Biocompatibilidade e biodegradabilidade de micropartículas de poli-lactato-co-glicolato na ceratite infecciosa em coelhos / Biocompatibility and biodegradability of poly-latic-co-glycolatic microparticles in the infectious keratitis in rabbits

Avaliaram-se a biocompatibilidade e a biodegradabilidade do sistema de liberação controlada de poli-lactato-co-glicolato (PLGA) no tratamento com ciprofloxacina das ceratites por Staphylococcus aureus em coelhos. Foram utilizados 20 coelhos, distribuídos em quatro grupos (G). Os animais dos G1, G3 e G4 foram inoculados com 2,5µL da bactéria - 108UFC, no estroma corneano. Os do G2 não receberam a aplicação do inóculo. O tratamento foi realizado com solução salina básica para os animais do G1, micropartículas de PLGA contendo ciprofloxacina nos animais dos G2 e G4 e colírio de ciprofloxacina naqueles do G3. Suabe e biópsia da superfície ocular foram coletados para cultura. Apenas um animal do G1 apresentou cultura positiva para S. aureus. Exame histológico revelou a presença bacteriana em todos os animais do G1 e em dois animais do G3. Também foi constatada reação inflamatória no local da aplicação do sistema de liberação controlada. O tratamento com micropartículas de PLGA foi eficiente no tratamento de ceratites bacterianas, ao eliminar por completo a presença do S. aureus, mas entretanto não foi completamente biocompatível e biodegradável após cinco dias.

The biocompatibility and biodegradability of the controlled delivery system of Poly-Latic-Co-Glucolatic (PLGA) in the treatment of Staphylococcus aureus keratitis with ciprofloxacin in rabbits were evaluated. Twenty rabbits divided into four groups (G) were used. G1, G3 and G4 animals were inoculated with bacterial 2.5µL (108CFU) in the corneal stroma, and G2 animals did not receive the application of inoculum. The treatment was performed with basic saline solution in G1 rabbits, micro particles of PLGA containing ciprofloxacin in G2 and G4 animals, and ciprofloxacin eye drops in G3 rabbits. Swab and biopsy of the ocular surface were collected for culture. Only one animal in G1 had positive culture for S. aureus in the processed material. Histological examination showed a bacterial presence in all animals in G1 and two animals in G3. Inflammatory reaction was noted at the application site of the controlled release. Data analysis showed that treatment with micro particles of PLGA was effective in treating bacterial keratitis, completely eliminating the presence of S. aureus, but it was not being completely biocompatible and biodegradable after five days.

Effect of image resolution manipulation in rearfoot angle measurements obtained with photogrammetry

The aim of this study was to investigate the influence of image resolution manipulation on the photogrammetric measurement of the rearfoot static angle. The study design was that of a reliability study. We evaluated 19 healthy young adults (11 females and 8 males). The photographs were taken at 1536 pixels in the greatest dimension, resized into four different resolutions (1200, 768, 600, 384 pixels) and analyzed by three equally trained examiners on a 96-pixels per inch (ppi) screen. An experienced physiotherapist marked the anatomic landmarks of rearfoot static angles on two occasions within a 1-week interval. Three different examiners had marked angles on digital pictures. The systematic error and the smallest detectable difference were calculated from the angle values between the image resolutions and times of evaluation. Different resolutions were compared by analysis of variance. Inter- and intra-examiner reliability was calculated by intra-class correlation coefficients (ICC). The rearfoot static angles obtained by the examiners in each resolution were not different (P > 0.05); however, the higher the image resolution the better the inter-examiner reliability. The intra-examiner reliability (within a 1-week interval) was considered to be unacceptable for all image resolutions (ICC range: 0.08-0.52). The whole body image of an adult with a minimum size of 768 pixels analyzed on a 96-ppi screen can provide very good inter-examiner reliability for photogrammetric measurements of rearfoot static angles (ICC range: 0.85-0.92), although the intra-examiner reliability within each resolution was not acceptable. Therefore, this method is not a proper tool for follow-up evaluations of patients within a therapeutic protocol.

