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1.
We studied and characterized the collagenolytic matrix metalloproteinases (MMP-8 and MMP-13) in the pathogenesis of canine pulmonary eosinophilia (PE). Twenty dogs with PE and 16 healthy control dogs underwent similar clinical examination and collection of bronchoalveolar lavage fluid (BALF). Analyses of total cell and differential cell counts and collagen I degradation with and without aminophenyl mercuric acetate (APMA) treatment were performed. Correlations between cell counts and percentage of degraded collagen I in BALF were studied. Collagenase activity detected in BALF was characterized by Western immunoblotting for collagenase-2 (MMP-8) and collagenase-3 (MMP-13), and their cellular location was studied by immunocytochemical means. Collagenolytic activity was significantly increased in cell-free and native BALF of PE dogs compared to healthy controls. APMA treatment had no significant effect on BALF collagenase activity, indicating that collagenolytic activity occurred in diseased BALF in vivo in active form. Western immunoblotting identified the presence of MMP-8 and MMP-13 immunoreactivities, of which the latter was converted to active form. Major immunoreactivity for MMP-8 was observed in macrophages and epithelial cells, and major immunoreactivity for MMP-13 was observed in macrophages. A significant positive correlation was noted between the percentage of degraded collagen I and the counts of eosinophils, macrophages, lymphocytes, and mast cells. These findings suggest that the up-regulation of collagenolysis eventually contributes to pulmonary tissue destruction in canine PE.  相似文献   

2.
OBJECTIVE: To determine reference values for cytologic examination results of bronchoalveolar lavage fluid (BALF) and to investigate effects of repeated lavages on pulmonary health and on results of cytologic examination of BALF in dogs. ANIMALS: 16 healthy adult Beagles. PROCEDURE: All dogs underwent pulmonary lavage to obtain BALF. Eleven dogs were repeatedly lavaged 6 times at 5- to 7-week intervals. Analyses for total and differential cell counts and for viability of cells before and after cell processing were performed. Arterial blood gas analysis before and after bronchoalveolar lavage was used to study the safety of the lavage procedure. Histologic and radiologic examinations were used to study effects of repeated lavages on pulmonary health. RESULTS: Mean (+/- SD) cell count was 104 +/- 69 cells/microl, comprising 75 +/- 7% alveolar macrophages, 13 +/- 6% lymphocytes, 5 +/- 4% neutrophils, 4 +/- 5% eosinophils, 2 +/- 2% mast cells, 0.6 +/- 0.7% epithelial cells, and 0.3 +/- 0.4% plasma cells. Centrifugation of samples and washing of cells caused significant cell loss (59 +/- 13%). Repeated lavages did not cause significant variations in cell counts of BALF or results of arterial blood gas analysis, thoracic radiography, or histologic examination of pulmonary specimens. Only a moderate, although significant, decrease in arterial oxygen content was observed after bronchoalveolar lavage. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis indicated that several lavages performed at 5- to 7-week intervals can safely and reliably be used to study the kinetics of pathologic processes in pulmonary tissues or for evaluation of therapeutic efficacy.  相似文献   

3.
Background: Idiopathic pulmonary fibrosis (IPF) is a chronic, interstitial lung disease primarily affecting West Highland White Terriers (WHWTs). Objective: To describe the clinicopathological and diagnostic imaging features in WHWTs with IPF. Animals: Twelve WHWTs with IPF and 14 healthy control WHWTs. Method: Prospective study. Clinical signs and findings of physical examination, blood and arterial blood gas analyses, radiography, high‐resolution computed tomography (HRCT), bronchoscopy and bronchoalveolar lavage (BAL) of IPF dogs were obtained and compared with controls. Histopathologic changes in IPF dogs were evaluated. Results: Mean partial pressure of oxygen was significantly lower in IPF (mean ± SD, 65.5 ± 15.4 mmHg) than in controls (99.1 ± 7.8 mmHg, P<.001). The alveolar‐arterial oxygen gradient was significantly higher in IPF (50.1 ± 17.3 mmHg) than in controls (17.5 ± 4.9 mmHg, P<.001). In HRCT, ground glass opacity (GGO) was detected in all IPF dogs, traction bronchiectasis in 4, and honeycombing in 1. Bronchoscopic airway changes were noted in all IPF dogs. On BAL fluid (BALF) cytology, the total cell count (TCC) was higher in IPF dogs, and the numbers but not the percentages of macrophages, neutrophils, and mast cells were increased. On histopathology, multifocal or diffuse interstitial fibrosis, type II pneumocyte hyperplasia, prominent intraalveolar macrophages, distortion of alveolar architecture, and emphysematous change were detected. Conclusion and Clinical Importance: IPF causes substantial hypoxemia. In HRCT, GGO is a consistent finding. IPF dogs have concurrent airway changes and an increase in BALF TCC.  相似文献   

