Aims: To describe use of post-operative radiation for endometrial cancer in Ontario. To identify system-related and patient-related factors affecting access to this treatment. Materials and Methods: We performed a retrospective population-based cohort study of patients with surgically resected endometrial cancer in the Canadian province of Ontario between 1992-2003. Patients with evidence of incurable cancer at diagnosis or previous cancer diagnosis were excluded. We used multiple logistic regression to assess patient and system factors affecting radiation use. We controlled for disease-related and treatment-related factors: histology, surgical staging, type of hysterectomy and peritoneal biopsy. We applied a mixed model to account for clustering of data by operating hospital. Results: 9,411 women comprised the study cohort. The median age was 63 years. 26.2% received adjuvant radiation. The proportion of patients receiving radiation varied between cancer centre catchment areas from 18.0% to 34.3% (median 26.3%). In multivariate analysis, older patients were more likely to receive radiation up to the age of 80 (p<.0001). Patients who lived further from regional cancer centres were less likely to receive radiation (p=.0210). Patients who had their surgery during longer prevailing wait times at regional cancer centres were less likely to receive radiation (p=.0441). There was a 2.7-fold variation in the odds of radiation use between cancer centre catchments (p<.0001). Management at a comprehensive gynecologic oncology centre was associated with use of radiation for patients who had surgical staging of lymph nodes. Year of diagnosis and neighborhood income quintile did not significantly affect the use of radiation. Conclusions: There is wide variation in use of radiation for endometrial cancer in Ontario. There is evidence that system factors unrelated to patient’s needs affect use of adjuvant radiation for endometrial cancer in Ontario. Age is a key patient-related factor affecting radiation use. / Thesis (Master, Community Health & Epidemiology) -- Queen's University, 2009-08-07 22:02:37.308

The garlic extract ajoene is considered to have antimicrobial and antitumor effects against a variety of cell types, and it is suggested to have the potential to be used as an antifungal or antitumor drug clinically. The underlying mechanism of its inhibitory effects is still uncertain. In this project, the effects of ajoene on the growth of fungal and oomycete cells were studied on Candida albicans, Neurospora crassa and Achlya bisexualis. Endometrial cancer is the most common gynecologic cancer. A 3D spheroid model of endometrial cancer cells were for the first time used to investigate the antitumor effects of ajoene and selected antitumor agents. Ajoene was extracted from fresh garlic by chromatographic methods and the outcome of the extractions was verified with Mass spectrometry and NMR spectroscopy. Ajoene was then tested on the yeast form or germ tubes of C. albicans, and the cell division and germ tube formation was analyzed. N. crassa and A. bisexualis were treated with ajoene on plates or on glass slides to measure the hyphae radial extension or individual hyphal extension. 3D endometrial adenocarcinoma cell (Ishikawa) spheroids were treated with ajoene, paclitaxel, targeted drugs everolimus, sorafenib, gefitinib and canertinib alone or in combinations. The growth activity, metabolic activity, cell proliferation, apoptotic activity and the cytoskeletons were analyzed after the treatments. Cell division of C.albicans was inhibited by ajoene at 5µg/ml or higher concentrations. The length of C.albicans germ tubes was significantly shorter in ajoene treated groups than the untreated ones. Radial extension and individual hyphal extension of N. crassa and A. bisexualis were both inhibited by ajoene. Ajoene did not show any antitumor effects on the 3D cell model of Ishikawa cells. No synergistic effect was detected between ajoene and paclitaxel or ajoene and everolimus. The targeted drugs Canertinib and everolimus showed an inhibitory effect on growth activity of the spheroids, but no synergy with paclitaxel. In conclusion, ajoene was able to inhibit various forms of fungal and oomycete growth, but any antitumor activity of ajoene did not show on 3D culture of endometrial cancer cells.

Early diagnosis of endometrial cancer (EC) is uncertain and women undergo preventive hysterectomy in cases where a non-invasive treatment can be used instead. To contribute to solving this challenge we investigated if early changes in the nuclear 3D telomere architecture during carcinoma development can be detected prior to the first morphological evidence of precancerous lesions. We utilized Pten heterozygous mice that develop progressive carcinoma in the endometrial tissue similar to EC development in women. We used telomere fluorescence in situ hybridization (FISH), 3D molecular imaging and analysis techniques on interphase nuclei of endometrial glandular epithelial cells to identify alterations in the 3D-telomere profile. We found that telomere dysfunction in Pten heterozygous mice is present already in endometrial simple hyperplasia lesions prior to detectable loss of PTEN protein expression and that the 3D telomere architecture has a specific signature that indicates early telomere dysfunction predictive for endometrial malignant transformation.

