Adriamycin in combination with dexamethasone and octreotide lacks activity on the treatment of a 4T1 metastatic breast cancer model.

The aim of this study was to evaluate whether the palliative treatment for metastatic disease with dexamethasone (DEX) plus octreotide (OCT) can improve the anticancer effects of the standard treatment with adriamycin (ADR) on a 4T1 metastatic breast cancer (MBC) model. 4T1 cells were first characterized for the expression of the somatostatin receptors 1-5 and were then inoculated onto the femur of BALB/C mice. Investigation protocols used 4T1 cell proliferation and invasion assays, analysis of radiographic images of the bone metastatic lesions, and overall survival of the diseased animals. The triple combination treatment regime (ADR+OCT+DEX) was ineffective for growth inhibition and showed an antagonistic effect on ADR activity in the 4T1 cell line in both proliferation and invasion assays. ADR treatment following the administration of the DEX+OCT regimen decreased the anticancer activity of ADR both on the grading of the bone metastatic lesions and on the overall survival of diseased animals. Moreover, the palliation treatment with OCT+DEX and in combination with ADR rather caused disease progression of the metastatic disease and bone lesions in a 4T1 MBC model in vivo. These results suggest that the administration of the DEX+OCT regimen, although may preserve palliative effects, neutralizes or reverses the anticancer effects of ADR on a 4T1 MBC model in vitro and in vivo. The simultaneous use of these drugs should be considered carefully in clinical practice.

Plasma indoleamine 2,3-dioxygenase concentration is increased in hemodialysis patients and may contribute to the pathogenesis of coronary heart disease.

INTRODUCTION: Coronary heart disease (CHD) is the leading cause of death in hemodialysis (HD) patients. Inflammation contributes to the pathogenesis of atherosclerosis in this population. Indoleamine 2,3-dioxygenase (IDO), an enzyme with immunomodulatory properties, was evaluated in HD patients with or without CHD. METHODS: Of the total of 66 HD patients, 22 of them with CHD were confirmed by coronary angiography and 24 healthy volunteers were enrolled in the study. Plasma IDO was assessed by means of enzyme-linked immunosorbent assay. Serum interleukin-6 (IL-6) and C-reactive protein (CRP) were also measured. RESULTS: Compared with healthy volunteers, plasma IDO concentration was markedly increased in HD patients (median 8.04 ng/mL vs. 48.9 ng/mL). Serum IL-6 and CRP were also significantly increased in HD patients. Compared with HD patients without CHD, plasma IDO concentration was significantly increased in HD patients with CHD (median 38.6 ng/mL vs. 74.5 ng/mL). Neither IL-6 nor CRP differed between the last two groups. IDO was negatively correlated with IL-6 and CRP. CONCLUSION: IDO concentration is increased in HD patients and is increased further in HD patients with CHD. It remains to be elucidated if increased IDO plays a direct role in the pathogenesis of atherosclerosis or if it affects atherosclerosis indirectly by curtailing chronic inflammation or both.

Plasma indoleamine 2,3-dioxygenase and arginase type I may contribute to decreased blood T-cell count in hemodialysis patients.

BACKGROUND: Acquired immunity is impaired in hemodialysis (HD) patients, and decreased T-cell number may contribute. Indoleamine 2,3-dioxygenase (IDO) and arginase type I (ARG) catabolize tryptophane and arginine, respectively, and exert proapoptotic and antiproliferative effects on T-cells. Plasma levels of IDO and ARG and their relation to blood T-cell number were evaluated in HD patients. METHODS: Thirty-two HD patients and 20 healthy controls participated in the study. Plasma IDO and ARG were measured by means of enzyme-linked immunosorbent assay. T-cell number was assessed by means of flow cytometry. RESULTS: IDO concentration was significantly higher in HD patients than in healthy volunteers (44.30 ± 31.83 ng/mL vs. 21.28 ± 26.21 ng/mL, p = 0.009). There was a trend for higher ARG concentration in HD patients (13.43 ± 11.91 ng/mL) than in healthy volunteers (9.56 ± 4.03 ng/mL), which, however, did not reach statistic significance (p = 0.099). Absolute T-cell count was significantly lower in HD patients than in healthy controls (1176.99 ± 567.71 cells/mm3 vs. 1519.85 ± 594.96 cells/mm3, p = 0.040). Absolute blood T-cell number was inversely related to plasma IDO (r = -0.490, p = 0.004) and to plasma ARG (r = -0.387, p = 0.029) concentrations. CONCLUSIONS: Plasma IDO and ARG may contribute to decreased blood T-cell count in HD patients.

