New Surgical Model for Bone–Muscle Injury Reveals Age and Gender-Related Healing Patterns in the 5 Lipoxygenase (5LO) Knockout Mouse

Signaling lipid mediators released from 5 lipoxygenase (5LO) pathways influence both bone and muscle cells, interfering in their proliferation and differentiation capacities. A major limitation to studying inflammatory signaling pathways in bone and muscle healing is the inadequacy of available animal models. We developed a surgical injury model in the vastus lateralis (VL) muscle and femur in 129/SvEv littermates mice to study simultaneous musculoskeletal (MSK) healing in male and female, young (3 months) and aged (18 months) WT mice compared to mice lacking 5LO (5LOKO). MSK defects were surgically created using a 1-mm punch device in the VA muscle followed by a 0.5-mm round defect in the femur. After days 7 and 14 post-surgery, the specimens were removed for microtomography (microCT), histopathology, and immunohistochemistry analyses. In addition, non-injured control skeletal muscles along with femur and L5 vertebrae were analyzed. Bones were microCT phenotyped, revealing that aged female WT mice presented reduced BV/TV and trabecular parameters compared to aged males and aged female 5LOKO mice. Skeletal muscles underwent a customized targeted lipidomics investigation for profiling and quantification of lipid signaling mediators (LMs), evidencing age, and gender related-differences in aged female 5LOKO mice compared to matched WT. Histological analysis revealed a suitable bone-healing process with osteoid deposition at day 7 post-surgery, followed by woven bone at day 14 post-surgery, observed in all young mice. Aged WT females displayed increased inflammatory response at day 7 post-surgery, delayed bone matrix maturation, and increased TRAP immunolabeling at day 14 post-surgery compared to 5LOKO females. Skeletal muscles of aged animals showed higher levels of inflammation in comparison to young controls at day 14 post-surgery; however, inflammatory process was attenuated in aged 5LOKO mice compared to aged WT. In conclusion, this new model shows that MSK healing is influenced by age, gender, and the 5LO pathway, which might serve as a potential target to investigate therapeutic interventions and age-related MSK diseases. Our new model is suitable for bone–muscle crosstalk studies

Designing Logic Tensor Networks for Visual Sudoku puzzle classification

Given the increasing importance of the neurosymbolic (NeSy) approach in artificial intelligence, there is a growing interest in studying benchmarks specifically designed to emphasize the ability of AI systems to combine low-level representation learning with high-level symbolic reasoning. One such recent benchmark is Visual Sudoku Puzzle Classification, that combines visual perception with relational constraints. In this work, we investigate the application of Logic Tensork Networks (LTNs) to the Visual Sudoku Classification task and discuss various alternatives in terms of logical constraint formulation, integration with the perceptual module and training procedure

Importância da vacinação contra o papilomavírus humano em um assentamento rural em Terenos, Mato Grosso do Sul

OBJECTIVE: To understand health professionals’ perceptions about vaccination against human papillomavirus (HPV) in the Santa Mônica rural settlement in Terenos, Mato Grosso do Sul. METHODS: Quantitative and qualitative methodologies, consultations on vaccination cards, records of community health agents and the focus group technique were used. The main factors of hesitation and vaccine refusal were analyzed, as well as the health team’s strategies for the process of immunization against HPV, from June to August 2018. RESULTS: Of 121 children and adolescents, 81 (66.94%) received the complete vaccination schedule. Complete vaccination coverage for women was 73.17% (60/82) and for men, 53.8% (21/39). It was observed that, although strategies are adopted for vaccine promotion, such as mobile actions, the public is resistant due to superficial knowledge about the vaccine and its use in an early age group, showing itself to be susceptible to the negative influence of the media and to society’s taboos. In addition, difficulties regarding the use of the Unified Health System card and the shortage of professionals were also observed. CONCLUSION: The results explain the immunization coverage below the target and reinforce the need to strengthen the family health strategy, as well as the permanent and continuing education of professionals, in order to increase parental confidence and adherence to vaccination.OBJETIVO: Compreender as percepções dos profissionais de saúde acerca da vacinação contra o papilomavírus humano (HPV) no Complexo de assentamentos Santa Mônica, em Terenos, Mato Grosso do Sul. MÉTODOS: Foram utilizadas metodologias quanti-qualitativas, consultas em cartões vacinais, registros de agente comunitários de saúde e a técnica de grupo focal. Foram analisados os principais fatores de hesitação e recusa vacinal, bem como as estratégias da equipe de saúde para o processo de imunização contra o HPV, de junho a agosto de 2018. RESULTADOS: De 121 crianças e adolescentes, 81 (66,94%) receberam o esquema vacinal completo. A cobertura vacinal completa feminina foi de 73,17% (60/82) e a masculina de 53,8% (21/39). Observou-se que, embora sejam adotadas estratégias para a promoção da vacina, como ações volantes, o público encontra-se resistente devido ao conhecimento superficial sobre a vacina e sua utilização em faixa etária precoce, mostrando-se suscetível à influência negativa da mídia e aos tabus da sociedade. Além disso, dificuldades quanto ao uso do cartão do Sistema Único de Saúde e a escassez de profissionais também foram observadas. CONCLUSÃO: Os resultados justificam a cobertura vacinal abaixo da meta e reforçam a necessidade de fortalecimento da estratégia de saúde da família, bem como da educação permanente e continuada dos profissionais, a fim de aumentar a confiança dos pais e a adesão à vacinação

