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Biomedical subjects

Emiel F M Wouters

Publications and source records attributed to Emiel F M Wouters.

At least 19 recordsLinked to original sources
PubMed2006

Myogenic differentiation during regrowth of atrophied skeletal muscle is associated with inactivation of GSK-3beta.

Muscle atrophy contributes to morbidity and mortality in aging and chronic disease, emphasizing the need to gain understanding of the mechanisms involved in muscle atrophy and (re)growth. We hypothesized that the magnitude of muscle regrowth during recovery from atrophy determines whether myonuclear accretion and myogenic differentiation are required and that insulin-like growth factor (IGF)-I/Akt/glycogen synthase kinase (GSK)-3beta signaling differs between regrowth responses. To address this hypothesis we subjected mice to hindlimb suspension (HS) to induce atrophy of soleus (-40%) and plantaris (-27%) muscle. Reloading-induced muscle regrowth was complete after 14 days and involved an increase in IGF-IEa mRNA expression that coincided with Akt phosphorylation in both muscles. In contrast, phosphorylation and inactivation of GSK-3beta were observed during soleus regrowth only. Furthermore, soleus but not plantaris regrowth involved muscle regeneration based on a transient increase in expression of histone 3.2 and myosin heavy chain-perinatal, which are markers of myoblast proliferation and differentiation, and a strong induction of muscle regulatory factor (MRF) expression. Experiments in cultured muscle cells showed that IGF-I-induced MRF expression is facilitated by inactivation of GSK-3beta and selectively occurs in the myoblast population. This study suggests that induction of IGF-I expression and Akt phosphorylation during recovery from muscle atrophy is independent of the magnitude of muscle regrowth. Moreover, our data demonstrate for the first time that the regenerative response characterized by myoblast proliferation, differentiation, and increased MRF expression in recovering muscle is associated with the magnitude of regrowth and may be regulated by inactivation of GSK-3beta.

Animals↗
PubMed2006

Increased granzyme A expression in type II pneumocytes of patients with severe chronic obstructive pulmonary disease.

RATIONALE: Chronic obstructive pulmonary disease (COPD) is associated with increased numbers of CD8(+) cytotoxic T lymphocytes (CTLs) in the lung, but the functional activity of CTLs remains unknown. Granzyme A (GrA) and B (GrB) are serine proteases considered to be important effector molecules of CTLs and natural killer cells. OBJECTIVE: To investigate protein and mRNA expression of GrA and GrB in peripheral lung tissue from patients with COPD and control subjects with normal lung function. METHODS: Paraffin-embedded sections of surgical lung specimens from 22 patients with COPD (FEV(1), 22% predicted; GOLD stage 4) and 15 control subjects (FEV(1), 108% predicted) were immunostained for GrA and GrB, and semiquantified on a 3-point scale. Messenger RNA expression in total lung, specific cell types enriched for by laser capture microdissection, and freshly isolated primary cells were determined by reverse transcriptase-polymerase chain reaction. MEASUREMENTS AND MAIN RESULTS: GrA and GrB immunoreactivity was observed in CD8(+) CTLs and CD57(+) natural killer cells, but also in type II pneumocytes and alveolar macrophages in both groups. Bronchiolar epithelium stained positive for GrA, but negative for GrB. These observations were confirmed by reverse transcriptase-polymerase chain reaction on total lung, laser capture microdissection-enriched specific cell types and freshly isolated primary type II pneumocytes. The scores of GrA-expressing type II pneumocytes were significantly higher in patients with COPD versus control subjects. CONCLUSIONS: GrA and GrB mRNA and protein are detectable in human lung tissue. GrA expression is increased in type II pneumocytes of patients with very severe COPD. These results indicate that GrA may be important in the development of COPD.

Bronchi↗
PubMed2006

Modulation of glutaredoxin-1 expression in a mouse model of allergic airway disease.

