Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-02-15T15:46:57Z No. of bitstreams: 1 2015 - Ana?s Gabriela Villarreal Laguna.pdf: 1531103 bytes, checksum: 8567bc787b67e1d16f71708035dbfc97 (MD5) / Made available in DSpace on 2017-02-15T15:46:58Z (GMT). No. of bitstreams: 1 2015 - Ana?s Gabriela Villarreal Laguna.pdf: 1531103 bytes, checksum: 8567bc787b67e1d16f71708035dbfc97 (MD5) Previous issue date: 2015-12-21 / Otitis is defined as an inflammation, acute or chronic, of the ear canal and it?s considered one of the most common diseases in small animal practice. It possesses various perpetuating agents, bacteria and yeast being the most common. Treatment is based in the recognition of the primary causes and predisposing factors, and the resolution of secondary infections. A greater sensitivity in feline ear canal has also been shown, when compared to dogs. With this in mind, a product with the least possible active ingredients would be the ideal topical treatment in felines. However, the majority of available products combine antibiotics and antifungals, this combination not being necessary in all cases; for bacterial otitis, very few specific products exist, and some of them present a proven ototoxic potential. In face of this problem, the efficacy of a new formulation using levofloxacin for the specific treatment of bacterial otitis was made by the Farmacologia e Farmacometria sector of the LQEPV ? UFRRJ, using isopropylic alcohol as vehicle. Eighteen adult felines, belonging to the Laborat?rio de Quimioterapia Experimental em Parasitologia Veterin?ria (LQEPV-UFRRJ), diagnosed with bacterial otitis according to clinical signs (erythema, inflammation, pruritus, hyperpigmentation and secretions), as well as cytological exams and video-otoscopy, were chosen. Individual data was registered in specific formularies for comparison of pretreatment and post treatments evaluations. After the initial evaluation the animals were divided into three groups, according to treatment: in Group A or control (6 animals) a placebo (vehicle) was used; in Group B (6 animals) a formulation of levofloxacin 0, 5% was used; Group C (6 animals) a formulation of de levofloxacin 0, 5% + dexamethasone 0, 02% was used. In every case 0, 5 ml was applied to each ear, once a day for 15 consecutive days. Post treatment video-otoscopies were performed on day +21, and post treatment cytologies were performed on days +7, +14 and +21. The animals were always observed by the same evaluator, avoiding errors of concordance and checking for possible reactions to the product. None of the animals presented adverse reactions to the product during the treatment period, besides slight discomfort immediately after application. Animals from Group A continued with the same scores until the end of the experiment. Animals from Group B presented an initial clinical improvement, but some of them developed a fungal otitis as time passed. Animals from Group C presented a significant improvement since day +7, with no secondary alterations through time. It is demonstrated that a formulation of de levofloxacin + dexamethasone can be efficient and safe for the treatment of feline bacterial otitis / A otite ? definida como uma inflama??o, aguda ou cr?nica, do conduto auditivo e ? uma das doen?as mais frequentes na cl?nica de animais de companhia. Possui diversos agentes perpetuantes, sendo mais comuns as bact?rias e leveduras. O tratamento e baseado no reconhecimento de causas principais e fatores predisponentes, assim como a resolu??o de infec??es secund?rias. Tem sido relata uma sensibilidade maior nos condutos auditivos dos gatos, quando comparados com c?es. Deste modo, um produto com poucos ingredientes farmacologicamente ativos seria o tratamento t?pico ideal em felinos. No entanto, a maioria dos produtos para otite dispon?veis no mercado combina tanto antibi?ticos, como antif?ngicos, n?o sendo esta combina??o necess?ria em todos os casos; especificamente para otites bacterianas, existem poucos produtos espec?ficos, e muitos apresentam um potencial otot?xico j? comprovado. Perante este problema, foi avaliada a efic?cia de uma nova formula??o a utilizando levofloxacina para o tratamento espec?fico da otite bacteriana em felinos, o produto foi confeccionado pelo setor de Farmacologia e Farmacometria do LQEPV ? UFRRJ, utilizando como ve?culo ?lcool isoprop?lico. Foram selecionados 18 felinos adultos pertencentes ao Laborat?rio de Quimioterapia Experimental em Parasitologia Veterin?ria (LQEPV-UFRRJ), diagnosticados com otite bacteriana de acordo com sinais cl?nicos (eritema, inflama??o, prurido, hiperpigmenta??o e presen?a de secre??o), exame citol?gico e v?deo-otoscopia. Os dados individuais foram registrados em formul?rios espec?ficos para compara??o das avalia??es inicias e p?s-tratamento. Ap?s as avalia??es iniciais, os animais foram divididos em tr?s grupos experimentais: no grupo A, controle, seis animais utilizaram um Placebo (ve?culo); no grupo B, seis animais utilizaram a formula??o contendo Levofloxacina a 0,5%; e no grupo C, seis animais foram medicados com a associa??o de Levofloxacina 0,5% com dexametasona 0,02%. Em todos os casos foram instilados 0,5ml em ambas as orelhas uma vez por dia, durante 15 dias consecutivos. Exames vide-otosc?picos p?s-tratamento foram realizadas no dia +21, e citologias nos dias +7, +14 e +21. Os animais foram observados diariamente sempre pelo mesmo avaliador, evitando assim erros de concord?ncia e acompanhando poss?veis rea??es adversas ao produto. Nenhum animal apresentou rea??o adversa ao produto durante o per?odo de tratamento, al?m de incomodo no momento imediato ap?s a aplica??o. Os animais do grupo A continuaram com o mesmo score registrado no dia 0 at? finalizado o experimento. Os animais do grupo B apresentaram melhoria inicial, porem alguns deles desenvolveram otite f?ngica ao decorrer do tempo. J? os animais do grupo C apresentaram uma melhora significativa desde o dia +7, sem altera??es secund?rias no decorrer do experimento. Demostra-se assim que a associa??o de levofloxacina 0,5% com dexametasona 0,02% pode ser eficaz e segura para o tratamento da otite bacteriana em felinos

A intensificação da criação de felinos em todo o mundo demandam uma ampliação e aprofundamento dos conhecimentos da clínica e patologia de doenças infecciosas que acometem os gatos. Entretanto, existe uma escassez de informações sobre doenças infecciosas que acometem os gatos do Brasil, especialmente da região sul. Esta tese de doutorado teve como objetivo realizar um levantamento das principais causas de morte em gatos, além da caracterização anatomopatológica e identificação de agentes infecciosos em tecidos e células de gatos através da técnica de imuno-histoquímica (IHQ) e imunocitoquímica (ICQ). O primeiro artigo teve como objetivo determinar as principais causas de morte em gatos na região Sul do Brasil, através de um estudo retrospectivo, de 2000 a 2015. Foram realizadas um total de 1753 necropsias de felinos domésticos, destas 1364 (77,8%) foram conclusivas e 389 foram inconclusivos (22,2%). As categorias mais prevalentes foram: neoplasmas (20%), doenças infecciosas/parasitárias (17,8%), doenças do sistema digestório (14,1%), traumatismos (13,4%), doenças do sistema urinário (11,4%), doenças do sistema cardiovascular (6,9%), doenças do sistema respiratório (5,8%) e outras (8,5%). As causas de morte mais frequentes incluíram: politraumatismo (13,4%), linfoma (8,8%) e peritonite infecciosa felina (7,9%). O segundo artigo teve como objetivo fazer a identificação por ICQ dos antígenos do coronavírus felino (FCoV) em macrófagos e monócitos fixados em formol e embebidos em parafina a partir de derrame e sangue como alternativa diagnóstica ante mortem da peritonite infecciosa felina. Foram selecionados para o estudo gatos com pelo menos um dos seguintes sinais clínicos: derrame em cavidades abdominal e/ou torácica e/ou pericárdica, temperatura retal acima de 40oC, icterícia, linfoadenomegalia, e sinais clínicos oftálmicos e neurológicos. Foram recebidas 25 amostras provenientes de derrames. Dessas, 16 tiveram marcação positiva no interior de macrófagos. Nove desses casos, foram confirmados pela necropsia. De 17 amostras de sangue total, 3 amostras tiveram marcação positiva no interior de monócitos; dois desses casos foram confirmadas na necropsia. O teste de ICQ anti-FCoV foi sensível e específico como método diagnóstico ante mortem da PIF. O terceiro artigo teve como objetivo descrever cinco casos de gatos apresentando cardiomiopatia hipertrófica e miocardite associadas ao vírus da imunodeficiência felina (FIV). Na necropsia, os cinco gatos apresentaram o coração acentuadamente aumentado de tamanho, por hipertrofia ventricular esquerda, e múltiplos focos brancacentos coalescentes no miocárdio e no epicárdico. Microscopicamente, no miocárdio, havia infiltrado multifocal acentuado composto por linfócitos, alguns macrófagos, neutrófilos e plasmócitos. Na IHQ para FIV houve intensa imunomarcação no citoplasma e no núcleo, principalmente de linfócitos e no citoplasma de ocasionais macrófagos no miocárdio. O infiltrado inflamatório caracterizou-se por linfócitos T e macrófagos, o que