Multielemental Analysis of Tobacco Plant and Tobacco Products by TXRF.

The aim of this research is to develop a fast analytical method for multielemental analysis of the tobacco plant Virginia tobacco (cultivated in Poland) and tobacco products (cigarettes, cigars, cigarillos, snuff and two kinds of properly crafted tobacco such as a shisha and cigarette tobacco) distributed in Polish markets by means of a low-power benchtop total reflection X-ray fluorescence (TXRF) system. For this purpose, a set of certified tobacco materials and real samples was employed. In leaves and stalks of V. tobacco and tobacco products, a concentration of 18 elements (P, S, Cl, K, Ca, Ti, Cr, Mn, Fe, Ni, Cu, Zn, As, Se, Br, Rb, Sr and Pb) was determined. Analyzing elemental composition of tobacco plants, one can see that concentrations of S, Ca, Ti, Mn, Zn, Sr and Pb are higher in leaves, whereas the concentrations of P, Cl, K, Fe Cu and Br are higher in stalks; the levels of Cr, Ni, As and Rb are comparable in both these parts of the tobacco plant. All of the parameters affecting sample preparation and TXRF measurements conditions were carefully evaluated. The accuracy and precision of the TXRF measurements were verified using an internal standardization approach for quantification.

CHEK2 variants predispose to benign, borderline and low-grade invasive ovarian tumors.

OBJECTIVE: Three founder alleles of the CHEK2 gene have been associated with predisposition to a range of cancer types in Poland. Two founder alleles (1100delC and IVS2 + 1G >A) result in a truncated CHEK2 protein and the other is a missense substitution, leading to the replacement of a threonine with an isoleucine (I157T). METHODS: To establish if these variants play a role in the etiology of ovarian tumors, we genotyped 1108 Polish women with various types of ovarian tumors and 4000 controls for the three CHEK2 variants. We included 539 Polish women with benign ovarian cystadenomas, 122 women with borderline ovarian malignancies and 447 women with invasive ovarian cancer. RESULTS: Positive associations were seen with the CHEK2 I157T missense variant and ovarian cystadenomas (OR = 1.7; P = 0.005), with borderline ovarian cancers (OR = 2.6; P = 0.002) and with low-grade invasive cancers (OR = 2.1; P = 0.04). There was no association with ovarian cancer of high grade (OR = 1.0). The association between the I157T missense variant was then confirmed in a second sample of Russian patients with borderline ovarian cancers (OR = 2.7; P = 0.06). CONCLUSION: These data indicate that CHEK2 variants may predispose to a range of ovarian tumor types of low malignant potential, but not to aggressive cancers.

Prognostic value of S-100 immunostaining in tumour cells of non-small cell lung cancer.

S-100 protein expression is present in various malignant tissues, yet its prognostic relevance is debatable. The aim was to assess in non-small cell lung cancer (NSCLC) patients' prognostic value of S-100 protein considered alone or in relation with other variables. Tumour samples taken from 86 NSCLC patients during resection were assayed for S-100 protein expression with the use of polyclonal DAKO ZO311 antibody. S-100 expression was found in 32 cases (37%). Positive staining was not correlated with clinical characteristics including age, sex, pathology type of tumour, stage and cigarette smoking. There was a tendency for simultaneous expression of S-100 and P53 protein (p=0.06). A median survival rate for the entire group was 2.3 years (95% CI, 0.9-3.6 years). The median and 5-year survival of patients with positive staining for S-100 protein was 1.5 years and 25%, respectively, compared with 3.0 years and 35%, respectively, in the S-100 negative group (p=0.17). In the final model of a multivariate analysis, S-100 protein expression in tumour cells was associated with significantly decreased survival (p=0.005). S-100 protein expression in tumour cells seems to be an independent predictor of poor prognosis in NSCLC patients.

BRCA1-positive breast cancers in young women from Poland.

We identified 4316 unselected incident cases of early-onset breast cancers (<51 ears of age at diagnosis) in 18 Polish hospitals between 1996 and 2003. We were able to obtain a blood sample for DNA analysis from 3472 of these (80.4%). All cases were tested for the presence of three founder mutations in BRCA1. The proportion of cases with a BRCA1 mutation was 5.7%. The hereditary proportions were higher than this for women with breast cancer diagnosed before age 40 (9%), for women with cancer of medullary or atypical medullary histology (28%), for those with bilateral cancer (29%) or with a family history of breast or ovarian cancer (13%). It is reasonable to offer genetic testing to women with early-onset breast cancer in Poland.