Glutaredoxins (GRX) are antioxidant enzymes that preferentially catalyze the reduction of protein-glutathione mixed disulfides. The formation of mixed disulfides with GSH is known as S-glutathionylation, a post-translational modification that is emerging as an important mode of redox signaling. Since asthma is a disease that is associated with increased oxidative stress and altered antioxidant defenses, we investigated the expression of GRX in a murine model of allergic airway disease. Sensitization and challenge of C57BL/6 mice with ovalbumin resulted in increased expression of GRX1 mRNA, as well as increased amounts of GRX1 protein and total GRX activity in the lung. Because GRX1 expression is prominent in bronchial epithelium, we isolated primary epithelial cells from mouse trachea to investigate the presence of GRX. Primary tracheal epithelial cells were found to express both GRX1 and 2 mRNA and detectable GRX activity. Treatment with IFN-gamma increased the expression of GRX1 and overall GRX activity, resulting in attenuation of protein S-glutathionylation. In contrast, TGF-beta1 caused decreased GRX1 expression and overall GRX activity, leading to markedly enhanced protein S-glutathionylation. GRX1 joins the cadre of antioxidant defenses known to be modulated during allergic airway inflammation.

Animals↗
PubMed2006

Cellular protein breakdown and systemic inflammation are unaffected by pulmonary rehabilitation in COPD.

BACKGROUND: Pulmonary rehabilitation can improve the functional capacity, but has a variable effect on the low fat-free mass (FFM) in patients with chronic obstructive pulmonary disease. HYPOTHESIS: Pulmonary rehabilitation would not affect catabolic drives such as systemic inflammation and also protein breakdown. METHODS: Patients (n = 40) were studied at the start of an 8-week in-patient pulmonary rehabilitation programme, at the end of the programme and 4 weeks later. FFM and functional capacity (quadriceps strength, handgrip strength and peak workload) were assessed. Pseudouridine (PSU) urinary excretion (cellular protein breakdown) and inflammatory status were determined. Healthy participants had a single baseline assessment (n = 18). RESULTS: PSU, (IL)-6 and soluble tumour necrosis factor (sTNF)alpha R75 were increased in patients compared with healthy participants, whereas FFM and functional capacity were reduced (all p < 0.01). PSU was inversely related to both FFM and skeletal muscle function. FFM and functional parameters increased with rehabilitation, but PSU and inflammatory status were unaffected. The gain in FFM was lost 4 weeks after the completion of rehabilitation (p < 0.01). CONCLUSION: The anabolic effect of pulmonary rehabilitation improved FFM, but it did not reverse the increased protein breakdown or systemic inflammation. Thus, on cessation of pulmonary rehabilitation the FFM gains were lost owing to a loss of anabolic drive.

Biomarkers↗
PubMed2006

Dynamic redox control of NF-kappaB through glutaredoxin-regulated S-glutathionylation of inhibitory kappaB kinase beta.

The transcription factor NF-kappaB, a central regulator of immunity, is subject to regulation by redox changes. We now report that cysteine-179 of the inhibitory kappaB kinase (IKK) beta-subunit of the IKK signalosome is a central target for oxidative inactivation by means of S-glutathionylation. S-glutathionylation of IKK-beta Cys-179 is reversed by glutaredoxin (GRX), which restores kinase activity. Conversely, GRX1 knockdown sensitizes cells to oxidative inactivation of IKK-beta and dampens TNF-alpha-induced IKK and NF-kappaB activation. Primary tracheal epithelial cells from Glrx1-deficient mice display reduced NF-kappaB DNA binding, RelA nuclear translocation, and MIP-2 (macrophage inflammatory protein 2) and keratinocyte-derived chemokine production in response to LPS. Collectively, these findings demonstrate the physiological relevance of the S-glutathionylation-GRX redox module in controlling the magnitude of activation of the NF-kappaB pathway.

Animals↗
PubMed2006

Caffeine metabolites are inhibitors of the nuclear enzyme poly(ADP-ribose)polymerase-1 at physiological concentrations.

