Comparative Expression of Apoptotic Markers in Lung Adenocarcinoma and Squamous Cell Carcinoma.

Lung cancer is still an oncology challenge. A 5-year survival reaches less than 20 % of patients. Apoptosis disturbances are a key step in cancer development. The evaluation of apoptosis markers has a great potential in lung cancer. The goal of our study was a comparative evaluation of apoptosis regulators: p53, Bcl-2, Bax, COX-2, and survivin in lung adenocarcinoma (AC) and squamous cell carcinoma (SCC). We also evaluated the relationship between apoptosis markers and clinicopathological parameters. Fifty six patients with non-small cell lung cancer (NSCLC) were included into the study (20 women and 36 men). AC was diagnosed in 30 and SCC in 26 cases. The evaluation of markers was performed using an immunohistochemical method on paraffin embedded tissue specimens. We used monoclonal antibodies for p53, bcl-2, and COX2-proteins (clone DO7, bcl-2/100/D5, and 4H12, respectively), Bax (B-9 clone) and survivin (clone 12C4). The results of immunostaining were viewed by light microscopy. We revealed significantly more frequent expression of Bax and survivin in lung AC than SCC (p < 0.01 and p < 0.019). Bcl-2 immunoreactivity was seen more often in AC without lymph node metastases than with metastases (p = 0.046). There was no correlation between the apoptosis markers and gender or the presence of vessel emboli. A greater variability in markers expression was seen in lung AC than SCC. There were significant differences in the Bax and survivin expression in the two major pathological types of NSCLC. We did not revealed any correlation between the markers and TNM characteristics, accept for Bcl-2 presence along with the lymph node involvement in the AC group.

Correlations between osteoprotegerin serum levels and body composition parameters in patients with sleep apnea syndrome and the possible influence on cardiovascular risk.

INTRODUCTION: Osteoprotegerin (OPG) is a member of the tumor necrosis factor family and a key regulator in bone turnover; it plays a role in the development of many cardiovascular diseases and may be treated as a marker of vascular damage. Bioelectrical impedance analysis (BIA) is a reliable, non-invasive and effective technique for measuring body composition. The aim of the study was to evaluate correlations between osteoprotegerin serum levels and body composition parameters in sleep apnea patients and their influence on cardiovascular risk. MATERIAL AND METHODS: A total of 125 patients with newly diagnosed OSA were enrolled in the study (including 34 females). The mean age was 54.48±8.81 years, mean AHI 33.16±20.44/h and mean BMI 33.76±7.18. A control group comprised 59 healthy subjects with mean age of 51.27±12.97 years and mean BMI 29.47±5.42. All subjects underwent a nocturnal respiratory polygraphy and body composition measurements were taken with bioelectrical impedance analysis. OPG serum levels were measured using the enzyme-linked immunosorbent assay (ELISA) method. RESULTS: In OSA patients OPG correlated negatively with muscle mass percentage (MM%), phase angle, fat free mass percentage (FFM%) and body cell mass percentage (BCM%), while there was a positive correlation between osteoprotegerin and fat mass percentage (FM%). We demonstrated higher OPG serum levels in OSA patients with cardiovascular diseases than in those without comorbidities (4.01 vs 3.46pmol/l, p<0.05). CONCLUSION: Our findings, combined with previous observations in other diseases, suggest that elevated OPG serum levels together with selected body composition parameters may be helpful in identifying OSA patients with increased cardiovascular risk.

Genetic polymorphisms and expression of HLA-G and its receptors, KIR2DL4 and LILRB1, in non-small cell lung cancer.

Human leukocyte antigen-G (HLA-G) is a nonclassical HLA class I molecule absent from most normal tissues but detected in many malignant tumors. It is recognized by cells of the immune system using LILRB1, KIR2DL4 and LILRB2 receptors. We attempted to find out whether some polymorphisms of HLA-G, LILRB1 and KIR2DL4 genes are associated with susceptibility to nonsmall cell lung cancer (NSCLC). Four polymorphisms in HLA-G, i.e. -964A>G (rs1632947), -725C>G>T (rs1233334), -716T>G (rs2249863) in the promoter, and a 14 base pair insertion/deletion (14 bp indel) in the 3'-untranslated region (3'UTR), and five in LILRB1 - 5651G>A (rs41308748) in intron 14, 5717C>T L622L (rs1061684), 5724G>A E625K (rs16985478), 5774 C>A P641P (rs41548213) in exon 15, and 5806C>T (rs8101240) in 3'UTR - as well as 9620 9A/10A (rs11410751) polymorphism in exon 7 of KIR2DL4 were typed using different laboratory techniques. Only one single nucleotide polymorphism (SNP) in HLA-G (-964A>G) and one in LILRB1 (5724G>A) were found to influence the risk of NSCLC. In addition, 5724G>A was associated with protection from tumor cell infiltration of regional lymph nodes. Most importantly, we detected HLA-G and LILRB1 expression in tumor specimens, but no correlation with genetic polymorphisms was observed. HLA-G and LILRB1 protein expression levels in tumor tissue were significantly correlated with tumor stage.