foi evidenciado por imunomarcação específica para essas células. Este trabalho demonstra a importância das doenças infecciosas/parasitárias, consideradas como causa de morte em gatos, demonstra a utilidade da técnica técnica ocorrência de infecção pelo do vírus da imunodeficiência felina em células inflamatórias no miocárdio de gatos com miocardite e cardiomiopatia hipertrófica. / The increase of cat breeding worldwide demands an expansion in qualified information on clinic clinical and pathological aspects of feline infectious diseases. There is, however, a lack of information on the infectious diseases that affect cats in Brazil, especially in the Southern region. This doctoral thesis aimed to survey of the main causes of death caused in cats, as well as the anatomopathological characterization of tissue response and identification of infectious agents in the tissues and cells through immunohistochemistry (IHC) and immunocytochemistry (ICH). Three articles were written. The first one aimed to determine the main causes of death in cats in southern Brazil, through a retrospective study, from 2000 to 2015. A total of 1,753 domestic feline necropsies were reviewed, from which 1,364 (77.8%) had a conclusive diagnosis and 389 were inconclusive (22.2%). The most prevalent categories were neoplasms (20%), infectious/parasitic diseases (17.8%), diseases of the digestive system (14.1%), traumatisms (13.4%), diseases of the urinary system (6.9%), diseases of the respiratory system (5.8%) and others (8.5%). The most frequent causes of death included multiple trauma (13.4%), lymphoma (8.8%) and feline infectious peritonitis (7.9%). lymphoma (8.8%) and feline infectious peritonitis (7.9%). The second article aimed to tests and alternative antemortem diagnostic method to identified cats with feline infectious peritonitis feline coronavirus (FCoV) antigens; the method consists of applying ICH to macrophages and monocytes sampled from effusion and blood, fixed in formalin and embedded in paraffin. Cats with at least one of the following clinical signs were selected for being tested: cavitary (abdominal and/or thoracic and/or pericardial) effusion, rectal temperature above 40 ° C, jaundice, enlargement of lymph nodes, clinical ophthalmic and neurological signs. Twenty-five samples from effusions were received, of which 16 had positive marking within macrophages, 9 of which were confirmed by necropsy examination. Seventeen samples consisted of whole blood, from these, 3 samples had positive staining within monocytes and two cases were confirmed by postmortem examination. The anti-FCoV ICQ test was sensitive and specific as a PIF antemortem diagnostic method. The second article aimed to identify by ICQ the feline coronavirus (FCoV) antigens in macrophages and monocytes fixed in formalin and embedded in paraffin from effusion and blood as a diagnostic alternative antemortem of feline infectious peritonitis. Cats with at least one of the following clinical signs were selected: cavitary (abdominal and/or thoracic and/or pericardial) effusion, rectal temperature above 40°C, jaundice, enlargement of lymph nodes, clinical ophthalmic and neurological signs. Twenty-five samples from effusions were received, of which 16 had positive marking inside macrophages, 9 of which were confirmed by necropsy examination. 17 samples were also received from whole blood, these 3 samples had positive staining within monocytes and in 2 cases confirmed by postmortem examination. The anti-FCoV ICQ test was sensitive and specific as a PIF antemortem diagnostic method. The third article aimed to describe five cases of cats presenting hypertrophic cardiomyopathy and myocarditis associated with feline immunodeficiency virus (FIV). At necropsy, the five cats presented a markedly enlarged heart, marked left ventricular hypertrophy, multiple myocardial and epicardial whitening foci. Microscopically, in the myocardium, there was marked multifocal infiltrate composed of lymphocytes, macrophages, neutrophils and plasma cells. In the IHC for FIV there was intense immunostaining in the cytoplasm and nucleus, mainly of lymphocytes and occasional macrophages cytoplasm in the myocardium. The inflammatory infiltrate was predominantly composed of T lymphocytes and macrophages, evidenced by immunostaining. This work demonstrates the importance of 8 infectious/parasitic diseases, as the cause of death in cats, demonstrate the use of the IHC technique as an alternative antemortem diagnostic tool for FIP and the occurrence of feline immunodeficiency virus infection in inflammatory cells in the myocardium of cats with myocarditis and hypertrophic cardiomyopathy.