The activity of the nuclear enzyme poly(ADP-ribose)polymerase-1 (E.C.2.4.2.30), which is highly activated by DNA strand breaks, is associated with the pathophysiology of both acute as well as chronic inflammatory diseases. PARP-1 overactivation and the subsequent extensive turnover of its substrate NAD+ put a large demand on mitochondrial ATP-production. Furthermore, due to its reported role in NF-kappaB and AP-1 mediated production of pro-inflammatory cytokines, PARP-1 is considered an interesting target in the treatment of these diseases. In this study the PARP-1 inhibiting capacity of caffeine and several metabolites as well as other (methyl)xanthines was tested using an ELISA-assay with purified human PARP-1. Caffeine itself showed only weak PARP-1 inhibiting activity, whereas the caffeine metabolites 1,7-dimethylxanthine, 3-methylxanthine and 1-methylxanthine, as well as theobromine and theophylline showed significant PARP-1 inhibiting activity. Further evaluation of these compounds in H2O2-treated A549 lung epithelial and RF24 vascular endothelial cells revealed that the decrease in NAD+-levels as well as the formation of the poly(ADP-ribose)polymer was significantly prevented by the major caffeine metabolite 1,7-dimethylxanthine. Furthermore, H2O2-induced necrosis could be prevented by a high dose of 1,7-dimethylxanthine. Finally, antioxidant effects of the methylxanthines could be ruled out with ESR and measurement of the TEAC. Concluding, caffeine metabolites are inhibitors of PARP-1 and the major caffeine metabolite 1,7-dimethylxanthine has significant PARP-1 inhibiting activity in cultured epithelial and endothelial cells at physiological concentrations. This inhibition could have important implications for nutritional treatment of acute and chronic inflammatory pathologies, like prevention of ischemia-reperfusion injury or vascular complications in diabetes.

Adenosine Diphosphate↗
PubMed2006

Muscle wasting and impaired muscle regeneration in a murine model of chronic pulmonary inflammation.

Muscle wasting and increased circulating levels of inflammatory cytokines, including TNF-alpha, are common features of chronic obstructive pulmonary disease. To investigate whether inflammation of the lung is responsible for systemic inflammation and muscle wasting, we adopted a mouse model of pulmonary inflammation resulting from directed overexpression of a TNF-alpha transgene controlled by the surfactant protein C (SP-C) promoter. Compared with wild-type mice, SP-C/TNF-alpha mice exhibited increased levels of TNF-alpha in the circulation and increased endogenous TNF-alpha expression in skeletal muscle, potentially reflecting an amplificatory response to circulating TNF-alpha. Decreased muscle and body weights observed in SP-C/TNF-alpha mice were indicative of muscle wasting. Further evaluation of the SP-C/TNF-alpha mouse musculature revealed a decreased muscle regenerative capacity, shown by attenuated myoblast proliferation and differentiation in response to reloading of disuse-atrophied muscle, which may contribute to skeletal muscle wasting. Importantly, incubation of cultured myoblasts with TNF-alpha also resulted in elevated TNF-alpha mRNA levels and inhibition of myoblast differentiation. Collectively, our results demonstrate that chronic pulmonary inflammation results in muscle wasting and impaired muscle regeneration in SP-C/TNF-alpha mice, possibly as a consequence of an amplificatory TNF-alpha expression circuit extending from the lung to skeletal muscle.

Animals↗
PubMed2006

Patterns of inflammation and the use of reversibility testing in smokers with airway complaints.

