Eosinophil and T Cell Markers Predict Functional Decline in COPD Patients

BACKGROUND. The major marker utilized to monitor COPD patients is forced expiratory volume in one second (FEV1). However, asingle measurement of FEV1 cannot reliably predict subsequent decline. Recent studies indicate that T lymphocytes and eosinophils are important determinants of disease stability in COPD. We therefore measured cytokine levels in the lung lavage fluid and plasma of COPD patients in order to determine if the levels of T cell or eosinophil related cytokines were predictive of the future course of the disease. METHODS. Baseline lung lavage and plasma samples were collected from COPD subjects with moderately severe airway obstruction and emphysematous changes on chest CT. The study participants were former smokers who had not had a disease exacerbation within the past six months or used steroids within the past two months. Those subjects who demonstrated stable disease over the following six months (ΔFEV1 % predicted = 4.7 ± 7.2; N = 34) were retrospectively compared with study participants who experienced a rapid decline in lung function (ΔFEV1 % predicted = -16.0 ± 6.0; N = 16) during the same time period and with normal controls (N = 11). Plasma and lung lavage cytokines were measured from clinical samples using the Luminex multiplex kit which enabled the simultaneous measurement of several T cell and eosinophil related cytokines. RESULTS AND DISCUSSION. Stable COPD participants had significantly higher plasma IL-2 levels compared to participants with rapidly progressive COPD (p = 0.04). In contrast, plasma eotaxin-1 levels were significantly lower in stable COPD subjects compared to normal controls (p < 0.03). In addition, lung lavage eotaxin-1 levels were significantly higher in rapidly progressive COPD participants compared to both normal controls (p < 0.02) and stable COPD participants (p < 0.05). CONCLUSION. These findings indicate that IL-2 and eotaxin-1 levels may be important markers of disease stability in advanced emphysema patients. Prospective studies will need to confirm whether measuring IL-2 or eotaxin-1 can identify patients at risk for rapid disease progression.National Heart, Lung, and Blood Institute (NO1-HR-96140, NO1-HR-96141-001, NO1-HR-96144, NO1-HR-96143; NO1-HR-96145; NO1-HR-96142, R01HL086936-03); The Flight Attendant Medical Research Institute; the Jo-Ann F. LeBuhn Center for Chest Diseas

Metformin attenuates the exacerbation of the allergic eosinophilic inflammation in high fat-diet-induced obesity in mice

A positive relationship between obesity and asthma has been well documented. The AMP-activated protein kinase (AMPK) activator metformin reverses obesity-associated insulin resistance (IR) and inhibits different types of inflammatory responses. This study aimed to evaluate the effects of metformin on the exacerbation of allergic eosinophilic inflammation in obese mice. Male C57BL6/J mice were fed for 10 weeks with high-fat diet (HFD) to induce obesity. The cell infiltration and inflammatory markers in bronchoalveolar lavage (BAL) fluid and lung tissue were evaluated at 48 h after ovalbumin (OVA) challenge. HFD obese mice displayed peripheral IR that was fully reversed by metformin (300 mg/kg/day, two weeks). OVA-challenge resulted in higher influx of total cell and eosinophils in lung tissue of obese mice compared with lean group. As opposed, the cell number in BAL fluid of obese mice was reduced compared with lean group. Metformin significantly reduced the tissue eosinophil infiltration and prevented the reduction of cell counts in BAL fluid. In obese mice, greater levels of eotaxin, TNF-α and NOx, together with increased iNOS protein expression were observed, all of which were normalized by metformin. In addition, metformin nearly abrogated the binding of NF-κB subunit p65 to the iNOS promoter gene in lung tissue of obese mice. Lower levels of phosphorylated AMPK and its downstream target acetyl CoA carboxylase (ACC) were found in lung tissue of obese mice, which were restored by metformin. In separate experiments, the selective iNOS inhibitor aminoguanidine (20 mg/kg, 3 weeks) and the anti-TNF-α mAb (2 mg/kg) significantly attenuated the aggravation of eosinophilic inflammation in obese mice. In conclusion, metformin inhibits the TNF-α-induced inflammatory signaling and NF-κB-mediated iNOS expression in lung tissue of obese mice. Metformin may be a good pharmacological strategy to control the asthma exacerbation in obese individuals.Fundação de Amparo à Pesquisa do Estado de São Paulo, 2012/14225-

