Evaluation of fluorescence-based methods for total vs. amplifiable DNA quantification in plasma of lung cancer patients.

In the last decade numerous reports demonstrated that free-circulating DNA in plasma/serum samples might be a promising biomarker in a number of pathologies, including cancer. Thus, choosing the reliable and efficient method of plasma DNA quantification would be an essential step prior to any clinical evaluation of cell-free DNA measurement in cancer patients. The aim of present study was to compare two highly-sensitive DNA quantification methods in regard to their applicability and effectiveness in monitoring the cell-free DNA level in the blood of patients with resectable non-small cell lung cancer. Plasma samples collected from 10 patients before any treatment, after neoadjuvant therapy and subsequent surgery, were used for DNA quantification by direct fluorescent PicoGreen staining and by real-time qPCR in SYBR Green and TaqMan probe approach using beta-actin gene as the amplifying target. The PicoGreen method demonstrated a high level of correlation with both the SYBR Green (r=0.87, P<0.0001) and TaqMan probe approach (r=0.94, P<0.0001). The total DNA content, determined by PicoGreen, proved to be several-fold higher than the amplifiable DNA amount measured by real-time qPCR. Consequently, intercalating fluorochromes, like PicoGreen, might serve as a rapid, accurate, and inexpensive alternative to real-time qPCR for routine dsDNA quantification and multicenter standardization.

p53 and HER2/neu expression in relation to chemotherapy response in patients with non-small cell lung cancer.

The aim of the study was to investigate a relation between p53 and HER2/neu expression in resected lung tumors and the response of those tumors to neoadjuvant chemotherapy. The study population included 67 consecutive patients with non-small cell lung cancer (NSCLC) in stage II or III who were operated on at the Institute of Tuberculosis, Warsaw, Poland, between 20 April 2001 and 10 March 2003. All patients received two cycles of chemotherapy consisting of cisplatin and vinorelbine prior to the operation. The response to therapy was assessed as complete response (CR), partial response (PR), stable disease (SD) or progressive disease (PD), on the basis of CT scans performed before and after neoadjuvant chemotherapy. p53 and HER2/neu protein expression were evaluated by immunohistochemistry (IHC) using antibodies against p53 (clone PAb 1801, Novocastra) and against HER2/neu (Dako) in paraffin-embedded specimens of tumors. A response to therapy (CR+PR) was observed in 27 patients, while 40 patients (SD+PD) were regarded as resistant to therapy. Resistance was observed significantly more often in tumors above 3 cm in diameter. p53 expression was found in 16 tumors (23.9%) and HER2/neu in 26 tumors (38.8%). We observed a nonsignificant tendency to chemoresistance in tumors with HER-2/neu overexpression and also in tumors with p53 overexpression. If we consider HER-2/neu and p53 together, chemoresistance was observed statistically significantly more often when one or both markers were positive (p<0.05). This significance was independent of tumor size.

Evaluation of treatment results and toxicity in cases of repeated radiation therapy of spinal metastasis.

The study evaluates retrospective results and toxicity in repeated radiation therapy in patients with recurrent pain caused by backbone metastasis, having undergone previous radiotherapy in the same body region. Fifty-seven patients were analyzed: 24 women and 33 men, aged 45-74 years (median = 59 years). They underwent a second radiation therapy treatment of the spinal column, between March 2002 and May 2004, performed due to recurrent pain in the previously irradiated region. The radiation used cobalt isotope 60 ((60)Co), to include the metastatically changed vertebrae and the margin of the adjusting healthy upper and lower vertebra. The radiated skin area measured 84-104 cm(2). Patients were divided into 3 groups depending on their treatment schemas: 12 patients -- first course of radiotherapy 4 Gy x 5, second 4 Gy x 5; 16 patients -- first course of radiotherapy 4 Gy x 5, second 8 Gy x 1; 29 patients -- first course of radiotherapy 8 Gy x 1, second 8 Gy x 1. The time delay between the first and the second radiation therapies was between 11 and 766 days (median = 135 days). An analgesic effect was achieved with most treated patients -- 41/57 (71.9%) with the use of second radiotherapy and with an insignificant percentage of complications, unimportant from the clinical point of view. No serious complications such as paralysis, paresis, spinal cord necrosis, neurological dysfunction of urethral or sigmoidorectal sphincters were noted in any of the treated patients. Based on our experience, this retrospective analysis shows usefulness of the second radiotherapy treatment as a safe method of palliative treatment in cases of painful bone metastasis appearing after a previous radiation therapy.

