RESUMO
Fluazuron is a novel veterinary pour-on antitick formulation which can be applied simultaneously with bovine reproduction management strategies. Considering the economic importance of the livestock industry in many countries, it is important to know whether antiparasitics such as fluazuron may cause embryonic loss. The aim of this study was to evaluate the toxicological effect of fluazuron on bovine oocytes during in vitro maturation. The best fluazuron concentrations were determined in a preliminary experiment on Chinese hamster ovary (CHO)-K1 cells and further used to compare fluazuron toxicity in both study models. Results of the annexin V and alkaline single cell gel electrophoresis assays demonstrated that fluazuron caused cytotoxicity and genotoxicity in bovine cumulus cells at all the concentrations tested (50, 75 and 100 µg fluazuron/mL). The evaluation of cortical granules and mitochondria distribution showed that cytoplasmic maturation was not affected by fluazuron treatment. However, a decrease in metaphase II + polar body, degenerate oocytes as well as disorganized chromatin in polar body were observed at all concentrations tested. Whereas the fertilization process was not altered by 50 µg/mL fluazuron, the embryo development rate decreased significantly. No significant differences were observed in any of the oxidative stress parameters assessed. This study contributes to a better understanding of fluazuron in bovines, suggesting that the antiparasitic may affect bovine reproduction and might cause embryo loss.
Assuntos
Técnicas de Maturação in Vitro de Oócitos , Oócitos , Compostos de Fenilureia , Animais , Bovinos , Oócitos/efeitos dos fármacos , Compostos de Fenilureia/farmacologia , Técnicas de Maturação in Vitro de Oócitos/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodos , Células CHO , Cricetulus , Antiparasitários/farmacologia , Antiparasitários/toxicidade , FemininoRESUMO
Cutaneous squamous cell carcinoma (cSCC) is a common type of skin cancer that can result in significant morbidity, although it is usually well-managed and rarely metastasizes. However, the lack of commercially available cSCC cell lines hinders our understanding of this disease. This study aims to establish and characterize a new metastatic cSCC cell line derived from a Brazilian patient. A tumor biopsy was taken from a metastatic cSCC patient, immortalized, and named HCB-541 after several passages. The cytokeratin expression profile, karyotypic alterations, mutational analysis, mRNA and protein differential expression, tumorigenic capacity in xenograft models, and drug sensitivity were analyzed. The HCB-541 cell line showed a doubling time between 20 and 30 h and high tumorigenic capacity in the xenograft mouse model. The HCB-541 cell line showed hypodiploid and hypotetraploidy populations. We found pathogenic mutations in TP53 p.(Arg248Leu), HRAS (Gln61His) and TERT promoter (C228T) and high-level microsatellite instability (MSI-H) in both tumor and cell line. We observed 37 cancer-related genes differentially expressed when compared with HACAT control cells. The HCB-541 cells exhibited high phosphorylated levels of EGFR, AXL, Tie, FGFR, and ROR2, and high sensitivity to cisplatin, carboplatin, and EGFR inhibitors. Our study successfully established HCB-541, a new cSCC cell line that could be useful as a valuable biological model for understanding the biology and therapy of metastatic skin cancer.
Assuntos
Carcinoma de Células Escamosas , Mutação , Neoplasias Cutâneas , Humanos , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/patologia , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Animais , Telomerase/genética , Telomerase/metabolismo , Proteínas Proto-Oncogênicas p21(ras)/genética , CamundongosRESUMO
Zinc (Zn) plays essential roles in numerous cellular processes. However, there is limited understanding of Zn homeostasis within the bovine reproductive system. This study investigated the influence of estradiol (E2) and progesterone (P4) on Zn transporter expression and intracellular free Zn levels in bovine oviduct epithelial cells (BOEC). For this purpose, cells were harvested from slaughtered cows and cultured in vitro. Intracellular Zn concentrations were measured using FluoZin-3AM staining, while real-time polymerase chain reaction assessed Zn transporter gene expression and quantification. Overall, our results confirmed the gene expression of all the evaluated Zn transporters (ZIP6, ZIP8, ZIP14, ZnT3, ZnT7 and ZnT9), denoted and the active role of E2 and P4 in intracellular Zn regulation. Our findings suggest an interaction between Zn, E2 and P4.
Assuntos
Proteínas de Transporte , Progesterona , Zinco , Feminino , Bovinos , Animais , Progesterona/farmacologia , Progesterona/metabolismo , Zinco/farmacologia , Zinco/metabolismo , Oviductos/metabolismo , Células Epiteliais/metabolismo , Estrogênios/farmacologiaRESUMO
The discovery of antibiotics has significantly transformed the outcomes of bacterial infections in the last decades. However, the development of antibiotic resistance mechanisms has allowed an increasing number of bacterial strains to overcome the action of antibiotics, decreasing their effectiveness against infections they were developed to treat. This study aimed to evaluate the antibacterial activity of synthetic coumarins Staphylococcus aureus in vitro and analyze their interaction with the MepA efflux pump in silico. The Minimum Inhibitory Concentration (MIC) determination showed that none of the test compounds have antibacterial activity. However, all coumarin derivatives decreased the MIC of the standard efflux inhibitor ethidium bromide, indicating antibacterial synergism. On the other hand, the C14 derivative potentiated the antibacterial activity of ciprofloxacin against the resistant strain. In silico analysis showed that C9, C11, and C13 coumarins showed the most favorable interaction with the MepA efflux pump. Nevertheless, due to the present in silico and in vitro investigation limitations, further experimental research is required to confirm the therapeutic potential of these compounds in vivo.
