RESUMO
BACKGROUND: Six thoracic pathologists reviewed 259 lung neuroendocrine tumours (LNETs) from the lungNENomics project, with 171 of them having associated survival data. This cohort presents a unique opportunity to assess the strengths and limitations of current World Health Organization (WHO) classification criteria and to evaluate the utility of emerging markers. PATIENTS AND METHODS: Patients were diagnosed based on the 2021 WHO criteria, with atypical carcinoids (ACs) defined by the presence of focal necrosis and/or 2-10 mitoses per 2 mm2. We investigated two markers of tumour proliferation: the Ki-67 index and phospho-histone H3 (PHH3) protein expression, quantified by pathologists and automatically via deep learning. Additionally, an unsupervised deep learning algorithm was trained to uncover previously unnoticed morphological features with diagnostic value. RESULTS: The accuracy in distinguishing typical from ACs is hampered by interobserver variability in mitotic counting and the limitations of morphological criteria in identifying aggressive cases. Our study reveals that different Ki-67 cut-offs can categorise LNETs similarly to current WHO criteria. Counting mitoses in PHH3+ areas does not improve diagnosis, while providing a similar prognostic value to the current criteria. With the advantage of being time efficient, automated assessment of these markers leads to similar conclusions. Lastly, state-of-the-art deep learning modelling does not uncover undisclosed morphological features with diagnostic value. CONCLUSIONS: This study suggests that the mitotic criteria can be complemented by manual or automated assessment of Ki-67 or PHH3 protein expression, but these markers do not significantly improve the prognostic value of the current classification, as the AC group remains highly unspecific for aggressive cases. Therefore, we may have exhausted the potential of morphological features in classifying and prognosticating LNETs. Our study suggests that it might be time to shift the research focus towards investigating molecular markers that could contribute to a more clinically relevant morpho-molecular classification.
Assuntos
Neoplasias Pulmonares , Tumores Neuroendócrinos , Humanos , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/classificação , Tumores Neuroendócrinos/patologia , Tumores Neuroendócrinos/classificação , Feminino , Antígeno Ki-67/metabolismo , Masculino , Biomarcadores Tumorais/metabolismo , Pessoa de Meia-Idade , Organização Mundial da Saúde , Histonas/metabolismo , Idoso , Prognóstico , Aprendizado ProfundoRESUMO
The Italian Association of Medical Oncology recommendations on thymic epithelial tumors, which have been drawn up for the first time in 2020 through an evidence-based approach, report indications on all the main aspects of clinical management of this group of rare diseases, from diagnosis and staging, to new available systemic treatments, such as targeted therapies and immunotherapies. A summary of key recommendations is presented here and complete recommendations are reported as Supplementary Materials, available at https://doi.org/10.1016/j.esmoop.2021.100188.
Assuntos
Neoplasias Epiteliais e Glandulares , Neoplasias do Timo , Humanos , Itália , Oncologia , Neoplasias Epiteliais e Glandulares/diagnóstico , Neoplasias Epiteliais e Glandulares/terapia , Neoplasias do Timo/diagnóstico , Neoplasias do Timo/terapiaRESUMO
Drug-eluting bioresorbable vascular scaffolds (BVSs) have emerged as a potential breakthrough for the treatment of coronary artery stenosis, providing mechanical support and drug delivery followed by complete resorption. Restenosis and thrombosis remain the primary limitations in clinical use. The study aimed to identify potential markers of restenosis and thrombosis analyzing the vascular wall cell transcriptomic profile modulation triggered by BVS at different values of shear stress (SS). Human coronary artery endothelial cells and smooth muscle cells were cultured under SS (1 and 20 dyne cm-2) for 6 h without and with application of BVS and everolimus 600 nM. Cell RNA-Seq and bioinformatics analysis identified modulated genes by direct comparison of SS conditions and Gene Ontology (GO). The results of different experimental conditions and GO analysis highlighted the modulation of specific genes as semaphorin 3E, mesenchyme homeobox 2, bone morphogenetic protein 4, (heme oxygenase 1) and selectin E, with different roles in pathological evolution of disease. Transcriptomic analysis of dynamic vascular cell cultures identifies candidate genes related to pro-restenotic and pro-thrombotic mechanisms in anin-vitrosetting of BVS, which are not adequately contrasted by everolimus addition.