BACKGROUND: Although both smoking and respiratory complaints are very common, tools to improve diagnostic accuracy are scarce in primary care. This study aimed to reveal what inflammatory patterns prevail in clinically established diagnosis groups, and what factors are associated with eosinophilia. METHOD: Induced sputum and blood plasma of 59 primary care patients with COPD (n = 17), asthma (n = 11), chronic bronchitis (CB, n = 14) and smokers with no respiratory complaints ('healthy smokers', n = 17) were collected, as well as lung function, smoking history and clinical work-up. Patterns of inflammatory markers per clinical diagnosis and factors associated with eosinophilia were analyzed by multiple regression analyses, the differences expressed in odds ratios (OR) with 95% confidence intervals. RESULTS: Multivariately, COPD was significantly associated with raised plasma-LBP (OR 1.2 [1.04-1.37]) and sTNF-R55 in sputum (OR 1.01 [1.001-1.01]), while HS expressed significantly lowered plasma-LBP (OR 0.8 [0.72-0.95]). Asthma was characterized by higher sputum eosinophilic counts (OR 1.3 [1.05-1.54]), while CB showed a significantly higher proportion of sputum lymphocytic counts (OR 1.5 [1.12-1.9]). Sputum eosinophilia was significantly associated with reversibility after adjusting for smoking, lung function, age, gender and allergy. CONCLUSION: Patterns of inflammatory markers in a panel of blood plasma and sputum cells and mediators were discernable in clinical diagnosis groups of respiratory disease. COPD and so-called healthy smokers showed consistent opposite associations with plasma LBP, while chronic bronchitics showed relatively predominant lymphocytic inflammation compared to other diagnosis groups. Only sputum eosinophilia remained significantly associated with reversibility across the spectrum of respiratory disease in smokers with airway complaints.

Adult↗
PubMed2006

Effect of glutamate ingestion on whole-body glutamate turnover in healthy elderly and patients with chronic obstructive pulmonary disease.

OBJECTIVE: Decreased whole-body glutamate turnover is found in healthy elderly and in patients with chronic obstructive pulmonary disease (COPD). Glutamate supplementation as an option to increase whole-body glutamate turnover and, hence, glutamate availability has never been investigated. In the present study, we developed a protocol based on repeated glutamate ingestion to increase plasma glutamate concentration to a steady-state level without inducing toxic side effects and to evaluate the effect of repeated glutamate ingestion on whole-body glutamate turnover in patients with COPD and healthy elderly. METHODS: In part 1, the response of plasma glutamate concentration was determined in young healthy volunteers who repeatedly ingested a glutamate solution. The tolerance of the glutamate drink was evaluated in 26 healthy volunteers by a food tolerance questionnaire. In part 2, eight male patients with COPD and eight healthy elderly ingested the glutamate drink, an isomolar amount of a glutamine drink, or only water to test the effect on plasma glutamate concentration and whole-body glutamate turnover. RESULTS: In part 1, repeated ingestion of 30 mg of glutamate per kilogram of body weight every 20 min increased plasma glutamate concentration five-fold to steady-state level within 80 min and without any side effects. In part 2, repeated ingestion of glutamate significantly increased whole-body glutamate turnover in healthy controls and patients with COPD, although the increase was smaller in patients with COPD than in controls. CONCLUSION: We found that repeated ingestion of 30 mg of glutamate per kilogram of body weight every 20 min can increase glutamate availability in healthy elderly and patients with COPD, who are likely more dependent on external glutamate ingestion than are young adults.

Aged↗
PubMed2006

Exercise capacity before and after an 8-week multidisciplinary inpatient rehabilitation program in lung cancer patients: a pilot study.

Although lung cancer is a highly prevalent type of cancer, the effects of an inpatient multidisciplinary rehabilitation program on pulmonary function and exercise capacity have never been studied in these patients. Pulmonary function, 6-min walking distance and peak exercise capacity of 10 patients with a severely impaired pulmonary function following treatment of lung cancer were assessed in this pilot study before and after an 8-week inpatient multidisciplinary rehabilitation program. At baseline, patients had a restrictive pulmonary function and an apparent exercise intolerance (median 6-min walking distance: 63.6% predicted; median peak cycling load: 58.5% predicted). Despite the lack of change in median pulmonary function [FEV1: -0.01L, p = 0.5469], functional exercise capacity [145 m; 43.2% of the initial values, p=0.0020] and peak exercise capacity [26 W; 34.4% of the initial values, p = 0.0078] improved significantly compared to baseline. Future trials have to corroborate the present findings. Nevertheless, patients with lung cancer have a clear indication to start a comprehensive rehabilitation program following intensive treatment of their disease. In fact, based on the results of the present pilot study it appears that these patients are good candidates for pulmonary rehabilitation programs.

