DIAPH2 alterations increase cellular motility and may contribute to the metastatic potential of laryngeal squamous cell carcinoma.

Low 5-year survival rate in laryngeal squamous cell carcinoma (LSCC) is to large extent attributable to high rate of recurrences and metastases. Despite the importance of the latter process, its complex genetic background remains not fully understood. Recently, we identified two metastasis-related candidate genes, DIAPH2 and DIAPH3 to be frequently targeted by hemizygous/homozygous deletions, respectively, in LSCC cell lines. They physiologically regulate such processes as cell movement and adhesion, hence we found it as a rationale, to study if tumor LSCC specimens harbor mutations of these genes and whether the mutations are associated with metastasizing tumors. As a proof of concept, we sequenced both genes in five LSCC cell lines derived from lymph node metastases assuming there the highest probability of finding alterations. Indeed, we identified one hemizygous deletion (c.3116_3240del125) in DIAPH2 targeting the FH2 domain. Moreover, we analyzed 95 LSCC tumors (53 N0 and 42 N+) using the Illumina platform and identified three heterozygous single nucleotide variants in DIAPH2 targeting conserved domains exclusively in N+ tumors. By combining these results with cBioPortal data we showed significant enrichment of DIAPH2 mutations (P = 0.036) in N+ tumors. To demonstrate the consequences of DIAPH2 inactivation, CRISPR/Cas9 editing was used to obtain a heterozygous DIAPH2+/- mutant HEK-293T cell line. Importantly, the edited line shows a shift from 'proliferation' to 'migration' phenotype typically observed in metastasizing cells. In conclusion, we report that DIAPH2 alterations are present primarily in metastasizing specimens of LSCC and suggest that they may contribute to the metastatic potential of the tumor.

gamma delta-Lymphocytes in the peripheral blood of patients with rheumatoid arthritis--correlation with clinical and laboratory parameters of the disease and with the treatment used.

Rheumatoid arthritis (RA) is a chronic, autoimmune disease in the pathogenesis of which T-lymphocytes are believed to play a crucial role. The aim of this work was to investigate a subpopulation of gamma delta-lymphocytes in the peripheral blood of patients with RA in the context of chosen parameters of the disease. Moreover, we studied the influence of disease modifying antirheumatic drugs (DMARDs) on the number of gamma delta-lymphocytes in the peripheral blood of patients with RA. The gamma delta-lymphocytes were studied using monoclonal anti-delta 1 antibodies labelled with fluorescein (Becton Dickinson) then counted under a fluorescence microscope (Olympus). We found a significant increase in the number of gamma delta-lymphocytes in the peripheral blood of patients with RA as compared with normal controls. The highest numbers were observed in patients with RA and extraarticular manifestation of the disease. Treatment with DMARDs led to a gradual normalisation of the elevated values of gamma delta-lymphocytes.

[The fiftieth anniversary of Miodonski's electro-rhino-spirometry]. / Piecdziesieciolecie elektrorynospirometrii Miodonskiego.

On the 50th anniversary of the introduction of the electro-rhino-spirometry by Miodonski the author recalls its theoretical foundations and diagnostic possibilities. Moreover, it has been stressed that Miodonski's method was the first one which does not disturb the nasal function, and the development of electronics supported and widened its diagnostic possibilities leaving the theoretical basis for examination unchanged.

Does chemotherapy-induced leukopenia predict a response in small-cell lung cancer?

The correlation between chemotherapy-induced toxicity and treatment outcome in cancer patients has not been studied thoroughly. Our aim was to evaluate whether there is any relationship between chemotherapy-induced leukopenia and response to treatment in small-cell lung cancer (SCLC). Data derived from records of 228 patients treated within two prospective multicentre phase II studies were analysed. In the first study (101 patients) chemotherapy included vincristine, epirubicin and cyclophosphamide and, in the second (127 patients), cyclophosphamide, etoposide and epirubicin; both regimens were given every 3 weeks. In the present analysis, the correlation between treatment outcome (response rate and survival) and highest scores of leukopenia within the first two and up to the fourth chemotherapy cycle, respectively, was evaluated. The objective response rate for the entire group was 66%; 53% in patients whose white blood cells remained normal and 85% in those who developed leukopenia within the first two cycles (P = 0.000). In multifactorial analysis, also including other treatment- and patient-related factors, independent correlation with response to chemotherapy was found for leukopenia (P = 0.001), chemotherapy regimen (P = 0.002) and the combined relative dose intensity (P = 0.018), but not for patient sex, age, performance status, pre-study weight loss, extent of disease and initial white blood cell count. Leukopenia within the first two cycles of chemotherapy was not correlated with survival, whereas such correlation for leukopenia occurring up to the fourth cycle was at the borderline level (P = 0.06). These findings suggest a relationship between chemotherapy-induced leukopenia and tumour response in SCLC.

