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Biomedical subjects

Efstathios Stathopoulos

Publications and source records attributed to Efstathios Stathopoulos.

13 recordsLinked to original sources

HER-2 DNA quantification of paraffin-embedded breast carcinomas with LightCycler real-time PCR in comparison to immunohistochemistry and chromogenic in situ hybridization.

OBJECTIVES: To compare the detection of HER-2 status by real-time PCR, on paraffin-embedded breast carcinomas, in respect to immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH). DESIGN AND METHODS: Paraffin-embedded breast carcinomas collected from 85 patients diagnosed with early stage breast cancer were analyzed for HER-2 gene amplification by real-time PCR and CISH, as well as for HER-2 protein expression by IHC. RESULTS: HER-2 gene amplification was observed in 19 (22.4%) of 85 breast cancer patients by real-time PCR and in 19 (22.4%) of 85 patients by CISH. Strong (3+) HER-2 protein over-expression was observed in 13 (15.3%) out of 85 patients. Moreover, there were 4 out of 85 (4.7%) patients that had moderate (2+) HER-2 protein over-expression, while 68 out of 85 (80%) patients had no HER-2 protein over-expression by IHC. There were strong concordance rates between real-time PCR and IHC (79/85, 92.9%, p<0.0001) and real-time PCR and CISH (77/85, 90.6%, p<0.0001). The concordance rate between the three methods was 90.6% (p<0.0001). CONCLUSIONS: Our data show that the results obtained for amplification of HER-2 by real-time PCR on the LightCycler are comparable to those obtained by IHC and CISH.

Breast Neoplasms↗

A novel combination of multiple primary carcinomas: urinary bladder transitional cell carcinoma, prostate adenocarcinoma and small cell lung carcinoma--report of a case and review of the literature.

BACKGROUND: The incidence of multiple primary malignant neoplasms increases with age and they are encountered more frequently nowadays than before, the phenomenon is still considered to be rare. CASE PRESENTATION: We report a case of a man in whom urinary bladder transitional cell carcinoma, metachronous prostate adenocarcinoma and small cell lung carcinoma were diagnosed within an eighteen-month period. The only known predisposing factor was that he was heavy smoker (90-100 packets per year). The literature on the phenomenon of multiple primary malignancies in a single patient is reviewed and the data is summarized. CONCLUSION: It is important for the clinicians to keep in mind the possibility of a metachronous (successive) or a synchronous (simultaneous) malignancy in a cancer patient. It is worthy mentioning this case because clustering of three primary malignancies (synchronous and metachronous) is of rare occurrence in a single patient, and, to our knowledge, this is the first report this combination of three carcinomas appearing in the same patient.

Journal Article↗

A morphometric approach for the evaluation of angiogenesis in acute lymphoblastic leukemia of childhood.

Angiogenesis was estimated in childhood acute lymphoblastic leukemia (ALL) by the use of a novel morphometric method. This was compared with the widely used one of microvessel density (MVD). Bone marrow biopsies were evaluated by immunostaining with anti-Factor VIII related antigen (FVIIIRAg). Angiogenesis was calculated as volume corrected microvessel density index (VC-MVDI), taking into account the bone marrow cellularity. According to our results both MVD and VC-MVDI were increased at diagnosis of ALL in comparison with the control group. However the VC-MVDI increment was not statistically significant. Therefore, VC-MVDI could be more representative of the true increase of angiogenesis, correlating better with the outcome of the disease.

Analysis of Variance↗

Correlation of breast cancer risk factors with HER-2/neu protein overexpression according to menopausal and estrogen receptor status.

