Le cancer du sein triple-négatif est un sous-type représentant environ 15 % des cancers du sein, qui est caractérisé par l'absence de récepteurs hormonaux aux œstrogènes et à la progestérone et l'absence d'expression du facteur de croissance HER-2. Le cancer du sein triple-négatif est également associé à un profil clinique défavorable avec un haut risque de rechute métastatique précoce. Actuellement, de nombreuses molécules sont testées comme cible thérapeutique dans ce type de cancer. La cathepsine D, une aspartyl protéase lysosome, est un marqueur de mauvais pronostic du cancer du sein associé au risque métastatique. Cette protéase est surexprimée par les cellules cancéreuses mammaires et hypersécrétée dans le microenvironnement tumoral. Notre équipe a apporté des contributions importantes pour élucider le rôle de la cath-D dans le cancer du sein. La cath-D stimule la prolifération des cellules cancéreuses, la croissance et l'angiogenèse tumorales ainsi que la formation des métastases. Au niveau moléculaire, le proenzyme sécrété par la cellule tumorale stimule la croissance invasive du fibroblaste mammaire via le récepteur LRP1 (low-density lipoprotein receptor-related protein-1). De plus, au pH extracellulaire acide des tumeurs, la pro-cathepsine D secrétée dégrade un des plus puissants inhibiteurs des cystéines cathepsines, la cystatine C, stimulant ainsi leurs activités protéolytiques.Mon travail a consisté à développer des anticorps anti-cath-D, présentant un effet thérapeutique dans le cancer du sein triple-négatif. Nous avons utilisé deux approches l'une par la technologie des hybridomes qui a permis d'isoler 50 anticorps, qui se sont avérés être des IgM. L'autre, par phage display, où 8 anticorps fragments scFv anti-cath-D ont été sélectionnés par ELISA. Ces 8 scFv ont été criblés pour leur capacité à bloquer l'activité biologique de la cathepsine D : inhibition de l'interaction avec LRP1 et inhibition du clivage d'un substrat fluorescent. Les anticorps E2 et F1 reformatés en IgG1 inhibent in vitro le phénomène de cicatrisation, la clonogénéicité, la croissance tridimensionnelle dans le matrigel et in vivo la croissance tumorale des cellules de cancer du sein triple-négatif MDA-MB-231. Cette étude est une première étape vers le développement de thérapie anti-cath-D dans le cancer du sein. / Cancer is the leading cause of death in the developed world. This project focuses t on the high incidence triple-negative breast cancers (ER- and PR-negative, HER2-non amplified) who present unsatisfactory treatments. Tumor progression is known to be due to cross-talk between cancer cells and their surrounding supporting tissue, the tumor stroma. The lysosomal aspartic protease cathepsin D is over-produced and abnormally hypersecreted by epithelial breast cancer cells. This protease is an independent marker of poor prognosis in breast cancer correlated with the incidence of clinical metastasis. Cathepsin D stimulates tumor progression by both proteolytic and non-proteolytic molecular mechanisms. Our team has shown that secreted pro-cathepsin D can alter the tumor microenvironment by interacting with partners such as the LRP1 (low-density lipoprotein receptor-related protein-1) receptor, and/or by cleaving endogenous substrates like cystatin C. My pHD project aimed to target extracellular procathepsin D released in breast cancer by developing specific therapeutic antibodies to block its catalytic and binding capacities. We have isolated 50 hybridomas, that secreted IgM, a pentameric antibody format with high avidity but low affinity, and 8 anti-cath-D scFv by phage display. The anti-human cath-D scFv were selected by functional screenings for their ability to inhibit both the proteolytic activity of Cath-D and its binding to the fibroblastic LRP1 receptor. Two scFv cloned under IgG1 format (F1 and E2) inhibited wound healing, colony formation and three-dimensional outgrowth in Matrigel of MDA-MB-231 triple-negative breast cancer cells, and their tumor growth in athymic nude mice. Our findings strongly suggest that antibody-based targeting of Cath-D within the breast tumor microenvironment may have therapeutic efficacy for breast cancer treatment.

