Within the framework of this diploma, a genotypization of 162 beef cattle of the ČESTR commercial breed from was performed in order to analyze the potential influence of gene coding calpain I (CAPNI) on the resulting beef tenderness. CAPN1 present on the BTA29 chromosome, was selected in previous studies as a candidate gen for QTL. Polymorphism of CAPNI gene was studied using PCR/RFLP method and CAPN530 marker. Genotype identification resulted from this procedure. A 341 bp long fragment was present in homozygotes AA, two fragments of 195 and 146 bp were present in homozygotes GG, and three fragments of 341, 195 and 146 bp were detected in heterozygotes AG. The genotypization output was subsequently statistically evaluated. 11 homozygotes AA, 62 homozygotes GG and 89 heterozygotes AG were detected in the analyzed beef cattle population. More frequent G allele occurred in the set with the frequency of 0,657 and A allele with the frequency of 0,343. Warner-Bratzler shear test was employed to determine beef tenderness based on the shear force attribute. Samples of raw and grilled beef aging 1, 14 and 28 days after the slaughter were analyzed. Statistical methods were used to evaluate relationship between genotype and detected amount of shear force. In case of raw beef, only the samples aging 1 day after the slaughter showed significant difference in the shear force. The most positive results for the given parameter were achieved in case of AG genotype. Samples aging 14 and 28 days after the slaughter showed no difference in shear force and related beef tenderness, indicating no genotype influence. In case of grilled beef, no significant difference in sheer force indicating possible genotype influence was detected at any day of analysis. Genotype influence on grilled beef tenderness within the test animal population is minimal and statistically inconclusive.

O câncer de mama é a neoplasia responsável pelo maior número de mortes em mulheres no Brasil, portanto, é importante encontrar novos marcadores específicos e de diagnóstico precoce, utilizando procedimentos simples e rápidos. A via de sinalização Wnt regula importantes funções celulares como proliferação, sobrevida e adesão. Esta via está associada com os processos de iniciação e progressão em muitos tipos tumorais, como câncer de cólon familiar, melanoma e pulmão; sendo que mutações em β-Catenina (CTNNb1) explicam só 30% dos casos de sinalização aberrante encontrada no câncer de mama, indicando que existem outros componentes e/ou reguladores da via que possam estar envolvidos. O objetivo deste trabalho foi avaliar as variantes genéticas e epigenéticas nos genes do complexo de degradação de β-Catenina num grupo de pacientes com câncer de mama e num grupo controle; e determinar os perfis de transcrição dos componentes da via de sinalização Wnt e da molécula de expressão exclusiva do epitélio mamário, a Mamaglobina Humana (MGA), assim como associar estes resultados com as características clínicas, histológicas e patológicas do tumor. Para atingir este objetivo foram coletadas amostras de sangue periférico de 102 mulheres com câncer de mama e 102 mulheres sadias como grupo controle. A avaliação das variantes rs465899 do gene APC, rs2240308 e rs151279728 do gene AXIN2, rs5030625 do gene CDH1 e rs334558 do gene GSK3, foi realizada por meio de PCR-RFLPs e sequenciamento, a análise dos perfis de metilação dos promotores pela MS-PCR. A RT-qPCR foi usada para determinar os níveis de expressão dos componentes da via e a MGA. As variantes rs2240308 e rs151279728 do gene AXIN2 mostraram uma forte associação com o risco de desenvolver o câncer de mama. Um aumento significativo foi observado no nível de expressão de AXIN2 no grupo de mulheres com câncer de mama. Análises adicionais mostraram perfis de expressão diferencial dos genes APC, AXIN2, CTNNB1, GSK3 e CSNK1A1 associado ao status dos receptores hormonais e histogênese tumoral. MGA foi identificado exclusivamente em 38% dos pacientes com câncer de mama e foi associada com a progressão da doença. Este é o primeiro estudo que relaciona uma variante do gene AXIN2 com o câncer de mama na população brasileira. As variantes avaliadas do gene AXIN2 são marcadores promissores de susceptibilidade ao câncer de mama na população estudada, sendo importante, a avaliação desta variante genética na população e determinar o seu real efeito no processo de iniciação e/ou progressão do câncer de mama. / Background: Wnt/β-catenin signaling pathway is an important regulator of cellular functions such as proliferation, survival and cell adhesion. This pathway is associated with tumor initiation and progression; -catenin (CTNNB1) mutations explains only 30% of aberrant signaling found in breast cancer, indicating that other components and/or regulating of the Wnt/β-catenin pathway may be involved. Objective: The objective of the study was to evaluate the APC rs465899, AXIN2 rs2240308 and rs151279728, CDH1 rs5030625 and GSK3 rs334558 polymorphisms, APC, AXIN2, CDH1 and GSK3 promoter methylation status and expression profile of -Catenin destruction complex genes and