Vitamin D receptor activators and response to injury in kidney diseases.

Clinical studies have confirmed that administration of vitamin D receptor (VDR) activators offers a survival benefit in hemodialysis patients and may help in preservation of renal function in predialysis patients. Accumulated clinical and mainly experimental data support that in the context of kidney disease, VDR activators exert their beneficial effect not only due to their action on calcium and phosphorus homeostasis, but also through modulation of the response to injury. They attenuate systemic and renal inflammation, and they affect the tissue repair process, reducing renal fibrosis. This aspect of the functions of VDR activators in kidney disease is reviewed in the present manuscript.

Thyroid carcinoma in children and adolescents: presentation, clinical course, and outcome of therapy in 23 children and adolescents in Northern Greece.

OBJECTIVE: To define clinical presentation, surgical complications, follow-up characteristics, and survival of 23 children and adolescents with thyroid carcinoma. DESIGN: Retrospective analysis of data of 23 children and adolescents with thyroid carcinoma cared for in the Theagenion Cancer Hospital, Thessaloniki. RESULTS: At the time of diagnosis cervical lymph node metastases were present in 18 (78.2%), mediastinal in 2 (8.69%), and pulmonary in 3 patients (13%). Total thyroidectomy was performed in 21 patients, with lymph node dissection in 18 and subtotal thyroidectomy in 2. No significant post-operative complications were observed. Histological examination revealed differentiated thyroid carcinoma (DTC). Tumor was multifocal in 11 (47.8%) and bilateral in 7 subjects (63.6%). Thyroid capsule invasion, vascular invasion, soft tissue involvement, and parathyroid gland invasion was observed in 12 out of 23 patients (52.1%). All patients received thyroxine suppressive therapy and 21 of them additional therapy with radioactive iodine (131I). During follow-up (5.5 years), 6 out of the 23 patients presented new metastases in the cervical and mediastinal lymph nodes, lungs, and bones. At last follow-up, 11 patients (47.8%) had residual disease but all were alive. CONCLUSIONS: Thyroid carcinoma in children and adolescents can be quite aggressive. When a young patient presents thyroid nodule or multinodular goiter or palpable lymph nodes, the existence of thyroid carcinoma must be seriously considered.

Increased plasma angiogenin level is associated and may contribute to decreased T-cell zeta-chain expression in hemodialysis patients.

Hemodialysis (HD) patients are characterized by impaired T-cell function at least in part because of T-cell zeta-chain downregulation due to inflammation. Angiogenin responds as an acute phase reactant, is increased in HD patients, suppresses T-cell function and increased angiogenin level is co-localized with T-cell zeta-chain downregulation in various pathologies. Angiogenin can inhibit translation of proteins, which lack internal ribosomal entry sites in the corresponding mRNAs. In this study, the possible effect of angiogenin on T-cell zeta-chain downregulation was evaluated. Thirty HD patients and 21 healthy volunteers participated. T-cell zeta-chain expression was assessed with flow cytometry, plasma angiogenin and serum IL-6 with ELISA and serum C-reactive protein with an immunoturbidimetric method. Two available software tools were used for predicting the presence or not of internal ribosomal entry sites in T-cell zeta-chain mRNA sequence. In silico analysis of T-cell zeta-chain mRNA sequence failed to reveal the presence of internal ribosomal entry sites. T-cell zeta-chain expression was lower in HD patients than in healthy volunteers (1.86 ± 0.63 vs. 4.73 ± 3.22). In HD patients, C-reactive protein as well as IL-6 were higher than in healthy volunteers (10.04 ± 15.13 mg/L vs. 3.43 ± 0.98 mg/L and 15.06 ± 13.08 pg/mL vs. 2.11 ± 2.10 pg/mL respectively). Angiogenin was higher in HD patients than in healthy volunteers (483.20 ± 154.07 ng/mL vs. 259.98 ± 64.15 ng/mL). Neither C-reactive protein, nor IL-6 was associated with angiogenin or T-cell zeta-chain expression. Angiogenin concentration was negatively related to the expression of T-cell zeta-chain (r = -0.410, P = 0.025). Increased angiogenin may contribute to decreased T-cell zeta-chain expression in HD patients.