Nocturnal parasympathetic modulation of heart rate in obesity-hypoventilation patients.

Heart rate variation (HRV) reflects the activity of the autonomic nervous system. The aim of the study was to analyze HRV in obstructive sleep apnea (OSA) and obesity hypoventilation (OH) patients to answer the question of whether chronic alveolar hypoventilation influences autonomic heart rate regulation. In 41 patients, diagnosed with either 'pure' OSA (n=23, apnea/hypopnea index--AHI: 43.8±18.0 PaCO2≤45 mmHg) or OH syndrome (n=18, AHI 58.7±38.0 PaCO2>46 mmHg), the HRV was analyzed, based on an 8 h ECG recording during sleep. In the OH patients, compared with the OSA patients, there was a globally decreased HRV, with a corresponding decrease in high frequency power in the spectral analysis of HRV and increased low frequency/high frequency ratio (p<0.05), indicating a reduced parasympathetic and increased sympathetic heart rate modulation. We conclude that hypoxemia and hypercapnia of the sleep disordered breathing have an impact on the autonomic heart rate regulation. HRV indices have a potential to become prognostic factors for the development of cardiovascular complications in patients with sleep disordered breathing.

Soluble ligand CD40 and uric acid as markers of atheromatosis in patients with obstructive sleep apnea.

The aim of the study was to evaluate the risk of atheromatosis in patients with obstructive sleep apnea (OSA), as based on the concentration of the pro-atherogenic soluble CD40L ligand (sCD40L) in relation to the level of uric acid. The serum levels of sCD40L and uric acid were measured in 79 OSA patients (mean apnea/hypopnea index - AHI 34.4 ± 20.9) and in 40 healthy controls. We found that sCD40L was higher in the OSA patients with hyperuricemia than that in the patients with uric acid in the normal range (sCD40L: 9.0 ng/ml vs. 8.0 ng/ml, respectively, p < 0.05). There was a positive association of sCD40L with AHI (p = 0.01) and a negative one with the mean minimal nocturnal SaO2(p < 0.05). Uric acid correlated negatively with the mean and minimal SaO2and positively with the oxygen desaturation index (p < 0.05). OSA patients with hyperuricemia also had a higher prevalence of hypertension and ischemic heart disease. We conclude that OSA patients with increased uric acid concentration have increased risk of atheromatosis, as indicated by a higher level of soluble pro-atherogenic ligand CD40, and a higher prevalence of cardiovascular adverse events.

Efficacy of noninvasive volume targeted ventilation in patients with chronic respiratory failure due to kyphoscoliosis.

Severe kyphoscoliosis can cause chronic respiratory failure. Noninvasive mechanical ventilation (NIMV) is a new optional treatment for such patients. The aim of this study was to evaluate the effectiveness of average volume-assured pressure support (AVAPS) NIMV in patients with kyphoscoliotic chronic respiratory failure. The study was performed in 12 patients (mean age 49±11 years and body mass index 27.5±7.9 kg/m2) with advanced kyphoscoliosis complicated by severe respiratory failure (PaO2 6.68±0.34 kPa, SaO2 81.7±3.1%, PaCO2 9.51±1.08 kPa) treated by the NIMV. The short-term, after 5 days, and long-term, after 1 year of home treatment, efficacy of NIMV was evaluated. We found a significant improvement of diurnal PaO2 and PaCO2 on the 5th day of NIMV (an increase of 1.4±0.3 kPa and a decrease of 1.8±0.8 kPa, respectively; p<0.05) and after one year NIMV (an increase of 2.07±0.46 kPa and a decrease of 2.68±0.85 kPa, respectively; p<0.05). There was a significant increase of mean blood oxygen saturation during sleep on the 5th day (86.2±3.2%) and after 1 year of treatment (89.4±2.1%) compared with the baseline level (83.2±3.2%). The forced vital capacity also increased after 1 year (1,024±258 ml vs. the baseline 908±267 ml; p<0.05). The NIMV was well tolerated and no patient discontinued the treatment during the observation period. We conclude that AVAPS NIMV is an effective treatment option in kyphoscoliotic patients with chronic respiratory failure, resulting in a prompt and long-term improvement of daytime and nocturnal blood gas exchange.