Eosinophils enhance WNT-5a and TGF-β1 genes expression in airway smooth muscle cells and promote their proliferation by increased extracellular matrix proteins production in asthma

BACKGROUND: Recent studies have suggested that eosinophils may have a direct effect on airway smooth muscle cells (ASMC), causing their proliferation in patients with asthma, but the precise mechanism of the interaction between these cells remains unknown. We propose that changes in Wnt signaling activity and extracellular matrix (ECM) production may help explain these findings. Therefore, the aim of this study was to investigate the effect of eosinophils from asthmatic and non-asthmatic subjects on Wnt-5a, transforming growth factor β1 (TGF-β1), and ECM protein (fibronectin and collagen) gene expression and ASMC proliferation. METHODS: A total of 18 subjects were involved in the study: 8 steroid-free asthma patients and 10 healthy subjects. Peripheral blood eosinophils were isolated using centrifugation and magnetic separation. An individual co-culture of eosinophils with human ASMC was prepared for each study subject. Adhesion of eosinophils to ASMC (evaluated by assaying eosinophil peroxidase activity) was determined following various incubation periods (30, 45, 60, 120, and 240 min). The expression of Wnt-5a, TGF-β1, and ECM protein genes in ASMC was measured using quantitative real-time polymerase chain reaction (PCR) after 24 h of co-culture. Proliferation of ASMC was measured using the Alamar blue method after 48 h and 72 h of co-culture with eosinophils. RESULTS: Eosinophils from asthmatic subjects demonstrated increased adhesion to ASMC compared with eosinophils from healthy subjects (p < 0.05) in vitro. The expression of Wnt-5a, TGF-β1, collagen, and fibronectin genes in ASMC was significantly higher after 24 h of co-culture with eosinophils from asthmatic subjects, while co-culture of ASMC with eosinophils from healthy subjects increased only TGF-β1 and fibronectin gene expression. ASMC proliferation was augmented after co-culture with eosinophils from asthma patients compared with co-culture with eosinophils from healthy subjects (p < 0.05). CONCLUSIONS: Eosinophils enhance Wnt-5a, TGF-β1, fibronectin, and collagen gene expression in ASMC and promote proliferation of these cells in asthma. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02648074

Liquid-liquid Equilibrium Of The Water + Citric Acid + Short Chain Alcohol + Tricaprylin System At 298.15 K

