Soluble IL-6 and TNF receptors release by polymorphonuclear and mononuclear cells in cancer patients.

Immune response of the host to tumor is regulated by soluble mediators, such as cytokines. Data including the release of soluble mediators by polymorphonuclear cells (PMNs) are controversial. In the present investigation we studied the ability of spontaneous and LPS-stimulated PMNs derived from the patients with oral cavity cancer to the release of soluble IL-6 receptor (sIL-6R) and soluble TNF receptors (sTNF-Rs). Obtained results were compared to the release of sIL-6R and sTNFRs by peripheral blood mononuclear cells (PBMC) as well as the serum levels. The culture supernatants of spontaneous and LPS-stimulated PMNs derived from the patients with oral cavity cancer contained unchanged concentrations of sIL-6R and higher concentrations of sTNFRp55 and sTNFRp75 in comparison with the control. The amounts of sIL-6R and sTNF-Rs released by PBMC were higher than those released by PMNs in both control and patient groups. Demonstrated results indicate that PMN contribution in the immune response is mediated by TNF-alpha, via the release of sTNFRp75 mainly. Profile measurement of the soluble cytokine receptors in the culture supernatants of PMNs and PBMC and the serum levels may be useful for estimation of the actual immunity in patients with oral cavity cancer.

Radiosensitivity of human lymphocytes in vitro correlates more with proliferative ability of cells than with the incidence of radiation-induced damages of the genome.

The purpose of this study was the estimate the extent of individual variability in radioresponse of human lymphocytes in vitro and to establish the reasons of variability. Individual variability in radiation-response was evaluated using the cytochalasin B micronucleus test among 82 healthy individuals (36 men and 46 women), of mean age 38 (range 30-48). Blood samples were irradiated with gamma (60Co) rays at a dose of 2 Gy in vitro. The yield of radiation-induced micronuclei (MN), cytochalasin blocked proliferation index (CBPI), fraction of micronucleated binucleate (BN) cells and mean incidence of MN per micronucleated BN cells at a sampling time of 72 hours were scored. Our results brought out a significant effect of gender on the level of spontaneously occurring micronuclei, the lack of statistical differences between gender in the yield of radiation-induced micronuclei and marked variability in radiation response among individuals. Likelihood of expressing hypersensitivity was correlated with ability of cells to proliferate in vitro (beta = 0.41, p < 0.000) more than with the incidence of radiation-induced micronuclei per micronucleated cell (beta = 0.20, p < 0.000).

Crosslinking of cellular DNA by nitracrine and furocoumarin derivatives.

The anticancer drug, nitracrine, a 1-nitro-9-aminoalkyl derivative of acridine exhibits potent cytotoxic effects which are due to its metabolic activation, followed by covalent binding to macromolecules--DNA being the target for the drug. The renaturable fraction of DNA from L-1210 cells pretreated with nitracrine is assayed by means of ethidium bromide fluorescence assay and chromatography on hydroxyapatite column. The effect of the drug was compared with furocoumarins of different DNA crosslinking potencies. The existence of crosslinks in DNA upon incubation of cells with nitracrine (1-4 microM) have been confirmed with two different methods under the conditions where 8-methoxypsoralen, a classic crosslinking agent induced the renaturation. The DNA preparation isolated from the drug pretreated cells exhibited decreased transcriptional template activity with E. coli DNA-dependent RNA polymerase.

The influence of surgery and anesthesia on lymphocyte functions in breast cancer patients: in vitro effects of indomethacin.

Surgical trauma and anesthesia may lead to the postoperative immunosuppression, the exact mechanism of which is still unresolved. Among various factors, the role of prostaglandine PGE2-mediated suppression was also proposed. We investigated the influence of surgery and two anesthetic regimes on lymphoproliferative response (LPR) to PHA and on NK cell activity (MTT) in breast cancer patients, as well as the effect of indomethacin, a PGE2 synthesis inhibitor, on these lymphocyte functions in vitro. In 36 previously untreated patients the lymphocyte functions were assayed before, 24 hours and seven days after the surgery. In regard to LPR, three distinct response patterns were observed: a) significant (p < 0.05) increase of initially lowered LPR; b) significant (p < 0.001) decrease of initially normal LPR 24 hours after operation, followed by normalization after seven days; c) no change of initially normal LPR. Indomethacin in vitro significantly (p < 0.05) enhanced the diminished LPR only before surgery, no effect being seen after the operation. The NK cell function was unaffected by surgery regardless the initial level of activity. Indomethacin had no effect on this lymphocyte function. There was no difference between the patient groups submitted to the different anesthetic regimes. In conclusion, our results show that surgical trauma variably affect the lymphocyte functions of cancer patients, the effect not being related to the particular anesthetic regime used. The PGE2-mediated suppression is not likely to be involved in postoperative immune function impairment.