BACKGROUND: Several researchers have claimed that classification of tumours on the basis of HER-2/neu overexpression or amplification may define a subset of breast cancer in which the net effect of a risk factor could be rather more obvious and its impact on breast cancer development more clear. We decided to investigate, in a group of patients from a geographical area with a low incidence of breast cancer, whether HER-2/neu positive tumours are correlated with established or suspected risk factors for breast cancer and thus to identify distinct subgroups of high risk women. METHODS: This study analysed data from patients who attended the Breast Unit at the University Hospital of Heraklion, Crete, Greece between 1996 and 2002. 384 women with primary invasive breast cancer were compared with 566 screened women who were referred to the Unit and had not developed breast neoplasm by the time the data were analysed. Risk factor data were obtained from each subject by personal interviews using a structured questionnaire. The detection and scoring of the HER-2/neu protein, estrogen and progesterone receptor expression were performed using immunochemistry. Odds ratios and 95% confidence intervals were determined by chi-square test and logistic regression analysis. Case-case odds ratios were calculated in order to measure the risk heterogeneity between HER-2/neu+ and HER-2/neu-tumours. Separate analyses were performed for premenopausal and postmenopausal women and according to estrogen receptor status. RESULTS: In multivariate analysis without HER-2/neu stratification, an increased breast cancer risk was associated with only four of the factors examined: use of oral contraceptives (OR = 4.40, 95%C.I: 1.46-13.28), use of HRT (OR = 7.34, 95%C.I: 2.03-26.53), an age at first full pregnancy more than 23 years (OR = 1.91, 95%C.I: 1.29-2.83) and body mass index more than 29 kg/m2 (OR = 3.13, 95%C.I: 2.02-4.84). Additionally, a history of abortion or miscarriage (OR = 0.56, 95%C.I: 0.38-0.82) was correlated with a decreased risk of breast cancer. In the case to case comparison only BMI >29 kg/m2 revealed a relative connection that was stronger with positive than with negative HER-2/neu tumours (ratio of OR's = 2.23, 95%C.I: 1.20-4.15, p = 0.011). This may indicate evidence of heterogeneity of a rather significant degree for this factor. In the ER negative group an age at first full pregnancy >23 years and a BMI >29 kg/m2 were associated with an increased risk in both HER-2/neu groups, but the association was significantly stronger for the latter factor in the positive HER-2/neu tumours (ratio of OR's = 2.46, 95%CI: 0.97-6.21). CONCLUSIONS: Our study did not confirm that the established or putative hormonal breast cancer risk factors differ regarding their relations with HER-2/neu+ versus HER-2/neu-breast tumours, with the exception of increased BMI. Further innovative studies with larger sample sizes are needed to examine how the status of these potentially modifiable breast cancer risk factors interacts with biological markers such as HER-2/neu oncoprotein.

Journal Article↗

Trastuzumab administration can effectively target chemotherapy-resistant cytokeratin-19 messenger RNA-positive tumor cells in the peripheral blood and bone marrow of patients with breast cancer.

PURPOSE: The detection of disseminated occult breast cancer cells in peripheral blood and bone marrow is associated with poor prognosis. Since a high proportion of these cells express the HER-2 receptor, we evaluated the effectiveness of the anti-HER-2 antibody trastuzumab (Herceptin) administration to eliminate them. EXPERIMENTAL DESIGN: Thirty patients with prior chemotherapy exposure were recruited to the study on the basis of having detectable cytokeratin-19 (CK-19) mRNA transcripts by nested reverse transcription (RT)-PCR in the peripheral blood and/or bone marrow. There were 13 patients with stage I, II, or III breast cancer and 17 with stage IV disease. They were treated in two cohorts with either 4 to 8 weekly infusions of trastuzumab at 2 mg/kg (4 mg/kg loading dose; 20 patients) or 2 to 3 infusions every 3 weeks at 6 mg/kg (8 mg/kg loading dose; 10 patients). All of the patients' samples were also analyzed for HER-2 by nested RT-PCR, but detectable HER-2 messenger RNA (mRNA) was not required for inclusion in the study. After trastuzumab infusions, patients were closely monitored by nested RT-PCR and real-time RT-PCR for the detection of CK-19 mRNA-positive cells. RESULTS: Before trastuzumab infusions, CK-19 mRNA-positive cells were detected in the peripheral blood (n = 10), bone marrow (n = 14), or both (n = 6). In 25 of 30 patients (83%), HER-2 mRNA expression was detected by nested RT-PCR in the pretrastuzumab CK-19-positive sample. After trastuzumab infusions, overall, 28 of 30 (93%) patients became CK-19 mRNA negative by nested RT-PCR and 20 of 30 (67%) by real-time RT-PCR. After a median follow-up of 6 months (range 2 to 22+), the median duration of CK-19 mRNA negativity by nested RT-PCR was 9, 12, and 6 months for stage I/II, III, and IV disease, respectively. CONCLUSIONS: Therapy-resistant CK-19 mRNA-positive cells in the peripheral blood and bone marrow can be effectively targeted by trastuzumab administration. Further studies are needed to evaluate the prognostic significance of the disappearance of these cells.

