Thesis advisor: Kenneth Williams / Approximately 15% of individuals infected with Human Immunodeficiency Virus (HIV) develop a neurological condition that consists of motor dysfunction and cognitive deterioration in late stage disease that is known as the AIDS dementia complex (ADC). This condition is mirrored in rhesus macaques infected with Simian Immunodeficiency Virus (SIV), which can be more easily studied. This project analyzed different macrophage populations in rhesus macaques infected and uninfected with SIV at early and terminal stage disease. Single and double immunohistochemistry stains were performed for the known macrophage and microglial markers CD163, CD16, CD68, Mac387, HAM56, and Iba-1, as well as for the SIV-p28 viral protein. Photographs and observations of the tissue stainings demonstrated that early after infection with SIV, there is an increase in perivascular macrophages and monocytes surrounding vessels and tissue edges, and the SIV-p28 protein is already present. There is also an observed change in the morphology of the microglia to an active, ramified state. After the development of AIDS and SIVE, the increase in all of the macrophage markers and the accumulation of activated microglia are clearly visible, especially surrounding and within lesions. Furthermore, these markers can be used to categorize the encephalitic lesions as “new” or “old” based on the presence or absence of Mac387 within the cells. All lesions contained CD68+ and HAM56+ macrophages, but “new” lesions presented with a relatively high count of Mac387+ macrophages that were newly imported from the periphery, whereas “old” lesions lacked Mac387+ cells. / Thesis (BS) — Boston College, 2009. / Submitted to: Boston College. College of Arts and Sciences. / Discipline: College Honors Program. / Discipline: Biology.

by Kwan Yiu Wing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 167-187). / Abstracts in English and Chinese. / ACKNOWLEDGMENTS --- p.IV / ABSTRACT --- p.1 / Chapter CHAPTER 1. --- INTRODUCTION --- p.5 / Chapter I. --- Epidemiology of Prostate Cancer --- p.5 / Chapter II. --- The Normal Prostate - Prostatic Anatomy --- p.9 / Chapter III. --- Pathology of Prostatic Cancers --- p.12 / Chapter CHAPTER 2. --- PROSTATIC INTRAEPITHELIAL NEOPLASIA --- p.16 / Chapter I. --- Introduction --- p.16 / Chapter II. --- "Definition, Characteristics and Grading" --- p.16 / Chapter III. --- Incidence and Prevalence of PIN --- p.26 / Chapter IV. --- Evidence Linking PIN with Prostatic Carcinoma --- p.27 / Chapter V. --- Conclusion --- p.37 / Chapter CHAPTER 3. --- HISTOLOGIC BIOMARKERS --- p.39 / Chapter I. --- p53 Protein --- p.39 / Chapter II. --- Proliferating Cell Nuclear Antigen (PCNA) --- p.44 / Chapter III. --- Ki-67 Antigen --- p.48 / Chapter IV. --- Epidermal Growth Factor Receptor --- p.49 / Chapter V. --- E-Cadherin --- p.51 / Chapter VI. --- CD44 --- p.54 / Chapter VII. --- nm23 --- p.58 / Chapter CHAPTER 4. --- OBJECTIVES OF STUDY --- p.62 / Chapter CHAPTER 5. --- MATERIALS AND METHODS --- p.63 / Chapter I. --- Materials --- p.63 / Chapter II. --- Methods --- p.71 / Chapter III. --- Interpretation of Immunostaining Results --- p.78 / Chapter IV. --- Statistical Analysis --- p.82 / Chapter CHAPTER 6. --- RESULTS --- p.83 / Chapter I. --- Immunohistochemical Results for p53 Protein --- p.83 / Chapter II. --- Results of Immunostaining of PCNA --- p.90 / Chapter III. --- Immunostaining and Quantitation of Ki-67 Expression --- p.97 / Chapter IV. --- Immunohistochemical Expression of EGFr --- p.105 / Chapter V. --- E-Cadherin --- p.110 / Chapter VI. --- CD44 --- p.115 / Chapter VII. --- Expression of nm23 in Prostatic Lesions --- p.122 / Chapter VIII. --- Correlation and Association of Expressions of All Biomarkers in Prostatic Lesions --- p.128 / Chapter CHAPTER 7. --- DISCUSSION --- p.132 / Chapter I. --- p53 Protein --- p.135 / Chapter II. --- PCNA --- p.137 / Chapter III. --- Ki-67 --- p.140 / Chapter IV. --- EGFr --- p.143 / Chapter V. --- E-Cadherin --- p.146 / Chapter VI. --- CD44 --- p.148 / Chapter VII. --- nm23 --- p.151 / Chapter VIII. --- Association between Biomarkers and Prostate lesions --- p.154 / Chapter CHAPTER 8. --- SUMMARY AND CONCLUSION --- p.157 / APPENDICES --- p.164 / Chapter I. --- Table of incidence and mortality rates of prostate cancer in the United States from 1973 to 1995 by race --- p.164 / Chapter II. --- Table of leading cancer deaths in Hong Kong from 1971 to 1996 --- p.165 / Chapter III. --- Table of incidence and mortality rate caused by prostate cancer in Hong Kong --- p.166 / Chapter IV. --- Reagents --- p.167 / REFERENCES --- p.169

