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Papers para osvaldo

A simple allele-specific polymerase chain reaction method to detect the
Gly143Glu polymorphism in the human carboxylesterase 1 gene:
importance of genotyping for pharmacogenetic treatment.

Walter Soria N, Belaus A, Galván C, Ana Pasquali M, Velez P, Del Carmen Montes C,
Beltramo DM.
Cátedra de Biotecnología, Facultad de Ciencias Químicas, Universidad Católica de
Córdoba, Córdoba, Argentina.
Genet Test Mol Biomarkers. 2010 Dec;14(6):749-51. Epub 2010 Sep 21.
Human carboxylesterases 1 and 2 (CES1 and CES2) catalyze the hydrolysis of many exogenous compounds. Alterations in CES sequences could lead to variability in both the inactivation of drugs and the activation of prodrugs. The human CES1 gene encodes for the enzyme carboxylesterase 1, a serine esterase governing both metabolic deactivation and activation of numerous therapeutic agents. Some of theses drugs are the antiviral oseltamivir used to treat some types of influenza infections and the methylphenidate employed in the treatment of patients with attention deficit. The Gly143Glu polymorphism in CES1 gene has been shown to reduce enzyme activity. The aim of the present study was to develop an easy and cheap method to detect this polymorphism. For this, we studied a group of people from Córdoba, a Mediterranean area from Argentina. Our results show that our methodology could detect the presence of this polymorphism with a frequency around 1.8%, only in the heterozygote form. These results could be relevant to patients before the treatment with some drugs where the CES1 enzyme is involved. Rapid HCP5 single-nucleotide polymorphism genotyping: a simple
allele-specific PCR method for prediction of hypersensitivity reaction
to Abacavir.

Galván CA, Elbarcha OC, Fernández EJ, Beltramo DM, Soria NW.
Laboratorio de Análisis Clínicos Especializados (LACE), Córdoba, Argentina.
Clin Chim Acta. 2011 Jul 15;412(15-16):1382-4. Epub 2011 Apr 14.
BACKGROUND: The most important factor limiting the success of an antiretroviral
therapy is toxicity. The HLA-B*5701 allele is predictive of hypersensitivity reaction to
Abacavir, and this gene is in a perfect linkage disequilibrium with the rs2395029 SNP
present in the HCP5 gene.
METHODS: Genomic DNA was extracted from blood obtained from 201 unrelated
healthy Argentinean volunteers. The DNA was subjected to an allele-specific PCR
method. Sequencing was performed to validate the test results.
RESULTS: We were successful to amplify specific fragment of interest from the DNA
samples. The method is easy, specific and reproducible.
CONCLUSIONS: The application of this methodology is a rapid and simple method to
detect the HCP5 polymorphism (rs2395029) previous to administration of Abacavir in
patients with HIV infection.
Genetic profiling of GSTP1, DPYD, FCGR2A, FCGR3A and CCND1
genes in an Argentinian population.

Galván CA, Elbarcha OC, Fernández EJ, Beltramo DM, Soria NW.
Laboratorio de Análisis Clínicos Especializados (LACE), Córdoba, Argentina.
Biochem. Abstract
OBJECTIVES: To determine the frequencies of relevant allelic variants in oncology
for the GSTP1, DPYD, FCGR2A, FCGR3A and CCND1 genes in a population from
Central Argentina. To compare the allelic distribution found with the frequencies
reported for other ethnic groups.
DESIGN AND METHODS: Genotyping was carried out in a total of 102 unrelated
Argentinian subjects. FCGR3A (rs396991) was detected using allele specific
polymerase chain reaction (PCR) assay, while GSTP1 (rs1695), DPYD (rs3918290),
FCGR2A (rs1801274) and CCND1 (rs9344) variants were assessed by PCR-restriction
fragment length polymorphism (PCR-RFLP).
RESULTS: The allele frequencies for GSTP*1B, DPYD*2A, FCGR2A (131R),
FCGR3A (158F) and CCND1 (870G) in Argentinians were 0.35, 0.005, 0.41, 0.77 and
0.47, respectively.
CONCLUSIONS: We found that the Argentinian population tested resembles other
Caucasians populations, especially Spaniards; yet the differences in allele distribution
with other Caucasian groups, uncover population admixture with native Amerindian and
other ethnic groups, consistent with the well documented immigration flows landing
Argentina from several countries.
Development of a method to control the RNA extraction and reverse
transcription steps for the detection of dengue virus present in human
blood samples.

