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Orientador: Sarah Arana / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-15T00:25:10Z (GMT). No. of bitstreams: 1 Tagliaferro_AlineFernanda_M.pdf: 13833602 bytes, checksum: 8ff9e6fc1f703f847914ff9a4284c900 (MD5) Previous issue date: 2009 / Resumo: Dado o registro de endosulfan nas bacias do Pantanal Mato-Grossense, o risco para os organismos aquáticos que esse agrotóxico promove e a falta de informações sobre sua ação na ictiofauna dessa região, esta pesquisa objetivou verificar a sensibilidade do pacu, Piaractus mesopotamicus, ao endosulfan. Para a realização da intoxicação aguda (96h) em sistema estático, alevinos foram expostos às seguintes concentrações de endosulfan: 0; 0,71; 1,43; 2,14; 2,86; 3,57; 4,29 e 5µgL-1. Após 96h, os exemplares sobreviventes foram necropsiados e amostras de brânquia, fígado e rim foram colhidas para análise histopatológica, qualitativa e semi-quantitativa. A Concentração Letal Média (CL50) obtida foi de 5,66µgL-1 (24h) e 4,33µgL-1 (96h). Alterações branquiais e hepáticas foram perceptíveis à microscopia de luz (ML) e à microscopia eletrônica de transmissão (MET), já no rim somente alterações à MET foram detectadas. Nas brânquias, lamelas secundárias com hipertrofia do epitélio, telangectasia e destacamento epitelial foram as principais alterações. Já no fígado notaram-se degeneração hidrópica, inclusão nuclear, inclusão hialina e vacuolização citoplasmática, como as alterações mais frequentes. Entre as alterações à MET, de modo geral, as mais frequentes foram: alterações mitocondriais, presença de figura de mielina, alteração e/ou perda de microvilos e aumento do espaço intercelular. Embora, alguns exemplares apresentaram alterações celulares frequentes e severas, sugerindo sério dano induzido pelo endosulfan, a análise semi-quantitativa indicou grande variabilidade interindividual. Este último dado sugere fortemente, que a avaliação de biomarcadores histológicos de contaminação aquática deve ser acompanhada de avaliação semi-quantitativa das alterações induzidas associada à análise estatística, para uma fiel indicação do dano provocado à população de organismos-teste. Finalmente, o pacu mostrou-se um bom bioindicador de contaminação aquática por endosulfan o qual se mostrou extremamente tóxico para essa espécie. Assim, espera-se que esta pesquisa venha contribuir efetivamente para o estabelecimento de normas mais rigorosas, ou até mesmo a proibição, da utilização do endosulfan em território brasileiro à semelhança de outros países. / Abstract: The records of endosulfan in river basins of the Patanal wetlands in the state of Mato Grosso (Brazil), the risk to aquatic organisms and lack of information on the action of this pesticide on fish fauna in this region stimulated the present study, which aim was to determine the sensitivity of the pacu (Piaractus mesopotamicus) to endosulfan. For the acute intoxication test (96h) in a static system, fingerlings were exposed to the following concentrations of endosulfan: 0; 0.71; 1.43; 2.14; 2.86; 3.57; 4.29 and 5µgL-1. After 96h, the specimens were necropsied and samples were taken from the gill, liver and kidney for qualitative and semi-quantitative histopathological analysis. The Mean Lethal Concentration (LC50) was 5.66µgL-1 (24h) and 4.33µgL-1 (96h). Gill and liver alterations were visible through light microscopy and transmission electron microscopy (TEM); alterations in the kidney were only detected through TEM. The main alterations in the gills were secondary lamellae with epithelial hypertrophy, telangiectasis and epithelial lifting. The most frequent alterations in the liver were hydropic degeneration, hyaline inclusion and cytoplasmic vacuolization. The most frequent TEM findings were mitochondrial alterations, the presence of myelin figure, alterations in and/or loss of microvilli and increase in intercellular space. Although some specimens frequently exhibited severe cell alterations suggesting serious damage induced by endosulfan, the semi-quantitative analysis revealed considerable interindividual variability. This strongly suggests that the evaluation of histological biomarkers of aquatic contamination should be accompanied by a semi-quantitative assessment of alterations associated with statistical analysis in order to obtain a faithful indication of the damage caused to the population of test organisms. The pacu proved to be a good bioindicator of aquatic contamination by endosulfan, which proved to be extremely toxic to this species. It is hoped that the present study can effectively to contribute toward the establishment of stricter norms or even the banning of the use of endosulfan in Brazil, as has occurred in other countries. / Mestrado / Histologia / Mestre em Biologia Celular e Estrutural

