Variations in steroid hormone receptor content throughout age and menopausal periods, and menstrual cycle in breast cancer patients.

Variations in steroid hormone receptor contents throughout age and menopausal periods define three breast carcinoma groups: younger premenopausal carcinomas (aged up to 45), middle-aged carcinomas (pre-, peri- and postmenopausal aged 45-59) and older postmenopausal carcinomas (aged over 59). Age-related steroid hormone receptor contents within premenopausal and postmenopausal carcinoma groups are characterized by the important increase of both receptor contents, while menopausal-related steroid hormone receptor contents within middle-aged carcinoma group (aged 45-59) are characterized by the important decrease of progesterone receptor content and estrogen receptor functionality. No variations in steroid hormone receptor contents throughout menstrual cycle within the follicular and the luteal phases were obtained. The important decrease of estrogen receptor content in the mid-cycle phase versus the perimenstrual phase was found. Variations in steroid hormone receptor contents throughout age and menopausal periods, as well as throughout menstrual cycle could not be associated with variations in the blood steroid hormone concentrations. However, important association between steroid hormone receptor contents and the blood steroid hormone concentrations was found within the luteal phase carcinoma group and within older postmenopausal carcinoma group. It is interesting that within carcinoma group with the highest concentration of progesterone, progesterone receptor content increases with an increase of the ratio of estradiol and progesterone blood concentrations, while within carcinoma group with the lowest steroid hormone concentration and the highest content of estrogen receptor content, estrogen receptor content decreases with an increase of either the blood estradiol concentration or the ratio of the blood estradiol and progesterone blood concentrations.

Lysosomal dipeptidyl-peptidases I and II in human squamous cell lung carcinoma and lung parenchyma.

In the present work we studied the levels of activities of dipeptidyl-peptidase I (or cathepsin C, DPP-I) and dipeptidyl-peptidase II (DPP-II) and examined their isoelectric focusing profiles in matched pairs of human squamous cell lung carcinoma (SQCLC) and the lung from surgically treated patients (n = 33). The mean specific activities of DPP-I and DPP-II were higher in SQCLC (Stages I and II) than in the lung, but only the activity of DPP-II in Stage I SQCLC was significantly higher compared to the lung. The activities of both enzymes were higher in the tumor than in the lung in 10 of 20 Stage I SQCLC patients, but only in 3 of 13 Stage II SQCLC patients. The specific activities of DPP-I and DPP-II in the lungs showed a good correlation while the correlation of both enzyme activities in SQCLCs was poor. We observed only a small and mutually comparable activation of DPP-I in extracts from SQCLCs and from the lungs by dithiothreitol. The isoelectric focusing profile of several DPP-II forms in SQCLCs and the lungs was similar and the single major DPP-II isoform revealed in the tumors and lungs showed a pIapp of 5.3-5.2. The isoelectric focusing profile of DPP-I showed multiple enzyme forms in SQCLCs (pIapp 6.3-4.5) as well as in the lungs (pIapp 6.4-4.8). In SQCLCs, as well as in the lungs, the activities of the DPP-I forms with pIapp values < or = 5.6 were shifted by neuraminidase treatment to the site of the major DPP-I isoform with pIapp of about 6.0 and the zymograms then showed an another DPP-I with pIapp of 5.7, which was less discernible in the lung. In some patients, the DPP-I forms with pIapp values < or = 5.6 from SQCLC retained a greater percentage of activity distribution than did the DPP-I pIapp-counterparts from the lung.

Effects of sequence and temperature of hyperthermia and peplomycin on human pharyngeal carcinoma KB cells in vitro.

To maximize the interactive effect, we examined the time sequence of high (above 43 degrees C) or low (below 43 degrees C)-hyperthermia and peplomycin (PEP)(0.5 microgram/ml). Simultaneous or post-hyperthermic PEP treatment at 44 degrees C resulted in a slight synergistic effect with thermoenhancement ratios (TER) of 1.30 or 1.34, respectively. However, at 42 degrees C, maximal interaction was obtained (TER = 10.19) when KB cells were simultaneously heated with PEP. Furthermore, pre-treatment at 44 degrees C for 25 min or 42 degrees C for 4 h enhanced PEP cytotoxicity more than that of post-treatment at 44 degrees C for 25 min or 42 degrees C for 4 h, respectively. However, chemoenhancement ratios (CER) of pre-treatment at 44 degrees C for 25 min and at 42 degrees C for 4 h were 19.1 and 3.5, respectively, although the isothermic dose decreased the cell count to 60% in both cases. These results indicated that simultaneous or post-hyperthermic PEP treatment with high-hyperthermia, and simultaneous PEP treatment with low-hyperthermia, are the most effective means of PEP thermochemotherapy with hyperthermia.

