Immanuels Taivans

Decreased FOXP3 expression in small airways of smokers with COPD

CD4+CD25+ FOXP3-positive T-regulatory cells have an important role in controlling immune and inflammatory reactions. The present authors hypothesise that these cells may be involved in the pathogenesis of chronic obstructive pulmonary disease (COPD). The aim of the present study was to characterise the expression of FOXP3 in large and small airways of nonsmokers, smokers with normal lung function and COPD patients. A total of 19 nonsmokers, 20 smokers with normal lung function and 20 smokers with moderate COPD, undergoing lung resection for a solitary peripheral nonsmall cell carcinoma, were enrolled in the study. Immunohistochemical methods were used to evaluate FOXP3 expression in large and small airways. Smokers with normal lung function and COPD patients had increased numbers of FOXP3-positive cells in large airways compared with nonsmokers. A positive correlation was observed between FOXP3 expression in large airways and smoked pack-yrs. In small airways, COPD patients had decreased numbers of FOXP3-positive cells, compared with asymptomatic smokers and nonsmokers, that negatively correlated with airflow obstruction. To conclude, chronic obstructive pulmonary disease is characterised by upregulation of FOXP3-positive cells in large airways but a downregulation in small airways that correlated with airflow limitation. The results of the present study contribute to a better understanding of the pathogenesis of chronic obstructive pulmonary disease.

Patterns of inflammatory responses in large and small airways in smokers with and without chronic obstructive pulmonary disease.

Background: Chronic obstructive pulmonary disease (COPD) is characterised by progressive and irreversible airway obstruction. Smoking causes persistent inflammation in lung tissue. However, differences in inflammatory responses between the large and small airways have not been systematically explored among smokers with and without COPD. Objectives: The aim of our research was to characterise the expression and localisation of NF-ĸBp65 and histone deacetylase 2 (HDAC2) as well as inflammatory cell (macrophages, lymphocytes, neutrophils) distribution in large and small airways, in nonsmokers and in smokers with and without COPD. Methods: Nineteen nonsmokers, 20 smokers with normal lung ventilation function and 20 smokers with moderate COPD, undergoing lung resection for a solitary peripheral carcinoma, were enrolled in the study. Immunohistochemical methods were used to evaluate NF-ĸBp65 and HDAC2 expression and identify inflammatory cells in airways. Results: COPD patients had increased NF-ĸBp65 expression compared to nonsmokers and smokers without COPD, in both large and small airways, which corresponded to increased numbers of macrophages, CD8+ T lymphocytes and neutrophils. COPD patients had more macrophages in large compared to small airways and more CD8+ T lymphocytes and neutrophils in small compared to large airways. HDAC2 expression was significantly downregulated in smokers with COPD in small compared to large airways. Conclusions: Our findings indicate a nonuniform distribution of inflammatory cells throughout the bronchial tree. However, in both smokers with and without COPD, similar patterns of inflammatory processes occur in both large and small airways. The difference between smokers with and without COPD is only quantitative.

Breath testing as a method for detecting lung cancer

Early diagnosis of lung cancer is important due to high mortality in late stages of the disease. An ideal approach for population screening could be the breath analysis, due to its non-invasiveness, simplicity and cheapness. Using sensitive methods of analysis like gas chromatography/mass spectrometry in exhaled air of cancer patients were discovered some volatile organic compounds – possible candidates for cancer markers. However, these compounds were not specific for cancer cells. At the same time, integrative approaches used to analyze the exhaled breath have demonstrated high sensitivity and specificity of this method for lung cancer diagnosis. Such integrative approaches include detection of smell prints by electronic nose or integrated analysis of wide range of volatile organic compounds detected by gas chromatography/mass spectrometry or related methods. Modern statistical pattern recognition systems like logistic regression analysis, support vector machine or analysis by artificial neuronal network may improve diagnostic accuracy.