Early diagnosis of endometrial cancer (EC) is uncertain and women undergo preventive hysterectomy in cases where a non-invasive treatment can be used instead. To contribute to solving this challenge we investigated if early changes in the nuclear 3D telomere architecture during carcinoma development can be detected prior to the first morphological evidence of precancerous lesions. We utilized Pten heterozygous mice that develop progressive carcinoma in the endometrial tissue similar to EC development in women. We used telomere fluorescence in situ hybridization (FISH), 3D molecular imaging and analysis techniques on interphase nuclei of endometrial glandular epithelial cells to identify alterations in the 3D-telomere profile. We found that telomere dysfunction in Pten heterozygous mice is present already in endometrial simple hyperplasia lesions prior to detectable loss of PTEN protein expression and that the 3D telomere architecture has a specific signature that indicates early telomere dysfunction predictive for endometrial malignant transformation.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

Diss. (sammanfattning) Umeå : Univ., 2004. / Härtill 6 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 5 uppsatser.

Thesis (Ph.D. in Reproductive Sciences) -- University of Colorado Denver, 2008. / Typescript. Includes bibliographical references (leaves 150-183). Free to UCD Anschutz Medical Campus. Online version available via ProQuest Digital Dissertations;

O objetivo geral deste trabalho foi caracterizar o perfil protéico da secreção uterina pura, através da eletroforese bi-dimensional, de éguas em diferentes fases do ciclo estral, com diferentes graus de alterações inflamatórias e degenerativas, e com diferente capacidade de resposta à cobertura. Para tanto se realizaram cinco experimentos. O primeiro objetivou caracterizar o perfil protéico da secreção endometrial de éguas e determinar se essas proteínas estão associadas ao ciclo estral. A hipótese a ser testada é que o perfil protéico das secreções endometriais varia entre o estro e o diestro. Conclui-se que a composição protéica da secreção uterina da égua em estro é diferente daquela de éguas em diestro, podendo as diferenças protéicas estarem associadas tanto ao processo de manutenção e desenvolvimento embrionário, como a uma necessidade de eventual resposta inflamatória. O segundo experimento objetivou comparar as taxas de prenhez e morte embrionária em éguas não lactantes cobertas no primeiro, ou em outros ciclos durante a temporada reprodutiva. Foi concluído que: (a) durante a temporada reprodutiva, a fertilidade de éguas solteiras é menor no 1° ciclo que nos ciclos subseqüentes; (b) que uma taxa de morte embrionária maior pode ser esperada em éguas falhadas cobertas no 1° ciclo do que nos demais e (c) que a menor fertilidade observada no 1° ciclo não está relacionada à incidência de endometrite persistente pós-cobertura. O terceiro experimento objetivou comparar o perfil protéico da secreção endometrial e a expressão de receptores para esteróides ovarianos no endométrio de éguas em diestro após a 1ª ovulação e após a 2ª ovulação da temporada. Conclui-se que o ambiente uterino é diferente no primeiro diestro da temporada reprodutiva, comparado aos ciclos subseqüentes. O quarto experimento objetivou comparar o perfil protéico da secreção endometrial de éguas resistentes e susceptíveis à endometrite persistente pós-cobertura, durante o estro.Ahipótese a ser testada é que o ambiente uterino da égua susceptível encontra-se alterado no estro prévio à cobertura. Conclui-se que a composição da secreção endometrial das éguas susceptíveis, antes da cobertura, é diferente daquela observada nas éguas resistentes, estando as proteinas observadas relacionadas ao processo inflamatório e à contratilidade uterina. O quinto experimento objetivou (a) comparar a composição protéica da secreção