Markku M. Nieminen

Biomarker evidence of DNA oxidation in lung cancer patients: association of urinary 8‐hydroxy‐2′‐deoxyguanosine excretion with radiotherapy, chemotherapy, and response to treatment

Ratios of urinary 8‐hydroxy‐2′‐deoxyguanosine to urinary creatinine (8‐OHdG/creatinine) have been considered as a good biological indicator of DNA oxidation. Urinary 8‐OHdG/creatinine levels of lung cancer patients were evaluated by enzyme‐linked immunosorbent assay using a monoclonal antibody N45.1 during radiotherapy and chemotherapy. An increase in urinary 8‐OHdG/creatinine was found in non‐small‐cell carcinoma (non‐SCC) patients during the course of radiotherapy. SCC patients showed higher levels of urinary 8‐OHdG/creatinine than the controls. Furthermore, SCC patients with complete or partial response to the chemotherapy showed a significant decrease in urinary 8‐OHdG/creatinine while patients with no change or progressive disease showed an increase.

Mortality in COPD patients discharged from hospital: the role of treatment and co-morbidity

BackgroundThe aim of this study was to analyse mortality and associated risk factors, with special emphasis on health status, medications and co-morbidity, in patients with chronic obstructive pulmonary disease (COPD) that had been hospitalized for acute exacerbation.MethodsThis prospective study included 416 patients from each of the five Nordic countries that were followed for 24 months. The St. Georges Respiratory Questionnaire (SGRQ) was administered. Information on treatment and co-morbidity was obtained.ResultsDuring the follow-up 122 (29.3%) of the 416 patients died. Patients with diabetes had an increased mortality rate [HR = 2.25 (1.28–3.95)]. Other risk factors were advanced age, low FEV1 and lower health status. Patients treated with inhaled corticosteroids and/or long-acting beta-2-agonists had a lower risk of death than patients using neither of these types of treatment.ConclusionMortality was high after COPD admission, with older age, decreased lung function, lower health status and diabetes the most important risk factors. Treatment with inhaled corticosteroids and long-acting bronchodilators may be associated with lower mortality in patients with COPD.

Pulmonary distribution and clearance of two beclomethasone liposome formulations in healthy volunteers

The pulmonary distribution and clearance of 99mTc-labelled beclomethasone dipropionate (Bec) dilauroylphosphatidylcholine (DLPC) and dipalmitoylphosphatidylcholine (DPPC) liposomes were compared in 11 healthy volunteers using gamma scintigraphy. As delivered by using the Aerotech jet nebulizer both liposome aerosols had a suitable droplet size (mass median aerodynamic diameter 1.3 microm) allowing deep pulmonary deposition. However, in the total drug output during the inhalation there was a relatively large difference between DLPC and DPPC of 11.4 and 3.1 microg, respectively. In a gamma camera study no significant differences existed in the central/peripheral lung deposition between the DLPC and DPPC formulations. Progressive clearance of both Tc-labelled Bec liposomes was seen: 24 h after inhalation, 79% of the originally deposited radioactivity of DLPC liposomes and 83% of that of DPPC liposomes remained in the lungs. Thus there was slightly slower clearance of inhaled liposomes using DPPC instead of DLPC. We conclude that both liposome formulations are suitable for nebulization, although aerosol clouds were more efficiently made from the DLPC liposome suspension. Our results support the view that liposome encapsulation of a drug can offer sustained release and drug action in the lower airways.

Effects of anthracyclin-based chemotherapy on total plasma antioxidant capacity in small cell lung cancer patients

Plasma total peroxyl radical trapping antioxidant parameters (TRAP) and their main antioxidant components (vitamin E, uric acid, protein sulfhydryl groups, and unidentified antioxidant proportions) were measured in 12 small cell lung cancer (SCLC) patients receiving combined chemotherapy consisting of vincristine, adriamycin and cyclophosphamide for SCLC. Plasma samples were collected ten times during the first two cycles of chemotherapy. There is previous evidence that many anticancer drugs exert their cytotoxity via free oxygen radicals. We hypothesized that adriamycin-induced, increased oxygen free radical production should decrease plasma TRAP as a consequence of oxidative stress. A statistically significant reduction of plasma TRAP was noted 8 hours after the first adriamycin infusion. A reduction of calculated TRAP (TRAPcalc)--the sum of concentrations of individual antioxidants, corrected by their experimentally-determined stochiometric factors--appeared 3 hours after the first adriamycin infusion and continued for up to 1 week afterwards. This decrease was due to the reduction of ascorbate and urate concentrations. Total TRAP, however, recovered to initial levels after 200 hours, due to an increase in unidentified antioxidants. The second course of adriamycin. These results are in accordance with previous studies showing the formation of oxidants with the use of anthracyclines. Evidence suggests that the as yet unidentified component of TRAP (UNID) increases during the oxidative stress caused by anthracycline based chemotherapy.

No relation between apolipoprotein E alleles and obstructive sleep apnea.