Lavado broncoalveolar em eqüinos: revisão de literatura. Parte 1: Técnicas de colheita / Bronchoalveolar lavage in equine: review of the literature. part 1: collecting techniques / Lavado bronco - alveolar en el equino: revisión de la literatura parte 1: técnicas de recolección

O lavado broncoalveolar é um método que permite obter amostras de material oriundo do trato respiratório posterior. Sua principal aplicação prática na clínica médica de eqüinos é para a avaliação citológica de enfermidades inflamatórias não-infecciosas pulmonares (doença inflamatória das vias aéreas e doença pulmonar obstrutiva crônica) bem como da hemorragia pulmonar induzida pelo exercício. Embora essa técnica de diagnóstico seja bastante utilizada em outros países, ainda é pouco difundida no Brasil (o objetivo desta primeira parte da revisão é divulgar informações sobre técnicas de colheita de lavado broncoalveolar que possam ser úteis para implementação delas na rotina diagnóstica, pois é um método pouco invasivo que pode ser realizado a campo.

ABSTRACT:Bronchoalveolar lavage is a method that allows the collection of material samples from the superior respiratory tract. Its main practical application in equine medical clinic is the cytological evaluation of non-infectious infl ammatory diseases of the lungs (infl ammatory airway disease and chronic obstructive pulmonary disease), as well as exercise-induced pulmonary hemorrhage. Although this diagnosis technique is broadly used in other countries, it is not commonly applied in Brazil. The objective of the fi rst part of this review is to widespread information about collection techniques of bronchoalveolar lavage in order to aid the implementation of this diagnostic routine, for it is little invasive and may be usedin fi eld conditions

RESUMEN: El lavado bronco-alveolar es un método que permite obtener muestras de material procedente del tracto respiratorio posterior. Su principal aplicación práctica en la clínica médica de equinos es la evaluación citológica de enfermedades infl amatorias no infecciosas pulmonares (enfermedad infl amatoria de las vías aéreas y enfermedad pulmonar obstructiva crónica) así como de la hemorragia pulmonar inducida por el ejercicio. Aunque esta técnica de diagnóstico sea bastante usada en otros países, todavía está poco difundida en Brasil. El objetivo de esta primera parte de la revisión es divulgar informaciones sobre las técnicas de recolección del lavado bronco-alveolar y su posible inclusión en la rutina diagnóstica, pues se trata de un método poco agresivo y realizable en el campo.

Evaluation of alveolar macrophage function after experimental infection with equine herpesvirus-1 in horses

The role of the pulmonary alveolar macrophages (PAM) in the lung defense mechanism was evaluated in horses infected with equine hespesvirus-1 (EHV-1). Five adult horses were exposed to 10(6.6) TCID50 EHV-1 by intranasal instillation. Cytology of bronchoalveolar lavage (BAL) was performed using cytocentrifugation of samples and slides stained by Rosenfeld. Cell concentration was adjusted to 2´10(6) cells/ml, for the measurement of macrophage activity - spreading, phagocytosis of zymosan particles and release of hydrogen peroxide (H2O2). All animals were positive in virus isolation on the second, third and fifth days post-inoculation (DPI). Seroconversion was observed on the 14th DPI. Lymphocytosis was observed by BAL cytology on the 16th DPI. Measurement of macrophage activity demonstrated a marked increase in the spreading rate, on the 23rd and 30th DPI. Phagocytosis was decreased on the second DPI, and returned to levels similar to those observed before inoculation on the 23rd DPI. The amount of H2O2 released by PAM declined on day 2, but, by day 16, they returned to values similar to those observed before inoculation. The decline in PAM activity in the acute phase of disease is indirect evidence that these cells have an important role in lung defense mechanisms against this agent.