In the past few years, new processes for citric acid recovery have been studied. Among these processes, liquid-liquid extraction has had special attention. The extraction process is attractive due to the fact that, if a good solvent is used, it presents high selectivity and efficiency, it does not affect the thermal stability of products, and it requires low energy consumption. In this work, experimental data for the liquid-liquid equilibria of water-citric acid-solvent systems were obtained. The following solvents were tested: 2-butanol, 1-butanol, and mixed solvents containing those alcohols and tricaprylin. The parameters for the NRTL and UNIQUAC models were obtained by fitting the thermodynamic models to the experimental data.463546550Lintomen, L., (1999) Evaluation of the Liquid-Liquid Extraction for Recovery and Purification of Citric Acid (In Portuguese), , M.Sc. Thesis, Chemical Engineering Faculty, State University of CampinasMitchell, R.J., Arrowsmith, A., Ashton, N., Mixed solvent systems for recovery of ethanol from dilute aqueous solution by liquid- liquid extraction (1987) Biotechnol. Bioeng., 30, pp. 348-351Kertes, A.S., King, C.J., Extraction chemistry of fermentation product carboxylic acids (1986) Biotechnol. Bioeng., 28, pp. 269-282Welsh, F.W., Willians, R.E., The use of vegetable oils to recover compounds from aqueous solutions (1989) J. Chem. Technol. Biotechnol., 46, pp. 169-178Chumpitaz, L.D.A., Coutinho, L.F., Meirelles, A.J.A., Surface tension of fatty acids and triglycerides (1999) J. Am. Oil Chem. Soc., 76, pp. 1-4Silva, L.H.M., Coimbra, J.S., Meirelles, A.J.A., Equilibrium phase behavior of poly(ethylene glycol) + potassium phosphate + water two-phase systems at various ph and temperatures (1997) J. Chem. Eng. Data, 42, pp. 398-401Tiwari, R.D., Sharma, J.P., (1970) The Determination of Carboxylic Functional Groups, , Pergamon Press: New YorkLintomen, L., Pinto, R.T.P., Batista, E., Meirelles, A.J.A., MacIel, M.R.W., Liquid-liquid equilibrium of the water + citric acid + 2-butanol + sodium chloride system at 298.15 K (2000) J. Chem. Eng. Data, 45, pp. 1211-1214Batista, E., Monnerat, S., Kato, K., Stragevitch, L., Meirelles, A.J.A., Liquid-liquid equilibrium for systems of canola oil, oleic acid and short-chain alcohols (1999) J. Chem. Eng. Data, 44, pp. 1360-1364Magnussem, T., Rasmussen, P., Fredenslund, A., Unifac parameter table for prediction of liquid-liquid equilibria (1981) Ind. Eng. Chem. Process Des. Dev., 20, pp. 331-339MacEdo, E.A., Skovborg, P., Rasmussen, P., Calculation of phase equilibria for solutions of strong electrolytes in solvent-water mixtures (1990) Chem. Eng. Sci., 45, pp. 875-882Stragevitch, L., D'Avila, S.G., Application of a generalised maximum likelihood method in the reduction of multicomponent liquid-liquid equilibrium data (1997) Braz. J. Chem. Eng., 14, pp. 41-52Grinberg, A., Povimonski, D., Apelblat, A., Liquid-liquid distribution in the ternary system: Citric acid - 2-butanol - water at 298.15 K (1991) Solvent Extr. Ion Exch., 9, pp. 127-13

Liquid-liquid equilibrium of the water plus citric acid plus 2-butanol plus sodium chloride system at 298.15 K

Liquid-liquid equilibrium data for water + citric acid + 2-butanol + NaCl at three different salt concentrations (5, 10, and 15 mass %) and at 298.15 K were determined. The parameters for the NRTL and UNIQUAC models were obtained by fitting the thermodynamic models to the experimental data.4561211121

Liquid-liquid equilibrium of the water plus citric acid plus short chain alcohol plus tricaprylin system at 298.15 K

In the past few years, new processes for citric acid recovery have been studied. Among these processes, liquid-liquid extraction has had special attention. The extraction process is attractive due to the fact that, if a good solvent is used, it presents high selectivity and efficiency, it does not affect the thermal stability of products, and it requires low energy consumption. In this work, experimental data for the liquid-liquid equilibria of water-citric acid-solvent systems were obtained. The following solvents were tested: 2-butanol, 1-butanol, and mixed solvents containing those alcohols and tricaprylin. The parameters for the NRTL and UNIQUAC models were obtained by fitting the thermodynamic models to the experimental data.46354655

