Prognostic Value of Association of Copy Number Alterations and Cell-Surface Expression Markers in Newly Diagnosed Multiple Myeloma Patients.

Multiple myeloma results from the clonal proliferation of abnormal plasma cells (PCs) in the bone marrow (BM). In this study, the cell surface expression markers (CD) on atypical PCs (detected by multiparametric flow cytometry (MFC)) were correlated with copy number alterations (CNAs) in the genome (detected by multiplex ligation-dependent probe amplification (MLPA)) to assess their impact on prognosis in newly diagnosed MM patients. Statistically significant results were obtained when different stages of PC maturation (classified based on CD19 and CD81 expression) were associated with CD117 expression and identified CNAs. In the intermediately differentiated PC group (CD19(-) CD81(+)), patients who didn't express CD117 had a lower median progression free survival (PFS) (p = 0.024). Moreover, within this group, patients with less than three adverse CNAs, which harbor CD117, had a better outcome with a PFS of more than 48 months compared with 19 months (p = 0.008). Considering all the results, our study suggested the need to integrate both the CD markers and copy number alterations to evaluate the prognosis of newly diagnosed multiple myeloma patients.

Modulation of the activity of certain genes involved in tumor cell metabolism in the presence of the cytotoxic peptides defensin and cathelicidin LL37.

It is common knowledge that some natural antimicrobial peptides also have a tumoricidal effect. We have shown that the peptides defensin and cathelicidin LL37 have cytostatic effects on human tumor cell lines HT29 (colorectal carcinoma) and A549 (alveolar carcinoma). In order to determine the modulating mechanism of these peptides we assessed the gene expression of the AKT, HIF-1α, XBP, NRF2, PERK, CHOP, BCL2, IRE1α and PI3K molecular targets involved in the survival, growth, proliferation and apoptosis pathways of tumor cells in the presence or absence of the studied peptides. Thus, this research enabled us to determine molecular markers and methods of assessment and monitoring of tumor cell cytotoxicity by high-performance molecular biology techniques. Defensin and cathelicidin LL37 activated tumor cell apoptosis, especially for the HT29, but also for A549 line, by increasing gene expression of CHOP and by lowering BCL2 gene expression. Oxidative stress determined the increase in gene expression of XBP, which directly influenced CHOP. The decrease in NRF2 gene expression highlighted the inhibition of cell proliferation, while the decrease in HIF1α gene expression evidenced the decrease in cell survival.

Inflammatory and Genetic Markers (APO B100 and Angiotensin-Converting Enzyme Gene) in the Coronary Artery Disease.

Aim: To analyze the correlations between inflammation markers and ApoB100 and angiotensin converting enzyme (ACE) gene polymorphism and the severity of coronary artery disease (CAD). Material and Methods: We conducted a study in 58 patients with acute coronary syndromes (ACS) who underwent coronarography at the Iasi "Prof. Dr. George I.M. Georgescu' Institute of Cardiovascular Diseases. the patients included in the studies were selected from those who needed a coronarography for unstable angina or acute myocardial infarction. The data were uploaded and processed using the statistical functions in SPPS 18.0 at a 95% materiality threshold. Results: Elevated inflammation markers were found in all study patients, with small differences in distribution. None of the study patients presented ApoB100 gene mutations. As to ACE polymorphism, a predominance of genotype II in unicoronary patients and ID and DD genotypes in bicoronary and tricoronary patients was found. Conclusions: The results of our study confirm the role of genetic and epigenetic factors in the severity and progression of the coronary disease, leaving room for larger and more comprehensive studies and new research perspectives.

The cytotoxic effect of magainin II on the MDA-MB-231 and M14K tumour cell lines.

Many studies have highlighted the tumoricidal properties of some natural peptides known to have antimicrobial virtues. Also, the increasingly higher resistance to conventional antibiotics has become a global public health issue, and the need for new antibiotics has stimulated interest in finding and synthesizing new antimicrobial peptides, which may also be used as chemotherapeutic agents. Relying on the literature, the purpose of our in vitro research was to assess the tumoricidal potential of magainin II on a series of tumour cell lines, namely, MDA-MB-231 (breast adenocarcinoma) and M14K (human mesothelioma). The experimental results of our study revealed that the cytotoxic effects of magainin II depend on its concentration. Its efficiency is significant at 120 µM concentrations, and, although it is much lower, it persists even at 60 µM concentrations. The effects were insignificant at 30 µM concentrations. In our experimental research, the tumoricidal effect of magainin II was not significantly dependent on the type of tumour cell line used.

Digital microscopy assessment of angiogenesis in different breast cancer compartments.