Assuntos
Implantes Absorvíveis , Materiais Biocompatíveis , Biomarcadores/metabolismo , Trombose/metabolismo , Alicerces Teciduais/química , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Células Cultivadas , Reestenose Coronária/metabolismo , Vasos Coronários/citologia , Everolimo/química , Everolimo/farmacologia , Humanos , Resistência ao Cisalhamento , Transcriptoma/efeitos dos fármacosRESUMO
In addition to chronic obstructive pulmonary disease (COPD) and bronchogenic carcinoma, smoking can also cause interstitial lung diseases (ILDs) such as respiratory bronchiolitis (RB), RB with ILD (RB-ILD), desquamative interstitial pneumonia (DIP), Langerhans cell granulomatosis (LCG) and idiopathic pulmonary fibrosis-usual interstitial pneumonia (IPF-UIP). However, smoking seems to have a protective effect against hypersensitivity pneumonitis (HP), sarcoidosis and organising pneumonia (OP). High-resolution computed tomography (HRCT) has a pivotal role in the differential diagnosis. RB is extremely frequent in smokers, and is considered a marker for smoking exposure. It has no clinical relevance in itself since most patients with RB are asymptomatic. It is frequent to observe the association of RB with other smoking-related diseases, such as LCG or pulmonary neoplasms. In RB-ILD, HRCT features are more conspicuous and diffuse than in RB, but there is no definite cut-off between the two entities and any distinction can only be made by integrating imaging and clinical data. RB, RB-ILD and DIP may represent different degrees of the same pathological process, consisting in a bronchiolar and alveolar inflammatory reaction to smoking. Smoking is also a well-known risk factor for pulmonary fibrosis. Multidisciplinary discussion and follow-up can generally solve even the most difficult cases.
Assuntos
Alveolite Alérgica Extrínseca , Bronquiolite , Doenças Pulmonares Intersticiais , Alveolite Alérgica Extrínseca/diagnóstico por imagem , Alveolite Alérgica Extrínseca/etiologia , Bronquiolite/diagnóstico por imagem , Bronquiolite/etiologia , Humanos , Pulmão/diagnóstico por imagem , Doenças Pulmonares Intersticiais/diagnóstico por imagem , Doenças Pulmonares Intersticiais/etiologia , Fumar/efeitos adversos , Tomografia Computadorizada por Raios XRESUMO
The prognosis of patients affected by malignant pleural mesothelioma (MPM) is presently poor and no therapeutic strategies have improved their survival yet. Introduction of miRNA mimics to restore their reduced or absent functionality in cancer cells is considered an important opportunity and a combination of miR's might be even more effective. In the present study, miR-16 and miR-34a were transfected, singularly and in combination, in MPM cell lines H2052 and H28, and their effects on cell proliferation and sensitivity to cisplatin are reported. Interestingly, the overexpression of both miRs, alone or combined, slows down the cell cycle progression, modulates the p53 and HMGB1 expression and increases the sensitivity of cells to cisplatin, producing a marked impairment of cell proliferation and strengthening the apoptotic effect of the drug. However, the co-overexpression of the two miRs results more effective only in the regulation of the cell cycle, but does not enhance the sensitivity of MPM cells to cisplatin. Consequently, although the potential of miR-16 and miR-34a is confirmed, we must conclude that their combination does not improve the response of MPM to chemotherapy.
Assuntos
Cisplatino/farmacologia , Mesotelioma Maligno/genética , MicroRNAs/metabolismo , Neoplasias Pleurais/genética , Apoptose/genética , Ciclo Celular/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Cisplatino/uso terapêutico , Resistencia a Medicamentos Antineoplásicos/genética , Regulação Neoplásica da Expressão Gênica , Proteína HMGB1/genética , Humanos , Mesotelioma Maligno/tratamento farmacológico , Mesotelioma Maligno/patologia , MicroRNAs/genética , Neoplasias Pleurais/tratamento farmacológico , Neoplasias Pleurais/patologia , Transfecção , Proteína Supressora de Tumor p53/genéticaRESUMO
The aim of the present study was to evaluate the effects of photobiomodulation (low-level laser therapy (LLLT)) and aquatic exercise on the expression of genes related to muscle regeneration in rats. Wistar rats were divided into five groups: control group (n = 15), non-treated injury group (n = 15), injury+LLLT group (n = 15), injury+aquatic exercise group (n = 15), and injury+LLLT+aquatic exercise group (n = 15). Cryoinjury was performed on the belly of the tibialis anterior (TA) muscle. LLLT was performed daily with an AlGaAs laser (830 nm; beam spot of 0.0324 cm2, output power of 100 mW, energy density of 180 J/cm2, and 58-s exposure time). Animals were euthanized at 7, 14, and 21 days. The TA muscles were removed for gene expression analysis of TGF-ß, Myogenin, and MyoD. The results were statistically analyzed at a significance level of 5%. The cryoinjury increased the expression of genes related to muscle regeneration-MyoD, Myogenin, and TGF-ß-compared to the control group (p < 0.05); the photobiomodulation increased the expression of these genes at day 7 (p < 0.05), decreasing until day 21; and the aquatic exercise increases the expression of the three genes over time. When the two treatments were combined, the expression of the analyzed genes also increased over time. In summary, the results of our study suggest that photobiomodulation (LLLT), when applied alone in cryoinjury, is able to increase the gene expression of MyoD, Myogenin, and TGF-ß at the acute phase, while when combined with aquatic exercises, there is an increase in expression of these genes specially at the long-term treatment.
