RESUMO
To identify the protein encoded by the L7 region of bovine adenovirus-3 (BAdV-3), specific antisera were raised by immunizing rabbits with bacterial fusion proteins encoding the N-terminus or C-terminus of the BAdV-3 fiber protein. Immunoprecipitation and Western blot analysis confirmed that the fiber is expressed as a 102 kDa glycoprotein, which is localized to the nucleus of infected cells. To identify the nuclear localization signals (NLS), BAdV-3 fiber deletion mutants and GFP/beta-galactosidase fusion proteins were expressed in transfected cells, and subcellular localization was visualized by immunofluorescence microscopy. Analysis of deletion mutants localized the NLS to the N-terminal 41 amino acids. Analysis of the N-terminal 41 amino acids identified a cluster of basic residues between amino acid 14 and 20. Substitution of the basic residues (16KAKR19) with acidic residues (16EAEE19) resulted in the accumulation of fiber in the cytoplasm. However, 16KAKR19 or 12VYPYKAKRPNI22 were not sufficient for efficient transport of a cytoplasmic protein GFP/beta-galactosidase to the nucleus. The recombinant BAdV-3 expressing mutant fiber containing 16EAEE19 instead of 16KAKR19 was unable to replicate efficiently in Madin-Darby bovine kidney cells, suggesting that the NLS of fiber carries out important in vivo functions.
Assuntos
Proteínas do Capsídeo/genética , Proteínas do Capsídeo/metabolismo , Núcleo Celular/metabolismo , Mastadenovirus/metabolismo , Substituição de Aminoácidos , Animais , Western Blotting , Bovinos , Células Cultivadas , Chlorocebus aethiops , Citoplasma/química , Genes Reporter , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Imunoprecipitação , Mastadenovirus/genética , Microscopia de Fluorescência , Sinais de Localização Nuclear/genética , Proteínas Recombinantes de Fusão/metabolismo , Deleção de Sequência , Replicação Viral , beta-Galactosidase/genética , beta-Galactosidase/metabolismoRESUMO
50 cases of isolated meniscal injuries of the knee were evaluated and managed arthroscopically. 56% of the cases were in 25-35 year age group. In 80% of the cases military training and contact sports was the mode of injury. Maximum (42%) patients reported late (> 2 years) after the injury. On presentation, they had an average Lysholm knee score of 53.76. Medial meniscal lesion was seen in 74%. Commonest pattern of tear encountered was a longitudinal tear (40%). Depending on pattern and extent of lesion, partial meniscectomy (60%), subtotal meniscectomy (14%) and total meniscectomy (26%) were performed. Patients were followed up at six months and one year. The average Lysholm score at 6 months was 83.3 and at one year 79.5. At one year, the patient satisfaction level was 82% and patients who underwent partial meniscectomy had the best results.
