Flow cytometric determination of leukemia-associated marker combinations for the study of minimal residual disease.

To study the minimal residual disease in acute leukemia patients we used some marker combinations related either to the simultaneous surface membrane and cytoplasmic marker expression, or to the expression of atypical marker combinations, that are absent or extremely rare in normal hematopoietic cells. We investigated to which extent flow cytometric analysis of leukemia-associated marker combination may contribute to sensitive follow-up in patients with acute leukemia. For this purpose dilution experiments were performed, in which artificial mixtures of normal peripheral blood lymphocytes and leukemia cells from a patient with leukemia-associated phenotype were prepared and analyzed for double positive cells. Our results showed that the sensitivity of double color immunofluorescence assay was 3 in 10(4) cells. Sequential studies of residual disease were evaluated in five acute leukemia patients with leukemia-associated markers combinations at diagnosis. In three of them morphologic relapse was preceded by the immunologic detection of small amounts of leukemia cells, while in two other cases, in which no double positive cells for leukemia-associated markers were found, patients are still in hematologic remission. This approach to the study of minimal residual disease could be valuable in monitoring the efficiency of chemotherapy, as well as in evaluating the quality control of bone marrow before autografting. Furthermore, flow cytometric approach can efficiently complete other methods, which are used for more exact definition of remission in acute leukemia patients.

Soluble interleukin-2 receptor in serum of patients with esophageal carcinoma.

We measured serum levels of sIL-2R in 28 patients with squamous cell esophageal carcinoma and in 12 controls. Sera for presence of sIL-2R were examined using DAKO Interleukin-2-receptor (CD25) ELISA. The samples were analyzed for CEA and SCC by using IMx System (Abbot). The mean values of sIL-2R in esophageal carcinoma group were significantly increased in comparison with control group. Our results suggest that sIL-2R may be useful in monitoring and prognosis estimation of disease in esophageal carcinoma and could be a valuable addition to the current spectrum of cancer markers.

The value of bone marrow biopsy in the prognosis of hairy cell leukemia (HCL).

Histological findings in the bone marrow and their changes in the course of the disease and therapy were evaluated with respect to the prognosis in a group of 32 patients with hairy cell leukemia (HCL). Two types of bone marrow infiltration by hairy cells (HCs), the diffuse type and the interstitial type, respectively were found. The diffuse type of infiltration and the minimal or absent residual hematopoiesis (RH) at presentation were found with statistical significance to be unfavorable prognostic findings when compared with interstitial infiltration and persisting RH (p < 0.01). Reticulin fibrosis is a characteristic finding in HCL and was found in all but 2 patients.

Correlation of cytosolic concentrations of ER, PS2, Cath-D, TPS, TK and cAMP in primary breast carcinomas.

In a group of 391 patients with primary breast cancer the cytosolic concentrations of ER, PS2, Cath-D, TPS, TK and cAMP were determined. PS2 production was found to be significantly dependent not only on ER but also on cAMP. A similar dependence on ER and cAMP production was found also in Cath-D. When the production of these prognostic factors was correlated with the occurrence of metastases to axillary lymph nodes, three prognostically unfavorable groups of primary breast carcinomas were revealed. The first group was represented by tumors with negative PS2 values (< or = 2.5 ng/mg) and elevated TPS values (> or = 4.0 kU/mg). The second group comprised prognostically very unfavorable tumors with moderately to highly elevated PS2 values and positive Cath-D values (> or = 35 pmol/mg), or positive TPS values (> or = 8.0 kU/mg). The third group was constituted by tumors with elevated TK (> or = 5.0 U/mg) and ER values (> or = 40 fmol/mg). The possible role of PS2 in the metastasizing of primary breast carcinomas is discussed.

Lipoprotein profile in breast cancer women: effect of tamoxifen treatment.

Electrophoretic lipoprotein analyses were performed in 51 patients on tamoxifen and compared with those obtained from 33 newly diagnosed breast cancer patients and with data from the group of healthy women. A statistically significant lower rate of dyslipoproteinemia has been demonstrated in tamoxifen group in comparison with untreated patients; 23.5% vs. 54.5% (p < 0.01). Comparing the results of the latter group with normal subjects there was a significantly higher frequency of dyslipoproteinemia in untreated patients, 54.5% vs. 28.9% (p < 0.05). Our findings confirmed an estrogen-like influence of tamoxifen on lipoprotein profile in postmenopausal breast cancer patients.