Aged↗
PubMed2006

In situ detection of S-glutathionylated proteins following glutaredoxin-1 catalyzed cysteine derivatization.

S-glutathionylation is rapidly emerging as an important post-translational modification, responsible for transducing oxidant signals. However, few approaches are available that allow visualization of glutathione mixed disulfides in intact cells. We describe here a glutaredoxin1-dependent cysteine derivatization and labeling approach, in order to visualize S-glutathionylation patterns in situ. Using this new method, marked S-glutathionylation was observed in epithelial cells, which was predominant at membrane ruffles. As expected, the labeling intensity was further enhanced in response to bolus oxidant treatments, or in cells overexpressing Nox1 plus its coactivators. In addition, manipulation of endogenous levels of glutaredoxin-1 via RNAi, or overexpression resulted in altered sensitivity to H2O2 induced formation of glutathione mixed disulfides. Overall, the derivatization approach described here preferentially detects S-glutathionylation and provides an important means to visualize this post-translational modification in sub-cellular compartments and to investigate its relation to normal physiology as well as pathology.

Animals↗
PubMed2006

Cost-effectiveness of self-management in asthma: a systematic review of peak flow monitoring interventions.

OBJECTIVES: It is generally accepted that home peak flow monitoring increases patients' self-management and could lead to cost savings. The aim of this review was to analyze costs and the cost-effectiveness of self-management based on peak flow monitoring interventions in asthma. METHODS: Twenty-one studies were included in this review. Data were extracted, and methodological and economic quality were assessed. These studies presented economic information regarding self-management interventions based on peak flow monitoring in asthmatics. The mean methodological quality was 4.6 (maximum 8), and the mean economic quality was 12.0 (maximum 15). RESULTS: In eighteen studies, the interventions led to net savings compared with usual care or less intensive intervention. Only three studies found the total costs to be higher in the intervention group. In thirteen of the seventeen studies that analyzed health outcomes, at least one of the reported health outcomes improved statistically significantly after the intervention. However, the methods of economic evaluation differed among the studies and were not always in line with the standard methodology. CONCLUSIONS: The interventions, costs, and outcomes were very diverse. The results emphasize the need for guidelines to increase the comparability of cost-effectiveness evaluations relating to asthma. Only then will it be possible to conclude whether interventions for asthmatics, such as self-management based on peak flow monitoring interventions, are cost-effective.

Asthma↗
PubMed2006

Systemic inflammation in COPD: is genetic susceptibility a key factor?

COPD is a multicomponent disease characterized by abnormal inflammatory response of the lungs to noxious particles that is accompanied by systemic effects like weight loss, muscle wasting, reduced functional capacity and impaired health status. A persistent low-grade systemic inflammatory response reflected by enhanced levels of acute phase proteins like C-reactive protein (CRP) and pro-inflammatory cytokines such as tumor necrosis factor (TNF)-alpha, is present in part of the COPD population. The production of inflammatory proteins is partly genetically determined. Several studies have shown that polymorphisms within genes coding for these inflammatory mediators may modulate systemic inflammatory responses. Among all of these genes, the TNF family (TNF-alpha, lymphotoxin (LT)-alph and their receptors TNF-R55 and TNF-R75), interleukin (IL)-6 and CRP gene polymorphisms are the most prominent candidates. However, large carefully designed studies in well-characterized COPD cohorts are required to unravel the exact role of genetic background in the systemic component of this disease.

C-Reactive Protein↗
PubMed2006

Metabolic effects of glutamine and glutamate ingestion in healthy subjects and in persons with chronic obstructive pulmonary disease.