Assuntos
Terapia com Luz de Baixa Intensidade , Músculo Esquelético , Natação , Animais , Expressão Gênica , Músculo Esquelético/lesões , Ratos , Ratos WistarRESUMO
OBJECTIVE: Several lines of evidence from the last decade support the connection between nutrition and epigenetic mechanisms. In the present study we evaluated the impact of the daily dietary intake of calories and the micronutrients vitamin A, D, B1, B2, B5, C, E, copper, calcium, phosphorus, iron, iodine, selenium, manganese, potassium and sodium on the global DNA methylation profile of blood cells from older individuals. RESEARCH METHODS AND PROCEDURES: The study enrolled 126 physically independent elderly of both sexes (60 men and 66 women). For the molecular analysis, DNA samples were extracted from leukocytes and global DNA methylation was evaluated using a high throughput Elisa-based method. Correlations between global DNA methylation and the daily intake of calorie or micronutrients were evaluated using Prism5 GraphPad Software. RESULTS: A statistically significant correlation was observed between global DNA methylation and the daily caloric value (p=0.019, r=-0.21), and the intake of vitamin A (p=0.03, r=-0.18), Vitamin E (p=0.027, r=-0.20) and copper (p=0.04, r=-0.18). No correlation was observed between global DNA methylation and the daily intake of vitamin D, B1, B2, B5, C, calcium, phosphorus, iron, iodine, selenium, manganese and potassium (p>0.05). CONCLUSION: Our data demonstrate that the daily intake of calories or the micronutrients vitamin A, vitamin E and copper can potentially modulate the global DNA methylation profile of leukocytes in older adults and corroborate the notion of nutritional influences on epigenetic mechanisms.
Assuntos
Metilação de DNA/fisiologia , Dieta/normas , Ingestão de Energia/fisiologia , Leucócitos/fisiologia , Micronutrientes/uso terapêutico , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Micronutrientes/farmacologia , Pessoa de Meia-IdadeRESUMO
Fluoxetine (FLX) is an antidepressant from the selective serotonin reuptake inhibitor class that has largely been used for the treatment of depression in pregnancy. However, increasing evidences have indicated the potential of early maternal exposure to FLX to induce molecular and neuro functional effects on the offspring. In the present study we evaluated possible long lasting impacts of the maternal exposure to FLX during gestation and lactation. Female Wistar rats were gavaged with 5 mg/kg of FLX during the period that comprehends the first day of pregnancy (PD0) and the last day of lactation (LD21) (Group FLX). Control group (CTL) received a proportional volume of water. On the postnatal day 75 (PND75), male rats were euthanized and hippocampus, cortex, hypothalamus, and periaqueductal gray area (PAG) were removed. Global DNA methylation was quantified using a high-throughput ELISA-based method. In order to address neuro functional changes animals (PND75) were evaluated in the elevated plus maze and social interaction tests as well as submitted to repeated restraint stress. An increase in the global DNA methylation profile of hippocampus (p = 0.0399) was associated with the early exposure to FLX, whereas no significant change was observed in the hypothalamus (p = 0.6556), cortex (p = 0.9402) or PAG (p = 0.3822). Furthermore, early exposure to FLX was also associated with a reduction in the social interaction time (p = 0.0084) and to a decreased in the plasma corticosterone level when animals were submitted to the restraint stress (p < 0.0001). No significant change in the elevated plus maze test was associated with the early exposure to FLX. In summary, our data demonstrate that maternal exposure to FLX during gestation and lactation results in a long lasting impact on the DNA methylation of hippocampus, and affects the social behavior and the corticosterone response to stress.