The contribution of ultrasonography to the differential diagnosis of breast cancer.

Overall consecutive breast abnormalities (259 carcinomas, 1820 benign) examined with breast ultrasonography (US) are reported. US sensitivity, specificity and positive predictive value were 67.6, 97.7 and 81.0%, resp. (the corresponding values were 57.9, 97.9 and 77.2% for palpation, 79.9, 93.5 and 73.7% for mammography, and 97.6, 92.6 and 87.6% for cytology). US sensitivity was unaffected by age, whereas it was strongly influenced by tumor size (pT1-76.1%; pT2-87.1%) and histologic type (intraductal-7.4%; invasive ductal/lobular-83.4%; invasive special types-64.1%). The features of the lesions at US were significantly associated with cancer (irregular margins, posterior acoustic shadowing) or benign lesions (anechoic structure, lateral shadowing, posterior acoustic enhancement) but had a limited diagnostic accuracy. Overall, US visualized 174 (benign-24, suspicious-150) of 188 palpable, and 32 (benign-7, suspicious-25) of 71 nonpalpable cancers. US contribution was determinant to final diagnosis in 4 of 7 cancers, missed at palpation and mammography, but was at least partially responsible for 8 unnecessary biopsies of benign lesions. A negative/benign US report contributed to avoid unnecessary biopsy in 71 suspicious cases at palpation/mammography. Routine US examination of clinical/mammographic abnormalities is recommended for the advantages of US-guided aspiration and to reduce the frequency of unnecessary biopsies.

The role of cisplatin in chemotherapy of advanced breast cancer.

Cisplatin containing regimens as first-line, second-line or as a third-line chemotherapy were administered in 26 and 36 patients, respectively. The overall response rate in patients on first-line chemotherapy was 53.9%, in patients on second or third-line chemotherapy 30.6%. The differences both in overall and disease-free survival between patients on first-line and on second/third-line chemotherapy were statistically significant in favor of women treated with first-line chemotherapy (p = 0.05). Hematologic and nonhematologic toxicities were mild to moderate and were more pronounced in patients on second and third-line chemotherapy. The overall response rate, DFS and OS were significantly better and longer in the group of patients treated with "bolus" CDDP in comparison to the group of patients treated with CVI CDDP. Our results confirm the activity of cisplatin-containing regimens (mainly CAP schedules) in patients with advanced breast cancer not only as a first-line therapy but also in heavily pretreated patients by chemotherapy and/or radiation therapy and endocrine manipulation.

Prognostic factors in bilateral breast cancer.

Twenty-nine bilateral breast carcinoma patients were analyzed retrospectively. The incidence of bilateral carcinoma of the breast among unilateral breast cancer patients was approximately 2.4%. Median age was 46 years at the time of first cancer diagnosis (range 26-69 years). The majority of the lesions were invasive ductal carcinoma (86%). Of 58 tumors, 10 were staged as Stage I (17%), 30 as Stage II (52%), 8 as Stage III (14%) and 10 as Stage IV (17%). Patients were treated with various combinations of surgery, radiation treatment and chemotherapy. Of 29 patients with bilateral breast cancer, 5 presented with simultaneous bilateral disease (17%), 7 (24%) with synchronous tumors whereas 17 (59%) developed asynchronous tumors. The mean interval between two cancers was 2.6 +/- 0.6 years. Overall survival was 4.8 +/- 0.7 years and overall 5-year actuarial survival was calculated to be 51%. Age, menopausal status and tumor size at the time of initial cancer correlated with the time interval between two cancers. Age, tumor size and nodal status at the time of initial cancer and the time interval between two cancers correlated with the overall survival.

Non-IPSID small intestinal lymphoma: evidence for disseminated disease at presentation.

