The use of epidemiology for screening and early detection of cancer: examples from Austria.

Reduction of mortality rates and the costs of screening programs have sometimes been discussed on basis of certain therapeutic nihilism especially concerning the treatment of some cancer localizations. Interest of an epidemiological institute should, however, concern specific regions with high cancer risk and in these areas, well defined by epidemiological investigations, screening projects should be performed. A screening program initiated in Oberpullendorf, a district in the federal country of Burgenland in Austria resulted in a rate of 53% Dukes A colorectal cancer stages (0% Dukes D) compared to 27% Dukes A stage (19% Dukes D) without screening. As regards the lung cancer screening programs, these are controversial, due to high costs and no evident influence on mortality rates. The mortality rate for Vienna is about 20% above the Austrian average. In women the discrepancy between the capital and the federal countries is especially high, as in Vienna 44% more women die on lung cancer as compared to the Austrian average. Some promising results in the treatment of lung cancer, especially in small cell carcinoma and combined treatment by chemo- and radiotherapy might justify a screening program for lung cancer. The highest increase of mortality in lung cancer is found in patients at the age of 75 and older. Those patients, however, suffer from other diseases too, therefore "screening programs for polymorbidity" should be discussed.

The role of platinum uptake and glutathione levels in L1210 cells sensitive and resistant to cisplatin, tetraplatin or carboplatin.

Three L1210 murine leukemia variants resistant to cisplatin, tetraplatin or carboplatin were compared with their sensitive parent line for differences in platinum accumulation, efflux and glutathione (GSH) content. All three resistant lines had reduced platinum accumulation compared with the sensitive line following exposure to each of the three drugs. There was no difference in the efflux of platinum between the sensitive and resistant lines indicating that reduced platinum accumulation was due to impaired uptake mechanisms. However, in cell survival experiments it was apparent that the resistant lines could tolerate higher intracellular platinum levels than the sensitive line. Intracellular GSH measurements revealed comparable levels in all but the L1210/carboplatin line which had a 2-fold elevation during the first 24 hours of culture. However, depletion of intracellular GSH with buthionin sulphoximine over a 72 hour period did not sensitize the resistant lines to the drugs. These studies have demonstrated that impaired platinum uptake is one of the mechanisms contributing to platinum drug resistance, while perturbations in GSH levels do not appear to play a role in these cell lines.

Collateral resistance or sensitivity of human larynx carcinoma HEp2 cells resistant to cis-dichlorodiammineplatinum (II) or vincristine sulfate.

Human larynx carcinoma HEp2 cells in tissue culture were repeatedly treated with two chemotherapeutic agents which exhibit distinct mechanisms of action and which differ in mechanisms underlying resistance: cis-dichlorodiammineplatinum (II) (cis-DDP) and vincristine sulfate (VCR). Two schedules of resistance development were used: Acute (cells repeatedly treated with drug for 1 hour in serum-free medium: CA15 and VA15 cells), and continuous (repeatedly treated for 24 hours in complete medium: CK15 and VK15 cells). The sensitivity of cis-DDP resistant (CA15 and CK15 cells) and VCR resistant (VA15 and VK15 cells) sublines to cis-DDP VCR, methotrexate (MTX) and gamma rays was determined by survival assay. The results showed that all sublines became resistant to MTX. Cis-DDP resistant cells became cross-resistant to VCR too. Of VCR resistant sublines, only VK15 cells changed (increased) their sensitivity to cis-DDP. Resistant sublines (except CA15) became sensitive to gamma rays. The degree of resistance or collateral resistance/sensitivity depended on the chemotherapeutic agent used for selection and on the schedule of resistance development. The present data support the idea of complexity of resistance phenomenon. They also emphasize the difficulty and importance of judicious choice of agents that should be given in combined therapy of tumors.

Chemotherapy of testicular cancer: 10-year experience.