Detection of human papillomavirus (HPV) type 6, 16 and 18 in head and neck squamous cell carcinomas by in situ hybridization.

Seventy seven squamous cell carcinomas (10 oral cavity, 15 tongue, 26 pharynx and 26 larynx), with different grading, were analyzed for the presence of HPV DNA by in situ hybridization. Positive signals were found on the nuclei of cancer cells in 25 (32.5%), in the epithelia adjacent to squamous cell carcinomas in 2 (8.7%), and in the resected margins in 1 (4.3%) case. HPV DNA positive signals were obtained in 42% of laryngeal, 34% of pharyngeal, in 20% of oral, and 20% of tongue carcinomas. Out of 25 HPV positive carcinomas a single HPV type was detected in at least 11 (44%), and double or multiple infection in 9 (36%) cases; altogether, HPV 6 DNA was determined in 15 (60%), and HPV 16 and/or 18 DNA in 17 (68%) head and neck tumors. The detection rate of HPV 6 was lower than of HPV 16 and/or 18 for tumors in oral cavity, tongue and larynx. Out of 25 HPV DNA positive carcinomas 21% were graded as G1, 27% as G2, and 44% were G3. The results indicate that HPV may be involved in the pathogenesis of head and neck squamous cell carcinomas.

The value of prognostic factors in the management of stage I nonseminomatous germ cell testicular tumors (NSGCTT).

The prospective study, carried out from February 1992 to January 1996, included 49 patients in clinical Stage I nonseminomatous germ cell testicular tumors (NSGCTT). They were aged 16-40 years (mean, 25 years). Patients were stratified to different risk-adapted therapeutic approaches according to histopathologic findings of primary tumor removed by inguinal orchiectomy. Eleven patients of the 1st group with vascular invasion and majority of embryonal carcinoma components in the primary tumor were treated with adjuvant chemotherapy (2 cycles of BEP). None of them had disease progression after the follow-up of 4-43+ months (mean, 20.9 months) after orchiectomy. Five patients of the 2nd group with vascular invasion and majority of teratoma elements in the primary tumor were treated with primary retroperitoneal lymph node dissection (RPLND). They were followed-up 29-45+ months (mean, 33.4 months) after orchiectomy. Two of them (40%) had pathologic Stage II after RPLND and underwent subsequent BEP chemotherapy. One of them died due to disease progression in disseminated stage 29 months after orchiectomy. The second one lives with no evidence of the disease (NED). Thirty three patients in the 3rd group without vascular invasion were kept under surveillance. They were followed-up 3-48+ months (mean, 22.3 months) after orchiectomy. Disease progression was observed in 5 of them (15.1%), 7-10 months (mean, 8.8 months) following orchiectomy. These patients were treated with BEP chemotherapy and live with NED 1-16+ months (mean, 9.2 months) after completion of the therapy. The overall survival rate in clinical Stage 1 patients was 97.9%. The authors recommend the surveillance policy only in clinical Stage I NSGCTT patients without vascular invasion in the primary tumor.

Gastric cancer in the province of Zaragoza (Spain): a survival study.

A study of the main prognostic factors and of the overall survival rate of gastric cancer (GC) is presented. It covered 895 cases diagnosed histopathologically in the province of Zaragoza (Spain) over a ten-year period (1980-1989). The analysis of the survival rate was carried out according to the Kaplan-Meier method and the Mantel-Haenszel test. The average overall survival rate of the sample was 6.5 months and the five-year survival rate was 16.5%. Lauren's intestinal histological type is associated with a better prognosis (a five-year survival rate of 25%) than the diffuse type (15%). The survival rate with regard to gastric wall invasion ranges from 78% for T1 tumors to 8% for T4 tumors (p < 0.0001). There are significant differences in survival rate between the TNM classification stages, ranging from a five-year survival rate of 77% for Stage I to 0% for Stage IV.