Serum osteoprotegerin is markedly increased and may contribute to decreased blood T cell count in hemodialysis patients.

PURPOSE: Acquired immunity is impaired in hemodialysis (HD) patients, and decreased T cell number may contribute. Receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) are expressed in cells of the immune system and affect T cell development, homeostasis and proliferation. Serum levels of RANKL and OPG and their relation to blood T cell number were evaluated in HD patients. METHODS: Thirty-four HD patients and 20 healthy controls participated in the study. Serum RANKL and OPG were measured by ELISA and T cell number was assessed by flow cytometry. RESULTS: Median RANKL concentration did not differ between HD patients and healthy volunteers (300.00 pmol/L; range, 3,340.00 vs. 330.00 pmol/L; range, 440.00 pmol/L; p = 0.528). Median OPG was markedly higher in HD patients than in healthy volunteers (13.12 ± 4.71 vs. 4.71 ± 0.93 pmol/L, p < 0.001). The T cell count was significantly lower in HD patients than in healthy controls (1,177.00 ± 567.71 vs. 1,519.80 ± 594.96 cells/mm(3), p = 0.044). In HD patients, blood T cell number was correlated positively with serum RANKL (ρ = 0.462, p = 0.007) and negatively to serum OPG (r = -0.449, p = 0.008). CONCLUSION: Serum OPG concentration is markedly increased in HD patients and may contribute to decreased blood T cell count and impaired acquired immunity that characterizes this population.

Plasma serotonin and markers of bone formation and bone resorption in hemodialysis patients.

INTRODUCTION: Serotonin receptors are present in osteoblasts and osteoclasts, and serotonin affects bone metabolism. The association of plasma serotonin with markers of bone formation and bone resorption in hemodialysis patients was evaluated. MATERIALS AND METHODS: Twenty-four hemodialysis patients (11 diabetics) and 22 healthy volunteers were enrolled into the study. Serotonin was assessed in platelet-free plasma, whereas the markers of osteoblastic activity N-terminal midfragment osteocalcin and total procollagen type-1 aminoterminal propeptide as well as the marker of osteoclastic activity beta-isomerized C-terminal cross-linked peptide of collagen type I were measured in serum. Serum intact parathyroid hormone was also assessed. RESULTS: Serotonin did not significantly differ between hemodialysis patients and healthy volunteers. All evaluated markers of bone metabolism and intact parathyroid hormone were much higher in hemodialysis patients. Serotonin was significantly correlated with all evaluated markers of bone metabolism in hemodialysis patients. Serotonin was reversely related to the patients' age. Serotonin, osteocalcin, procollagen type-1 aminoterminal propeptide, and beta-isomerized C-terminal cross-linked peptide of collagen type I were much lower in diabetic hemodialysis patients. CONCLUSIONS: Serotonin may increase both bone formation and bone resorption in hemodialysis patients. The reverse relation of serotonin to patients' age as well as its lower levels in diabetic hemodialysis patients indicate that low plasma serotonin may contribute to the higher incidence of low-turnover bone disease that characterizes old and diabetic hemodialysis patients.