A total of 250 patients with germ cell testicular tumors were treated by PVB chemotherapy between 1982 and 1992. Mean age of patients was 28.9 years (range 15-52). Thirty-four patients in clinical Stage II (11 patients IIA, 13 patients IIB, and 10 patients IIC) underwent primary retroperitoneal lymphadenectomy (RPL) with subsequent chemotherapy. They were followed-up for a mean of 106.3 months (range 85-125). CR was achieved in 30 patients (88.2%). Three patients relapsed. Twenty-seven patients (79.4%) are alive with no evidence of disease (NED) after a minimum of 5 years since the start of therapy. One hundred and twenty-two patients underwent primary chemotherapy for clinical Stages IM (15 patients), IIA (31 patients, IIB (48 patients) and IIC (28 patients) with RPL in cases with residual mass in the retroperitoneum. They were followed-up for a mean of 47.7 months (range 6-122). CR was achieved in 115 patients (92.7%) (75 of them received chemotherapy alone, 40 patients achieved CR following combined cytostatic-surgical treatment). Eleven patients relapsed. One hundred and nine patients (89.3%) are alive with NED. Ninety-four patients in Stages III and IV (8 patients III, 86 patients IV) underwent primary chemotherapy with additional surgical removal of residual metastases. They were followed-up for a mean of 50.5 months (range 6-125). CR was achieved in 65 patients (69.1%) (32 of them received chemotherapy alone, 33 patients achieved CR following combined cytostatic-surgical treatment). Eleven patients relapsed. Fifty-seven patients (60.6%) are alive NED. There were 11 patients with advanced germ cell testicular cancer (Stages IIC and IV) who underwent initial PVB chemotherapy without previous orchiectomy. Delayed orchiectomy was done simultaneously with surgical removal of residual mass in the retroperitoneum or in the lungs or at completion of chemotherapy alone. The toxicity of chemotherapy was moderate. There were drug-related deaths in ten patients (4%).

Polarographic reduction and carcinogenic index tg alpha of 5-aza nucleosides possessing antileukemic activity.

Polarographic behavior of arabinosyl-5-azacytosine (ara-AC) and 5-azacytidine in anhydrous dimethylformamide and in Britton Robinson buffer is described. 5-Azacytidine and ara-AC underwent two-electron reduction under the polarographic conditions used. Carcinogenic index tg alpha estimated in the presence of alpha-lipoic acid was 0.295 for 5-azacytidine and 0.275 for ara-AC.

Relation of some cytochemical activities to the expression of CD34 marker in acute leukemia.

The blast cells of peripheral blood and bone marrow of eleven acute leukemia patients with CD34 surface membrane antigen expression were investigated by immunophenotyping, cytochemistry and biochemical analysis. CD34 expression is a characteristic marker of very immature leukemic cells. Nine of eleven CD34+ acute leukemia cases had the immunophenotype of poorly differentiated myeloid cells. CD34+ blasts of two patients expressed the phenotype of very early lymphoid differentiation. Significant correlation was found between the expression of HLA-DR and CD34. The absence of CD10 antigen expression was observed in all cases of CD34+ myeloid blasts. The strong activity of SBB along with simultaneous absence of light microscopy MPO staining, 5'NT and moderate BG reactivity were characteristic enzyme features of CD34+ myeloid cells. The enzyme features of more mature stages of myeloid differentiation lacked. In lymphocytes, the CD34 expression correlated with CD10 antigen positivity and 5'NT activity. The activity of ADA was higher than that of PNP in all cases of CD34+ acute leukemia blast cells. The SBB, 5'NT and BG reactivity in correlation to CD34 antigen expression in acute leukemia may be an additional marker of very early stages of differentiation, either lymphoid or myeloid. The contribution to clinically important differential diagnosis of immature acute leukemia is discussed.

Modulation of protectin (CD59 antigen) cell surface expression on human neoplastic cell lines.