Adult↗

Primary idiopathic segmental infarction of the greater omentum: two cases of acute abdomen in childhood.

Idiopathic segmental infarction of the greater omentum (ISIGO) is a rare cause of acute abdominal pain in childhood. The authors present the case histories of 2 children treated in their department. The children underwent surgery with preoperative diagnosis of acute appendicitis with atypical clinical presentation. The definitive diagnosis of segmental infarction of the great omentum was made intraoperatively and confirmed pathologically. Excision of the infracted omentum was curative.

Abdomen, Acute↗

Spectral characteristics of acute lymphoblastic leukemia in childhood.

Although the differentiation and classification of acute leukemia are based upon cytochemical features as well as immunologic, cytogenetic, and molecular characteristics, in many cases the morphological distinction of normal lymphocytes from lymphoblasts of acute lymphoblastic leukemia (ALL) is difficult using light microscopy. In this study the distinction between normal lymphocytes and lymphoblasts of childhood ALL is proposed using their spectral characteristics. The method has been based upon the analysis and classification of optical absorption characteristics of the bone marrow cells. Spectral microscopy system is capable of capturing a great number of narrow-band images, in the wide spectral range of the optical spectrum. The analysis showed statistically significant difference (P < 0.0001) between normal lymphocytes and lymphoblasts as far as it concerns the detection, identification and mapping of their spectral absorption characteristics. Our results suggest the potential of spectral imaging as a new method for the distinction of lymphocytes from lymphoblasts in cases that with the light microscope, the morphologic differences are not visible in the bone marrow smears at diagnosis or the follow up of the children with ALL.

Adolescent↗

Ki-67 proliferation index: correlation with prognostic parameters and outcome in multiple myeloma.

The nuclear protein Ki-67 is a proliferation index, as it is expressed only by dividing cells. In this study, we investigated the clinical significance of Ki-67 determination on bone marrow biopsies of 35 patients with newly diagnosed multiple myeloma (MM). We examined the correlation of Ki-67 with other MM proliferation-related factors: interleukin-6 (IL-6), IL-10, bone marrow infiltration by plasma cells, serum lactate dehydrogenase (LDH), and beta 2 microglobulin (b2M). Ki-67 expression was also correlated with the survival rate of the patients. The results showed that Ki-67 expression increases with increasing stage of disease according to Durie-Salmon (classification stage III vs. I and II, p < 0.001). Furthermore, infiltration, IL-6, LDH, and b2M increase significantly with advancing stage of disease (p < 0.004). All parameters studied were significantly higher in patients versus controls. Ki-67 correlated with IL-6 (r: 0.422, p < 0.01), LDH (r: 0.437, p < 0.01), and b2M (r: 0.478, p < 0.004). There was a marked difference in survival between patients with MM with Ki-67 greater than 8% and patients with Ki-67 less than 8%, in favor of the latter (p < 0.07). We conclude that Ki-67 determination during routine pathological analysis of bone marrow in newly diagnosed MM could provide useful information about the proliferative activity and prognosis of the disease.

Aged↗

Bone marrow microvascular density and angiogenic growth factors in multiple myeloma.

There is evidence that angiogenesis plays an important role in the progression of multiple myeloma (MM). Hepatocyte growth factor (HGF) and tumor necrosis factor-alpha (TNF-alpha) are cytokines that potently stimulate angiogenesis. We evaluated the microvascular density (MVD) of bone marrow biopsies (after immunostaining with anti-CD34 antibodies) and serum levels of HGF and TNF-alpha in 43 patients with newly diagnosed MM. Twenty-four of these patients reached a plateau phase after treatment and were reevaluated for MVD, HGF and TNF-alpha. MVD values and serum levels of HGF and TNF-alpha were elevated in newly diagnosed MM patients in comparison with healthy controls. Pre-treatment MVD, HGF and TNF-alpha increased with advancing stage of MM disease. In patients reaching the plateau phase, a significant reduction in MVD, HGF and TNF-alpha levels occurred. A positive correlation was noted between pre-treatment MVD and serum levels of TNF-alpha and lactic dehydrogenase but not with HGF. However, HGF strongly correlated with beta2-microglobulin (beta2M), TNF-alpha and lactate dehydrogenase (LDH). We conclude that angiogenesis in MM, as expressed by the bone marrow MVD and the serum levels of angiogenic molecules such as HGF and TNF-alpha, increases with advancing clinical stage and decreases after effective chemotherapy.