Germline MSH2 and MLH1 mutational spectrum including large rearrangements in HNPCC families from Poland (update study).

Germline mutations in the DNA mismatch repair genes MSH2 and MLH1 account for a significant proportion of hereditary non-polyposis colorectal cancer (HNPCC) families. One approach by which development of an efficient DNA-testing procedure can be implemented is to describe the nature and frequency of common mutations in particular ethnic groups. Two hundred and twenty-six patients from families matching the Amsterdam II diagnostic criteria or suspected HNPCC criteria were screened for MSH2 and MLH1 germline mutations. Fifty different pathogenic mutations were found, 25 in MSH2 and 25 in MLH1. Twenty-four of these had not previously been described in other populations. Among our 78 families with MSH2 or MLH1 mutations, 54 (69.2%) were affected by recurrent mutations including 38 found at least twice in our own series. Two of the most frequent alterations were a substitution of A to T at the splice donor site of intron 5 of MSH2 and a missense change (A681T) of MLH1 found in 10 and eight families, respectively. Among large deletions detected by the multiplex ligation-dependent probe amplification assay, exon 9 deletions in the MSH2 gene were found in two families. Our results indicate that a screening protocol specific for the Polish population that is limited to the detection of all reported mutations will result in the identification of the majority of changes present in MLH1 and MSH2 genes in Polish HNPCC kindreds.

The repair of gamma-radiation-induced DNA damage is inhibited by microcystin-LR, the PP1 and PP2A phosphatase inhibitor.

The genotoxic activity of microcystin-LR (MC-LR) is a matter of debate. MC-LR is known to be a phosphatase inhibitor and it may be expected that it is involved in the regulation of the activity of DNA-dependent protein kinase (DNA-PK), the key enzyme involved in the repair of radiation-induced DNA damage. We studied the effect of MC-LR on the repair capacity of radiation-induced DNA damage in human lymphocytes and human glioblastoma cell lines MO59J and MO59K. A dose of 0.5 microg/ml of MC-LR was chosen because it induced very little early apoptosis which gives no false positive results in the comet assay. Human lymphocytes in G0-phase of the cell cycle were pre-treated with MC-LR for 3 h and irradiated with 2 Gy of gamma radiation. The kinetics of DNA repair was assessed by the comet assay. In addition the frequencies of chromosomal aberrations were analysed. The pre-treatment with MC-LR inhibited the repair of radiation-induced damage and lead to enhanced frequencies of chromosomal aberrations including dicentric chromosomes. The results of a split-dose experiment, where cells were exposed to two 1.5 Gy doses of radiation separated by 3 h with or without MC-LR, confirmed that the toxin increased the frequency of dicentric chromosomes. We also determined the effect of MC-LR and ionizing radiation on the frequency of gamma-H2AX foci. The pre-treatment with MC-LR resulted in reduced numbers of gamma-H2AX foci in irradiated cells. In order to elucidate the impact of MC-LR on DNA-PK we examined the kinetics of DNA repair in human glioblastoma MO59J and MO59K cells. Both cell lines were exposed to 10 Gy of X-rays and DNA repair was analysed by the comet assay. A strong inhibitory effect was observed in the MO59K but not in the MO59J cells. These results indicate that DNA-PK might be involved in DNA repair inhibition by MC-LR.

A common variant of CDKN2A (p16) predisposes to breast cancer.

BACKGROUND: A common missense variant of the CDKN2A gene (A148T) predisposes to malignant melanoma in Poland. An association between malignant melanoma and breast cancer has been reported in several families with CDKN2A mutations, OBJECTIVE: To determine whether this variant also predisposes to breast cancer. METHODS: Genotyping was undertaken in 4209 cases of breast cancer, unselected for family history, from 18 hospitals throughout Poland and in 3000 controls. RESULTS: The odds ratio (OR) associated with the CDKN2A allele for women diagnosed with breast cancer before the age of 50 was 1.5 (p = 0.002) and after age 50 it was 1.3 (p = 0.2). The effect was particularly strong for patients diagnosed at or before the age of 30 (OR = 3.8; p = 0.0002). CONCLUSIONS: CDKN2A appears to be a low penetrance breast cancer susceptibility gene in Poland. The association should be confirmed in other populations.