Activity and characterization of sialyltransferase from serum of normal rats, of rats bearing Zajdela ascitic hepatoma, in normal host liver and in Zajdela hepatoma cells.

Comparative studies on the content of sialic acid and on the sialyltransferase activity in normal serum and in serum of rats with Zajdela ascitic hepatoma in different phases of tumor development have been conducted. Unlike the serum from animals with tumors, in which the sialic acid quantity increases in dependence of the stage of tumor development, the activity of serum sialyltransferase statistically augmented only in serum of rats at the final stage of tumor progression. The sialyltransferase activity towards asialofetuin as an acceptor in normal liver and in Zajdela hepatoma cells, was measured and a decrease in this activity in tumor cells as well as in host liver was found. When lactose was used as acceptor, again lower enzyme activity in the tumor cells in comparison with that in liver was established, but in liver and in hepatoma cells the predominant 14C-labelled product of the sialyltransferase assay was alpha (2-6) sialyllactose isomer. The results contribute to the biochemical characterization of rat Zajdela hepatoma.

Evaluation of phthalmustine, a new anticancer compound. II. Tumor inhibitory effect in murine Ehrlich ascites carcinoma and sarcoma-180.

The antitumor activity of phthalmustine (Neoplasma, 41, 1994, 35) has been evaluated in murine Ehrlich ascites carcinoma (EAC) and sarcoma-180 (S-180). The hematological changes associated with the application of phthalmustine were also evaluated in tumor bearing as well as in normal male Swiss mice. The results indicate that phthalmustine induces marked inhibition of tumor growth and substantially prolongs host survival having curative effect while it does not adversely affect hematopoiesis at the optimum dose tested. No toxic symptoms were noted with respect to external appearance, body weight changes and behavioral pattern.

Protein kinase inhibitors: induction of apoptosis and drug-resistance modulation in neoplastic cell lines.

Overcoming of P-glycoprotein mediated vincristine resistance of L1210/VCR mouse leukemic cells could be induced by pentoxifyline but not by theophylline and caffeine.

Effects of xanthine derivatives (pentoxifylline, caffeine, theophylline, 1-methyl-3-isobutylxanthine) on P-glycoprotein mediated vincristine resistance of L1210/VCR mouse leukemic cell subline were studied. From the applied xanthines only PTX was found to reverse the vincristine resistance of the above cells. Moreover, only PTX, but not other xanthine, increased the accumulation of [3H]vincristine by L1210/VCR cells. Thus it may be concluded that PTX-induced reversal of vincristine resistance could not be explained from the point of known pharmacological effects of PTX that are common for other xanthines such as inhibition of phosphodiesterase activity, calcium mobilizing effect, inhibition of tumor necrosis factor alpha (TNF), etc.

Serum tumor markers in chronic liver disease.

Tumor markers have been used for the evaluation of various malignancies though the existence of false positive results in some benign diseases is known. In this study, several established markers including carcinoembryonic antigen, alpha fetoprotein, beta human chorionic gonadotropin, ferritin, CA 19-9 and CA 125 were measured in 60 patients with chronic active hepatitis, 70 patients with cirrhosis and 40 normal subjects in order to evaluate the rate of false elevation of tumor markers in chronic liver disease. Prostate specific antigen and prostatic acid phosphatase levels were also measured in male patients and controls. Serum alpha fetoprotein levels were found elevated in 20% of patients with cirrhosis. The serum CA 19-9 level showed significant elevation in chronic active hepatitis (32%) and cirrhosis (44%). Increase in CA 125 concentration was also remarkable in chronic active hepatitis (23%) and especially in cirrhosis (74%). These results indicate that it is necessary to consider the presence of high false positivity rate of CA 19-9 and CA 125 during clinical interpretation of tumor markers in patients with chronic liver disease.

DNA ploidy in neuroblastoma.

