Transcriptional study of genes involved in the passage from teliospore to hyphae stage in the fungus Thecaphora frezii, the causal agent of peanut smut.

Peanuts (Arachis hypogaea L.) are among the most important leguminous crops in Argentina. During the growing season, they are frequently attacked by fungal diseases, including Thecaphora frezii. The spores of T. frezii are structures that confer resistance to this phytopathogen. The transition from teliospore to hypha is a characteristic process of some fungi, which is essential for completing their life cycle. Using the transcriptomes of teliospores and hyphae of T. frezii, we aimed to identify genes that were differentially expressed during this transition, and we found 134 up-regulated and 66 down-regulated genes, which would participate in different cellular processes such as: (a) cell cycle and DNA processing; (b) cell fate; (c) rescue, defense and cellular virulence; (d) detoxification by CYP450; (e) energy; (f) nutrient interaction and nutritional adaptation; (g) metabolism; (g) proteins with binding functions or cofactor requirements; (h) stress, cell differentiation and biogenesis of cell components; and (i) transport, cell communication and transcription. The identification of genes in T. frezii and their expression levels during different stages of differentiation could contribute to our understanding of the biological mechanisms in this fungus.

Changes of lipids composition in different ontogenetic stages of Thecaphora frezii: expression of key enzymes for lipid biosynthetic pathways.

AIMS: It is known that Thecaphora frezii produces peanut smut that generates numerous economic losses. For this reason, it is a priority to search for control strategies. In this sense, we investigated the lipid profile of this pathogen, as possible antifungal targets, regarding polar lipid composition, fatty acid profile, and transcriptional regulation of genes involved in each stage of the development. METHOD AND RESULTS: Lipids from T. frezii teliospores, basidiospores, and hyphae were analyzed by HPLC/CAD and CG/FID. We found differences in the unsaturation levels as well as in the long-chain fatty acids along the stages. Phosphatidylcholine was the main component in the three development stages, followed by cardiolipins. Phosphatidylinositol, phosphatidylethanolamine, and lyso-phosphatidylethanolamine were found in similar amounts in all stages. Although ergosterol was not detected, we found two unsaponifiable lipids. In addition, we found transcripts that encode 28 enzymes involved in the biosynthesis of three lipids by RNA-Seq. CONCLUSIONS: Thecaphora frezii shows changes in the composition of membrane lipids in different ontogenetic stages as well as in the expression of transcripts for enzymes involved in lipid biosynthesis.

Draft Genome Sequence of Gordonia sp. Strain Campus, a Bacterium Isolated from Diesel-Contaminated Soil with Potential Use in Phytoremediation Systems.

We present the draft genome sequence of Gordonia sp. strain Campus, which was extracted from diesel-contaminated soil in Córdoba, Argentina. It was observed that this strain, in conjunction with alfalfa and poplar, has the ability to decompose diesel-contaminated soils. The data may be important for the phytoremediation of hydrocarbon-contaminated soils.

Pharmacogenomics, biomarker network, and allele frequencies in colorectal cancer.

Colorectal cancer is one of the leading causes of cancer death worldwide. Over the last decades, several studies have shown that tumor-related genomic alterations predict tumor prognosis, drug response, and toxicity. These observations have led to the development of several therapies based on individual genomic profiles. As part of these approaches, pharmacogenomics analyses genomic alterations which may predict an efficient therapeutic response. Studying these mutations as biomarkers for predicting drug response is of a great interest to improve precision medicine. We conduct a comprehensive review of the main pharmacogenomics biomarkers and genomic alterations affecting enzyme activity, transporter capacity, channels, and receptors; and therefore the new advances in CRC precision medicine to select the best therapeutic strategy in populations worldwide, with a focus on Latin America.

