[Higher incidence of alveolar lymphocytes (AL) apoptosis in smokers depends on BCL-2 expression and specific response to tumor necrosis factor alpha (TNFalpha). Bronchoalveolar lavage (BAL) material analysis from selected interstitial lung diseases (ILD) and healthy controls]. / Czestsza apoptoza limfocytów pecherzykowych (alveolar lymphocytes, AL) u palaczy papierosów zalezy od ekspresji BCL-2 i swoistej odpowiedzi na czynnik martwicy nowotworów alpha (tumor necrosis factor alpha, TNFalpha). Analiza materialu z plukania oskrzelowo-pecherzykowego (bronchoalveolar lavage, BAL) chorych z wybranymi chorobami sródmiazszowymi pluc i osób zdrowych.

BACKGROUND: We have previously described the increased apoptosis rate in smokers alveolar lymphocytes (AL) that was independent from the FASL/ FAS system activation. Consequently, the role of intrinsic apoptosis pathway and other ligand/death receptor pairs as TNFalpha/TNFR1 and TRAIL/DR4 important for apoptosis regulation should be considered in this phenomenon. The purpose of the study was to evaluate the impact of tobacco consumption on expression of selected BCL-2 family members and ligand/receptors pairs in bronchoalveolar lavage (BAL) harvested from patients with pulmonary sarcoidosis (PS), idiopathic pulmonary fibrosis (IPF) and healthy volunteers. The results were analyzed in the context of AL apoptosis rate. METHODS: AL apoptosis from PS (n=36, incl. 22 smokers), IPF (11, incl. 5 smokers) and controls (n=17, incl. 9 smokers) was evaluated by flow cytometry (sub-G1 of cell cycle). AL were stained for BCL-2, BCL-xL, BAK, TNFR1 (CD120A) TNFR2 (CD120B) and DR4. ELISA assay was used to evaluate the BAL supernatant levels of TNFalpha and TRAIL. RESULTS: According to previous observations, AL apoptosis rate was significantly higher in smoker subgroups as compared to nonsmoking counterparts. Decreased AL BCI-2+ relative number was observed in smoking PS (80.5 +/- 6.2 vs 91 +/- 9.8% in nonsmokers) and controls (59 +/- 14.1% vs 75 +/- 16.1%, p<0.05). TNFalpha concentration in BAL supernatant was significantly higher only in healthy smokers (2.32 +/- 0.77 vs 0.42 +/- 0.27 pg/ml, p<0.05), whereas TRAIL levels were remarkably enhanced in IPF smokers (44.8 +/- 12.8 vs 13.5 +/- 5.0 pg/ml, p<0.05) only. However, TUNEL. detected AL apoptosis positively correlated with TNFalpha. in smokers (p<0.05) and negatively with AL CD120B:CD120A expression ratio. Paradoxically, TNFalpha levels were positively correlated with AL BCL-2 expression in nonsmokers (Rs +0.58, p<0.01), but not in smokers. No differences were observed in all subgroups in respect to AL expression of DR4, BCL-xL or BAK. CONCLUSIONS: 1. AL were not sufficiently protected against apoptosis in smokers. 2. The most likely mechanisms involve down-regulation of BCL-2 expression and altered AL susceptibility to TNFalpha, mediated by imbalance between AL membrane expression of TNF receptor type 1 (death receptor) and type 2 (survival mediator). 3. Mechanisms regulating the increased AL apoptosis in smokers seem to be different in each tested group.

[Acute clozapine poisonings in years 2007 - 2010 in material of Clinic of Toxicology in Kraków]. / Ostre zatrucia klozapina w materiale Kliniki Toksykologii w Krakowie z lat 2007 - 2010.

18 patients with acute clozapine poisoning, 6 female and 12 male, were analyzed. The mean age was 42.8 years. Six patients were intoxicated only clozapine. Mixed poisoning (clozapine and other factor) was diagnosed in nine cases. Among the additional factors dominated psychotropic drugs. According to the Poisoning Severity Score (PSS) criteria in the study group was only a one mild intoxication. Acute pneumonia developed in 3 patients, acute bronchitis and rabdomyolysis were reported in one case. The most common symptoms included: agitation, confusion (83.3%), tachycardia (77.8%), CNS depression (66.7%), excessive mucus production in bronchi, hypersalivation (44.4%), miosis (50%). Disordered breathing requiring intubation or mechanical ventilation occurred in 27.7% of poisoned. The average duration of hospitalization was less than 7 days.