Obesity enhances eosinophilic inflammation in a murine model of allergic asthma

Background and purpose:Obesity is associated with deterioration in asthma outcomes. Although airways eosinophil accumulation is characteristic of lung allergic diseases, little is known about the influence of obesity on the allergic eosinophil trafficking from bone marrow to lung tissues, and recruitment to airways lumen. Here, we have assessed the effects of diet-induced obesity on allergic eosinophilic inflammation in mice, examining eosinophil trafficking from bone marrow to airways, and production of T(H)1/T(H)2 cytokines.Experimental approach:C57BL/6 mice fed for 10 weeks with standard chow or high-fat diet were sensitized and challenged with ovalbumin. At 24-96 h post-ovalbumin challenge, bronchoalveolar lavage (BAL) fluid, lung tissue and bone marrow were examined.Key results:The high-fat-fed mice exhibited increased body weight and epididymal fat, glucose intolerance and alterations in lipid profile compared with the lean mice. Obesity markedly elevated serum leptin and lowered adiponectin levels. Ovalbumin challenge in obese mice promoted a markedly higher eosinophil accumulation in bone marrow and connective tissue surrounding the bronchial and bronchiolar segments. Eosinophil number in BAL fluid of obese mice was lower at 24 and 48 h. Levels of interleukin (IL)-5, eotaxin, tumour necrosis factor-alpha and IL-10 in BAL fluid of obese mice were significantly higher than in lean mice.Conclusions and implications:Diet-induced obesity enhanced eosinophil trafficking from bone marrow to lung tissues, and delayed their transit through the airway epithelium into the airway lumen. Consequently, eosinophils remain longer in lung peribronchiolar segments due to overproduction of T(H)1/T(H)2 cytokines and chemokines.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

High-fat diet associated with obesity induces impairment of mouse corpus cavernosum responses

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOWhat's known on the subject? and What does the study add? Erectile dysfunction (ED) is defined as the persistent inability to achieve and/or maintain an erection sufficient for satisfactory sexual performance. ED has been found in patients with cardiovascular and endocrine-metabolic diseases. Overweight, obesity and weight gain have been shown to be independent risk factors for the development of ED. Clinical studies show that ED should be considered an early clinical manifestation of risk factors for cardiovascular events, including acute myocardial infarction. However, the mechanisms that explains ED associated with obesity are yet to be fully elucidated. Using a mice model of high-fat diet associated with obesity, we have demonstrated that ED is the result of impaired endothelial and nitrergic cavernosal relaxations along with increased contractile responses, favouring penile detumescence. OBJECTIVE Obesity induced by high-fat diet (HFD) is one of the most important risk factor for the development of erectile dysfunction (ED) in man. This study aimed to characterize the ED resulting from obesity associated with HFD in mice. MATERIALS AND METHODS C57BL/6 mice fed for 10 weeks with either HFD to induce obesity or a standard-chow diet (SD) were used. Corpus cavernosum was surgically dissected free, and strips were mounted in 10-mL organ baths containing Krebs solution. Functional responses to endothelium-dependent and -independent agents, as well as to electrical-field stimulation were measured in the cavernosal tissue. Levels of cGMP in erectile tissue were detected by enzyme immunoassay assay. RESULTS The potency (pEC(50)) and maximal response (E(max)) to acetylcholine were significantly lower in the HFD group compared with the SD group. A marked decrease in the non-adrenergic non-cholinergic (nitrergic) cavernosal relaxations in the HFD group was also detected. There were no significant differences between the SD and HFD groups for the cavernosal relaxations in response to sodium nitroprusside. The contractile responses elicited by the alpha(1)-adrenoceptor agonist phenylephrine were significantly greater in the HFD group compared with the SD group. Similarly, the electrical-field stimulation (2-8 Hz)-induced adrenergic contractions were markedly greater in HFD mice. The pEC(50) for endothelin-1 was about 6.9-fold higher in the HFD compared with SD group. The basal cGMP content was 47% lower in HFD strips compared with SD group. There were no morphological alterations in erectile tissue of HFD group compared with SD mice. CONCLUSION Obesity associated with HFD favours ED as result of impaired endothelial and nitrergic cavernosal relaxations along with increased contractile responses to adrenergic stimulation and endothelin-1 receptor activation1071016281634FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOsem informaçã