of publicly available non-human, animal-associated metagenomic data, and metadata. Further, we used the AAMDB to (i) annotate common and scientific names of the species; (ii) determine the fraction of vertebrates and invertebrates; (iii) study their biogeography; and (iv) specify whether the animals were wild, pets, livestock or used for medical research. RESULTS: We manually selected metagenomes associated with non-human animals from SRA and MG-RAST.  Next, we standardized and curated 51 metadata attributes (e.g., host, compartment, geographic coordinates, and country). The AAMDB version 1.0 contains 10,885 metagenomes associated with 165 different species from 65 different countries. From the collected metagenomes, 51.1% were recovered from animals associated with medical research or grown for human consumption (i.e., mice, rats, cattle, pigs, and poultry). Further, we observed an over-representation of animals collected in temperate regions (89.2%) and a lower representation of samples from the polar zones, with only 11 samples in total. The most common genus among invertebrate animals was Trichocerca (rotifers). CONCLUSION: Our work may guide host species selection in novel animal-associated metagenome research, especially in biodiversity and conservation studies. The data available in our database will allow scientists to perform meta-analyses and test new hypotheses (e.g., host-specificity, strain heterogeneity, and biogeography of animal-associated metagenomes), leveraging existing data. The AAMDB WebApp is a user-friendly interface that is publicly available at https://webapp.ufz.de/aamdb/ .

The Interplay of Raloxifene and Sonochemical Bio-Oss in Early Maxillary Sinus Bone Regeneration: A Histological and Immunohistochemical Analysis in Rabbits.

The study aimed to assess the efficacy of using Raloxifene with ultrasonic processing to enhance Bio-Oss®, a bone graft substitute, for maxillary sinus bone height reconstruction. A total of 24 rabbit maxillary sinuses were distributed into three groups, each receiving different treatments: Bio-Oss® only, sonicated Bio-Oss, and sonicated Bio-Oss® with Raloxifene. Surgical procedures and subsequent histomorphometric and immunohistochemistry analyses were conducted to evaluate the bone formation, connective tissue, and remaining biomaterial, as well as the osteoblastic differentiation and maturation of collagen fibers. Results indicated that the sonicated Bio-Oss® and Bio-Oss® groups showed similar histological behavior and bone formation, but the Raloxifene group displayed inflammatory infiltrate, low bone formation, and disorganized connective tissue. The statistical analysis confirmed significant differences between the groups in terms of bone formation, connective tissue, and remaining biomaterial. In conclusion, the study found that while sonicated Bio-Oss® performed comparably to Bio-Oss® alone, the addition of Raloxifene led to an unexpected delay in bone repair. The findings stress the importance of histological evaluation for accurate bone repair assessment and the necessity for further investigation into the local application of Raloxifene. Future research may focus on optimizing bone substitutes with growth factors to improve bone repair.

Voltammetric Sensing of Nifedipine Using a Glassy Carbon Electrode Modified with Carbon Nanofibers and Gold Nanoparticles.

Nifedipine, a widely utilized medication, plays a crucial role in managing blood pressure in humans. Due to its global prevalence and extensive usage, close monitoring is necessary to address this widespread concern effectively. Therefore, the development of an electrochemical sensor based on a glassy carbon electrode modified with carbon nanofibers and gold nanoparticles in a Nafion® film was performed, resulting in an active electrode surface for oxidation of the nifedipine molecule. This was applied, together with a voltammetric methodology, for the analysis of nifedipine in biological and environmental samples, presenting a linear concentration range from 0.020 to 2.5 × 10-6 µmol L-1 with a limit of detection 2.8 nmol L-1. In addition, it presented a good recovery analysis in the complexity of the samples, a low deviation in the presence of interfering potentials, and good repeatability between measurements.

SOFT TISSUE SARCOMA - SANTA CASA DE SÃO PAULO EXPERIENCE FROM 2006 TO 2019.

Objective: To conduct an epidemiologic review, analyzing treatment, evolution, and survival of soft tissue sarcomas. Methods: Retrospective study based on medical records of patient with STS treated by the Orthopedic Oncology Group at the Santa Casa de São Paulo, from 2006 to 2019. Data from 121 patients were analyzed according to age, sex, histological type, tumor location, treatment, previous surgery in a non-specialized service, local recurrences, lung metastases, and survival analysis. Results: The most frequent location was the thigh. Patients who underwent surgery with a non-specialized group had higher rates of local recurrence and those with pulmonary metastasis had a lower survival rate. Conclusion: STS can occur at any age and the prevalence of the histological type depends on the patients' age group. Level of Evidence II, Prognostic Study.