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-05-03T19:48:33Z No. of bitstreams: 1 2016 - Lara Patricia Santos Carrasco.pdf: 1457762 bytes, checksum: 82c38851cf4f7b3e95d40cfbb6f7ddee (MD5) / Made available in DSpace on 2018-05-03T19:48:33Z (GMT). No. of bitstreams: 1 2016 - Lara Patricia Santos Carrasco.pdf: 1457762 bytes, checksum: 82c38851cf4f7b3e95d40cfbb6f7ddee (MD5) Previous issue date: 2016-02-16 / Tritrichomonas foetus has been recently recognized as an important cause of diarrhea in various countries. The aim of this study was to diagnose and treat the infection by T. foetus in cats treated at HVPA of UFRRJ, identify the clinical relevance of this parasite, the risk factors, presence of co-infections and clinical response to treatment. In this study, were admitted 100 felines, from different breeds, both sexes and from 16 weeks of age. As inclusion criteria, patients had to have diarrhea history for more than three weeks, with little success of clinical response to treatments previously used. From the stool samples obtained by colon washing or spontaneous defecation it was conducted direct examination of stool, centrifugal flotation technique and real-time PCR. Of the 100 cats selected for this study, 10% had T. foetus infection. Most positive animals were over one year old (70%) and only 30% had up to 1 year old. Cats were 80% purebred and 20% were animals mongrel. The clinical signs present were diarrhea (60%), weight loss (10%), hemoatochezia (30%) and presence of mucus in the stool (20%). Only two animals were positive for the antigen of FeLV virus, and theses showed more severe case of diarrhea. Three animals showed co-infections with other enteropathogens (30%), one with Giardia, one with Cryptosporidium and one with Cystoisospora. All treated animals showed clinical improvement with the administrations of ronidazole at a dose 30mg/kg every 24 hours during 14 days and showed no recurrence until the recent time. We can conclude that T. foetus infection is an important differential diagnosis in cats with chronic diarrhea, which cats of any age and regardless of whether purebred or not, may have the infection, other enteropathogens needs to be investigated in all cases and the treatment was effective in the resolution of clinical signs of all treated cats. / Tritrichomonas foetus foi recentemente reconhecido como um importante agente etiol?gico de diarreia em diversos pa?ses. O objetivo deste estudo foi diagnosticar e tratar a infec??o por T. foetus nos gatos atendidos no HVPA da UFRRJ, identificar a relev?ncia cl?nica deste parasita, os fatores de risco, a presen?a de co- infec??es e resposta cl?nica ao tratamento. Neste trabalho, foram admitidos 100 animais da esp?cie felina, de diferentes ra?as, de ambos os sexos e a partir de 16 semanas de idade. Como crit?rio de inclus?o, os pacientes deveriam apresentar hist?rico de diarreia a mais de tr?s semanas, com pouco sucesso de resposta cl?nica para terap?uticas previamente institu?das. A partir das amostras de fezes obtidas por coleta via lavagem do colon dos animais e pelo ato de defeca??o espont?nea, foram realizados os m?todos do exame direto das fezes, t?cnica de centrifugo flutua??o e PCR em tempo real. Dos 100 gatos que foram selecionados para esta pesquisa, 10% apresentaram infec??o por T. foetus. A maioria dos animais positivos estavam na faixa et?ria acima de um ano (70%) e apenas 30% tinha at? um ano de idade. Os gatos eram 80% de ra?a pura e 20% eram animais sem ra?a definida. Os sinais cl?nicos apresentados por esses gatos foram diarreia (60%), perda de peso (10%), hematoquezia (30%) e presen?a de muco nas fezes (20%). Apenas dois animais infectados eram positivos para o ant?geno do v?rus da FeLV, e estes apresentaram um quadro mais severo de diarreia. Tr?s animais apresentaram coinfec??es com outros enteropat?genos (30%), um com Giardia sp, um com Cryptosporidium e outro com Cystoisospora. Todos os gatos tratados apresentaram melhora cl?nica com a administra??o do f?rmaco ronidazol na dose de 30 mg/Kg a cada 24 horas por 14 dias, n?o apresentando recidiva da diarreia at? o recente momento. Podemos concluir que a infec??o por T. foetus ? um importante diagn?stico diferencial em felinos com diarreia cr?nica, que gatos de qualquer idade e independente de ser de ra?a pura ou n?o, podem apresentar a infec??o, que a co- infec??o com outros enteropat?genos necessita ser investigada em todos os casos e que o tratamento foi efetivo na resolu??o dos sinais cl?nicos de todos os gatos tratados.