Copper, zinc, and selenium in whole blood and thyroid tissue of people with various thyroid diseases.

We investigated the possible differences among the concentrations of copper, zinc, and selenium, and their mutual relations in the whole blood and thyroid tissue of patients with various thyroid disorders. Trace elements were determined by total-reflection X-ray fluorescence. The mean levels of these metals in blood as well as the mean Cu/Zn, Cu/Se, and Zn/Se ratios in the patients with thyroid cancer were significantly higher that in other patients and the control groups. However, the mean Zn and Se concentrations in the thyroid cancer tissue were significantly lower than in the thyroid tissue of other patients. In addition, the mean Cu/Zn and Cu/Se ratios in the thyroid cancer tissue were significantly higher than in the patients with other thyroid diseases. We confirm that the highest levels of copper and zinc as well as the Cu/Zn, Cu/Se, and Zn/Se ratios in the whole blood of the patients with thyroid cancer may suggest the progression of the proliferation process in the thyroid gland. We suggest that the low concentrations of zinc and selenium in the thyroid tissue confirm their participation in the carcinogenic process.

Concentration of selenium in the whole blood and the thyroid tissue of patients with various thyroid diseases.

We investigated the possible differences between the concentrations of selenium in the whole blood and thyroid tissue of patients with thyroid disease. The study comprises 41 women with nodular goiter, 19 women and 2 men with thyroid cancer, 18 women with Graves' disease, and 7 women with thyroiditis. The concentration of selenium was determined by the TRXRF method. The lowest mean selenium level was achieved in the whole blood of women with Graves' disease and the highest in the whole blood of healthy people. In the thyroid cancer tissue, we found the lowest concentration of selenium and the highest in the thyroid gland of women with nodular goiter and Graves' disease. The low selenium levels in the thyroid tissue may increase thyroid cancer risk.

A multicentre randomized/controlled trial of a conventional versus modestly accelerated radiotherapy in the laryngeal cancer: influence of a 1 week shortening overall time.

BACKGROUND AND PURPOSE: To compare in a phase III study the loco-regional control, disease-free survival and overall survival induced by an accelerated regimen (AF) as compared with conventional regimen (CF) and to analyze the early and late post-radiation morbidity in both arms. MATERIALS AND METHODS: Patients with age < or = 75, WHO 0-1, suitable for a radical course of radiotherapy T1-T3, N0, M0, stage of glottic and supraglottic laryngeal cancer were randomized to either CF: 66Gy given in 33 fractions over 45 days or AF: 66Gy given in 33 fractions over 38 days (2 fractions every Thursday). A total of 395 patients were included from 05.1995 to 12.1998. RESULTS: Early toxicity: At the end of radiotherapy patients treated with AF complained for more severe reactions than patients treated with CF. In 8 weeks after treatment completion patients treated with AF complained only for more severe pain on swallowing (P=0.027). In 4 months after treatment completion all types of toxicity except for skin teleangiectasia (P=0.001) were similar in the two groups. Loco-regional control: comparison between CF and AF showed no difference in terms of loco-regional control (P=0.37). CONCLUSIONS: The improvement in AF in terms of loco-regional control is estimated to be 3-5% in comparison with conventional regimen and is not significant. The intensity of reactions after 4 months was similar in both arms, what suggests the possibility of further shortening of the overall time by few days or enhancing the total dose within the limits of acceptable morbidity.

Prognostic relevance of proliferating cell nuclear antigen and p53 expression in non-small cell lung cancer.

Prognostic value of p53 and PCNA expression in non-small cell lung cancer (NSCLC) remains controversial. In this study we determined the relevance of these abnormalities in terms of overall survival and disease-free survival in 95 NSCLC patients who underwent curative pulmonary resection. Expression of p53 was found in 44 samples (45%), expression of PCNA-in 79 samples (83%), and expression of both markers-in 35 samples (36%). There was no relationship between expression of either protein and major clinicopathological characteristics. Median survival for patients with and without p53 expression was 36 and 33 months, respectively and 5-year survival probability-29 and 37%, respectively (P=0.73). Median survival for patients with and without PCNA expression was 36 and 27 months, respectively and 5-year survival probability-35 and 25%, respectively (P=0.60). There was no significant difference in overall survival between particular groups of patients with tumors carrying four possible p53/PCNA phenotypes. In multivariate analysis including patient age, sex, tumor stage, tumor type and differentiation, p53 and PCNA expression, the only variable important for survival was stage of disease. These results suggest the lack of prognostic relevance of p53 and PCNA expression in surgically treated NSCLC patients.