During the period 1984-1989 the authors have observed 20 patients with non-IPSID small intestinal lymphomas, 11 males and 9 females. In 11 patients the first symptoms were abdominal cramps requiring laparotomy, in 4 ileus, and in 5 perforation with peritonitis. Resection of the involved part of the intestine was performed in 17 patients. Lymphoma tissue was present in 4 of 5 retrogradely examined resection lines on macroscopically normal small intestine. According to Working Formulation, 3 patients had low grade, 3 intermediate grade and 14 high grade histology. Affection of extra intestinal/mesenteric structures was found in 18 of 20 patients, with a total of 36 other lymphoma localizations. 8 of 20 had affection of the nasopharynx and/or Waldeyer's ring. According to the Crowther's classification 55% of patients were in Stage IV, 35% in Stage III and 10% in Stage Ib. All patients were treated with chemotherapy, 13 with ProMACE regimen and 7 with CHOP-type regimens. Ten of twenty patients are alive and in complete remission for over 5 years (7 of 11 of Stage IV and 3 of 9 of Stage Ib/III; 8 of 14 with high grade and 2 of 6 with intermediate/low grade histology). Our results point to the fact that in non-IPSID lymphoma of the small intestine, lymphoma involvement of the intestinal wall might be present beyond obvious lymphoma lesions. Most patients with apparently primary small intestinal lymphoma have a widespread disease. Thus, local forms of treatment such as surgery, and/or radiotherapy can not be expected to be curative in the majority of patients. Data from this study suggest that following initial surgery the chemotherapy is the treatment of choice for these patients.

Modulatory effect of Dalton's lymphoma cells on the production of reactive nitrogen intermediates, interleukin-1 and tumor necrosis factor by murine peritoneal macrophages.

The effect of Dalton's lymphoma (DL) cells on the production of reactive nitrogen intermediates (RNI), interleukin-1 (IL-1) and tumor necrosis factor (TNF) by murine peritoneal macrophages was investigated. Treatment of macrophages with LPS resulted in significant enhancement in the production of RNI, soluble and membrane-associated IL-1 and TNF. A significant inhibition in the production of RNI and IL-1 was observed when the macrophages were coincubated with DL cells whereas TNF production was enhanced. Incubation of macrophages in the presence of DL cells alone even in the absence of LPS could induce production of TNF level comparable to that of LPS-treated macrophages. Paraformaldehyde-fixed DL cells and DL cell conditioned medium could also inhibit RNI production by macrophages. On the other hand pre-exposure of macrophages to viable DL cells or DL cell-conditioned medium prior to treatment with LPS could prime the macrophages for enhanced production of RNI. This study indicates that the regulatory effect of lymphoma cells on macrophage activation may have relevance of the host defence against tumors.

Risk factors for thyroid cancer.

Case-control study comprised 100 histologically verified thyroid cancer patients (23 men, 77 women) and 100 hospital controls matched with cases by sex, age and place of residence. Various risk factors were studied to determine whether they were associated with the occurrence of thyroid cancer. According to the conditional logistic regression analysis, 6 were significantly related to the disease: Cigarette smoking (RR = 7.12 95% CI = 1.53-32.99), family history of any malignant tumors (RR = 5.84 95% CI = 1.76-19.44), history of goiter or thyroid nodules (RR = 27.69 95% CI = 3.11-246.14), long-term occupational exposure to chemicals (RR = 10.07 95% CI = 3.85-26.35), history of second primary tumors (RR = 15.49 95% CI = 3.46-69.30), and diagnostic X-rays exposure (RR = 7.56 95% CI = 2.85-20.07).

Characterization of morphological and histochemical changes induced by overexpression of P-glycoprotein in mouse leukemic cell line L1210.

Vincristine sensitive (L1210) and resistant (L1210/VCR) L1210 mouse leukemia cells were studied from morphological and histochemical point of view. The morphological and histochemical findings reflected differences in membrane structure and in physiological state of sensitive and resistant cells. Numerous villous projections and cytoplasmic protrusions of the cell surface as well as higher activity of membrane enzymes (5'-nucleotidase, ATPase, alkaline phosphatase) were found in vincristine resistant cells. It is assumed that in resistant cells these differences are connected with overexpression of membrane P-glycoprotein. Moreover, in resistant cells more condensed mitochondria were found after their exposure to vincristine. This finding can reflect a higher activity of these organelles in conditions when activity of P-glycoprotein is manifested and is in agreement with increased rate of oxygen consumption by resistant cells from 2.5 +/- 0.3 to 3.3 +/- 0.2 microliter/min.10(6) cells induced by vincristine.

Myeloid activation antigen CD66/67: immunochemical and immunocytometric characterization with the Vth workshop monoclonal antibodies.