[Cardiologic interaction carbamazepine with atypical neuroleptics. Acute mixed carbamazepine and quetiapine intoxication--case report]. / Kardiologiczne interakcje karbamazepiny z atypowymi lekami przeciwpsychotycznymi. Opis przypadku ostrego zatrucia mieszanego karbamazepina i kwetiapina.

The treatment of bipolar disorders is based on the concomitant use of the mood stabilizers and antipsychotics. In the first case carbamazepine is frequently used, in the second case second-generation (atypical) neuroleptics like quetiapine. This drug combination in a significant number of cases prevent recurrence of mania and depression, normalizes mood and emotion and improves lifestyle. However, this treatment is related to increased interactions and risk of the side effects. Even greater risks involve mixed drug intoxications with these groups of drugs. In this paper we present acute poisoning with carbamazepine and quetiapine complicated cardiotoxic effects in the form of arrhythmias and conduction disorders of the heart. This symptoms disappeared spontaneously after resolution of the poisoning.

Enhanced expression of Fas Ligand (FasL) in the lower airways of patients with fibrotic interstitial lung diseases (ILDs).

The exact role of FasL, and particularly its soluble and membrane-bound forms, in the development of chronic ILDs and lung fibrosis has not been extensively explored. We aimed at analyzing membrane-bound FasL expression on alveolar macrophages (AM) and lymphocytes (AL) as well as soluble FasL (sFasL) levels in bronchoalveolar lavage (BAL) from ILDs patients, incl. pulmonary sarcoidosis (PS), hypersensitivity pneumonitis (HP), silicosis, asbestosis, idiopathic pulmonary fibrosis (IPF), nonspecific interstitial pneumonia (NSIP), and healthy subjects (n = 89, 12, 7, 8, 23, 6, 17, respectively). In IPF, significantly increased percentage of AM FasL(+) and CD8(+)FasL(+) cells as well as sFasL levels in BAL were found. Increased sFasL levels were also observed in HP. NSIP and asbestosis were characterized by higher AM FasL(+) relative number; CD8(+)FasL(+) population was expanded in asbestosis only. There was a significant decline in AL FasL(+) percentage in PS and HP. Vital capacity was negatively correlated with sFasL levels, AM FasL(+) and CD8(+)FasL(+) cell relative count. CD4(+)FasL(+) and CD8(+)FasL(+) percentage strongly correlated with BAL neutrophilia, an unfavorable prognostic factor in lung fibrosis. The concurrent comparative BAL analysis of FasL expression indicates that FasL(+) AM and AL (mainly Tc cells) comprise an important element of the fibrotic process, mostly in IPF. FasL might play a crucial role in other fibrosis-complicated ILDs, like NSIP and asbestosis.

[Modifications of the cytoimmunological pattern of bronchoalveolar lavage (BAL) material caused by cigarette smoking in selected lower air-way diseases]. / Modyfikacje obrazu cytoimmunologicznego materialu z plukania oskrzelowo- pecherzykowego (bronchoalveolar lavage, BAL) powodowane przez palenie papierosów w wybranych chorobach dolnych dróg oddechowych.