This thesis presents the Knowledge Management Information Tool (KM-IT) Vendor Management Web Service and its reliability features. KM-IT is a web-based knowledge management information tool for the deactivation and decommissioning (D&D) user community. The Vendor Management (VM) module provides a directory service for searching D&D vendors. Clients have an increasing need to integrate and display the vendor information in their own applications. As such, this study proposes Web Services technology to provide VM access to clients. Moreover, when clients access information, particularly via mobile applications, they can encounter different failures that may occur on the network or the server. Such problems require fault tolerance in the VM Service. This study examines various reliability standards and selects the WS-ReliableMessaging standard as the best-suited approach to implement the reliability features for the VM Service. Implementation evaluation confirms that the KM-IT VM Service can effectively tolerate different types of failures.

L’aspartyl protéase cathepsine D, surexprimée et hyper-sécrétée par les cellules épithéliales cancéreuses mammaires est un facteur de mauvais pronostic des cancers du sein et stimule la prolifération des cellules cancéreuses et la formation des métastases. Elle affecte également le micro-environnement tumoral en induisant la croissance invasive des fibroblastes. Les travaux de l’équipe ont montré que le LDL-receptor related protein 1, LRP1, est le récepteur fibroblastique de la cathepsine D. LRP1 est fortement exprimé par les adipocytes. Les études cliniques indiquent que l’obésité est un facteur de risque dans de nombreux cancers, dont le cancer du sein chez la femme ménopausée.Dans cette thèse, nous avons étudié le rôle de la cathepsine D et du LRP1 dans les adipocytes, type cellulaire prédominant du micro-environnement tumoral mammaire. Nos résultats indiquent une surexpression de la cathepsine D et du LRP1 dans le tissu adipeux humain et murin obèse. De plus, l’expression de la cathepsine D est augmentée pendant la différenciation adipocytaire. Finalement, l’extinction de l’expression de la cathepsine D et du LRP1 inhibe l’adipogenèse indiquant leurs rôles clefs dans ce processus.L’ensemble de ces résultats suggère que la cathepsine D et son récepteur LRP1 pourraient être des cibles thérapeutiques potentielles dans le traitement de l’obésité. / The aspartyl protease cathepsin D, overexpressed and hyper-secreted by epithelial breast cancer cells is a poor prognosis factor in breast cancers and stimulates cancer cell growth and metastasis formation. It also affects the tumor microenvironment, inducing the fibroblasts invasive outgrowth. Our works have shown that the LDL receptor-related protein1, LRP1, is the fibroblastic receptor for cathepsin D. LRP1 is highly expressed in adipocytes. Clinical studies indicate that obesity is a risk factor in many cancers, including breast cancer in postmenopausal women.During this thesis, we studied the role of cathepsin D and LRP1 in adipocytes, which are the prominent cell type in the tumor microenvironment of breast cancers. Our results indicate that cathepsin D and LRP1 are overexpressed in human and mouse obese adipose tissue. Furthermore, the expression of cathepsin D is increased during adipocyte differentiation. Finally, the inhibition of the cathepsin D and LRP1 expression inhibits adipogenesis indicating their key role in this process.All these results suggest that cathepsin D and its receptor LRP1 could be potential therapeutic targets in the treatment of obesity.