The ability of cytokines (IFN alpha, IFN gamma, TNF alpha, IL-1 alpha, IL-6), all-trans retinoic acid, 1,25(OH)2-vitamin D3 and the tumor promoting phorbol ester TPA to regulate cell surface expression of protectin (CD59 antigen) on human hematopoietic and non-hematopoietic neoplastic cell lines was examined with the aid of immunocytofluorometric measurements. The tumor promoting phorbol ester TPA induced a marked up-regulation of protectin in all examined cell lines with the exception of promyelocytic leukemia HL-60, where TPA significantly decreased protectin cell surface expression. All-trans retinoic acid weakly down-regulated cell surface protectin on K-562, while 1,25(OH)2-vitamin D3 produced such effect on HL-60 cells. None of the examined cytokines induced a significant protectin down-regulation in the examined cell lines.

Contribution of radiotherapy to the management of malignant melanoma. A ten year experience at the University of Illinois Hospital in Chicago.

Fraction size in radiotherapy of malignant melanoma remains a point of controversy. Among 139 patients treated at the University of Illinois Hospital in 1979-1988, 36 were considered potentially curable (not counting ocular melanomas); 20 were treated by the Princess Margaret Hospital (PMH) hypofractionated schedule using 800 cGy per fraction and achieved a permanency of local control lasting > 6 months since the beginning of radiotherapy in 10/22 (45.5%) courses. Comparable results were obtained in 11 patients treated by standard fractionation to at least threshold curative levels. A modification of PMH regimen in 5 patients (but with 13 courses) by decreasing fraction size to 400 cGy while keeping total dose and course duration unchanged, resulted in a 100% loss of focal control within 6 months. Patients considered incurable and irradiated by PMH schedule responded in 83% of courses compared to 51.4% response rate in patients irradiated with other schedules (except modified PMH regimen). Other aspects of melanoma management are analyzed.

Overcoming of vincristine resistance in L1210/VCR cells by several corticosteroids. Collateral sensitivity of resistant cells.

Five corticosteroids were tested to determine whether they are able to overcome the multidrug resistance of vincristine resistant mouse leukemia cells L1210/VCR. The most effective in reversing multidrug resistance were cortisone and dexamethasone, less effective as reversing agents were 11-deoxycorticosterone, 1-dehydrocortisone and hydrocortisone. By testing of collateral sensitivity of vincristine resistant cell line to these corticosteroids it was found that only 11-deoxycorticosterone and dexamethasone were toxic to multidrug resistant cells at doses much lower than required for toxicity to the drug-sensitive L1210/S cells. Using thin layer chromatography the polarity of tested corticosteroids was estimated, and good correlation was found between polarity of corticosteroids and their increased toxicity to vincristine resistant cells.

Amplification of HER-2(erbB-2/neu) oncogene as the most significant prognostic factor in a group of Russian breast cancer patients.

Amplification of HER-2(erbB-2/neu) oncogene was detected in 36 of 142 (25%) breast carcinomas (BC) RNA expression was examined in 42 carcinomas, in 10 of them overexpression was revealed. Amplification was matched by overexpression. No association was found between the increased number of HER-2(erbB-2/neu) copies and tumor size, lymph node involvement, stage of disease, age of onset, and estrogen and progesterone receptor level. HER-2(erbB-2/neu) amplification was shown to be of independent prognostic significance in the group of 32 BC patients with sufficient follow-up (more than 40 months). Six of 7 HER-2(erbB-2/neu) amplification-positive patients and only 2 of 25 HER-2(erbB-2/neu) amplification-negative ones relapsed (p < 0.00005).

Serum alpha-1-antitrypsin and alpha-1-antichymotrypsin after surgical treatment and during postoperative clinical course of human gastric cancer.

The usefulness of alpha-1-antitrypsin (A1AT) and alpha-1-antichymotrypsin (A1AChy) in monitoring the adequacy of surgical treatment in patients with gastric cancer was studied. In 32 patients A1AT and A1AChy serum levels were studied before and after radical or palliative surgery at regular intervals during a 3-50-month follow-up. After palliative surgery, A1AT and A1AChy concentrations were significantly higher than in patients treated radically who showed a decreasing tendency of both antiproteases. In patients with progressive disease after surgery an increase of both antiproteases was observed. A1AT and A1AChy levels in the remission group tended to decrease but the values did not normalize as compared to healthy subjects. It was suggested that the increasing levels of A1AT and A1AChy during chemotherapy after radical surgery for gastric cancer may point to the progress of the disease.