INTRODUCTION: In the last years we have used flow cytometry as an auxiliary diagnostic tool in alveolar lymphocyte (i.e. originating from BAL) phenotyping in more than 500 persons suspected for lower airways pathology. MATERIAL AND METHODS: In the study we compared the results of 1) BAL lymphocyte typing by flow cytometry, 2) cytological examination, respectively, in nonsmoking/smoking (NS/S) patients with lung sarcoidosis, n = 56/31, extrinsic allergic alveolitis (EAA), n = 9/5, silicosis, n = 15/18, idiopathic pulmonary fibrosis (IPF), n = 20/7, and pulmonary tuberculosis (TBC), n = 7/6. The results were related to the volume of BAL fluid recovery (higher value reflects the dominance of lower airways content versus bronchial content). RESULTS: In smoking patients, in comparison with respective NS, significantly higher total BAL cell numer (except TBC), higher macrophage percentage, lower lymphocyte percentage and lower CD4/CD8 ratio (except EAA) was found. CD4/CD8 results: 8.26 +/- 0.52 (NS) vs 4.29 +/- 0.65 (S) in sarcoidosis (p < 0.001), 1.18 +/- 0.44 (NS) vs 0.99 +/- 0.43 (S) in IPF (p < 0.05), 1.79 +/- 0.22 (NS) vs 0.73 +/- 0.11 (S) in silicosis (p < 0.001) and 1.64 +/- 0.57 vs 0.88 +/- 0.1 in TBC (p < 0.05). Additionally, cigarette smoking modified BAL pattern: 1. in sarcoidosis and silicosis lower CD4+ cell and higher CD8+ cell percentage; 2. in IPF increase in neutrophil percentage; 3. in TBC higher neutrophil and eosinophil percentage. Both in NS and S, BAL fluid recovery rate is significantly positively correlated with CD4/CD8 ratio and total BAL CD3+ cell number and negatively with BAL CD8+ cell percentage. CONCLUSIONS: Interpreting of BAL material cytoimmunology pattern should take into account data on cigarette smoking and BAL fluid recovery rate. The results obtained in the study may reflect more severe disease course in IPF and TBC.

[Interferon gamma (IFN-gamma) level in broncholaveolar lavage (BAL) fluid is positively correlated with CD4/CD8 ratio in selected interstitial lung diseases]. / Stezenie interferonu gamma (IFN-gamma) w plynie z plukania oskrzelowo-pqcherzykowego w wybranych chorobach sródmiaqszowych pluc--dodatnio skorelowane z wartoscia stosunku CD4/CD8.

UNLABELLED: IFN-gamma a potent antifibrotic activity in interstitial lung diseases (ILD). T cells, both Th1 and Tc1, are considered to be the main local source of IFN-gamma. MATERIAL AND METHODS: BAL fluids of 98 patients with ILD, incl. idiopathic pulmonary fibrosis (IPF/UIP), sarcoidosis, extrinsic allergic alveolitis (EAA), asbestosis and silicosis (n=16, 49, 7, 10, 16 resp.) were tested with ELISA for IFN-gamma levels. Results were compared with BAL cytoimmunology and patients' clinical data. RESULTS: Significantly increased IFN-gamma levels were found in non-treated patients with EAA (7.8 +/- 2.1), IPF (6.1 +/- 1.8), Loefgren's syndrome, LS (11.9 +/- 2.6) and progressive sarcoidosis, PS (6.4 +/- 1.2, p < 0.05 for all), whereas the results in pneumoconioses were comparable to those obtained in controls (2.0 +/- 1.1 pg/ml, median +/- SEM). IFN-gamma results were positively correlated with total number of CD4+ cells (r(s) = +0.38, p < 0.05), CD4+ cells percentage (r(s) = +0.32, p < 0.005) and CD4+/CD8+ ratio (r(s) = +0.38, p = 0.0007), but negatively correlated with CD8+ cell percentage (r(s) = -0.39, p < 0.0005). In IPF patients with CD4/CD8 < or =1 (n=9) IFN-gamma level was lower as compared with the group with CD4/CD8 >1 (n=7), 2.8 +/- 1.3 vs. 7.3 +/- 1.0 pg/ml. In sarcoidosis, IFN-gamma level did not seem to have a prognostic role, since values obtained in PS did not differ remarkably from those in stable sarcoidosis and LS. Moreover, subsequent steroid treatment in 7 patients with progressive sarcoidosis did not change significantly IFN-gamma levels in BAL fluid. CONCLUSIONS: Increased IFN-gamma level was found in non-treated patients with IPF, Loefgren's syndrome and progressive sarcoidosis. CD4+ (Th1), but neither CD8+ (Tc1) nor NK cells seem to be the main local source of IFN-gamma in ILD. Relatively low CD4/CD8 ratio in ILD may indicate the patients with increased risk of lung fibrosis.

[Cigarette smoking results in the number of CD8+Fas Ligand+ T cytotoxic lymphocytes in bronchoalveolar lavage (BAL) fluid of patients with idiopathic pulmonary fibrosis (IPF)]. / Palenie papierosów zwieksza liczbe limfocytów cytotoksycznych CD8+Fas Ligand+ w plynie z plukania oskrzelowo-pecherzykowego (BAL) chorych z samoistnym wlóknieniem pluc (IPF).