Apolipoprotein E (ApoE) is a genetic risk factor influencing the development of cardiovascular diseases and Alzheimers disease. Patients with obstructive sleep apnea (OSA) suffer an excess mortality and morbidity from cardiovascular diseases. The frequencies of ApoE alleles were determined in 291 patients with OSA and 728 controls. The distribution of ApoE alleles and genotypes showed no difference between OSA and controls.

Measurement of Respiratory Muscle Forces Based on Maximal Inspiratory and Expiratory Pressures

We measured the maximal inspiratory and expiratory pressures of 200 healthy subjects (94 men and 106 women) using a membrane manometer designed by the authors. The reference intervals for maximal inspiratory pressure in men were -5 to -15 kPa, and for maximal expiratory pressure 9-21 kPa. For women the reference intervals for maximal inspiratory pressure were -3 to -14 kPa and for maximal expiratory pressure 5-15 kPa. In clinical practice only the lower limits are significant. The differences between subjects of different ages were small, thus age has little bearing on the strength of respiratory muscles in healthy men and women.

Autoantibodies against Oxidised Low-Density Lipoprotein in Patients with Obstructive Sleep Apnoea

Abstract Autoantibodies against oxidised low-density lipoprotein (OxLDL-Abs) have been proposed to be an indicator of endothelial dysfunction and a novel tool for finding individuals with a high cardiovascular risk. In a cross-sectional study, OxLDL-Abs were measured in 297 patients with obstructive sleep apnoea (OSA) and 54 controls using an enzyme-linked immunosorbent assay. The autoantibodies were increased in patients with OSA when compared to controls (age, body mass index (BMI) and gender adjusted, p = 0.001). However, within the OSA patients, OxLDL-Abs were not related to smoking, hypertension or BMI, and there was a weak negative correlation (r = −0.16, P = 0.007) between age and levels of OxLDL-Abs. In conclusion, at present the measurement OxLDL-Abs still remains a method for basic research and is not applicable for screening of at-risk patients with OSA.

Easyhaler® a Novel Multiple Dose Powder Inhaler: Clinically Equivalent to Salbutamol Metered Dose Inhaler and Easier to Use

Twenty-one adult asthmatic patients participated in a trial to compare the clinical equivalence of a single dose of salbutamol inhaled either from a novel multiple dose powder inhaler (MDPI), Easyhaler, or from a conventional metered dose inhaler (MDI). The trial was carried out as a randomized, double-blind, crossover study. The study involved 2 study days with a 6-hour follow-up period of spirometric indices. In addition, blood pressure and heart rate were measured immediately before each lung function test. Our data indicate that salbutamol treatment with the MDPI achieves values which are equivalent to those achieved with the conventional pressurized MDI as regards improving pulmonary function and tolerability. The mean maximum forced expiratory volume in 1 s (FEV1) after the powder dose was 2.44 +/- 0.96 liters and after the aerosol dose 2.45 +/- 0.93 liters. The mean area under the curve of absolute FEV1 values was 822 +/- 340 and 829 +/- 335, respectively. The mean percent change from the baseline in FEV1, forced vital capacity and peak expiratory flow following administration of the preparations was of equal magnitude in both cases. The treatments tested had no effect on blood pressure or heart rate and were well tolerated. A further important finding was that most patients found the MDPI easier or no more difficult to use than the conventional MDI and this probably facilitates the transition from pressurized MDIs to the novel MDPI.

A rapid dosimetric methacholine challenge in asthma diagnostics: a clinical study of 230 patients with dyspnoea, wheezing or a cough of unknown cause

