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Marília, Brazil

Viana-Baracioli L.M.S.,University Estadual Of Sao Paulo Julio Of Mesquita Filho | Junior N.C.T.,University Estadual Of Sao Paulo Julio Of Mesquita Filho | Junior O.R.,Hemocentro | Angulo I.L.,University of Sao Paulo | Bonini-Domingos C.R.,University Estadual Of Sao Paulo Julio Of Mesquita Filho
Genetics and Molecular Research

It is well documented that Hb S and iron affect blood cells, and trigger oxidative processes and generation of free radicals with potential for lipid peroxidation. We evaluated the frequency of polymorphisms in the HFE gene in Hb AS blood donors and how these polymorphisms influenced lipid peroxidation and antioxidant capacity. Blood samples were collected from 211 Hb AS blood donors, 119 Hb AA blood donors as a control group, and 28 sickle cell disease patients (Hb SS). The H63D allele was found at a frequency of 10.5% in the Hb AS samples, and the C282Y allele frequency was 0.7%. In the control group, the frequencies of the H63D and C282Y alleles were 13.4 and 2.1%, respectively. In the sickle-cell disease patients, the H63D and the C282Y allele frequencies were 10.7 and 3.5%, respectively. The frequencies of the C282Y and H63D polymorphisms in Hb AS blood donors are similar to those reported for the Brazilian population. Serum malondialdehyde values, indicative of lipid peroxidation, were highest in sickle cell patients, independent of the polymorphisms in the HFE gene, with significant differences, showing the influence of Hb S allele in the levels of lipid peroxidation. However, the trolox equivalent antioxidant capacity average levels, indicative of the antioxidant capacity, were reduced with significant differences, indicating that in spite of a lipid peroxidation raise, this is not followed by the increased of the antioxidant capacity, leading to oxidative stress. © FUNPEC-RP www.funpecrp.com.br. Source

Shimauti E.L.T.,Sao Paulo State University | Shimauti E.L.T.,State University of Maringa | Silva D.G.H.,Sao Paulo State University | De Souza E.M.,State University of Maringa | And 3 more authors.
Genetics and Molecular Biology

The aim of this study was to determine the frequency of beta S-globin gene (βS globin) haplotypes and alpha thalassemia with 3.7 kb deletion (-α3.7kb thalassemia) in the northwest region of Paraná state, and to investigate the oxidative and clinical-hematological profile of βS globin carriers in this population. Of the 77 samples analyzed, 17 were Hb SS, 30 were Hb AS and 30 were Hb AA. The βS globin haplotypes and - α3.7kb thalassemia were identified using polymerase chain reaction.Trolox equivalent antioxidant capacity (TEAC) and lipid peroxidation (LPO) were assessed spectophotometrically. Serum melatonin levels were determined using high-performance liquid chromatography coupled to coulometric electrochemical detection. The haplotype frequencies in the SS individuals were as follows: Bantu- 21 (62%), Benin - 11 (32%) and Atypical- 2 (6%). Bantu/Benin was the most frequent genotype. Of the 47 SS and AS individuals assessed, 17% (n = 8) had the - α3.7kb mutation. Clinical manifestations, as well as serum melatonin, TEAC and LPO levels did not differ between Bantu/Bantu and Bantu/Benin individuals (p > 0.05). Both genotypes were associated with high LPO and TEAC levels and decreased melatonin concentration. These data suggest that the level of oxidative stress in patients with Bantu/Bantu and Bantu/Benin genotypes may overload the antioxidant capacity. © 2015, Sociedade Brasileira de Genética. Source

Wisnieski F.,University of Sao Paulo | Calcagno D.Q.,University of Sao Paulo | Calcagno D.Q.,Federal University of Para | Leal M.F.,University of Sao Paulo | And 12 more authors.
Tumor Biology