Flow cytometric and immunochemical studies performed with the CD66/67 panel antibodies from the Vth International Workshop on Leukocyte Antigens allowed to subdivide these antibodies into five groups, according to the molecular weights of polypeptides recognized by these antibodies on pooled healthy donor granulocytes. Although some monoclonal antibodies recognized either 95-110 kDa, or 160-180 kDa polypeptides, majority of the examined antibodies reacted with epitopes shared by both 95-110 kDa and 160-180 kDa polypeptides. Differential TPA-induced modulation of recognized antigens was observed with HL-60 and U-937 leukemia/lymphoma cell lines.

Association among an autocrine parameter (EGF-R) and endocrine parameters (ER and PR) in locoregional breast cancer.

Sixty-three locoregional breast cancer biopsies were examined for association among epidermal growth factor receptors (as parameter of autocrine growth control), and estrogen and progesterone receptors (as endocrine parameters of estrogen responsiveness). In the tumor group with similar steroid receptor levels, low (0-50 fmol/mg), medium (50-100 fmol/mg) and high (above 100 fmol/mg), an inverse quantitative correlation between epidermal growth factor receptors and progesterone receptors was obtained (p < 0.05). In the tumor groups where estrogen receptor levels were higher or lower than progesterone receptor levels, epidermal growth factor receptors were correlated neither with estrogen nor with progesterone receptors (p > 0.05). It seemed that inverse correlation between endocrine and autocrine parameters obtained in previous studies might not be a common behaviour of all breast tumors.

Lack of expression of adhesion molecules on leukemic cells: possible pathogenetic factor in blood malignancies.

In 68 patients with different leukemias the expression of the following adhesion molecules was examined: CD11a, CD18, CD54, CD44, CD58, and CD59. While in normal individuals all these molecules are broadly expressed on leukocytes, in patients with leukemias the following deviations were observed: (a) at least one of the examined molecules was missing in 64/68 cases (94%); in 12/68 cases (18%) both molecules LFA-1 and ICAM-1 were missing, in 37/68 (54%) either LFA-1 or ICAM-1, and in 15/68 cases (22%) adhesion molecules other than LFA-1 or ICAM-1 were missing; (b) the expression of CD11a/CE18, CD58, CD59 on leukemic cells was heterogeneous, without any clear correlation to the subclass of leukemia; (c) in the majority of cases, CD54 (45/68; 66%) and CD44 (36/68; 53%) were missing, however showing a tendency of expression on leukemic cells with more mature immunophenotype.

Cytotoxicity versus transforming activity in chemically exposed Syrian hamster embryo cells.

A series of experiments was conducted to determine the dose response relationship with respect to both cytotoxicity and morphological transformation in Syrian hamster embryo cells exposed to different chemicals. The effects of the following model chemical carcinogen/mutagens were investigated: 3-methyl cholanthrene (3-MC), benzo(a)pyrene (B(a)P), N-methyl-N-nitro-N-nitroso-guanidine (MNNG), N-methyl-N-nitrosourea (MNU), the noncarcinogenic weak mutagen methyl methanesulfonate (MMS) and nonmutagenic pesticides Supercypermethrin EC, VUCHT 524, and Dual. The results showed that all carcinogen/mutagens, i.e. 3-MC, B(a)P, MNNG, and MNU increased the number of morphological transformations on the maximum level at concentrations < TD50-TD70 (TD = toxic dose) and remained more or less on the same level at higher concentrations. Similar effect was observed in embryo cells treated with nonmutagenic Supercypermethrin EC. Very low concentrations of nonmutagenic pesticide VUCHT 524 stimulated proliferation of cells and at the same time induced the maximum level of morphological transformations. MMS and Dual did not induce morphological transformation of embryo cells at all. Induction of morphological transformation in embryo cells is evidently independent of the activity of a chemical to induce gene mutations and seems to be a valuable assay for studying the carcinogenic effects of lower doses of suspicious chemicals.

3-Aminobenzamide delays rejoining of DNA strand breaks in gamma-irradiated lymphocytes from patients with breast cancer and not cervical cancer.

