CytoregR inhibits growth and proliferation of human adenocarcinoma cells via induction of apoptosis.

BACKGROUND: Cancer is one of the devastating neovascular diseases that incapacitate so many people the world over. Recent reports from the National Cancer Institute indicate some significant gain therapy and cancer management as seen in the increase in the 5-year survival rate over the past two decades. Although near-perfect cure rate have been reported in the early-stage disease, these data reveal high recurrence rate and serious side effects including second malignancies and fatalities. Most of the currently used anticancer agents are only effective against proliferating cancer cells. Thus attention has been focused on potential anti-cancer agents capable of killing cancer cells independent of the cell cycle state, to ensure effective elimination of most cancer cells. The objective of this study was to test the chemosensitivity and potential mechanism of action of a novel cancer drug, CytoregR, in a panel of human cancer cells. METHODS: the study was performed using a series of bioassays including Trypan blue exclusion, MTS Growth inhibition, LDH-cytotoxicity, TUNEL-Terminal DNA fragmentation Apoptosis Assay, and the Caspase protease CPP32 activity assays. RESULTS: CytoregR induced significant dose- and time-dependent inhibition of growth in all the cells; with significant differences in chemosensitivity (P < 0.05) between the target cells becoming more apparent at 48 hr exposure. CytoregR showed no significant effect on normal cells relative to the tumor cells. Growth inhibition in all the cells was due to induction of apoptosis at lower concentrations of cytoregR (> 1:300). CytoregR-induced caspase protease-3 (CPP32) activation significantly and positively correlated with apoptosis induction and growth inhibition; thus implicating CPP32 as the principal death pathway in cytoregR-induced apoptosis. CONCLUSION: CytoregR exerted a dose-and time-dependent growth inhibitory effect in all the target cells through induction of apoptosis via the CPP32 death pathway, independent of hormonal sensitivity of the cells. The present data indicate that not only could CPP32 provide a potential target for regulation of cytoregR-induced apoptosis but also that cytoregR could play a significant role in chemotherapeutic regimen in many human malignant tumors.

Influence of genistein isoflavone on matrix metalloproteinase-2 expression in prostate cancer cells.

We investigated the expression of matrix metalloproteinase (MMP)-2 in human LNCaP and PC3 prostate cancer cell lines in response to genistein exposure. Initially we studied the phytosensitivity of the cells to genistein using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay to determine percentage cell viability/inhibition and the terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end-labeling apoptosis assay to assess the type of cell death. The results revealed that genistein inhibited growth and proliferation in both PC3 (hormone-dependent) and LNCaP (hormone-independent) prostate cancer cell lines, that there was no significant difference in sensitivity to genistein between PC3 and LNCaP cells, and that the effect of genistein on the cells was dose- and time-dependent. The results also revealed that inhibition of cell growth in both PC3 and LNCaP cells was predominantly due to apoptotic cell death. These results were consistent with data in previous studies. This was followed by determination of the MMP-2 profile in response to genistein treatment. The results indicated a significant dose- and time-dependent inhibition of MMP-2 expression levels in both cells, with a highly significant negative correlation between MMP-2 levels and concentration of genistein. This is of phytotherapeutic significance in view of the pivotal role of MMP-2 expression in the pathogenesis of prostate cancer. Increasing expression of MMPs has been identified in many human cancers, including prostate cancer. Our findings indicate that genistein could be a potent therapeutic inhibitor of MMP-2 in line with current concepts of targeted treatment.

[Comparative study of indirect immunofluorescence (IIF) and ELISA techniques in the detection of parvovirus B19]. / Estudio comparativo de las técnicas de inmunofluorescencia indirecta (IFI) y ELISA para la detección de parvovirus B19.

