Browsing by Author "Vollrath, Valeska"

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    Analysis of natural variants of the hepatitis C virus internal ribosome entry site reveals that primary sequence plays a key role in cap-independent translation
    (OXFORD UNIV PRESS, 2009) Ines Barria, Maria; Gonzalez, Angel; Vera Otarola, Jorge; Leon, Ursula; Vollrath, Valeska; Marsac, Delphine; Monasterio, Octavio; Perez Acle, Tomas; Soza, Alejandro; Lopez Lastra, Marcelo
    The HCV internal ribosome entry site (IRES) spans a region of similar to 340 nt that encompasses most of the 5' untranslated region (5'UTR) of the viral mRNA and the first 24-40 nt of the core-coding region. To investigate the implication of altering the primary sequence of the 5'UTR on IRES activity, naturally occurring variants of the 5'UTR were isolated from clinical samples and analyzed. The impact of the identified mutations on translation was evaluated in the context of RLuc/FLuc bicistronic RNAs. Results show that depending on their location within the RNA structure, these naturally occurring mutations cause a range of effects on IRES activity. However, mutations within subdomain IIId hinder HCV IRES-mediated translation. In an attempt to explain these data, the dynamic behavior of the subdomain IIId was analyzed by means of molecular dynamics (MD) simulations. Despite the loss of function, MD simulations predicted that mutant G266A/G268U possesses a structure similar to the wt-RNA. This prediction was validated by analyzing the secondary structure of the isolated IIId RNAs by circular dichroism spectroscopy in the presence or absence of Mg2+ ions. These data strongly suggest that the primary sequence of subdomain IIId plays a key role in HCV IRES-mediated translation.
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    Bucillamine induces glutathione biosynthesis via activation of the transcription factor Nrf2
    (PERGAMON-ELSEVIER SCIENCE LTD, 2006) Wielandt, Ana M.; Vollrath, Valeska; Farias, Marcelo; Chianale, Jose
    The properties of bucillamine, a synthetic antioxidant, have been attributed mainly to the donation of thiol groups to glutathione (GSH). We recently demonstrated that glutamatecysteine ligase catalytic subunit (GCLC), the rate-limiting enzyme of GSH biosynthesis, and the multidrug-resistance-associated protein 2 (Mrp2/MRP2) are coordinately induced in response to xenobiotic through the activation of the antioxidant-response element (ARE) by nuclear factor-erythroid 2 p45-related factor (Nrf2). We tested the hypothesis that bucillamine and its oxidized metabolite SA 981 also activate the Nrf2 pathway, thereby increasing glutathione biosynthesis in human HepG2 and murine Hepa 1-6 hepatoma cell lines, through the induction of the GCLC enzyme as well as the Mrp2/MRP2 transporter, which mediates the excretion of glutathione and its conjugates from hepatocytes. Both bucillamine and SA 981 produced a significant dose-dependent increase in the mRNA levels of Mrp2/MRP2 and GCLC after 24 h. The levels of the transcription factor Nrf2 in the nuclei were maximal at 3 h, remained elevated at 6 h, and decreased to control values at 24 h in both cell lines. Moreover, both bucillamine and SA 981 significantly increased the expressions of Mrp2/MRP2 and GCLC proteins in both cell lines. Finally, in both cell lines, bucillamine and SA 981 increased the GSH content two- to three-fold. These results demonstrate that bucillamine and SA 981 activate the ARE-ARE pathway increasing the expression of ARE-driven genes such as those of GCLC and Mrp2/MRP2. The role of bucillamine as a chemopreventive agent against cancer remains to be elucidated. (c) 2006 Elsevier Inc. All rights reserved.