Lung cancer in women: age, smoking, histology, performance status, stage, initial treatment and survival. Population-based study of 20 561 cases.

BACKGROUND: The community-based cancer registry was set up and results were analysed to assess differences in clinicopathological parameters between women and men. PATIENTS AND METHODS: The Pulmonary Outpatient Departments supplied data on 20 561 lung cancer patients diagnosed in Poland from 1995 to 1998. Data regarding demographics, smoking, histology, treatment and survival were obtained. RESULTS: There were 2875 women and 17 686 men with lung cancer. Women were younger than men (60.02 versus 62.18 years; P <0.001). Age <50 years was more frequent in women than in men (23.3% versus 12%; P <0.001). Women with small-cell lung cancer (SCLC) and adenocarcinoma were significantly younger than women with squamous cancer (58.2 and 58.2 versus 61 years; P = 0.05). Also, men with adenocarcinoma and SCLC were younger than men with squamous cancer (60.6 and 60.2 versus 62.3 years; P = 0.05). Squamous cancer was the predominant type of lung cancer both in women (32.5%) and men (55.2%). However, SCLC (26.6% versus 19.9%: P <0.001) and adenocarcinoma (21.6% versus 9.6%; P <0.001) were more frequent in women than in men. Women were more frequently non-smokers than men (18.8% versus 2.4%; P <0.001). Adenocarcinoma patients smoked less intensively than patients with squamous and SCLC both in women (31.4 versus 35.8 and 33.7 packs/year; P <0.02) and in men (38.2 versus 42 and 41.9 packs/year; P <0.002). In multivariate analysis, bad performance status, advanced stage, non-surgical treatment, age >50 years at diagnosis and male gender were significant independent negative prognostic factors. CONCLUSIONS: Lung cancer was six times more frequent in men than in women. Women with lung cancer were younger than men and smoked less intensively. Over-representation of adenocarcinoma and SCLC was observed in the women. Women with lung cancer had a better prognosis than men.

Lung cancer in patients under 50 years old.

PURPOSE: The community based lung cancer registry was set up and the results were analysed to assess the differences in clinicopathological parameters and survival between patients under and over 50 years of age. PATIENTS AND METHODS: The Pulmonary Outpatient Clinics supplied the data on 5404 lung cancer patients diagnosed in Poland in 1995. Data regarding demographic, smoking, histology, clinical stage, performance status, family history of cancer, therapy and survival were obtained. RESULTS: At time of diagnosis 757 (14%) patients were under 50 years of age. In this group the frequency of females was higher as compared to this in the group of older patients (24.2% vs. 12.1%; P<0.001). Also the incidence of adenocarcinoma (12.6% vs. 7.6%; P<0.001) and small cell lung cancer (22.9% vs. 14.8%; P<0.001) were significantly higher in younger patients. Young patients had better performance status (55.4% vs. 46.6%; P<0.001) than old. The incidence of cancer in families of younger patients was higher both among the mothers (4.7% vs. 3.0%; P<0.001) and among the fathers (7.6% vs. 4.1%, P<0.001). Surgery or chemotherapy were more often applied to patients under 50 years in comparison to older ones (P<0.001). Young patients had better prognosis. Higher percentage of them survived one year (32.6% vs. 28.9%; P<0.049). In multivariate analysis, age over 50 at diagnosis, male gender, diagnosis of small cell lung cancer, advanced stage of the disease, bad performance status, and non-surgical therapy were independent negative prognostic factors. CONCLUSION: Among young patients, overrepresentation of women, subjects with positive family history of cancer, with better performance status, with adenocarcinoma and small cell lung cancer were noticed. Young patients were treated more aggressively and had better prognosis than patients over 50 years of age.

8-Oxo-7,8-dihydroguanine and 8-oxo-7,8-dihydro-2'-deoxyguanosine levels in human urine do not depend on diet.

In the present study, we used the method involving HPLC pre-purification followed by gas chromatography with isotope dilution mass spectrometric detection for the determination of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) and 8-oxo-7,8-dihydroguanine (8-oxoGua) in human urine. The mean levels of 8-oxoGua and 8-oxodGuo in the urine samples of the subjects on unrestricted diet were respectively 1.87 nmol/kg 24 h (+/-0.90) and 0.83 nmol/kg 24 h (+/-0.49), and in the case of the groups studied, they did not depend on the applied diet. The sum of the amounts of both compounds in urine can give information about the formation rate of 8-oxoGua in cellular DNA. It is also likely that the levels of modified nucleo-base/side in urine sample are reflective of the involvement of different repair pathways responsible for the removal of 8-oxodGuo from DNA, namely base excision repair (BER) and nucleotide excision repair (NER).