Neuroblastoma is perhaps the most heterogeneous childhood cancer in terms of clinical behavior. Stage of disease, age at diagnosis, levels of urinary catecholamine excretion, N-myc amplification, and DNA ploidy have been found to be significant prognostic factors. The aims of this combined retrospective-prospective study are to verify the prognostic significance of DNA ploidy and to show its correlation with other prognostic signs. Thirty six fresh and thirty three paraffin embedded samples from patients with histologically confirmed neuroblastoma (41 prior to receiving any chemotherapy) were available for flow cytometry DNA analysis. Our results showed that the maturation induced during chemotherapy could give rise to aneuploidy therefore we analyzed the associations between the DNA ploidy and other prognostic markers only in patients examined before chemotherapy. There were no significant correlations between DNA ploidy and urinary catecholamine metabolites levels or tumor localization. DNA aneuploidy was significantly more frequent in patients with lower clinical stage, lower age at diagnosis, and without N-myc gene amplification. Patients with DNA aneuploid neuroblastomas died less frequently than patients with DNA diploid tumors. There were no significant associations among the S-phase or proliferation fraction and other prognostic factors.

Study of a new tumor marker, CYFRA 21-1, in squamous cell carcinoma of the cervix, and comparison with squamous cell carcinoma antigen.

The diagnostic value of a new tumor marker, CYFRA 21-1, was studied in the blood samples collected from 22 controls, and 87 pre-treatment patients with squamous cell carcinoma of the cervix. Sensitivity and specificity of CYFRA 21-1 was compared with those of squamous cell carcinoma antigen (SCC) measured in the sera of the same patients. Serum CYFRA 21-1 levels were higher in patients with squamous cell carcinoma than in controls (p < 0.05), and correlated with FIGO stage (Stage IIb-IV vs. Stage Ib-IIa, p = 0.0477). Using 2.5 ng/ml as cut-off value, elevated CYFRA 21-1 levels were found in 13.6% of controls, 34.8% of patients with Stage Ib-IIa squamous cell carcinoma of the cervix, and 63.5% of patients with Stage IIb-IV squamous cell carcinoma of the cervix. However, there was less sensitivity and specificity of CYFRA 21-1 than those of SCC in detecting squamous cell carcinoma of the cervix. CYFRA 21-1 may not be a better tumor marker than SCC for squamous cell carcinoma of the cervix.

The role of iron supply in the regulation of 5-aminolevulinate synthase mRNA levels in murine erythroleukemia cells.

Mouse erythroleukemia (MEL) cells transformed by Friend virus and induced to undergo erythroid differentiation by treatment with hexamethylenebisacetamide (HMBA) increase erythroid specific 5-aminolevulinate synthase (ALAS-E) mRNA levels by 4-15-fold and decrease "housekeeping" 5-aminolevulinate synthase (ALAS-N) mRNA levels by 1.2-1.4-fold. Iron affects translation of (ALAS-E) mRNA but nothing is known about its effect at the pretranslational level of the expression of (ALAS-E) mRNA. The aim of this study was to examine the effect of iron on the synthesis of (ALAS-E) mRNA and (ALAS-N) mRNA. This effect was compared with the effect of iron on the iron on the synthesis of H-ferritin and transferrin receptor mRNAs. Incubation of uninduced or induced MEL cells with iron chelator pyridoxal isonicotinoyl hydrazone (PIH) or desferrioxamine (Desferal) and 3H-uridine decreased the level of the 3H-labeled (ALAS-E) mRNA. The treatment with either diferric transferrin or Fe-PIH increased the level of the 3H-labeled (ALAS-E) mRNA. The opposite effect was observed on the level of the 3H-labeled (ALAS-N) mRNA. These findings indicate that iron might play its role also at the pretranslational level of the expression of ALAS-E or in the stability of (ALAS-E) mRNA.

Sequential combination of radio-chemotherapy (5-fluorouracil, cis-platinum and irradiation) in the management of locally advanced head and neck cancers.

Thirty-seven previously untreated patients with advanced, inoperable head and neck were treated with a sequential courses combining hypofractionated irradiation with chemotherapy (5-fluorouracil and cis-platinum). Each course was repeated every 4 weeks. Tumor response was evaluated and for 15 patients (41%) with a partial or complete regression after 3 radio-chemotherapy courses conventional radiotherapy was added. Eleven percent of all patients were in complete remission at the end of a treatment. This tumor response rate and the 50% rate of pain subside after first course for symptomatic patients contributed for a good palliative effect in the present study. However, the median survival of 7.2 months was considered unsatisfactory.