Doença renal crônica é uma afecção comum em cães e gatos, especialmente em gatos senis, e importante causa de morbidade e mortalidade. Além da idade avançada, fatores como alterações congênitas, alimentação inadequada, utilização de drogas nefrotóxicas, intoxicações e doenças infecciosas predispõem às lesões do parênquima renal. Em felinos a descrição histopatológica mais frequente é a nefrite tubulointersticial difusa. Independente da causa do dano ao néfron, a doença renal crônica é uma afecção irreversível e geralmente progressiva. Este estudo teve por objetivo realizar uma análise da contribuição da ultrassonografia utilizando-se o modo B e Doppler colorido e pulsado para o diagnóstico e estagiamento da doença renal crônica em 45 felinos. Sendo 16 do grupo controle, felinos que não portavam manifestações clínicas relacionadas ao sistema urinário, apresentavam concentração sérica de creatinina inferior a 1,6mg/dL; quatro do estágio 1, felinos com taxas de creatinina sérica inferior a 1,6mg/dL com alterações ultrassonográficas; 17 do estágio II, felinos com taxas de creatinina 1,6 a 2,8mg/dL; oito do estágio III / IV, felinos com taxa de creatinina sérica de 2,9 a 5,0mg/dL agrupados aos felinos com taxa de creatinina sérica acima de 5,0mg/dL. Os rins foram avaliados ultrassonograficamente por meio das características: ecogenicidade da cortical, regularidade de contorno e definição corticomedular no modo B; comprimento, largura e altura nos cortes longitudinal, transversal e dorsal no modo B; preenchimento das artérias interlobares, arqueadas e interlobulares por meio do Doppler colorido; e índice de resistividade dos vasos intrarenais por meio do Doppler pulsado. Foi determinada a relação comprimento do rim pelo diâmetro luminal da aorta nos felinos normais e nefropatas. Conclui-se que o aumento da ecogenicidade da cortical demonstrou-se uma característica relevante a ser considerada na avaliação ultrassonográfica da doença renal crônica. O Doppler colorido mostrou-se uma ferramenta importante no diagnóstico da doença renal crônica, especialmente quando as alterações detectadas à avaliação ultrassonográfica pelo modo B não eram expressivas. O índice de resistividade não se apresentou acima do limite da normalidade nos estágios iniciais da doença renal crônica, sugerindo não ser útil como preditor da nefropatia crônica. As alterações na avaliação ultrassonográfica modo B associadas ao aumento do índice de resistividade podem indicar um pior prognóstico da evolução da doença renal crônica. Achados ultrassonográficos como ecogenicidade da cortical aumentada, irregularidade de contorno, indefinição corticomedular, diminuição do preenchimento vascular pelo Doppler colorido e aumento do índice de resistividade são elementos importantes a serem considerados no estabelecimento do diagnostico da doença renal crônica em felinos. / Chronic kidney disease is common in dogs and cats, especially in older cats, and an important cause of morbidity and mortality. Factors like advanced age, congenital alterations, inappropriate nutrition, use of nephrotoxic drugs, intoxications and infectious disease may lead to parenchymal lesions in kidney. The most frequent histopathologic change in cats is diffuse tubulointertitial nephritis. Independent of the cause of the nephron damage, the chronic renal disease is an irreversible and usually progressive affection. This study aim to analyze the contribution of ultrasography in the diagnosis and staging of chronic kidney diseases using B mode, color Doppler and spectral Doppler. Sixteen presenting no manifestation of urinary disease and serum creatinine levels less than 1,6mg/dL served at control group; four cats represented stage I, with serum creatinine levels less than 1,6mg/dL and ultrasonographic changes; 17 cats represented stage II, with serum creatinine levels between 1,6 and 2,8mg/dL; eight cats represented stage III/IV, with serum creatinine levels between 2,9 e 5,0mg/dL, grouped with the felines with serum creatinine levels above 5,0mg/dL. The kidneys underwent an ultrasonographic examination observing: cortical echogenicity, regularity of the contour and corticomedullary definition in the B mode; length, width and height in the longitudinal, transverse and dorsal planes in the B mode; filling of the interlobars, arcuate and