4.
Inflammation causes epithelial cell sloughing and basement membrane (BM) exposure in canine pulmonary eosinophilia (PE), leading to degradation of the epithelial cell attachment component, laminin-5 gamma2-chain, into small molecular weight fragments. The subsidence of inflammation after treatment down-regulates degradation. Laminin-5 gamma2-chain levels and molecular forms in bronchoalveolar lavage fluid (BALF) were analysed semiquantitatively by Western immunoblotting to compare PE affected (n=20) and healthy dogs (n=16) as well as PE dogs (n=6) before and after corticosteroid treatment. PE dogs expressed significantly elevated levels of total (P<0.01), 36 kDa (P<0.05) and 53 kDa (P<0.05) laminin-5 gamma2-fragments. The 36 Da fragment decreased significantly (P<0.05) after treatment. The laminin-5 gamma2-chain degradation products may be linked to epithelial cell sloughing and BM exposure or healing.  相似文献   

5.
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6.
OBJECTIVE: To determine whether bronchial brushings from dogs with chronic cough have increased numbers of goblet cells and WBCs, compared with numbers for healthy dogs, or have differing WBC populations, compared with populations in bronchoalveolar lavage (BAL) fluid obtained from dogs with chronic cough. ANIMALS: 9 healthy dogs and 10 dogs with chronic cough. PROCEDURE: Specimens were collected by use of bronchoscopy. Cellular composition was determined for brushings, and results from dogs with chronic cough were compared with those from healthy dogs. Cellular composition of brushings was compared with composition of BAL obtained from dogs with chronic cough. RESULTS: Brushings from healthy dogs contained a median of 2.9 x 10(6) epithelial cells, comprising 100% epithelial cells (96% ciliated, 3% goblet, and 1% other) and no WBCs. Brushings from dogs with chronic cough had 4.5 x 10(6) epithelial cells, comprising 93% epithelial cells (86% ciliated, 2% goblet, and 12% other). Dogs with chronic cough had significantly greater percentages of WBCs (7%) and neutrophils (6%), compared with values for healthy dogs. Five dogs with chronic cough had no neutrophilic inflammation evident in BAL, but 4 of these had evidence of neutrophilic inflammation in brushings. CONCLUSIONS AND CLINICAL RELEVANCE: Neutrophils, but not goblet cells, were increased in brushings from dogs with chronic cough. Analysis of bronchial brushings provides information about airway inflammation that differs from that found by examination of BAL in some dogs with chronic cough and is a more sensitive indicator of airway inflammation than cytologic examination of BAL in these dogs.  相似文献   