Rol de variantes genéticas en el desarrollo de Enfermedad Renal Crónica en pacientes con Diabetes Mellitus Tipo 2 / Role of genetic variants to the development of Chronic Kidney Disease in patients with Type 2 Diabetes Mellitus

La Enfermedad Renal Crónica (ERC) es una afección que perjudica a un gran número de pacientes. Una de las causas es la Nefropatía en pacientes con Diabetes Mellitus Tipo 2 (DM2). OBJETIVO: Indagar si las presencias de variantes genéticas contribuyen al desarrollo de ERC en pacientes con DM2. Materiales y métodos: se evaluaron criterios clínicos, bioquímicos y moleculares en 25 pacientes con DM2. Los polimorfismos se analizaron mediante PCR-RFLP para ECA (rs4646994); CDKAL1 (rs7756992); e-NOS (rs1799983) y SLC12A3 (rs11643718). RESULTADOS: El análisis estadístico mediante modelo dominante arrojaron para: ECA (rs4646994) (OR=1,33 (IC 95%) 0,25-7,01; p=0,73); CDKAL1 (rs7756992) (OR= 1 (IC 95%) 0,14-7,39; p=NA); e-NOS (rs1799983) (OR= 0,29 (IC 95%) 0,05-1,57; p=0,14) y SLC12A3 (rs11643718 (OR= 1,62 (IC 95%) 0,19-13,93; p=0,66). CONCLUSIONES: ninguna de las variantes evaluadas en los genes ECA, CDKAL1, e-NOS y SLC12A3 mostraron asociaciones positivas o negativas con el riesgo a desarrollar ERC en pacientes con DM2. (AU)

Chronic Kidney Disease (CKD) is a condition that affects a large number of patients. One cause is the Nephropathy in Type 2 Diabetes Mellitus (DM2).patients. OBJECTIVE: Evaluate if some polymorphisms contribute to CKD risk in DM2 patients. Materials and methods: clinical, biochemical and molecular parameters from 25 patients with DM2 were evaluated. The polymorphisms were analyzed by PCR-RFLP for the ECA (rs4646994); CDKAL1 (rs7756992); e-NOS (rs1799983) and SLC12A3 (rs11643718) genes. RESULTS: Statistical analysis using a dominant model showed: ACE (rs4646994) (OR = 1.33 (95% CI) 0.25-7.01, p = 0.73); CDKAL1 (rs7756992) (OR = 1 (95% CI) 0.14-7.39; p = NA); e-NOS (rs1799983) (OR = 0.29 (95% CI) 0.05- 1.57; p = 0.14) and SLC12A3 (rs11643718 (OR = 1.62 (95% CI) 0.19-) 13.93, p = 0.66). CONCLUSIONS: none of the evaluated variants in ACE, CDKAL1, e-NOS and SLC12A3 genes showed positive or negative associations with CKD risk in DM2 patients.

Dynamics of expression of two vitellogenin genes in the Chagas' disease vector Triatoma infestans: Analysis throughout pre-vitellogenesis and vitellogenesis.

The reproductive success of all oviparous species depends on vitellogenin (Vg) biosynthesis and its accumulation in the developing oocytes. The expression levels of two Vg genes (Vg1 and Vg2) were analyzed by qPCR and western blot in fat body and ovaries of adult females, at different times after ecdysis (pre-vitellogenic phase) and after blood feeding of females (vitellogenic phase). Vg genes were also evaluated in fat bodies of adult males as well as in female fifth instar nymphs. No trace of Vg mRNA was detected in adult males or in nymphs. Vg1 and Vg2 were expressed in the fat bodies and ovaries of adult females. The Vg genes start to be expressed slightly in both tissues of adult females during pre-vitellogenesis. After blood feeding, Vg1 and Vg2 were up regulated and significant levels of Vg transcripts as well as protein expression were observed in fat bodies sampled throughout vitellogenesis. During this period however, the distribution patterns of Vg1 and Vg2 transcripts showed two peaks around early and advanced vitellogenesis (days 4 and 12 post-feeding, respectively). In the ovaries, levels of mRNAs increased from the day 10 post-blood feeding onwards. In addition, the immunofluorescence assays showed a strong signal for vitellin in the yolk bodies of terminal follicles of vitellogenic females. The involvement of fat body and ovary in the synthesis of Vg suggests different roles of Vgs in supporting the growth of oocytes.

cDNA isolation and characterization of two vitellogenin genes in the Chagas' disease vector Triatoma infestans (Hemiptera, Reduviidae).