Orientadora: Julieta Rodini Engrácia de Moraes / Banca: Maria de Jesus Veloso Soares / Banca: Marta Maria Circhia Pinto Luppi / Banca: Aureo Evangelista Santana / Banca: Marcia Rita Fernandes Machado / Resumo: O objetivo deste estudo foi o de caracterizar lesões musculares em cães com Distrofia Muscular do golden retriever (DMGR), de diferentes idades, por análises histopatológica e imunoistoquímica. Foram utilizados vinte e cinco cães machos classificados e distribuídos em grupos de acordo com a idade: grupo I - distróficos até 1 ano; grupo II - distróficos acima de 1 ano; grupo III - controle até 1 ano; grupo IV - controle acima de 1 ano. Uma amostra de cada músculo foi fixada em solução de formol, processadas pelas técnicas usuais de inclusão em parafina, coradas com HE e TGM para análise histopatológica e processadas para a análise imunoistoquímica de linfócitos T-CD3+, antígeno MHC II e vimentina. Outras amostras foram congeladas em nitrogênio líquido e processadas pelas técnicas usuais para realização das reações imunoistoquímicas para marcação dos linfócitos T-CD4+, TCD8+ e do antígeno MHC I. Os resultados mostraram que as lesões nos músculos distróficos do grupo I foram moderadas comparativamente às do grupo II que foram severas. Nos músculos distróficos, os linfócitos T-CD3+, T-CD4+ e T-CD8+ concentravam-se nas áreas de degeneração e necrose. O número de linfócitos T-CD3+ e T-CD4+ foi significativamente maior (p < 0.05) em todos os músculos distróficos em comparação aos controles, demonstrando a participação dos linfócitos T na doença. O número de linfócitos T-CD8+ foi significativamente maior (p < 0.05) nos distróficos, exceto para os músculos sartório cranial no grupo I, diafragma e bíceps femoral no grupo II. A imunoexpressão do MHC I intensificou-se com a idade nos animais distróficos, ao contrário do MHC II que se manteve. A imunoexpressão da vimentina e do VEGF nos músculos distróficos foi discreta (escore 1) em todos os músculos avaliados. Destes resultados podemos... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The purpose of this study was to characterize the lesions in dystrophic muscles of DMGR dogs of different ages, by means of histopathological and immunohistochemistry analysis. Twenty-five male dogs were classified and distributed into groups according to the age: Group I - dystrophic up to 1 year, group II - dystrophic over 1 year, group III - control up to 1 year, group IV - control over 1 year. One sample from each muscle was fixed in formalin solution, processed by usual techniques of paraffin embedding, stained with HE and TGM for histopathological purposes and processed for immunohistochemical analysis of the distribution of T-lymphocytes CD3+, MHC II and vimentin. Other samples were frozen in liquid nitrogen, processed by usual techniques of freezing in order to perform the techniques of immunohistochemical labelling for CD4+ T-lymphocytes, T-CD8+ and MHC I. The results of histopathological analysis showed that the lesions in dystrophic muscles in the Group I were moderate compared to that ones in the Group II which were severe. The CD3+, CD4+ and CD8+ Tlymphocytes were more numerous in dystrophic muscles especially in areas of degeneration and necrosis. The number of CD3+ and CD4+ T-lymphocytes was found to be significantly higher (p <0.05) in all dystrophic muscles compared to controls demonstrating the involvement of T-lymphocytes in the disease. The number of CD8+ Tlymphocytes was found to be significantly higher (p <0.05) in dystrophics, except for the cranial sartorius muscles in the Group I and the diaphragm and biceps femoris in the Group II. The immunoexpression of MHC I increased with age in dystrophic animals, in contrast to MHC II that remained the same. The immunoexpression of vimentin and VEGF in the dystrophic muscles was mild (score 1) upon all muscles. From these results we can conclude that in dystrophic muscle immunoexpression of... (Complete abstract click electronic access below) / Doutor