interlobulars arteries by the color Doppler; and resistive index of the intrarenal vessels using the pulsed Doppler. The ratio between the length of the kidney and the luminal diameter of the aorta in normal felines and in felines with kidney disease was established. The increase in echogenicity of the cortex showed to be a relevant characteristic to be considered in the ultrasonographic evaluation of chronic kidney disease. The color Doppler showed to be an important tool in the diagnosis of the chronic kidney disease, especially when the alterations in the B mode werent expressive. The resistive index did not present itself above of the normal limits in the initial stages of the chronic kidney disease, suggesting its uselessness as a predictor of the chronic kidney disease. Changes in B mode associated with increase in resistive index may indicate a poor prognostics of the chronic kidney disease. Increase in the echogenicity of the cortical, contour irregularity, corticomedullary indefinition, reduced vascular filling detected by color Doppler and the increase in the resistive index were important elements to be considered in the diagnosis of the chronic kidney disease.

This paper details the making of Stray Cats, a University of New Orleans thesis film. It examines the process of creating the film in three parts. Part one will detail the pre-production, including the conceptualization, writing, and preparation for the project. Part two will cover the production, including the directing, production design, cinematography, and on-set operations. Part three will describe the post-production process, including the editing, color correction, sound design, and musical score. These three components will then be referenced in regards to the theme of the film, and will conclude by analyzing whether or not the final film achieves the initial vision.

Our lives would not be what they are today without animals. From the food we eat, to the clothes we wear, animals provide tangible evidence of their importance every day. But more than that, animals have shaped who we are and what we believe. Often in ways we don't see. That's what inspired me to write Being Human. This work began as an examination of how humans have altered animals to better match our imaginations and ideals, and too, the way these animals have irrecoverably altered how we live and look at the world. Consider, for example, that before they became physically useful to us in providing meat or skills or companionship, animals were central figures in our stories, mythologies, and religions. All the while, of course, these animals remained both ignorant and at the mercy of whatever we imagined--or needed--them to be. And what does all of this say about us? What can we learn about ourselves from looking at animals, and more specifically, looking at the way we treat them? In a society where animal flesh comes to us freshly packed and cleanly saran-wrapped, and pets are treated as members of our families, we tend to look at animals as one thing or another. A farm pig is not a companion animal, any more than a cat is a meal-in-waiting. At least not in our culture. We generally see what's convenient or desirable and when things get messy or complicated, we tend to look away. In so doing, we miss the opportunity to clearly see who we really are, what we're capable of, and what, if anything, we might want to change as a result. I chose four specific animals that show us different sides of ourselves. These beings are both familiar and strange, part of our everyday lives but often only found on the periphery. Each animal symbolizes one of four categories: food, pest, worker and pet. And each connects to a human need: pigs with consumption, pigeons with communication, horses with control and cats with companionship. They are arranged in this order to reflect the deepening complexity of their respective human needs--from the simplest, the need to eat, to the most complex, the need for companionship. (Arguably, control can be considered the most complex, however I chose companionship as the culminating need because it inherently involves all of the other three.) I hope if I accomplish only one thing, it is this: after reading, you see these animals--and your relationship to them--a little bit differently than before.