Assuntos
Cumarínicos , Proteínas Associadas à Resistência a Múltiplos Medicamentos , Cumarínicos/farmacologia , Antibacterianos/farmacologia , Ciprofloxacina/farmacologia , Staphylococcus aureus/metabolismo , Testes de Sensibilidade Microbiana , Proteínas de Bactérias/metabolismoRESUMO
The increase in antibiotic resistance rates has attracted the interest of researchers for antibacterial compounds capable of potentiating the activity of conventional antibiotics. Coumarin derivatives have been reported to develop effective antibacterials with possible new mechanisms of action for treating infectious diseases caused by bacteria with a profile of drug resistance. In this context, the aim of the present study we have now prepared one variety of new synthetic coumarins evaluating the pharmacokinetic and chemical similarity in silico, their antimicrobial activity against Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922), and potential for the modulation of antibiotic resistance against Staphylococcus aureus (SA10) and Escherichia coli (EC06) clinical isolate bacteria by in vitro assay. The antibacterial activity and antibiotic-enhancing properties were evaluated by the broth microdilution method and pharmacokinetically characterized according to the Lipinsk rule of 5 and had their similarity analyzed in databases such as ChemBL and CAS SciFinder. The results demonstrated that only compound C13 showed significant antibacterial activity (MIC ≤256 µg/mL), and all other coumarins did not display relevant antibacterial activity (MIC ≥1024 µg/mL). However, they did modulate the antibiotics activities to norfloxacin and gentamicin, except, compound C11 to norfloxacin against Staphylococcus aureus (SA10). The in silico properties prediction and drug-likeness results demonstrated that all coumarins presented a good drug-likeness score with no violations and promising in silico pharmacokinetic profiles showing that they have the potential to be developed into an oral drug. The results indicate that the coumarin derivatives showed good in vitro antibacterial activity. These new coumarin derivatives also demonstrated the capacity to modulate antibiotic resistance with potential synergy action for current antimicrobials assayed, as antibiotic adjuvants, to reduce the emergence of antimicrobial resistance.
Assuntos
Infecções Estafilocócicas , Staphylococcus aureus , Humanos , Norfloxacino/farmacologia , Escherichia coli , Cumarínicos/farmacologia , Cumarínicos/química , Antibacterianos/farmacologia , Antibacterianos/química , Infecções Estafilocócicas/tratamento farmacológico , Bactérias , Testes de Sensibilidade MicrobianaRESUMO
Veterinary drugs are potential environmental pollutants that interfere with male reproductive function. Infertility has increased, and it is known that environmental toxins contribute to declining sperm parameters. Amitraz {N,N-[(methylamino) dimeth-ylidyne] di-2,4-xylidine} (AMZ) is a formamidine pesticide widely used as an insecticide and an acaricide. The aim of this study was to evaluate the toxicity of AMZ in bovine sperm. Three experiments using frozen-thawed bovine semen incubated with AMZ for 2 h were carried out. Negative and solvent (dimethyl sulfoxide) controls were run simultaneously with treatments. In experiment 1, the AMZ concentrations used were 10, 15 and 25 µg AMZ/ml and the sperm parameters evaluated were viability, mitochondrial activity, acrosomal status, functional membrane integrity and apoptosis. In experiments 2 and 3, 25 µg AMZ/ml was used to evaluate fertilizing capacity, embryo development and blastocyst DNA damage. In experiment 1, 25 µg AMZ/ml decreased sperm viability (P = 0.01), reduced mitochondrial activity (P = 0.03) and induced apoptosis (P < 0.01). Also, 15 and 25 µg AMZ/ml affected functional membrane integrity (P < 0.01). In experiment 2, AMZ did not alter sperm-zona binding (P = 0.40) and pronucleus formation (P = 0.36). In experiment 3, 25 µg AMZ/ml decreased the rate of embryo development (P < 0.01) and increased apoptosis (P = 0.03). These results suggest that AMZ induced alterations in bovine sperm, probably affecting male fertility at concentrations that could be present in the environment.