Distinct Influence of Atypical 1,4‐Dihydropyridine Compounds in Azidothymidine‐Induced Neuro‐ and Cardiotoxicity in Mice Ex Vivo

This study demonstrates the effective protection by compounds of atypical 1,4-dihydropyridine (DHP) series cerebrocrast, glutapyrone and tauropyrone against neuro- and cardiotoxicity caused by the model compound azidothymidine, a well-known mitochondria-compromising anti-HIV drug. In previous in vitro experiments, we have demonstrated distinct effects of these DHP compounds to influence mitochondrial functioning. In the present in vivo experiments, DHP compounds were administered intraperitoneally in mice daily for 2 weeks, per se and in combinations with azidothymidine at doses: azidothymidine 50 mg/kg; cerebrocrast 0.1 mg/kg; glutapyrone 1 mg/kg; and tauropyrone 1 mg/kg. At the end of the experiment, mice were killed, heart and brain tissues were removed and examined ex vivo histopathologically and immunohistochemically. NF-kappaBp65 and caspase-3 were used as the markers indicating inflammatory and apoptotic events, respectively. Cerebrocrast (dicyclic structure) was the most potent DHP, which effectively reduced azidothymidine-induced overexpression of NF-kappaBp65 and caspase-3 in mouse myocardium and brain cortex. Glutapyrone per se increased the number of caspase-3-positive cells in the brain, whereas it reduced NF-kappaBp65 and caspase-3 expression in cardiac tissue caused by azidothymidine. Tauropyrone showed dual action: per se it increased caspase-3 in the brain and NF-kappaBp65 expression in the heart, but it considerably reduced these activations in azidothymidine-treated mice. This study provides the first demonstration of a distinct pharmacological action for atypical DHP compounds in cardiac and brain tissues. The dicyclic structure of cerebrocrast is considered beneficial for neuro- and cardioprotection at least in part via mitochondrial targeting and consequent regulation of inflammatory and apoptotic processes.

Exhaled Air Analysis in Patients with Different Lung Diseases Using Artificial Odour Sensors

Exhaled Air Analysis in Patients with Different Lung Diseases Using Artificial Odour Sensors Sniffing breath to diagnose a disease has been practiced by doctors since ancient times. Nowadays, electronic noses are successfully used in the food, textile and perfume industry as well as for air pollution control. The aim of this study was to test whether exhaled breath analysed by an artificial nose could identify and discriminate between different lung diseases. A total of 76 individuals were tested: 25 bronchial asthma, 19 lung cancer, 10 pneumonia, 6 chronic obstructive pulmonary disease (COPD) patients and 16 healthy volunteers. Exhaled air was collected in plastic bags and immediately analysed using an electronic nose instrument (9185, Nordic Sensors AB) containing 14 different odour sensors. Multifactor logistic regression analysis was used to determine correlation between the amplitudes of sensor responses and the clinical diagnoses of patients and to calculate sensitivity and specificity of the method for each diagnosis. For diagnostics of asthma the sensitivity was found to be 84% and specificity — 86%. For lung cancer, the sensitivity was 74% and specificity, 95%; for pneumonia 90% and 98%, but for COPD, 33% and 97%, respectively. We conclude that an artificial nose is able to discriminate among different lung diseases with sufficiently good accuracy. This method may be further developed to implement it in clinical medicine for express diagnostics of acute and chronic lung diseases. Izelpas Gaisa Analīze Pacientiem Ar Dažzādām Plaušu Slimībām, Izmantojot Mākslīgos Ožas Sensorus Jau kopš seniem laikiem ārsti ostījuši pacienta elpu, lai uzstādītu slimības diagnozi. Mūsdienās ir radīts elektroniskais deguns, un to izmanto gan smaržu, gan tekstilrūpniecībā, gan lai novērtētu vides piesārņotību. Mūsu pētījuma mērķis bija noskaidrot, vai mākslīgo degunu var izmantot plaušu slimību diferenciāldiagnostikai. Pētījumā piedalījās 76 personas: 25 bronhiālās astmas slimnieki, 19 bija plaušu vēzis, 10 - pneimonija, 6 - hroniska obstruktīva plaušu slimība (HOPS), bet 16 bija veseli brīvprātīgie. Izelpas gaisu savāca plastikāta maisos un to tūlīt analizēja αr elektroniskā deguna aparātu (Nordic Sensors AB, 9185), kura sastāvā bija 14 smaržu sensori. Lai konstatētu korelāciju starp smaržas sensoru rādījumiem un konkrētām pacientu diagnozēm, kā arī lai noteiktu metodes specifiskumu un jutību, tika lietota multifaktoru logistiskās regresijas analīze. Jutība, paredzot astmas diagnozi, bija 84%, bet specifiskums - 86%. Jutība, nosakot plaušu vēzi, bija 74%, specifiskums - 95%; pneimonijai 90% un 98%, bet HOPS - attiecīgi 33% un 97%. Secinājām, ka elektroniskais deguns spēj atšķirt dažādas plaušu slimības ar pietiekamu precizitāti. Metodi varētu turpmāk attīstīt un lietot klīniskajā praksē akūtu un hronisku plaušu slimību ekspresdiagnostikai.