7.
Gelatinolytic activity was analysed to study whether elevated activity previously found at the tracheal level of the respiratory tract of horses with chronic obstructive pulmonary disease (COPD) could also be found at the lower part of the respiratory tract. Furthermore, presence and significance of the gelatinolytic matrix metalloproteinases (MMPs) MMP-2 and MMP-9 in respiratory secretions of healthy and COPD horses were determined. Elevated gelatinolytic matrix metalloproteinases were detected in bronchoalveolar and tracheobronchial secretions from COPD horses. The main pathologically elevated MMP was characterised to be MMP-9. Significantly increased MMP-9 activities as measured by gelatin zymography and Western blotting were found in all the respiratory samples from COPD horses compared to healthy horses. Elevation of active MMP-9 paralleled with increased gelatinase-associated lipocalin levels. Bronchoalveolar lavage fluid (BALF) epithelial cells, macrophages, neutrophils and lymphocytes expressed MMP-9 immunoreactivity demonstrated by immunocytochemistry and MMP-9 mRNA was expressed by bronchial epithelial cells of lung tissue section shown by in situ hybridisation. MMP-2 seems not to play a major role in chronic lung inflammation. No clear differences in MMP-2 or MMP-14 (a potent MMP-2 activator) levels were found when comparing the samples from COPD or healthy horses. These results suggests that MMP-9 could serve as a potential diagnostic marker for the active ongoing tissue remodelling in the acute phase of equine COPD. Increased gelatinolytic activity could be found at both tested respiratory tract levels. Therefore, tracheal epithelial lining fluid (TELF) samples can usefully serve as diagnostic material for detection of increased levels of the main gelatinolytic MMP, MMP-9, representing the entire diseased lung.  相似文献   

8.
Bronchoalveolar lavage (BAL) fluid was analyzed in healthy horses, using different lavage fluid volumes and lung sites. The only significant difference in the cellular composition of BAL fluid between the right and left lungs was the mast cell numbers, which were significantly higher in the left lung. Total cell count ranged from 34 to 330 cells/microliter for the right lung and 43 to 330 cells/microliter for the left lung. Percentage of neutrophils ranged from 1 to 7% in the right lung and 1 to 5% in the left lung. The small-volume (50 ml) lavage had a greater percentage of neutrophils and a lesser percentage of mast cells in the large-volume (350 ml) lavage. Statistical difference in the composition of BAL fluid recovered was not detected between the 3 sequential 100-ml lavages and a single 300-ml lavage, except that macrophages were significantly higher in the 3 sequential 100-ml lavages. Values for BAL fluid analysis in healthy horses have varied considerably and this variation is from a failure to adhere to any standard technique for volume of fluid infused.  相似文献   

9.
Reference values of cellular and non-cellular components in the bronchoalveolar lavage fluid (BALF) were established from the BALF specimens obtained from 52 healthy pigs. Using urea as an endogenous marker of dilution, the reference values in the epithelial lining fluid (ELF) were calculated: total cell count 2.71 x 10(9) - 56.49 x 10(9) litre(-1) ELF, alveolar macrophages 2.02 x 10(9) - 49.91 x 10(9) litre(-1) ELF, lymphocytes 0.10 x 10(9) - 4.74 x 10(9) litre(-1) ELF, polymorphonuclear neutrophils 0.01 x 10(9) - 3.48 x 10(9) litre(-1) ELF, protein 0.10 - 13.13 g litre(-1) ELF, lactate dehydrogenase 127-1843 Units litre(-1) ELF, and alkaline phosphatase 86-994 Units litre(-1) ELF. The problems of quantification of BALF components are discussed and a standardized lavage protocol in swine is described, which is essential for the interpretation of diagnostic findings and for the comparison of different BALF specimens.  相似文献   

10.
Background: Bronchoalveolar lavage (BAL) allows cell recovery from the lower respiratory tract; differential cell counts of BAL fluid gives important information in the assessment of various bronchial and pulmonary diseases. To the best of our knowledge no study has investigated the relation between the number of cells counted and the reproducibility of BAL fluid differential cell counts. Objective: The purpose of this study was to investigate using statistical methods how many cells should be counted in cytocentrifuged BAL fluid preparations in order to obtain a reliable enumeration of each cell type. Methods: BAL fluid samples from dogs with suspected bronchopulmonary disease were obtained during fiberoptic bronchoscopy with a standardized protocol. Differential cell counts were performed on May–Grünwald–Giemsa‐stained cytocentrifuged preparations by 2 independent observers. Reproducibility for the enumeration of each cell type was expressed as the intraclass correlation coefficient. We considered a threshold level of ≥0.90 to be high and a threshold level of ≥0.85 to be adequate. Results: Forty BAL fluid samples were included in the study. For neutrophils, alveolar macrophages, and eosinophils high reproducibility was reached by counting 200 cells; adequate reproducibility was reached for lymphocytes and bronchial epithelial cells by counting 500 cells. Conclusions: A 500‐cell differential count is required for all types of cells to be quantified with adequate reproducibility in canine cytocentrifuged BAL fluid samples.  相似文献   