[Lung cancer--diagnosis and therapy delay]. / Rak pluca. Opóznienia w procesie diagnostyczno--leczniczym.

UNLABELLED: It is commonly known that in the course of neoplastic disease a diagnosis and therapy should be perform as fast as possible. It is particularly important for lung cancer patients. The goal of this study was to assess the diagnosis and therapy delay in unselected group of lung cancer patients, registered in Pulmonary Outpatients Clinics in all parts of Poland. MATERIAL: 20,561 lung cancer patients were registered in Pulmonary Outpatients Clinics in all parts of Poland from 1995 to 1998. RESULTS: The median delay caused by patients was about 46 days. In 33 provinces symptoms of the disease preceded diagnosis 28 to 50 days and in other 26 provinces--50 to 75 days. The median delay caused by doctors (time between first visit to the doctor and the date of diagnosis) was 65 days. In 35 provinces it was 30 to 70 days and in other 14 provinces this delay was between 70-111 days. The median time between first visit to the doctor and the beginning of therapy was 84 days. The median time between diagnosis and therapy was 30 days. Because chest physicians were also involved in the diagnosis and treatment of lung cancer patients, so for patients registered in years 1996-1998 the causes of delay connected with the function of this medical speciality were assessed. Median time between first visit to the doctor and first visit to the chest specialist was 38 days. Median delay to bronchoscopy was 26 days and to the diagnosis 46 days. CONCLUSION: Delay of diagnosis and therapy vary widely among different provinces of Poland. The delay generated by family doctors and chest physicians are very important and require a deeper evaluation on the province level in the future.

A multicenter, randomized, phase III study of docetaxel plus best supportive care versus best supportive care in chemotherapy-naive patients with metastatic or non-resectable localized non-small cell lung cancer (NSCLC).

This was an open-label randomized Phase III study of 207 patients with either unresectable or metastatic non-small cell lung cancer (NSCLC) who were treated with docetaxel plus best supportive care (BSC) or best supportive care alone. Patients in the chemotherapy arm of the study received docetaxel 100 mg/m(2) as a 1 h intravenous infusion every 21 days until they showed evidence of progressive disease, or estimated maximum benefit obtained or unacceptable side effects. Patients who received docetaxel were pretreated with oral dexamethasone. Patients in the BSC arm should not receive chemotherapy or anticancer therapy except for palliative radiotherapy. Overall survival obtained in the docetaxel arm was significantly longer than in the BSC arm (P=0.026). Two-year survival in the docetaxel arm was 12%, whereas none of the BSC patients survived after 20 months. The response rate was 13.1% (95% CI, 7.5-18.8%). There was a significantly longer time to progression in the docetaxel versus the BSC arm (P<0.001), and statistically significant improvement of clinical symptoms with docetaxel compared to BSC. The quality-of-life descriptors were in favor of docetaxel, and the difference was significant for pain, dyspnea and emotional functioning. The safety profile of docetaxel for this study was similar to that already reported in this patient population.

[The use of monoclonal antibodies in the detection of small cell lung cancer metastases in bone marrow]. / Próba zastosowania przeciwcial monoklonalnych do wykrywania przerzutów drobnokomórkowego raka pluca w szpiku.

Expression of a number of antigens associated with small cell lung cancer (SCLC) have been proposed as a marker of malignancy and the diagnostic tool for the staging procedures and important prognostic factor. Since the bone marrow (BM) was described as a frequent site for SCLC metastases, we have decided to assess clinical importance of cancer cells detection in BM, using immunofluorescence with MAC-1, MAC-31, NSE and anti-Fucosyl-GM1 (PF3) antibodies. The group of 32 patients with SCLC was assessed using our panel of antibodies. Control group consisted of 5 patients with other malignancies (3 patients with malignant lymphoma, 1 with chronic lymphocytic leukaemia and 1 with non-SCLC). The study revealed no correlation between the expression of SCLC markers in patients BM and the cancer treatment outcome measured as a response for treatment, time to progression, and survival time, and no significant difference was found between the patients and control group.

[Analysis of diagnostic errors and recommendations of diagnostic procedures in bacteriologically negative pulmonary tuberculosis]. / Analiza blednych rozpoznan u chorych na gruzlice czynna niepotwierdzona bakteriologicznie. Zasady postepowania diagnostycznego.

