Microbiological profile of donor corneas stored for tectonic transplantation purposes in rabbits / Microbiologia de amostras de bancos de córneas destinadas a transplantes, em coelhos

This study aimed to evaluate the microbiota of donor rabbit corneas stored for tectonic transplantation purposes. Swabs from both corneas of 20 rabbits were carefully collected and submitted to microorganism isolation and identification. After this first swab collection, rabbits were euthanized for reasons other than this project and the eyes were enucleated. The corneas were collected and stored to compose the cornea tissue bank. Corneas were stored in a 0.3% tobramycin solution at -20ºC. After 30 days, the corneas were thawed at room temperature and removed from the antibiotic. New swabs were obtained from the corneas and submitted to microorganism isolation and identification. Gram positive organisms were predominant in the rabbit corneal flora before storage and the Staphylococcus sp. was the most common microorganism isolated from those samples. No growth was observed on the samples collected after storage. The methods used for collection and storage of the corneas were efficient to constitute a sterile donor corneal tissue bank.

Analisaram-se córneas armazenadas para transplantes tectônicos usando-se suabes coletados de 20 coelhos, visando ao isolamento e à identificação de microrganismos. Após a coleta das amostras, os coelhos foram submetidos à eutanásia, por razões alheias ao estudo, e enucleados. As córneas foram coletadas e armazenadas a fim de se constituir o banco de córneas. O armazenamento deu-se em solução de tobramicina 0,3% a -20ºC, por 30 dias. Após esse período, as córneas foram descongeladas à temperatura ambiente e removidas da solução de antibiótico. Novos suabes foram coletados e submetidos ao isolamento e à identificação dos microrganismos. A flora corneal mostrou-se predominantemente composta por bactérias Gram positivas, sendo o Staphylococcus sp. o mais identificado. Não se verificou crescimento de colônias bacterianas ou fúngicas nas amostras após o armazenamento. Considerando-se a maneira como a pesquisa foi concebida e as injunções do meio em que ela foi realizada, há como admitir, pela ausência de crescimento microbiano nas amostras armazenadas, que a técnica de armazenamento empregada é segura para a estocagem de córneas destinadas a transplantes.

Ultrasonographic analysis of senile cataractous lens of dogs and its correlation to phacoemulsification / Análise ultrassonográfica da lente cataratogênica senil de cães e sua correlação com parâmetros de facoemulsificação

This study aimed to evaluate the effectiveness of a computer-assisted ultrasound analysis of senile dogs cataractous lens and identify its correlation to phacoemulsification surgical parameters. Simultaneous A- and B- mode ultrasonic evaluation of the lens was performed on 15 dogs with cataract. A computer-assisted image analysis was used to objectively calculate the lens echogenicity. Following ultrasonographic examination the cataracts were removed by phacoemulsification and the ultrasonographic and phacoemulsification findings were compared to identify possible correlations. Lens echogenicity obtained by the computer-assisted ultrasonographic analysis correlated to the phacoemulsification time, such that the whiter the ultrasound image, the longer was the phacoemulsification time. The B-mode ultrasonography was helpful for the prediction of lens opacity location, but not predictive of lens hardness.

Avaliou-se a eficácia da análise ultrassonográfica, assistida por computador, da lente cataratogênica senil de cães e identificou-se sua correlação com parâmetros da facoemulsificação. A avaliação ultrassonográfica da lente em modos A e B simultâneos foi realizada em 15 cães portadores de catarata. Procedeu-se à análise das imagens, assistida por computador. Posteriormente, os olhos selecionados foram submetidos à facoemulsificação. Os achados ultrassonográficos e a facoemulsificação foram comparados visando à identificação de possíveis correlações. A ecogenicidade obtida por meio da análise ultrassonográfica correlacionou-se diretamente com o tempo de facoemulsificação, de maneira que quanto mais branca a imagem ultrassonográfica, maior o tempo de facoemulsificação. A ultrassonografia em modo-B auxiliou na predição da localização das opacidades lenticulares, porém não auxiliou na predição da dureza da lente.

Newborn screening for lysosomal storage disorders.

Lysosomal storage disorders (LSD) represent a group of over 40 distinct genetic diseases with a total incidence of approximately 1:7,000 births. Bone marrow transplantation and enzyme replacement therapy are currently in use for the treatment of some disorders and new forms of enzyme and gene replacement therapy are actively being researched. The effectiveness of these therapies, particularly for the LSD involving the central nervous system and bone pathology, will rely heavily upon the early diagnosis and treatment of the disorder, before the onset of irreversible pathology. In the absence of a family history the only practical way to detect these disorders will be by a newborn screening program. One common feature of these disorders is an increase in the number and size of lysosomes within the cell from approximately 1% to as much as 50% of total cellular volume. Associated with this, is a corresponding increase in some lysosomal proteins. We propose that the measurement of one or more of these proteins in blood spots taken from Guthrie cards, will form the basis of a newborn screening program, for the detection of all LSD. We have identified a number of lysosomal proteins as potential markers for LSD. The level of these proteins has been determined in blood spots taken from Guthrie cards and in plasma samples from over 300 LSD affected individuals representing 25 disorders. Based on these results we have proposed a strategy for a newborn screening program involving a two tier system, utilizing time resolved fluorescence immunoquantification of the protein markers in the first tier, followed by tandem mass spectrometry for the determination of stored substrates in the second tier assays.