11.
Bronchoalveolar lavage (BAL) is a procedure that retrieves cells and other elements from the lungs for evaluation, which helps in the diagnosis of many pulmonary diseases. The aims of this work were to perform this procedure in dogs in the acute and chronic phases of an Angiostrongylus vasorum infection for cytological analysis and to evaluate the potential of this technique as a diagnostic method for this lung-heart worm. The BAL procedure was performed through the use of an endotracheal tube on seven A. vasorum infected dogs and on five non-infected dogs lined as a control group. Sixty days post-infection (dpi) active and live larvae were retrieved from the bronchoalveolar fluid (BALF) of all infected dogs. Furthermore, in one animal it was possible to retrieve larvae in its BALF before the pre-patent period. This work reports that the A. vasorum infection resulted in an increase of relative neutrophils and eosinophils counts. In contrast, there was a significant decrease in the alveolar macrophage relative count in infected animals from 60 to 330 dpi. This study shows that the BAL is an accurate technique for the diagnosis of canine angiostrongylosis. Moreover, the technique allows us to retrieve cells and other elements that line the lung surface for cytological evaluation, which provides information about inflammatory diseases, and the diagnosis and prognosis of pulmonary parasites such as A. vasorum.  相似文献   

12.
Differences in the cytological interpretation of bronchoalveolar lavage fluid (BALF) after cytospin preparation (CP) or manual smearing of pelleted cells preparation (MSP) were investigated in client-owned dogs and cats with inflammatory or infectious lower respiratory disease. Bronchoalveolar lavage fluid from healthy cats was also examined. With MSP, cell lysis was more frequently observed, and cellular distribution was more heterogeneous throughout the slide. When samples from healthy and diseased animals were considered together, a significantly greater percentage of neutrophils was seen on CP than on MSP slides (P<0.002). Cytospin preparations were considered of better quality in all individual comparisons. Cytospin preparation is advised in the evaluation of BALF with low total cell count. When only MSPs are evaluated, clinicians should be aware that differential neutrophil counts may underestimate the counts found on CP slides.  相似文献   

13.
In diagnosing inflammatory airway disease (IAD) in performance horses, a histamine bronchoprovocation (HBP) test is often performed. In previously published studies, HBP is usually undertaken prior to cytological examination of the bronchoalveolar lavage (BAL) cells. The purpose of this study was to determine if HBP alters (1) the total nucleated cell numbers and distribution in BAL fluid (BALF) and (2) the mRNA and protein concentrations of selected cytokines in BAL cells and BALF, respectively. BALF was initially collected endoscopically from the right middle or diaphragmatic lung lobe in eight healthy young Standardbred horses. Five to six days later, HBP was performed by aerosolization of histamine (8mg) over a 2min period. BALF was again collected within 2-4h of the HBP from the left middle or diaphragmatic lung lobe. In both samples, total and differential WBC counts were obtained. The gene expressions of interleukin-4 (IL-4), IL-8, interferon-gamma (IFN-gamma) and beta-actin in BAL cells were measured using real-time RT-PCR. The cytokine protein concentrations were measured in the BALF using ELISA. HBP was not associated with either a change in the total BAL cell number or in the distribution of the BAL cells. BAL cell expression of IL-4, IL-8 and IFN-gamma, detected in all samples with the exception of IL-4 in one horse (post-HBP), was not altered as a result of HBP. HBP was not associated with a significant change in IL-8 or IFN-gamma concentrations in the BALF. IL-4 protein was undetectable in BALF either prior to or following HBP. We conclude that HBP can precede BALF collection performed within 2-4h of the former without affecting selected parameters analysed in the BAL cells or BALF.  相似文献   