An inhibitor of poly(ADP-ribose) polymerase is 3-aminobenzamide (3AB), treatment by which normally results in the decreased levels of rejoining of DNA single strand breaks. We have studied the effect of 3AB in gamma-irradiation induced damage and the subsequent repair of DNA along with the corresponding changes in poly (ADP-ribose) polymerase activity in lymphocytes of patients with the cancer of breast and uterine cervix. The presence or absence of 3AB prior to gamma-irradiation did not influence the extent of DNA damage. On the other hand, single strand breaks rejoining in breast cancer cases was found to be slower as compared to that of cervical cancer cases and normal individuals. The relative ADP-ribosylation in the presence of 3AB was much lower in the breast cancer cases as compared to normal but there was no significant change in the cervical cancer cases. Overall the maximum relative ADP-ribosylation was slower in the presence of 3AB. This suggests that the lower activity of poly (ADP-ribose) polymerase in breast cancer cases might delay the first phase of single strand breaks (SSBs) rejoining.

The role of alpha-difluoromethyl ornithine as an adjuvant to immunotherapy in mice bearing transplantable tumors.

alpha-Difluoromethyl ornithine (DFMO), an inhibitor of ornithine decarboxylase (ODC), depletes cellular polyamine levels which is significantly correlated to neoplastic transformation. DFMO was used to modulate the growth rate of Ehrlich ascites carcinoma in mice and attempts were made to use DFMO as an adjuvant to immunotherapy. The therapeutic efficacy was estimated by measurement of polyamine levels of blood and ODC activity. Significant changes were also observed in the survival of the treated groups of animals, the implications of which have been discussed.

Human larynx carcinoma cells resistant to cis-diamminedichloroplatinum(II): mechanisms involved in the resistance.

The aim of this study was to characterize two cis-diamminedichloroplatinum(II) (CDDP) resistant cell lines established from human larynx carcinoma HEp2 cells through repeated treatments with increased CDDP concentrations. CK2 cells obtained by continuous treatments were more resistant to CDDP than CA3 cells obtained by acute treatments. The examination of growth characteristics showed that both CDDP resistant cells had doubling times identical to that of the parental cells, but had lower plating efficiency. The possible involvement of glutathione (GSH), glutathione transferases (GST), metallothioneins, P-glycoprotein and drug accumulation in CDDP resistance was examined. Glutathione contents were elevated in both CDDP resistant lines. However, neither GSH nor GST were involved in CDDP resistance. This was demonstrated by simultaneous incubation of parental and CDDP resistant cells with CDDP and specific inhibitors of GSH and GST alpha and pi (buthionine sulfoximine and ethacrinic acid). Similarly, verapamil, an inhibitor of P-glycoprotein, did not influence the sensitivity of parental and resistant cells to CDDP. As compared to the parental cells, CK2 cells became resistant and CA3 cells became sensitive to cadmium, indicating increased level of metallothioneins in CK2 cells, and reduced level in CA3 cells. Measurements of platinum contents in parental and CDDP resistant cells after 1, 3 and 6 hours exposure to 70 mumol CDDP showed reduction in platinum accumulation after each exposure time in CK2 cells, and after 6 hours exposure in CA3 cells. This study identified decreased platinum accumulation as an important mechanism of CDDP resistance in human larynx carcinoma cells.

Human larynx carcinoma cells resistant to cis-diamminedichloroplatinum(II): cross-resistance patterns.

In our previous study we developed two lines of human larynx carcinoma HEp2 cells resistant to cis-diamminedichloroplatinum(II) (CDDP) due to acute (CA3 cells), or continuous (CK2 cells) treatments with increased CDDP concentrations. CA3 line was 2.3-fold resistant, and CK2 line 3.7-fold resistant to CDDP as compared to the parental cells. In this study the sensitivity of CDDP resistant and parental cells to several antitumor drugs was examined using clonogenic survival assay. CK2 cells were 1.7-fold more resistant to vincristine than parental cells, while CA3 cells exhibited resistance to vincristine only at higher concentrations of this drug. Verapamil, an inhibitor of P-glycoprotein, was not able to reverse the resistance of CK2 cells to vincristine, indicating that this cross-resistance did not involve P-glycoprotein. As compared to the parental cells, CK2 cells were 2.1-fold resistant, and CA3 cells 1.7-fold resistant to mitomycin C. CK2 cells were 2-fold more resistant to 5-fluorouracil than the parental cells, while CA3 cells did not exhibit any change in the sensitivity to this drug. CA3 cells were 0.4-fold sensitive to epoxide and 0.53-fold sensitive to doxorubicin, while CK2 cells had the same sensitivity to these drugs as the parental cells. Our results showed that treatment schedule determines the response of CDDP resistant human larynx carcinoma cells to additional drugs, suggesting the complexity of the judicious choice of the drugs in the combined modality treatments of cancer.