Idiopathic pulmonary fibrosis (IPF) is an interstitial lung disease with unfavourable outcome. Tobacco consumption in IPF exacerbates the clinical manifestations and limits the time of patient survival. The cyto-immunological alterations caused by smoking in IPF patients need particular explanation. BAL was carried out in 21 non-treated patients with IPF, subdivided according to the smoking status (n=7 for smokers). BAL routine cytology was completed by; immunotyping, including T cell major subsets (CD4 and CD8) stained for Fas, Fas ligand (FasL) and TNFR-1, late apoptosis/cell cycle analyses (BAL cells were permeabilized and stained with PI) and TUNEL assay. BAL cytology in IPF, as compared with control group, was characterized by significantly higher total cell and macrophage number, increased lymphocyte, neutrophile and eosinophile percentage and relatively low CD4/CD8 ratio. Cigarette smoking in IPF resulted in enhanced BAL lymphocyte CD8 cell percentage and number, as compared with nonsmoking subgroup and further decline in CD4/CD8 ratio (0.41+/-0.15 vs 1.23+/-0.29 in nonsmokers, median +/- SEM, p<0.05). The percentage of CD8, but not CD4 cells carrying Fas Ligand was significantly increased in IPF smokers (12.0+/-3.1% vs 3.7+/-0.9% in nonsmokers, median +/- SEM, p<0.05). Apoptosis rate of BAL macrophages and lymphocytes was enhanced in IPF, as compared with controls (confirmed by both techniques), but without remarkable changes, if compared one IPF subgroup to another. The number and percentage of CD8+FasL+ was negatively correlated with vital capacity (VC) values in IPF patients, but not with BAL inflammatory cell apoptosis rate. Cigarette smoking enhanced a percentage as well as a total number of both BAL CD8 and BAL CD8+FasL+ cells in IPF patients. BAL cytotoxic cells (CD8+FasL+ lymphocytes) seem to have impact on impaired lung function in smoking IPF patients.

Expression of insulin-like growth factor-I (IGF-I) in alveolar macrophages and lymphocytes obtained by bronchoalveolar lavage (BAL) in interstitial lung diseases (ILD). Assessment of IGF-I as a potential local mitogen and antiapoptotic cytokine.

Little is known about IGF-I expression in the alveolar lymphocytes (AL), and about local role of IGF-I in physiological conditions and in interstitial lung diseases. Bronchoalveolar lavage was carried out in patients with silicosis, asbestosis, idiopathic pulmonary fibrosis (IPF) and sarcoidosis, as well as in control subjects (n = 13, 9, 12, 56, 15, resp). Alveolar macrophages (AM) and lymphocytes (AL) were studied for (1) IGF-I, BCL-2, Fas and Fas Ligand expression and (2) cell cycle (incl. sub-G1 peak of late apoptosis) with propidium iodide (PI). Flow cytometry (FC) and immunocytochemistry were used. AL early apoptosis was detected by Annexin V FITC/PI staining. IGF-I was present in AL of all tested groups. The number of IGF-I positive AL was significantly higher in IPF (52 +/- 6.7%) and in later (II and III) stages of sarcoidosis (39 +/- 7.8 vs 16 +/- 4.0% in controls, p < 0.05). Increased BCL-2 expression in AL was detected in IPF and sarcoidosis. In all tested groups, AL were almost exclusively Fas+ T cells. Generally, a low number of AL entered apoptosis; no significant differences were found between patient groups, except decreased apoptosis rate in sarcoidosis (0.60 +/- 0.17 vs 1.15 +/- 0.33% in controls, p < 0.05). Proportion of AL positive for IGF-I was significantly correlated with parameters reflecting AL and AM cell proliferation and BCL-2 expression (e.g. AL IGF-I+ vs AM in S phase of cell cycle: r(S) = +0.50, p = 0.001), but not with apoptosis. The results show that human alveolar lymphocytes express IGF-I in normal conditions, as well as in ILD. The proportion of IGF-I+ lymphocytes was significantly increased in IPF and at later stages of sarcoidosis. In our material there was no evidence for profibrogenic or antiapoptotic activity of IGF-I. We suggest that IGF-I originating from AL may be locally active as a mitogen for alveolar macrophages and lymphocytes in ILD.

