Targeting tumour cells usings chemical linkers with boronic acids and diazaborine scaffolds

Trabalho Final de Mestrado Integrado, Ciências Farmacêuticas, 2022, Universidade de Lisboa, Faculdade de Farmácia.O cancro é uma doença cuja prevalência tem vindo aumentar ao longo dos anos e que na maioria dos casos leva à morte do individuo que a possui. A sua elevada taxa de mortalidade, assim como os custos que encarrega, levaram a indústria farmacêutica e a comunidade académica a juntar-se de modo a encontrar novas soluções terapêuticas. O uso de fármacos citotóxicos com elevada potência é uma das soluções. Contudo, estes possuem reduzida seletividade, podendo ser perigosos para o organismo. Foram então desenvolvidas terapêuticas alternativas para solucionar este problema. Uma destas terapêuticas, é o uso de conjugados fármaco-anticorpo (ADCs). A estrutura dos ADCs está dividida em 4 componentes: um anticorpo, um fármaco citotóxico, um espaçador e um elemento de bioconjugação. As propriedades dos ADCs podem ser otimizadas alterando as estruturas dos seus componentes para que sejam mais eficazes e melhor tolerados em condições fisiológicas. Outra terapêutica também do nosso interesse é a criação de aglomerados proteicos com capacidade de encapsulamento do fármaco e de o transportar até ao interior das células alvo libertando o mesmo no seu interior. Têm-se vindo então a estudar diferentes estruturas que consigam ligar duas proteínas entre si, mas que sejam facilmente degradadas no citoplasma da célula tumoral. Para a realização deste projeto, tivemos como alicerce os ácidos borónicos (BAs), que são bastante versáteis devido à sua elevada reatividade. Estes permitem não só o desenvolvimento de ADCs, mas também de “linkers” inter-proteicos, devido à capacidade de se ligar a péptidos, assim como de reagir com hidrazinas para a formação de diazaborinas, moléculas estáveis em condições fisiológicas. Tivemos também em consideração a sua orbital p livre que permite reagir com espécies reativas de oxigénio (ROS) que se encontram em abundância em ambientes tumorais, e que levam à degradação destas moléculas. Durante este projeto focámo-nos então em planear e construir uma molécula bivalente que pudesse ser usada tanto como ADC assim como “linker” inter-proteico, que simultaneamente fosse sensível as condições fisiológicas especificas das células tumorais.Cancer is a disease whose prevalence has increased over the years and in most cases leads to the death of the individual who bears it. Its high mortality rate, as well as the costs involved in its maintenance have led the pharmaceutical industry and the academic community to come together in order to find new therapeutic solutions. The use of cytotoxic drugs with high potency is one of these solutions. However, the problem of these drugs is their reduced selectivity, which usually leads to the appearance of unwanted side effects. Different therapies were then developed to ameliorate this problem. One of these therapies is the use of antibody-drug conjugates (ADCs). The structure of ADCs is divided into 4 components: an antibody, a drug, a spacer and a bioconjugation technology. These bioconjugates attempt to solve the problem of selectivity by relying on the antibody’s high selectivity to deliver the payload safely to the intended site. ADCs properties can be optimized by changing the structures of their components so that they are as effective and tolerated as possible under physiological conditions. Another promising therapy is the creation of protein clusters capable of encapsulating the drug and transporting it to the interior of the target cells and releasing the drug in the interior. Different structures have already been reported which are able to bind two proteins together, and that are easily degraded in the cytoplasm of the tumour cell. On the idealization of this project, we considered boronic acids (BAs) as our foundation, as these molecules are quite versatile due to their high reactivity. They allow not only the development of ADCs, but also inter-protein linkers, due to their ability to bind with peptides, as well as react with hydrazines to form diazaborines, molecules that are stable under physiological conditions. We also took into account its free p orbital, which to reacts with reactive oxygen species (ROS) that are found in abundance in tumour environments, and lead to the degradation of BA´s. During this project, we focused on planning and building a bivalent molecule that could be used both as an ADC linker as well as an inter-protein linker, which was simultaneously sensitive to the specific physiological conditions of the tumour cells

Impactos do turismo na Região das Hortênsias-RS : a percepção do fenômeno pela comunidade local