TNF-alpha, IL-1 and IL-6 concentration in bronchoalveolar lavage fluid (BALF) of non-small cell lung cancer (NSCLC).

Fiberoptic bronchoscopy and bronchoalveolar lavage (BAL) provide facilities for biologically active substances directly produced by the tumor. In the present study we have investigated the concentration of the following cytokines: TNF-alpha, IL-2 and IL-6 in bronchoalveolar lavage fluid (BALF) of non-small cell lung cancer (NSCLC) before treatment. The study group consisted of 20 patients with squamous cell carcinoma (Gr. I). The control group consisted of 18 patients with non-malignant lung disease (6 patients with sarcoidosis and 12 with COPD). All patients underwent bronchoscopy followed by bronchoalveolar lavage (BAL). The concentrations of the above mentioned cytokines were measured using Sorin's ETI system for EIA analysis. Statistical analysis was performed within the groups, between the groups, and in different stages of malignant disease. The mean TNF-alpha concentration in Gr. I was 1192 pg/ml/mg p. and was significantly higher than in sarcoidosis (5.3 pg/ml/ mg p.) and COPD (0.5 pg/ml/mg p.). We observed a correlation between TNF-alpha concentration and the stage of malignant disease. The highest concentration was in IIIb stage (2150 pg/ml/mg p.). IL-6 concentration in malignant patients was strongly correlated with TNF-alpha concentration and was significantly higher than in control (265.868 pg/ml/mg p. in cancer patients, in sarcoidosis: 21.694 pg/ml/mg p. and in COPD: 40.708 pg/ml/mg p.). It was the highest in stage II (334 pg/ml/mg p.). IL-1 concentrations were not significantly higher in malignant patients (50.173 pg/ml/mg p., nor in IIIa stage (70.136) pg/ml/mg p. as compared with controls (18.648 pg/ml/mg p. in sarcoidosis and 28.395 pg/ml/mg p. in COPD).

IL-2 concentration in bronchoalveolar lavage fluid (BALF) of non-small cell lung cancer (NSCLC) patients.

The results of treatment and the survival time of lung cancer patients are strictly dependent on early diagnosis. Fiberoptic bronchoscopy and bronchoalveolar lavage (BAL) are the most effective diagnostic methods in cancer diagnosis. These methods allowed us to evaluate biological neoplastic markers at the site of the tumor. Using commercially available ELISA kits (Endogen) followed by ETI-system (SORIN) analysis we measured the IL-2 concentration in BALF of 36 non-small cell lung cancer (NSCLC) patients before (Gr. I) and after (Gr. II) surgery and 18 non-neoplastic lung disorder patients as a control group (6 cases of sarcoidosis and 12 cases of COPD). In BALF of Gr. I Macrophage percentage was higher (x = 74.125%), and lymphocyte percentage was lower (x = 15.875%) than in sarcoidosis x = 38.33% and x = 64.3% respectively. IL-2 of BALF was not detected in 83.4% of squamous cell lung cancer cases (Gr. I) before treatment. The average IL-2 BALF concentration of the remaining portion of this group was 80.49pg/ml/mg p. IL-2 was detected in Gr. II (x = 151.003 pg/ml/mg p.) after combined cancer resection. An inverse correlation was found between IL-2 BALF concentration and disease stage.

[The sixtieth anniversary of Miodonski's paraboloid head lamp reflector]. / Szescdziesieciolecie czolowego reflektora parabolicznego Miodonskiego.

On the 60th anniversary of the elaboration by Miodonski of the optical solutions and his construction of the Paraboloid Headlamp and its application to clinical practice, the author compares its characteristics with other types of lamp used in otolaryngology on the example of Clar's Headlamp and Headmirror. This comparison shows that even now Miodonski's Paraboloid Headlamp due to its qualities, has no equal. The merits of Miodonski's Headlamp are as follows: 1. Permits a change of the axis of illumination under sterile condition; 2. Gives the most powerful light; 3. Permits binocular inspection of very narrow and long canals and ducts allowing for enlargement from the time that Miodonski added a magnifying and mobilizing lens to his headlamp in 1961.