Assuntos
Encéfalo/efeitos dos fármacos , Encéfalo/crescimento & desenvolvimento , Metilação de DNA/efeitos dos fármacos , Fluoxetina/efeitos adversos , Efeitos Tardios da Exposição Pré-Natal , Comportamento Social , Animais , Animais Recém-Nascidos , Comportamento Animal/efeitos dos fármacos , Epigênese Genética/efeitos dos fármacos , Feminino , Lactação , Masculino , Exposição Materna , Gravidez , Ratos Wistar , Estresse Psicológico/metabolismoRESUMO
Extracellular matrix (ECM) is a fundamental component of multicellular organisms. Alteration of its structure and/or molecular composition are associated with several pathologies, among the others with atherosclerosis. To determine how the overall ECM architecture of a tissue as complex as the atheroma may change under varying pathological conditions constitutes a great technical challenge. This entails removing cell components and solubilisation of fibrillar proteins in order to allow enzymatic digestion and mass spectrometry analyses. This work aimed at testing and assessing an easy to use, standardized and reproducible proteomics protocol to map ECM proteins in carotid plaque specimens. To this end, plaques from endarterectomies were incubated in different buffers and the resulting solutions and tissue homogenates after incubation were processed for mass spectrometry. Comparison of the different workflows showed that 4M Guanidine treatment (following 0.5M NaCl and 0.08% SDS incubations) is the most promising in terms of ECM enrichment. The protocol can also be used to identify different and complementary classes of proteins both in plaque extracts and homogenates.
Assuntos
Endarterectomia das Carótidas , Matriz Extracelular/metabolismo , Placa Aterosclerótica/patologia , Placa Aterosclerótica/cirurgia , Proteômica , Feminino , Humanos , Masculino , Placa Aterosclerótica/metabolismoRESUMO
The potential of behavioral stress to affect epigenetic mechanisms of non-encephalic tissues is still underestimated. In the present study we evaluated the effects of chronic behavioral stress on the DNA methylation profile of rat lung cells. Furthermore, we evaluated the potential of physical exercise to modulate the changes evoked by behavioral stress in lung cells. Male Wistar rats were divided into four experimental groups: (1) animals submitted to chronic restraint stress (CRS) (ST group) during the period of the 67th-80th postnatal day (PND); (2) animals submitted to physical exercise (EX group) during the 53rd-79th PND; (3) animals submitted to swimming during the 53rd-79th PND and to CRS during the 67th-80th PND (EX-ST group); and (4) animals not submitted to stress or swimming protocols (CTL). Global DNA methylation was quantified using an ELISA-based approach and gene expression was evaluated by real time PCR. A decreased global DNA methylation profile was observed in the ST group, however physical exercise demonstrated protection of lung cells from this stress-related hypomethylation. Increased expression of the Dnmt1 gene was evidenced in the ST group, whereas physical exercise was shown to protect lung cells from this stress-related effect in the EX-ST group. Comparative analysis of the ST and EX groups revealed opposite effects on the expression of Dnmt3a and Dnmt3b; however, a stress-related increase in expression of Dnmt3a and Dnmt3b was not seen in the EX-ST group. Our data showed that behavioral stress induced significant changes in the DNA methylation profile of rat lung cells and that this could be modulated by physical exercise.
Assuntos
Comportamento Animal/fisiologia , Metilação de DNA , Restrição Física , Animais , Epigênese Genética/genética , Masculino , Condicionamento Físico Animal , Ratos Wistar , Restrição Física/métodos , Natação/fisiologiaRESUMO
Epigenetics has recently been linked to molecular adaptive responses evoked by physical exercise and stress. Herein we evaluated the effects of physical exercise on global DNA methylation and expression of the Dnmt1 gene in the rat brain and also verified its potential to modulate responses evoked by repeated restraint stress (RRS). Wistar rats were classified into the following experimental groups: (1) physically active (EX): animals submitted to swimming during postnatal days 53-78 (PND); (2) stress (ST): animals submitted to RRS during 75-79PND; (3) exercise-stress (EX-ST): animals submitted to swimming during 53-78PND and to RRS during 75-79PND, and (4) control (CTL): animals that were not submitted to intervention. Samples from the hippocampus, cortex and hypothalamus were obtained at 79PND. The global DNA methylation profile was assessed using an ELISA-based method and the expression of Dnmt1 was evaluated by real-time PCR. Significantly increased methylation was observed in the hypothalamus of animals from the EX group in comparison to CTL. Comparative analysis involving the EX-ST and ST groups revealed increased global DNA methylation in the hippocampus, cortex, and hypothalamus of EX-ST, indicating the potential of physical exercise in modulating the responses evoked by RRS. Furthermore, decreased expression of the Dnmt1 gene was observed in the hippocampus and hypothalamus of animals from the EX-ST group. In summary, our data indicate that physical exercise affects DNA methylation of the hypothalamus and might modulate epigenetic responses evoked by RRS in the hippocampus, cortex, and hypothalamus.