Trabalho de Conclusão de Curso (graduação)—Universidade de Brasília, Instituto de Ciências Humanas, Departamento de Geografia, 2017.A paisagem, vista como uma categoria de análise/conceito pela Ciência Geográfica e apropriada pela Teoria do Turismo, vem a se transfigurar em produto turístico, sofrendo desta maneira diversos impactos, que devem ser compreendidos e mensurados para contemplar o devido diagnóstico e respectivo planejamento da atividade. A abordagem deste estudo refere-se aos impactos, sejam de ordem econômica, socioculturais e ambientais produzidos pela ação do turismo, bem como as respectivas degradações das paisagens nas áreas receptoras, tendo em específico como locus de pesquisa a Região das Hortênsias no Noroeste do Rio Grande do Sul, região típica de imigração alemã. A referida análise transita principalmente pela percepção da comunidade local, precípua interessada nos resultados da atividade em seu território, quanto aos possíveis benefícios ou mesmo ao prejuízo de seus próprios interesses.The landscape, seen as a category of analysis / concept by Geographic Science and appropriated by the Tourism Theory, is becoming a touristic product, suffering in this way several impacts, which must be understood and measured to contemplate the proper diagnosis and planning of the activity. The approach of this study refers to the economic, socio-cultural and environmental impacts produced by the tourism activity, as well as the respective degradations of the landscapes in the recipient areas, having in specific as locus of research the Region of Hydrangeas in the Northwest of Rio Grande do Sul, a region of German immigration. This analysis is mainly carried out by the local community, which is interested in the results of the activity in its territory, as to the possible benefits or even to the detriment of its own interests

Produção de espumante pelo Método Charmat com leveduras imobilizadas

Mestrado em Biotecnologia IndustrialA produção de vinho espumante tem sido alvo de intenso estudo e desenvolvimento nas últimas décadas. Partindo das mais recentes inovações no processamento deste vinho, neste trabalho estudou-se a cinética fermentativa de espumante em descontínuo pelo Método de Charmat, num fermentador de cuba fechada com recirculação, utilizando leveduras imobilizadas em alginato de cálcio (ProElif®). A primeira fase do trabalho, que precedeu os estudos cinéticos da fermentação, consistiu na caracterização física das esferas de ProElif® e do leito para determinação da velocidade mínima de fluidização, um dos parâmetros mais importantes quando se trabalha com fermentadores de leito fluidizado. Na segunda fase realizaram-se três estudos independentes para avaliação da influência de algumas variáveis na cinética fermentativa de vinho espumante. Avaliou-se o efeito do caudal de recirculação do vinho base e o efeito da dose de ProElif® aplicada. A cinética de consumo de açúcares foi bem descrita pelo modelo de Michaelis-Menten, tendo-se obtido um valor de KM médio de 25,2 g/L. O aumento da dose de esferas levou ao aumento da velocidade máxima da reação, como esperado. O estudo da influência do caudal de recirculação parece indicar que as limitações à transferência de massa externa são desprezáveis em relação internas neste sistema de células imobilizadas. Seguiu-se o estudo de estabilização tartárica do vinho base com aplicação da mais recente tecnologia de prevenção da precipitação dos sais de tartarato, a carboximetilcelulose (CMC). A CMC comprovou ser eficaz na estabilização tartárica do vinho para a dose máxima legal (10g/hL), mas mostrou ter influência na cinética do processo, verificando-se um aumento do tempo de fermentação. Por último, testou-se o reator batch sequencial (SBR), de forma a verificar a viabilidade de utilização das esferas de ProElif® em vários ciclos fermentativos consecutivos. Reutilizando-se as esferas durante oito ciclos consecutivos, verificou-se um aumento gradual da taxa máxima de consumo de substrato e uma diminuição do tempo total estimado de fermentação. O SBR poderá permitir o processamento de elevados volumes de vinho espumante, com muito menor tempo de operação e mão-de-obra associada.Sparkling wine production has been the subject of intense study and development for the last decades. Based on recent innovations, the aim of this work was to study the sparkling wine fermentation kinetics by Charmat Method, in a cuve close system with recirculation, using immobilized yeasts in a calcium alginate matrix (ProElif®). The first stage of this work consisted on physical characterization of ProElif® beads in order to determine the minimum fluidization velocity, one of the most important parameters in fluidized bed reactors. In the second phase three independent studies were done to assess the influence of some variables on the kinetics of fermentation of sparkling wine. Evaluation of the effect of recirculation flow rate of base wine and the effect of applied dose of ProElif® was made. The kinetics of sugar consumption was well described by the Michaelis-Menten model, yielding a value of average KM of 25,2g/L. As expected the dose of beads has led to increased reaction rates. The study of recirculation appears to indicate that the external mass transfer limitation are negligible in relation to the internal ones. The stabilization of wine base with application of latest technology to prevent the precipitation of tartrate salts, carboxymethylcellulose (CMC). The highest dose of CMC legal allowed (10g/hL) was effective in the stabilization of wine, but the process kinetics are slower, proving an increase in the fermentation time. Finally, in order to assess the feasibility of using ProElif® beads in several consecutive fermentations cycles, a sequential batch reactor (SBR) was assayed. Reusing the beads during eight consecutive cycles, promoted a gradual increase in the maximum rate of substrate consumption and a reduction of estimated fermentation time. Concluding, using immobilized yeast (ProElif®) in a cuve close system operated in SBR mode, allows processing of high volumes of sparkling wine, reducing operating time and manpower associated