endometrial de éguas em estro com e sem fibrose e (b) avaliar o efeito da inflamação endometrial sobre o perfil protéico da secreção uterina de éguas em estro e em diestro. A hipótese é que a fibrose e a inflamação alteram a composição protéica do fluído uterino, podendo prejudicar o desempenho reprodutivo da égua. Conclui-se que nas éguas em estro a inflamação e a fibrose endometrial alteram apenas a quantidade de poucas proteínas no ambiente uterino. Durante o diestro, a presença da inflamação alterou o ambiente uterino, havendo maior quantidade de proteínas inflamatórias. Os resultados apresentados neste trabalho requerem a confirmação da identidade das proteínas, através de seqüeciamento ou imuno identificação. No entanto, várias informações puderam ser obtidas e muitas novas perguntas podem ser geradas, em especial a partir dos resultados com as éguas susceptíveis. / The main objective from this study was the characterization of the protein profile by SDS-PAGE, from mares during the estrous cycle and mares with uterine inflammation, fibrosis and susceptibility to persistent post-mating endometritis. Five experiments were performed. The first aimed to evaluate the protein profile of endometrial secretions of mares and to determine if any of these proteins was associated with estrous cycle. It was concluded that protein profile from estrous mares is different from the profile from diestrous mares. This difference can be explained by requirements to support and develop the embryo or to an eventually inflammatory response. The second experiment aimed to compare pregnancy and embryo loss rates in non-lactating mares bred either in the first, or in other estrus cycles during the breeding season. It was concluded that: (a) during the breeding the fertility of non-lactating mares is lower in the 1st than in other estrus cycles season; (b) that a higher embryo loss rate may be expected in barren mares bred in the 1st than in other estrus cycles of the breeding season and (c) that the lower fertility rate observed during the 1st estrus cycle is not related to the incidence of post-breeding endometritis. The third experiment aimed to compare the protein profile of endometrial secretions and the steroid hormone receptor expression of the uterus from diestrous mares in the first, or in other estrous cycles during the breeding season. It was concluded that the uterine environment in the first diestrus of the breeding season is different in relation of the other diestrus. The fourth experiment aimed to compare the protein profile of endometrial secretions from estrous mares resistant or susceptible to persistent post-mating endometritis (PPME). The tested hypothesis was that before the insemination in susceptible mares the uterine environment is disturbed. It was concluded that there is a difference in the uterine environmentbetween resistant and susceptible mares to PPME, probably resulting from the inflammatory response and affecting the uterine contractility. The fifth experiment aimed (a) to compare the protein profile of endometrial secretions from estrous mares with and without fibrosis and (b) to estimate the effect of inflammation in the protein profile of endometrial secretion from mares in estrous and in diestrous. The tested hypothesis was that fibrosis and inflammation disturb the uterine environment modifying the protein profile of the endometrial secretion impairing the reproductive performance. It was concluded that inflammation and fibrosis in estrous influenced the protein profile in a low number of spots. During diestrous, the inflammation affect uterine environment with an expressive number of inflammatory proteins. The protein profile observed in the experiments must be confirmed by sequencing or immunoidentification. However, much information could be obtained but many others must be investigated, principally by the susceptible mare.