Sequential standard dose mitoxantrone and cytosine arabinoside for newly diagnosed adult acute myeloblastic leukemia.

Twenty one adult patients with previously untreated acute myeloblastic leukemia (AML) were treated with sequential mitoxantrone and standard dose cytosine arabinoside remission induction therapy. The median age was 33 years (range 17-56 years). Complete remission (CR) was achieved in 80% (17/21 cases) and 76% (16/21 cases) achieved CR after one course of induction therapy. The median duration of disease free survival was 9 months with an actuarial disease free survival of 22% at 43 months. The non-hematological toxicity was acceptable. We conclude that sequential mitoxantrone and cytosine arabinoside combination therapy is an effective antileukemic regimen which produces high CR rates in previously untreated adult patients with AML.

Comparative estimation of cure rates for supraglottic and glottic cancer in radiotherapy.

There are some clinical evidences, that the same types of tumors originated from neighboring anatomical structures can significantly differ in their response to radiation therapy. Squamous cell cancer of supraglottis and glottis could be good examples of this phenomenon. The purpose of the study was to compare the radiocurability of cancers localized in the upper and medium level of the larynx. From 1985 to the end of 1989, 544 patients with squamous cell cancer of the larynx were treated by radiotherapy alone. There were 388 patients with supraglottic cancer and 156 patients with glottic cancer. The total dose was in the range of 59-74 Gy. The end-point criteria were overall (OS) and disease-free survival (DFS). Generally, 5-year overall and disease-free survival rates were significantly more favorable for glottic cancer patients than for supraglottic cancer (67 and 63% vs. 40 and 36%, respectively). Significant differences in both disease-free and overall survival between supraglottic and glottic cancer in aspect of several analyzed clinical prognostic factors were found for: male sex, age, pattern of tumor growth, clinical performance status, radiation total dose lower than 70 Gy, fraction doses and overall treatment time. In all these prognostic categories 5-year survival rates were lower for supraglottic cancer patients. This tendency disappeared when the treatment results were compared in aspect of tumor stage (T). Tumor cure doses for 50% probability of local control (TCD50) in supraglottic cancer were estimated as: 61 Gy (T(1+2)) and 66 Gy (T3). In glottic cancer the lower TCD50 values of 54.5 Gy (T(1+2)) and 61 Gy (T3) were found in comparable treatment time. The comparative estimation of cure rates (i.e. OS and DFS) of laryngeal cancer treated by radiation alone showed that in aspect of almost all analyzed prognostic factors the greater risk of treatment failure was significantly associated with supraglottic origin.

Cancer departments as a source of resistant bacteria and fungi?

Antimicrobial resistance increases worldwide. Among many factors, such as clonal spread of genes of resistance among and intra species, local epidemiology, nosocomial transmission, also consumption of antimicrobials may be responsible. Cancer departments, mainly in centers treating hematologic malignancies and organizing bone marrow transplantation (BMT) are known to have extensive consumption of either prophylactically or therapeutically administered antibiotics and antifungals. It is worthy to remember, that first strains of quinolone resistant E. coli, vancomycin resistant enterococci and staphylococci and fluconazol-resistant Candida albicans appeared in the patients treated for cancer with antineoplastic chemotherapy, resulting in profound granulocytopenia. Therefore, assessment of risks of antibiotic prophylaxis with quinolones and azoles and extensive use of empiric therapy with glycopeptides and polyenes needs to be considered. Intensive prophylactic strategies should be limited to group of high risk, leukemic patients or BMT recipients.

About the antiemetic effectivity of granisetron in chemotherapy-induced acute emesis: a comparison of results with intravenous and oral dosing.

The paper is devoted to review literature data on intravenous and oral antiemetic effectivity of granisetron (GRAN) a selective 5HT3 antagonist and to determine the optimal dose for the prophylaxis of chemotherapy-induced acute emesis. The drug was put on the market in the injectable form in 1994 and in the oral form in 1995, so a sufficient number of reports have been published for the evaluation. According to the summarized data on 6095 patients treated with intravenous GRAN, on average 66% antiemetic complete response (CR) rate was reached (i.e. no vomiting in the first 24 hours of chemotherapy). Best results were observed with the dose of 40 microg/kg intravenous GRAN, on average 70% CR (range 47-93%) were achieved in 4182 of the 6095 patients with this dose. In 942 patients treated with the mostly applied oral dose of GRAN (1 mg twice daily), on average 61% CR (range 52-82%) were reported. Side effects were weak and transient, mostly headache and constipation were observed. Headache appeared in 13% with the use of 40 microg/kg intravenous dose GRAN and in 18% with 1 mg oral dose twice daily Constipation was observed in 5.3% and 17% with the injection and oral dose, respectively.