Chromosome sensitivity to bleomycin in patients with dominantly inherited and sporadic tumors.

G2 chromosomal sensitivity to bleomycin (30 micrograms/ml) was tested in PHA-stimulated lymphocytes of healthy subjects and in patients with familial and sporadic tumors. These were multiple endocrine neoplasias (MEN) types 1, 2A and 2B, familial medullar thyroid cancer, Recklinghausen neurofibromatosis type I, sporadic and hereditary malignant tumors, and a preleukemic disorder, the myelodysplastic syndrome. Control subjects were either young (15-20), middle-aged (28-49) or old (70-83 years). Cells from old healthy subjects and from subjects with MEN 1 showed increased sensitivity to clastogenic effects of bleomycin. All the remaining investigated groups were insignificantly different from controls. Our data suggest that in contrast with recessively inherited syndromes with chromosome instability the mutagen hypersensitivity, as evaluated by the extent of chromosomal damage, is not a feature of most dominantly inherited tumor syndromes.

Phorbol ester-induced modulation of cell surface antigens on U-937 cells in protein-free medium: effect of protein kinase and calmodulin inhibitors.

Phorbol ester (PMA) induced decrease in cell surface expression of CD4 antigen was potentiated on human monoblastoid cell line U-937 cultured and induced in protein-free culture medium. Down-regulation of the CD4 antigen was partially inhibited by the isoquinoline protein kinase inhibitor H7 and unaffected by the calmodulin inhibitor R24571 (calmidazolium). PMA-induced increase of the monocyte-associated CD14 antigen was dependent the presence of fetal bovine serum (FBS) in culture medium and partially abolished by calmodulin inhibitor R24571. H7 inhibitor partially abrogated this up-regulation in 4 out of 8 induction experiments. PMA-induced down-regulation of MHC class I antigen was found in both FBS-supplemented and protein-free culture media. This effect was partially inhibited by both H7 and R24571 inhibitors.

Neo-adjuvant chemotherapy with delayed orchiectomy in patients with advanced germ cell testicular cancer.

A total of 13 patients with advanced germ cell testicular cancer underwent initial PVB chemotherapy without previous orchiectomy. Complete response (CR) of metastases was observed in 5 patients following chemotherapy alone. The residual mass persisted in 8 patients (in 5 of them in the retroperitoneum, in two patients in the lungs only and in one patient in both localizations). The residual masses were removed surgically. There were no viable malignant tumors in the removed tissue on histological examination. Delayed orchiectomy was performed simultaneously with surgical removal of the residual mass in the retroperitoneum or in the lungs in 8 patients, and in 5 patients as a separate procedure in complete responders following chemotherapy alone. Residual viable tumor in the testis was found in three patients, necrotic or fibrotic tissue in 5 patients, and mature teratoma in 5 patients. In patients with advanced germ cell testicular cancer preference must be given to early beginning of intensive chemotherapy without tissue diagnosis of primary tumor by orchiectomy. Benefit of this therapeutic approach is the timely management of acute abdominal and/or pulmonary symptoms of life-threatening distant metastases.

c-Ha-ras BamHI RFLP in human urothelial tumors and point mutations in hot codons.

High-molecular-weight DNAs from 30 bladder and renal cell carcinomas (RCC) were isolated and the c-Ha-ras gene BamHI RFLP was examined. Amplification of c-Ha-ras with normal localization with regard to the size of alleles was found only in one case. One of the normally localized c-Ha-ras allele termed RCC c-Ha-ras of a length of about 6.6 kbp was cloned and an oncogene-activating point mutation was identified using two restriction enzymes. After comparison of CfrI and Cfr10I cleavage maps of RCC c-Ha-ras to complete nucleotide sequences of EJ/T24 c-Ha-ras oncogene and its normal counterpart, a point mutation was identified within codon 11 or 12. The use of CfrI and Cfr10I is of value for clinical practice in identification of point mutations in c-Ha-ras PCR product in neoplasia accompanied by somatic mutation of c-Ha-ras. The correlation among c-Ha-ras allele, amplification/loss, presence of point mutation and progression of neoplasia is discussed.