Two vitellogenin genes (Vg1 and Vg2) were identified in the Chagas' disease vector Triatoma infestans. The putative coding sequence corresponding to Vg2 was found to be 5553bp long, encoding 1851 amino acids in a single open reading frame. The comparative analysis of the deduced amino acid sequences from Vg1 and Vg2 cDNA fragments of T. infestans revealed 58.94% of identity with 76.43% of homology. The phylogenetic tree based on the complete Vg amino acid sequences of hemimetabolous insects unambiguously supported two clusters, one consisting of Vg sequences from dictyopteran and the other containing Vg sequences of hemipteran. The Vg1 and Vg2 mRNAs were detected in fat bodies and ovaries of adult females with the highest levels of both Vg transcripts in the first tissue. Quantitative PCR showed low expression of Vg2 in head and muscle of adult females, while the Vg1 transcript was not present in these organs. Neither Vg1 nor Vg2 was expressed in fifth instar nymph fat bodies or in adult male fat bodies, heads, and muscles.

Distribution of polymorphisms in cytochrome P450 2B6, histocompatibility complex P5, chemokine coreceptor 5, and interleukin 28B genes in inhabitants from the central area of Argentina.

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.

Genetic profiling of GSTP1, DPYD, FCGR2A, FCGR3A and CCND1 genes in an Argentinian population.

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.

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 (ß-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, ß-Act, and the dengue virus genome were performed. cDNA fragments for ß-Act and G6PDH were amplified for all samples, regardless of the presence or absence of viral RNA. CONCLUSIONS: Amplification of ß-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.

Clinical significance of V617F mutation of the JAK2 gene in patients with chronic myeloproliferative disorders.

OBJECTIVE: To determine the prevalence of JAK2 V617F mutation and its clinical correlation in patients with chronic myeloproliferative disorders (CMD): polycythemia vera (PV), essential thrombocythemia (ET) and idiopathic myelofibrosis (IMF). MATERIALS AND METHODS: Detection of JAK2 V617F mutation by allele specific-PCR. RESULTS: One hundred and three patients with CMD were included in the study. JAK2 V617F distribution was PV 40/45 (89%), ET 30/43 (69%), and IMF 7/15 (47%). In PV and ET patients only, 18 had thrombosis at diagnosis and 12 during follow-up (these were microvascular: 11, venous: 7 and arterial: 12); of these 28/70 (40%) were JAK2pos versus 2/18 (11%) JAK2neg; P=0.02. In a median of 4 years, two patients with PV JAK2pos evolved to myelofibrosis and one patient with PV presented in leukemic transformation (JAK2pos before and after transformation); six patients died: four patients with IMF and two patients with PV. CONCLUSIONS: We found an association between JAK2 V617F and thrombotic events in patients with PV and ET.

Estudios bioquímicos y caracterización molecular de pacientes portadores de ß talasemia en Córdoba, Argentina / Biochemical studies and molecular characterization of ß-thalassemia carriers in Córdoba, Argentina

Las ß talasemias constituyen un grupo heterogéneo de anemias microcíticas, causadas por una producción reducida (ß+) o ausencia (ߺ) de síntesis de cadenas de globina ß. La diferente expresividad clínica de estas patologías resulta de la combinación de ambas posibilidades o de cada una de ellas con el gen normal. Dado que este país ha recibido distintas corrientes migratorias, y que

Estudios bioquímicos y caracterización molecular de pacientes portadores de ß talasemia en Córdoba, