Expresión de los receptores de estrógenos y de progesterona en tejido mamario normal y tumoral canino

Los objetivos de esta tesis fueron estudiar si la distribución, concentración y afinidad de los receptores de estrógenos (RE) y progesterona (RP) en la glándula mamaria están vinculados a la concentración de Progesterona (P4) sérica en diferentes etapas del ciclo en perras normales y en perras con tumores mamarios. Se realizó un diseño en perras normales y otro en perras con tumores mamarios. Se utilizaron 12 perras adultas sanas enteras y 4 prepúberes y 22 adultas (15 enteras, 7 castradas) con tumores mamarios. Se obtuvieron muestras de tejido mamario durante las cirugías. Se determinó P4 por RIA, la distribución de RE? y RP por inmunohistoquímica y la concentración de RE y RP por ensayos de unión. Los resultados fueron analizados por ANOVA, expresados como la media ± EEM, considerando significativo P<0,05. La concentración (ng/mL) de P4 sérica fue mayor en diestro que en proestro, estro y anestro en ambos diseños. Se detectó inmunomarcado a RE? y RP en tejido conectivo, epitelios alveolar y de conductos en perras adultas normales y con tumores mamarios sugiriendo que los tres compartimientos son blanco de acción de los estrógenos y P4. La intensidad de inmunomarcado a RE? y RP fue mayor en epitelio alveolar que en tejido conectivo. La intensidad de inmunomarcado a RE? en epitelio de conductos fue mayor en anestro que en proestro, estro y diestro y la de RP en tejido conectivo fue mayor al estro que en proestro, diestro y anestro. La afinidad de ambos receptores fue similar en ambos diseños. La concentración (fmol/prot) de RE y RP fue menor en perras en diestro que en proestro y estro. El perfil de distribución y concentración de RE y RP en nuestro trabajo está vinculado a los niveles circulantes de P4, en acuerdo con lo reportado para RE pero no para RP. En perras prepúberes la concentración de ambos receptores fue menor que en adultas, en acuerdo con el escaso desarrollo del tejido mamario. En perras enteras con tumores mamarios la intensidad de inmunomarcado a RE? fue mayor en tumores benignos que malignos e hiperplasias y fue mayor en tejido conectivo que en ambos epitelios. En tumores benignos también fue mayor en tejido conectivo que en ambos epitelios. La de RP en epitelios de conductos tendió a ser mayor en hiperplasias que en tumores malignos. En perras castradas no hubo diferencias en los parametros estudiados. La concentración de RE fue mayor en tumores benignos que malignos e hiperplasias y la de RP fue similar en los diferentes tipos de tumores. Por ensayos de unión todas las muestras de tumores fueron RE y RP positivos, sugiriendo que las vías relacionadas con los estrógenos y progestinas pueden proporcionar dianas eficaces para el tratamiento del cáncer de mama

Structural validation of a realistic wing structure: the RIBES test article

Several experimental test cases are available in literature to study and validate fluid structure interaction methods. They, however, focus the attention mainly on replicating typical cruising aerodynamic conditions forcing the adoption of fully steel made models able to operate with the high loads generated in high speed facilities. This translates in a complete loss of similitude with typical realistic aeronautical wing structures configurations. To reverse this trend, and to better study the aerolastic mechanism from a structural point of view, an aeroelastic measurement campaign was carried within the EU RIBES project. A half wing model for wind tunnel tests was designed and manufactured replicating a typical metallic wing box structure, producing a database of loads, pressure, stress and deformation measurements. In this paper the design, manufacturing and validation activities performed within the RIBES project are described, with a focus on the structural behavior of the test article. All experimental data and numerical models are made freely available to the scientific community

Comparação de técnicas de avaliação do colágeno em secções histológicas de ratos submetidos a radioterapia