Assuntos
Adaptação Fisiológica/fisiologia , Córtex Cerebral/metabolismo , DNA (Citosina-5-)-Metiltransferases/metabolismo , Epigênese Genética , Hipocampo/metabolismo , Hipotálamo/metabolismo , Condicionamento Físico Animal/fisiologia , Estresse Psicológico/metabolismo , Animais , DNA (Citosina-5-)-Metiltransferase 1 , Metilação de DNA/fisiologia , Masculino , Ratos , Ratos Wistar , Restrição FísicaRESUMO
A high-fat diet is known to induce atherosclerosis in animal models. Dietary factors and timing of atherogenic food delivery may affect plasma lipoprotein content composition and its potential atherogenic effect. Increasingly often, humans spend periods/days eating in a completely unregulated way, ingesting excessive amounts of food rich in oils and fats, alternating with periods/days when food intake is more or less correct. We investigate the effect on lipid homeostasis of a high-fat diet administered either continuously or intermittently. We investigated control pigs receiving standard diet (C, n=7), pigs receiving a high-fat diet every day for 10 weeks (CHF, n=5), and pigs receiving a high-fat diet every other week for 10 weeks (IHF, n=7). IHF animals were shown to have a different lipid profile compared with CHF animals, with a significant increase in high-density lipoproteins (HDL) levels with respect to C and CHF groups. CHF also showed significantly higher values of TC/HDL cholesterol compared with C and IHF. Hepatic expression analysis of genes involved in lipid homeostasis showed an increasing trend of nuclear receptor LXRα along with its target genes in the CHF group and in the IHF group, whereas SREBP2 and LDLr were significantly inhibited. A significant correlation was found between ABCA1 expression and circulating levels of HDL-C. Periodic withdrawals of a high-fat atherogenic diet compared with a regular administration results in a different adaptive response of lipoprotein metabolism, which leads to a significantly higher plasma level of HDL-C and lower TC/HDL-C.
Assuntos
Dieta Aterogênica/veterinária , Metabolismo dos Lipídeos , Lipídeos/sangue , Suínos/metabolismo , Adaptação Fisiológica , Animais , MasculinoRESUMO
BACKGROUND: Pulmonary carcinoids (PCs) are rare tumors. As there is a paucity of randomized studies, this expert consensus document represents an initiative by the European Neuroendocrine Tumor Society to provide guidance on their management. PATIENTS AND METHODS: Bibliographical searches were carried out in PubMed for the terms 'pulmonary neuroendocrine tumors', 'bronchial neuroendocrine tumors', 'bronchial carcinoid tumors', 'pulmonary carcinoid', 'pulmonary typical/atypical carcinoid', and 'pulmonary carcinoid and diagnosis/treatment/epidemiology/prognosis'. A systematic review of the relevant literature was carried out, followed by expert review. RESULTS: PCs are well-differentiated neuroendocrine tumors and include low- and intermediate-grade malignant tumors, i.e. typical (TC) and atypical carcinoid (AC), respectively. Contrast CT scan is the diagnostic gold standard for PCs, but pathology examination is mandatory for their correct classification. Somatostatin receptor imaging may visualize nearly 80% of the primary tumors and is most sensitive for metastatic disease. Plasma chromogranin A can be increased in PCs. Surgery is the treatment of choice for PCs with the aim of removing the tumor and preserving as much lung tissue as possible. Resection of metastases should be considered whenever possible with curative intent. Somatostatin analogs are the first-line treatment of carcinoid syndrome and may be considered as first-line systemic antiproliferative treatment in unresectable PCs, particularly of low-grade TC and AC. Locoregional or radiotargeted therapies should be considered for metastatic disease. Systemic chemotherapy is used for progressive PCs, although cytotoxic regimens have demonstrated limited effects with etoposide and platinum combination the most commonly used, however, temozolomide has shown most clinical benefit. CONCLUSIONS: PCs are complex tumors which require a multidisciplinary approach and long-term follow-up.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Tumor Carcinoide/terapia , Neoplasias Pulmonares/terapia , Broncoscopia , Carboplatina/administração & dosagem , Doença Cardíaca Carcinoide/diagnóstico por imagem , Tumor Carcinoide/diagnóstico , Cisplatino/administração & dosagem , Dacarbazina/administração & dosagem , Dacarbazina/análogos & derivados , Etoposídeo/administração & dosagem , Europa (Continente) , Humanos , Neoplasias Pulmonares/diagnóstico , Pneumonectomia , Tomografia por Emissão de Pósitrons , Receptores de Somatostatina/metabolismo , Sociedades Médicas , Temozolomida , Tomografia Computadorizada por Raios X , UltrassonografiaRESUMO