Gastric cancer is still the second leading cause of cancer-related death worldwide, even though its incidence and mortality have declined over the recent few decades. Epigenetic control using histone deacetylase inhibitors, such as trichostatin A (TSA), is a promising cancer therapy. This study aimed to assess the messenger RNA (mRNA) levels of three histone deacetylases (HDAC1, HDAC2, and HDAC3), two histone acetyltransferases (GCN5 and PCAF), and two possible targets of these histone modifiers (MYC and CDKN1A) in 50 matched pairs of gastric tumors and corresponding adjacent nontumors samples from patients with gastric adenocarcinoma, as well as their correlations and their possible associations with clinicopathological features. Additionally, we evaluated whether these genes are sensitive to TSA in gastric cancer cell lines. Our results demonstrated downregulation of HDAC1, PCAF, and CDKN1A in gastric tumors compared with adjacent nontumors (P<0.05). On the other hand, upregulation of HDAC2, GCN5, and MYC was observed in gastric tumors compared with adjacent nontumors (P<0.05). The mRNA level of MYC was correlated to HDAC3 and GCN5 (P<0.05), whereas CDKN1A was correlated to HDAC1 and GCN5 (P<0.05 and P<0.01, respectively). In addition, the reduced expression of PCAF was associated with intestinal-type gastric cancer (P=0.03) and TNM stages I/II (P=0.01). The increased expression of GCN5 was associated with advanced stage gastric cancer (P=0.02) and tumor invasion (P=0.03). The gastric cell lines treated with TSA showed different patterns of histone deacetylase and acetyltransferase mRNA expression, downregulation of MYC, and upregulation of CDKN1A. Our findings suggest that alteration of histone modifier genes play an important role in gastric carcinogenesis, contributing to MYC and CDKN1A deregulation. In addition, all genes studied here are modulated by TSA, although this modulation appears to be dependent of the genetic background of the cell line. © 2014 International Society of Oncology and BioMarkers (ISOBM). Source

Baleotti W.,Hemocentro | Suzuki R.B.,Hemocentro | Polotto M.,FAMERP | Ruiz M.O.,Hemocentro | And 2 more authors.
Journal of Clinical Laboratory Analysis

Determination of the molecular basis underlying the antigens in the Dombrock blood group system has shown various rearrangements between the alleles associated with DO z.ast;A and DO z.ast;B. Based on this, we employed a PCR-based strategy to screen DO alleles (DO z.ast;A, DO z.ast;B, HY z.ast;1, HY z.ast;2 and JO) in Brazilians. Methods: We tested DNA of 278 Brazilian blood donors by PCR-RFLP on plates of 96 wells to determine the 793A/G (DO z.ast;A/DO z.ast;B), 323G/T (HY), 350C/T (JO) and 898C/G (HY z.ast;1/HY z.ast;2) single nucletide polymorphisms. In order to confirm the results sequence analysis was also performed. Results: When samples of these donors were analyzed, a novel allele combination, the DO z.ast;A allele (793A and 323G) associated with 898G was identified and designated as DO z.ast;A-WL allele. This new allele encoding 300Val is the same as HY z.ast;1 at nucleotide 898 on the molecular background of DO z.ast;A. Among the 556 alleles analyzed by PCR-RFLP, 3 were DO z.ast;A-WL and 78 were DO z.ast;B-WL. This represents an overall frequency of 0.5% for DO z.ast;A-WL and 14% for DO z.ast;B-WL across the population studied. Conclusion: Molecular screening of Brazilians revealed one novel allele, the DO z.ast;A-WL. Our data highlight the importance of testing a cohort of different populations to determine DO haplotypes and to establish reliable genotyping tests for predicting Do a/Do b status. © 2011 Wiley-Liss, Inc. Source

Ferraz M.A.,Sacred Heart University of Brazil | Zabaglia L.M.,Sacred Heart University of Brazil | Pereira W.N.,Sacred Heart University of Brazil | Orcini W.A.,Sacred Heart University of Brazil | And 11 more authors.
Journal of Gastrointestinal Cancer

Purpose: Gastritis caused by infection with Helicobacter pylori is characterized by chronic inflammation and damage in gastric tissue, which is a main risk factor for gastric cancer. Associated with H. pylori, the TP53 gene tumor suppressor and the cell adhesion glycoprotein epithelial cadherin develop a relevant role in the integrity and carcinogenesis of the epithelium. We aimed to detection of H. pylori and its main virulence markers and measured the messenger RNA (mRNA) expression levels of E-cadherin and TP53 genes. Methods: The detection of H. pylori and its virulence markers, as well as the mRNA expression levels of E-cadherin and TP53 genes, were obtained for 161 samples of gastric biopsies including 37 with normal gastric tissue, 70 with gastritis, 24 from neoplastic tissue, and 27 from adjacent non-neoplastic by means of a quantitative real-time polymerase chain reaction. Results: The mRNA expression levels of E-cadherin and TP53 were found to be decreased in patients with gastritis, independently of H. pylori infection. In samples from gastric patients, the neoplastic tissue showed an accentuated decrease of expression; on the other hand, the expression of E-cadherin was normal in adjacent non-neoplastic. Conclusions: No evidence was found of the involvement of the cagA and vacA genes in the decreased expression of E-cadherin and TP53. The process of carcinogenesis is complex, and the decrease of the E-cadherin gene expression and TP53 gene expression appears to contribute significantly. © 2015, Springer Science+Business Media New York. Source

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