RESUMO
Zebrafish is a popular toxicology model and provides an ethically acceptable small-scale analysis system with the complexity of a complete organism. Our goal is to further validate this model for its regulatory use for reproductive and developmental defects by testing the compounds indicated in the "Guideline on detection of reproductive and developmental toxicity for human pharmaceuticals" (ICH S5(R3) guideline.) To determine the embryotoxic and developmental risk of the 32 reference compounds listed in the ICH S5(R3) guideline, the presence of morphological alterations in zebrafish embryos was analyzed at two different stages to calculateLC50 and EC50 values for each stage. Teratogenic Indexes were established as the ratio between LC50 and EC50 critical for the proper compound classification as teratogenic when it is ≥ 2. A total of three biological replicates have been conducted to study the reproducibility of the assay. The chemicals' concentration in the medium and internally in the zebrafish embryos was evaluated. In this study, the 3 negative compounds were properly categorized while 23 compounds out of the 29 reference ones (sensitivity of 79.31%) were classified as teratogenic in zebrafish. The 6 that had false-negative results were classified 4 as inconclusive, 1 as not toxic, and 1 compound resulted toxic for zebrafish embryos under testing conditions. After the bioavailability experiments, some of the obtained inconclusive results were refined. The developmental defects assay in zebrafish gives an accuracy of 89.66%, sensitivity of 88.46%, specificity and repeatability of 100% compared to mammals; therefore, this is a well-integrated strategy using New Alternative Methods, to minimize the use of animals in developmental toxicity studies.
Assuntos
Teratogênese , Peixe-Zebra , Animais , Humanos , Reprodutibilidade dos Testes , Embrião não Mamífero , Teratogênicos/toxicidade , MamíferosRESUMO
Thyroid disruption is an increasingly recognized issue in the use and development of chemicals and new drugs, especially to help toxicologist to complement the reproductive and developmental toxicology information of chemicals. Still, adequate assessment methods are scarce and often suffer a trade-off between physiological relevance and labor- and cost-intensive assays. Here, we present a tiered approach for a medium-throughput screening of chemicals to identify their thyroid disrupting potential in zebrafish embryos as a New Approach Methodology (NAM). After identifying the maximum tolerated concentrations, we exposed zebrafish larvae to sub-adverse effect levels of the reference compounds benzophenone-2, bisphenol A, phenylthiourea, potassium perchlorate, propylthiouracil, and phloroglucinol to exclude any systemic toxicity. Applying the transgenic zebrafish line that carries a gene for the red fluorescence protein (Tg(tg:mCherry)) under the thyroglobulin promoter, we could identify the thyroid disrupting effects of the chemicals by a time and cost-effective image analysis measuring the fluorescence levels in the thyroid glands. Our observations could be confirmed by altered expression patterns of genes involved in the hypothalamus-pituitary-thyroid (HPT) axis. Finally, to anchor the observed thyroid disruption, we determined some changes in the Thyroid hormone levels of triiodothyronine (T3) and Thyroxine (T4) using a newly developed liquid chromatography mass spectrometric (LCMS) method. The presented approach carries the potential to extend the toolbox for legislative authorities and chemical producers for the assessment of thyroid-specific endocrine disruption and to overcome current challenges in the evaluation of endocrine disruptors.
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INTRODUCTION: Lung transplantation is the final treatment option in patients with respiratory failure. Morbidity and mortality rates associated with the management of complications is high despite advances. Postoperative complications include acute transplant rejection, bronchiolitis obliterans, and infections. Because of that, the success of this treatment option depends on the correct choice of donor and candidates to receive a transplant. OBJECTIVE: This study aims to perform a survival analysis of transplanted patients in our center and determine predictive variables of mortality. PATIENTS AND METHODS: This study is a retrospective assessment of data collected from 510 patients at the Hospital University Reina Sofía from October 1993 to December 31, 2016. Patients who were retransplanted were excluded. We collected data regarding basal characteristics of the donors and candidates to receive a transplant. We analyzed the impact in terms of future survival of basal variables from donor and donor recipients. RESULTS: Five hundred ten patients were included (average age 44 ± 17 years, 69% male), with a maximum follow-up period of 21.6 years (average follow-up 4.52 years, interquartile ratio: 0.13 to 6.97 years). Two hundred twenty-seven patients died (54.3% of the total amount). The influence of donor's basal characteristics on mortality was analyzed; moreover, the relationship between basal variables and survival were analyzed using univariate analysis. Available variables were analyzed through multivariate analysis. CONCLUSION: Lung transplantation is a treatment option with an acceptable risk of morbidity and mortality. Increased awareness of features of evolution could help to reduce postoperative complications.
Assuntos
Transplante de Pulmão/mortalidade , Complicações Pós-Operatórias/epidemiologia , Adolescente , Adulto , Feminino , Rejeição de Enxerto/epidemiologia , Humanos , Estimativa de Kaplan-Meier , Transplante de Pulmão/efeitos adversos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Estudos Retrospectivos , Análise de SobrevidaRESUMO
OBJECTIVE: To investigate the capacity of monocytes from women with endometriosis to influence endometrial cell proliferation. DESIGN: Uterine endometrial cells were cultured in the presence and absence of autologous blood monocytes for 72 hours before assessment of endometrial cell proliferation by thymidine incorporation. SETTING: Patients were tested at initial presentation for evaluation of infertility and/or endometriosis. PATIENTS, PARTICIPANTS: Fertile controls, n = 17; infertile controls, n = 9; untreated endometriosis, n = 29. INTERVENTIONS: None. RESULTS: Endometrial cell proliferation was enhanced significantly by blood monocytes in patients with endometriosis but was suppressed significantly by blood monocytes in fertile controls. Endometrial cell proliferation was not affected significantly by blood monocytes in infertile controls analyzed as a group, but a subset of infertile patients also showed enhancement of endometrial cell proliferation by blood monocytes. CONCLUSIONS: Blood monocytes from patients with endometriosis and a subset of patients with unexplained infertility enhance autologous endometrial cell proliferation, whereas blood monocytes from fertile patients suppress endometrial cell proliferation. The capacity of monocytes to enhance endometrial cell proliferation appears to require both monocyte-derived factors that stimulate endometrial cell proliferation and endometrial cells capable of responding to those stimulatory factors. If either of these factors is absent, monocytes either suppress or have no effect on endometrial cell proliferation.