Efficacy of noninvasive mechanical ventilation in obese patients with chronic respiratory failure.

Chronic respiratory failure (CRF) develops in a minority of obese patients. Noninvasive mechanical ventilation (NIMV) is a new optional treatment for such patients. The aim of this study was to evaluate the effectiveness of NIMV in obese patients with CRF. The material of the study consisted of 34 obese patients (body mass index 47.3 ± 7.9 kg/m(2)) with CRF (PaO2 = 6.40 ± 0.93 kPa and PaCO2 = 8.67 ± 2.13 kPa) who were hypoxemic despite an optimal therapy. Thirteen patients had an overlap syndrome (OS) - chronic obstructive pulmonary disease (COPD) coexisting with obstructive sleep apnea syndrome (OSAS) and 21 patients had obesity-hypoventilation syndrome (OHS). Ventilation parameters were determined during polysomnography. The efficacy of NIMV was evaluated on the fifth day and after 1 year's home treatment. We observed a significant increase in the mean blood oxygen saturation during sleep in all patients; the increase was greater in patients with OHS (92.6 ± 1.4 %) than in patients with OS (90.4 ± 1.8 %). There was a significant improvement of diurnal PaO2 and PaCO2 on the fifth day of NIMV (mean PaO2 increase 2.1 kPa and PaCO2 decrease 0.9 kPa) and also after 1 year of home NIMV (mean PaO2 increase 1.9 kPa and PaCO2 decrease 2.4 kPa). Only one patient stopped treatment because of lack of tolerance during the observation period (1-3 years). In conclusion, NIMV is an effective and well tolerated treatment option in obese patients with CRF resulting in a rapid relief of respiratory disorders during sleep and a gradual, long-term improvement of gas exchange during the day, particularly in patients with OHS.

Serum antibodies to retinal antigens in lung cancer and sarcoidosis.

OBJECTIVE: Autoantibodies to various neuronal proteins frequently accompany lung cancer and their appearance may precede cancer symptoms. In this study we examined which retinal antigens (RAs) are recognized by sera of patients with lung cancer and whether the occurrence of serum antibodies to particular RAs is characteristic for cancer in comparison with a noncancer lung disease. METHODS: Sera of 72 patients with non-small-cell lung cancer (NSCLC), 29 with small-cell lung cancer (SCLC), 27 with sarcoidosis (S), and sera of 32 healthy donors were examined in immunoblotting using retinal extracts and purified RAs as antigens. RESULTS: 69.0% of SCLC, 45.8% of NSCLC, and 44.4% of S sera displayed anti-RAs reactivity. Significantly less (p < 0.05; chi(2) test) percent of healthy control sera reacted with RAs. Lung cancer sera recognized mainly 46-, 56-, and 36-kD and to a smaller extent also 96-, 72-, 43-, and 26-kD proteins. Most of them were recognized with about 2-fold lower frequencies by S and control sera. Only lung cancer sera contained very high-titer antibodies to 46- and 26-kD RAs, identified as alpha-enolase and recoverin, respectively. CONCLUSION: Antibodies to RAs occur more frequently and in higher titers in lung cancer (especially SCLC) than in sarcoidosis or control sera. Although antibodies to retinal alpha-enolase, recoverin and other RAs are present mainly or exclusively in lung cancer sera, none of them seems to be a specific marker of a particular disease.

Serum thrombopoietin levels in patients with reactive thrombocytosis due to lung cancer and in patients with essential thrombocythemia.

In patients with thrombocytosis normal to increased serum thrombopoietin (TPO) levels have been reported. The aim of this study was to investigate the relationship between serum TPO concentration, platelet number and plasma levels of fibrinogen in patients with reactive thrombocytosis (RT) due to lung cancer and in patients with essential (primary) thrombocythemia (ET). A total of 70 newly diagnosed patients with RT or ET (platelet counts >600 G/l) were studied: 45 with RT due to lung cancer (25 with non-small cell lung cancer, NSCLC and 20 with small cell lung cancer, SCLC), and 25 with ET. Twenty normal volunteers were used as controls. TPO was measured by immunoassay technique (ELISA). Mean serum TPO values in patients with RT due to lung cancer were statistically significantly higher than those in patients with ET or in controls. The highest platelet count was seen in patients with ET, and mean plasma fibrinogen levels were the highest in RT patients. In NSCLC patients mean serum TPO concentrations were higher (not statistically significant) than in SCLC, and a statistically significant relationship between TPO serum concentration and fibrinogen level was observed. No correlations between platelet counts and TPO serum concentrations were found. Our results indicate increased serum TPO levels in patients with thrombocytosis in lung cancer which may be related to the activity of neoplasms. In addition, it is postulated that the relatively low TPO values in patients with ET may result from a dysregulation of the feedback loop involved in platelet production.

