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BACKGROUND: Ex vivo lung perfusion (EVLP) constitutes a tool with great research potential due to its advantages over in vivo and in vitro models. Despite its important contribution to lung reconditioning, this technique has the disadvantage of incurring high costs and can induce pulmonary endothelial injury through perfusion and ventilation. The pulmonary endothelium is made up of endothelial glycocalyx (EG), a coating of proteoglycans (PG) on the luminal surface. PGs are glycoproteins linked to terminal sialic acids (Sia) that can affect homeostasis with responses leading to edema formation. This study evaluated the effect of two ex vivo perfusion solutions on lung function and endothelial injury. METHODS: We divided ten landrace swine into two groups and subjected them to EVLP for 120 min: Group I (n = 5) was perfused with Steen® solution, and Group II (n = 5) was perfused with low-potassium dextran-albumin solution. Ventilatory mechanics, histology, gravimetry, and sialic acid concentrations were evaluated. RESULTS: Both groups showed changes in pulmonary vascular resistance and ventilatory mechanics (p < 0.05, Student's t-test). In addition, the lung injury severity score was better in Group I than in Group II (p < 0.05, Mann-Whitney U); and both groups exhibited a significant increase in Sia concentrations in the perfusate (p < 0.05 t-Student) and Sia immunohistochemical expression. CONCLUSIONS: Sia, as a product of EG disruption during EVLP, was found in all samples obtained in the system; however, the changes in its concentration showed no apparent correlation with lung function.
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Lesão Pulmonar , Ácido N-Acetilneuramínico , Animais , Suínos , Respiração , Perfusão , Pulmão , Modelos TeóricosRESUMO
Ninety crossbreed bulls (349.5 ± 8.25 kg initial weight) were used in an 87day trial to compare the effects of a blend of essential oils plus 25-hydroxy-Vit-D3 (EO + HyD) versus the combination of monensin with virginiamycin (MON + VM) on feedlot growth performance and carcass characteristics. Dietary treatments (nine replicates/treatment) were supplemented with 40 mg/kg diet dry matter of MON + VM (equal parts) or with 120.12 mg/kg diet dry matter of a combination of standardized mixture of essential oils (120 mg) plus 0.12 mg of 25-hydroxy-vitamin-D3 (EO + HyD). There were no treatment effects on dry matter intake (DMI, p = 0.63). However, the coefficient of variation in day-to-day DMI was greater for EO + HyD than for MON + VM (11.4% vs. 3.88%, p = 0.04). There were no treatment effects (p ≥ 0.17) on daily weight gain, gain-to-feed ratio, and estimated dietary net energy. Cattle supplemented with EO + HyD had greater Longissimus muscle area (7.9%, p < 0.01) and estimated retail yield (1.6%, p = 0.03), and tended to have heavier (1.7%, p = 0.10) carcass weight. Differences among treatments in dressing percentage, fat thickness, kidney−pelvic−heart fat, and marbling score were not appreciable (p > 0.10). It is concluded that growth performance response and dietary energetic are similar for finishing cattle supplemented with EO + HyD vs. MON + VM. However, compared with MON + VM, supplementation with EO + HyD during the finishing phase may improve carcass Longissimus area and carcass yield.
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BACKGROUND: Tracheal stenosis (TS) is a complication of prolonged intubation, tracheotomy, and tracheal surgery that compromises the vascular supply. Animal models are essential for studying its pathophysiology and the effect of interventions. OBJECTIVE: To establish a TS model in rats secondary to tracheal autotransplantation with a graft submerged in bleomycin (Atx-Bleo). Additionally, to evaluate the clinical and histological changes, as well as the expression of newly formed collagen (NFC), isoforms of transforming growth factor beta (TGFß), fibronectin (FN), elastin (ELN), integrin ß1 (ITGß1), and matrix metalloproteinase 1 (MMP1) in TS. METHODS: Twenty Wistar rats were divided into three groups: group I (n = 20) control; group II (n = 10) end-to-end anastomosis of the trachea (tracheoplasty); and group III (n = 10) Atx-Bleo. The animals were evaluated clinically, tomographically, macroscopically, morphometrically, and microscopically. NFC deposition, and the expression of profibrotic and antifibrotic proteins were evaluated in tracheal scars. RESULTS: All animals survived the surgical procedure and the study period. Compared with the other study groups, the Atx-Bleo group developed TS and fibrosis, exhibited higher expression of NFC, TGFß1, TGFß2, FN, ELN, and ITGß1, and mild expression of TGFß3 and MMP1 (p < 0.005; analysis of variance, Dunnett and Tukey tests). CONCLUSION: Atx-Bleo in TS model rats produces tomographic and histological changes, and induces the upregulation of profibrotic proteins (TGFß1, TGFß2, collagen, FN, ELN, ITGß1) and downregulation of antifibrotic proteins (TGFß3, MMP1). Therefore, this model may be used to test new pharmacological treatments for reversing or preventing TS, and conduct basic studies regarding its pathophysiology.