The paper presents a detailed analysis of errors in the diagnosis of bacteriologically negative pulmonary tuberculosis in a random sample of 560 patients out of all such patients registered in 1993. The false diagnosis was found in 63 patients i.e. in 11.3% of the sample. Among the 63 patients with false diagnosis of tuberculosis there were 15 cases with lung cancer, 15 cases with pneumonia and/or pleuritis, 15 cases with disease of circulatory system with abnormal radiological of the lungs, 8 cases with old post-tuberculosis changes in the lung considered as relapses, 4 cases of bronchiolitis obliterans with organizing pneumonia, 3 cases of sarcoidosis and 1 case of aspergillosis in a post-tb cavity. The most serious errors were those related to failure in diagnosing (or of too late diagnosis) of lung cancer. These failures comprised 2.7% of patients in the sample. The chance to detect a lung cancer in the sample was 2900 cases in 100,000 men and 2200 cases in 100,000 women. This chance was much higher than in general population--29x for men and 100x for women. The main source of errors was false interpretation of radiological examinations and neglect to utilise other diagnostic procedures like bronchoscopy, cytology or tomography examinations. As an outcome of the analysis authors present their recommendations for diagnostic procedures in the diagnosis of bacteriologically negative pulmonary tuberculosis.

[Analysis of patients registered in outpatient departments for tuberculosis and lung diseases in group VI N (neoplastic) during the first half of 1995]. / Analiza chorych zarejestrowanych w poradniach gruzlicy i chorób pluc w grupie VI n (nowotworowej) w pierwszym Pólroczu 1995 r.

2115 cases of chest neoplasms were registered during the first six months of 1995 in Pulmonary Outpatient Departments in Poland. In 865 (40.8%) patients squamous cell cancer was diagnosed, in 344 (16.2%)--small cell lung cancer, in 174 (8.2%)--adenocarcinoma, in 107 (5%) other types of lung cancer and in 5.6% of cases--lung cancer with undefined histology. Predominated subjects aged from 60 to 69 years (43.5%). Lung cancer under 50 years of age was observed significantly more often among women (25.8%) than among men (13.5%). Moreover adenocarcinoma was much more frequent among women than men and significantly more cases of lung cancer were recorded in nonsmoking women. Diagnosis was established during the first 4 weeks in 38.9% of patients but 23.6% of patients have waited for it more than 39 weeks. Surgical treatment was advised in 16.9% of subjects, radiotherapy in 14.3% and 26.4% of patients underwent chemotherapy. Treatment analysis was presented according histological type of lung cancer. Occupation, performance status, clinical stage of the disease, prevalence of cancer in patients families were also analysed.

[Chemotherapy versus chemoradiotherapy in patients with limited small cell lung carcinoma]. / Chemioterapia versus naprzemienna chemio-radioterapia u chorych z ograniczona postacia drobnokomórkowego raka pluca.

62 patients with a limited small cell lung cancer were randomly qualified into two groups. 32 patients of the first group were treated only with the chemotherapy regimen, consisted of three drugs (Carboplatine, Etoposide and Vincristine administered in 6 courses, on regular, 3-weeks basis). The second group of 30 patients had been treated with the identical chemotherapy schedule, but alternatively combined with a primary site irradiation in a total dose of 40Gy, applied in parts after the chemotherapy courses 2, 3, and 4. The significantly higher proportion of a complete remission results was observed in the alternate-treatment group: 14/30 (46.7%), compared with the chemotherapy-only group: 10/32 (31%). Alternate chemoradiotherapy resulted both in the increased median remission duration time, and the increased median survival time. Only in the alternate chemotherapy group, in 14/30 patients (46.7%) the pneumotoxicity symptoms appeared, whilst no differences in other organ-specific treatment-induced toxic effects were noted.

[Comparison of conventional chest radiography and computed tomography for evaluation of the degree of regression in neoplasms of patients treated for small cell lung cancer]. / Porównanie konwencjonalnego badania radiologicznego klatki piersiowej i tomografii komputerowej w ocenie stopnia regresji zmian nowotworowych u chorych leczonych z powodu drobnokomórkowego raka pluca.