Background: New prognostic grade groupings were recently proposed for prostate cancer. They are based on Gleason grading of either biopsy or prostatectomy specimen. Former Gleason 6 corresponds to group 1, Gleason 7=3+4 corresponds to group 2, Gleason 7=4+3 corresponds to group 3, Gleason 8 corresponds to group 4, and Gleason 9-10 correspond to group 5. Objective: To assess the prognostic value of Gleason grade groups in men with prostate cancer from a nationwide population‑based cohort. Design, Setting and Participants: From the National Prostate Cancer Register of Sweden, we identified 5,880 men diagnosed with prostate cancer from 2005 to 2007, including 4,325 who had radical prostatectomy and 1,555 treated by radiotherapy.  Outcome Measurements and Statistical Analysis: Kaplan-Meier survival analysis was used to calculate the cumulative 4-year biochemical recurrence-free survival. Cox proportional hazards regression models were used to examine the relationship between prognostic Gleason grade groups and biochemical recurrence after radical prostatectomy and radiotherapy. The 4-year biochemical progression-free survival was compared for groups based on biopsy and prostatectomy Gleason grade groups. Results and Limitations: Among men undergoing surgery, the 4‑year biochemical progression-free survival was 89%, 82%, 74%, 77%, and 49% for prognostic Gleason grade groups 1-5 on biopsy. The corresponding 4-year biochemical progression-free survival based on prostatectomy prognostic Gleason grade groups was 92%, 85%, 73%, 63%, and 51% for prognostic Gleason grade groups 1-5. For men undergoing radiotherapy, biopsy prognostic Gleason grade groups 1-5 had 4-year biochemical progression-free survival of 95%, 91%, 85%, 78%, and 70%. After adjusting for preoperative serum prostate specific antigen and clinical stage, biopsy prognostic Gleason grade groups were significant independent predictors of biochemical progression after radical prostatectomy and radiotherapy. There was no central review of pathology. Conclusions: These results confirm the prognostic value of the newly proposed prognostic Gleason grade groups in men undergoing radical prostatectomy and radiotherapy in a population-based setting.

Biological and molecular characteristics of the pathogenic mollicute, Spiroplasma penaei, isolated from the hemolymph of infected Pacific white shrimp, Penaeus vannamei, were investigated. The doubling times of a S. penaei were 6.13 h (2% NaCl) and 3.43 h (no salt) under aerobic conditions, and 6.63 h (2% NaCl) and 3.22 h (no salt) under anaerobic conditions. Small diffuse white colonies with granular centers, surrounded by small satellite colonies that appeared embedded in the agar matrix, were detected on solid M1D medium (2% Noble agar) under aerobic conditions at 28°C. The genome size of the S. penaei was 1778 Kb, as determined by pulsed-field gel electrophoresis using undigested DNA. Reduction of virulence of S. penaei was not detected in serial passage 24 and 76 isolates but passage 131 isolate was attenuated as indicated by the number of surviving shrimp and histological findings of challenged P. vannamei. Toxicity was not detected in supernatant fractions of M1D medium cultures of S. penaei isolates. The most predominant host responses to the S. penaei reference isolate and to serial passage isolates were hemocytic nodules and hemocytic infiltration observed in hematoxylin and eosin-stained histological sections. Transmission electron microscopy of the lymphoid organ of experimentally infected P. vannamei depicted S. penaei without cell wall and free in the cytoplasm of lymphoid organ cells. The lesions observed in histological sections were verified by in situ hybridization using a digoxigenin (DIG)-labeled probe specific to the spiralin gene of Spiroplasma spp. Evolutionary relationship trees, based on five partial DNA sequences of 16S rDNA, 23S rDNA, 5S rDNA, gyrB, rpoB genes and two complete DNA sequences of 16S-23S and 23S-5S ISR, were constructed using the distance-based Neighboring-Joining method with Kimura-2-parameter substitution model. The NJ trees based on all DNA sequences investigated in this study positioned S. penaei in the Citri-Poulsonii clade and corroborates the observations by other investigators using the 16S gene. Pairwise genetic distance calculation between sequences of spiroplasmas showed S. penaei to be closely related to S. insolitum and distantly related to Spiroplasma sp. SHRIMP from China.