Assuntos
Análise do Sêmen , Preservação do Sêmen , Masculino , Animais , Bovinos , Análise do Sêmen/veterinária , Sêmen , Espermatozoides , Preservação do Sêmen/veterinária , Desenvolvimento Embrionário , Criopreservação/veterináriaRESUMO
In vitro embryo production has grown in recent decades due to its great potential for cattle production. However, the quality of in vitro-produced embryos is lower compared with those produced in vivo. The postfertilization culture environment has a major influence on bovine embryo quality. We hypothesize that the inclusion of the inclusion of alpha-lipoic acid (ALA) in the in vitro culture (IVC) medium during the first 24 h would have positive effects on embryo development in vitro and cryotolerance. The aims of this study were to evaluate the antioxidant effect of ALA in IVC medium for 24 h on bovine zygotes (21 h post in vitro fertilization, IVF), day 2 cleaved embryos (46 h post-IVF), and to assess embryo quality, developmental competence, and cryotolerance after vitrification. In all experiments, IVC medium was the Control, and 2.5 µM ALA was the treatment implemented. Viability and reactive oxygen species (ROS) levels in zygotes and day 2 embryos did not differ from the Control (P > 0.05). Supplementation with ALA increased total blastocyst and hatching rates (P < 0.05). It also improved embryo quality, evidenced by the increased blastocyst total cell number and the percentage of excellent-quality embryos observed (P < 0.05). In embryos cultured with ALA and then vitrified, ALA reduced intracellular ROS levels in warmed blastocysts (P < 0.05). In conclusion, ALA supplementation to IVC medium during 24 h is a new advantage in improving embryo quality for assisted bovine reproduction.
Assuntos
Criopreservação , Ácido Tióctico , Bovinos , Animais , Criopreservação/veterinária , Ácido Tióctico/farmacologia , Espécies Reativas de Oxigênio/farmacologia , Técnicas de Cultura Embrionária/veterinária , Vitrificação , Fertilização in vitro/veterinária , Blastocisto , Desenvolvimento EmbrionárioRESUMO
La sarcopenia está caracterizada por la baja masa y fuerza muscular en los adultos mayores. La implementación de técnicas de modificación de la conducta como la técnica de economía de fichas (TEF) puede fungir como estrategia para mejorar la adherencia al tratamiento. El objetivo fue evaluar el efecto de la TEF sobre las conductas de tratamiento: realizar la rutina de ejercicio de resistencia, consumir el suplemento de proteína y consumir el suplemento de vitamina D. Se realizó un estudio de caso de múltiples sujetos en ocho adultos mayores con sarcopenia. El estudio estuvo conformado por tres fases: pre-intervención, intervención conductual y post-intervención conductual. Se evaluó la frecuencia de las conductas de tratamiento en las tres fases, además de los cambios en masa y fuerza muscular. El análisis de los datos se realizó por medio de estadística descriptiva. Los resultados mostraron que la frecuencia de las conductas de tratamiento aumentó en el 100% de los participantes en la etapa de intervención conductual al comparar con la fase pre-intervención, valores que no regresaron a cero en la fase post-intervención conductual. El 83.3 % de los participantes dejaron de tener el diagnóstico de sarcopenia con base en los cambios de masa y fuerza muscular.
Sarcopenia is characterized by low muscle mass and strength in older adults. It is caused by decreased physical activity and quality of dietary intake. Therefore, the implementation of behavior modification techniques such as the token economy technique can serve as a strategy to improve treatment. The aim of the current study was to evaluate the effect of the token economy technique on treatment behaviors. A multiple case studies with an ABC-type design was conducted in eight older adults with sarcopenia. The intervention consisted of modification of treatment-related behaviors; in addition, changes in muscle mass and strength were assessed. The results showed that the frequency of target behaviors increased in 100% of the participants in the intervention stage compared to baseline. The frequency of the target behaviors did not return to the obtained baseline compared to the maintenance stage. 83.3% of the participants changed their sarcopenia diagnosis.
Assuntos
Humanos , Masculino , Feminino , Idoso , Idoso de 80 Anos ou mais , Reforço por Recompensa , Sarcopenia/terapia , Resistência Física , Ingestão de Alimentos , Terapia por Exercício , Cooperação e Adesão ao TratamentoRESUMO
The Barretos Cancer Hospital Animal Facility (BCHAF) is a unique facility in Brazil exclusively dedicated to working with animal models for cancer research. In this article, we briefly present our modern facility and the main experiments performed, focusing on mutant strains of mice (PTCH-knockout and ApcMin mice), xenograft models, and patient-derived xenografts (PDXs). Our results show the progress and challenges in establishing these models and the need for having an appropriate representation of our cancer population to better understand tumor biology and to identify cancer biomarkers, which could be putatively targeted, allowing for personalized therapy.
RESUMO
During healthcare catastrophes, such as the current COVID-19 pandemic, nurses are exposed to highly stressful situations derived from their work and personal activity. Development of coping strategies for such situations can improve nurses' physical and mental health. This study analyzed nurses' adaptability to change, with attention to socio-demographic variables in a COVID-19 setting, and identified the repercussions on their health. This quantitative, observational and cross-sectional study had a sample of 351 nurses aged 22 to 64 with a mean age of 40.91 (SD = 10.98). The instruments used for the study were the ADAPTA-10 questionnaire and the General Health Questionnaire (GHQ-28). It was observed that age, sex, and having a stable partner significantly influenced scores on the emotional, cognitive-behavioral, and adaptation to change factors. Finally, the emotional factor mediated between positive COVID-19 in someone close and the presence of health problems. Understanding the elements that help adapt better to change and adversity enable effective interventions to be developed for improving emotional health of nurses, especially for those in whom there are positive cases of COVID-19 in their personal or work environment.