A endometrite é uma importante causa de subfertilidade na égua. Infecções uterinas repetidas ou persistentes poderiam levar ao desenvolvimento de fibrose periglandular. Os objetivos deste trabalho foram avaliar se os processos degenerativos do endométrio são influenciados por infecções bacterianas experimentais sucessivas; realizar uma avaliação histopatológica das biópsias endometriais para documentar os efeitos de diferentes tratamentos sobre o endométrio após infecções experimentais. Foram utilizadas vinte éguas resistentes, com histórico reprodutivo desconhecido, e cinco éguas susceptíveis, com histórico de endometrites recorrentes e subfertilidade. As vinte e cinco éguas foram classificadas nos seguintes grupos, de acordo com o grau de endometrite e endometrose: GI (n=4), GIIA (n=10), GIIB (n=8) e GIII (n=3). Após o primeiro exame histopatológico (amostra pré-infecção), as éguas foram sincronizadas com prostaglandina. Na fase estral, as éguas foram infectadas com 1 x 109 Streptococcus equi subsp. zooepidemicus. Constatada a presença de sinais clínicos de endometrite, os grupos de éguas foram distribuídos entre cinco diferentes tratamentos: leucócitos frescos, leucócitos congelados, leucócitos lisados, Interleucina-8 (Il-8) e grupo controle.As éguas foram tratadas diariamente, por, no máximo, quatro dias, ou até que o exame bacteriológico não evidenciasse o crescimento de Streptococcus. No quinto dia, as éguas eram submetidas a novo exame histopatológico (amostra pós-infecção) e no sétimo dia, todas as éguas eram tratadas com penicilina, independentemente de terem eliminado a infecção ou não. Sete dias após, as éguas eram novamente submetidas a exame histopatológico e sincronizadaspara realizar uma nova infecção e novo tratamento. As biópsias foram avaliadas quanto à endometrose e endometrite. Não se observaram diferenças significativas entre os graus de endometrose observados antes e no quinto dia após as cinco infecções experimentais. Da mesma forma, não foram observadas variações no grau de endometrose entre as biópsias realizadas antes das infecções, bem como entre as biópsias coletadas no quinto dia após a infecção. Não houve influência dos tratamentos sobre os graus de endometrose durante as 5 infecções experimentais. Entretanto, verificou-se variação significativa entre as diferentes éguas no grau de endometrose das biópsias uterinas coletadas antes e após a infecção experimental. Observou-se, nas éguas resistentes, neutrofilia e eosinofilia significativas nas biópsias do 5º dia pós-infecção, quando comparadas com as biópsias pré-infecção. Nas éguas susceptíveis, somente foi detectada eosinofilia, não se observando aumento do número de neutrófilos. Não houve aumento do número de linfócitos e plasmócitos nas biópsias pós-infecção, quando comparadas às pré- infecção, nas éguas susceptíveis e resistentes. Os tratamentos não influenciaram, nas éguas resistentes e susceptíveis, a migração de neutrófilos no quinto dia, quando comparada com a observada antes da infecção. Conclui-se que infecções experimentais sucessivas durante 13 meses não influenciaram o grau médio de endometrose, apesar da variabilidade ocorrida entre as éguas. Esta variação provavelmente se deva a uma baixa representatividade de uma única amostra de biópsia na avaliação somente do grau de degeneração endometrial. Conclui-se, também, que éguas susceptíveis à endometrite, com presença de Streptococcus no útero, não apresentam neutrofilia cinco dias após a infecção. Provavelmente, o menor tempo de eliminação bacteriana observada nos tratamentos com leucócitos frescos e congelados deva-se a outros fatores que não o efeito quimioatraente leucocitário. / Endometritis is an important cause of subfertility in the mare. Repetitive or persistent uterine infections could lead to the development of periglandular fibrosis. The objectives of this study were: - to evaluate if degenerative endometrial diseases are influenced by repetitive experimental bacterial infections; - to determine the effect of different treatments on the endometrium after experimental infections by means of histopathological evaluation of endometrial biopsies. Twenty resistant mares, with unknown reproductive history, and five susceptible mares, with history of recurrent endometritis and subfertility, were used. Mares were classified, according to the degree of endometritis and endometrosis, within the following groups: GI (n=4), GIIA (n=10), GIIB (n=8) and GIII (n=3). Cycles were synchronized with prostaglandin after the first histopathological examination (pre-infection sample). During estrus, mares were infected with 1 x 109 Streptococcus equi subsp. zooepidemicus. Twenty four hours after the inoculation, clinical, bacteriological and cytological examinations were performed. When endometritis clinical signs were detected, the groups of mares were distributed into five different treatments: fresh leukocytes, frozen leucocytes, lysed leucocytes, Interleukin-8 (Il-8) and control group. Mares were treated on a daily basis for no more than four days, or until there was no Streptococcus growth in bacteriological examination. On the fifth day, mares were submitted to a new histopathological examination (post-infection sample) and, on the seventh day, all the mares were treated with penicillin, independently of having eliminated infection, or not. Seven days after, mares were submitted to a new histopathological examination and synchronized in order to proceed to a new infection and a new treatment. Biopsies were evaluated for endometritis and endometrosis. Therewas no significant difference regarding the degree of endometrosis before and on the fifth day after the five infections. Similarly, there was no difference regarding the degree of endometrosis among biopsies performed before infections and among biopsies performed on the fifth day after infections. Treatments did not influence the degree of endometrosis during the five experimental infections. Anyway, there was a significant variation between different mares, according to the degree of endometrosis, in biopsies collected before, as well as in biopsies collected after experimental infection. Resistant mares showed significant neutrophilia and eosinophilia in biopsies collected on the fifth day after infection, compared to pre-infection samples. Susceptible mares showed eosinophilia, but no growth in neutrophil number. There was no growth in lymphocyte and plasma cell number in post-infection biopsies, if compared to pre-infection biopsies, either in susceptible, or in resistant mares. In the same way, treatments did not influence neutrophil migration on the fifth day post-infection, if compared to pre-infection, either in susceptible, or in resistant mares. It was concluded that repetitive experimental infections do not influence the average degree of endometrosis, in spite of the variability between mares. This variability is probably due to the low representation of one single biopsy sample in the evaluation of the degree of endometrial degeneration. It was also concluded that susceptible mares showing Streptococcus in uterus do not present with neutrophil growth five days after infection. The shorter time required for bacterial elimination when fresh and frozen leukocytes were used is probably due to other factors than leukocyte chemoattractive effect.