Objetivo: Conduzir uma avaliação epidemiológica analisando tratamento, evolução e sobrevida dos sarcomas de partes moles (SPMs). Métodos: Estudo retrospectivo de prontuários de pacientes com SPM tratados pelo Grupo de Oncologia Ortopédica da Santa Casa de Misericórdia de São Paulo, no período de 2006 a 2019. Foram analisados os dados de 121 pacientes referentes a idade, sexo, tipo histológico, localização do tumor, tratamento, cirurgia prévia em serviço não especializado, presença de recidivas, metástases pulmonares e análise de sobrevida. Resultados: A localização mais frequente foi a coxa. Verificou-se que pacientes que realizaram cirurgia com grupo não especializado tiveram maiores índices de recidiva local, e aqueles com metástase pulmonar tiveram menor sobrevida. Conclusão: Os SPMs podem ocorrer em qualquer idade, e a prevalência do tipo histológico depende da faixa etária dos pacientes. Nível de Evidência II, Estudo Prognóstico.

Simultaneous Determination of Uric Acid and Caffeine by Flow Injection Using Multiple-Pulse Amperometry.

The present study reports the development and application of a flow injection analysis (FIA) system for the simultaneous determination of uric acid (UA) and caffeine (CAF) using cathodically pretreated boron-doped diamond electrode (CPT-BDD) and multiple-pulse amperometry (MPA). The electrochemical profiles of UA and CAF were analyzed via cyclic voltammetry in the potential range of 0.20-1.7 V using 0.10 mol L-1 H2SO4 solution as supporting electrolyte. Under optimized conditions, two oxidation peaks at potentials of 0.80 V (UA) and 1.4 V (CAF) were observed; the application of these potentials using multiple-pulse amperometry yielded concentration linear ranges of 5.0 × 10-8-2.2 × 10-5 mol L-1 (UA) and 5.0 × 10-8-1.9 × 10-5 mol L-1 (CAF) and limits of detection of 1.1 × 10-8 and 1.3 × 10-8 mol L-1 for UA and CAF, respectively. The proposed method exhibited good repeatability and stability, and no interference was detected in the electrochemical signals of UA and CAF in the presence of glucose, NaCl, KH2PO4, CaCl2, urea, Pb, Ni, and Cd. The application of the FIA-MPA method for the analysis of environmental samples resulted in recovery rates ranging between 98 and 104%. The results obtained showed that the BDD sensor exhibited a good analytical performance when applied for CAF and UA determination, especially when compared to other sensors reported in the literature.

Generation and Manipulation of Rat Intestinal Organoids.

When using organoids to assess physiology and cell fate decisions, it is important to use a model that closely recapitulates in vivo contexts. Accordingly, patient-derived organoids are used for disease modeling, drug discovery, and personalized treatment screening. Mouse intestinal organoids are commonly utilized to understand aspects of both intestinal function/physiology and stem cell dynamics/fate decisions. However, in many disease contexts, rats are often preferred over mice as a model due to their greater physiological similarity to humans in terms of disease pathophysiology. The rat model has been limited by a lack of genetic tools available in vivo, and rat intestinal organoids have proven fragile and difficult to culture long-term. Here, we build upon previously published protocols to robustly generate rat intestinal organoids from the duodenum and jejunum. We provide an overview of several downstream applications utilizing rat intestinal organoids, including functional swelling assays, whole mount staining, the generation of 2D enteroid monolayers, and lentiviral transduction. The rat organoid model provides a practical solution to the need of the field for an in vitro model which retains physiological relevance to humans, can be quickly genetically manipulated, and is easily obtained without the barriers involved in procuring human intestinal organoids.

Different culture media and purification methods unveil the core proteome of Propionibacterium freudenreichii-derived extracellular vesicles.

Bacterial extracellular vesicles (EVs) are natural lipidic nanoparticles implicated in intercellular communication. Although EV research focused mainly on pathogens, the interest in probiotic-derived EVs is now rising. One example is Propionibacterium freudenreichii, which produces EVs with anti-inflammatory effects on human epithelial cells. Our previous study with P. freudenreichii showed that EVs purified by size exclusion chromatography (SEC) displayed variations in protein content according to bacterial growth conditions. Considering these content variations, we hypothesized that a comparative proteomic analysis of EVs recovered in different conditions would elucidate whether a representative vesicular proteome existed, possibly providing a robust proteome dataset for further analysis. Therefore, P. freudenreichii was grown in two culture media, and EVs were purified by sucrose density gradient ultracentrifugation (UC). Microscopic and size characterization confirmed EV purification, while shotgun proteomics unveiled that they carried a diverse set of proteins. A comparative analysis of the protein content of UC- and SEC-derived EVs, isolated from cultures either in UF (cow milk ultrafiltrate medium) or YEL (laboratory yeast extract lactate medium), showed that EVs from all these conditions shared 308 proteins. This EV core proteome was notably enriched in proteins related to immunomodulation. Moreover, it showed distinctive features, including highly interacting proteins, compositional biases for some specific amino acids, and other biochemical parameters. Overall, this work broadens the toolset for the purification of P. freudenreichii-derived EVs, identifies a representative vesicular proteome, and enumerates conserved features in vesicular proteins. These results hold the potential for providing candidate biomarkers of purification quality, and insights into the mechanisms of EV biogenesis and cargo sorting.