Orientador: Luiz Cesar de Moraes / Banca: João Luiz de Miranda / Banca: Luiz Cesar de Moraes / Banca: Miguel Angel Castilho Salgado / Banca: Yasmin Rodarte Carvalho / Banca: Marlene Fenyo Soeiro de Matos Pereira / Resumo: Pacientes portadores de câncer na região de cabeça e pescoço quando submetidos à radioterapia podem apresentar vários tipos de manifestações clínicas, dentre elas a diminuição dos níveis salivares do fator de crescimento epidérmico (EGF). O EGF é uma pequena proteína (53 aminoácidos) que estimula a proliferação de células dos mamíferos, sendo encontrada em vários órgãos em desenvolvimento. Pode também exercer um papel fisiológico na erupção dentária ao interagir com outras moléculas como o fator de crescimento transformante β (TGF-β), a interleucina 1 (IL-1) e do fator de estimulação de colônia 1 (CSF-1), aumentando a reabsorção óssea e estimulando a quimiotaxia de células mononucleares. O objetivo deste trabalho foi verificar, por meio de reações de imuno-histoquímica, se a expressão do fator de crescimento epidérmico (EGF) pode ser alterada na odontogênese do primeiro molar superior de camundongos Mus musculus, após exposição de fêmeas prenhes a radioterapia, na dose de 3 Gray (Gy) ao décimo dia de gestação. Foram avaliados os germes dentários dos embriões aos 14, 16 e 18 dias de desenvolvimento pré-natal. As análises morfológica microscópica óptica e histomorfométrica demonstraram que o número de células epiteliais periféricas do órgão do esmalte imunopositivas para o EGF foi significativamente menor no grupo 3 Gy em relação ao grupo controle nos períodos de 14o (P<0,0001), 16o (P<0,0001<0,05) e 18o (P<0,0008) dias pré-natais. Processo FAPESP no 2008/54534-8. / Abstract: Malignant neoplasm orofacial patients when receiving radiation therapy can present several types of radiation injuries and clinical manifestations, such as decrease of the salivary epidermal growth factor (EGF) levels. EGF is a small protein (53 amino acids) that stimulates the proliferation of cells of the mammals, being found in several organs in development. EGF can exercise a physiological role in the dental eruption through the interaction with other molecules such as transforming growth factor beta (TGF-β), interleukin-1 (IL-1) and colony-stimulating factor-1 (CSF-1), which increases the bone reabsorption and stimulating the chemotaxis for mononuclear cells. The objective of this work was to verify through technique of immunohistochemistry, changes in the expression of the EGF in the odontogenesis of the first upper molar in Mus musculus mice embryos to the 14th, 16th and 18th days of intrauterine life. Pregnant mice was irradiated on the 10th gestacional day with a 3 Gray (Gy) dose. The microscopical mophological and histomorphometrical analysis showed a significant decrease in the number of EGF-positive dental epithelium cells in the 3 Gy group when compared with control group in 14th (P<0,0001), 16th (P<0,0001<0,05) and 18th (P<0,0008) intrauterine day periods. Supported by FAPESP no 2008/54534-8. / Doutor

According to the UNAIDS global report 2013, an estimated 6.1 million people are living with human immunodeficiency virus (HIV) in South Africa. The incidence of Kaposi sarcoma (KS) has increased dramatically since the Acquired Immunodeficiency Syndrome (AIDS) epidemic. Of the estimated 66 200 cases of KS worldwide, 58 800 are thought to have occurred in SSA (Parkin 2002). However, there remains a paucity of published data about KS from South Africa. This retrospective study was conducted to describe the epidemiology of KS at Chris Hani Baragwanath Academic Hospital (CHBAH) and to determine possible links among the CD4 counts, intensity and distribution of human herpes virus 8 latency- associated nuclear antigen 1 (HHV8 LNA-1) immunohistochemical staining and the stage of KS. Nine hundred and thirty eight histopathology reports of KS diagnosed in 901 patients at CHBAH between 2005 and 2009 were reviewed and demographic data (age, gender, topographic site, CD4 count, HIV status, KS stage, HHV8 LNA-1 staining, concomitant pathology) were recorded. The H&E stained sections and HHV8 LNA-1 immunostains of a cohort of 127 cases were subsequently reviewed and categorised with regard to intensity and distribution of staining. The male:female ratio was 1,2:1. The mean age was 36,8 years (standard deviation {SD} 10,2 years) and the