FASAY: a simple functional assay in yeast for identification of p53 mutation in tumors.

Alteration of the p53 tumor suppressor gene is the most common genetic defect known to occur in human tumors. Germ-line p53 mutations significantly increase the risk of developing diverse malignancies. FASAY is a simple functional assay for germ-line and somatic mutations in the p53 gene altering the transactivation capability of the p53 protein. The method was successfully used for mutation analysis of p53 in various cell lines, somatic tumor cells and blood cells. In addition, FASAY was also found effective as a tool for basic research of binding of mutant p53 proteins to promoters of different p53 target genes.

Kojic acid--a new leading molecule for a preparation of compounds with an anti-neoplastic potential.

Kojic acid as a molecule of natural origin may serve as template for the synthesis of new biologically active compounds. The synthetic KA (pyranone) derivatives possess various kinds of biological activities which are related by their similarity to flavonoids. The most important property is the antifungal and antineoplastic activity and capability of chelating metals. It is shown that the antineoplastic activity of kojic acid derivatives is based on various mechanisms of action on different levels of cellular metabolism and functions what makes this compound interesting for future investigation as cytotoxic agent.

Symptom pattern and diagnostic work-up of malignancy at first symptom presentation as related to level of care. A retrospective study from the primary health care centre area of Kungsbacka, Sweden.

This retrospective study was aimed to characterize the diagnostic process of cancer with respect to level of care, initial symptoms, and diagnostic procedures. It was based on analysis of medical records of all subjects with colorectal, pulmonary, breast or prostate cancer, reported to the Swedish Cancer Registry during defined periods of time in the community of Kungsbacka with about 46,500 inhabitants. Initial symptoms, diagnostic procedures, outcome of diagnostic procedures, level of care, and doctor's delay were analyzed. Most patients (62-73% for the different cancers studied) first visited a general practitioner for the symptoms which lead to the diagnosis of cancer. The most common initial symptom for colorectal cancer was defecation abnormality, for breast cancer a palpable mass in the breast, for pulmonary cancer cough, and for prostate cancer symptoms of prostatism. There was no difference in doctor's delay between general practitioners and other physicians. Nonspecific blood laboratory tests made little contribution to the diagnosis of cancer. The results indicate that most cancers of the types studied are diagnosed in primary health care and that it is possible to improve the identification of the few malignant cases among the "noise" of benign diseases, both with respect to accuracy and cost-effectiveness. It seems that focused investigations such as fecal occult blood tests and rectoscopy should be more frequently used in patients with gastrointestinal symptoms.

Copper, zinc and superoxide dismutase in precancerous, benign diseases and gastric, colorectal and breast cancer.

The aim of the present study was to assess serum levels of copper and zinc levels and erythrocytes Cu,Zn-SOD activity and to determine probable changes in gastric and colorectal precancerous diseases, benign breast diseases, gastric, colorectal and breast cancer. The study included 165 subjects with cancer, 348 subjects with precancerous (atrophic gastritis, gastric adenoma, colon adenoma, rectal adenoma) and/or benign diseases (weak dysplasia, severe dysplasia, fibroadenoma, cystic disease) and 161 randomly selected healthy controls. Our results suggest that while in gastric and colorectal cancer there were mostly increased copper levels, in breast cancer they were not changed. Zinc levels were weakly decreased in atrophic gastritis, gastric adenoma and breast cancer. There was a strong positive correlation between zinc levels and SOD activity in fibroadenoma and a weak positive correlation in colorectal adenoma and colorectal cancer without any correlation between SOD activity and copper in these groups. In gastric precancerous disease there was a positive correlation between SOD and copper. The results of this study suggest that serum trace element levels and activity of related enzymes might be different in various neoplastic processes. This variation in neoplastic processes might be influenced by other factors that have to be considered in complex relationships between the whole body and neoplastic cells.

Inhibitory effect of Trianthema portulacastrum L. diethylnitroso-amine-induced phenobarbital promoted hepatocarcinogenesis.