Detection of lung cancer in exhaled breath with an electronic nose using support vector machine analysis

Lung cancer is one of the most common malignancies and has a low 5-year survival rate. There are no cheap, simple and widely available screening methods for the early diagnostics of lung cancer. The aim of this study was to determine whether analysis of exhaled breath with an artificial olfactory sensor using support vector analysis can differentiate patients with lung cancer from healthy individuals and patients with other lung diseases, regardless of the stage of lung cancer and the most common comorbidities. Patients with histologically or cytologically verified lung cancer, healthy volunteers and patients with other lung diseases (e.g. chronic obstructive pulmonary disease (COPD), asthma, pneumonia, pulmonary embolism, benign lung tumors) were enrolled in the study. Breath sample collection and analysis with a Cyranose 320 sensor device was performed and data were further analyzed using a support vector machine (SVM). The SVM correctly differentiated between cancer patients and healthy volunteers in 98.8% of cases. The cancer versus non-cancer group patients (healthy volunteers and patients with other lung diseases) were classified correctly by SVM in 87.3% of cases. In the mixed diagnosis groups (only cancer, only COPD, cancer + COPD and control) all 79 out of 79 patients were predicted correctly in the cancer + COPD group, with the rate of correct prognosis in other patient groups being lower. Exhaled breath analysis by electronic nose using a SVM is able to discriminate patients with lung cancer from healthy subjects and mixed groups of patients with different lung diseases. It can also provide a certain level of discrimination between lung cancer patients, lung cancer patients with concomitant COPD, COPD alone and a healthy control group.

Increased innate and adaptive immune responses in induced sputum of young smokers

BACKGROUND AND OBJECTIVES It is known that chronic obstructive pulmonary disease (COPD) development process is imperceptible and can be asymptomatic for 20 or more years. It is of great importance to diagnose early inflammatory changes that can lead to COPD in young asymptomatic cigarette smokers. The aim of our study was to analyze the cell spectrum of induced sputum (IS) of young cigarette smokers, with emphasis on T-regulatory cells. MATERIALS AND METHODS A total of 20 healthy nonallergic smokers, 20 nonsmokers and 20 COPD patients were enrolled in the study. After lung function measurements were taken, we performed sputum induction and analyzed sputum cells. We evaluated the cell count of FOXP3-positive, CD4+ and CD8+ T lymphocytes by immunocytochemistry staining, and the cell count of macrophages and neutrophils by May-Grünwald Giemsa staining. RESULTS Induced sputum of smokers contained a higher absolute amount of macrophages and neutrophils when compared to nonsmokers. FOXP3-positive cells in the sputum of young smokers showed a statistically significant increase when compared to nonsmokers. Induced sputum of COPD patients contained an increased absolute amount of neutrophils and FOXP3-positive Treg cells when compared to nonsmokers. Regression analysis showed that the amount of FOXP-3 positive cells, neutrophils and macrophages in the induced sputum was increasing with the number of pack years. CONCLUSIONS This study demonstrates that young smokers have early inflammatory changes in their airways that not only initiate nonspecific mechanisms recruiting neutrophils, but also involve specific immune mechanisms with recruitment of T regulatory lymphocytes. The lymphocyte response is probably adaptive.