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    C(-106)T polymorphism of the aldose reductase gene and the progression rate of diabetic retinopathy
    (ELSEVIER IRELAND LTD, 2006) Olmos, Pablo; Bastias, Maria Juliana; Vollrath, Valeska; Toro, Luis; Trincado, Arturo; Salinas, Pablo; Claro, Juan Carlos; Lopez, Jose Manuel; Acosta, Ana Maria; Miquel, Juan Francisco; Castro, Juan
    Purpose: To study the C(-106)T polymorphism in the promoter of the aldose reductase (ALR2) gene: (a) its local prevalence and (b) its modulation of the susceptibility for developing retinopathy.
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    Influence of extrahepatic viral infection on the natural history of hepatitis C
    (ELSEVIER ESPANA, 2008) Ines Barria, Maria; Vera Otarola, Jorge; Leon, Ursula; Vollrath, Valeska; Marsac, Delphine; Riquelme, Arnoldo; Lopez Lastra, Marcelo; Soza, Alejandro
    HCV is primarily hepatotropic, but there is mounting evidence pointing to infection and replication of extrahepatic sites. Here we evaluated the occurrence of HCV infection of peripheral blood mononuclear cells (PBMC) and explored the possible association between viral extrahepatic infection and the natural history of the disease. Forty seven Chilean, HCV infected, treatment naive patients were included in the study. HCV RNA was isolated from plasma and PBMC and subsequently reverse transcribed, amplified and sequenced. Most patients harbored HCV 1b genotype and the most common route of infection showed to be blood transfusion. HCV RNA was readily detected in PBMCs of 34 out of the 47 patients (72%). We report that HCV sequences found in PBMC differ from those in plasma of the same subjects strongly suggesting HCV compartmentalization. In addition, we found that patients with detectable HCV RNA in PBMC had a tendency for being more likely cirrhotic [OR 3.8 (95 % CI: 0.98 to 14)]. In conclusion, this study provides further arguments for the existence of HCV infection of extrahepatic sites and suggests that extrahepatic infection could be a factor influencing the natural history of the disease.
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    Phenotype and genotype of thiopurine methyltransferase in Chilean individuals
    (SOC MEDICA SANTIAGO, 2012) Jorquera, Andres; Solari, Sandra; Vollrath, Valeska; Guerra, Irene; Chianale, Jose; Cofre, Colomba; Kalergis, Alexis; Ibanez, Patricio; Bueno, Susan; Alvarez Lobos, Manuel
    Background: Thiopurines (azathioprine and 6-mercaptopurine) are highly effective medications but with potential adverse effects. Thiopurine methyltransferase (TMPT) is the key enzyme in their pharmacokinetics and is genetically regulated. A low activity of TPMT is associated with myelotoxicity. The genotype and enzyme activity can vary by ethnicity. Aim: To study the activity and genotype of TPMT in a group of Chilean subjects. Material and Methods: In 200 healthy adult blood donors, TPMT activity was determined by high performance liquid chromatography (HPLC). Deficient, low, normal or high levels were defined when enzymatic activity was <= 5, 6-24, 25-55 and >= 56 nmol/grHb/h, respectively. Genotyping of TPMT ((star)1, (star)2, (star)3A, (star)3B, (star)3C) was performed by PCR. Results: Seventy seven women (38.5%) and 123 men (61.5%), with an average age of 34.9 years were studied. Eighteen subjects (9%) had a low enzymatic activity, 178 (89%) had normal activity, 4 (2%) had high activity and no genotype deficient subjects were identified. The wild type genotype ((star)1) was found in 184 (92%) individuals and 16 (8%) were heterozygous for the variants: (star)2 (n = 2), (star)3A (n = 13) and (star)3C (n = 1). No homozygous subjects for these variants were identified. Wild type genotype had an increased enzymatic activity (40.8 +/- 7.2 nmol/gHb/h) compared to heterozygous group (21.2 +/- 3 nmol/gHb/h; p < 0.001). Conclusions: Less than 10% of a Chilean population sample has a low enzymatic activity or allelic variants in the TPMT gene, supporting the use of thiopurines according to international recommendations. (Rev Med Chile 2012; 140: 889-895).