Plasma membrane enzymes in human breast carcinoma: relationship with serum hormones.

Alkaline phosphatase, 5'nucleotidase and gammaglutamyl transferase were studied in premenopausal and post-menopausal women with fibroadenoma and breast carcinoma tissues. The relationship of circulating estradiol (E2) and prolactin (Prl) with these enzymes was also investigated. All the three enzyme activities were found to be elevated in the breast carcinoma tissues of both pre- and postmenopausal groups. None of the membrane bound enzymes studied was altered in fibroadenoma breast tissues. The activities of all the three enzymes in breast carcinoma tissues were stimulated by the elevated level of serum Prl and further stimulated by a simultaneous increase in E2.

Multifactorial molecular mechanisms are involved in resistance of preirradiated human cervix carcinoma cells to cis-dichlorodiammineplatinum (II) and vincristine.

We previously found that human cervix carcinoma HeLa cells irradiated with multiple fractions of gamma rays (0.5 Gy daily, five times per week over 6 weeks) become resistant to cis-dichlorodiammineplatinum(II) (cis-DDP), methotrexate (MTX) and vincristine (VCR), but retain the same sensitivity to gamma rays or UV light. In the present report attempts were made to elucidate the mechanisms by which these cells have acquired resistance to cis-DDP and VCR. The sensitivity to different drugs was measured by modified MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method. Neither buthionine sulfoximine (BSO) nor ethacrinic acid were able to reverse the resistance of preirradiated cells to cis-DDP. Therefore, neither the increased levels of glutathione nor glutathione transferase seem to be involved in resistance to cis-DDP. Preirradiated cells did show resistance to cadmium, indicating the increased levels of metallothioneins in these cells. Resistance of preirradiated cells to vincristine was abolished by the addition of verapamil, indicating that resistance to this drug may depend on the increased expression of plasma membrane P-glycoprotein. It was concluded that mechanisms of resistance of preirradiated cells to cytostatics are multifactorial and involve at least the increased levels of metallothioneins and changes in the plasma membrane. Acquired resistance to cytotoxic drugs induced by preirradiation may be the reason for the reduced response to these drugs after radiation treatment of certain tumors.

Effect of 1-alkylpyrrolidine N-oxides on energy metabolism of cancer cells.

The main purpose of the present investigation was to study the effect of a homologous series of 1-alkylpyrrolidine N-oxides on ATP-producing processes in Ehrlich ascites and L1210 murine leukemia cells. The effect on aerobic glucose consumption, lactic acid formation, content of total (T-SH) and non-protein thiol groups (NP-SH), endogenous respiration and the level of ATP in tumor cells incubated in vitro was investigated. 1-Tetradecylpyrrolidine N-oxide (TPNO), one of the most active compounds, immediately after addition to the suspension of Ehrlich cells in an ice bath, decreased the level of ATP to the same extent over the whole concentration range. After 2 h incubation at 37 degrees C the drop in the ATP level was lower. The decrease in ATP level might be explained through the interaction of the amine oxide with the cell membrane integrity.

New tumoricidal semisynthetic ether phospholipid, plasmanyl-(N-acyl)ethanolamine (PNAE(s)) and enhancement of its tumoricidal activity by calcium ions.