Effects of isoflavones on the skin of postmenopausal women: a pilot study

OBJECTIVE: The aim of this study was to evaluate the effects of isoflavones on the skin of postmenopausal women. DESIGN: A prospective study was performed with 30 postmenopausal women before and immediately after the end of treatment with 100 mg/day of an isoflavones-rich, concentrated soy extract for six months. A skin punch was performed in the gluteal region for sample collection before and immediately after the treatment program. Morphometric determination of epidermal thickness, the papillary index (wrinkling), and the amount of dermal elastic and collagen fibers was assessed. In addition, the number of blood vessels in the sample was also evaluated. The paired Student's t-test was used for statistical analysis (P < 0.05). RESULTS: Isoflavone treatment resulted in a 9.46% increase in the thickness of the epidermis in 23 patients. In addition, the papillary index was reduced in 21 women. The papillary index was inversely proportional to skin wrinkling, i.e., there were a large number of papillae after treatment. The amount of collagen in the dermis was increased in 25 women (86.2%). In 22 women (75.8%) we observed that the number of elastic fibers increased. The number of dermal blood vessels was significantly increased in 21 women. CONCLUSION: Our data show that the use of a concentrated, isoflavone-rich soy extract during six consecutive months caused significant increases in epithelial thickness, the number of elastic and collagen fibers, as well as the blood vessels.Federal University of São Paulo Gynecology DepartmentFederal University of São Paulo Morphology DepartmentUniversidade de São Paulo Faculdade de Medicina Laboratório de Ginecologia Estrutural e MolecularUniversidade de São Paulo Faculdade de Medicina Departamento de Obstetrícia e GinecologiaUNIFESP, Gynecology DepartmentUNIFESP, Morphology DepartmentSciEL

Proliferation of the superficial epithelium of ovaries in senile female rats following oral administration of conjugated equine estrogens

OBJECTIVE: To evaluate the effect of different concentrations of estrogen on the ovarian superficial epithelium in senile female rats. Design: Fifty female rats at 15 months of age and with irregular estrous cycles were selected and randomly divided into five experimental groups containing equal numbers of animals in each: GPROP, control group receiving vehicle only; GE0.05mg, group receiving conjugated equine estrogens (CEE) at a dose of 50 µg/kg; GE0.5mg, group receiving CEE at 500 µg/kg; GE1mg, group receiving CEE at 1 mg/kg; and GE2mg, receiving CEE at 2 mg/kg. The length of treatment was 21 days. After this period, the animals were anesthetized and the ovaries were fixed in 10% formaldehyde and processed for routine histology. Histomorphology was analyzed by light microscopy, and histomorphometrics were evaluated using the Imagelab program. RESULTS: In the GPROP and GE0.05mg groups, the superficial epithelium of the ovary had a simple cuboidal shape, and as the estrogen dose increased, the epithelium thickened, with pseudo-stratified or stratified epithelium appearing in the GE2mg group. The animals in the group given the highest estrogen dose (GE2mg) showed the thickest ovarian epithelium and the largest perimeter and surface area of the surface ovarian epithelium (P < 0.01). However, the difference in epithelium thickness between the GE0.5mg and GE1mg groups was only slight. CONCLUSION: Our data suggest that CEE at a dose of 2 mg/kg may induce marked proliferation of rat ovarian epithelium.UNIFESP Morphology Department Histology and Cellular Biology DivisionUniversidade de São Paulo Faculdade de MedicinaUniversidade Federal de São Paulo (UNIFESP) Faculdade de Medicina Gynecology DepartmentUNIFESP, Morphology Department Histology and Cellular Biology DivisionUNIFESP, Faculdade de Medicina Gynecology DepartmentSciEL

Local anesthesia with epinephrine is safe and effective for oral surgery in patients with type 2 diabetes mellitus and coronary disease: a prospective randomized study

OBJECTIVE: To investigate the variations in blood glucose levels, hemodynamic effects and patient anxiety scores during tooth extraction in patients with type 2 diabetes mellitus T2DM and coronary disease under local anesthesia with 2% lidocaine with or without epinephrine. STUDY DESIGN: This is a prospective randomized study of 70 patients with T2DM with coronary disease who underwent oral surgery. The study was double blind with respect to the glycemia measurements. Blood glucose levels were continuously monitored for 24 hours using the MiniMed Continuous Glucose Monitoring System. Patients were randomized into two groups: 35 patients received 5.4 mL of 2% lidocaine, and 35 patients received 5.4 mL of 2% lidocaine with 1:100,000 epinephrine. Hemodynamic parameters (blood pressure and heart rate) and anxiety levels were also evaluated. RESULTS: There was no difference in blood glucose levels between the groups at each time point evaluated. Surprisingly, both groups demonstrated a significant decrease in blood glucose levels over time. The groups showed no significant differences in hemodynamic and anxiety status parameters. CONCLUSION: The administration of 5.4 mL of 2% lidocaine with epinephrine neither caused hyperglycemia nor had any significant impact on hemodynamic or anxiety parameters. However, lower blood glucose levels were observed. This is the first report using continuous blood glucose monitoring to show the benefits and lack of side effects of local anesthesia with epinephrine in patients with type 2 diabetes mellitus and coronary disease