Tamoxifen as primary treatment of breast cancer in elderly patients.

The authors report on a series of 120 elderly (age over 69) women with primary breast treated with tamoxifen alone. Treatment schedule was 160 mg on day 1 followed by a daily maintenance dose of 20 mg. Compliance to treatment was optimal and side effects were minimal. The best results achieved after at least six months of treatments were complete response in 12, a partial response in 46 and minor response in 10 patients, whereas stable disease or progression was observed in 43 or 9 patients, respectively. Response duration was limited and progression was increasingly observed over time. After 6, 12, 24, 36, 48 and 60 months the proportion of subjects still showing response to treatment was 43%, 57%, 56%, 46%, 32% and 31%, whereas progression rate was 7%, 12%, 25%, 39%, 55% and 60%, respectively, the difference being accounted for by patients with stable disease. As determined in a subset of 27 subjects, treatment response was strongly associated with immunocytochemically assessed tumor estrogen receptor content, progression being 100%, 43% or 6% in subjects with 0%, 30-60% or > 60% immunostained cells, respectively. These results do not support primary hormone therapy as a current alternative to surgery, which should be the standard treatment in otherwise healthy elderly patients with operable breast cancer. When surgery is specifically contraindicated, hormone treatment should be proposed as an alternative only in subjects with high tumor estrogen receptor content.

Management of germ cell testicular cancer with pulmonary metastases.

Twenty eight patients with germ cell testicular cancer pulmonary metastases received primary chemotherapy including bleomycin, etoposide, and cisplatin (BEP). Complete response was achieved in 21 (75%) patients, in 11 of them CR was achieved following chemotherapy alone. Postchemotherapy surgery of residual mass was performed in 12 (42.9%) patients with normalized serum tumor markers. Retroperitoneal lymph node dissection was performed in one patient, pulmonary surgery in four, and both postchemotherapy treatments in 7 patients. Overall cure rate was 89.3%, 26 (92.9%) patients are still alive at a mean follow-up of 19.7+ months (range, 3-34+ months) after the treatment start. Two (7.1%) patients died: one of them due to disease progression during chemotherapy, and the second one due to postoperative complication (acute respiratory failure). Relapse of disease was observed in one patient 21 months following CR achievement, and sequential chemotherapy was introduced. Authors recommend surgical remove of all radiologically detected residual deposits, because the available imaging methods are not adequate for determining the histologic composition of residual mass, which is decisive for further therapy and has prognostic value.

A comparative study of preoperative B-V-M-M chemotherapy and irradiation in advanced squamous cell cancer of the oral cavity.

From January 1976, 50 patients with squamous cell cancer of the head and neck were treated with telecobalt preoperative irradiation followed by appropriate surgery. Another group of 50 patients, who matched in risk factors and stage of disease, were treated with preoperative chemotherapy and surgery. Chemotherapy consisted of bleomycin, vincristine, mitolactol and methotrexate. All patients received 3 courses. Surgery was performed 2-3 weeks post-chemotherapy or 4-6 weeks postradiotherapy. Forty-four percent of the patients in the radiotherapy group showed recurrences, while 30% of the patients had recurrence in the chemotherapy group. The overall 3-year survival rate was 66% in the chemotherapy group and 57% in the radiation therapy group, with no statistical difference.

Origin of choriocarcinoma in previous molar pregnancy proved by DNA analysis.

A 17-year old woman had in a short time period (seven months) a very exciting reproduction history. Molar pregnancy in December 1993, choriocarcinoma in January 1994 and induced abortion in June 1994. DNA analysis proved the origin of the choriocarcinoma in the previous molar pregnancy.

Dietary factors and risk of prostate cancer in Poland. Results of case-control study.

The relationship between cigarette smoking, vodka drinking and consumption of 44 food items typical of the Polish diet were analyzed in a case-control study performed in Cracow, Poland, among 76 cases of prostate cancer and 152 controls. Cigarette smoking and drinking of vodka were not significantly influencing the prostate cancer. The men who ate smoked or fried fish or liver at least once per week had almost half of the risk of prostate cancer of the men who ate those food rarely.

