Clinical validation of grass pollen exposure chamber in patients with allergic rhinitis triggered by timothy grass.

BACKGROUND: The fluctuation in concentrations of airborne allergens frequently presents a challenge to assessing the efficacy of allergen immunotherapy (AIT) in 'field' studies. Allergen exposure chambers (AECs) are specialized medical installations developed to expose individuals to allergens at defined and consistent concentrations under a controlled environment. The aim of the study was to validate the provocation test with timothy grass pollen as well as to assess its safety in the AEC in patients with allergic rhinitis. METHODS: In the ALLEC® AEC, varying concentrations of timothy grass pollen were dispersed. Allergic symptoms were measured by total nasal symptom score (TNSS), acoustic rhinometry, peak nasal inspiratory flow (PNIF) and nasal discharge volume. Lung function, assessed through peak expiratory flow rate (PEFR) and forced expiratory volume in the first second (FEV1), was used to evaluate safety. RESULTS: The consistency of the test was proved by the stability of environmental conditions, including temperature, humidity and CO2 levels, as well as constant concentrations of grass pollen at predetermined levels ranging from 1000 to 10,000 particles per cubic meter (p/m3). Allergic individuals developed symptoms at concentrations of 3000 p/m3 and above, across all measured endpoints. Lung function was not affected throughout all the challenges. The reproducibility of symptoms was confirmed throughout the tests. The concentration of 8000 p/m3 together with a challenge duration of 120 min was found to be optimal. CONCLUSION: The study demonstrates that the ALLEC® grass pollen exposure chamber provides a reliable and safe method for inducing repeatable symptoms in patients with allergic rhinitis. This approach can be effectively applied for allergy diagnostics and clinical endpoint determination during AIT.

[Expression of p53, bcl-2 and nm23 proteins in squamous cell lung cancer]. / Ekspresja bialek p53, bcl-2 i nm23 w plaskonablonkowym raku pluca.

INTRODUCTION: The evaluation of lung cancer molecular profiles is an essential element of the therapeutic process in that type of neoplasm. The analysis of apoptotic and metastasis-linked proteins is an important goal because of the key role of those processes in carcinogenesis. The aim of this study was to evaluate the expression of apoptosis regulatory proteins p53 and bcl-2 as well as antimetastatic marker nm23 in squamous cell lung cancer, taking into account the clinical and pathological data. MATERIAL AND METHODS: Thirty tissue specimens from patients undergoing therapeutic or diagnostic thoracic surgery were included in the study. All markers were assessed with immunohistochemistry method on paraffin-embedded tissue. RESULTS: Nm23 expression was observed less frequently in specimens with cancer cell emboli in blood vessels or lymph node metastasis. In cancers with lymph node metastasis, the coexpression of p53 and bcl-2 was found statistically more often than in lymph node negative cases. There was no correlation between p53, bcl-2 and nm23 expression and 2-years survival time. CONCLUSIONS: Our study indicates a marked heterogeneity of p53, bcl-2 and nm23 expression in squamous cell lung cancer and the potentially unfavorable influence of p53 and bcl-2 coexpression. Less frequent nm23 expression seems to be connected with morphological signs of metastatic process.

Apoptotic markers p53, Bcl-2 and Bax in primary lung cancer.

BACKGROUND: Apoptosis is the fundamental process necessary for eliminating damaged or unwanted cells. Alterations in the apoptotic pathway appear to be key events in cancer development and progression. The aim of the study was to determine the p53, Bcl-2 and Bax expressions in lung cancer, taking into account histological heterogeneity and the adjacent bronchial resection margin. MATERIALS AND METHODS: Tissue specimens from 60 histopathologically verified lung cancer specimens and 12 bronchial stumps were evaluated. The presence of the studied markers was revealed by immunocytochemistry on paraffin-embedded tissue. RESULTS: The percentage of p53- and Bax-positive lung cancers was comparable (51.6% for both proteins), while Bcl-2 immunoreactivity was observed in fewer (31.6%) cases. There was no p53 accumulation in bronchial stumps, while Bcl-2 and Bax staining formed a repeatable specific pattern in bronchial epithelium. The differences in apoptotic marker expression between non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) were revealed, especially regarding p53 and Bax expression (60% vs. 10%, p = 0.005 and 58% vs. 20%, p = 0.04, respectively). Taking into account the histological structure of NSCLC, Bax expression appeared to be more frequent in adenocarcinoma than in squamous cell lung cancers (88% vs. 42%, p = 0.004). No interrelationship between the studied proteins in lung cancer tissue was revealed. CONCLUSION: The expression of p53, Bcl-2 and Bax was altered in lung cancer tissue compared to histologically normal bronchial epithelium. The difference between apoptotic marker expression in NSCLC and SCLC could reflect the different pathogenesis of these two lung pathologies.

Temperature-sensitive ovarian carcinoma cell line (OvBH-1).

OvBH-1 cells from a patient with ovarian clear cell carcinoma were established and their biochemical status was analyzed. Cells grown at 37 degrees C exhibited normal cell cycle distribution, whereas the cells shifted to 31 degrees C were arrested in the G(2) / M phase of the cell cycle. Immunochemical analysis using anti-p53 antibodies (DO-1, PAb240, PAb421, and PAb1620) revealed that only the DO-1 antibody reacted with p53 with a high and similar percentage at both temperatures. PAb240 reacted with a low percentage of cells at 37 degrees C and no reaction was observed at 31 degrees C. PAb421 antibody stained a significantly lower percentage of cells at 37 degrees C than at 31 degrees C. Cells were not stained with PAb1620 antibody and were negative for antibodies against p21(WAF1) and MDM2 proteins independently of the temperature. Sequencing of all coding exons of the p53 gene demonstrated only a neutral genetic polymorphism, i.e. a G-to-A substitution (GAG to GAA) at nucleotide position 13 432. Thus, the observed temperature sensitivity of OvBH-1 cells cannot be ascribed to a p53 primary structure mutation. Based upon immunochemical analyses, we consider, however, that p53 in nuclei of OvBH-1 cells is in a highly unstable conformation. Furthermore, the N-terminal portion of the p53 protein at Ser20 has not been modified, and Lys373 and / or Ser378 of the C-terminus is acetylated and / or phosphorylated. The nuclear location signal of p53 is preserved. Induction of MDM2 protein is uncoupled from the cell regulatory machinery and the induction of p21(WAF1) by p53 is impaired in OvBH-1 cells.