Docetaxel-carboplatin in combination with erlotinib and/or bevacizumab in patients with non-small cell lung cancer.

BACKGROUND: Bevacizumab and erlotinib have been demonstrated to prolong overall survival in patients with non-squamous non-small cell lung cancer (NSCLC). We designed a four-arm Phase III trial to evaluate the efficacy and toxicity of the combination of docetaxel, carboplatin, bevacizumab, and erlotinib in the first-line treatment of patients with NSCLC. METHODS: A total of 229 patients with stage IIIb/IV non-squamous NSCLC were treated with two cycles of carboplatin (area under the concentration-time curve 5.5) and docetaxel 100 mg/m2 as chemotherapy. After completion of two treatment cycles, patients were evaluated for response and divided into four groups: 61/229 continued with four more cycles of chemotherapy (control group), 52/229 received chemotherapy plus erlotinib 150 mg daily, 56/229 received chemotherapy plus bevacizumab 7.5 mg/kg, and 60/229 were treated with the combination of chemotherapy, erlotinib, and bevacizumab until disease progression. The primary endpoint was overall survival. RESULTS: Over 4 years of follow-up, there was no statistically significant difference in survival and time to progression between the four treatment groups. After two cycles of chemotherapy, responders and nonresponders were divided according to their response in order to examine the role of initial response as an independent factor in survival and response when a biological agent is combined with chemotherapy. Nonresponders, who received additional therapy with bevacizumab or combination therapy, had a survival benefit [657 days (95% confidence interval 349-970) and 681 days (95% confidence interval 315-912), respectively], which was statistically significant compared with continuation of cytotoxic chemotherapy (P < 0.001). The combination therapy had a safety profile comparable with that of bevacizumab and erlotinib taken individually. CONCLUSION: Administration of bevacizumab and erlotinib in combination with first-line chemotherapy, followed by bevacizumab and erlotinib monotherapy as maintenance, showed promising results in patients with NSCLC, with reduced toxicity as compared with chemotherapy alone, but did not translate into longer overall survival.

Gonadotropin-releasing hormone neuropeptides and receptor in human breast cancer: correlation to poor prognosis parameters.

Expression of the two gonadotropin-releasing hormone homologue peptides GnRHI and GnRHII and their receptor GnRHR has been demonstrated in a number of malignancies. In hormone-dependent breast cancer, GnRH analogs are used for therapy in premenopausal women. Gene expression of GnRHI, II and R was studied in breast biopsies from primary breast adenocarcinoma obtained from the tumor and the adjacent benign tissue. Levels were evaluated by a multiplex real-time RT-PCR. GnRHI transcripts were detected in 14.7% of the benign and 29.4% malignant biopsies and GnRHII in 21.2% benign and 44.1% malignant biopsies. GnRHR was also more frequent in the malignant (54.2%) than in the benign (24.0%) biopsies, at similar expression levels. No transcripts were detected in biopsies from healthy individuals. There was a strong correlation between the presence of GnRHI and GnRHII transcripts and their receptor in the benign and the malignant biopsies. GnRHI, II and R expression correlated significantly with poor prognosis pathological parameters. Immunohistochemistry for GnRHR revealed expression in malignant cells and in epithelial cells of mammary ducts of the adjacent area with pre-cancerous features. In contrast, GnRH I and II peptides were rarely expressed at low levels in breast cancer cells. In conclusion GnRH peptides and receptor are expressed more frequently in breast tumors than in the adjacent mammary tissue, representing a malignant feature. Their expression correlated to tumor characteristics of poor prognosis and was therefore related to more aggressive malignancies. Concomitant expression of peptides and receptor supports an autocrine/paracrine regulating role.

Inverse association of serum 25-hydroxyvitamin D with markers of inflammation and suppression of osteoclastic activity in hemodialysis patients.

