Evangelos Kouvaras

Increased HIF-1 alpha immunostaining in psoriasis compared to psoriasiform dermatitides

Background: Expression of hypoxia inducible factor 1 (HIF‐1) alpha can be linked to inflammation through reciprocal interactions with several cytokines. This finding is in accordance with the previously noted HIF‐1 alpha overexpression in psoriatic lesional keratinocytes.

INCENP (inner centromere protein) is overexpressed in high grade non-Hodgkin B-cell lymphomas.

Inner centromere protein (INCENP) is a member of the Chromosomal Passenger Complex (CPC), which is a four member protein complex essential for proper completion of mitosis and cell division (cytokinesis). Inappropriate chromosomal segregation and cytokinesis due to deregulated expression of chromosome passenger proteins may lead to aneuploidy and cancer including lymphomas. According to our knowledge this is the first study investigating immunohistochemical expression of INCENP in lymphoma cases and cancer tissues in general. Our purpose was to characterize the expression of INCENP in cases of non-Hodgkin B-cell lymphomas, to compare the immunoreactivity between low and high grades and to evaluate the correlation between INCENP and MIB-1 labeling indices. We examined INCENP and MIB-1 immunoreactivity in paraffin sections of 55 samples of non-Hodgkin B-cell lymphomas, obtained from 55 patients, 31 men and 24 women. Thirty were of high grade and 25 were of low grade. Our results showed significantly higher nuclear immunohistochemical expression of INCENP in high grade B-cell lymphomas versus low grade ones. Also INCENP expression was significantly correlated with MIB-1 labeling index. Taken together our results point to a possible association between increased INCENP immunostaining and B-cell lymphoma aggressiveness and also stress the need for further investigating the expression of INCENP and other mitotic regulatory proteins in lymphomas and other malignant neoplasms.

Aurora B kinase in Hodgkin lymphoma: immunohistochemical pattern of expression in neoplastic Hodgkin and Reed-Sternberg cells

Aurora B is a member of the chromosomal passenger complex, which is essential for proper completion of mitosis and cell division (cytokinesis). Inappropriate chromosomal segregation and cytokinesis due to deregulated expression of chromosome passenger proteins may lead to aneuploidy and cancer including lymphomas. According to our knowledge there are extremely limited studies investigating the immunohistochemical expression of Aurora B in tumor specimens of Hodgkin lymphoma. Our purpose was to characterize the expression of Aurora B in biopsies of Hodgkin lymphomas, and to evaluate the pattern of immunoreactivity in neoplastic Hodgkin and Reed-Sternberg cells (RS cells). We examined Aurora B immunoreactivity in paraffin sections of 15 samples of Hodgkin lymphomas, obtained from 15 patients, 8 men and 7 women. Ten were of nodular sclerosis type and five were of mixed cellularity. Our results showed immunoexpression of Aurora B in mononuclear lymphoid cells as well as in bi- and multinucleated RS cells. In addition, positive neoplastic cells in mitosis were observed, whereas a subpopulation without evidence of immunoreaction was also detected in each case. Taken together our results point to a possible association between Aurora B expression and mitotic deregulation in Hodgkin lymphoma, which may provide novel targets for treatment.

Smad4 and epithelial–mesenchymal transition proteins in colorectal carcinoma: an immunohistochemical study

Epithelial–mesenchymal transition (EMT) plays an important role in cancer metastasis. During EMT, tumor cells acquire the capacity to migrate and invade the stroma. Activation of the transforming growth factor-b (TGF-b) signaling pathway is of major importance for the initiation of EMT. Smad4, an essential protein of this pathway, is known to complex with multiple transcription factors (e.g. Snail-1, Slug, Twist-1), in various types of cancer, promoting the repression or activation of target genes. The role of Smad4 in colorectal cancer (CRC) is not straightforward so far. In the present study forty eight resected CRC tumor specimens were immunohistochemically examined in order to assess the expression of Smad4 and its association with E-cadherin, Snail-1, Slug, Twist-1 protein expression and with various pathological parameters. Smad4 was found to be positively correlated with Snail-1, Slug and Twist-1 expression (p < 0.001). On the other hand it was negatively correlated with the expression of E-cadherin (p < 0.001). Furthermore, lymphatic invasion could be clearly associated with Smad4 expression, a finding complying with the metastatic ability of EMT cells. In conclusion, Smad4 could be considered as a central component of EMT transition in human colorectal cancer that combines with transcriptional factors to reduce E-cadherin and alter the expression of the epithelial phenotype.