To determine the seroprevalence of IgG and IgM antibodies against Parvovirus B19 (P. B19), we studied the sera of 53 patients with different hematologic disorders and the sera of 15 controls using indirect immunofluorescence (IFI) and the ELISA method. The prevalence of IgG in the control group was 46.6%, in patients with aplastic crisis was 83.3% (IFI) and 66.7% (ELISA) and, in patients without crisis was 68.9% (IFI) and 72.4% (ELISA). IgM was negative except for patients with crisis: 8.3% (IFI) and 29.1% (ELISA). The higher seroprevalence (IgG) found in patients in comparison with controls might be due to a greater exposure of of patients to the virus. The agreement for both techniques was 81%(IgG) and 93% (IgM) however ELISA technique was more sensitive for detecting IgM of P. B19. In spite of serologic evidence and evaluating a simple serum sample per patient, we could establish an association between aplastic crisis and viral infection for IgM ELISA but not for IgG between hematologic disorders and infection for the P. B19.

[Detection of parvovirus B19 in patients of the Zulia State blood bank with different hematological alterations]. / Detección de parvovirus B19 en pacientes del banco de sangre del Estado Zulia con diversas alteraciones hematológicas.

PURPOSE: To assess the seroprevalence of IgG and IgM specific for parvovirus B19 (B19) and its association with aplastic crisis developing in patients with different haematological disorders. PATIENTS AND METHODS: Fifty-three serum samples were evaluated, 24 from patients with aplastic crisis and 29 from others without such crises, all of them suffering from different haematological diseases diagnosed at the University Hospital of Maracaibo and the Zulia State Blood Bank, in Venezuela; 15 samples from healthy blood donors were examined as well. Indirect immunofluorescence technique was used in the study. Lymphocyte subsets were quantified in 10 of the patients with aplastic crisis by means of cytofluorometry. Serum proteins were assessed by electrophoresis in all samples. The statistical analysis was performed according to Student's t test and chi square, by applying the statix 4.0 and SAS programmes. RESULTS: Positive IgG was found in 20 of the 24 patients with aplastic crisis (83.3%), 20 of the 29 without crisis (68.9%) and 7 of the 15 healthy controls (46.6%). Positive IgM was found only in 2 of the 24 patients with aplastic crisis (8.3%). Both the patients without aplastic crises and the control groups were negative for PB19 IgM. The average CD4 and CD8 T lymphocyte count and the CD4-CD8 index in the patients studied were 454 CD4 cells/microL, 1,006 CD8 cells/microL and 0.5%, significantly different from the control group, whose figures were 860 CD4 cells/microL, 546 CD8 cells/microL and 1.6%. The average B lymphocyte count of the patients (628 cells/microL) was higher than that of the control group (349 cells/microL). The average NK cell count in the patient was 174 cells/microL, slightly below the control group (221 cells/microL). Mild beta-globulin decrease was found in the electrophoretic study of the serum proteins of the patients, along with significant increase of the total protein and the gammaglobulin fraction with regard to the control group. CONCLUSIONS: Higher PB19 IgG seropositivity was seen in the patients with aplastic crisis with respect to the control group, suggesting wider exposition to the virus among them with regard to the healthy population. Specific PB19 IgM was detected in 2 patients with immunodeficiency and aplastic crisis. A significant decrease of the CD4 and CD8 T-lymphocyte subsets, along with decreased CD4-CD8 quotient, were found in the aplastic crisis group, and an impairment of the immune response to the viral challenge can be inferred form this. The alterations of the serum proteins, together with the increased B lymphocytes, might suggest a polyclonal activation of these cells. The absence of other known lymphotropic viruses in most of the patients studied (50) show that the alterations found here are related to recent or past B19 infection.