Intraductal papillary mucinous neoplasms (IPMN) are pancreatic lesions with uncertain biologic behavior. This study sought objective, accurate prediction tools, through the use of quantitative histopathological signatures of nuclear images, for classifying lesions as chronic pancreatitis (CP), IPMN, or pancreatic carcinoma (PC). Forty-four pancreatic resection patients were retrospectively identified for this study (12 CP; 16 IPMN; 16 PC). Regularized multinomial regression quantitatively classified each specimen as CP, IPMN, or PC in an automated, blinded fashion. Classification certainty was determined by subtracting the smallest classification probability from the largest probability (of the three groups). The certainty function varied from 1.0 (perfectly classified) to 0.0 (random). From each lesion, 180 +/- 22 nuclei were imaged. Overall classification accuracy was 89.6% with six unique nuclear features. No CP cases were misclassified, 1/16 IPMN cases were misclassified, and 4/16 PC cases were misclassified. Certainty function was 0.75 +/- 0.16 for correctly classified lesions and 0.47 +/- 0.10 for incorrectly classified lesions (P = 0.0005). Uncertainty was identified in four of the five misclassified lesions. Quantitative histopathology provides a robust, novel method to distinguish among CP, IPMN, and PC with a quantitative measure of uncertainty. This may be useful when there is uncertainty in diagnosis.

As neoplasias são doenças comumente vistas em algumas espécies de aves, porém, no Brasil, não existem trabalhos publicados sobre casuística e classificação destes processos nas mesmas. Com o aumento da criação de aves como animais de companhia, torna-se importante o aprimoramento do conhecimento na área. Este estudo, realizado de forma retrospectiva englobando os anos de 2000 a 2004 e prospectiva nos anos de 2005 e 2006 no Ambulatório de Aves do Hospital Veterinário, e no Departamento de Patologia da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, teve como objetivos estudar a freqüência e a prevalência com que ocorrem os processos neoplásicos em aves, e realizar avaliação destes através da histopatologia e imunoistoquímica, comparando com os achados de literatura. Os resultados concordaram em parte com os achados em literatura sendo o periquito-australiano a ave com a maior prevalência e o lipoma sendo a neoplasia de maior ocorrência. Indica-se o uso da imunoistoquímica, porém, deve-se aprimorar estudos histopatológicos e investir em diagnóstico para o melhor conhecimento da biologia tumoral, desenvolvendo-se mais pesquisas na área. / Neoplastic diseases are generally seen in some avian species, nevertheless, in Brazil, there are no reports regarding cause and classification of these diseases in birds. As keeping birds as pets is increasing, it is important to upgrade knowledge in this area. This report, made retrospectively in 2000 to 2004 and prospectively in 2005 and 2006 in the Avian Clinic of Veterinary Teaching Hospital, Department of Pathology, School Veterinary Medicine of University of São Paulo, has the objectives of studying the frequency and prevalence of neoplastic diseases in birds, and evaluating them by histopathology and immunohistochemistry, comparing results with literature. The results agree in part with those found in literature, the budgerigar is the most prevalent specie and lipoma as the most common neoplasm. The use of immunohistochemistry is indicated, but more studies regarding histopathology and investment are necessary in diagnosis and research in tumoral biology, developing more researches in this area.