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Estenose Traqueal , Animais , Colágeno/metabolismo , Matriz Extracelular , Proteínas da Matriz Extracelular/metabolismo , Metaloproteinase 1 da Matriz/metabolismo , Ratos , Ratos Wistar , Traqueia/metabolismo , Traqueia/patologia , Traqueia/cirurgia , Estenose Traqueal/etiologia , Estenose Traqueal/patologia , Estenose Traqueal/cirurgia , Transplante AutólogoRESUMO
Lung transplantation requires optimization of donor's organ use through ex vivo lung perfusion (EVLP) to avoid primary graft dysfunction. Biomarkers can aid in organ selection by providing early evidence of suboptimal lungs during EVLP and thus avoid high-risk transplantations. However, predictive biomarkers of pulmonary graft function such as endothelin-converting enzyme (ECE-1) and vascular endothelial growth factor (VEGF) have not been described under EVLP with standard prolonged hypothermic preservation, which are relevant in situations where lung procurement is difficult or far from the transplantation site. Therefore, this study is aimed at quantifying ECE-1 and VEGF, as well as determining their association with hemodynamic, gasometric, and mechanical ventilatory parameters in a swine model of EVLP with standard prolonged hypothermic preservation. Using a protocol with either immediate (I-) or delayed (D-) initiation of EVLP, ECE-1 levels over time were found to remain constant in both study groups (p > 0.05 RM-ANOVA), while the VEGF protein was higher after prolonged preservation, but it decreased throughout EVLP (p > 0.05 RM-ANOVA). Likewise, hemodynamic, gasometric, mechanical ventilatory, and histological parameters had a tendency to better results after 12 hours of hypothermic preservation in the delayed infusion group.
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Enzimas Conversoras de Endotelina/análise , Circulação Extracorpórea/métodos , Hipotermia Induzida , Fator A de Crescimento do Endotélio Vascular/análise , Animais , Biomarcadores/análise , Hipotermia Induzida/métodos , Pulmão/fisiologia , Pulmão/cirurgia , Transplante de Pulmão , Preservação de Órgãos/métodos , Suínos , Fatores de TempoRESUMO
OBJECTIVE: To assess the presence of CLDN4 in bronchoalveolar lavage fluid (BALF) and pulmonary tissue as an early indicator of LIRI and its relationship with changes in pulmonary physiology, edema formation and histology in an experimental porcine model of LTx with CIT of 50 min or 6 h. METHODS: In 12 pigs, LIRI was produced by: group I (n = 6) LTx with 50 min of CIT (LTx-50 min-CIT); and group II (n = 6) LTx with 6 h of CIT (LTx-6h-CIT). The lung function, edema formation, macroscopic and microscopic changes were assessed. CLDN4 expression in BALF and pulmonary tissue were determined. RESULTS: Both groups presented similar clinical, edema, and histological damage, as well as similar expression of CLDN4 in BALF and tissue (p > 0.05, RM-ANOVA). CONCLUSION: CLDN4 expressed in BALF and the pulmonary tissue during the first 5 h within 72 h of the PGD window are not associated by the deterioration of lung function, edema and lung histological injury, in LTx with CIT 50 min or 6 h, CLDN4 does not seem to be a valuable indicator of LIRI.