Evaluation of chemopreventive effects of betel leaf on the genotoxicity of pan masala.

The antigenotoxic effect of the aqueous extract of betel leaf (BL-ext.) against the pan masala was tested with the help of cytogenetic endpoints like chromosome aberration (CA) and sister chromatid exchange (SCE) utilizing Chinese hamster ovary (CHO) cells. Compared to the cultures treated with aqueous extract of pan masala alone, a reduction in CA and SCE frequencies in CHO cells was observed following a combined treatment with pan masala (with or without tobacco) extract and BL-ext. The protective effect of BL-ext. against the genomic damage caused by pan masala was statistically significant only after treating the cells for a longer period.

Effect of immobilization stress on tumor growth in mice.

The effect of immobilization stress applied daily for 2 h on the development of MF-RSV tumor graft in mice and on survival of the hosts was studied. While a single acute stress applied to mice simultaneously with a transfer of MF-RSV cells stimulated antitumor defense, chronic stress applied both before and/or after tumor cell implantation accelerated the death of recipients. The shortest survival time was found in mice which were stressed for 3 weeks before implantation and on, until their death. Heat shock applied to MF-RSV tumor cells before implantation significantly reduced their growth ability in the hosts.

Effect of fibrosarcoma induction on copper and ceruloplasmin concentration in different organs of the host.

The copper content and ceruloplasmin activity were determined in mice bearing benzo(a)pyrene induced fibrosarcoma. The copper level and ceruloplasmin activity in different organs of fibrosarcomatous mice varied when compared to their controls. Significant changes in copper and ceruloplasmin concentration were observed in the liver and tumor tissue of host mice bearing fibrosarcoma compared to controls. Disturbed copper metabolism at the hepatic level may account for the hypercupremia observed during malignancy.

Hemopoietic cell differentiation antigens (CD system) 1993. Minireview.

Subcutaneous interleukin-2 in combination with vinblastine for metastatic renal cancer: cytolytic activity of peripheral blood lymphocytes.

Eight patients with progressive metastatic renal cell carcinoma were selected for one course of subcutaneous recombinant interleukin-2 (IL-2) plus vinblastine (VBL) treatment lasting for seven weeks. Seven of the eight patients were evaluable for response, eight for toxicity. Peripheral blood lymphocytes (PBL) from the evaluable patients were isolated and frozen prior to, during, and after the treatment courses; kinetics of their cytolytic activity was assessed and compared under standard conditions in 51Cr microcytotoxicity assay with natural killer (NK)-sensitive and NK-resistant human tumor targets. Among the evaluable patients treated, there was 1 partial responder (10+ months, regressions occurred in lung, retroperitoneal lymph nodes and adrenal metastases) and 3 patients achieved a stable disease (10+, 10+, 5+ months). Systemic toxicity was mild to moderate with treatment-limiting adverse effects in one patient (severe thrombocytopenia, grade IV). In the IL-2-treated patients, the cytolytic activity of PBL directed against NK-sensitive targets rapidly decreased during the first week of IL-2 treatment, approaching the negative values on day 10. Then the cytolytic activity was slowly increasing and reached its maximum within another two weeks. Afterwards, the cytolytic activity of PBL was again decreasing and the approximate values of the initial cytolysis were reached after 6-8 weeks. In contrast, with NK-resistant targets such characteristic kinetics of PBL cytolytic activity was not observed. The kinetics of PBL-mediated cytolysis was similar in IL-2-responders and non-responders, so that no correlation of in vivo and in vitro effects of subcutaneous IL-2 and VBL treatment could be established.

Application of epirubicin containing albumin microspheres in the experimental therapy of breast cancer.

In the experiment we attempted to use albumin microspheres as carriers of cytostatics to acquire a higher drug concentration in the tumor for a longer time. It was expected that the efficiency of a treatment with a microsphere-entrapped epirubicin would be better than a treatment with microsphere-free drug. Albumin microspheres containing epirubicin were prepared by ultrasonic homogenization in oil and heat denaturation. In vitro experiments and the test for biological activity on human breast carcinoma cells implanted into Nu/Nu mice were carried out. Growth of tumors in the group of mice treated with microsphere-entrapped epirubicin was inhibited more effectively in comparison with growth of tumors in the group treated with microsphere-free drug. These results suggest that the change of the drug form can enhance antitumor effect of epirubicin and that albumin microspheres are suitable drug carriers.