[Level of D-dimer and comparison with cell count in pleural effusion from tuberculosis and neoplasms]. / Stezenie D-dimeru w porównaniu z liczba komórek w plynach oplucnowych pochodzenia gruzliczego i nowotworowego.

The examinations were carried out in 33 patients (aged 21-69 years) with tuberculous and neoplastic effusions, before and during the treatment. 10 subjects with circulatory failure and pleural effusion a consisted control group. D-dimer level was detected using semiquantitative method (D-dimer Latex Test, Diagnostica Stago-Boehringer Mannheim), cells number and composition-using light microscope. All the groups demonstrated significant differences in the effusion D-dimer level before and during the treatment. Cells number was significantly higher in tuberculous and carcinomatous pleural effusion than in the control group. D-dimer level correlated with neutrophils percentage only in tuberculous effusions before the treatment. We found no correlation in D-dimer level and cells number and composition in neoplastic pleural effusion. We did not find any correlation in D-dimer level and cells number in blood.

[Selective increase of gamma delta T lymphocytes in peripheral blood of patients with pulmonary tuberculosis according to the kind and stage of disease]. / Wybiórczy wzrost liczby limfocytów gamma delta T we krwi obwodowej chorych na gruzlice pluc w róznej postaci i fazie choroby.

The aim of this study was to evaluate the distribution of gamma tau T cells expressing gd T cell receptor (TCR) in the peripheral blood of 42 patients with pulmonary tuberculosis before, and during the treatment. Control groups were 15 healthy individuals and 17 patients with non granulomatous diseases. In these groups number of g/d T cells were 240 and 239 cells/ml and percentage 9.8 and 12.3, respectively. Using direct immunofluorescence with the anti-gamma delta TCR monoclonal antibodies followed by microscopy analysis, the total number and the percentage of gamma delta T cells in purified peripheral blood mononuclear cells (PBMC) was analyzed. Expression of CD4, was determined. The observation period amounted to 12 weeks of the initial treatment. The total number of gamma delta TCR in blood of tuberculous patients was 704 cells/ml and 40% respectively, and normalized during the treatment. An inverse correlation was found with the proportion of CD4+ cells in blood.

[Morphology of mast cells in experimental pulmonary fibrosis induced with bleomysin]. / Morfologia komórek tucznych w doswiadczalnym wlóknieniu pluc wywolanym przez bleomycyne.

Bleomycin induces local inflammatory process with subsequent pulmonary fibrosis. An unknown chemoattractant induces the mast cell (MC) migration to the injured lung tissue. Aim of this study was evaluation of the number, topography and ultrastructure (TEM) of the MC in rat lungs with bleomycin-induced fibrosis. The bleomycin was administered once intratracheally (3.6 mg/kg). The animals were sacrificed on 7th. 14th and 21st day of experiment. MC were stained with Csaba's method. An evident increase in MC number related to the phase of experiment and stage of lung fibrosis was observed: 520, 1200, 4745 per cm2 section on the 7th, 14th and 21st day respectively. In control the MC number was 163 per cm2 section (p < 0.001). On the 7th day the MC were rich in red granules (mature granules with preponderance of heparin). They were located mainly in pleura and around the blood vessels, as in control. On the 14th and 21st day the majority of MC was situated in places of active fibrosing. They contained exclusively blue granules (the young granules with preponderance of biogenic amines), mixture of increased secretory function of the MC in the fibrotic lungs. Some of the MC granules showed fusion and altered matrix contents, other were emptied in piecemeal manner. A net of microtubules connected the granules was observed. Degranulation of MC may release heparin and cytokines able to stimulate synthesis of extracellular matrix. It has been suggested that heparin contributes to the fibrotic process and angiogenesis, stimulating directly or indirectly collagen synthesis by binding, stabilization and activation of fibroblast growth factor (FGF) and cell adhesion glycoproteins.

[Analysis of prognostic factors for long term survival in patients with small cell lung cancer]. / Analiza czynników rokowniczych warunkujacych dlugotrwale przezycia chorych na drobnokomórkowego raka pluca.