14.
The effect of strenuous exercise on the functional capacity of pulmonary alveolar macrophages (PAM) and bronchoalveolar lavage-derived lymphocytes was determined in eight horses prior to and after 7 weeks of training. Strenuous exercise had no effect on the total cell count or the percentage of live cells in bronchoalveolar lavage (BAL) samples prior to or following training. However, training was associated with a significant increase in the total cell count of pre-exercise BAL samples and a significant reduction in the percentage of live cells in post-exercise samples. Strenuous exercise was associated with impaired phagocytosis by PAM after 7 weeks of training but had no effect on similar samples obtained from untrained horses. The oxidative burst activity of PAM was significantly increased following strenuous exercise for both untrained and trained horses. BAL -derived lymphocyte oxidative burst was similarly affected following training. These results suggest that strenuous exercise and training may influence pulmonary immune cell function.  相似文献   

15.
Mast cells normally are present in equine bronchoalveolar lavage fluid (BALF), but usually represent <2% of all cells in healthy horses. An increased percentage of mast cells has been associated with airway hyperactivity and inflammatory airway diseases, but marked differences are reported between studies in normal and diseased horses. Because an abnormal mast cell count may be of clinical relevance, we compared the ability of a fast Romanowsky method to stain mast cell granules with that of 3 metachromatic stains: automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains. The BALF cells from 24 horses were studied. A differential cell count was performed blindly on 400 cells. The percentages of mast cells obtained were analyzed by means of repeated-measures analysis of variance and Fischer's PLSD test. The Bland and Altman method was used to assess agreement among stains. The mean percentage of mast cells in BALF was significantly lower with the fast Romanowsky than with the automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains. With the fast Romanowsky stain, the metachromatic granules of mast cells were not stained, and their identification was based on morphologic criteria. Toluidine blue staining allowed detection of the highest mean percentage of mast cells, but was inadequate for performing a differential cell count on other cell types. In conclusion, fast Romanosky stain may be inadequate for detection of mast cells in equine BALF, whereas automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains provide metachromatic staining of mast cell granules.  相似文献   

16.
The objective of this study was to investigate the normal cytological evaluation of bronchoalveolar lavage (BAL) fluid in healthy adult ferrets (N = 12). These ferrets underwent bronchoscopy and BAL using sterile saline [1.5 mL/kg body weight (BW)]. Percentage of fluid recovered, total leukocyte count, differential leukocyte count, and cell count of the epithelial lining fluid (ELF) were determined. The mean percentage of lavage volume recovered from the right lung and left lung were 67.8 ± 14.9% and 69.7 ± 20.0%, respectively. Gender (P = 0.12) and weight (P = 0.17) did not significantly affect the mean percentage of recovered volume. The mean percentage of recovered volume (P = 0.47) and the mean leukocyte count (P = 0.17) from the right and left lung were not significantly different. Macrophages were the main leukocyte component of the lavages, followed by neutrophils, lymphocytes, and eosinophils. The mean proportion of ELF in BAL fluid was 9.3 ± 3.7% v/v. Bronchoscopy is clinically useful for collecting good quality BAL samples for cytological analysis in ferrets. The leucocyte differential was established, which may help veterinarians to make better clinical decisions when treating respiratory disease. Further studies are required with a larger group in order to establish the healthy reference intervals for BAL values in ferrets.  相似文献   

17.
The effect of systemic administration of glucocorticoids was evaluated on 3 populations of macrophages obtained from healthy dogs. Phagocytic and Fc-receptor activities were determined for mononuclear phagocytes from blood and pulmonary and peritoneal lavage fluids. Samples were collected from 12 dogs before treatment and again on the same dogs after glucocorticoid administration. Thirty or more days were allowed between treatment periods. Twelve hours after combined prednisolone sodium succinate and dexamethasone sodium phosphate administration, the percentage of phagocytosing cells decreased for blood monocytes and increased for pulmonary macrophages. The percentage of pulmonary macrophages positive for erythrocyte antibody-rosette formation (Fc-receptor activity) increased. After 7 days of oral administration of prednisone, the percentage of phagocytosing peritoneal macrophages increased, whereas the percentage of blood monocytes with Fc-receptor activity increased. Results indicate that significant (P less than 0.05) changes in macrophage function occur after systemic administration of the glucocorticoid doses used in this study. Also, the effect of systemic administration of glucocorticoids on mononuclear phagocytes varies, depending on the specific cell location.  相似文献   