Adult↗

Comparison of C-erbB2 (HER-2/Neu) immunohistochemistry results on invasive breast carcinomas. Experience of 2 Pathology departments in Turkey and Greece.

OBJECTIVE: To compare c-erbB2 (HER-2/Neu) immunohistochemical staining results obtained in 2 pathology departments and evaluate the reproducibility of staining and assessments. STUDY DESIGN: Ninety primary invasive ductal carcinoma cases constituted the material of this study. For concordant assessment, serial sections from the same tissue blocks were prepared in both Turkey and Greece and were sent to each other. Evaluation of c-erbB2 (HER-2/Neu) staining was done independently in 2 departments by experienced pathologists. Stained slides were evaluated semiquantitatively in Turkey and both semiquantitatively and automatically in Greece. RESULTS: There was complete agreement on staining density in 58 (64.4%) cases in both centers. In 15 cases there were major discordances in staining degrees, and in 17 cases there was a major discordance. Hypothetical treatment decisions based on these results showed that 82% of patients would have been handled the same way. CONCLUSION: Comparison of immunostaining patterns performed at 2 centers provided valuable data that may be used in the development of quality assurance policies. The present study showed the usefulness of multicenter comparative studies in initiating the development of guidelines.

Breast Neoplasms↗

A novel spectral microscope system: application in quantitative pathology.

In this paper, a novel spectral microscope system is presented together with a method for the quantitative assessment of the uptake by histologic samples of stains used in pathology to label tissue features of diagnostic importance. The critical component of the microscope is a variable interference filter-based monochromator. The system is capable of performing real-time spectral imaging in a plurality of spectral bands and micro-spectroscopy in any image pixel, in the spectral range 400-1000 nm. The wavelength-tuning step is 2.4-2.6 nm, while the full-width at half maximum in each step is about 1.5% of the operating central wavelength. The developed system integrates algorithms and calibration procedures for the calculation of the stain-uptake by the tissue. The acquired spectra from both stained tissue and calibration stain solutions enable the calculation of the concentration maps of the stains, even if the latter are multiple and overlap spatially and spectrally. The system was used for the quantitative mapping of the expression of estrogen and progesterone receptors in breast cancer cells. In this particular case, model validation shows that although two stains are employed, capturing of their transmittance at more than ten wavelengths is required in order to obtain an acceptable accuracy. These findings highlight the need for the development and implementation of spectral microscopy in pathology and its potential to introduce novel more reliable diagnostic criteria.

Algorithms↗

E-cadherin expression on fine-needle aspiration biopsies in primary lung adenocarcinomas is related to tumor differentiation and invasion.

UNLABELLED: E-cadherin, a cell surface molecule that mediates cell-cell adhesion in normal epithelium, has been shown in recent studies of tissue biopsy to be related to tumor differentiation and invasiveness. The aim of the study was to explore if preoperative E-cadherin expression on fine-needle aspiration biopsy (FNAB) specimens can predict cell differentiation and mediastinal lymph node spread and therefore tailor treatment in patients with primary lung adenocarcinoma. We studied prospectively the relationship between E-cadherin expression on FNABs and the pathological features of 50 cases of primary lung adenocarcinomas, which were diagnosed cytologically and confirmed histologically post-operatively. Expression of E-cadherin was found in 36 (72%) cases. Decreased expression of E-cadherin was correlated with poor grade adenocarcinomas (low grade 33% vs. 100% in high grade, p < 0.005) and lymph node metastasis (positive 43% vs. negative 93%, p < 0.005). No relationship was found between E-cadherin expression and tumor size. CONCLUSION: E-cadherin expression of FNAB specimens can be helpful in predicting tumor cell differentiation, invasiveness and defining a subpopulation of patients with primary lung adenocarcinoma with possible poor outcome, which should be taken into consideration in the proper management of the disease.

Adenocarcinoma↗