The chloroform extract (active components from the exotic weed Trianthema portulacastrum L. of Aizoaceae, showed remarkable effect in the reduction of DENA-induced rat hepatocarcinogenesis. Hepatocarcinogenesis was induced in male Sprague-Dawley rats by a single intraperitonial injection of diethylnitrosoamine (DENA) at a dose of 200 mg/kg body weight. Chloroform extract of T. portulacastrum at a dose of 100 mg/kg/body wt. to the basal medium/per animal once daily was administered following DENA injection. Morphometric evaluation revealed that the fraction reduced the incidence, numerical preponderance, multiplicity and size distribution of visible pre-neoplastic nodules. Further focal lesions showed a reduction of altered liver cell foci/cm2 and a reduction of average focal area. A decrease in the percentage of liver parenchyma occupied by foci all seem to suggest the anticarcinogenic potential of Trianthema portulacastrum in DENA-induced rat hepatocarcinogenesis.

Estradiol receptor and prognostic parameters of human breast cancer.

Estradiol receptors are regarded to predict a likely success of hormonal therapeutic efforts and the prognosis of breast cancer patients. But today its prognostic importance is controversial, discussed as either reflecting intrinsic property of the tumor tissue or better therapeutic accessibility of receptor positive tumors. Moreover, the most important clinical prognosticators--tumor size and axillary lymph node involvement do not seem to be related to the estradiol receptor status. In our investigation, the length of disease free interval is similar in estradiol receptor positive and negative patients and in all sites of distant metastases, but it is significantly reduced if more than 4 axillary lymph nodes are involved. Post recurrence survival is significantly longer in estradiol receptor positive than negative patients and also in patients treated by tamoxifen containing therapies. Its length is independent of the number of axillary lymph node metastases and the type of distant metastases, with a tendency to be longer in estradiol receptor positive than negative patients. In addition, the overall survival is longer for estradiol receptor positive than negative patients and becomes reduced with more than 4 axillary lymph node metastases. Frequency of deaths in estradiol receptor positive patients is half that of negative subjects. Furthermore, the length of overall survival is independent on the type of distant metastases, with tendency to be longer in estradiol receptor positive than negative patients. Longest overall survival could be observed for estradiol receptor positive patients who got therapy regimens containing tamoxifen. The weak prognostic advantages of estradiol receptor positive patients are interpreted by estradiol receptors as intrinsic parameters of breast cancer tissue characterizing more its biological behavior than therapeutic accessibility.

Enhanced activity of estramustine, vinblastine, etoposide, and suramin in prostate carcinoma.

Following hormonal therapy, few treatment regimens have activity in metastatic prostate cancer. Cytotoxic agents have minimal activity in this disease. However, combinations of cytotoxic agents may be beneficial. The activity of estramustine, vinblastine, etoposide, and suramin on cell growth was evaluated. Prostate specific antigen (PSA) is routinely used as a surrogate marker for disease progression. Many pharmacological agents alter PSA levels independently of their effect on tumor growth, the effect of these agents on PSA secretion was determined. Each agent was evaluated alone and in combination with the other drugs in two prostate cancer cell lines. In LNCaP cells, estramustine and suramin were cytostatic, while vinblastine and etoposide were cytotoxic. Estramustine down-regulated etoposide PSA secretion, while suramin had no effect. The effects of etoposide and vinblastine on PSA secretion were not evaluable. In PC-3 cells, only etoposide was cytotoxic. Tandem combinations were more cytotoxic than single agents in both cell lines. The addition of a third agent to the tandem combination produced less cytotoxicity. In our hands, the best combinations were estramustine/vinblastine, suramin/vinblastine, and suramin/etoposide. These combinations yielded 20-60% higher cytotoxicity than any of the drugs alone.

The surgical treatment of adrenal gland tumors--incidentaloma.

Analysis of 32 patients operated on because of accidentally discovered adrenal tumors "incidentaloma" is presented. In 12 of them there was subclinical hormonal activity, in 9 of them tumors turned out to be pheochromocytoma and 3 of them were cortex adenoma. There were 20 hormonally inactive tumors, in 5 of them there were malignant lesions (4 of the cortex and 1 of the medulla). For evaluation of hormonal activity of adrenal tumors evaluation of chromogranin A and cortisol serum blood level or urine free cortisol level is recommended. For precise localization of the tumor beside USG also CT examination is of use. According to the high percentage of malignant lesions in "incidentaloma" type tumors, surgery treatment without delay is recommended. BAC or DHES in blood serum examinations were not found helpful in preoperative evaluating the lesions as benign or malignant. In case of preoperatively found subclinical hormonal hyperactivity of medulla pharmacological treatment with alpha and beta blockers in surgery preparation is recommended. Lateral extraperitoneal access for adrenalectomy is considered safe and provides good operational view. Laparoscopic procedure because of high percentage of malignant lesions in this group of patients is not justified.

Primary cancer of the fallopian tube with transitional differentiation. Clinical and pathological assessment of 6 cases.