[Pulmonary alveolar microlithiasis in a patient with urolithiasis and cholelithiasis]. / Kamica pecherzyków plucnych u chorej z kamica ukladu moczowego i zólciowego.

49 years old woman was admitted to Pulmonary Ward because of respiratory and cardiac failure. Six years earlier she was surgically treated because of urolithiasis and at this time disseminated lesions in both lungs were revealed. Biopsy made during transbronchial procedure showed microlithiasis. She had'nt any respiratory symptoms during 6 years. Now she had PaO2 = 36 mmHg and massive infiltrations in whole lungs. Palliative therapy with diuresis, cardiac drugs and oxygen therapy diminished symptoms of respiratory and cardiac failure.

[Atrial natriuretic excretion in patients with obstructive sleep apnea syndrome (OSAS)]. / Wydzielanie hormonu natriuretycznego w zespole obturacyjnego bezdechu sródsennego.

UNLABELLED: During obstructive sleep apneas stimuli, that may increase excretion of atrial natriuretic peptide (ANP) occur. The aim of the study was the evaluation whether in patients with OSAS levels of ANP are significantly different in relation to sleep or wakefulness and in relation to disturbances of ventilation during sleep and wakefulness. The material of the study consisted of 34 patients with OSAS (age 25-65 years). There were no differences in the levels of ANP late in the evening, during sleep and early in the morning. There were 2 groups of the patients: with low (< 70 pg/ml, mean at 21 p.m. 9.7 +/- 8.7 pg/ml, at. 2 a.m. 12.5 +/- 9.3 pg/ml, at 6 a.m. 14.4 +/- 15.1 pg/ml) and high (> 70 pg/ml, mean at 21 p.m. 148.6 +/- 232.9 pg/ml, at 2 a.m. 119.5 +/- 45.5 pg/ml, at 6 a.m. 164.9 +/- 161 pg/ml) ANP levels. As compared with patients with low ANP levels, patients with high ANP levels were older and more obese, more frequently had concomitant COPD, lower VC and FEV1, higher daytime PaCO2 and lower PaO2; most of them had peripheral edema. In patients with high ANP levels there was more profound mean arterial blood desaturation during sleep apnoeas than in patients with low ANP levels (SaO2 75 +/- 8% vs 81 +/- 4%, p < 0.001), although apnea index and mean apnea duration were similar in both groups. CONCLUSIONS: In patients with OSAS the daytime and sleep levels of ANP are similar. High levels of ANP can be found in OSAS patients with impaired daytime ventilation and gas exchange, and profound arterial oxygen desaturation during sleep apnoeas.

[Sarcoidosis with central nervous system involvement]. / Sarkoidoza z zajeciem osrodkowego ukladu nerwowego.

A case of pulmonary sarcoidosis with central nervous system involvement was described. Diagnosis was based on hilar lymph nodes biopsy and OUN MRI scan. Patient was treated with glicocorticosteroids with improvement.

[Breast tuberculosis]. / Gruzlica sutka.

A 47-year-old woman, mother of 7 children, was admitted to hospital because of the painful tumour of her left breast. The clinical course was chronic with formation of abscess and then fistula. Chest X-ray was normal. Axillary lymph nodes were not palpable. During mammography tumour was revealed (3.5 cm in diameter). During tumour resection abscess was found. Staphylococcus aureus was cultured from its pus, but histologic picture of resected tissue suggested caseous tuberculosis of mamma. Bacilli were not found. The patient was treated with antituberculous drugs. At the early stage of treatment the skin fistula of mamma occurred but the long-lasting therapy was successful.

Histamine receptor expression on peripheral blood lymphocytes is influenced by specific and nonspecific activation.

Histamine is a physiological mediator which exerts both effector and regulatory functions through its receptors on various cells. The aim of the study was to investigate changes in histamine receptor expression on peripheral blood lymphocytes affected by stimulation with both specific and nonspecific stimuli. Lymphocytes were obtained from both healthy and allergic subjects. Cells were incubated with various allergens (mixed grass pollen, Lolium perenne, Dermatophagoides pteronyssinus 1, bee venom, phospholipase A2) and nonspecific (fMLP, PMA/ionomycin, LPS) stimuli. The percentage of histamine-binding cells was determined with a fluorescence microscope after incubation with histamine-fluorescein. In control subjects histamine binding after stimulation with allergens was not significantly changed. In contrast, in allergic subjects stimulation with specific allergens resulted in significantly increased histamine binding. Nonspecific stimulation caused increased histamine binding to lymphocytes in both allergic subjects and healthy controls. We conclude that specific and nonspecific activation of lymphocytes is associated with increased expression of histamine receptors.