INTRODUCTION: In hemodialysis patients, 25-hydroxyvitamin D conversion to active 1,25-dihydroxyvitamin D by the kidneys is very limited. The expression of both vitamin D receptor and 1alpha-hydroxylase in cells of the immune system and in both osteoblasts and osteoclasts makes it possible that 25-hydroxyvitamin D could play an important role in both inflammation and bone metabolism acting in a autocrine and/or paracrine way in these patients. MATERIALS AND METHODS: Thirty-three hemodialysis patients not under vitamin D receptor agonist treatment were enrolled into the study. Serum levels of 25-hydroxyvitamin D, C-reactive protein (CRP), interleukin-6 (IL-6), receptor activator of nuclear factor-kappaB ligand (RANKL), and osteoprotegerin, as well as intact parathyroid hormone were assessed by immunoassays. RESULTS: Regarding inflammation, 25-hydroxyvitamin D inversely correlated with both CRP and IL-6. Regarding bone metabolism, 25-hydroxyvitamin D was positively related to osteoprotegerin, but negatively to the RANKL. The latter could be the result of parathyroid hormone suppression by 25-hydroxyvitamin D, since 25-hydroxyvitamin D negatively correlated with parathyroid hormone, which in turn was positively related to RANKL. CONCLUSIONS: Serum 25-hydroxyvitamin D is inversely correlated with markers of inflammation and may suppress osteoclastic activity in hemodialysis patients.

Plasma angiogenin and vascular endothelial growth factor a among hemodialysis patients.

INTRODUCTION: Angiogenesis plays a role in the pathogenesis of coronary heart disease (CHD) and diabetes mellitus (DM) pathology, and certain angiogenic factors are increased by inflammation. The aim of this study was to evaluate plasma angiogenin and vascular endothelial factor A (VEGFA) levels in hemodialysis patients, as well as the effect of CHD, DM, and inflammation on these markers. MATERIALS AND METHODS: Sixty-six hemodialysis patients were enrolled in the study, of whom 22 (33.3%) suffered from CHD , 22 (33.3%) from DM, and 28 (42.4%) from inflammation. They were compared with 24 healthy volunteers. Plasma angiogenin and VEGFA were assessed by means of enzyme-linked immunosorbent assay, and serum C-reactive protein was measured with an immunoturbidimetric method. These markers were compared between hemodialysis patients with and without CHD, DM, and inflammation. RESULTS: Compared to healthy volunteers, plasma angiogenin was significantly higher in hemodialysis patients (263.57 ± 65.95 ng/mL versus 499.15 ± 175.68 ng/mL; P < .001). Similarly, plasma VEGFA was markedly increased in hemodialysis patients (median, 60.50 pg/mL; range, 280 pg/mL), compared to healthy volunteers (median, 28.84 pg/mL; range, 59.40 pg/mL; P < .001). Neither angiogenin nor VEGFA levels differed significantly between hemodialysis patients with and without CHD, DM, or inflammation. CONCLUSIONS: Plasma angiogenin and VEGFA levels are markedly increased in hemodialysis patients, but not associated with CHD, DM, or inflammation among hemodialysis patients.

Perilipin-1 in hemodialyzed patients: association with history of coronary heart disease and lipid profile.

Perilipin-1 surrounds lipid droplets in both adipocytes and in atheroma plaque foam cells and controls access of lipases to the lipid core. In hemodialysis (HD) patients, dyslipidemia, malnutrition, inflammation and atherosclerosis are common. Thirty-six HD patients and 28 healthy volunteers were enrolled into the study. Ten HD patients suffered from coronary heart disease (CHD). Perilipin-1, triglycerides, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol (HDL-C), body mass index, albumin, geriatric nutritional risk index, normalized protein catabolic rate, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured. Perilipin-1 did not differ between HD patients and healthy volunteers. IL-6 and TNF-α were higher in HD patients. The evaluated nutritional markers and the markers of inflammation did not differ between HD patients with high perilipin-1 levels and HD patients with low perilipin-1 levels. Regarding the lipid profile, only HDL-C differed between HD patients with high perilipin-1 levels and HD patients with low perilipin-1 levels, and it was higher in the first subgroup. Perilipin-1 was significantly higher in HD patients without CHD. Perilipin-1 is detectable in the serum of HD patients and it is associated with increased HDL-C and decreased incidence of CHD.