Compatibility of Azospirillum brasilense with Pesticides Used for Treatment of Maize Seeds

Seed treatment with chemical pesticides is commonly used as an initial plant protection procedure against pests and diseases. However, the use of such chemicals may impair the survival and performance of beneficial microorganisms introduced via inoculants, such as the plant growth-promoting bacterium Azospirillum brasilense. We assessed the compatibility between the most common pesticide used in Brazil for the treatment of maize seeds, composed of two fungicides, and one insecticide, with the commercial strains Ab-V5 and Ab-V6 of A. brasilense, and evaluated the impacts on initial plant development. The toxicity of the pesticide to A. brasilense was confirmed, with an increase in cell mortality after only 24 hours of exposure in vitro. Seed germination and seedling growth were not affected neither by the A. brasilense nor by the pesticide. However, under greenhouse conditions, the pesticide affected root volume and dry weight and root-hair incidence, but the toxicity was alleviated by the inoculation with A. brasilense for the root volume and root-hair incidence parameters. In maize seeds inoculated with A. brasilense, the pesticide negatively affected the number of branches, root-hair incidence, and root-hair length. Therefore, new inoculant formulations with cell protectors and the development of compatible pesticides should be searched to guarantee the benefits of inoculation with plant growth-promoting bacteria

PSMA-Specific CAR-Engineered T Cells Eradicate Disseminated Prostate Cancer in Preclinical Models

Immunology-based interventions have been proposed as a promising curative chance to effectively attack postoperative minimal residual disease and distant metastatic localizations of prostate tumors. We developed a chimeric antigen receptor (CAR) construct targeting the human prostate-specific membrane antigen (hPSMA), based on a novel and high affinity specific mAb. As a transfer method, we employed last-generation lentiviral vectors (LV) carrying a synthetic bidirectional promoter capable of robust and coordinated expression of the CAR molecule, and a bioluminescent reporter gene to allow the tracking of transgenic T cells after in vivo adoptive transfer. Overall, we demonstrated that CAR-expressing LV efficiently transduced short-term activated PBMC, which in turn were readily stimulated to produce cytokines and to exert a relevant cytotoxic activity by engagement with PSMA+ prostate tumor cells. Upon in vivo transfer in tumor-bearing mice, CAR-transduced T cells were capable to completely eradicate a disseminated neoplasia in the majority of treated animals, thus supporting the translation of such approach in the clinical setting

Virtual Drug Repositioning as a Tool to Identify Natural Small Molecules That Synergize with Lumacaftor in F508del-CFTR Binding and Rescuing

Cystic fibrosis is a hereditary disease mainly caused by the deletion of the Phe 508 (F508del) of the cystic fibrosis transmembrane conductance regulator (CFTR) protein that is thus withheld in the endoplasmic reticulum and rapidly degraded by the ubiquitin/proteasome system. Cystic fibrosis remains a potentially fatal disease, but it has become treatable as a chronic condition due to some CFTR-rescuing drugs that, when used in combination, increase in their therapeutic effect due to a synergic action. Also, dietary supplementation of natural compounds in combination with approved drugs could represent a promising strategy to further alleviate cystic fibrosis symptoms. On these bases, we screened by in silico drug repositioning 846 small synthetic or natural compounds from the AIFA database to evaluate their capacity to interact with the highly druggable lumacaftor binding site of F508del-CFTR. Among the identified hits, nicotinamide (NAM) was predicted to accommodate into the lumacaftor binding region of F508del-CFTR without competing against the drug but rather stabilizing its binding. The effective capacity of NAM to bind F508del-CFTR in a lumacaftor-uncompetitive manner was then validated experimentally by surface plasmon resonance analysis. Finally, the capacity of NAM to synergize with lumacaftor increasing its CFTR-rescuing activity was demonstrated in cell-based assays. This study suggests the possible identification of natural small molecules devoid of side effects and endowed with the capacity to synergize with drugs currently employed for the treatment of cystic fibrosis, which hopefully will increase the therapeutic efficacy with lower doses

A Mutant-p53/Smad Complex Opposes p63 to Empower TGFβ-Induced Metastasis

SummaryTGFβ ligands act as tumor suppressors in early stage tumors but are paradoxically diverted into potent prometastatic factors in advanced cancers. The molecular nature of this switch remains enigmatic. Here, we show that TGFβ-dependent cell migration, invasion and metastasis are empowered by mutant-p53 and opposed by p63. Mechanistically, TGFβ acts in concert with oncogenic Ras and mutant-p53 to induce the assembly of a mutant-p53/p63 protein complex in which Smads serve as essential platforms. Within this ternary complex, p63 functions are antagonized. Downstream of p63, we identified two candidate metastasis suppressor genes associated with metastasis risk in a large cohort of breast cancer patients. Thus, two common oncogenic lesions, mutant-p53 and Ras, selected in early neoplasms to promote growth and survival, also prefigure a cellular set-up with particular metastasis proclivity by TGFβ-dependent inhibition of p63 function