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Claudina-4/metabolismo , Transplante de Pulmão , Traumatismo por Reperfusão , Animais , Líquido da Lavagem Broncoalveolar , Pulmão , Transplante de Pulmão/efeitos adversos , Traumatismo por Reperfusão/etiologia , SuínosRESUMO
BACKGROUND: Peripheral parenteral nutrition (PPN) is increasingly considered as an alternative to central parenteral nutrition (CPN) given the higher cost and more frequent clinical complications associated with the latter. However, the assessment of potential risks and benefits of PPN in critically ill pediatric canine patients has not been extensively performed. In this study, we aimed to explore the effect of short-term, hypocaloric PPN on weight loss, length of hospital stay, the incidence of complications, adverse effects, and mortality in critically ill pediatric canine patients. RESULTS: Between August 2015 and August 2018, a total of 59 critically ill pediatric canine patients aged from 1 to 6 months admitted at the Veterinary Sciences Research Institute of the Autonomous University of Baja California were included in this non-randomized clinical trial. Canine pediatric patients were initially allocated to 3 groups: 11 in group 1 receiving parenteral nutrition (PN) supplementation equivalent to 40% of the resting energy requirement (RER), 12 in group 2 receiving supplementation of 50% of the RER, and 36 in group 3 receiving no PN supplementation. After establishing that there was no significant difference between 40 and 50% of PN supplementation, these groups were not separated for downstream analysis. Similar lengths of hospital stays were noted among study subjects who received PN supplementation and those who did not (4.3 ± 1.5 vs. 5.0 ± 1.5, days, p = 0.097). No metabolic-, sepsis- or phlebitis-related complications were observed in any animal in the PPN supplemented group. Higher mortality (19.4% vs. 0%, p = 0.036), and a greater percentage of weight loss (9.24% vs. 0%, p < 0.001) were observed in patients who received no supplementation. CONCLUSION: Even though short-term, hypocaloric PPN did not reduce the length of hospital stay, it was associated with lower mortality and resulted in mitigation of weight loss. In contrast to previous studies evaluating central and peripheral parenteral nutrition protocols, we observed a lower frequency of metabolic, septic, and phlebitis complications using a 40-50% parenteral nutrition treatment. The parenteral nutrition therapeutic intervention used in our study may reduce PN-related adverse effects and promote a favorable disease outcome in critically ill canine patients. Larger studies will be needed to confirm these observations.
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Distance learning is a rapidly spreading form of education worldwide and it plays a crucial role to provide access to millions of people in developing countries. However, the benefits of online learning extend far beyond and became increasingly popular also in medical schools, including veterinary medicine [1,2]. In these new teaching settings, high-quality, validated and easily accessible multimedia materials are of great importance, especially in specific fields, such as radiology, where graphic resources can greatly support learning [3]. The dataset presented along with this article reviews examples of 153 canine latero-lateral thoracic radiographic medical images taken and collected at the Small Animal Veterinary Teaching Hospital, Veterinary Sciences Research Institute, Autonomous University of Baja California. serves as a basis for teaching VHS calculation. Images on this dataset contain all relevant anatomical structures in the determination of VHS, which makes them optimal images for practice in calculating VHS and teaching this procedure. The number of cardiothoracic radiographic images presented here can be a great support in learning the calculation of VHS, especially when combined with distance competency-based educators' support. Acquired heart diseases in the dog are very common, clinical evidence of degenerative valvular disease is detected in approximately 30% of dogs aged 13 years and older [4]. The prevalence of Dilated Cardiomyopathy (DCM) is remarkably high in certain breeds, approximately 25% of Irish Wolfhounds, 33% of female Doberman Pinschers, and 50% of male Doberman Pinschers are diagnosed with DCM [5]. Thoracic radiography is a key component in cardiovascular evaluation, which is used to calculate the vertebral heart score (VHS). VHS measurement has been described as one of the most objective methods for assessing cardiomegaly in dogs. VHS, in addition to thorough patient history and physical examination, can be very efficient to raise clinical suspicion of underlying heart disease. The measurement is based on cardiac height and width and is normalized to overall body size by comparison to vertebral body length. ln addition to the initial assessment of heart size, the VHS method is also useful for monitoring changes in heart size in response to treatment or to assess the progression of cardiomegaly over time in canine patients [6]Fig. 1. Importantly, considerable breed and individual variation exist with VHS and dogs can have values that fall outside of the normal range even without any cardiac disease. Thus, while the VHS method is a very informative tool, it should not be used as the only means of diagnosing cardiac disease in any given patient [7].
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Treatment of tracheal stenosis is occasionally performed in combination with wound healing modulators to manipulate new extracellular matrix (ECM) formation and prevent fibrosis. Hyaluronic acid (HA) and collagen-polyvinylpyrrolidone (collagen-PVP) decrease fibrosis in experimental tracheal healing. However, they have not been used clinically as their effect on ECM components, which modify tracheal scarring, has not been described. Objective. To evaluate the effect of the application of HA, collagen-PVP, a mixture of HA and collagen-PVP (HA+collagen-PVP), and mitomycin C on the expression of decorin, matrix metalloproteinase 1 (MMP1), and MMP9, as well as the type of collagen and deposits formed in the scar after resection and end-to-end anastomosis (REEA) of the cervical trachea using an experimental model. Materials and Methods. Thirty dogs underwent REEA of the cervical trachea and were treated with different wound healing modulators: group I (n = 6), control; group II (n = 6), HA; group III (n = 6), collagen-PVP; group IV (n = 6), HA+collagen-PVP; and group V (n = 6), mitomycin C. The dogs were evaluated clinically and endoscopically for 4 weeks. Subsequently, macroscopic and microscopic changes, expression of ECM proteins, and collagen deposition in tracheal scars were analysed. Results. Groups II, III, and IV showed reduced endoscopic, macroscopic, and microscopic inflammation, improved neovascularization, high decorin expression (p < 0.01, analysis of variance (ANOVA)), and moderate expression of MMP1 (p < 0.003, ANOVA) and type I and III collagen (p < 0.05, Kruskal-Wallis). Groups IV and V developed fewer collagen deposits (p < 0.001, ANOVA). Conclusion. Treatment with HA and collagen-PVP improved post-REEA healing by increasing neovascularization, stimulating the expression of decorin, and regulating the expression of MMP1, as well as type I and III collagen and their deposition.