To establish prognosis, histologic appearance and p53 and c-erbB-2 expression in cancer tissue, six cases of primary transitional cancer of the fallopian tube were analyzed. Among 45 patients with the primary cancer of the fallopian tube diagnosed between 1992 and 1997, we found six cases diagnosed previously as solid (undifferentiated) cancer of the tube. p53 protein and c-erbB-2 oncoprotein expression were examined using an avidin-biothinyl-peroxydase complex method. The accumulation of p-53 protein and c-erbB-2 oncoprotein were used as prognostic marker of the transitional cancer of the tube. According to histologic picture all patients were diagnosed for primary cancer of the tube with transitional differentiation. In 4 cases strong positive and in 2 cases moderate positive reaction with antibody against p53 protein was seen for p53 protein. No positive expression of c-erbB-2 oncoprotein in membrane of cancer cells in our cases was detected. Presence p-53 protein in all our 6 cases deny the usefulness of the p53 protein as prognostic marker in primary cancer of the fallopian tube. Lack of expression of c-erbB-2 oncoprotein in membrane of cancer cells is significantly contributed to better prognosis in cases with primary cancer of the tube with transitional differentiation.

The role of stable disease in objective response assessment and its impact on survival in advanced colorectal cancer: is "stable disease" a homogenous response category?

Stable disease is a category which is not included in the evaluation of the overall treatment response rate. In many studies with a response rate below 20%, chemotherapy almost doubles the survival of patients. In the most chemotherapy trials with advanced colorectal cancer patients, about 30-50% had stable disease. Despite belonging to the same category of therapy response, some patients with stable disease have achieved symptom improvement, but some have not. The aim of the study was to investigate whether the stabilization of the disease with clinical benefit is associated with benefit in survival. A total of 99 patients with advanced colorectal cancer were treated with carboplatin (80 mg/m2, day 1-7), 5-FU (750 mg/m2, day 1-5), leucovorin (100 mg/m2, day 1-5) every 4 weeks. After 4 courses, in the case of stable disease (SD), the patients were stratified according to clinical benefit achievement in: Group A--patients with clinical benefit who continued with chemotherapy until 8 cycles or until disease progression; group B--patients without clinical benefit in whom chemotherapy was stopped after 4 cycles. Clinical benefit was a composite of assessment of pain, ECOG performance status, weight and temperature. Clinical benefit required a sustained improvement in at least one parameter without worsening in any other. Of 97 evaluable patients 48 achieved stable disease. Of 22 pts. with SD clinical benefit performance status improvement was recorded in 17, pain relief in 14, improvement in body weight in 14 and temperature disappearance in 8 pts. Of 26 pts. with SD without clinical benefit, 7 were asymptomatic from beginning of the chemotherapy. No difference was detected in the survival between responders and SD clinical benefit pts. (p = 0.24), but there was significant difference between responders and SD pts. without clinical benefit (p = 0.0004). SD clinical benefit pts. had significant difference in survival in comparison to pts. with progressive disease (p = 5.1 x 10(-6)). The results of our study indicate that under category "stable disease" there are two different subpopulations of patients with quite different symptom response to chemotherapy, different time to progression and possible different survival.

Renal cell carcinoma-associated immune impairment that may interfere with the response to cytokine therapy.

This prospective study was carried out to explore cytokine-related immune alterations in 69 renal cell carcinoma patients (RCC) and to look for changes which might potentially serve as a reliable predictors of response to cytokine-based therapy. Interleukin-2 (IL-2), its soluble receptor (sIL-2R) and tumor necrosis factor (TNF-alpha) levels produced in vitro by PHA activated and intact mononuclear cells (PBMC) were determined. Concentrations of IL-2, IL-4, IL-6, sIL-2R, TNF-alpha and CRP were measured in sera. Cytokine level was evaluated by enzyme-linked immunoadsorbent assay (ELISA) and CRP was determined by means of turbidimetric method. All measurements were performed in patients without any prior treatment. PHA activated PBMC of RCC patients were significantly defective in producing IL-2 and TNF-alpha comparing to controls (p < 0.03 and p < 0.001). The difference of sIL-2R was noted in metastatic stage only (p < 0.03). Unstimulated PBMC manifested decrease in IL-2 (p < 0.03) and increased level of TNF-alpha in advanced disease (p < 0.02). This impairment reflected tumor size and differentiation stage. Serum concentrations of IL-2, sIL-2R and TNF-alpha were within normal range. However, in relation to the clinical stage, significantly increased serum IL-2 was noted in combined Stage I and II as compared to controls (p = 0.012). IL-6 and CRP showed markedly elevated levels with a significancy which allowed to distinguish samples from metastatic patients. In conclusion careful comparisons of these data with clinical course of cytokine treated patients will disclose which of those tests may possess predictive power in the individual patients who are likely to respond to cytokine-based treatment.

p53 protein expression in resected invasive esophageal cancer.