Recently we described a novel, nontoxic, natural ether phospholipid with selective antitumor activity, 1-0-alkyl-2-acyl-sn-glycero-3-phospho-(N-acyl)ethanolamine, i.e., plasmanyl-(N-acyl)ethanolamine (PNAE), isolated from ischemic tissue of chick embryo. The chemical structure of PNAE has been confirmed by partial synthesis. The semisynthetic preparation PNAE(s), 1-0-octadecyl-2-oleoyl-sn-glycero-3-phospho-(N-palmitoyl)ethanolamine, exhibited tumoricidal activity against human tumor cells T24 and mouse sarcoma cells Mcll as well as in vivo against murine sarcomas S-180 and Mcll. PNAE(s) selectively destroyed tumor cell membranes as has been demonstrated by scanning electron microscopy. The antitumor activity of PNAE(s) in vitro and in vivo was significantly enhanced by Ca2+ ions. These results may be explained by selective damage of tumor cell membranes by PNAE(s), increasing also the tumor cell membrane permeability for exogenous Ca2+ ions and altering the intracellular calcium homeostasis in tumor cells. By increased concentration of Ca2+ in tumor cell cytosol selective damage and lysis of tumor cells was induced. The combined use of parenteral administration of PNAE(s) and peroral doses of calcium gluconate may provide a new approach to enhancement of antitumor activity of PNAE(s) by Ca2+ in a very selective and nontoxic antitumor therapy.

Evaluation of 2-(methylaminosulfonyl)-1-(arylsulfonyl)-1-methylhydrazines as anticancer agents.

Seven new 2-(methylaminosulfonyl)-1-(arylsulfonyl)-1-methylhydrazines were prepared. The anticancer activity of these compounds was assessed in murine Ehrlich ascites carcinoma (EAC) by in vivo screening. Moderate in vivo activity in EAC was exhibited by three compounds. All of them were screened in vitro against a battery of human tumor cell lines at the National Cancer Institute (NCI), USA. One of them, compound 3a has displayed highly significant specificity in the renal tumor cell line RXF 393. These three compounds were also assessed for in vitro anti-HIV activity at the NCI, however, they have not reached the criteria of significant activity. The alkylating activity of the compounds was determined by measuring the absorbance of the alkylated product of 4-(4-nitrobenzyl)pyridine. It has been found that they are capable of acting as chemical alkylating agents.

Effect of three biological response modifiers on chemical carcinogenesis in mice.

The modulation of chemical carcinogenesis by three biological response modifiers was assessed in a mouse model. CBA mice given 20-methylcholanthrene s.c. were treated with peptidoglycan monomer, azure B and indomethacin for one month, either from day 0 or 75 after methylcholanthrene injection to assess their effects on tumor incidence (on days 150 and 300), time of tumor appearance, time of death, and duration and dynamics of tumor growth. All three agents significantly influenced some of the parameters of tumor growth, except tumor incidence on day 300. Highly significant sex differences in tumor appearance and growth were observed. Tumors with late appearance grew faster in comparison to tumors with early appearance. The data presented indicate that the effectiveness of anti-cancer body defense mechanisms can be best defined by the time of tumor appearance.

Vitamin E--its status and role in leukemia and lymphoma.

A comparative study has been performed on the relationship between vitamin E and immunofunction in normal and malignant condition in human and murine systems. Further, the effects of supplemental vitamin E on tumor take, host survival and tumor growth have been studied in a transplantable lymphoma in mice. Vitamin E was assayed in serum samples from normal subjects and from patients with leukemia and lymphoma by high performance liquid chromatography (HPLC). The murine group included Dalton's ascitic lymphoma (DL), Schwartz lymphoblastic leukemia (SVL) and Moloney lymphoblastic leukemia (MVL). Serum vitamin E was found to be lower than that of the normal controls in all cases of leukemia and lymphoma both in human and animal system. The levels of immunoglobulins (IgG and IgM) were found to be higher in mice with leukemia and lymphoma. Supplementary vitamin E administered at the initial phase of development of murine lymphomas reduced the rate of tumor growth, improved host survival and elevated serum vitamin E level. Vitamin E supplementation also activated specific mitogen induced blastogenesis of peripheral blood lymphocytes (PBL) and elevated serum IgG level. IgM remained unaltered and macrophage activity did not seem to be affected. The present findings indicated a low status of vitamin E in tumor bearing host and a beneficial effect of supplemental vitamin E on the host which was mediated by the host immune system.

HPLC analysis of optical isomers of leucovorin and methotrexate using achiral-chiral system.