BACKGROUND/AIM: Tumour angiogenesis defined by microvessel density (MVD) is generally accepted as a prognostic factor in breast cancer. However, due to variability of measurement systems and cutoffs, it is questionable to date whether it contributes to predictive outline. Our study aims to grade vascular heterogeneity by comparing clear-cut compartments: tumour associated stroma (TAS), tumour parenchyma, and tumour invasive front. MATERIAL AND METHODS: Computerized vessel area measurement was performed using a tissue cytometry system (TissueFAXS) on slides originated from 50 patients with breast cancer. Vessels were marked using immunohistochemistry with CD34. Regions of interest were manually defined for each tumour compartment. RESULTS: Tumour invasive front vascular endothelia area was 2.15 times higher than that in tumour parenchyma and 4.61 times higher than that in TAS (P < 0.002). Worth to mention that the lymph node negative subgroup of patients show a slight but constant increase of vessel index in all examined compartments of breast tumour. CONCLUSION: Whole slide digital examination and region of interest (ROI) analysis are a valuable tool in scoring angiogenesis markers and disclosing their prognostic capacity. Our study reveals compartments' variability of vessel density inside the tumour and highlights the propensity of invasive front to associate an active process of angiogenesis with potential implications in adjuvant therapy.

Screening for latent BK virus infection in a renal transplant population for the first time in Romania: a single-center experience.

INTRODUCTION: The BK virus nephropathy (BKVN) is one of the most important infectious complications in renal transplant recipients. As BKVN lacks any effective antiviral treatment, early diagnosis is required in order to try to limit viral replication and subsequent damage to the renal allograft, by reducing the immunosuppressive therapy. Our study, the first of its kind in Romania, aimed to assess the prevalence of BKVN among renal transplant patients in our center. MATERIALS AND METHODS: In this cross-sectional study, we included 143 renal transplant patients from our center who had received their renal allograft between 2005 and 2010. We searched for latent BK virus infection by detection of serum anti-BK virus antibodies, using an in-house developed enzyme-linked immunosorbent assay (ELISA) technique. Serology was considered positive if results were >0.33 optical density units. In patients with positive serology, we searched for BKVN with qualitative (polymerase chain reaction, PCR) and quantitative (TaqMan real-time PCR) molecular techniques. Additionally, we searched for other viral infections, including hepatitis B (with HBsAg test), hepatitis C (with anti-HCV Abs test), and cytomegalovirus (CMV, with pp65Ag test). RESULTS: All patients screened with ELISA were found to have positive BK virus serology and two of these were diagnosed with BKVN. Both patients with BKVN presented with acute impairment of the renal graft function, and one of them also developed a ureteral graft stenosis. In both cases, BKVN resolved after reduction of immunosuppressive doses. We also diagnosed hepatitis B in 18.18%, hepatitis C in 7.0%, and CMV in 27.97% of patients. CONCLUSIONS: Our study demonstrates for the first time the existence of BK virus in Romania, and we believe it opens the prospective of diagnosing BKVN in high-risk patients in our country in the future. In renal transplant patients from our center, we found the prevalence of BK virus infection to be as high as 100%. The prevalence of hepatitis B and CMV was also remarkably high. In patients with BKVN, the reduction of immunosuppression enables the spontaneous resolution of the disease.

[Prognostic significance of leukemia-associated phenotype in correlation with other biologic markers in acute myeloid leukemia patients]. / Identificarea semnificatiei prognostice a fenotipului asociat leucemiei in corelatie cu alti markeri biologici la pacientii cu leucemie acuta mieloblastica.

UNLABELLED: Leukemic cells have unique aberrant phenotypes, which permit identification of this cells at diagnose and in evolution of the disease. Signaling molecules with other cells and bone marrow stroma are part of the leukemic cells phenotype. Genetic and molecular abnormalities have the main prognostic significance and confer the leukemic cell status. The main aim of the current study is to identify correlation between recognized prognostic factors in acute myeloid leukemia (AML) patients and other phenotypic markers. MATERIAL AND METHOD: Imunophenotypic analysis (BDFACS CantoII, FACSDiva Software) was performed on peripheral blood/bone marrow aspirate samples of 56 patients diagnosed with AML (9 M0, 3 M1, 10 M2, 4 M3, 28 M4/M5, 1 M6, 1 M7) between 2007-2009 in Hematology Department of "Sf. Spiridon" Hospital Iasi. We used an extended panel of monoclonal antibodies and we determined the level of expression of cytokines receptors (IL3Ra, IL7R) and chemokines (CXCR4, CKR5). RESULTS: In our study, IL7R expression on AML blasts was significant correlate with low WBC count at diagnosis (p = 0.04) and with multilinear displasia (p = 0.01), high CXCR4 expression was correlate (p = 0.05) with lack of response at first induction therapy and CD123 (IL3Ra) expression was correlate with M4 FAB phenotype. Survival was negative influenced by presence of IL3R on AML blasts, but flt3 mutations, CXCR4, IL7R expression on leukemic cells, other phenotypic aberrancies did not influenced treatment response and survival in our patients population. CONCLUSION: Complete investigation of leukemic cells phenotype extended with cytokines/chemokines receptors at diagnostic is useful for correct characterization of the disease, for discover new prognostic categories and for better identification of minimal residual disease.