Soluble intercellular adhesion molecule-1 in colorectal cancer and its relationship to acute phase proteins.

Increased concentrations of soluble intercellular adhesion molecule-I (ICAM-1) have been reported in a number of diseases including cancer. This study was undertaken to evaluate soluble ICAM-1 in colorectal cancer and its relationship to an unspecific acute phase response. Fifty six patients (25 with advanced colorectal cancer and 31 out-patients after radical surgical treatment) were included. Soluble ICAM-1 was measured by enzyme immunoassay. Four acute phase proteins (C-reactive protein, acid alpha 1-glycoprotein, haptoglobin and ceruloplasmin) were estimated by immuno-nephelometry. No significant increase of soluble ICAM-1 could be demonstrated in the patients compared to a control group (median 273 ng/ml vs. 270 ng/ml). Furthermore, patients with advanced colorectal cancer did not demonstrate elevated soluble ICAM-1 compared to follow-up out-patients. Patients with present acute phase response as determined by C-reactive protein were shown to have increased soluble ICAM-1 compared to patients without acute phase reaction. Using other acute phase proteins no difference for soluble ICAM-1 has been shown. Our data suggest an association between acute phase response and increased ICAM-1 in patients with colorectal cancer which should be considered when the diagnostic and/or prognostic usefulness of soluble ICAM-1 is to be evaluated.

Antioxidant enzymes and lipid peroxidation in patients with multiple myeloma.

Reactive oxygen species and other free radicals are known to be the mediators of phenotypic and genotypic changes that lead from mutation to neoplasia. The imbalance in tumor cell antioxidant defense mechanism can influence also the sensitivity to cytoreductive therapy. In erythrocytes it can result to hemolysis which is one of the pathogenetic mechanisms of anemia in cancer patients. Parameters of lipid peroxidation (malondialdehyde-MDA) and antioxidant enzymes here represented by superoxide dismutase (SOD) and glutathione peroxidase (GPx) in multiple myeloma (MM) have been investigated. Nine patients of various clinical stages and activities of the disease were studied. Significantly higher concentrations of total MDA in plasma (1.20 +/- 0.24 mumol/l vs. 0.64 +/- 0.22 mumol/l, p < 0.0001) as well as in erythrocytes (2.72 +/- 0.81 mumol/l vs. 1.03 +/- 0.44 mumol/l, p < 0.0001) were found comparing to the control group. The levels of free MDA in plasma (0.31 +/- 0.09 mumol/l vs. 0.49 +/- 0.17 mumol/l, p < 0.05) and in erythrocytes (0.29 +/- 0.20 mumol/l vs. 0.59 +/- 0.22 mumol/l, p < 0.001) were decreased in myeloma patients. Significantly lower activities of GPx (19.17 +/- 4.07 U/g Hb vs. 23.26 +/- 3.61 U/g Hb, p < 0.05) and SOD (1882.46 +/- 181.73 U/g Hb vs. 2347.13 +/- 502.51 U/g Hb, p < 0.05) in erythrocytes were found. We did not observe evident relationship between the concentration of MDA or the activities of SOD and GPx and either the stage of the disease, or the level and the type of paraprotein. These results propose possible role of free radicals with reduced antioxidant activities of SOD and GPx in multiple myeloma.

PCR amplification of DNA from archival specimens. A methodological approach.

The experiments were designed to assess whether DNA could be recovered from archival, fixed, paraffin embedded specimens, 1-39 years old, for use in PCR and Southern blot analyses. Specimen fixed in either 10% formalin, Carnoy's or AMeX fixative produced almost equally abundant 318 bp long beta-actin fragment, after 2 x 120 min deparaffinization time, proteinase K DNA extraction and 40, or more, amplification cycles. Prolonged deparaffinization time and phenol/chloroform extraction did not influence DNA quality for PCR. Formalin fixed tissues can be successfully used for DNA/PCR of shorter fragments (318 bp) even if they are up to 39 years old. Longer fragment (720 bp) was successfully amplified from 1 and 10 years old specimens, also investigated whether DNA suitable for hybridization studies could be prepared from fixed tissue. Formalin caused irreversible DNA damages which were greater with prolonged fixation time making it unsuitable for hybridization studies, but still suitable for PCR amplification. Carnoy's and AMeX fixation resulted in consistently high yield of high molecular size DNA suitable for use in hybridization studies and PCR respectively. DNA from Papanicolaou stained smears was successfully amplified by PCR as well. Of all stains used in the preparation of smears, only eosin was detectable as a greenish band in agarose gels under ultraviolet illumination.