Orientador: Isabel Cristina Cherici Camargo / Co-Orientador: Luiz Gustavo de Almeida Chuffa / Banca: Telma Gonçalves Carneiro Spera de Andrade / Banca: Francisco Eduardo Martinez / Resumo: Embora sejam extensas as opções de aplicação terapêutica dos esteroides anabólicos androgênicos (EAA), é crescente na sociedade o uso destas drogas por razões estéticas e este consumo tem aumentado principalmente entre as mulheres nas últimas décadas. É amplamente relatado que os EAA comprometem a saúde e promovem efeitos adversos na reprodução, no entanto, pouca atenção é dada a respeito dos efeitos promovidos pelos EAA no ciclo estral, na morfologia ovariana e uterina e na regulação da função ovariana após os períodos de tratamento e de recuperação. Nenhum relato foi obtido na literatura, quanto à administração de diferentes doses de EAA e a possibilidade de reversibilidade dos efeitos colaterais. Assim, o objetivo do projeto é avaliar o efeito de diferentes doses do esteroide decanoato de nandrolona (DN) no ciclo estral e nos ovários e útero de ratas albinas com ênfase no controle da imunoexpressão do AR, ERs, CYP450 aromatase e Inibina-A do tecido ovariano e nos níveis hormonais sexuais, e também se há recuperação dos prejuízos reprodutivos após a interrupção do tratamento esteroidal. Ratas Wistar foram tratadas com DN nas doses de 1,87, 3,75, 7,5 e 15 mg/kg ou óleo mineral (grupos controle) por 15 dias via subcutânea. Os animais foram divididos em três procedimentos: (a) tratamento durante 15 dias; (b) tratamento seguido por recuperação de 30 dias; (c) tratamento seguido por recuperação de 60 dias. O ciclo estral foi monitorado diariamente e no final de cada período os a... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Although are extensive the options of therapeutic use of anabolic-androgenic steroids (AAS), the use of these drugs for aesthetic reasons is growing in society and this consumption has increased mainly among women in recent decades. It is widely reported that the AAS compromise the health and promote adverse effects on reproduction, however, little attention is given on the effects promoted by the AAS in the estrous cycle, in ovarian and uterine morphology and in the regulation of ovarian function after treatment periods and recovery. No report has been obtained from the literature regarding the administration of different doses of synthetic steroids and the possibility of reversibility of side effects. The objective of this project is to evaluate the effect of different doses of steroid nandrolone decanoate (ND) in the estrous cycle and ovaries and uterus of albino rats with emphasis on control of AR immunoexpression, ERs, CYP450 aromatase and inhibin-A in the ovarian tissue and sexual hormone levels, and evaluate if there is recovery of possible reproductive damages after cessation of steroid treatment. Female Wistar rats were treated with ND at doses of 1.87, 3.75, 7.5 and 15 mg/kg or received mineral oil (control groups estrus and diestrus) for 15 days subcutaneously. The animals were divided into three procedures: (a) treatment for 15 days; (b) treatment followed by recovery to 30 days; (c) treatment followed by recovery for 60 days. The estrous cycle was monitored daily... (Complete abstract click electronic access below) / Mestre

BACKGROUND: Adjudication is a standardized, objective, and often blinded mechanism designed to assess clinical events with increased accuracy. It is performed by a centralized committee of independent reviewers, who are specialized, expert physicians who have no involvement with either the treatment of study subjects or the trial sponsor. Adjudication can decrease variability and bias in study results and increase the likelihood of correct identification, assessment, and categorization of clinical events such as potential malignancies diagnosed through histopathology. Histopathology is highly variable due to the subjective nature of the assessments. THESIS: If it is the case that there are clinically significant discrepancies between local and central diagnoses and that central adjudication yields more accurate diagnoses than a local pathologist, then it should be accepted that adjudication ought to be more widely used in clinical trials to assess histopathology-related safety outcomes and endpoints. METHODS AND STATISTICS: This is a retrospective cross-sectional study assessing interrater variability between local and central diagnoses of biopsy samples in a clinical trial setting using kappa scores and percent agreement. Certified Professional Coders (CPC) and central pathologists used the International Classification of Diseases for Oncology revision 3 (ICD-O 3) to codify the local and central assessments to permit comparison. Three statistical groups (group A: the full dataset, group B: pathology sub-specialty reading groups, and group C: non-melanoma skin cancers versus all other malignancies) were assessed for interrater variability in seven separate analyses: neoplasm versus non-neoplasm (analysis 1), benign versus malignant including non-neoplasms (analysis 2.1), benign versus malignant excluding non-neoplasms (analysis 2.2), discrepancies in morphology and/or behavior including non-neoplasms (analysis 3.1), discrepancies in morphology and/or behavior excluding non-neoplasms (analysis 3.2), all discrepancies leading to differences in treatment (analysis 4.1), and all discrepancies leading to difference in treatment with round 1 matches removed (analysis 4.2). RESULTS: 602 cases comprised the dataset. Based on kappa scores, there is near perfect agreement between the central and local lab diagnoses in analyses 1, 2.1, and 2.2 in group A (all cases in the dataset). The percent agreement for these analyses is above 90%. The group A (full dataset) kappa score and percent agreement decreased to 0.59 and 68.3%, respectively, in analysis 3.1 (discrepancies in morphology and/or behavior codes, including non-neoplasms). When non-neoplasms were removed (analysis 3.2), the kappa score and percent agreement were 0.52 and 57.0%, respectively. In group C, NMSC had substantial kappa agreement in analyses 1, 2.1, and 2.2, whereas all other malignancies had near perfect kappa agreement. All percent agreements were above 88% and surpassed the minimally acceptable threshold for interrater percent agreement in healthcare (80%). Group B divided the data set into 10 sub-specialty reading groups. Kappa scores ranged from 0.66 (GYN) to 1.00 (lung) in analysis 1; the analysis 1 kappa score for lymphoma was 0.55, but this was not statistically significant. In analysis 2.1, lung and sarcoma had the highest kappa scores (1.00) and dermatology and GYN had the lowest (0.71). As in analysis 1, the kappa score for lymphoma was 0.55 but was not statistically significant. When non-neoplasms were removed from analysis 2.2, 6 of the 10 sub-groups had kappa scores of 1.00, but all 6 had sample sizes less than 10. Percent agreement ranged from 80 to 100 percent. When all cases were considered regardless of number of rounds of review (analysis 4.1), about 90% of diagnoses would have similar courses of treatment. All sub-groups except sarcoma reached the minimally acceptable agreement rate in healthcare (80%). In the remaining 33% of cases that did not have matching diagnoses in round 1 (analysis 4.2), 34% may have different courses of treatment depending on whether the local or central diagnoses was used. Mid-study updates to the charter and CPC/reviewer manuals and processing of specimens did not have a significant impact on results. CONCLUSION: Although there is little discrepancy between local and central pathologists on whether malignancies exist among samples, there is discord regarding specific diagnoses and their associated treatments. Adjudication can assist in decreasing this discordance in order to develop the most specific and accurate safety profile for a compound.