Addenda slip inserted in back. Includes bibliographical references (leaves 178-231). To further the understanding of the molecular mechanisms that contribute to the skeletal pathology of mucopolysaccharidosis type VI and to investigate the production of organic matrix by mucopolysaccharidosis VI osteoblasts

The initial focus of this PhD project was on comparative gene mapping. Comparative gene mapping is facilitated by consensus PCR primers which amplify homologous gene fragments in many species. As a part of an international co-ordinated programme of comparative mapping in pigs, 47 CATS (Comparative Anchor Tagged Sequence) consensus primer pairs for loci located on human chromosomes 9, 10, 20, and 22, were used for amplifying homologous loci in pigs. After optimization of PCR conditions, 23 CATS products have confirmed by comparison with homologous sequences in GenBank. A French somatic cell hybrid panel was used to physically map the 6 porcine CATS products distinguishable from rodent background product, namely ADRA1A, ADRA2A, ARSA, GNAS1, OXT and TOP1. Of these, the map location of ADRA1A and OXT showed inconsistency with the previously recognised conserved relationship between human and pig. The other four loci mapped to positions consistent with known syntenic relationships. Despite low levels of polymorphism, frequently indistinguishable rodent and porcine products in somatic hybrids and some confusion of identity of gene family members, these CATS primers have made a useful contribution to the porcine-human comparative map. The focus of the project then changed to genetic and molecular characterisation of endogenous retroviruses in pigs and their relatives. Pigs are regarded as a potentially good source of organs and tissues for transplantation into humans. However, porcine endogenous retroviruses have emerged as a possible problem as they can infect cultured human cells. Two main types of pig retrovirus, determined by envelope protein, PERV-A and PERV-B, are widely distributed in different pig breeds and a third less common type, PERV-C, has also been recognised. Endogenous retroviruses were analyzed from the Westran (Westmead transplantation) inbred line of pig, specially bred for biomedical research. Thirty-one 1.8 kb env PCR product clones were sequenced after preliminary screening with the restriction enzymes KpnI and MboI. Five recombinant clones between A and B were identified. 55% of clones (17/31) sequenced had stop codons within the envelope protein-encoding region, which would prevent the retrovirus from making full-length envelope protein recognizable by cell-surface receptors of the virus. The endogenous viruses were physically mapped in Westran pigs by FISH (Fluorescence In Situ Hybridisation) using PERV-A and PERV-B envelope clones as probes. Preliminary FISH data suggest that there are at least 22 PERVs (13 PERV-A and 9 PERV-B) and the chromosomal locations of these in the Westran strain are quite different from European Large White pigs. The sequences and mapping results of inbred Westran pig suggest that there are relatively few PERV integration sites compared with commercial pigs and further that a large proportion of clones are defective due to premature stop codons in the envelope gene. To investigate the relationship of endogenous retroviruses in peccaries and pigs, a set of degenerate primers was used to amplify peccary retroviral sequences. The sequences of two putative retroviral clones showed close homology, albeit with a 534 bp deletion, to mouse and pig retroviral sequences. Also, four non-target sequences were amplified from peccary with the degenerate retroviral primers. They are a part of the peccary cofilin gene, a SINE, and a sequence containing a microsatellite. The peccary endogenous retroviral sequences are significant in that they are the first such sequences reported in peccary species and repudiate old claims in the literature that peccaries do not have C-type retroviral sequences.

Addenda slip inserted in back. / Includes bibliographical references (leaves 178-231). / xiv, 234, [19] leaves, [56] leaves of plates : ill. (chiefly col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / To further the understanding of the molecular mechanisms that contribute to the skeletal pathology of mucopolysaccharidosis type VI and to investigate the production of organic matrix by mucopolysaccharidosis VI osteoblasts / Thesis (Ph.D.)--University of Adelaide, Dept. of Paediatrics, 2001

When modeling a neuron, modelers often focus on the values of parameters that produce a desired output. However, if these parameters are not unique, there could be a number of parameter sets that produce the same output. Thus, even though the values of the various maximum conductances, half activation voltages and so on differ, as a set they can produce the same spike height, firing rates, and so forth. To examine whether or not parameter sets are unique, a 3-compartment motoneuron model was created that has 15 target outputs and 59 parameters. Using parameter searches, over one hundred parameter sets were created for this model that produced the same output (within tolerances). Parameter values vary between parameter sets and indicate that the parameter values are not unique. In addition, some parameters are more tightly constrained than others. Principal component analysis is used to examine the dimensionality of the input and output spaces. However, neurons are more than static output generators. For example, a variety of neuromodulatory influences are known to shift parameter values to alter neuronal output. Thus the question arises as to whether this non-uniqueness extends from model outputs to the models sensitivities to its parameters. In this work, the non-unique parameter sets are further analyzed using sensitivity analyses and output correlations to show that these values vary significantly between these parameter sets. Therefore, each of these models will react to parameter variation differently. This work concludes that parameter sets are non-unique but have varying sensitivity analyses and output correlations. The ramifications of this are discussed for both modelers and neuroscientists.