The aim of the study was to assess how objective is the estimation of the chest lesions regression on RT picture and how extent chest CT can increase the precision of this assessment. The material consisted of 66 SCLC pts observed in the Institute of Tuberculosis and Lung Diseases in Warsaw between 1987 and 1994 in whom RT and CT were performed to estimate the degree of response to treatment. Chest RT were estimated by 4 specialists independently. In 28 cases (42%) opinion concerning the degree of tumor regression was the same. In 23 cases (35%) the opinions were discordant and in 15 cases (13%) (mainly after chest irradiation) all examiners agreed that degree of response is impossible to precise (it ranged between PR and CR). In the second part of this study the degree of cancer regression was assessed in the same patients by 2 specialists independently estimated CT pictures. Opinions were different only in 4 cases. The comparison between these two methods was done. CI examination allowed exact estimation of response degree in 20 out of 24 pts (83%) in whom this assessment was impossible using chest radiography. In 9 cases (21%) the assessment of chest CT changed the previous estimation made using chest radiography (in 5 cases from CR to PR and in 4 cases from PR to CR). In conclusion-estimation of tumor regression using chest RT is very subjective method and many errors are possible. Chest CT estimation is a much better method for estimation of tumor response especially in irradiated patients.

[Diffuse alveolar hemorrhage in Wegener's granulomatosis]. / Rozlane krwawienia pecherzykowe w przebiegu ziarniniakowatosci Wegenera.

Two cases of Wegeners Granulomatosis were presented. In the both cases diffuse alveolar hemorrhage was found.

The role of human chorionic gonadotropin beta subunit elevation in small-cell lung cancer patients.

Human chorionic gonadotropin (HCG)-like immunoreactivity has been found in many non-trophoblastic tumours, but the biological behaviour of HCG-producing cells has not been clarified yet. The aim of the study was to estimate the frequency of serum HCG beta subunit (s beta HCG) elevation in patients with small-cell lung cancer (SCLC) and to assess its possible prognostic role in this type of tumour. An attempt was also made to reclassify the histology in selected cases to see whether the elevated (s beta HCG) level is connected with any special subtype of small-cell lung cancer. A total of 156 SCLC patients entered the study: 93 men, 63 women, median age 58 years. s beta HCG activity was measured by immunoenzyme assay (Abbott EIA beta HCG 15-15) before treatment. s beta HCG elevation (above 5 mIU/ml) was found in 21 of 156 patients (14%). Response to treatment after chemotherapy (complete and partial response) was obtained in only 48% of those patients in whom elevated s beta HCG was found, in comparison to the 73% response rate observed in the remaining patients. Only 5% of patients with elevated s beta HCG survived 2 years, in comparison to 21% surviving for 2 years among the remaining patients. The prognostic significance of elevated s beta HCG and extent of disease were independent of each other (Cox's proportional-hazard model). Thus s beta HCG elevation in SCLC seems to be a marker of more resistant tumours and of poor prognosis. We have not found any connection between the subtype of small-cell lung cancer and elevated s beta HCG. Elevated s beta HCG was found in 2 out of 11 patients with oat-cell carcinoma, in 3 out of 10 patients with an intermediate cell type and in 5 out of 13 patients with small-cell lung cancer in which the assessment of the subtype was not possible.

Subacute sensory neuronopathy in small cell cancer of the lung. Immunocytochemical study of 2 cases.

Two cases of subacute sensory neuronopathy (SSN) associated with small cell lung cancer are reported. In both cases motor disability, attributed to motor neuropathy and neuronopathy, respectively, accompanied SSN. Immunohistochemical studies performed provide further data suggesting the involvement of humoral antibodies and the participation of protease inhibitors in the pathogenesis of SSN.

Effect of purinergic receptor antagonists suramin and theobromine on tumor-induced angiogenesis in BALB/c mice.

The purinergic receptor antagonists suramin (SRN) and theobromine (TBR) were examined for their anti-angiogenic activity in BALB/c mice. SRN or TBR were subcutaneously administered to BALB/c mice in doses of 1-125 mg/kg body weight on days 0, 1 and 2 after intradermal inoculation of E14/W lung carcinoma cells. It was shown that SRN and TBR inhibited tumor-related angiogenesis. Accordingly, it may be suggested that purinoceptor antagonists may inhibit neovascularization in tumor growth and metastasis.

[Angiogenesis in the neoplastic process]. / Angiogeneza w procesie nowotworowym.

Solid tumor growth is accompanied by neovascularisation. In the absence of neovascularisation most tumors might become dormant at a tiny diameter 2-3 mm. New capillary growth is elicited by a diffusible factors generated by malignant tumor cells and by host cells. The inhibition of angiogenesis may provide a form of cancer treatment, either substitutional or additional to the conventional form of therapy. The present review summarized the state of our knowledge on this topic, angiogenesis and tumor growth.