G. E. Carpagnano

Menopausal asthma: A new biological phenotype?

To cite this article: Foschino Barbaro MP, Costa VR, Resta O, Prato R, Spanevello A, Palladino GP, Martinelli D, Carpagnano GE. Menopausal asthma: a new biological phenotype? Allergy 2010; 65: 1306–1312.

Repeated virus identification in the airways of patients with mild and severe asthma during prospective follow-up

To cite this article: Turchiarelli V, Schinkel J, Molenkamp R, Foschino Barbaro MP, Carpagnano GE, Spanevello A, Lutter R, Bel EH, Sterk PJ. Repeated virus identification in the airways of patients with mild and severe asthma during prospective follow‐up. Allergy 2011; 66: 1099–1106.

Diurnal PaCO2 tension in obese women: relationship with sleep disordered breathing.

OBJECTIVE: Several obese subjects show a wide array of respiratory disturbances during sleep due to an increased upper-airway resistance. The aim of the present study was to evaluate diurnal PaCO2 tension in nonsmoking obese women and the possible relationship of this parameter with the presence of sleep disordered breathing (SDB).DESIGN: Cross-sectional study of PaCO2 tension in obese women.PATIENTS AND METHODS: A total of 91 nonsmoking obese women (BMI ≥30 kg/m2, aged 42.8±15.7 y) were recruited and evaluated for general and anthropometric parameters, respiratory function, sleep-related symptoms, and sleep disorders of breathing.RESULTS: A total of 10 subjects (10.9%) had diurnal hypercapnia (PaCO2≥43 mmHg). Age, BMI, neck circumference, apnoea/hypopnoea index, and nocturnal desaturation (expressed as TSTSaO2<90%; TSTSaO2<90%=percentage of total sleep time with oxyhaemoglobin saturation <90%) were significantly higher in obese patients with diurnal hypercapnia, compared to normocapnic women. Moreover, hypercapnic patients had reduced forced expiratory volume in 1 s compared to normocapnic individuals. By using multiple regression analysis, the best fitting model (r=0.62, P<0.001) for predicting diurnal PaCO2 tension in the study population showed that 24.23% of the variance may be explained by TSTSaO2<90%, according to the equation: PaCO2=0.09 age+0.07 TSTSaO2<90%+33.00.CONCLUSIONS: This study suggests that severity of SDB is the most important factor in determining diurnal PaCO2 tension in apparently healthy nonsmoking obese women.

The role of obstructive sleep apnea syndrome and obesity in determining leptin in the exhaled breath condensate

Leptin plays a key role in obstructive sleep apnea syndrome (OSAS). Leptin production in human airways has been previously evaluated by measuring leptin concentration in the exhaled breath condensate and in the induced sputum. The aim was to study leptin expression in the cells of induced sputum and in exhaled breath condensate of subjects with OSAS. Moreover, leptin concentrations in the blood were measured in the same groups of subjects. We enrolled four groups of patients: (1) obese patients with OSAS (OO); (2) non-obese patients with OSAS (NOO); (3) obese patients without OSAS (ONO); and (4) non-obese subjects without OSAS (C). Leptin expression was evaluated by immunocytochemistry in the sputum cells of the enrolled subjects. The concentrations of leptin in the exhaled breath condensate and plasma were measured by using a specific enzyme immunoassay. Leptin protein expression and the percentage of macrophages and neutrophils expressing leptin were higher in the induced sputum of OO, NOO and ONO patients than in C. Leptin concentrations in the exhaled breath condensate were significantly higher in OO patients (5.12 (3.8-6.6) ng ml(-1)) than in NOO (4.1 (3.9-5.2) ng ml(-1)) and ONO (4.2 (3.6-5.0) ng ml(-1)) patients. The concentration of leptin in plasma was significantly more elevated in OO (36 (24-65.9) ng ml(-1)) than in NOO (30.2 (12.4-51.4) ng ml(-1)), whereas it was not significantly different in ONO patients. This study showed that leptin in sputum and in the exhaled breath condensate is higher in obese patients with OSAS than in obese subjects without OSAS. Moreover, different mechanisms for determining leptin concentrations in the exhaled breath condensate and the blood are suggested.