The contribution of endotoxins present in the respiratory tract to overproduction of nitric oxide associated with impaired interleukin 6 release in bronchoalveolar leukocytes from lung cancer patients.

The purpose of the study was to assess the relation between the levels of endotoxins circulating in airways of patients with lung cancer and the ability of bronchoalveolar lavage (BAL) leukocytes for ex vivo release of nitric oxide (NO) and interleukin 6 (IL-6) and for in vitro lipopolysaccharide (LPS)-induced production of the mediators. Leukocytes isolated from the BAL of 11 patients and from 5 healthy individuals were cultured in the absence or presence of LPS(E. coli). The levels of endotoxins in the BAL fluids (BALF) and the amounts of NO released ex vivo from unstimulated cells from the patients were highly (p = 0.0025) elevated in comparison with those from healthy individuals. The release of NO was significantly correlated (R(S) = 0.638, p = 0.047) with the levels of endotoxins in BALF. In contrast, production of IL-6 remained very low and a negative correlation (R(S) = -0.623, p = 0.0542) was observed between the amounts of NO and IL-6. It was also found that, in response to LPS, bronchoalveolar leukocytes from patients with lung cancer express a reduced capacity for in vitro production of NO and IL-6. Our data suggest that, in patients with lung cancer, the activation of BAL cells by endotoxins circulating in the airways may contribute, at least in part, to overproduction of spontaneous NO and, subsequently, the NO may reduce IL-6 production. Moreover, the exposure in vivo of the BAL cells to LPS renders them unable to respond to the second signals.

Heparin modulates migration of human peripheral blood mononuclear cells and neutrophils.

Evidence has now accumulated that heparin can significantly affect immune response including allergic inflammation. Cell motility is supposed to be very crucial in this process. Thus the aim of our study was to investigate whether heparin is a chemoattractant for some inflammatory cells and is also capable of influencing chemotaxis induced by typical chemoattractants. Peripheral blood mononuclear cells (PBMCs) and neutrophils from 10 healthy subjects were obtained by gradient centrifugation. Chemotaxis assays towards either heparin (molecular weight 16 kDa) or low molecular weight heparin--fraxiparine (molecular weight 5 kDa) were performed in Boyden chambers. We found that both heparin molecules are chemoattractants for both PBMCs and neutrophils in the wide concentration range (0.1-2000 microg/ml). However, maximal chemotaxis was observed at concentrations 50-100 microg/ml (fraxiparine) and 1-50 microg/ml (heparin). We also found that fraxiparine was able to significantly increase chemokinesis and decrease chemotaxis in the gradient of both fMLP and IL-8. These results indicate that heparin is a potent regulator of cell migration.

[Endothelin concentration in sera of patients with atopic bronchial asthma]. / Stezenia endoteliny w surowicy chorych na astme oskrzelowa atopowa.

Endothelin-1 (ET-1) is an effective vasoconstrictor and has bronchoconstricting property. Recent findings in vivo and in vitro indicate the influence of ET-1 on bronchial smooth muscle tone. ET-1 concentration was detected in BAL-Fluid in patients with bronchial asthma. Previous studies indicated an increase in ET-1 serum concentrations in the course of exacerbation of asthma. The purpose of our study was to compare ET-1 concentrations in sera of asthmatics during the asymptomatic period and after metacholine provocation. The study was performed in a group of 16 patients with mild bronchial asthma and in 11 healthy subjects. ET-1 concentrations in sera were evaluated by Elisa method (kit R&D USA). Bronchial provocation with metacholine was performed in asthmatics using Bronchoscreen. The result was considered as positive with FEV1, decrease of at least 20%. Airway responsiveness to metacholine was expressed as PC20. The results were analyzed statistically by means of the Student's test. The baseline ET-1 levels in sera of healthy donors were lower then in asthmatics group (x1' = 4.72 +/- 1.01, x1 = 11.53 +/- 3.68 pg/ml, p < 0.001). We found a statistically significant increase of ET1-1 concentration in sera after inhalation of metacholine (x1 = 11.55 +/- 3.68; x2 = 24.08 +/- 4.09 pg/ml, p < 0.001). The results indicate that ET-1 may play a role in bronchospasm in asthmatics.