18.
Lai SC  Chen KM 《Veterinary parasitology》2007,150(1-2):122-127
The relationship between proteolytic enzymes and hematological response to infection was studied in five 1-month-old dogs inoculated experimentally with 2000 eggs of Toxocara canis. Moderate leukocytosis and marked eosinophilia occurred 14 days post-infection with T. canis. Plasminogen activators (PAs) and matrix metalloproteinase-9 (MMP-9) activity in serum was significantly different in dogs infected with T. canis, compared with controls. Urokinase-type PA (uPA) activity was positively correlated with eosinophilia, and tissue-type PA (tPA) and MMP-9 activity was negatively correlated with eosinophilia. However, there was no correlation between inflammation and MMP-2. The use of uPA, tPA or MMP-9 proteolytic enzymes as laboratory reference markers for toxocarosis requires further study.  相似文献   

19.
BACKGROUND: Various diagnostic tests have been used to assign a clinical stage to dogs with lymphoma. As more sensitive staging methods are introduced, dogs are reclassified as having a higher disease stage, thereby affecting comparisons of dogs across differently staged clinical trials, and possibly, prognosis. HYPOTHESIS: The addition of more sensitive staging tests causes stage migration in dogs with lymphoma. ANIMALS: Fifty-nine client-owned dogs with previously untreated cytologically or histologically confirmed lymphoma METHODS: For every dog, the World Health Organization stage classification (I-V) was based on 5 groupings of various diagnostic tests: A (physical examination [PE] and quantitative blood count [QBC]), B (PE, QBC, thoracic and abdominal radiographs), C (PE, complete blood count with blood-smear evaluation [CBC], thoracic and abdominal radiographs), D (PE, CBC, thoracic radiographs, abdominal ultrasound), and E (PE, CBC, thoracic radiographs, abdominal ultrasound, and bone-marrow cytology). Dogs were treated with doxorubicin-based protocols. RESULTS: There was migration between all of the staging methods except D to E. However, the stage was not a predictor of remission rate, remission duration, or survival, regardless of staging method used. CONCLUSIONS AND CLINICAL IMPORTANCE: These data emphasized the need for standardized methods to determine the clinical stage in dogs with lymphoma.  相似文献   

20.
Background: Cerebrospinal fluid (CSF) pleocytosis recently was associated with the severity of neurologic signs in dogs with intervertebral disc disease (IVDD). Hypothesis/Objectives: To look for an association among CSF cell counts, total protein concentration, and severity of neurologic signs at presentation with outcome in dogs with acute thoracolumbar IVDD. Our hypothesis was that CSF total nucleated cell count (TNCC) and percentage cell types would be associated with the severity of spinal cord damage and therefore with both the presenting clinical signs and the prognosis of affected dogs. Animals: Fifty‐four dogs with acute nonambulatory thoracolumbar IVDD were evaluated. Methods: Retrospective study. Signalment, neurologic grade, CSF TNCC, protein concentration, red blood cells count and differential cell percentages, and short‐ and long‐term outcomes were evaluated. Results: CSF pleocytosis (>5 cells/μL) was present in 54% of dogs and was positively associated with neurologic grade at presentation and with postoperative time to regaining ambulation. Neutrophils were observed most frequently. The percentage of CSF macrophages and macrophage to monocyte ratio were higher (P= .001, for both) in dogs presented without deep pain sensation (DPS) that did not regain ambulation. Receiver operator characteristics curve analysis yielded a cut‐off point of 13% macrophages with a sensitivity and specificity of 100 and 83%, respectively, for prediction of a negative outcome. Conclusions and Clinical Importance: CSF pleocytosis is positively associated with the severity of spinal cord damage in dogs with thoracolumbar IVDD. The percentage of CSF macrophages can be used as a prognostic indicator for regaining ambulation in dogs that have lost DPS.  相似文献   

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