Introdução: o colágeno é uma proteína fibrosa que participa de diversos processos fisiológicos e patológicos. Dada sua importância, a avaliação desse componente tem sido de grande relevância em variadas pesquisas biológicas. Objetivo: verificar a concordância entre métodos quantitativos na análise do colágeno em secções teciduais histológicas. Metodologia: trata-se de estudo experimental, que utilizou secções teciduais coradas em Sirius Vermelho de 28 ratos Wistar, pertencentes a dois grupos experimentais, com animais submetidos à radioterapia, sem G1 ou com G2, e com úlcera produzida em dorso lingual para avaliação do reparo tecidual. Foram estabelecidos dois métodos para comparação das técnicas de avaliação do colágeno: morfometria por contagem manual de pontos com duas grades (320 e 588 pontos), e morfometria por segmentação de cor. Foram utilizados testes não paramétricos Mann Whitney, para as comparações entre os grupos, e Friedman e Nemenyi, para as comparações entre as técnicas. O teste de correlação de Pearson foi usado para avaliar a quantidade de colágeno entre os métodos. O nível de significância foi de 5%. Resultados: em ambos os grupos experimentais houve correlação significativa entre a quantidade de colágeno avaliada pela técnica de 320 e 588 pontos. Nos dois grupos, a porcentagem de colágeno foi significativamente maior quando avaliada pela técnica de 588 pontos do que pela técnica por segmentação por cor (p&lt;0,05). No grupo G2, houve correlação significativa entre a quantidade de colágeno avaliada pela técnica de 588 pontos e por segmentação por cor (p&lt;0,05). Pelas técnicas de 588 pontos e segmentação por cor, a quantidade de colágeno foi significativamente maior no grupo com lesão (p&lt;0,05). Conclusão: ainda que alguns resultados tenham se mostrado semelhantes entre as técnicas de contagem de pontos com grade de 588 pontos e segmentação por cor, os dados sugerem que a técnica de 588 pontos é mais eficaz na quantificação do colágeno

A djuvant treatment in patients at high risk of recurrence of thymoma: Efficacy and safety of a three-dimensional conformal radiation therapy regimen

The clinical benefits of postoperative radiation therapy (PORT) for patients with thymoma are still controversial. In the absence of defined guidelines, prognostic factors such as stage, status of surgical margins, and histology are often considered to guide the choice of adjuvant treatment (radiotherapy and/or chemotherapy). In this study, we describe our single-institution experience of three-dimensional conformal PORT administered as adjuvant treatment to patients with thymoma. METHODS: Twenty-two consecutive thymoma patients (eleven male and eleven female) with a median age of 52 years and treated at our institution by PORT were analyzed. The patients were considered at high risk of recurrence, having at least one of the following features: stage IIB or III, involved resection margins, or thymic carcinoma histology. Three-dimensional conformal PORT with a median total dose on clinical target volume of 50 (range 44-60) Gy was delivered to the tumor bed by 6-20 MV X-ray of the linear accelerator. Follow-up after radiotherapy was done by computed tomography scan every 6 months for 2 years and yearly thereafter. RESULTS: Two of the 22 patients developed local recurrence and four developed distant metastases. Median overall survival was 100 months, and the 3-year and 5-year survival rates were 83% and 74%, respectively. Median disease-free survival was 90 months, and the 5-year recurrence rate was 32%. On univariate analysis, pathologic stage III and presence of positive surgical margins had a significant impact on patient prognosis. Radiation toxicity was mild in most patients and no severe toxicity was registered. CONCLUSION: Adjuvant radiotherapy achieved good local control and showed an acceptable toxicity profile in patients with high-risk thymoma