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Cicatriz/tratamento farmacológico , Colágeno/administração & dosagem , Ácido Hialurônico/administração & dosagem , Complicações Pós-Operatórias/tratamento farmacológico , Povidona/administração & dosagem , Estenose Traqueal/cirurgia , Anastomose Cirúrgica , Animais , Cicatriz/etiologia , Cicatriz/patologia , Colágeno/metabolismo , Decorina/metabolismo , Modelos Animais de Doenças , Cães , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/metabolismo , Matriz Extracelular/patologia , Feminino , Fibrose , Humanos , Masculino , Metaloproteinase 1 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Mitomicina/administração & dosagem , Complicações Pós-Operatórias/metabolismo , Complicações Pós-Operatórias/patologia , Traqueia/efeitos dos fármacos , Traqueia/patologia , Traqueia/cirurgia , Cicatrização/efeitos dos fármacosRESUMO
Autophagy has been involved in the pathogenesis of various lung diseases. However, it is not yet known whether autophagy plays a role in hypersensitivity pneumonitis (HP). HP is an interstitial lung disease resulting from exposure to a wide variety of antigens that provoke an exaggerated immune response in susceptible individuals. The aim of this study was to explore the localization of autophagy key proteins in lungs from HP patients and controls by immunohistochemistry and analyze their expression levels by immunoblot. Macrophages and epithelial cells were strongly positive for the autophagosome biomarker LC3B (microtubule-associated protein light chain 3 beta) in HP lungs compared with controls. A similar pattern was found for the autophagy receptor p62 and the enzyme ATG4B. Unexpectedly, nuclear p62 signal was also noticed in macrophages from HP lungs. Regarding ATG5 and ATG7 localization, we observed positive staining in neutrophils, vascular smooth muscle cells, and endothelial cells. Our findings provide for the first time evidence that proteins from the autophagy machinery are highly expressed in the lungs of HP patients and describe the specific cellular and subcellular localization of LC3B, p62, ATG4B, ATG5, and ATG7 in HP lungs.
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Alveolite Alérgica Extrínseca/metabolismo , Proteínas Relacionadas à Autofagia/metabolismo , Pulmão/metabolismo , Autofagossomos/metabolismo , Estudos de Casos e Controles , Feminino , Regulação da Expressão Gênica , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Transporte ProteicoRESUMO
BACKGROUND: Chronic hypersensitivity pneumonitis (cHP) represents a severe lung disease often evolving to fibrosis with the subsequent destruction of the lung parenchyma. There are no approved therapies with confirmed efficacy to deal with this disease. METHODS: We performed an open-label, proof of concept study, to evaluate the efficacy and safety of pirfenidone added to immunosuppressive drugs on the treatment of cHP. We included 22 patients assigned to two groups: Group 1, nine patients that received prednisone plus azathioprine and Group 2, thirteen patients, received prednisone plus azathioprine and pirfenidone (ClinicalTrials.gov identifier NCT02496182). There were no significant imbalances in clinically relevant baseline characteristics between two study groups. RESULTS: After 1 year of treatment, inclusion of pirfenidone was not associated with improved forced vital capacity (primary end-point). A not significant tendency to show higher improvement of diffusion capacity of the lung for carbon monoxide (DLCO) was observed in the group receiving pirfenidone (p=0.06). Likewise, a significant improvement in the total score on the SGRQ was found in the group 2 (p=0.02) without differences in other two questionnaires related to quality of life (ATAQ-IPF and EQ-5D-3L). HRCT showed a decrease of the ground glass attenuation without changes in the fibrotic lesions and without differences between both groups. CONCLUSIONS: These findings suggest that the addition of pirfenidone to the anti-inflammatory treatment in patients with chronic HP may improve the outcome with acceptable safety profile. However, prospective randomized double-blind, placebo-controlled trials in largest cohorts are needed to validate its efficacy.