[Apoptosis of alveolar lymphocytes in sarcoidosis and in control group is more frequent in smokers than in nonsmoking persons]. / Apoptoza limfocytów pecherzykowych w sarkoidozie i w grupie kontrolnej wystepuje czesciej u palaczy papierosów niz u osób niepalacych.

Cigarette smoking enhances apoptosis rate of alveolar macrophages. However, little is known about the appearance and extension of apoptosis in bronchoalveolar lavage (BAL) lymphocytes originating from smoker individuals, both in pulmonary sarcoidosis (the disease characterized by lymphocytic alveolitis) and in controls. BAL was carried out in 60 nontreated patients with pulmonary sarcoidosis, subdivided acc. smoking status and in 22 control persons, free of any lung pathology. BAL (alveolar) lymphocytes were a) stained for TUNEL; b) permeabilized and stained with PI for late apoptosis/cell cycle analyses; c) immunophenotyped, including CD95, CD95 Ligand, Bcl-2, Bcl-XL, Bak and insulin-like growth factor-I (IGF-I) expression. BD FACSCalibure flow cytometer, PC Lysys and ModFit software were applied. The low number of AL entered apoptosis, which was confirmed by both techniques. Cigarette smokers were characterized by higher AL apoptosis percentage in respective subgroups (sarcoidosis: 0.6 +/- 0.13 in nonsmokers vs 0.9 +/- 0.23 in smokers; controls: 0.85 +/- 0.23 in nonsmokers vs 1.5 +/- 0.35 smokers, median +/- SEM, p < 0.05); the proliferation rate was lower. Decreased IGF-I expression in AL of sarcoidosis smokers was observed (13.5 +/- 9.2 vs 46.0 +/- 6.0 in nonsmokers, p < 0.05). No differences were found between studied groups in expression of Bcl-2, Fas and FasL molecules (except significantly declined ratio of CD8+FasL+ cells in sarcoidosis nonsmokers). AL apoptosis rate was positively correlated with respective alveolar macrophage results (Rs = +0.59, p < 0.00001) and negatively with CD4/CD8 ratio (Rs = -0.32, p < 0.001); no correlation was found with lung function test results and with Bcl-2, Fas and FasL expression in BAL cells. Apoptosis of alveolar lymphocytes was more frequent in nonsmokers both in pulmonary sarcoidosis and in controls; lower AL percentage proliferates. These phenomena seem to participate in lower AL percentage, observed in smoker subgroup of sarcoidosis. Some mechanisms of local apoptosis alterations in smokers may be common for alveolar lymphocytes and macrophages.

[Clinical course of acute poisoning with olanzapine]. / Kliniczny przebieg ostrego zatrucia olanzapina.

Olanzapine is a new atypical antipsychotic drug acting on different receptors. A variety of pharmacologic effects are responsible for toxicity and the variety of clinical symptoms seen in overdose: tachycardia, agitation or aggression, dysarthria, extrapyramidal dystonic effects, sedation or coma, small pupils, blurred vision, respiratory depression, hypotension. A retrospective analysis of clinical course of eight acute olanzapine intoxication treated at the Department of Clinical Toxicology Jagiellonian University Medical College is presented. CNS symptoms manifested in fluctuations between somnolence/coma and agitation/aggression and miosis were observed in most of the patients. Increased CPK activity was stated in the most of patients. All of the patients recovered, poisoning severity according PSS was moderate and severe.

Fly agaric (Amanita muscaria) poisoning, case report and review.

Gathering and eating mushrooms and other plants containing psychoactive substances has become increasingly popular among young people experimenting with drugs. Dried fly agaric Amanita muscaria fruiting bodies were eaten by five young persons (18-21 years of age) at a party in order to evoke hallucinations. Visual and auditory hallucinations occurred in four of them, whereas a 18-year-old girl lost consciousness. The following morning, she went to the Clinic of Toxicology. Due to the fact that not all the active substances present in the fly agaric have been identified, and some of them have an effect after a period of latency, the patient was admitted for several days of observation during which check-up examinations were performed. After four days without any problems, she was discharged. The poisoning regressed with no organ complications. The remaining persons who had eaten the fly agaric were free from any complaints.