Does Gestrinone Antagonize the Effects of Estrogen on Endometrial Implants Upon the Peritoneum of Rats?

OBJECTIVE: To evaluate the effects of estrogen treatment in combination with gestrinone on an experimental rat model of endometriosis. METHODS: Uterine transplants were attached to the peritoneum of female Wistar rats via a surgical autotransplantation technique. The implanted area was measured during the proestrus phase and after hormonal treatment. We performed morphometric analysis and examined the macroscopic and morphometric alterations of endometrial implants after hormonal treatment in ovariectomized rats. RESULTS: The high dose of estrogen caused macroscopic increases in the endometrial implant group compared with other groups, which were similar to increases in the proestrus phase. The low dose showed morphometric development of implants, such as an increase in number of endometrial glands, leukocyte infiltration and mitosis. Gestrinone antagonized both doses of estrogen. CONCLUSION: Our findings suggest that gestrinone antagonizes estrogen's effects on rat peritoneal endometrial implants

Review of magnetic resonance-guided focused ultrasound in the treatment of uterine fibroids

Uterine leiomyoma is the most frequently occurring solid pelvic tumor in women during the reproductive period. Magnetic resonance-guided high-intensity focused ultrasound is a promising technique for decreasing menorrhagia and dysmenorrhea in symptomatic women. The aim of this study is to review the role of Magnetic resonance-guided high-intensity focused ultrasound in the treatment of uterine fibroids in symptomatic patients. We performed a review of the MEDLINE and Cochrane databases up to April 2016. The analysis and data collection were performed using the following keywords: Leiomyoma, High-Intensity Focused Ultrasound Ablation, Ultrasonography, Magnetic Resonance Imaging, Menorrhagia. Two reviewers independently performed a quality assessment; when there was a disagreement, a third reviewer was consulted. Nineteen studies of Magnetic resonance-guided high-intensity focused ultrasound-treated fibroid patients were selected. The data indicated that tumor size was reduced and that symptoms were improved after treatment. There were few adverse effects, and they were not severe. Some studies have reported that in some cases, additional sessions of Magnetic resonance-guided high-intensity focused ultrasound or other interventions, such as myomectomy, uterine artery embolization or even hysterectomy, were necessary. This review suggests that Magnetic resonance-guided high-intensity focused ultrasound is a safe and effective technique. However, additional evidence from future studies will be required before the technique can be recommended as an alternative treatment for fibroids

ESTRATÉGIA COMO PROCESSO POLÍTICO: ENSAIO SOBRE A ALOCAÇÃO DE RECURSOS ORÇAMENTÁRIOS EM UNIVERSIDADES FEDERAIS NA PERSPECTIVA DAS RELAÇÕES DE PRODER

O objetivo deste estudo foi descrever e fomentar reflexões, acerca da formulação estratégica como um processo de tomada de decisão política no âmbito da distribuição interna de recursos orçamentários nas universidades públicas, gerando uma agenda para discussão sobre o tema. O método empregado neste ensaio teórico foi o descritivo-qualitativo com o intuito de propiciar a compreensão das particularidades e densidade do objeto de estudo. Foi utilizada a pesquisa bibliográfica para levantamento da teoria pertinente às relações de poder no processo de formulação estratégica nas organizações e a pesquisa documental para levantamento de informações quanto ao processo de distribuição orçamentária nas universidades federais brasileiras. Buscou-se sustentar a articulação da concepção do campo de poder proposto por Pierre Bourdieu, dos efeitos do exercício do poder no cotidiano das organizações, conceito preconizado por Michel Foucault e a amplitude dos dilemas organizacionais concebidos por Andrew M. Pettigrew. A partir das convergências dessas ideias foram estabelecidas interfaces entre política, poder e estratégia, inerentes a alocação interna de recursos orçamentários nas Universidades Federais. A proposta do ensaio foi promover contribuições para área e espera-se que as discussões levantadas sirvam de base para novos estudos que abordem essa temática que é relevante para a compreensão social das relações de poder nas estratégias organizacionais de instituições públicas e altamente complexas