Thymic epithelial tumors (TETs) are rare primary mediastinal tumors arising from thymic epithelium. Their rarity and complexity hinder investigations of their causes and therapy development. Here, we summarize the existing knowledge regarding medical treatment of these tumors, and thoroughly review the known genetic aberrations associated with TETs and the present status of potential biological treatments. Epidermal growth factor receptor (EGFR), stem-cell factor receptor, insulin-like growth factor-1 receptor (IGF1R), and vascular endothelial growth factors (VEGF-A, VEGF-B, and VEGF-2) are overexpressed in TETs. EGFR overexpression in TETs is associated with higher stage, and IGF1R overexpression has poor prognostic value. Data indicate that anti-IGF1R monoclonal antibodies, and inhibitors of angiogenesis, somatostatin receptors, histone deacetylase, mammalian target of rapamycin, and cyclin-dependent kinases may be active against TETs. Continued investigations in this field could lead to advancement of targeted and biological therapies for TETs.
Assuntos
Antineoplásicos/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Produtos Biológicos/uso terapêutico , Biomarcadores Tumorais/antagonistas & inibidores , Terapia de Alvo Molecular , Neoplasias Epiteliais e Glandulares/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Neoplasias do Timo/tratamento farmacológico , Animais , Antineoplásicos/efeitos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Produtos Biológicos/efeitos adversos , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Humanos , Terapia de Alvo Molecular/efeitos adversos , Estadiamento de Neoplasias , Neoplasias Epiteliais e Glandulares/genética , Neoplasias Epiteliais e Glandulares/metabolismo , Neoplasias Epiteliais e Glandulares/patologia , Neoplasias do Timo/genética , Neoplasias do Timo/metabolismo , Neoplasias do Timo/patologia , Resultado do TratamentoRESUMO
Fluoxetine is an antidepressant that has been largely used for treatment of depression in pregnancy. In the present study we evaluated the effects of the exposure to fluoxetine during gestation and lactation on DNA methylation of rat brain regions. Female Wistar rats were treated with 5mg/kg of fluoxetine during pregnancy and lactation. In order to assess the effects of fluoxetine in the context of maternal folic acid supplementation we performed an additional combined treatment composed by folic acid (8 mg/kg/day) and fluoxetine (5 mg/kg/day). On the postnatal day 22, male rats were euthanized and hippocampus, cortex, hypothalamus, and periaqueductal gray area were removed. Global DNA methylation was quantified using a high-throughput ELISA-based method. Neurofunctional changes were addressed using validated behavioral tests: hot plate, elevated plus maze and open field. A decrease in the global DNA methylation profile of hippocampus was associated to the exposure to fluoxetine, whereas an increase in methylation was observed in cortex. The combined treatment induced an increase in the methylation of hippocampus indicating the potential of folic acid to modulate this epigenetic alteration. Increase in the latency to the thermal nociceptive response was observed in animals exposed to fluoxetine whereas this effect was abolished in animals from the combined treatment. In summary we demonstrated that exposure to fluoxetine during gestation and lactation affect the DNA methylation of brain and the nociceptive response of rats. Furthermore our data reveal the potential of folic acid to modulate epigenetic and functional changes induced by early exposure to fluoxetine.