Infección por virus linfotrópico T humano (VLTHI/II) en pacientes del banco de sangre e índigenas bari, Maracaibo, Estado Zulia, Venezuela / Infection by human T lymphotropic virus (HTLVI/II) in patients of the blood bank and Bari native indians, Maracaibo, State Zulia, Venezuela

La prueba de ELISA para la detección del virus linfotrópico T humano tipos I y II (VTLH-I/II) se prácticó en 230 muestras de sueros sanguíneos de donantes de sangre voluntarios, en 22 pacientes con leucemia y 52 con trastornos hematológicos, del banco de sangre del Estado Zulia. Se evaluaron 117 sueros de índigenas Bari de la Sierra de Perijá. Sólo 13 de las 421 muestras procesadas (3,08 por ciento) resultaron positivas para el VLTH-I/II. Sólo 1 suero de los 230 donates estudiados resultó positivo, representando un 0,4 por ciento. Los sueros de los pacientes leucémicos resultaron negativos. Un total de 8 de los 52 sueros de pacientes con trastornos hematológicos (15,4 por ciento) resultaron positivos. Por la otra parte, 4 de los 117 sueros provenientes de índigenas Bari (3,4 por ciento) mostraron positividad. En los pacientes con trastornos hematológicos se observó una mayor proporción de positivos en el grupo comprendido entre 20-29 años de edad. La proporción de sueros positivos en los pacientes del sexo femenino fue mayor. Los resultados de este estudio demostraron la presencia de una alta prevalencia a la infección por el VLTH-I/II en la población zuliana, los pacientes con leucemia deberán ser evaluados mediante pruebas de biología molecular con la finalidad de detectar la presencia de VLTH-I/II en las células

[Relation of spondylarthropathies and HLA-B27 antigen in patients from the state of Zulia, Venezuela]. / Estudio de la relación existente entre las espondiloartropatías y el antígeno HLA-B27 en pacientes del estado Zulia de Venezuela.

HLA-B27 and associate antigens incidence were studied in 620 cases of seronegative spondiloarthropathies (SNS) and 262 controls of a Venezuelan mestizo population from Zulla state between 1985 and 1995. The incidence of HLA-B27 was 20.96% of all cases of SNS. It was increased in patients with ankilosing spondylitis (AS) 33.33% and Relter's syndrome (RS) 30%, but not in uveitis (Uv) 20% an psoriatic arthropathy (PsA) 0%. The incidence in the control group was 4.2%. This results are lower than the previous description in Venezuelan mestizo and caucasic population but are close to the incidence described in population of West Africa with important contribution to admixture of the occidental part of Venezuela.

[Relationship between HLA antigens and the HIV infection in patients from the state of Zulia]. / Estudio de la relación entre antígenos HLA y la infección por VIH en pacientes del estado de Zulia.

PURPOSE: Some genetic factors associated to the HLA system phenotypes may allegedly predispose to the development of infection in patients exposed to the human immunodeficiency virus (HIV). So the aim of this study was to assess if certain HLA antigens are positive or negative risk factors in the development of AIDS in Zulia State. PATIENTS AND METHODS: A total of 62 samples were studied, 31 from HIV seropositive subjects and 31 form healthy individuals. The patients were subclassified into four groups in accordance with Atlanta's CDC guidelines. Tests for histocompatibility including HLA-A-B-C, DR and DQ typing were performed with Terasaki's technique. VIH positivity was determined by ELISA and confirmed by Western Blot. The statistical evalub1p4n was performed with the chi 2 test for antigen frequency comparison, the relative risk (RR) was estimated with the Ryder and Svelgaard test, and the inferential analysis was made by means of non-parametric statistics. RESULTS: Most patients were included in CDC's groups II and IV, 48.4% and 29.0%, respectively. Increased B35 and DQw2 and decreased B39 and DR2 antigens were found when comparing the HLA distribution in the sample and the antigenic frequency of the population. RR > 1 was observed in the infected patients A for A1, A3, A10, A11, B5, B7, B12, B14, B35, B61, CW4, DR4, DRW52 and DQW2 HLA antigens. A positive association between symptomatic infected patients and antigen B35 was present (X = 7.045). CONCLUSION: The findings reported here suggest that antigen B35 is a major risk factor for the development of AIDS.