Galván CA, Elbarcha OC, Fernández EJ, Beltramo DM, Soria NW.
Laboratorio de Análisis Clínicos Especializados (LACE), Córdoba, Argentina.
Genet Test Mol Biomarkers. 2011 Dec;15(12):913-5. Epub 2011 Jun 20.
AIMS: Molecular biology techniques based on the detection of genomic sequences by
reverse transcription combined with polymerase chain reaction (PCR) have enabled the
detection of different RNA viruses in serum or plasma samples. Since the dengue
epidemic outbreak declared in Argentina in 2009, numerous patients' samples were
analyzed for the acute phase of infection. One of the main methodological drawbacks is
the lack of internal control to measure the effectiveness of the viral extraction and
reverse transcription process. In this article, we propose to standardize a molecular
method to detect beta actin (β-Act) and glucose 6 phosphate dehydrogenase (G6PDH)
complementary DNAs (cDNAs) present in patient's plasma/serum, as a control process.
RESULTS: RNA extraction, reverse transcription, and PCRs for human G6PDH, β-
Act, and the dengue virus genome were performed. cDNA fragments for β-Act and
G6PDH were amplified for all samples, regardless of the presence or absence of viral
CONCLUSIONS: Amplification of β-Act and G6PDH cDNAs can be used as a control
for the extraction and reverse transcription processes during dengue virus detection.
This could also be a useful method for controlling the above steps when infections
caused by other RNA viruses are studied, even if another methodology is employed,
such as real-time PCR.

Human Immunodefieciency Virus: Pharmacogenetics of Antiretroviral
Susana A Pesoa1, Cristian A Galván1,2, Dante M Beltramo2,3,4 and Néstor W Soria1,2
1Laboratorio de Análisis Clínicos Especializados (LACE), Córdoba, Argentina
2Cátedra de Biotecnología, Facultad de Ciencias Químicas, Universidad Católica de
Córdoba, Córdoba, Argentina
3Centro de Excelencia en Productos y Procesos de Córdoba (CEPROCOR), Santa
María de Punilla, Córdoba, Argentina
4Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)
Pharmacogenomics & Pharmacoproteomics.
Pesoa et al., J Pharmacogenom
Pharmacoproteomics 2011, S6

The introduction of highly active antiretroviral therapy as standard of care has
considerably enhanced the life expectancy among HIV-infected individuals. Although
this combination of drugs virtually suppresses viral replication, the therapeutic effect
may be limited by differing rates of adverse events and responses in terms of efficacy
and toxicity. These differences arise from complex interactions between biological and
environmental factors. Pharmacogenetic studies are contributing to our understanding of
the inter-individual differences in the response to antirretroviral drugs. Several studies
have provided a relevant number of associations between human genetic variants and
predisposition to adverse events and for some antiretroviral drugs clear and causal
genotype–phenotype correlation has been established. These findings make the idea of
personalized medicine in this field increasingly attractive. We will discuss here current
achievements on pharmacogenetics of HIV treatment with special emphasis on the
genetic polymorphisms underlying toxic effects and/or those already implemented in
the clinical setting
histocompatibility complex P5, chemokine coreceptor 5, and
interleukin 28B genes in inhabitants from the central area of

Galván CA, Elbarcha OC, Fernández EJ, Beltramo DM, Soria NW.
Laboratorio de Análisis Clínicos Especializados (LACE), Córdoba, Argentina.
Genet Test Mol Biomarkers. 2012 Feb;16(2):130-3. Epub 2011 Aug 19.
AIMS: The selection of the most appropriate treatment for several diseases relies on a
number of factors such as environment, age, gender, and nutrition. Additionally, the
contribution of different genetic polymorphisms to treatment efficacy has been largely
recognized. The lack of information on the pharmacogenetic profile of our population
prompted us to analyze the frequency of polymorphisms known to be relevant to
achieve treatment efficacy with different therapeutic agents in viral infectious diseases,
such as Hepatitis C and AIDS.
RESULTS: The allelic frequencies for the wild-type variant of the genes analyzed were
cytochrome P450 2B6 (CYP2B6; rs3745274; 516G) 0.618 (95% confidence interval
[CI]: 0.523, 0.711), chemokine coreceptor 5 (CCR5; rs333) 0.961 (95% CI: 0.942,
0.98), histocompatibility complex P5 (HCP5; rs2395029; 335T) 0.971 (95% CI: 0.937,
1), and interleukin 28B (IL28B; rs12979860; 12007005C) 0.656 (95% CI: 0.564,
0.747), respectively.
CONCLUSIONS: Our data indicate that the genetic profile of the population studied is
similar to that reported for other Caucasian populations, with only slight differences for
CYP2B6. Noteworthy, the considerable number of patients carrying CYP2B6 (516T)
and IL28B (12007005T) alleles underlies the importance of considering
pharmacogenetic testing before starting drug therapy protocols to prevent toxicity
and/or lack of effectiveness in AIDS or hepatitis C virus infections.


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