CARACTERIZAÇÃO HISTOLÓGICA DA MUCOSA ORAL NORMAL DE RATOS WISTAR: ESTUDO COMPARATIVO COM MUCOSA ORAL DE HUMANOS

Objetivo: Descrever os aspectos microscópicos de normalidade da mucosa jugal e língua de ratos Wistar e comparar com as características descritas na literatura destas regiões em humanos. Materiais e métodos: Estudo observacional descritivo, que utilizou amostras da mucosa jugal e da língua de vinte ratos. Com secções coradas em HE e Sirius red, realizou-se análise morfológica dos tecidos epitelial e conjuntivo, e análise descritiva do padrão de deposição colagênico, respectivamente. Resultados: Na mucosa jugal, identificou-se tecido epitelial de revestimento pavimentoso estratificado ortoqueratinizado, de espessura delgada. Presença das camadas basal, espinhosa, granulosa e córnea. Houve presença de cristas epiteliais na interface epitélio-conjuntivo. No tecido conjuntivo, observou-se expressiva concentração de fibras colagênicas e contingente significativo de fibroblastos, além de vasos sanguíneos de pequeno calibre e discreto infiltrado inflamatório residente. A mucosa lingual apresentou epitélio pavimentoso estratificado ortoqueratinizado, de espessura delgada, com as características das camadas celulares equivalentes à mucosa jugal. A face ventral, todavia, exibiu-se com ausência de papilas, e a mucosa da face dorsal e ápice lingual de forma irregular, com papilas. Apresentou delgada camada de lâmina própria, com tecido conjuntivo frouxo na camada papilar e denso não modelado na camada reticular, quantitativo moderado de células, e expressiva concentração de fibras colagênicas, seguido de uma camada muscular bem desenvolvida. A comparação com a literatura mostrou forte semelhança entre os espécimes, com diferença mais marcante na presença de camada córnea no revestimento epitelial da mucosa jugal dos animais. Conclusão: A mucosa jugal e língua de ratos apresentaram semelhança à mucosa oral humana

Overview of bovine dendritic cells

This article is an overview of dendritic cells (DCs) in cattle. The understanding of the immune system and the role of DCs in many ways can contribute to their use in the prevention and treatment of many infectious and autoimmune diseases. DCs are bone marrow-derived cells that function as professional antigen presenting cells. They act as messengers between the innate and the adaptive immune systems. The morphology of DCs results in a very large surface to volume ratio. That is, the DCs have a very large surface area compared to the overall cell volume. Currently, most dendritic cells research occurs in the human and mice. There is a lack of studies in cattle describing DCs. DCs survey the body and collect information relevant to the immune system. They are then able to instruct and direct the adaptive arms to respond to challenges.O

Interventions for preventing oral mucositis for patients with cancer receiving treatment

Interventions for preventing oral mucositis for patients with cancer receiving treatmentTreatment for cancer (including bone marrow transplant) can cause oral mucositis (severe ulcers in the mouth). This painful condition can cause difficulties in eating, drinking and swallowing, and may also be associated with infections which may require the patient to stay longer in hospital. Different strategies are used to try and prevent this condition, and the review of trials found that some of these are effective. Two interventions, cryotherapy (ice chips) and keratinocyte growth factor (palifermin®) showed some benefit in preventing mucositis. Sucralfate is effective in reducing the severity of mucositis, and a further seven interventions, aloe vera, amifostine, intravenous glutamine, granulocyte‐colony stimulating factor (G‐CSF), honey, laser and antibiotic lozenges containing polymixin/tobramycin/amphotericin (PTA) showed weaker evidence of benefit. These were evaluated in patients with different types of cancer, undergoing different types of cancer treatment. Benefits may be restricted to the disease and treatment combinations evaluated

Altered maturation of peripheral blood dendritic cells in patients with breast cancer

Tumours have at least two mechanisms that can alter dendritic cell (DC) maturation and function. The first affects the ability of haematopoietic progenitors to differentiate into functional DCs; the second affects their differentiation from CD14+ monocytes, promoting an early but dysfunctional maturation. The aim of this study was to evaluate the in vivo relevance of these pathways in breast cancer patients. For this purpose, 53 patients with invasive breast cancer were compared to 68 healthy controls. To avoid isolation or culture procedures for enrichment of DCs, analyses were directly performed by flow cytometry on whole-blood samples. The expression of surface antigens and intracellular accumulation of regulatory cytokines upon LPS stimulation were evaluated. The number of DCs, and in particular of the myeloid subpopulation, was markedly reduced in cancer patients (P < 0.001). Patient DCs were characterized by a more mature phenotype compared with controls (P = 0.016), and had impaired production of IL-12 (P < 0.001), These alterations were reverted by surgical resection of the tumour. To investigate the possible role of some tumour-related immunoactive soluble factors, we measured the plasmatic levels of vascular endothelial growth factor, IL-10 and spermine. A significant inverse correlation between spermine concentration and the percentage of DCs expressing IL-12 was found. Evidence was also obtained that in vitro exposure of monocyte-derived DCs to spermine promoted their activation and maturation, and impaired their function. Taken together, our results suggest that both the above-described mechanisms could concomitantly act in breast cancer to affect DC differentiation, and that spermine could be a mediator of dysfunctional maturation of DCs