MGA in peripheral blood of breast cancer patients. Methods: We collected peripheral blood samples from 102 breast cancer and 102 healthy subjects. The identification of the mutation was performed using PCR-RFLPs and DNA sequencing. MSP and HRM-MS was used to measure promoter methylation and RT-qPCR to determine expression profile. Results: We found significant association of AXIN2 rs2240308 polymorphism with breast cancer. Increased risk was observed even after stratification based on clinicpathological characteristics. AXIN2 rs151279728 polymorphism was found only in 9 breast cancer patients, but none in control group subject. APC and CDH1 polymorphisms were not associated with breast cancer. GSK3 polymorphism was weak associated with breast cancer and heterozygous status was associated with breast cancer protection after group stratification. APC and CDH1 promoter methylation in breast cancer patients was found. Significant increase was observed in AXIN2, CTNNB1 and GSK3 level expression in breast cancer patients. APC was down-regulated in breast cancer patients. Further analyses, showed APC, AXIN2, CTNNB1, GSK3 and CSKN1A1 gene expression associated to receptor status and histological type. MGA was found only in breast cancer patients and was associated with cancer progression. Conclusion: The present study reports, for the first time, that AXIN2 genetic defect and -catenin destruction complex expression disturbance may be found in breast cancer patients, providing additional support to the role of Wnt/-catenin pathway dysfunction in breast cancer tumorigenesis. However, the functional consequence of this genetic alteration remains to be determined. In another hand MGA was determined like a good biomarker for diagnosis and prognosis outcome.

O câncer de mama é a neoplasia responsável pelo maior número de mortes em mulheres no Brasil, portanto, é importante encontrar novos marcadores específicos e de diagnóstico precoce, utilizando procedimentos simples e rápidos. A via de sinalização Wnt regula importantes funções celulares como proliferação, sobrevida e adesão. Esta via está associada com os processos de iniciação e progressão em muitos tipos tumorais, como câncer de cólon familiar, melanoma e pulmão; sendo que mutações em β-Catenina (CTNNb1) explicam só 30% dos casos de sinalização aberrante encontrada no câncer de mama, indicando que existem outros componentes e/ou reguladores da via que possam estar envolvidos. O objetivo deste trabalho foi avaliar as variantes genéticas e epigenéticas nos genes do complexo de degradação de β-Catenina num grupo de pacientes com câncer de mama e num grupo controle; e determinar os perfis de transcrição dos componentes da via de sinalização Wnt e da molécula de expressão exclusiva do epitélio mamário, a Mamaglobina Humana (MGA), assim como associar estes resultados com as características clínicas, histológicas e patológicas do tumor. Para atingir este objetivo foram coletadas amostras de sangue periférico de 102 mulheres com câncer de mama e 102 mulheres sadias como grupo controle. A avaliação das variantes rs465899 do gene APC, rs2240308 e rs151279728 do gene AXIN2, rs5030625 do gene CDH1 e rs334558 do gene GSK3, foi realizada por meio de PCR-RFLPs e sequenciamento, a análise dos perfis de metilação dos promotores pela MS-PCR. A RT-qPCR foi usada para determinar os níveis de expressão dos componentes da via e a MGA. As variantes rs2240308 e rs151279728 do gene AXIN2 mostraram uma forte associação com o risco de desenvolver o câncer de mama. Um aumento significativo foi observado no nível de expressão de AXIN2 no grupo de mulheres com câncer de mama. Análises adicionais mostraram perfis de expressão diferencial dos genes APC, AXIN2, CTNNB1, GSK3 e CSNK1A1 associado ao status dos receptores hormonais e histogênese tumoral. MGA foi identificado exclusivamente em 38% dos pacientes com câncer de mama e foi associada com a progressão da doença. Este é o primeiro estudo que relaciona uma variante do gene AXIN2 com o câncer de mama na população brasileira. As variantes avaliadas do gene AXIN2 são marcadores promissores de susceptibilidade ao câncer de mama na população estudada, sendo importante, a avaliação desta variante genética na população e determinar o seu real efeito no processo de iniciação e/ou progressão do câncer de mama. / Background: Wnt/β-catenin signaling pathway is an important regulator of cellular functions such as proliferation, survival and cell adhesion. This pathway is associated with tumor initiation and progression; -catenin (CTNNB1) mutations explains only 30% of aberrant signaling found in breast cancer, indicating that other components and/or regulating of the Wnt/β-catenin pathway may be involved. Objective: The objective of the study was to evaluate the APC rs465899, AXIN2 rs2240308 and rs151279728, CDH1 rs5030625 and GSK3 rs334558 polymorphisms, APC, AXIN2, CDH1 and GSK3 promoter methylation status and expression profile of -Catenin destruction complex genes and MGA in peripheral blood of breast cancer patients. Methods: We collected peripheral blood samples from 102 