median CD4 count 127,5 cells/mm3 (quartile range {QR} 184,5 cells/mm3). Lower limb skin biopsies accounted for 49,6% of cases. Concomitant pathology was seen in 4,6% of cases. Infections and inflammatory dermatoses were the most frequently diagnosed concomitant pathology in cutaneous biopsies. Paediatric, visceral and endemic KS accounted for only limited proportions of cases (1,44% of patients; 1,4% and 1,3% respectively). There was a significant difference in the distribution of HHV8 LNA-1 staining in patch versus nodular KS (p = 0,011). The CD4 counts were not predictive of KS Page | v stage (p = 0,701) or the intensity (p = 0,877) and distribution (p = 0,846) of HHV8 LNA-1 immunohistochemical staining. This study highlights the epidemiology of KS and the variability in HHV8 LNA-1 immunohistochemical staining across CD4 counts and stages of KS.

Abstract not available

<p>This study was carried out to determine the phenotype of special dog mammary gland tumours that were grown in nude mice. 26 tumours were examined by the immunohistochemical ABC-Elite protocol. The tumour tissues were labelled with following anti-human antibodies:</p><p>- AE1/AE3 (pankeratin antibody) labelled epithelial and myoepithelial cells</p><p>- CD 31 labelled endothelial cells</p><p>- desmin labelled cross-striated and smooth muscle cells</p><p>- myosin labelled cross striated muscle cells</p><p>- neurofilament (NF) labelled nerve cells</p><p>- osteopontin labelled preosteoblasts, osteoblasts and osteocytes</p><p>- p63 labelled nuclei of the myoepithelial cells</p><p>- smooth muscle actin (SMA) labelled the cytoplasm of myoepithelial cells</p><p>- type I collagen labelled the extracellular matrix in connective tissue and bone</p><p>- type II collagen labelled the extracellular matrix in cartilage</p><p>- vimentin labelled fibroblasts, fibrocytes, lipocytes, smooth muscle cells, endothelial cells, nerve cells, macrophages and myoepithelial cells</p><p>The tumours were also submitted to a double immunolabelling study using p63 and SMA.</p><p>The study could not give a final conclusion about the origin the tumours. There was still need for more research to answer that question. However, the immunohistochemical technique was analysed in detail, in order to obtain perfect labelings.</p><p>Initially, all the antibodies were tested on normal dog tissue, to acquire the best working dilutions with the lowest background problems. In the tumours, good results were obtained with these dilutions for the antibodies p63, SMA, vimentin, desmin, NF, AE1/AE3 and CD 31. Except for type I collagen, type II collagen and osteopontin that gave too much unspecific labelling of the mouse connective tissue. Even, when using the Vector® M.O.M. blocking kit, the results were still very difficult to interpretate.</p><p>The antigen retrieval methods were evaluated for all the antibodies. The antibodies p63, SMA, vimentin, desmin, AE1/AE3, myosin, neurofilament and CD 31 needed the antigen retrieval treatment. The antibodies type I collagen and type II collagen needed the treatment with the enzyme pepsin, while osteopontin did not need any pretreatment at all.</p><p>The double immunolabelling with p63 and SMA gave excellent results. Different combinations were tried out with different substrates, namely Vector® Nova RED, Vector® DAB and Vector® SG. Vector® methyl green was used as counterstaining, but it interfered with the other substrates, and better results were obtained without this counterstaining.</p>

The blood-brain barrier regulates the transport into the brain and protects the central nerve system (CNS) from toxics substances. However some areas of the brain, called circumventricular organs (CVO), lack the blood-brain barrier. One of these is area postrema (AP), which is located in the brainstem immediately adjacent to the nucleus tractus solitarius (NTS). These two areas together regulate autonomic behaviours such as food intake, and also make up the vomiting center. The hormones leptin and ghrelin, which regulate food intake, are too big to pass the blood-brain barrier, but have receptors in NTS. In this study we used immunohistochemistry to obtain a detailed map of the different components of the blood-brain barrier in AP and NTS. The results suggest that there is a barrier that prevents diffusion of substances from AP into NTS. However, there seems to be some vessels in NTS that have a weaker or no barrier characteristics. These vessels could provide an entrance for peripheral substances to neurons in NTS.