Numerical and functional defects of blood dendritic cells in early- and late-stage breast cancer

The generation of antitumour immunity depends on the nature of dendritic cell (DC)–tumour interactions. These have been studied mostly by using in vitro-derived DC which may not reflect the natural biology of DC in vivo. In breast cancer, only one report has compared blood DC at different stages and no longitudinal evaluation has been performed. Here we conducted three cross-sectional and one one-year longitudinal assessments of blood DC in patients with early (stage I/II, n=137) and advanced (stage IV, n=36) disease compared to healthy controls (n=66). Patients with advanced disease exhibit markedly reduced blood DC counts at diagnosis. Patients with early disease show minimally reduced counts at diagnosis but a prolonged period (1 year) of marked DC suppression after tumour resection. While differing in frequency, DC from both patients with early and advanced disease exhibit reduced expression of CD86 and HLA-DR and decreased immunostimulatory capacities. Finally, by comparing a range of clinically available maturation stimuli, we demonstrate that conditioning with soluble CD40L induces the highest level of maturation and improved T-cell priming. We conclude that although circulating DC are compromised by loco-regional and systemic breast cancer, they respond vigorously to ex vivo conditioning, thus enhancing their immunostimulatory capacity and potential for immunotherapy

Determinants of weight, psychological status, food contemplation and lifestyle changes in patients with obesity during the COVID-19 lockdown: a nationwide survey using multiple correspondence analysis

Introduction The corona virus disease 2019 (COVID-19) pandemic forced most of the Italian population into lockdown from 11 March to 18 May 2020. A nationwide survey of Italian Clinical Nutrition and Dietetic Services (Obesity Centers or OCs) was carried out to assess the impact of lockdown restrictions on the physical and mental wellbeing of patients with obesity (PWO) who had follow-up appointments postponed due to lockdown restrictions and to compare determinants of weight gain before and after the pandemic. Methods We designed a structured 77-item questionnaire covering employment status, diet, physical activity and psychological aspects, that was disseminated through follow-up calls and online between 2 May and 25 June 2020. Data were analyzed by multiple correspondence analysis (MCA) and multiple linear regression. Results A total of 1,232 PWO from 26 OCs completed the questionnaires (72% female, mean age 50.2 +/- 14.2 years; mean BMI 34.7 +/- 7.6 kg/m(2); 41% obesity class II to III). During the lockdown, 48.8% gained, 27.1% lost, while the remainder (24.1%) maintained their weight. The mean weight change was +2.3 +/- 4.8 kg (in weight gainers: +4.0 +/- 2.4 kg; +4.2% +/- 5.4%). Approximately 37% of participants experienced increased emotional difficulties, mostly fear and dissatisfaction. Sixty-one percent reduced their physical activity (PA) and 55% experienced a change in sleep quality/quantity. The lack of online contact (37.5%) with the OC during lockdown strongly correlated with weight gain (p &lt; 0.001). Using MCA, two main clusters were identified: those with unchanged or even improved lifestyles during lockdown (Cluster 1) and those with worse lifestyles during the same time (Cluster 2). The latter includes unemployed people experiencing depression, boredom, dissatisfaction and increased food contemplation and weight gain. Within Cluster 2, homemakers reported gaining weight and experiencing anger due to home confinement. Conclusions Among Italian PWO, work status, emotional dysregulation, and lack of online communication with OCs were determinants of weight gain during the lockdown period