Assuntos
Antidepressivos de Segunda Geração/toxicidade , Metilação de DNA/efeitos dos fármacos , Fluoxetina/toxicidade , Ácido Fólico/farmacologia , Lactação/efeitos dos fármacos , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Complexo Vitamínico B/farmacologia , Análise de Variância , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Comportamento Exploratório/efeitos dos fármacos , Comportamento Exploratório/fisiologia , Feminino , Idade Gestacional , Hiperalgesia/etiologia , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Aprendizagem em Labirinto/fisiologia , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Efeitos Tardios da Exposição Pré-Natal/fisiopatologia , Ratos , Ratos WistarRESUMO
Lung neuroendocrine tumors are catalogued in four categories by the World Health Organization (WHO 2004) classification. Its reproducibility and prognostic efficacy was disputed. The WHO 2010 classification of digestive neuroendocrine neoplasms is based on Ki67 proliferation assessment and proved prognostically effective. This study aims at comparing these two classifications and at defining a prognostic grading system for lung neuroendocrine tumors. The study included 399 patients who underwent surgery and with at least 1 year follow-up between 1989 and 2011. Data on 21 variables were collected, and performance of grading systems and their components was compared by Cox regression and multivariable analyses. All statistical tests were two-sided. At Cox analysis, WHO 2004 stratified patients into three major groups with statistically significant survival difference (typical carcinoid vs atypical carcinoid (AC), P=0.021; AC vs large-cell/small-cell lung neuroendocrine carcinomas, P<0.001). Optimal discrimination in three groups was observed by Ki67% (Ki67% cutoffs: G1 <4, G2 4-<25, G3 ≥25; G1 vs G2, P=0.021; and G2 vs G3, P≤0.001), mitotic count (G1 ≤2, G2 >2-47, G3 >47; G1 vs G2, P≤0.001; and G2 vs G3, P≤0.001), and presence of necrosis (G1 absent, G2 <10% of sample, G3 >10% of sample; G1 vs G2, P≤0.001; and G2 vs G3, P≤0.001) at uni and multivariable analyses. The combination of these three variables resulted in a simple and effective grading system. A three-tiers grading system based on Ki67 index, mitotic count, and necrosis with cutoffs specifically generated for lung neuroendocrine tumors is prognostically effective and accurate.
Assuntos
Carcinoma Neuroendócrino/patologia , Neoplasias Pulmonares/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/análise , Carcinoma Neuroendócrino/classificação , Carcinoma Neuroendócrino/mortalidade , Criança , Estudos de Coortes , Estudos Transversais , Medicina Baseada em Evidências , Feminino , Humanos , Itália/epidemiologia , Estimativa de Kaplan-Meier , Antígeno Ki-67/análise , Estudos Longitudinais , Neoplasias Pulmonares/classificação , Neoplasias Pulmonares/mortalidade , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Organização Mundial da Saúde , Adulto JovemRESUMO
This study aimed at challenging pulmonary large cell carcinoma (LLC) as tumor entity and defining different subgroups according to immunohistochemical and molecular features. Expression of markers specific for glandular (TTF-1, napsin A, cytokeratin 7), squamous cell (p40, p63, cytokeratins 5/6, desmocollin-3), and neuroendocrine (chromogranin, synaptophysin, CD56) differentiation was studied in 121 LCC across their entire histological spectrum also using direct sequencing for epidermal growth factor receptor (EGFR) and v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations and FISH analysis for ALK gene translocation. Survival was not investigated. All 47 large cell neuroendocrine carcinomas demonstrated a true neuroendocrine cell lineage, whereas all 24 basaloid and both 2 lymphoepithelioma-like carcinomas showed squamous cell markers. Eighteen out of 22 clear cell carcinomas had glandular differentiation, with KRAS mutations being present in 39 % of cases, whereas squamous cell differentiation was present in four cases. Eighteen out of 20 large cell carcinomas, not otherwise specified, had glandular differentiation upon immunohistochemistry, with an exon 21 L858R EGFR mutation in one (5 %) tumor, an exon 2 KRAS mutation in eight (40 %) tumors, and an ALK translocation in one (5 %) tumor, whereas two tumors positive for CK7 and CK5/6 and negative for all other markers were considered adenocarcinoma. All six LCC of rhabdoid type expressed TTF-1 and/or CK7, three of which also harbored KRAS mutations. When positive and negative immunohistochemical staining for these markers was combined, three subsets of LCC emerged exhibiting glandular, squamous, and neuroendocrine differentiation. Molecular alterations were restricted to tumors classified as adenocarcinoma. Stratifying LCC into specific categories using immunohistochemistry and molecular analysis may significantly impact on the choice of therapy.