Bovine cysticercosis is a zoonotic disease for which cattle are the intermediate hosts of the human tapeworm <i>Taenia saginata</i>. Routine inspection measures are implemented in Canada by the Canadian Food Inspection Agency (CFIA), and similarly elsewhere, for the postmortem detection of larval parasite cysts (cysticerci) in beef destined for human consumption. Detection is based on the gross examination of traditional carcass predilection sites, although it is recognized that the parasite has no true predilection for a particular tissue or site. In order to evaluate the efficacy of the inspection protocol currently implemented in Canada, a study was undertaken to determine the distribution of <i>T. saginata</i> cysticerci in tissues of experimentally infected cattle. Forty-two cross-bred beef cattle were divided into five groups of 5-12 animals each and inoculated orally with either 10000, 5000, 1000, 100 or 10 <i>T. saginata</i> eggs obtained from cases of human taeniosis in Thailand. From 47 to 376 days post-inoculation (DPI), ten animals inoculated with 5000 eggs were killed and the carcasses partitioned into 31 tissue sites. These consisted of the traditionally inspected tissue sites of heart, masseter and pterygoid muscles, tongue, oesophagus, and diaphragm (membranous and crura); as well as non-traditional sites of lung, liver and 20 additional muscles or muscle groups. After the routine inspection for cysticerci of traditional tissue sites, tissues from all sites were each cut into approximately 0.5 cm thick slices and the total number of parasitic cysts and cyst density (cysts/g of tissue) were determined for each site. Traditional sites were similarly evaluated for the remaining 32 animals that were killed between 117 and 466 DPI. Sites were ranked based on cyst density. In the animals for which non-traditional sites were also evaluated, no sites had higher cyst densities than those traditionally inspected. When only traditional sites for all animals were compared, the heart ranked highest overall, although not significantly different from masseter, and was the most frequently affected site. The traditional site of oesophagus was among the poorest of all sites for detection of cysticerci. The heart was confirmed as the site of choice for detection of bovine cysticercosis based on high cyst density and frequency of infection. There was also enhanced visibility of parasite lesions in the heart due to the relatively early degeneration and resultant gross pathology that occurs in cardiac muscle. More thorough examination of the heart is recommended during post-mortem inspection for this parasite, particularly when examining animals from an infected herd. <p>Currently, confirmation by CFIA of suspect cysticerci recovered during meat inspection relies on gross, stereomicroscopic, or standard histological examination. Although degenerating cysticerci are more likely to be detected and submitted for diagnosis, they often cannot be definitively identified by these methods. A recently developed monoclonal antibody-based immunohistochemical (IHC) assay for post-mortem diagnosis of this parasite was optimized and standardized. The IHC method was compared to the currently used histological assay using 169 degenerated known-positive <i>T. saginata</i> cysticerci collected from the experimental infections in the first study and from field submissions, and known-negative specimens and lesions of various etiologies from non-infected cattle. The use of the IHC assay identified significantly more known-positive bovine cysticerci (91.7%) than the histological method (38.5%), and non-specifically reacted only with the other cestode species examined. Since <i>T. saginata</i> is the only larval cestode typically found in the muscle of cattle, this cross-reactivity is not significant and the IHC assay will be a useful tool for the identification of lesions caused by degenerated bovine cysticerci.<p>This research provided evidence to support changes to the current post-mortem inspection, detection and diagnostic procedures and will contribute to more effective and efficient control of bovine cysticercosis.