35 ROLE OF CXCL4 CHEMOKINE IN ANGIOGENIC ACTIVITY OF NON-SMALL CELL LUNG CANCER (NSCLC)

35 ROLE OF CXCL4 CHEMOKINE IN ANGIOGENIC ACTIVITY OF NON-SMALL CELL LUNG CANCER (NSCLC) A. Spaks1, D. Isajeva2, I. Spaka3, A. Pirtnieks1, I. Jaunalksne4, J. Grusina-Ujumaza1, J. Basko1, I. Taivans2, D. Krievins5 1Thoracic Surgery, Pauls Stradins Clinical University Hospital, Riga, Latvia, 2Pathology, University of Latvia, Riga, Latvia, 3Biology, Riga Stradins University, Riga, Latvia, 4Clinical Immunology, Pauls Stradins Clinical University Hospital, Riga, Latvia, 5Surgery, University of Latvia, Riga, Latvia

H1-antihistamines suppress wheal-and-flare reaction and skin blood perfusion measured by Laser Dopppler flowmetry: randomized, double-blind, placebo-controlled, crossover design study

H1-antihistamines suppress wheal-and-flare reaction and skin blood perfusion measured by Laser Dopppler flowmetry: randomized, double-blind, placebo-controlled, crossover design study The aim of our study was to compare the influence of pre-treatment with H1-antihistamines (levocetirizine, desloratadine, clemastine, quifenadine, and sequifenadine) and a placebo on the histamine-induced weal and flare reaction, increase of skin blood perfusion and sedation. Thirty healthy volunteers were enrolled in the study. The study design was a prospective, randomised, double-blind, placebo-controlled, crossover, balanced clinical trial. Volunteers in randomised and double-blind order were treated with oral levocetirizine 5 mg, desloratadine 5 mg, clemastine 1 mg, quifenadine 50 mg, sequifenadine 50 mg or a placebo. Two hours after intake of medication, the histamine skin prick test was performed and skin blood perfusion was registered with further evaluation of sedative effect. We conclude that levocetirizine induced a significant and pronounced decrease of weal and flare reaction and skin blood perfusion compared to the placebo and the other H1-antihistamines. The effect of quifenadine and sequifenadine on weal reaction area was similar to desloratadine and clemastine. Regarding the sedative effect, we can conclude that second generation antihistamines appear to be not non-sedative but the least impairing, and the first generation antihistamines appear to be the most impairing on central nervous system function. There is a necessity to consider the sedating potential of antihistamines, along with other factors such as efficacy, when prescribing antihistamines to patients. H1-antihistamīni samazina papulas un eritēmas reakciju un asins perfūzijas pieaugumu ādā, izvērtējot ar lāzera Doplera plūsmas analīzes metodi: randomizēts, dubultakls, placebo kontrolēts krustota dizaina pētījums Mūsu pētījuma mērķis bija salīdzināt ietekmi, kāda piemīt pirms testa lietotiem H1-antihistamīniem — levocetirizīnam, desloratadīnam, klemastīnam, kvifenadīnam, sekvifenadīnam — un placebo uz histamīna izraisītu papulas un eritēmas reakciju, asins perfūzijas pieaugumu ādā un sedāciju. Pētījumā tika iesaistīti 30 veseli brīvprātīgie. Šis bija prospektīvs, randomizēts, dubultakls, placebo kontrolēts, krustota dizaina un balansēts klīnisks pētījums. Brīvprātīgajiem randomizētā un dubultaklā veidā tika doti perorāli 5 mg levocetirizīna, 5 mg desloratadīna, 1 mg klemastīna, 50 mg kvifenadīna, 50 mg sekvifenadīna vai placebo. Divas stundas pēc kapsulas lietošanas tika veikts ādas dūriena tests ar histamīnu, tika registrētas asins perfūzijas izmaiņas ādā, kā arī sedatīvā efekta izvērtēšana. Mēs secinājām, ka levocetirizīns izraisa visnozīmīgāko un visizteiktāko ādas papulas un eritēmas reakcijas samazinājumu, kā arī visievērojamāko asins perfūzijas samazinājumu ādā, salīdzinot ar placebo un citiem H1-antihistamīniem. Kvifenadīna un sekvifenadīna ietekme uz papulas reakciju bija līdzīga desloratadīna un klemastīna ietekmei. Attiecībā uz sedatīvo efektu var secināt, ka otrās paaudzes antihistamīni ir nevis bez sedatīvas ietekmes, bet to sedatīvā ietekme atstāj vismazāko iespaidu un sekas uz organisma funkcijām, savukārt pirmās paaudzes antihistamīni visvairāk pasliktina centrālās nervu sistēmas funkciju. Jāievēro, ka, izrakstot pacientam kādu H1-antihistamīna preparātu, jāatceras arī par to sedatīvo ietekmi, ne tikai tiešo efektu uz alergiskās reakcijas patogenētiskajiem faktoriem.