Avaliaçäo da funçäo de macrófagos alveolares em cavalos clinicamente sadios / Evaluation of alveolar macrophage function in healthy horses

Devido à importância dos macrófagos alveolares (MA) nos mecanismos de defesa pulmonar, foram realizados estudos para avaliar a atividade desses fagócitos em cavalos hígidos. Foram realizados lavados broncoalveolares (LBA) em cinco cavalos clinicamente sadios. A citologia foi realizada pela citocentrifugaçäo das ammostras e posterior confecçäo de lâminas coradas pelo método de Rosenfeld. Todas as amostras do LBA foram centrifugadas e a concentraçäo celular foi ajustada para 2x10 elevada a sexta potência células/ml, para a mensuraçäo da atividade macrofágica (testes de espraiamento, fagocitose e liberaçäo de peróxido de hidrogênio). A contagem diferencial das células presentes no LBA demonstrou a predominância de macrófagos (59,0ñ6,9 por cento). Os resultados obtidos nos testes de mensuraçäo da atividade macrofágica foram: índice de espraiamento 25,1ñ19,7 por cento, índice de fagocitose 89,4ñ6,2 por cento e liberaçäo de peróxido de hidrogênio 1,6ñ0,3nmoles/2x10 elevado a quinta potência células (sem PMA - phorbol 12-myristate 13-acetate) e 1,8ñ0,4nmoles/2x10 elevado a quinta potência células (com PMA). Os resultados demonstraram um padräo de atividade para MA de cavalos hígidos, os quais apresentaram índices de ativaçäo mesmo sem elicitaçäo prévia, indicando que as técnicas utilizadas foram adequadas para tal propósito

Avaliação citológica de lavados traqueobrônquico e broncoalveolar em cavalos clinicamente sadios pelo método de coloração de Rosenfeld

Comparisons of tracheobronchial (TBW) and bronchoalveolar wash (BAL) cytologies, using respectively 25 and 5 normal horses were made. Differential cell counts were made on both TBW and BAL fluids on Rosenfeld-stained cytocentrifuged preparations. Macrophages were the predominant cell types seen in both fluid samples and these findings suggest the possibility of an adequate sampling of the distal air passages. Differences were found in lymphocytes and macrophages between TBW and BAL cytologies. The main cell types in BAL fluids were macrophages (54.78±15.92%) and lymphocytes (33.70±14.41%). On the other hand, lymphocytes and macrophages were found in 2.91±3.44% and 81.51±11.06% of cell types in TBW fluids, respectively. Also, there were absences of epithelial and goblet cells in BAL cytology, and these explain the adequacy of the lower respiratory tract sampling, performed without causing trauma of proximal air passages. Other cell types were found in few numbers and no differences were found between TBW and BAL fluids. Differences were not detected concerning morphological characteristics between TBW and BAL preparations.

Foram realizados estudos comparativos dos exames citológicos obtidos a partir de amostras dos lavados traqueobrônquico (LTB) e broncoalveolar (LBA) de cavalos clinicamente sadios. Foram colhidas 25 amostras pela técnica do LTB (25 animais) e 25 amostras pela técnica do LBA (5 animais). A suspensão celular dos lavados foi centrifugada, confeccionando-se lâminas coradas pelo método de Rosenfeld. A contagem diferencial mostrou predominância de macrófagos em ambas as técnicas de colheita, confirmando a possibilidade de sua utilização para obter amostras celulares das vias aéreas distais. Observou-se diferença na proporção de macrófagos e linfócitos entre as técnicas dos lavados traqueobrônquico e broncoalveolar. As porcentagens de macrófagos e linfócitos observadas foram de 81,51±11,06% e 2,91±3,44% no LTB e de 54,78±15,92% e 33,70±14,41% no LBA, respectivamente. Também notou-se ausência de células epiteliais cilíndricas e produtoras de muco em LBA, indicando que essas amostras são provenientes da população alveolar e que esta técnica não traumatizou as vias aéreas proximais. Os demais tipos celulares foram identificados em proporções semelhantes e em pequenas quantidades em ambas as técnicas. As características morfológicas dos tipos celulares foram semelhantes nas duas técnicas.