breast cancer and 102 healthy subjects. The identification of the mutation was performed using PCR-RFLPs and DNA sequencing. MSP and HRM-MS was used to measure promoter methylation and RT-qPCR to determine expression profile. Results: We found significant association of AXIN2 rs2240308 polymorphism with breast cancer. Increased risk was observed even after stratification based on clinicpathological characteristics. AXIN2 rs151279728 polymorphism was found only in 9 breast cancer patients, but none in control group subject. APC and CDH1 polymorphisms were not associated with breast cancer. GSK3 polymorphism was weak associated with breast cancer and heterozygous status was associated with breast cancer protection after group stratification. APC and CDH1 promoter methylation in breast cancer patients was found. Significant increase was observed in AXIN2, CTNNB1 and GSK3 level expression in breast cancer patients. APC was down-regulated in breast cancer patients. Further analyses, showed APC, AXIN2, CTNNB1, GSK3 and CSKN1A1 gene expression associated to receptor status and histological type. MGA was found only in breast cancer patients and was associated with cancer progression. Conclusion: The present study reports, for the first time, that AXIN2 genetic defect and -catenin destruction complex expression disturbance may be found in breast cancer patients, providing additional support to the role of Wnt/-catenin pathway dysfunction in breast cancer tumorigenesis. However, the functional consequence of this genetic alteration remains to be determined. In another hand MGA was determined like a good biomarker for diagnosis and prognosis outcome.

The present study was to diagnose and classify the Human Papilloma Virus (HPV) in pregnant and non-pregnant adolescents, as study the risk factors related to the infection, identify the multiples oncogenics types presents in the study people and associate the differents types of HPV with cytology and colposcopy findings. Methodologic design: the type of study was prospective, descriptive and transversal coorte. Was study 111 sexually active adolescents, between 10 and 19 years old, patients of the Gynecology Service of the Professor Alberto Antunes University Hospital of the Federal University of Alagoas (HUPAA-UFAL). Was utilized the polymerase chain reaction / restriction fragment length polymorphism - sequencing (PCR/RFLPs) to identification of the virus. Results: the viral percentual in the inferior genital tract was 27% of the cases. The molecular genotyping revealed high risk viral genetic material - HPV 16, 33, 51, 58, 66, 1S39, CP8304 and LVX100, in 28,5% of the cases, low oncogenic risk - HPV 6, 11, 53, 61 e CP141, in 40% and, the other 31,4%, considered the indeterminate type, include high and low risk virus. This more incident type is found isolated or associated to other viral types and appears in 40% of the positive cases. The viral infection rate among the pregnant adolescents was 11,7%. The HPV genital infection was associated with a past of sexually transmitted diseases and the consumption of alcohol by the adolescent. The frequency of adolescents infected by HPV, associated to low grade intraepithelial lesion, was 5,0%. No associations were observed between the specific types of HPV and the cytology and colposcopy findings among the studied adolescents. / O objetivo do presente estudo foi para diagnosticar e classificar o papilomavírus humano (HPV) em adolescentes grávidas e não grávidas, bem como estudar os fatores de risco para a infecção, identificar os diversos tipos oncogênicos presentes na população estudada e associar os diferentes tipos de HPV com achados da citologia e colposcopia. Desenho metodológico: o tipo de estudo foi prospectivo, descritivo e de coorte transversal. Foram estudadas 111 adolescentes sexualmente ativas, entre 10 e 19 anos, atendidas no Serviço de Ginecologia do Hospital Universitário Prof. Alberto Antunes da Universidade Federal de Alagoas (HUPAA-UFAL). Utilizou-se a reação de polimerase em cadeia/polimorfismo de comprimento de fragmentos de restrição - sequenciamento (PCR/RFLPs), para a identificação do vírus. Resultados: o percentual do vírus no trato genital inferior foi de 27%. A genotipagem molecular revelou material genético viral de alto risco - HPV 16, 33, 51, 58, 66, 1S39, CP8304 e LVX100, em 28.5% dos casos, enquanto que os de baixo risco oncogênico - HPV 6b, 11, 53, 61 e CP141, em 40%. Os demais 31,4%, foram do tipo considerado indeterminado, que incluem vírus de alto e baixo risco. Esse tipo mais incidente encontra-se isolado ou associado a outros tipos virais e aparece em 40% do total dos casos positivos. A taxa de infecção viral entre as gestantes foi de 11,7%. A infecção genital pelo HPV teve associação com o passado de doenças sexualmente transmissíveis e com o consumo de álcool pela adolescente. A freqüência de adolescentes infectadas pelo HPV, associada à lesão intra-epitelial de baixo grau foi de 5,0%. Não foram observadas associações tipo-específicas do HPV com os achados da citologia e da colposcopia entre as adolescentes estudadas.