Assuntos
Carcinoma de Células Grandes/classificação , Neoplasias Pulmonares/classificação , Adulto , Idoso , Quinase do Linfoma Anaplásico , Carcinoma de Células Grandes/genética , Carcinoma de Células Grandes/patologia , Feminino , Humanos , Imuno-Histoquímica , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Biologia Molecular , Mutação , Estadiamento de Neoplasias , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas p21(ras) , Receptores Proteína Tirosina Quinases/genética , Proteínas ras/genéticaRESUMO
Angiotensins (Angs) modulate blood pressure, hydro-electrolyte composition, and antinociception. Although Ang (5-8) has generally been considered to be inactive, we show here that Ang (5-8) was the smallest Ang to elicit dose-dependent responses and receptor-mediated antinociception in the rat ventrolateral periaqueductal gray matter (vlPAG). Ang (5-8) antinociception seems to be selective, because it did not alter blood pressure or act on vascular or intestinal smooth muscle cells. The non-selective Ang-receptor (Ang-R) antagonist saralasin blocked Ang (5-8) antinociception, but selective antagonists of Ang-R types I, II, IV, and Mas did not, suggesting that Ang (5-8) may act via an unknown receptor. Endopeptidase EP 24.11 and amastatin-sensitive aminopeptidase from the vlPAG catalyzed the synthesis (from Ang II or Ang III) and inactivation of Ang (5-8), respectively. Selective inhibitors of neuronal-nitric oxide (NO) synthase, soluble guanylyl cyclase (sGC) and a non-selective opioid receptor (opioid-R) inhibitor blocked Ang (5-8)-induced antinociception. In conclusion, Ang (5-8) is a new member of the Ang family that selectively and strongly modulates antinociception via NO-sGC and endogenous opioid in the vlPAG.
Assuntos
Angiotensina I/farmacologia , Guanilato Ciclase/metabolismo , Óxido Nítrico/metabolismo , Nociceptividade/efeitos dos fármacos , Peptídeos Opioides/metabolismo , Fragmentos de Peptídeos/farmacologia , Substância Cinzenta Periaquedutal/efeitos dos fármacos , Receptores Citoplasmáticos e Nucleares/metabolismo , Transdução de Sinais/efeitos dos fármacos , Antagonistas de Receptores de Angiotensina/farmacologia , Animais , Aorta/efeitos dos fármacos , Relação Dose-Resposta a Droga , Frequência Cardíaca/fisiologia , Masculino , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Peptídeos Opioides/antagonistas & inibidores , Ratos , Ratos Wistar , Saralasina/farmacologia , Guanilil Ciclase Solúvel , Teprotida/farmacologiaRESUMO
Sudden death in a 66-year-old woman with squamous cell carcinoma of the oral cavity and exclusive metastatic involvement of the right stellate ganglion and right nerve vagus is reported. The patient also suffered from paroxysmal atrial fibrillation treated with quinidine. An autopsy showed exclusive metastases to the right stellate ganglion and vagus nerve, along with decreased nerve fibre density in the ventricular myocardium suggesting that Wallerian axon degeneration of cardiac fibres was responsible for sudden death.
Assuntos
Carcinoma de Células Escamosas/complicações , Carcinoma de Células Escamosas/secundário , Neoplasias dos Nervos Cranianos/complicações , Neoplasias dos Nervos Cranianos/secundário , Morte Súbita/etiologia , Neoplasias Bucais/patologia , Neoplasias do Sistema Nervoso Periférico/complicações , Neoplasias do Sistema Nervoso Periférico/secundário , Gânglio Estrelado , Nervo Vago , Idoso , Feminino , HumanosRESUMO
BACKGROUND: Renal cell carcinomas (RCCs) are rarely described in transplanted kidneys. Available therapeutic strategies range from allograft nephrectomy to nephron-sparing procedures such as partial nephrectomy or image-guided thermal ablation. Percutaneous radiofrequency ablation (RFA) is a minimally invasive technique which provides promising oncologic outcomes in small allograft RCCs while preserving allograft function. So far, only a few cases have been reported in the transplant setting. We describe a renal transplant RCC successfully approached by ultrasound-guided RFA. METHODS: A 42-year-old renal transplant recipient developed a small subcapsular allograft RCC at 11 years after transplantation. The decline in glomerular filtration rare prompted us to preserve as much parenchyma as possible. Ultrasound-guided RFA was performed under light sedation and local analgesia in a single session with a Starbust Talon needle. RESULTS: Postablation contrast-enhanced ultrasound displayed a 25×23 mm avascular area of complete necrosis. After 3 months gadolinium-enhanced magnetic resonance imaging confirmed the absence of viable tumor tissue and while the patient did not experience any graft function reduction (serum creatinine 2.6 mg/dL). CONCLUSIONS: Image-guided RFA represents a promising therapeutic modality for small allograft RCCs in recipients with mild graft dysfunction and/or elevated surgical risk. It is associated with low morbidity and parenchymal preservation.