Analysis of mitochondrial DNA alteration in new phenotype ACOS

BackgroundMitochondria contain their own DNA (MtDNA) that is very sensitive to oxidative stress and as a consequence could be damaged in quantity. Oxidative stress is largely recognized to play a key role in the pathogenesis of asthma and COPD and might have a role in the new intermediate phenotype ACOS (asthma-COPD overlap syndrome). The aim of this study was to investigate MtDNA alterations, as an expression of mitochondrial dysfunction, in ACOS and to verify whether they might help in the identification of this new phenotype and in its differentiation from asthma and COPD.MethodsTen (10) ACOS according to Spanish guidelines, 13 ACOS according to GINA guidelines, 13 COPD, 14 asthmatic patients and ten normal subjects were enrolled. They further underwent a blood, induced sputum and exhaled nitric oxide collection. Content of MtDNA and nuclear DNA (nDNA) were measured in the blood cells of patients by Real Time PCR.ResultsACOS patients showed an increase of MtDNA/nDNA ratio. Dividing ACOS according to guidelines, those from the Spanish showed a higher value of MtDNA/nDNA compared to those from GINA/GOLD (92.69 ± 7.31 vs 80.68 ± 4.16). Spanish ACOS presented MtDNA/nDNA ratio closer to COPD than asthma. MtDNA was higher in asthmatic, COPD, GINA and Spanish ACOS patients compared to healthy subjects (73.30 ± 4.47–137.0 ± 19.45–80.68 ± 4.16–92.69 ± 7.31 vs 65.97 ± 20.56).ConclusionWe found an increase of MtDNA/nDNA ratio in ACOS subjects that led us to conclude that there is presence of mitochondrial dysfunction in this disease, that makes it closer to COPD than to asthma. Although the MtDNA/nDNA ratio results are a useful marker for differential diagnosis from asthma, COPD and ACOS, further studies are needed to confirm the potentiality of MtDNA/nDNA ratio and to a better characterization of ACOS.

Evidence of lower oxidative stress in the air spaces of patients with reversible COPD.

The mechanism responsible for the reversibility of airflow limitation in stable chronic obstructive pulmonary disease (COPD) patients is unknown. The aim of this study is to assess the relationship between the reversibility of airflow limitation, the redox balance and the inflammatory cells in the sputum of patients with stable COPD. For this purpose we examined 15 normal healthy control subjects and 20 nonatopic COPD patients. The COPD patients were divided into two groups: reversible COPD (increase in FEV1>200 ml and/or ≥12% after 200 μg of inhaled salbutamol) or non-reversible COPD. GSH, GSSG were measured in induced sputum and blood. Protein carbonyls were evaluated by WB in sputum and IL-4 and IL-6 and TNF-α in plasma and sputum. GSH oxidation and protein oxidation were lower in reversible COPD patients than in those with no reversibility. The sputum eosinophil count was significantly higher in the reversible group than in the non-reversible group, and IL-4 concentration was higher in the same patients both in sputum and in plasma. In contrast, IL-6 and TNF-α were increased in non-reversible COPD patients in both biological samples. We conclude that airflow reversibility in COPD patients is associated with airway oxidative stress and activation of eosinophil inflammatory pattern in sputum and blood, suggesting that these patients could respond to specific pharmacological treatment.

Microsatellite alterations suggestive of organ-specific asthma and atopy in exhaled breath condensate.

Several genetic linkage studies demonstrated the existence of organ-specific disease susceptibility genes which may induce asthma and atopy (1). In particular, chromosomes 6p and 14q seem to host asthma/atopy susceptibility genes (2). Microsatellite alterations (MAs) in these regions were detected in sputum cells of asthmatics (3). The MAs can serve as a valid tool to discriminate between asthma and COPD patients (4). MAs are undetectable in nasal cytology samples from patients with allergic rhinitis as bronchial asthma MAs are specific for disease target organ (5). The possibility to analyze MAs in exhaled breath condensate (EBC) was demonstrated. The method was validated by the comparative study of the EBC DNA MAs and paired lung tissue DNA, allowing the use of this sample in lung disease genetic studies (6). The aim of the study was to investigate MAs of chromosomes 6p and 14q in EBC DNA of a group of asthmatics with atopy (AAs), a group of nonatopic asthmatics (ANAs) and a group of nonasthmatic atopics (As), and to verify whether atopy and asthma have different genetic traits. Twenty-eight consecutive AAs, 13 ANAs, 15 As and 15 healthy subjects (HS) were enrolled (Table 1). The diagnosis of asthma and the assessment of its severity were performed according to GINA. Peripheral whole blood (WB) and EBC were collected from each patient. DNA from WB and EBC was extracted by using QIAmp DNA mini Kit (Qiagen, Mican, Italy). Five fluorescence labeled markers located at chromosomes 14q (D14S258, D14S588, D14S292) and 6p (D6S2223, D6S263) were used to evaluate LOH and MI. DNA was amplified by the PCR. The samples were analyzed by capillary electrophoresis on ABI PRISM 310 (Applied Biosystems, Monza, Italy). Good quality DNA in terms of integrity and amount (mean quantity: 20 ng/ ll) was obtained from all EBC samples. None of the HS exhibited genetic alteration of the five markers tested supporting Paraskakis’ hypothesis that these markers are disease-specific (4). MAs in WB-DNA were absent. The microsatellites D14S292 and D6S2223 drive the susceptibility to atopy and asthma, respectively, other microsatellites are involved in both phenotypes. The increased number of MAs is associated with a declined lung function, severe asthma and high degree of atopy. The AAs exhibited the highest percentages of MAs at 6p and 14q. The highest number of MAs was found in the EBC DNA of AAs confirming the results obtained from sputum DNA (4). In accordance with Parashakis et al. (4) the correlation Exhaled microsatellite alterations at chromosomes 6p and 14q are suggestive of organ-specific asthma and atopy. ALLERGY Net

Study of mitochondrial DNA alteration in the exhaled breath condensate of patients affected by obstructive lung diseases.