Zn concentration in thyroid tissue and whole blood of women with different diseases of thyroid.

The Zn concentration in thyroid tissue and whole blood of patients with Graves' disease, thyroid cancer, and nodular goiter disease was determined using the total-reflection X-ray fluorescence method. The dependence of obtained concentrations on the clinical stage of the examined disease, histopathological grading, and kind of analyzed material (thyroid tissue and blood) was studied. The determined concentration of Zn was the lowest in the thyroid tissue of patients with thyroid cancer (23.1 microg/g) and it was the highest in the case of Graves' disease (41.7 microg/g), whereas in the blood samples, the reverse results were found (7.1 microg/g and 4.8 microg/g, respectively). The physical basis of the method used, the experimental setup, and the procedure of sample preparation are described.

Humoral immune response after vaccination against influenza in patients with breast cancer.

The aim of this pilot study was to assess a humoral response to influenza vaccine in 9 women with breast cancer. In the epidemic season 1998/1999 the patients received a single 0.5-ml dose of split influenza vaccine (Vaxigrip, Pasteur Merieux). Humoral response was measured by the hemagglutinin inhibition test in sera collected before vaccination and 1 month after vaccination. All results were compared with a control group of 19 healthy vaccinated women. The mean 'fold' increases ranged from 12.0 to 22.2 in patients with breast cancer and from 10.5 to 29.2 in healthy women. After vaccination, protection rates ranged from 44.4% to 88.9% and 63.2% to 94.7%, respectively. Response rates were between 44.4% and 88.9% in women with cancer and between 63.2% and 78.9% in the control group. After vaccination, antihaemagglutinin antibody titers were significantly higher than the prevaccination titers. During the whole study there were no statistically significant differences in humoral response between patients with breast cancer and healthy women. The results of the present study clearly show that women with breast cancer, including those undergoing chemotherapy, were able to develop a good serological response to influenza vaccine.

[Clinical-pathomorphologic preparation of patients after surgical treatment of differentiated thyroid cancer to adjuvant treatment with 131I]. / Kliniczno-patomorfologiczne przygotowanie chorych po chirurgicznym leczeniu zróznicowanych raków tarczycy do uzupelniajacego leczenia 131I.

UNLABELLED: Thyroid cancer treatment includes: surgery, radioiodine therapy, thyroxine therapy and radiotherapy. Selection of treatment's strategy depends on histopathological evaluation, age of patient and iodine uptake. The aim of the analysis was to verify how the medical documentation of patients treated by surgical treatment was prepared. All patients were directed to the Department of Endocrinology, Holycross Cancer Center in Kielce. Analysis encompassed 33 patients classified to 131I therapy for the first time. In each case patient's medical documentation and histopathological diagnosis made in Department of Tumor Pathology, Holycross Cancer Center, were compared. RESULTS: A conformity with primary cancer diagnosis was obtained in all but one patient in whom pathological consultation did not confirm oxyphilic carcinoma. In four cases the type of cancer was changed as a result of repeated consultation. pTMN classification was stated in 23 primary pathologic examination cases. After consultation of slides and inspection of surgery protocol, total or partial pTNM stage was obtained in other 9 patients. CONCLUSIONS: Establishment of diagnostic and therapeutic algorithm, accepted on Conference in Szczyrk, 1995 is inadequately executed in small centers. Routine consultation of histopathological slides creates a possibility to make a proper choice of treatment's strategy.

[Prognostic value of P53 protein in cells of non-small cell lung cancer]. / Ocena rokowniczego znaczenia obecnosci bialka P53 w komórkach niedrobnokomórowego raka pluca.