Development and validation of a reversed-phase HPLC method for licarbazepine monitoring in serum of patients under oxcarbazepine treatment

Licarbazepine is the pharmacologically active metabolite of oxcarbazepine, a drug indicated for the treatment of partial seizures and bipolar disorders. Several HPLC methods have been developed thus far but there is lack of control for interferences from antipsychotic drugs. The aim of the present study was to develop a simple, low-cost and reliable HPLC-UV method for the determination of licarbazepine in human serum in the presence of co-administered antiepileptic, antipsychotic and commonly prescribed drugs. Sample preparation consisted of a single protein precipitation step with methanol. Separation lasted ~9 min on a reversed-phase C18 column using a mobile phase composed of 50 mm sodium-dihydrogen-phosphate-monohydrate/acetonitrile (70:30, v/v) delivered isocratically at 0.9 mL/min and 30°C. Wavelength was 210 nm and calibration curve was linear with r2 0.998 over the range 0.2-50.0 μg/mL. Coefficient of variation was <5.03% and bias <-4.92%. Recovery ranged from 99.49 to 104.52% and the limit of detection was 0.0182 μg/mL. No interferences from the matrix or from antiepileptic, antipsychotic and commonly prescribed drugs were observed. The method was applied to serum samples of patients under oxcarbazepine treatment and proved to be a useful tool for the therapeutic drug monitoring of licarbazepine during monotherapy or adjunctive treatment of seizures or affective disorders.

Myxoid liposarcoma with cartilaginous differentiation: A case study with fish analysis and review of the literature.

Cartilaginous differentiation is rarely encountered in myxoid liposarcomas. To date, a small number of such cases have been described, and molecular or cytogenetic analysis was performed only in few of them. In the present study, we describe a primary myxoid liposarcoma with cartilaginous differentiation which arised in the left thigh of a 37-year-old man. Miscroscopically, the tumor consisted of areas with typical myxoid liposarcoma morphology and areas of sharply demarcated hyaline cartilage nodules. Here, we present the results of Fluorescence In Situ Hybridization (FISH) analysis that revealed the presence of FUS and DDIT3 gene rearrangements in both the liposarcomatous and cartilaginous components of the tumor. These findings confirm the neoplastic nature of the cartilage component in this rare tumor.

Anaplastic lymphoma kinase-positive diffuse large B cell lymphoma: immunohistochemical and FISH analysis of a rare tumor with unusual clinical presentation

Anaplastic lymphoma kinase-positive diffuse large B cell lymphoma represents a distinct subtype of diffuse large B cell lymphoma with a characteristic morphology, a distinct immunophenotypic profile, and recurrent cytogenetic/molecular genetic abnormalities. We report a case of anaplastic lymphoma diagnosed in the bone marrow trephine of a 24-year-old male. Histologically, the tumor cells exhibited plasmablastic morphology with expression of CD45, CD138, EMA, MUM1, and kappa immunoglobulin light chain, but negative for CD20, CD30, CD79a, PAX-5, CD10, and lambda light chain. In addition, the neoplastic cells showed positive immunoreactivity for anaplastic lymphoma kinase (ALK) with exclusive cytoplasmic granular staining pattern. Furthermore, interphase fluorescence in situ hybridization analysis, revealed ALK gene rearrangement in the lymphoid cells. The diagnosis of this rare entity in unusual extranodal sites such as the bone marrow is challenging, and it is based on detailed morphological and immunohistochemical analysis. Fluorescence in situ hybridization provides also a helpful tool in order to establish the diagnosis.