Chiral high performance liquid chromatography was used for the enantiometric separation of leucovorin. The optimal separation conditions recommended after optimizing the mobile phase composition, flow rate, and temperature are described. Achiral reversed-phase chromatographic method was applied for the simultaneous separation of methotrexate and leucovorin in clinical samples. Column-switching system including achiral short pre-column and chiral analytical column based on immobilized bovine serum albumin were used for simultaneous determination of leucovorin and methotrexate patients treated at the National Cancer Institute.

Estimation of reduction of life-time risk of cervical cancer through one life-time screening.

The purpose of screening for cervical cancer is to prevent the appearance of invasive disease by the detection and treatment of precancerous lesions. Based on data from the developed countries, recommendations have been made regarding the age at which cytology screening should begin and the interval with which rescreening should be performed. However, these recommendations may not be applicable for a developing country like India due to human and financial resource constraints. WHO has recommended that for countries where resources are limited the aim should be to screen every woman at least once in her life-time at an appropriate age. Hence, it is essential to determine the age at which screening may be offered to derive maximum gains. With this in background, based on the data of national cancer registries, an attempt has been made for Indian population to estimate the reduction in the cumulative incidence rate of cervical carcinoma which can be achieved in a cohort of women aged 20-64 years by screening once in her life-time at an appropriate age. Our findings revealed that the first screening offered at 45 years of age could produce gains to the extent of 20-27%, while the screening offered at 50 or 55 years produced the maximum gains to the extent of 25-29%. However, keeping in view the produce years of lives lost in younger women, it may be reasonable to offer once in life-time screening to women aged 45 years.

Amelioration of doxorubicin cardiotoxicity by the bispiperazinedione ICRF-187 in women with advanced breast cancer: a preliminary report.

A novel cell growth-promoting factor identified in a B cell leukemia cell line, BALL-1.

A novel leukemia cell growth-promoting activity has been identified in the culture supernatant from a human B cell leukemia cell line, BALL-1. The supernatant from unstimulated cultures of the BALL-1 cells significantly promoted the growth of 16 out of 24 leukemia/lymphoma cell lines of different lineages (T, B and non-lymphoid) in a minimal concentration of fetal bovine serum (FBS), and of 5 out of 12 cases of fresh leukemia cells in FBS-free medium. The growth-promoting activity in the BALL-1 supernatant has been further characterized using FPLC chromatography, molecular weight (MW) sieve filtration and dialysis. The MW of the factor was less than 10 kDa. The growth-promoting activity was heat and acid stable and resistant to trypsin treatment. The factor isolated from the BALL-1 supernatant was distinct from known polypeptide growth factors with MW below 10 kDa, such as epidermal growth factor, transforming growth factor alpha, insulin-like growth factor I (IGF-I), IGF-II and insulin, as determined by specific antibodies and by cell growth-promoting tests. The factor in the BALL-1 supernatant did not promote the proliferation of normal human fresh peripheral blood lymphocytes or mouse fibroblast cell line, BALB/c3T3. In addition to the BALL-1 supernatant, a similar growth-promoting activity was found in the culture supernatants from 13 of 17 leukemia/lymphoma cell lines tested. The activity in these culture supernatants promoted the growth of leukemia/lymphoma cell lines in autocrine and/or paracrine fashions. These observations suggest that the low MW cell growth-promoting activity found in the BALL-1 culture supernatant is mediated by a novel factor which may well be responsible for the clonal expansion of particular leukemic clones.

Leukemia-associated marker combinations in acute leukemia suitable for detection of minimal residual disease.