Previous studies on fishes exposed to effluent from the Key Lake uranium mill in northern Saskatchewan have demonstrated elevated lipids in young-of-the-year pike (Esox lucius), deformities in larval pike and decreased survival of fathead minnows (Pimephales promelas). The objectives of this thesis were to evaluate possible factors that could be contributing to altered bioenergetics of juvenile northern pike inhabiting lakes receiving effluent from the Key Lake operation and to examine the effects of effluent exposure on biomarkers of oxidative stress and histopathology of target organs. Although glycogen and triglycerides stores were significantly greater in pike from exposure lakes compared to the reference, triglycerides stores of juvenile pike prey items showed no overall differences among lakes. Measures of parasitism, however, were negatively correlated with pike bioenergetics thereby reflecting a possible energetic cost of parasitism on reference lake fish. The degree of infection by intestinal parasites and gill monogeneans was greatest in reference pike and intermediate in low exposure pike, whereas high exposure pike harboured no parasites. <p>Arsenic, nickel and selenium are elevated in lakes downstream of the Key Lake mill and have been shown to be associated with increased reactive oxygen species (ROS) in biological systems causing oxidative stress. The potential for oxidative stress was assessed in pike liver and kidney using several biomarkers. Overall, the concentrations of total, reduced and oxidized glutathione and the ratio of oxidized to reduced glutathione did not differ significantly among exposure and reference pike. The activity of glutathione peroxidase was greater in high exposure than reference liver whereas, contrary to predictions, lipid peroxidation was greater in reference than exposure pike tissues. <p>Histopathological evaluations revealed greater kidney and gill pathology in reference lake pike, whereas for liver, hepatocyte morphology differed among lakes without any clear signs of pathology. Trace metal analyses of muscle showed that eight elements (arsenic, cobalt, copper, iron, molybdenum, selenium, thallium, uranium) were significantly elevated in exposure pike. These results provide only limited evidence of oxidative stress in exposure pike tissues and no evidence of histopathology despite indications that metals are bioaccumulating in tissue. Overall, the results from this thesis suggest that the health and condition of juvenile northern pike living downstream of the Key Lake uranium mill may not be compromised by effluent exposure.

Previous studies on fishes exposed to effluent from the Key Lake uranium mill in northern Saskatchewan have demonstrated elevated lipids in young-of-the-year pike (Esox lucius), deformities in larval pike and decreased survival of fathead minnows (Pimephales promelas). The objectives of this thesis were to evaluate possible factors that could be contributing to altered bioenergetics of juvenile northern pike inhabiting lakes receiving effluent from the Key Lake operation and to examine the effects of effluent exposure on biomarkers of oxidative stress and histopathology of target organs. Although glycogen and triglycerides stores were significantly greater in pike from exposure lakes compared to the reference, triglycerides stores of juvenile pike prey items showed no overall differences among lakes. Measures of parasitism, however, were negatively correlated with pike bioenergetics thereby reflecting a possible energetic cost of parasitism on reference lake fish. The degree of infection by intestinal parasites and gill monogeneans was greatest in reference pike and intermediate in low exposure pike, whereas high exposure pike harboured no parasites. <p>Arsenic, nickel and selenium are elevated in lakes downstream of the Key Lake mill and have been shown to be associated with increased reactive oxygen species (ROS) in biological systems causing oxidative stress. The potential for oxidative stress was assessed in pike liver and kidney using several biomarkers. Overall, the concentrations of total, reduced and oxidized glutathione and the ratio of oxidized to reduced glutathione did not differ significantly among exposure and reference pike. The activity of glutathione peroxidase was greater in high exposure than reference liver whereas, contrary to predictions, lipid peroxidation was greater in reference than exposure pike tissues. <p>Histopathological evaluations revealed greater kidney and gill pathology in reference lake pike, whereas for liver, hepatocyte morphology differed among lakes without any clear signs of pathology. Trace metal analyses of muscle showed that eight elements (arsenic, cobalt, copper, iron, molybdenum, selenium, thallium, uranium) were significantly elevated in exposure pike. These results provide only limited evidence of oxidative stress in exposure pike tissues and no evidence of histopathology despite indications that metals are bioaccumulating in tissue. Overall, the results from this thesis suggest that the health and condition of juvenile northern pike living downstream of the Key Lake uranium mill may not be compromised by effluent exposure.