Many studies have reported the increased expression of p53 protein in various human malignancies and its accumulation have been considered an intermediate biomarker in multistage carcinogenesis. This study was designed to evaluate p53 expression by immunohistochemistry using Dako p53, D0-7 monoclonal antibody in 33 resected invasive squamous cell esophageal cancers (SqCC). The relationship between p53 immunoreactivity and clinicopathologic parameters was determined by the Chi-square test and Student's t test. p53 protein overexpression (more then 10% positive staining cancer cells) was found in 15 out of 33 (45%) tumors. Positive test was found in 38% cases in Stage IIA, 57% in Stage IIB, 45% in Stage III and 50% cases in Stage IV. p53 overexpression was observed in 48% of tumors with lymph nodes metastases, and 41% of tumors without lymph nodes metastases. In respect of tumor differentiation, cases graded as G1, G2 and G3 were positive in 50%, 50% and 40%, respectively. Thirteen per cent of patients with p53 protein overexpression and 16% of patients without p53 protein overexpression survived more than 3 years. There was no correlation between p53 overexpression and stage, tumor differentiation, lymph nodes metastases, and patients survival. In conclusion our results showed that p53 overexpression did not correlate with clinicopathologic feature of invasive SqCC of the esophagus and p53 protein overexpression was unsuitable for predicting the outcome of patients after surgical resection.

Potential carcinogenicity of the synthetic 1,3,6-triazine (6-azapyrimidine) nucleic acid analogues determined by DC polarography. II. Nucleosides of 6-azauracil.

The polarographic reduction of six synthetic 1,3,6-triazine (6-aza) nucleosides with 6-azauracil as the nucleoside base in the 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 compared for six nucleosides of 6-azauracil and two nucleosides of 4-thio-6-azauracil. The current value of the first diffuse polarographic wave or a new diffuse polarographic wave belonging to the nucleoside-alpha-lipoic acid complex increased with the increase of the alpha-lipoic acid concentration for the all compounds only marginally. Although this diffuse current increase was linear and dependent on the alpha-lipoic acid concentration in anhydrous solutions, the determined index tg alpha values ranged between 0.027 and 0.114. This is an indication of a very low potential carcinogenicity of the all nucleoside analogues investigated.

Transformation kinetics of the 5'-chloro-5'-deoxy analogue of arabinosylcytosine.

5'-Chloro-5'-deoxy arabinosylcytosine (Cl-araC) is a more lipophilic analog of the clinically used drug--arabinosylcytosine (araC). The resistance toward the enzyme cytidine-deaminase action was described as an characteristic feature of this synthetic nucleoside. The kinetics of the Cl-araC transformation in acid and alkaline solutions was studied at various temperatures. When compared with parent compound araC, the chlorine atom at the 5' position of the nucleoside sugar moiety increases the Cl-araC stability. The chlorine atom stabilizing effect is higher in acidic conditions. Cl-araC increased stability, antileukemic activity accompanied by higher lipophilicity confirm the fact that Cl-araC belong among interesting compounds from the point of view of cancer chemotherapy.

The increase of the rate of hemopoietic recovery and clinical benefit of the erythropoietin (EPO) and granulocyte colony-stimulating factor (G-CSF) with peripheral blood progenitor cells (PBPC) after intensive cyclic chemotherapy in high-risk breast cancer patients.

The role of erythropoietin (EPO) plus granulocyte-colony stimulating factor (G-CSF) combination in hemopoietic recovery was studied in patients with high-risk breast carcinoma and compared to a control group of previously treated identical patients who were not given EPO plus G-CSF. Eleven consecutive patients admitted to this study had Stage III or IV breast cancer. They received 6 cycles of intensive chemotherapy (epirubicin 150 mg/m2 and cyclophosphamide 1300 mg/m2). The 1st cycle served for mobilization of peripheral blood progenitor cells (PBPC). At its end leukaphereses collections of PBPC were performed to be used as hematologic support (PBPCT) in the 5 remaining cycles. The administration of EPO plus G-CSF was started when leukocyte (WBC) count in peripheral blood dropped below 1 x 10(9)/l and hemoglobin (Hb) level fell below 100 g/l. The treatment was stopped when leukocyte count rose to 5 x 10(9)/l and Hb to 130 g/l. EPO plus G-CSF combination after PBPCT produced significant effects in terms of hemopoietic recovery, clinical benefit and supportive care requirements when compared with 12 historic control patients: Periods of leukopenia were shorter which resulted in reduced risk of infectious complications. The grades of leukopenia in the study and control groups were as follows: grade 4 (36 vs. 18%), grade 3 (57 vs. 30%), grade 2 (7 vs. 13%) respectively. Significantly shorter was the time of PLT recovery < 50 x 10(9)/l (p < 0.001). The grades of thrombocytopenia were: grade 4 (29 vs. 11%), grade 3 (21 vs. 12%), grade 2 (25 vs. 36%) respectively. The number of necessary transfusions was significantly reduced as well as the length of hospital stay (p < 0.001). In conclusion, our results obtained in this study confirm that combination of EPO plus G-CSF not only increases the rate of hemopoietic recovery, reduces the number of necessary red blood cell and platelet transfusions but, at the same time, simplifies the clinical management and is more tolerable for the patients.