Arginase type I as a marker of coronary heart disease in hemodialysis patients.

BACKGROUND: Cardiovascular disease, and mainly coronary heart disease (CHD), is the leading cause of death in hemodialysis (HD) patients. Non-traditional risk factors may play an important role in this population. Arginase is known to contribute directly to atherosclerosis progression and to counteract the beneficial effects of nitric oxide. HD could be considered as an inflammatory condition. Inflammation contributes to atherosclerosis progression and influences both arginase and nitric oxide synthase expression. In the present study, serum arginase type I was evaluated as a marker of CHD in HD patients. The markers of inflammation interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were also assessed. PATIENTS AND METHODS: Sixty-eight HD patients and 24 healthy volunteers were enrolled into the study. Twenty HD patients suffered from CHD confirmed with coronary angiography, while the remaining 48 HD patients were asymptomatic. Serum arginase type I, IL-6 and TNF-α were measured with ELISA. During 24 months follow-up, none of the asymptomatic subjects developed symptoms of CHD. RESULTS: IL-6 and TNF-α levels were increased in HD patients, but did not differ between HD patients with or without CHD. On the contrary, arginase levels did not differ between healthy subjects and HD patients, but were twice higher in HD patients with CHD than in HD patients without CHD (22.41 ± 15.47 ng/ml vs. 10.16 ± 8.13 ng/ml). CONCLUSION: Arginase type I may contribute to the pathogenesis of CHD in HD patients and its serum levels could be used as a marker of CHD in this population.

Indoleamine 2,3-dioxygenase is increased in hemodialysis patients and affects immune response to hepatitis B vaccination.

BACKGROUND: Acquired immunity is impaired in hemodialysis (HD) patients. Indoleamine 2,3-dioxygenase (IDO) is inducible by inflammation and through tryptophan depletion and generation of kynurenine pathway products suppresses adaptive immune response. In the present study plasma IDO levels were assessed in HD patients. Its effect on response to HBV vaccination program was evaluated. PATIENTS AND METHODS: Sixty-six HD patients and twenty-four healthy volunteers enrolled into the study. All the HD patients were initially vaccinated with four double doses of recombinant HBV vaccine. All doses were repeated in patients who had not responded after complete first vaccination series. Only one boost dose was being administered in patients with initial adequate antibody levels against the HBV surface antigen (anti-HBs>10I U/L) who then presented with reduced anti-HBs levels. IDO, CRP, IL-6 and TNF-α were measured by means of ELISA. RESULTS: Compared to healthy volunteers, IDO levels were twice higher in HD patients. CRP, IL-6 and TNF-α were also much higher in HD patients. IDO levels were almost twice higher in HD patients with inadequate response to HBV vaccination, than in those with adequate immune response. CRP, IL-6 and TNF-α did not differ between the two patients' groups. IDO was negatively correlated with all markers of inflammation in HD patients. CONCLUSION: IDO is increased in HD patients. It is possible that after its initial upregulation due to chronic inflammation, IDO curtails its own provoking agent, i.e., inflammation. Increased IDO suppresses adaptive immunity in HD patients, as it is assessed by the response to HBV vaccination.

Paricalcitol reduces basal and lipopolysaccharide-induced (LPS) TNF-alpha and IL-8 production by human peripheral blood mononuclear cells.