Orientadores: Denise Engelbrecht Zantut Wittmann, Sarah Monte Alegre / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-28T02:28:01Z (GMT). No. of bitstreams: 1 Botelho_IlkaMaraBorges_M.pdf: 1340793 bytes, checksum: 6ef5a41b67a7eddca52fc55dea2f96d4 (MD5) Previous issue date: 2014 / Resumo: Introdução: Vitamina D tem sido apontada como importante regulador da resposta imune. Estudos tem demonstrado haver relação entre insuficiência de vitamina D e presença de doenças autoimunes como Tireoidite de Hashimoto (TH). É possível que o processo autoimune na TH seja inibido em diferentes estágios pela vitamina D em sua forma ativa. Nossos objetivos foram estudar a prevalência de insuficiência de vitamina D e a relação de suas concentrações séricas com marcadores de função e autoimunidade tireoideana. Material e Métodos: Amostras de sangue foram coletadas de 54 pacientes com TH e 54 indivíduos saudáveis sem diagnóstico de TH com idade entre 18 e 75 anos. Foram realizadas dosagens séricas de vitamina D (25OHD), TSH, T4 livre, cálcio, fósforo, paratormônio (PTH), anticorpos anti-tireoperoxidase (AcTPO), anti-tireoglobulina (AcTG) e anti-receptor de TSH (TRAb). Volume tireoideano foi estimado por ultrassonografia. Foram coletados dados demográficos, de peso, altura, índice de massa corporal (IMC) e tempo de diagnóstico. Pacientes e indivíduos do grupo de controle foram pareados por idade e sexo. O nível de significância estatística adotado foi 5%. Resultados: Prevalência de insuficiência de vitamina D foi encontrada em 68.5% dos pacientes e em 38.9% dos indivíduos do grupo de controle (p =0,002). Houve uma correlação positiva entre níveis de AcTPO e maior volumetireoideano nos pacientes (r = 0,319; p= 0.019). Não houve correlação entre concentração de vitamina D, TSH, T4livre,TRAb, AcTGe volume tireoideano. Conclusões: Demonstramosmaior prevalência deinsuficiência de vitamina Dem pacientescom tireoidite de Hashimotoem relaçãoa indivíduos de um grupo controlesaudável, não havendo correlaçãocom o estado hormnal tireoideanooumarcadores séricos deautoimunidadeda tireóide.Por sua vez, maior volume da tireóidese associou a maior grau de infiltração inflamatóriaautoimune,refletido pelacorrelaçãocom maiores concentrações AcTPO / Abstract: Introduction: Vitamin D has been pointed out as an important immune response regulator. Studies have shown a relationship between vitamin D insufficiency and the presence of autoimmune diseases such as Hashimoto's Thyroiditis (HT). It's possible that the autoimmune process in HT is inhibited in its different stages by vitamin D on its active form .Our aims were to study the prevalence of vitamin D insufficiency and relationship of the serum concentrations with thyroid function and autoimmunity markers. Material and Methods: Blood samples were collected from 54 patients with HT and 54 healthy individuals without a diagnosis of HT, aged 18 to 75 years. We conducted serum 25OH vitamin D, TSH, free T4, calcium, phosphorus, PTH, TPOAb, TgAb and TRAb. Thyroid volume was estimated by ultrasound. Data on demographic, weight, height, body mass index and time since diagnosis were collected. Patients and control subjects were matched by sexand age. The significance level for statistical analysis was 5%. Results: Prevalence of vitamin D insufficiency was found in 68.5% of patients and in 38.9% of subjects in the control group (p= 0.002). There was a positive correlation between TPOAb and volume in patients (p= 0.019). There was no correlation between vitamin D concentration and thyroid volume, TRAb, TgAb, TSH or free T4. Conclusions: We demonstrated a higher prevalence of vitamin D insufficiency in patients with Hashimoto's thyroiditis compared to individuals of a healthy control group, no correlation with thyroid state hormonal or serum markers of thyroid autoimmunity. In turn, greater thyroid volume was associated with a higher degree of autoimmune inflammatory infiltration, reflected by the correlation with higher concentrations AcTPO / Mestrado / Clinica Medica / Mestra em Ciências

Multiple sclerosis (MS) is a progressive, autoimmune neurodegenerative disorder affecting nearly 350,000 people in the United States and resulting in significant disability. As an immunomodulator, vitamin D may play a role in the development of MS. Previous studies have observed an inverse association of 25-hydroxyvitamin D (25(OH)D) levels and MS risk in younger populations; however, whether this relationship persists in older adults remains unclear. We prospectively investigated the association between predicted 25(OH)D level and incident MS in the Nurses’ Health Study (NHS) (n=121,701) and NHS II (n=116,430). 25(OH)D levels were predicted using validated regression models that include important determinants of vitamin D status, including race, UV-B flux (based on state of residence), physical activity, body mass index, dietary vitamin D intake, alcohol consumption and post-menopausal hormone use. Data on these factors were self-reported on NHS and NHS II questionnaires starting in 1986 and 1991, respectively, and updated every 2-4 years. MS diagnoses were ascertained by self-report and confirmed by medical records. Cox proportional hazards models adjusted for age, ethnicity, latitude of residence at age 15, and BMI at age 18 were used to estimate hazard ratios (HR)s and 95% confidence intervals (CI)s in each cohort. During up to 18 years of follow-up, we documented 179 definite/probable cases of MS with first symptoms after baseline. Multivariable HRs comparing highest and lowest quintiles of predicted 25(OH)D were 1.09 (95% CI: 0.40-2.96) in the NHS and 0.52 (95% CI: 0.28-0.95) in the NHS II. Higher predicted plasma 25(OH)D may be modestly associated with lower risk of MS, primarily in younger women.