Prognostic factors for long term survival were analyzed in a group of 719 patients with small cell lung carcinoma treated within 4 consecutive prospective multicenter trials between 1981 and 1990. 74 patients (10.3%) survived more than 2 years; 30 of them (4.2%) with no evidence of disease. The most significant determinator of prolonged survival was extent of disease: 13.9% (59/424) of patients with limited disease vs. 5.1% (15/295) with extensive disease survived more than 2 years (p < 0.001). Of 138 female patients 24 (17.4%) were long term survivors, compared to 8.6% (50/581) of males (p < 0.01). Initial good performance status and no weight loss were also found to be correlated with long term survival. Of the group of 2-year survivors 51 patients subsequently died (median survival duration 31 months), 10 are alive with cancer or are lost to follow up and 13 are in complete remission with median follow up of 64 months. 20 patients (3.8%) survived more than 5 years. This study confirms the possibility of cure in SCLC, especially in patients with favorable prognostic factors.

Usefulness of a multiple biomarker assay in bronchoalveolar lavage (BAL) and serum for the diagnosis of small cell lung cancer.

To evaluate the diagnostic usefulness of simultaneous determinations of carcinoembryonic antigen (CEA), neuron specific enolase (NSE), chorionic gonadotrophin (HCG) and carbohydrate antigenic determinant 19-9 (CA 19-9), we studied 48 patients with small cell lung carcinoma (SCLC) and 15 with nonmalignant lung disease. The combination of CEA-BAL, NSE-BAL, and NSE-serum taken together with results of bronchoscopy (histologic and cytologic) showed the highest discriminating power between malignant (SCLC) and nonmalignant lung disease. The sensitivity of bronchoscopy alone was 35%. However, when bronchoscopy results were combined with 3 positive markers, the sensitivity increased to 71%, with at least 2 positive markers to 94%, and with at least 1 positive marker to 100%. When both bronchoscopy and all 3 markers were negative, the results showed a negative predictive value of 100%.

Value of CEA and SCC-Ag in bronchoalveolar lavage (BAL) and serum of patients with lung carcinoma.

To evaluate the diagnostic usefulness of simultaneous determinations of carcinoembryonic antigen (CEA) and squamous cell carcinoma antigen (SCC-Ag), we studied 25 patients with lung carcinoma and 12 with nonmalignant lung diseases. The measurements were made in serum and bronchoalveolar lavage (BAL). The results showed that positive rates with lavage CEA and SCC-Ag in lung carcinoma patients were higher in comparison with those markers in serum. The combination of lavage CEA and SCC-Ag taken together with the results of bronchoscopy (histology and cytology) showed the highest discriminating power between malignant and nonmalignant lung diseases. The sensitivity of bronchoscopy increased from 48 to 84% with at least 1 positive marker. It appears that the simultaneous determination of CEA and SCC-Ag levels in serum and BAL in lung carcinoma patients may be of considerable importance in diagnosis.

Combination chemotherapy with vincristine, epirubicin and cyclophosphamide in small cell lung carcinoma. Polish Lung Cancer Cooperative Group.

The aim of this prospective study was to assess the activity of a combination of vincristine, epirubicin and cyclosphosphamide (VEC) in previously untreated patients with limited small cell lung carcinoma (SCLC) and to delineate the feasibility of dose escalation for epirubicin in this regimen. The chemotherapy schedule included cyclophosphamide, 1000 mg/m2, vincristine, 1 mg/m2 and escalating doses of epirubicin: 50 mg/m2, 70 mg/m2 and 90 mg/m2; respectively in three consecutive groups of patients. Drug cycles were repeated every 3 weeks. 118 patients from eight institutions were enrolled in this study between February 1986 and March 1989. Objective tumour response was observed in 81 of 116 evaluable patients (70%) including 25 patients (22%) who achieved a complete remission. Responding patients received thoracic radiation after the fourth cycle of chemotherapy. The median duration of response was 30 weeks and the median duration of survival was 52 weeks. There were no significant differences in treatment results between the consecutive groups of patients. The regimen was well tolerated for all doses of epirubicin. The main toxicities included alopecia (96%), nausea and vomiting (81%) and leukopenia (44%). Grade 4 haematological toxicity was observed in 3 patients (2.6%). No significant epirubicin dose-dependent side effects, except for mucositis were observed.