Immunohistochemical analysis of HLA class II antigens and tumor infiltrating mononuclear cells in renal cell carcinoma: correlation with clinical and histopathological data.

Cryostat sections of 37 renal cell carcinomas (RCC)--25 clear cell type, 10 granular and 2 chromophobe--were studied with indirect immunoperoxidase method applying monoclonal antibodies (MoAb) to HLA-DR, -DP and -DQ antigens for analysis of HLA class II antigens, and anti-CD14, -CD3, -CD4 and -CD8 MoAb for tumor infiltrating mononuclear cells (TIM). Number of positive cells was estimated semiquantitatively and results of immunohistochemical investigation were correlated with clinical (patient age and sex, tumor size and TNM stage) and histopathological (cytology, histology, grade) characteristics of RCC. All RCC expressed HLA-DR, 92% -DQ and 73% -DP antigens with level of expression in hierarchy- DR>-DQ>-DP, but no statistically important correlation could be established with any of the histopathological or clinical parameters analyzed. Monocytes were more abundant than T lymphocytes and CD4+ than CD8+ T cells, whereas tumors with T lymphocyte predominance and approximately equal number of CD4+ and CD8+ T cells had greatest average diameter. Inadequate activation of T lymphocytes by tumor cells (despite capability of antigen presentation) could be the reason for association of parameters which indicates more aggressive tumor behavior with aberrant HLA class II antigen expression on RCC.

Changes after conservative surgical treatment for early breast carcinoma: comparison of sonographic and bioptic findings.

The aim of the study was evaluation of interventional ultrasound in the diagnostics of changes after conservative surgical treatment for early breast carcinoma and in the therapeutic removal of postoperative seromas. The analysis of USG- and bioptic findings in 240 breast cancer patients after conservative breast surgery performed between 1996-1998 is reported. The major findings were: postoperative scars, granulomas, seromas, hematomas, fat necrosis, skin thickening, inflammatory changes and local tumor recurrence. In our experience, the interventional ultrasound coupled with cytological examination are reliable methods for the follow up of early and late postoperative changes with sufficient degree of correlation between USG- and histologic findings.

Sole brachytherapy of the tumor bed after breast conserving surgery: a new radiotherapeutic strategy for patients at low risk of local relapse.

The aim of the study was to test the hypothesis, if there were subgroups of early breast cancer patients in which sole brachytherapy (BT) of the tumor bed was a feasible and safe treatment option after breast conserving surgery (BCS). Forty four prospectively selected patients with Stage I-II breast cancer were entered into a protocol of postoperative tumor bed irradiation using interstitial high dose rate (HDR) implants. The HDR fractionation schedules were calculated according to the linear quadratic model. In 8 patients 7 x 4.33 Gy, in the other 36 patients 7 x 5.2 Gy were delivered to the tumor bed with 2 cm margin. The treatment planning was based on the 3 dimensional (3D) reconstruction of the clipped excision cavity, catheters and skin points. A conformal semi-3D dose planning was used. The side effects were assessed by mammograms, MRI- and clinical examinations. At a median follow up of 20 (7-36) months 1 (2.3%) local and 1 (2.3%) regional failure was observed. Distant metastasis did not occur. The cosmetic results were judged to be excellent in each case. G2 radiation side effects were observed in 2 (4.5%) cases. Postoperative sole BT of the tumor bed with careful patient selection and adequate quality assurance seems to be a feasible alternative to whole breast radiotherapy after BCS. Sole BT shortens the time of radiotherapy from 5-6 weeks to 5 days, and reduces the costs of treatment. The skin and volume sparing effect of interstitial irradiation may decrease the side effects of radiotherapy. A randomized study is in progress to define which subgroups of patients should be candidates for BT alone after BCS.

Drug dose delivery and treatment outcome relationship in standard bleomycin, etoposide and cisplatin combination chemotherapy in nonseminomatous germ cell tumor patients.