Bovine cysticercosis is a zoonotic disease for which cattle are the intermediate hosts of the human tapeworm <i>Taenia saginata</i>. Routine inspection measures are implemented in Canada by the Canadian Food Inspection Agency (CFIA), and similarly elsewhere, for the postmortem detection of larval parasite cysts (cysticerci) in beef destined for human consumption. Detection is based on the gross examination of traditional carcass predilection sites, although it is recognized that the parasite has no true predilection for a particular tissue or site. In order to evaluate the efficacy of the inspection protocol currently implemented in Canada, a study was undertaken to determine the distribution of <i>T. saginata</i> cysticerci in tissues of experimentally infected cattle. Forty-two cross-bred beef cattle were divided into five groups of 5-12 animals each and inoculated orally with either 10000, 5000, 1000, 100 or 10 <i>T. saginata</i> eggs obtained from cases of human taeniosis in Thailand. From 47 to 376 days post-inoculation (DPI), ten animals inoculated with 5000 eggs were killed and the carcasses partitioned into 31 tissue sites. These consisted of the traditionally inspected tissue sites of heart, masseter and pterygoid muscles, tongue, oesophagus, and diaphragm (membranous and crura); as well as non-traditional sites of lung, liver and 20 additional muscles or muscle groups. After the routine inspection for cysticerci of traditional tissue sites, tissues from all sites were each cut into approximately 0.5 cm thick slices and the total number of parasitic cysts and cyst density (cysts/g of tissue) were determined for each site. Traditional sites were similarly evaluated for the remaining 32 animals that were killed between 117 and 466 DPI. Sites were ranked based on cyst density. In the animals for which non-traditional sites were also evaluated, no sites had higher cyst densities than those traditionally inspected. When only traditional sites for all animals were compared, the heart ranked highest overall, although not significantly different from masseter, and was the most frequently affected site. The traditional site of oesophagus was among the poorest of all sites for detection of cysticerci. The heart was confirmed as the site of choice for detection of bovine cysticercosis based on high cyst density and frequency of infection. There was also enhanced visibility of parasite lesions in the heart due to the relatively early degeneration and resultant gross pathology that occurs in cardiac muscle. More thorough examination of the heart is recommended during post-mortem inspection for this parasite, particularly when examining animals from an infected herd. <p>Currently, confirmation by CFIA of suspect cysticerci recovered during meat inspection relies on gross, stereomicroscopic, or standard histological examination. Although degenerating cysticerci are more likely to be detected and submitted for diagnosis, they often cannot be definitively identified by these methods. A recently developed monoclonal antibody-based immunohistochemical (IHC) assay for post-mortem diagnosis of this parasite was optimized and standardized. The IHC method was compared to the currently used histological assay using 169 degenerated known-positive <i>T. saginata</i> cysticerci collected from the experimental infections in the first study and from field submissions, and known-negative specimens and lesions of various etiologies from non-infected cattle. The use of the IHC assay identified significantly more known-positive bovine cysticerci (91.7%) than the histological method (38.5%), and non-specifically reacted only with the other cestode species examined. Since <i>T. saginata</i> is the only larval cestode typically found in the muscle of cattle, this cross-reactivity is not significant and the IHC assay will be a useful tool for the identification of lesions caused by degenerated bovine cysticerci.<p>This research provided evidence to support changes to the current post-mortem inspection, detection and diagnostic procedures and will contribute to more effective and efficient control of bovine cysticercosis.