HPLC-ED determination of catecholamines and their metabolites in urine.

High performance liquid chromatography (HPLC) with electrochemical detection (ED) was applied for the monitoring of catecholamines (epinephrine - E, norepinephrine - NE, dopamine - DA) and their main metabolites (homovanillic acid - HVA, vanillylmandelic acid - VMA) in urine samples of patients with the pheochromocytoma diagnosis. Both amperometric and colorimetric detectors were tested for the clinical applications and the results were compared. The optimization of separation conditions was worked out, especially the effects of mobile phase compositions (pH, ionic strength, organic modifier and ion-pair agent concentrations). Dependences of capacity ratio values k' on all optimized components of the mobile phases were evaluated. Chromatographic conditions were optimized for the suitable chromatographic resolution (Rij > 1.25). Extraction recoveries for liquid-liquid and solid-phase extractions have been worked-out. Detection limits for catecholamines in urine were in the range of 0.3-0.6 ng/ml and 10-20 ng/ml for HVA, VMA using coulometric detector.

Evaluation of CA 15-3 tumor marker in the diagnosis of breast cancer. A pilot study.

Single determinations of serum CA 15-3 levels were performed by sandwich enzyme immunoassay in 160 women: 77 patients with nonmalignant breast tumors (64 had fibrocystic disease, 11 had fibroadenoma, 2 had intraductal papilloma) and 83 patients with primary breast cancer prior to any treatment; the cut-off limit was established at 30 U/ml. The overall diagnostic sensitivity and specificity of the CA 15-3 test was 19.3% and 94.8%, respectively. The positive and negative predictive values were 80.0% and 52.1%. The mean CA 15-3 value was significantly lower in patients with benign breast tumors as compared with the breast cancer group: 16.8 +/- 8.2 vs. 23.9 +/- 20.9 U/ml (p < 0.01) as well as the percentage of positive results of the test: 5.2% vs. 19.3% (p < 0.02). Serum CA 15-3 level in breast cancer patients correlated with: (1) clinical stage: a higher percentage of positive results was observed in patients with more advanced cancer: Stage I-0%, Stage II-10.6%, Stage III-29.6%, and Stage IV-100.0% according to UICC classification; the comparison of breast cancer patients with early stage of disease (I+II) and those with more advanced cancer (III+IV) revealed statistically significant (p < 0.01) difference in the mean serum CA 15-3 value (19.7 +/- 12.8 vs. 31.5 +/- 29.2 U/ml) as well as in the percentage of positive results (9.4% vs. 36.7%, p < 0.01); (2) the histological grading according to Bloom and Richardson: 5.41% of positivity was observed in low and intermediate grade cancers (I+II) vs. 66.7% in grade III (p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Red cell ferritin and iron stores in chronic granulocytic leukemia.

Basic red cell ferritin was investigated in 28 patients with different phases of chronic granulocytic leukemia (CGL). Red cell ferritin was significantly decreased in remission after busulphan treatment and significantly elevated in the blast crisis as compared to healthy controls. Bone marrow stainable iron was decreased or absent in 86% of patients in the initial phase at the time of diagnosis and in 92% of those in remission. Red cell ferritin correlated with serum ferritin, however, serum ferritin level remained above normal range during all phases of the disease. A negative correlation between red cell ferritin and hemoglobin (Hb) (r = -0.605, p < 0.001) suggested that red cell ferritin level reflected the rate of iron utilization for heme synthesis. Decreased red cell iron stores observed in the remission may be explained by regression of dyserythropoiesis and by restoration of normal Hb synthesis after busulphan treatment. A progressive dyserythropoiesis in the blast crisis may lead to an increased red cell ferritin level.

Liver calcium homeostasis modification by iron: a probable factor in its carcinogenesis.

Low molecular weight iron complexes, ferric lactate and ferric-ATP complex, induce an important increase of Ca(2+)-uptake by liver. The activity of ferric lactate increased by the presence of sodium ATP, and the steady high effect of ferric-ATP complex appear to indicate that ATP might play an important role in the in vivo formation of low molecular weight iron complexes that can induce the modification of the hepatocytes calcium homeostasis, the event that might be one of the factors triggering the malignant transformation.