Submitted by JÚLIO HEBER SILVA (julioheber@yahoo.com.br) on 2016-11-17T17:36:12Z No. of bitstreams: 2 Dissertação - Aparecida de Lourdes Carvalho - 2016.pdf: 1998821 bytes, checksum: 10b006baabfc797f656ed5a4e3660247 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Cláudia Bueno (claudiamoura18@gmail.com) on 2016-11-21T20:34:06Z (GMT) No. of bitstreams: 2 Dissertação - Aparecida de Lourdes Carvalho - 2016.pdf: 1998821 bytes, checksum: 10b006baabfc797f656ed5a4e3660247 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-11-21T20:34:06Z (GMT). No. of bitstreams: 2 Dissertação - Aparecida de Lourdes Carvalho - 2016.pdf: 1998821 bytes, checksum: 10b006baabfc797f656ed5a4e3660247 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-10-05 / CRISP-3 is a glycoprotein and a biomarker expressed at low levels by normal human prostate and strongly regulated in prostate cancer. The aim of this study was to investigate the presence of CRISP-3 biomarker in paraffin embedded prostate tissue specimens and to correlate the prognostic parameters pre and post treatment of prostate cancer patients. Cases diagnosed as prostate adenocarcinoma were selected from Centro de Prevenção de Jataí archives, from 2009 to 2013. Prostate specimens were analyzed histologically according to the Gleason score and the staging of prostate cancer was performed using the TNM system. The evaluation of clinical parameters was performed by searching the medical records of patients obtaining PSA data prior to surgery and during the follow-up. Immunohistochemical slides were evaluated by a pathologist and classified according to the intensity of staining / marking specific for CRISP-3. It was analyzed 25 tissue sections of prostate material and the expression of CRISP-3 protein was classified as strong in 14 (56%) patients, moderate in 4 (16%) and weak in 7 (28%) specimens. There was no correlation between the intensity of the imunohistochemical reaction and the levels of PSA pre and post treatment. The majority of the specimens, 24 (96%) were classified as usual acinar adenocarcinoma, 15 (60%) showed extension of the primary tumor pT2 and 13 (52%) had Gleason score equal to seven. In all analyzes there was no significant statistical differences among these parameters and the intensity of CRISP-3 staining. In this study, all prostate cancer analysed were positive for the presence of CRISP-3 suggesting the possibility of using this glycoprotein as an important biomarker for diagnosis of prostate cancer, especially at the time of diagnosis with the examination of needle biopsy, an important process in the active surveillance of this complication. / O CRISP-3 é uma glicoproteína e potencial biomarcador expressado em baixos níveis na próstata humana normal e fortemente regulado no câncer de próstata. O objetivo deste estudo foi investigar a presença do biomarcador CRISP-3 em espécimes de tecido prostático parafinizados e correlacionar com os atuais parâmetros prognósticos pré e pós tratamento de pacientes com câncer de próstata. Foram selecionados casos diagnosticados como adenocarcinoma prostático, dos arquivos do Centro de Prevenção de Jataí, no período de 2009 a 2013. Histologicamente analisou-se lâminas do material prostático de acordo com o score de Gleason e o estadiamento do adenocarcinoma prostático foi feito usando o sistema TNM. Fez-se avaliação dos parâmetros clínicos pela busca no prontuário médico dos pacientes obtendo-se os dados de PSA anterior à cirurgia e no segmento. Cortes histológicos corados por imunohistoquímica foram avaliados por médico patologista e classificados de acordo com a intensidade da coloração/marcação específica para CRISP-3. Foram analisados 25 cortes histológicos de material prostático quanto à imunoexpressão da proteína CRISP-3 sendo 14 (56%) de marcação forte, 4 (16%) moderada e 7 (28%) fraca. Não houve correlação entre a intensidade da reação imunohistoquímica e as dosagens bioquímicas de PSA pré e pós tratamento cirúrgico. A maioria dos espécimes, 24 (96%) foram classificados como adenocarcinoma acinar usual, 15 (60%) apresentaram extensão do tumor primário pT2 e 13 (52%) apresentaram score de Gleason igual a sete, em todas as análises não se observou diferença estatística significante entre os parâmetros analisados e a intensidade da marcação por CRISP-3. Neste estudo todas as análises foram positivas para a presença do CRISP-3 sugerindo a possibilidade de utilizar essa glicoproteína como importante biomarcador diagnóstico do câncer de próstata, principalmente na ocasião do diagnóstico com o exame de biópsia por agulha, processo importante na vigilância ativa dessa complicação.

The mare experiences a transient innate immune response to breeding, the resolution of which is crucial for optimal fertility. The majority of mares are able to modulate this inflammation in a timely fashion, but a subpopulation exists which fail to do so and are considered susceptible to persistent breeding-induced endometritis (PBIE). Seminal plasma has been shown to modulate aspects of this inflammation. Recently, two seminal plasma proteins have garnered interest for their immune modulating properties: cysteine-rich secretory protein-3 (CRISP-3) and lactoferrin. These proteins have been found to alter the binding between sperm and neutrophils based on sperm viability in vitro, but minimal work has evaluated their effect on endometrial mRNA expression of cytokines and inflammation in response to breeding. Experiments were performed to analyze the expression of equine CRISP-3. Found to be primarily synthesized in the ampulla of the vas deferens and to a lesser extent in the vesicular gland, CRISP-3 expression was only seen in the postpubertal stallion. Due to the effect of sperm viability on protein function in vitro, varying sperm populations were analyzed for their effect on gene expression in the uterus. It was determined that viable sperm suppressed the gene expression of the inflammatory modulating cytokine interleukin-6 (IL-6) in comparison to dead sperm. Next, the effect of CRISP-3 and lactoferrin on endometrial gene expression in the normal and susceptible mare was investigated. Neither protein had a significant effect on the mRNA expression of inflammatory cytokines in the normal mares at six hours post-breeding. In contrast, lactoferrin was found to significantly suppress the expression of the pro-inflammatory cytokine tumor necrosis factor (TNF)-α in susceptible mares. Due to this, lactoferrin was further analyzed as an immunomodulant for the treatment of PBIE. Susceptible mares were infused with varying doses of lactoferrin at six hours post-breeding. Although not in a dose-dependent fashion, lactoferrin was found to decrease both fluid retention and neutrophil migration, in addition to suppressing the expression of the pro-inflammatory cytokine interferon gamma (IFNγ) and increasing the gene expression of the anti-inflammatory cytokine interleukin-1 receptor antagonist (IL-1RN). In conclusion, CRISP-3 expression occurs in secretory aspects of the male reproductive tract, and appears to be up regulated after sexual maturation. Viability of spermatozoa affects the immune response to breeding and should be taken into consideration for experimental design and interpretation of data. The seminal plasma proteins CRISP-3 and lactoferrin have minimal effect on endometrial gene expression in normal mares, but lactoferrin suppresses the expression of TNF in susceptible mares. Finally, lactoferrin was found to function as a potent anti-inflammatory for the persistent inflammation seen in susceptible mares when administered post-breeding. This protein should be further investigated as a potential therapeutic for the treatment of persistent breeding-induced endometritis.