INTRODUCTION: Vitamin D and its analogues proved to exert immunomodulatory effects. Paricalcitol is a vitamin D analogue that is safe. It has been used for years in the treatment of secondary hyperparathyroidism in hemodialysis patients and, importantly, it is less calcemic than vitamin D. In this study the immunomodulatory/anti-inflammatory properties of paricalcitol were evaluated in vitro. SUBJECT AND METHODS: Ten healthy volunteers enrolled into the study. Peripheral blood mononuclear cells (PBMC) at a concentration of 10(6) cells per well were cultured for 48 h in the presence or not of lipopolysaccharide (LPS) (100 ng/ml) and in the presence or not of paricalcitol (10(-8) M). TNF-alpha and IL-8 were measured in the supernatants by ELISA. RESULTS: Basal TNF-alpha concentration (50.3 +/- 22 pg/ml) was reduced by paricalcitol (44.1 +/- 23.2 pg/ml). LPS increased TNF-alpha concentration (150.0 +/- 81.7 pg/ml), but paricalcitol reduced it (121.1 +/- 69.0 pg/ml). The effect of paricalcitol on IL-8 production was more profound. Basal IL-8 concentration (1926 +/- 455 pg/ml) was reduced by paricalcitol (1273 +/- 472 pg/ml). LPS increased IL-8 concentration (2361 +/- 385 pg/ml), but paricalcitol returned it to its basal level (1849 +/- 417 pg/ml). CONCLUSION: The in vitro inhibition of transforming growth factor alpha and interleukin 8 by paricalcitol confirms the immunomodulatory properties of this vitamin D analogue.

Patterns of cell death and cell cycle profiles of cultured WEHI 13 var fibroblasts exposed to eluates of composite resins used for direct and indirect restorations.

Previous studies have shown that in vitro exposure to single compounds released from composite resins may induce cell death. In the present study the effects of eluates from commercially available composite resins used for direct or indirect restorations were evaluated on the cell cycle progression and type of cell death of cultured WEHI 13 var fibroblasts. Cells exposed to eluates of the materials were assessed for cytotoxicity by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for cell death, for cell cycle profiles by flow cytometry, for caspase-3 biochemically and by immunocytochemistry, and for morphological changes by fluorescence microscopy with acridine orange. The direct composite resin eluates induced extensive apoptosis, followed by secondary necrosis. This was accompanied by cell enlargement, micromultinucleation, chromatin disintegration, cell cycle arrest at different phases, and caspase-3 activation. The composites for indirect restorations were much less cytotoxic at all biological end-points investigated. The findings suggest that composite resins used for direct and indirect dental restorations differ in their cytotoxic potential and their ability to affect basic cellular functions. This underlines the impact of improved polymerization with respect to their biologic behavior.

Serum CYFRA 21-1 in patients with invasive bladder cancer and its relevance as a tumor marker during chemotherapy.

PURPOSE: Previous studies have shown that serum levels of the degradation products of cytokeratins could be used as surrogate markers in the diagnosis and followup of patients with solid tumors, including tumors of the bladder. MATERIALS AND METHODS: The soluble cytokeratin 19 fragment CYFRA 21-1 was measured by solid phase radioimmunoassay in the serum of 142 patients with invasive transitional cell cancer of the bladder. Of the patients 56 had clinical stage I to III locally confined disease (T1-4aN0M0) and 86 had stage IV metastatic disease with lymph node and/or distant metastases. A control group consisted of 33 healthy volunteers. In a subgroup of 49 patients with metastatic disease receiving combined platinum based chemotherapy serum CYFRA 21-1 was determined prior to the initiation of therapy and after the documentation of response. RESULTS: Abnormal CYFRA 21-1 was observed in 7% of patients with locally invasive disease and in 66% of those with metastatic disease (p < 0.0001). There was no correlation of CYFRA 21-1 with tumor differentiation. Patients with abnormal CYFRA 21-1 showed statistically significant worse median overall survival. Moreover, in the subgroup of patients with metastatic disease receiving chemotherapy CYFRA 21-1 levels correlated with the response to treatment. CONCLUSIONS: Patients with transitional cell cancer of the bladder with evidence of distant metastases showed a significant increase in serum CYFRA 21-1. During chemotherapy CYFRA 21-1 appears to be a potentially sensitive and useful indicator for monitoring treatment response.