The aim of the study was to evaluate the prognostic relevance of p53 protein in non-small cell lung cancer. The 95 surgically treated patients were included (53 patients with squamous cell carcinoma, 29--with adenocarcinoma, 5--with large cell carcinoma, and 8--with mixed type). The protein was assessed immunohistochemically with the use of monoclonal antibodies DO7, DAKO. Positive staining was present in 44 patients. There was no survival difference between groups with and without protein (median survival--36 and 33 months, respectively; p = 0.86). In the multivariate analysis the only characteristics with prognostic impact in the entire group was stage of the disease. There was no correlation between the expression of p53 protein and disease-free survival. These results indicate that there is no prognostic relevance of p53 protein in non-small cell lung cancer.

[Histopathological verification of the cervical pap smears based on material of the obstetrical-gynecological department of the provincial hospital in Kielce]. / Histopatologiczna weryfikacja rozmazów cytologicznych szyjki macicy.

The results of the cervical Pap. smear taken before cold knife conization and the histopathological results of the cones removed in 336 women, were compared. In cytological examination we found 10.8% (13/120) false negative results, which did not confirm of the existence the precancerous lesion within cervical epithelium. The mild dysplasia according to Pap. smears was established among 15.9% (15/94) of women with CIN III and CIN II diagnosed pathologically. In part of the women the histopathological results of the postoperative specimens and bioptic cervical material were compared. Clinical value and correlation between histopathological and cytological results in the estimation of CIN (cervical intraepithelial neoplasia) were discussed with particular regard to CIN I according to Bethesda system.

An elemental correlation study in cancerous breast tissue by total reflection x-ray fluorescence.

The total reflection x-ray fluorescence method (TRXRF) has been employed to determine of P, S, K, Ca, Cr, Mn, Fe, Ni, Cu, Zn, Se, Rb, Sr, and Pb concentration in the benign breast tumor tissue from 68 women and in the cancerous breast tissue from 26 women. Concentrations of most of elements show enhancement in cancerous breast tissue. Examined elements compete for binding sites in the cell, change its enzymatic activity, and exert direct or indirect action on the carcinogenic process accelerating the growth of tumors. Inhibition of enzymatic activity caused by variation in trace element concentrations results in immunological breakdown of the body system. An attempt has been made to correlate measured trace element concentrations with the clinical stage of cancer. Physical bases of used analytical method, experimental setup, and the procedure of sample preparation are described.

[CA 19-9 and CA 125 antigens in the sera of patients with cancer of the large intestine in relation to its clinical progress]. / Stezenie antygenu CA 19-9 i CA 125 w surowicy krwi chorych na raka jelita grubego w zaleznosci od zaawansowania klinicznego.

By radioimmunoassay the concentration of the CA 19-9 antigen was determined in the serum of 68 patients with large bowel cancer, while the CA 125 antigen was determined in 26 patients with this disease. Both markers were determined with CIS kits. In all, 127 determinations were done. The results were as follows: 1) CA 19-9 concentration increased with disease progression from 21% (in grade I of local-regional progression, A, B, C according to Dukes), through 41% above-normal results in group II (with metastases--Dukes' grade D), to 67% in case of recurrence of the tumor, and to 0% in group IV--with absent recurrence sign after radical surgical intervention. 2) CA 125 is without clinical value in large bowel cancer.

Pretreatment concentrations of breast carcinoma antigen (CA 15.3) and mucin-like carcinoma-associated antigen in patients with carcinoma of the breast.

Pretreatment serum concentrations of breast carcinoma antigen (CA 15.3) and mucin-like carcinoma-associated antigen (MCA) were determined in 129 patients with breast carcinoma. Concentrations of both markers were within the normal range in patients with Stage I disease. Concentrations of CA 15.3 were elevated (greater than 40 U/ml) in 3, 11 and 48%, those of MCA (greater than 17 U/ml) in 11, 18 and 52%, and those of one or the other marker in 11, 18 and 58% of the patients with Stage II, III and IV disease, respectively. The elevation of either marker roughly paralleled the size of the tumor being normal in the patients with localized cancer, slightly elevated in a small proportion of the patients with locoregional cancer, and moderately to markedly elevated in half of the patients with distant metastases. Correlation between serum concentrations of CA 15.3 and MCA was highly significant (p less than 0.0001). It is concluded that the markers were equally sensitive and that an elevated serum level was a useful adjunct for staging, implying systemic disease.