BACKGROUND: Because low plasma glutamate and glutamine concentrations are often seen in chronic obstructive pulmonary disease (COPD), glutamine or glutamate supplementation may be a good option for preventing further metabolic disturbances in COPD patients. However, the metabolic effects of glutamate supplementation have never been compared with those of glutamine supplementation. OBJECTIVE: We compared the metabolic effects of repeated ingestion of glutamine and glutamate in COPD patients and in age-matched healthy control subjects. DESIGN: On 3 d separated by intervals of > or = 2 d, a protocol of primed constant and continuous infusion of [2H5]phenylalanine and [2H2]tyrosine was performed for 3 h in 8 stable male COPD patients and 8 healthy control subjects. After a 90-min tracer infusion, all subjects ingested a glutamine or glutamate drink or the same amount of water every 20 min for 80 min. Blood samples were taken at the end of the postabsorptive and ingestion periods to test for effects on plasma amino acid and substrate concentrations and whole-body protein turnover. RESULTS: Glutamate but not glutamine ingestion resulted in higher plasma ornithine concentrations than did water ingestion (P < 0.01). The change in plasma arginine, citrulline, and urea concentrations was significantly (P < 0.01) higher after glutamine ingestion than after water or glutamate ingestion. Whole-body protein turnover decreased overall, independent of the drink consumed. CONCLUSIONS: Repeated ingestion of glutamine and glutamate resulted in different effects on the plasma amino acid concentration. In both groups, ingestion of glutamine but not of glutamate increased the plasma concentrations of citrulline and arginine, substrates produced in the intestine and the liver.

Aged↗
PubMed2006

Greater whole-body myofibrillar protein breakdown in cachectic patients with chronic obstructive pulmonary disease.

BACKGROUND: Experimental studies indicate that greater skeletal muscle protein breakdown is a trigger for the cachexia that often is prevalent in chronic obstructive pulmonary disease (COPD). OBJECTIVE: We compared myofibrillar protein breakdown (MPB) with whole-body (WB) protein breakdown (PB) in 9 cachectic COPD patients [x +/- SEM forced expiratory volume in 1 s (FEV(1)): 48 +/- 4% of predicted], 7 noncachectic COPD patients (FEV(1): 53 +/- 5% of predicted), and 7 age-matched healthy control subjects, who were matched by body mass index with the noncachectic patients. DESIGN: After the subjects fasted overnight (10 h) and discontinued the maintenance medication, a primed constant and continuous infusion protocol was used to infuse L-[ring-(2)H(5)]-phenylalanine and L-[ring-(2)H(2)]-tyrosine to measure WB protein turnover and L-[(2)H(3)]-3-methylhistidine to measure WB MPB. Three arterialized venous blood samples were taken between 80 and 90 min of infusion to measure amino acid concentrations and tracer enrichments. RESULTS: Body composition, WB protein turnover, and WB MPB did not differ significantly between the noncachectic COPD and control subjects. Cachectic COPD patients had lower fat mass and fat-free mass values (both: P < 0.01) than did the noncachectic COPD patients. WB MPB was significantly (P < 0.05) higher in the cachectic COPD group (18 +/- 3 nmol . kg(-1) . min(-1)) than in the combined control and noncachectic COPD groups (10 +/- 1 nmol . kg(-1) . min(-1)), but WB protein turnover did not differ significantly between the groups. Correlations with fat-free mass were significant (P < 0.05) for plasma glutamate and branched-chain amino acids, and that for WB MPB trended toward significance (P = 0.07). CONCLUSION: Cachexia in clinically stable patients with moderate COPD is characterized by increased WB MPB, which indicates that myofibrillar protein wasting is an important target for nutritional and pharmacologic modulation.

Aged↗
PubMed2006

Approaches to improving health status in chronic obstructive pulmonary disease: one or several?

Health outcomes represent a broad group of endpoints used in clinical practice to assess the efficacy and effectiveness of interventions and to assess disease outcomes. Health status is one of the widely applied patient-reported health outcomes. The outcome of a variety of interventions, targeting local impairment or local airway inflammation, is evaluated by health status measurements; these global or summative outcomes indeed quantify the overall effect of a number of biological processes. Targeting the systemic compartment to improve health status is already widely applied in the domain of pulmonary rehabilitation. Challenging developments targeting systemic inflammation and biological processes related to systemic inflammation are particularly addressed in this review.

Administration, Inhalation↗