This study retrospectively evaluated the influence of drug dose delivery components (DDDC) of bleomycin, etoposide and cisplatin chemotherapy for metastatic nonseminomatous germ cell tumors on treatment outcome (NSGCT). Between December 1987 and January 1995, 75 NSGCT patients were treated with a median of 4 cycles (range 3-8) of cisplatin 120 mg/m2 on day 1, etoposide 100 mg/m2 on days 1 through 5 and bleomycin 30 U on days 1, 3, and 5 every 3 weeks. DDDC, such as cumulative dose, cumulative dose in mg/m2, dose intensity (DI), relative dose intensity (RDI), dose intensity products, and relative dose intensity products by drug, were calculated and tested as possible predictors of treatment outcome in patients classified according to Indiana University (IU), and International Germ Cell Cancer Cooperative Group (IGCCCG) classifications. Overall complete response (CR) rate was 64%, and 3-year progression-free survival (PFS) was 59%. By IU classification there were statistical differences in CR and survival between moderate (89-81%) and advanced disease (42-40%) (p < 0.005), while for patients classified according to IGCCCG criteria, statistical differences in CR and PFS there were not registered. DI (mg/m2/week) and RDI values for the entire group were: cisplatin 33-0.82; etoposide 133-0.80 and bleomycin 11-0.37. We did not observe a statistically significant difference in drug dose delivery components for treatment outcome between patients who achieved a CR and incomplete response when analyzed by either extent of disease or whole group. Extent of disease was the most important predictor of treatment outcome.

Enhancement of radiosensitivity in human glioblastoma U138MG cells by tetrandrine.

Tetrandrine, a natural lipid-soluble alkaloid with a low molecular weight, has both anti-tumor activity and a tissue-protective effect, as demonstrated in our previous studies. We studied tetrandrine to determine whether or not it enhanced radiosensitivity in human glioblastoma U138MG cells. Tetrandrine at concentration of 5.0 and 7.5 eg/ml dramatically enhanced the growth-inhibiting effect of radiation. This effect is dose-dependent on the dose of both tetrandrine and radiation. Moreover, we found that tetrandrine eliminated the cell cycle perturbation induced by radiation. Cells treated with tetrandrine alone as well as with a combination of tetrandrine and radiation became rounded, floated up and grew sparsely under observation under an inverted light microscope, DNA fragmentation was also noted by gel electrophoresis. These findings suggest that tetrandrine has potential as an adjunct to radiotherapy for glioblastoma.

Diazenes: modificators of tumor cell resistance to cisplatin.

Most studies indicate that modulation of glutathione metabolism may be one of the most promising means of reversing clinical drug resistance. Five new diazene compounds have been synthesized: JK-279, JK-835, JK-913, JK-925 and LV-57 that should, according to their structure and biochemical properties, lower the GSH concentration. In the present study, we examined the influence of diazenes on cisplatin resistance in human cervical (HeLa) and laryngeal carcinoma (HEp2) cells as well as in their cisplatin-resistant sublines (HeLaCA and CK2, respectively). Intracellular GSH content was examined spectrophotometrically by the procedure developed by Tietze. The cell sensitivity to drugs was determined using a modified colorimetric MTT assay. Results show that all examined diazenes lowered GSH concentration. This decrease was insignificant for JK-835 and JK-925 in HeLa and HeLaCA cells, and JK-925 in CK2 cells. In human cervical carcinoma HeLa and HeLaCA cells, JK-279 was mostly active in sensitizing the cells to cisplatin, especially in drug-resistant cells. JK-913, JK-835 and LV-57 reverted partially resistance to cisplatin in HEp2 cells, while none of the diazenes was active in CK2 cells. In conclusion, diazene JK-279 may be useful in the combined treatment (cisplatin + diazene) for the certain type of cancer.

Achiral and chiral HPLC analysis of norverapamil and its metabolites in microsomal samples.

Achiral and chiral HPLC (high performance liquid chromatography) were developed for monitoring norverapamil and its two metabolites in in vitro microsomal samples. Norverapamil is a chiral drug with the same ability as verapamil in reversing MDR (multi-drug resistance) against some cytostatics. R(+) norverapamil is less cardiotoxic as S(-) enantiomer and so it could be applied in higher concentrations. HPLC conditions were worked out for both achiral and chiral assays and very simple and effective preseparation procedure (SPE - solid phase extraction) was used for microsomal sample as clean-up step before HPLC analysis. The total concentrations of norverapamil and its metabolites were calculated from achiral chromatograms and the ratio of enantiomers were determined from chiral separations. The complete assay could be used for both in vitro and in vivo experiments.