REGIONAL LUNG BLOOD PERFUSION MEASURED WITH LASER DOPPLER METHOD DURING BODY POSITION CHANGE, VALSALVA MANOEUVRE AND CARDIOPULMONARY BYPASS

Regional Lung Blood Perfusion Measured With Laser Doppler Method During Body Position Change, Valsalva Manoeuvre and Cardiopulmonary Bypass Methodological approaches to investigate local regulatory mechanisms of lung blood supply in humans are restricted. We tried a new approach using laser Doppler technique. During bronchoscopy, an angled laser Doppler flow probe was introduced through a biopsy channel and wedged into small bronchus. Laser light penetrated the wall of small bronchus and was reflected from blood cells running through neighbouring capillaries. Regional blood perfusion changes were recorded during body position change from vertical to supine and back to vertical, while performing Valsalva maneuver and during cardiac bypass surgery. Body position change and Valsalva maneuver markedly influenced the blood perfusion signal. During cardiac bypass when lungs were supplied with blood only through bronchial arteries regional blood perfusion dropped substantially on average from 93 ± 42 to 7.3 ± 4.3 perfusion units. We conclude that blood perfusion measured with this method reflect mainly the pulmonary vascular bed and may be used for investigation of its local regulatory mechanisms. Regionālās Plaušu Perfūzijas Pētījums AR Lāzera Doplera Metodi, Ķermenim Atrodoties Daādās Pozās, Veicot Valsalva Manevru UN Mākslīgās Asinsrites Laikā Plaušu asinsrites regulācijas mehānismi cilvēkam līdz šim maz pētīti, jo trūkst atbilstošu metožu. Pētījumā izmantota origināla pieeja, kas ļāva registrēt plaušu regionālo asinsriti, ieķīlējot lāzera Doplera leñķa zondi vienā no sīkajiem bronhiem bronhoskopijas laikā. Mērījumi tika veikti, cilvēkam atrodoties dažādās ķermeña pozās, kā arī veicot Valsalva manevru un sirds operācijās mākslīgās asinsrites laikā. Ķermeña pozu maiña un Valsalva manevrs būtiski ietekmē asins perfūziju plaušās. Mākslīgās asinsrites laikā plaušu asinsriti uztur tikai regionālā perfūzija bronhiālo artēriju līmenī, un tad perfūzija mazinās no 93 ± 42 līdz 7,3 ± 4,3 perfūzijas vienībām. Secināts, ka, izmantojot šādu metodiku, galvenokārt iegūst signālu no mazā asinsrites loka. Šādu metodiku var izmantot, lai pētītu lokālos asinsrites regulācijas mehānismus plaušās.