Mitochondrial DNA (MtDNA) has been studied as an expression of oxidative stress in asthma, COPD, lung cancer and obstructive sleep apnea, but it has been mainly investigated systemically, although the pathogenetic mechanisms begin in the airways and only later progress to systemic circulation. The aim of this study was to investigate the MtDNA alterations in the exhaled breath condensate (EBC) of patients with asthma, COPD and asthma-COPD overlap syndrome (ACOS). In order to analyze better what happens to mitochondria, both locally and systemically, we compared MtDNA/nDNA in blood and EBC of paired patients. Thirteen (13) COPD patients, 14 asthmatics, 23 ACOS (10 according to Spanish guidelines, 13 in line with GINA guidelines) and 12 healthy subjects were enrolled. Patients underwent clinical and functional diagnostic tests as foreseen by the guidelines. They underwent blood and EBC collection. Content of MtDNA and nuclear DNA (nDNA) was measured in the blood cells and EBC of patients by Real Time PCR. The ratio between MtDNA/nDNA was calculated. For the first time we were able to detect MtDNA/nDNA in the EBC. We found higher exhaled MtDNA/nDNA in COPD, asthmatic and ACOS patients respectively compared to healthy subjects (21.9  ±  4.9 versus 6.51  ±  0.21, p  <  0.05; 7.9  ±  2.5 versus 6.51  ±  0.21, p  =  0.06; 18.3  ±  3.4 versus 6.51  ±  0.21, p  <  0.05). The level of exhaled MtDNA/nDNA was positively correlated with the plasmatic one. The levels of MtDNA/nDNA in the EBC, as expression of oxidative stress, are increased in COPD, asthmatic and ACOS patients compared to healthy subjects. These are preliminary results in a small number of well characterized patients that requires confirmation on a larger population. We support new studies directed toward the analysis of exhaled MtDNA/nDNA as a new exhaled non-invasive marker in other inflammatory/oxidative airways diseases.

Big atria, little P waves: unexpected mismatch between chamber volume and electrocardiogram signal in "false" atrial fibrillation.

We report the case of an 81-year-old woman referred for dizziness andsuspectedatrial fibrillationandhospitalizedfor bronchopneumonia and fever. The woman was diabetic in treatment with insulin; she also had history of chronic obstructive pulmonary disease, gastrointestinal bleeding after treatment with warfarin, and hypothyroidism.

Exhaled Nitric Oxide and Exhaled Breath Temperature as Potential Biomarkers in Patients with Pulmonary Hypertension

Background Pulmonary hypertension (PH) is a progressive fatal disease thus, noninvasive prognostic tools are needed to follow these patients. The aim of our study was to evaluate fractional exhaled nitric oxide (FeNO) and exhaled breath temperature (EBT) values in patients with PH from different causes and to correlate them with respiratory functional data. Methods Twenty-four PH patients underwent spirometry, carbon monoxide diffusion (DLCO) test, transthoracic echocardiography, right-heart catheterization, and FeNO and EBT measurements. Results We studied 3 groups according to the type of PH: 10 patients with pulmonary arterial hypertension (PAH) (group A), 11 patients with PH due to chronic obstructive pulmonary disease (COPD) (group B), and 3 patients with PH associated with left heart disease (group C). Mean FeNO values tend to be higher in group B (15.0 ± 9.3ppb) compared with other groups (respectively, 9.9 ± 5.7 and 8.5 ± 5.2 ppb in groups A and C; p = 0.271) but no statistical significance has been reached. Mean values of alveolar NO concentration (CANO) were higher in groups A and B compared to group C (respectively, 16.9 ± 12.6; 13.9 ± 6.8; and 6.7 ± 2.0 ppb) (p = 0.045). EBT mean values were significantly lower in group C when compared with other groups (group C: 29.0 +- 1.3°C, groups A and B: 30.9 ± 1.3 and 31.2 ± 1.2°C, respectively: p = 0.041). EBT levels were inversely correlated to mean pulmonary artery pressure (PAPm) levels (Spearman coefficient -0.481; p = 0.017). Conclusions eNO, CANO, and EBT have been evaluated in three groups of PH patients. Interestingly EBT reduction was correlated with PAPm increase, whereas FeNO was higher in COPD patients and CANO in PAH and COPD groups. Further studies are needed to clarify EBT, FeNO, and CANO roles as biomarkers in the monitoring of patients with PH.