The rapid methacholine challenge test using a pocket turbine spirometer (Micro Spirometer) and the Spira Elektro 2 dosimeter was performed with 230 consecutive patients who had dyspnoea, wheezing or a prolonged cough of unknown cause. Patients with previous asthma diagnoses as well as those who had used inhaled steroids during the preceding 4 weeks were excluded. Seventy-eight patients (34%) were methacholine positive (PD20FEV1 < or = 6900 micrograms) 47 (60%) of whom had a final diagnosis of American Thoracic Society (ATS) criteria fulfilling bronchial asthma. One hundred and fifty-two patients (66%) were methacholine negative (PD20FEV1 > 6900 micrograms) 14 (9%) of whom had bronchial asthma according to clinical evaluation. Increased bronchial responsiveness was strongly associated with ATS criteria fulfilling asthma (P < 0.0001). When PD20FEV1 was used, 47 (77%) of the asthmatic patients were hyper-responsive (range 40-6900 micrograms) compared to 31 (18%) of the non-asthmatic patients (range 160-6900 micrograms). When using PD15FEV1, 51 (84%) of the asthmatic patients (range 28-6900 micrograms) and 52 (31%) of the non-asthmatic patients (range 100-6900 micrograms) were hyper-responsive. The level of bronchial responsiveness measured by both PD20FEV1 and PD15FEV1 differed significantly between asthmatic and non-asthmatic patients (P < 0.0001). Hyper-responsiveness was associated with an increased daily variation in peak expiratory flow (PEF) (P < 0.0001) and an increased number of blood eosinophils (P < 0.0001). Hyper-responsiveness was also associated with decreased levels of FEV1 and percentages of predicted FEV1 (P = 0.04 and P < 0.0001, respectively). Stepwise logistic regression analysis showed that the number of positive prick results (OR = 1.15, 95% CI 1.01-1.31), blood eosinophils (1.004, 1.00-1.01), level of FEV1 (0.56, 0.36-0.87) and current smoking (2.36, 1.00-5.59) were factors significantly associated with the probability of hyper-responsiveness. Age, gender, atopy, pets and a history of ex-smoking were not significantly associated with hyper-responsiveness, neither in univariate nor in multivariate analyses. The Bayesian analysis was used to investigate the diagnostic value of the rapid methacholine challenge test. A receiver operator characteristic curve demonstrated that PD20FEV1 separated asthmatic and non-asthmatic patients better than PD15FEV1. The best cutoff value of PD20FEV1 was 6000 micrograms, but the difference from 6900 micrograms was minimal. The best results of the test using a PD20FEV1 cutoff point of 6900 micrograms (PPV: 0.80, NPV: 0.79) were obtained when the pre-test probability was 0.48. The interval security of the test was established by a pre-test probability between 0.19 and 0.78. Maximal positive (0.34) and negative (0.31) final gains were achieved when pre-test probabilities were 0.33 and 0.65, respectively. The cutoff level of 150 micrograms gave 100% of specificity and predictive value of a positive test for clinical asthma diagnosis. The Bayesian analysis approach demonstrated that the test is useful in asthma diagnostics if not performed on patients with lowest or highest probabilities of asthma.

Association between apolipoprotein E alleles and autoantibodies against oxidised low-density lipoprotein.

Sir, Plasma lipoproteins from apolipoprotein E (ApoE) – deficient mice are more susceptible to in vitro oxidation than the lipoproteins from wild-type mice (1). It has been proposed that the isoforms of ApoE possess antioxidant activity in vitro with an order of efficacy E 2 > E 3 > E 4 (2). We have recently reported that autoantibodies against oxidised low-density lipoprotein (OxLDL-Ab) are elevated in patients with obstructive sleep apnoea (OSA) when compared to healthy controls (3). However, there was a considerable overlap between the patients and controls, and no relation between the autoantibody levels and smoking, hypertension or body mass index. Therefore we evaluated further the material to see whether the ApoE phenotypes and OxLDL-Ab levels are related. The 297 (27 females) OSA patients, with a mean age of 53.3 years (range 26–75) (3), sleep studies (3), enzyme-linked immunosorbent assay for measuring OxLDL-Ab (3) and ApoE phenotyping (4) have been described earlier. Sixty-six (23 females) hospital employees or orthopaedic outpatients, with a mean age of 45.6 years (24 63) served as healthy controls. The antibody titres were calculated as mean optical density values from duplicate measurements. Binding to native LDL (natLDL) was adopted as a non-specific control. The data are presented as the absolute value for the ratio of antibody binding to OxLDL/natLDL. Nine of the healthy subjects belonged to ApoE 2 phenotypes (1 E 2/2, 8 E 2/3), 38 to ApoE 3/3 phenotype and 18 to ApoE 4 phenotypes (16 E 3/4, 2 E 4/4). Among the OSA patients, 27 had one of the ApoE 2 phenotypes (E 2/3), 170 the ApoE 3/3 phenotype and 96 belonged to the ApoE 4 phenotypes (79 E 3/4, 17 E 4/4). One healthy subject and four OSA patients with the ApoE 2/4 phenotype were left out of the comparison. The healthy subjects with ApoE 2 phenotypes had a lower level of OxLDL-Ab than the subjects with other phenotypes (Figure 1). In contrast, bearing either an ApoE ε 2 or ε 4 allele was not related to the level of OxLDL-Ab in the OSA patients. The difference in OxLDL-Ab between female and male healthy subjects, mean (SD) 1.88 (0.43) and 1.67 (0.37), respectively, was significant (p = 0.05). However, within the OSA group there were no gender distinctions. The results did not reveal an allele specific antioxidant activity of ApoE isoforms as suggested by Miyata and Smith (2) except that nine healthy carriers of ApoE ε 2 had a lower level of OxLDL-Ab. We evaluated further this group of healthy subjects to see whether the OxLDL-Ab levels were related to total cholesterol, triglycerides, high density lipoprotein or fasting plasma glucose: there was a relation between the autoantibody levels and triglycerides (r = –0.270, p = 0.035) and total cholesterol (r = –0.242, p = 0.058). After adjusting by cholesterol and triglycerides, the difference in OxLDLAb levels between ApoE 2 and E 3 and E 4 phenotypes was still significant (ANCOVA, p = 0.001 and 0.008, respectively). These results suggest that ApoE 2 may have a favourable antioxidative effect in a population without major cardiovascular risk factors. However, in a crosssectional sample from a population with several risk factors capable to cause oxidative stress, such as smoking, obesity related hyperlipidaemia and OSA it-