In the absence of truly leukemia-specific antigen, antigen combinations were identified in leukemia cells that are absent or extremely rare among normal hemopoietic cells. Some of the studied combinations related to the simultaneous surface and cytoplasmic marker expression, others, expressed mainly on cell surface membrane, represented atypical or aberrant combinations. Comparing membrane (m) and cytoplasmic (c) antigen expression (followed in 23 acute leukemia cases), we observed that CD3 could be detected in cytoplasm in the majority of T-ALL cells, while was absent on cell surface membrane where simultaneous expression of more immature T cell markers, such as CD7 and CD5, could be detected. Combination of mCD7/cCD3 could be regarded as a suitable marker of individual T-ALL cells. In cases of B-precursors of acute leukemia cells, leukemia-related combination of mCD19/cCD22 was found, which could characterize a single leukemia cell. The cells in one of 11 AML followed cases were positive for CD13 in cytoplasm, but not on cell surface membrane, where CD33 and other myeloid antigens were expressed. The cells in another two AML cases were positive for CD11 in cytoplasm but not on cell surface membrane, where CD13 or CD33 were expressed. Again, marker combinations of mCD33/cCD13 and mCD13 or mCD33/cCD11, respectively, represent a leukemia-related feature, suitable for tracing single leukemia cells in double immunofluorescence. Acute leukemia defined by the coexpression on most blast cells of antigens classically attributed to different lineages (referred as atypical/aberrant marker combinations) remains a rare event. We isolated a series of 27 (12%) such cases of 225 acute leukemia patients whose cells were immunophenotyped at diagnosis. Myeloid markers were present in T-ALL of two cases, T and B markers were coexpressed in 13 cases, markers of B and myeloid lineage were associated in one case, and T cell and myeloid antigens were found in 10 AML cases; in one AML case (M3 according to FAB classification) an aberrant nuclear coexpression of TdT was observed. In one case of the last group an interesting antigen combination of CD4/CD34 present in AML with monocytic differentiation was observed. When 5 patients with leukemia-associated (aberrant) markers were again analyzed at relapse, the relevant antigen combinations were retained in all of them. In summary, 44 of 50 cases (88%) from our acute leukemia series studied for leukemia-associated antigen combination, both with surface membrane and cytoplasmic marker combinations and those with aberrant markers coexpression allow the detection of minimal residual disease.

Potentiation of cis-DDP and pyridoxal effect on isolated DNA during simultaneous application.

Pyridoxal and cis-diamminedichloroplatinum (II) (cis-DDP) have a different mode of interaction with DNA. Cis-DDP caused extensive transforming inactivation of pBR322 DNA but did not form strand breaks in DNA molecules. On the other hand, pyridoxal formed frank strand breaks in the DNA chains and decreased DNA transforming activity. A higher level of DNA transforming inactivation was obtained during simultaneous application of both compounds than would be an additive effect of these compounds applied to DNA independently. This synergistic effect can be ascribed to the introduction of thermolabile sites by the combined action of cis-DDP and pyridoxal. These lesions were converted by heat-treatment to DNA strand breaks and detected electrophoretically. Using two different methods, cis-DDP and pyridoxal, during simultaneous application, interacted with DNA with higher efficiency.

In vitro antiproliferative effect of interferon alpha in solid tumors: a potential predictive test.

An in vitro test for the antiproliferative effect of human leukocyte interferon (IFN-alpha) was performed in primary cultures of tumor cells obtained from 32 patients with either malignant melanoma (13), renal carcinoma (4) or bladder carcinoma (15). Our results demonstrated activity of IFN in all three groups of solid tumors. However, appreciable differences in sensitivity to antiproliferative effect of IFN between individual tumors of the same type were found. The potential of this antiproliferative test for prediction of treatment response in IFN-therapy is discussed.

Humoral immunity in patients with carcinoma of the uterine cervix--effects of radiation therapy.

The parameters of nonspecific immunity-serum immunoglobulins and immune complexes--were evaluated in patients with carcinoma of the uterine cervix (Stages IIB and IIIB) prior to, during and immediately after pelvic irradiation. In untreated patients, significantly elevated circulating IgA was found only in patients in Stage IIIB; serum IgG and IgM in both groups did not differ from control values. The level of circulating immune complexes in both groups was higher than in controls. Radiotherapy did not affect significantly any of the parameters examined; only the percentage of patients with elevated concentrations of IgA and IgG decreased during the treatment. These results showed that fractionated pelvic irradiation did not affect B cell function, these cells being more radioresistant than other lymphocyte subpopulations.