Objectives: In recent years, vitamin D deficiency has been recognized increasingly often in patients, and different supplement regimens have been prescribed to treat it. There has been no consensus on treatment regimens. This study was conducted to determine the management of vitamin D deficiency/insufficiency in outpatient adults in northeast Tennessee. Methods: A retrospective record review was conducted in an internal medicine teaching clinic for patients seen from July 2007YJuly 2008 in Johnson City, Tennessee. A total of 626 nonelectronic charts listed with vitamin D measurements were used in the analysis. Data regarding the level of vitamin D, whether treatment was prescribed, dose and duration of treatment prescribed, and repeat levels of vitamin D, if any were ordered, were collected. Vitamin D deficiency was defined as levels G20 ng/mL; vitamin D insufficiency was defined as levels ranging from 20 to 29.9 ng/mL. Results: Of the 626 patients, 325 (52%) were vitamin D deficient or insufficient. Of these 325 patients, 184 were given a low-dose supplement and 54 received a high-dose supplement. Eighty-seven were either not prescribed any replacement or the dose was unknown (not documented in the chart). The mean change in serum vitamin D levels was significantly different for the high dose compared with the low dose prescribed. There was no significant sex difference in response to the dose given. On average, those who were vitamin D deficient experienced a greater change than those who were insufficient and a greater change, on average, was observed in those who received a higher dose. Conclusions: Vitamin D deficiency and insufficiency are highly prevalent. Clinicians tended to prescribe a high dose of treatment for lower levels of serum vitamin D. The response is higher in high-dose treatment. Documentation regarding whether vitamin D supplements were given or the dose of supplements was given and followed up with repeat levels of vitamin D after treatment was poor.

BACKGROUND:: Vitamin D deficiency is increasingly being recognized as a highly prevalent and undertreated problem. This study was conducted to determine the prevalence of vitamin D deficiency/insufficiency in hospitalized adults in northeast Tennessee. METHODS:: A prospective cohort study was conducted on 99 inpatients admitted to an internal medicine teaching service from July through October 2006 at a single private hospital in Johnson City, Tennessee. A single measurement of 25-hydroxyvitamin D was performed on all patients. RESULTS:: Of the 99 patients, 53% were vitamin D deficient or insufficient (30% deficient with a level of <20 ng/mL and 23% insufficient with a level between 20ĝ€"29.9 ng/mL). The highest frequency of deficiency was in females <50 years. CONCLUSION:: Vitamin D deficiency is highly prevalent in all age groups and in both females and males in this population. Clinicians should consider measuring the vitamin D level of all inpatients on a routine basis.

No description available.

Although maternal vitamin D status has been linked to asthma in offspring, the relationship between vitamin D status and asthma in adults still remains unclear. The current study assessed the relationship between measures of vitamin D status and self-reported asthma/wheeze in 186 healthy women aged 18-30 years. Although the risk of asthma/wheeze symptoms was three-times higher among women with low dietary vitamin D intake (<200 IU>/day) than in those with higher vitamin D intake, suboptimal serum levels of 25(OH)D ( <70 nmol>/L) were associated with a 48% lower risk of asthma/wheeze than “optimal” serum levels. These contradictory effects underscore the poor correlation between dietary vitamin D intake and serum vitamin levels and suggest that other components in vitamin D-rich foods may be protective. Alternatively, women with higher serum vitamin D levels may have spent more time outdoors, increasing their exposure to asthma triggers. This study also identified predictors of serum 25 (OH) D in this sample. In addition to total dietary vitamin D (r= 0.2; p=0.03), intake of cold cereal (p=0.02) also significantly predicted serum 25(OH)D levels. Among non-dietary factors, month of blood draw (p=0.05) and oral contraceptive use (p<0.0001) were positive predictors of serum 25(OH) D; sunscreen use (p=0.04) was a negative predictor. After adjusting for covariates, oral contraceptive use was associated with 25(OH)D levels that were on average 24 nmol/L greater than those observed in women who did not use oral contraceptives. Additional prospective studies are needed to further evaluate the relationship between vitamin D status and asthma.

No description available.