Presence in bovine fetal serum of the protein antigenically related to p65-tumor associated antigen: its isolation and polyclonal antibody production.

Monoclonal antibodies raised to the 65-kDa tumor-associated protein (p65) isolated from a human breast cancer cell line have been used to detect an antigenically related protein (p65-like) present in fetal bovine serum (FBS) by Western blot analysis. We have isolated the p65-like protein from FBS by isoelectrofocusing (IEF) on native gels followed by electrophoresis in 12.5% polyacrylamide gel containing 0.1% SDS (SDS-PAGE). Immunostaining with anti-p65 monoclonal antibody of fetal bovine serum fractions separated by electrophoresis on cellulose acetate membrane revealed that the p65-like protein had a location similar to one of gamma-globulin. This protein migrates as a single band upon electrophoresis in SDS-PAGE and had four isoforms which migrate as two doublets with pI's of approximately 5.0 and 5.3.

Changes of Ki67 index of various tumors during radiation therapy.

This study investigates the changes in the Ki67 labeling index during radiotherapy (RT) of seven patients with primary breast carcinoma, one patient with metastatic bronchial squamous cell carcinoma and one patient with a paraumbilical deposit of a large bowel adenocarcinoma. The material was taken by fine needle aspiration either from the primary tumors or the metastases. In four patients with primary breast carcinoma, we observed a drop in the Ki67 labeling index after 24 h (2 Gy), but an increase after 18 days (26 Gy). In one patient the pattern was different. An increase after the initial fraction was followed by a decrease after 18 days of radiotherapy. In two patients with primary breast carcinoma, where the second sample was aspirated after 72 h (3 fractions of RT, 6 Gy), the values of Ki67 labeling indexes before RT were higher than after RT. In two patients where the material was aspirated from metastatic deposits, we observed an increase of Ki67 labeling index after 24 h (2 Gy). Possible explanations, including repopulation following the suppression of the synthesis in the originally active clonogens after radiation injury, are discussed.

DTIC vs. IFN-alpha plus DTIC in the treatment of patients with metastatic malignant melanoma.

In our study we evaluated and compared the therapeutic success in 70 patients with cutaneous metastatic malignant melanoma (MM) treated at the Institute of Oncology in Ljubljana during the period 1985-1994. Twenty-nine patients received DTIC in a single 800 mg/m2 i.v. dose (Group 1) and 41 patients were receiving i.m. applications of IFN-alpha in 2 MU daily doses from days 1 to 4, completing the treatment with a DTIC application on day 5, given at the same dosage as in Group 1 (Group 2). The applications were repeated in three-week intervals until progression, or-in the case of a complete response-for up to 6 months. The rate and median duration of treatment response were higher in the group of patients treated by IFN-alpha plus DTIC (17% vs. 27%; 2.7 vs. 6.1 months), though the difference was not statistically significant. The survival of responders was either significantly higher (Group 2: p = 0.0007) or borderline-significantly higher (Group 1; p = 0.078) than that of non-responders. These patients also had significantly longer median survival (Group 1: 13.7 vs. 5.1 months, p = 0.019; Group 2: 19.3 vs. 4.9 months, p = 0.0003). The patients treated with IFN-alpha plus DTIC survived significantly better than those treated with DTIC alone (p = 0.043). There were no differences in the median duration of survival between both groups (6.6 vs. 6.7 months), and neither in the median duration of survival of responders (13.7 vs. 19.3 months) or non-responders (5.1 vs. 4.9 months) from both groups. The toxicity of combined therapy was higher than that of chemotherapy alone, though it was still moderate and acceptable. In view of our results, the addition of IFN-alpha to DTIC has shown an advantage over DTIC along.

Glutathione peroxidase, reduced glutathione, superoxide dismutase and catalase in red cells of patients with hairy cell leukemia.

Red cell antioxidant enzymes have been recently studied in malignant lymphomas and the results are controversial. Hairy cell leukemia is a rare chronic lymphoproliferative disorder originating probably in a pluripotent stem cell. In the present study, glutathione peroxidase (Gpx), reduced glutathione (GSH), superoxide dismutase (SOD), and catalase were measured in a homogeneous group of patients with untreated hairy cell leukemia and normal controls. Glutathione peroxidase, catalase and SOD activities were significantly lower in patients than in normals. GSH was not significantly different in patients compared to controls. There was no correlation between Gpx, GSH, SOD, and catalase and hemoglobin; reticulocytes, leukocytes, hairy cells, platelets number or splenomegaly. Taken together these data suggest a decreased activity of red cell antioxidant enzymes in hairy cell leukemia and support a pluripotent stem cell defect of these abnormalities.

Serum prostate-specific antigen in healthy Chinese men: establishment of age-specific reference ranges.

The purpose of this study is to establish the normal range of serum prostate-specific antigen (PSA) and to evaluate the influence of age in a population of healthy Chinese men. Subjects with PSA concentration of less than 4.0 ng/ml and normal digital rectal examination (DRE) or PSA level greater than 4.0 ng/ml and/or abnormal results of DRE with negative biopsy were defined as clinically free of prostate cancer. A total of 1008 men aged between 21 and 80 years of age fulfilled the criteria of establishing the upper limits for PSA levels. The population was grouped by age decades. The 95th percentile was determined as the upper limit of normal (reference range) for each 10-year age group for the serum PSA concentration. Our data showed that the upper limit of normal (95th percentile) for the serum PSA concentration increases with age. It is 1.92 ng/ml for men 21 to 30 years of age. 1.85 ng/ml for men 31 to 40 years of age, 2.59 ng/ml for men 41 to 50 years of age, 3.31 ng/ml for men 51 to 60 years of age, 5.03 ng/ml for men 61 to 70 years of age and 5.73 ng/ml for men 71 to 80 years of age. In conclusion, the serum PSA concentration is directly correlated with patient age. The age-specific reference ranges should be established so as to increase diagnostic specificity in older men and increase diagnostic sensitivity in younger men and then the PSA assay can become more efficient as a screening test for prostatic cancer.

SPE-HPLC determination of catecholamines using an affinity principle.

Solid-phase extraction (SPE) with the affinity chromatography principle was applied for high performance liquid chromatography (HPLC) determination of catecholamines in patients urine samples. Electrochemical amperometric detection (ED) was used for all HPLC analyses and both analytical and microbore columns were tested for optimal chromatographic resolution of all analyzed catecholamines. Capacity ratio k' and detection limits were evaluated for all columns used. Precolumn affinity chromatographic procedure of catecholamines with diphenylboronic acid (DPBA) in alkaline medium improved their retention on different commercial SPE reversed-phase cartridges. After clean-up and preconcentration step the complex catecholamine-diphenylboronic acid was degraded by acidic pH and eluted from SPE cartridge. After SPE affinity step catecholamines were analyzed by HPLC-ED. The optimal elution solutions was chosen also as suitable SPE cartridges. Extraction recoveries of catecholamines were 89.6-93.3% with relative standard deviation RSD = 3.8-4.3%. Total analysis time was 30 min including 15 min for the preseparation procedure.

Polarographic reduction of some triphenylmethane dyes and their potential carcinogenic activity.

Polarographic reduction of some triphenylmethane dyes in strictly anhydrous solutions was studied in the absence and presence of alpha-lipoic acid. The values of the half-wave potentials E1/2 and the parameter of potential carcinogenicity tg alpha were determined for all compounds studied. The current value of the first diffuse polarographic wave was increased in the presence of alpha-lipoic acid for all of the compounds. The increase was linear and depended on the alpha-lipoic acid concentration in anhydrous solutions. The highest determined value of tg alpha belonged to crystal violet (0.420) and to methyl violet 2B (0.440). The values indicate potential carcinogenic activities of the respective triphenylmethane dyes.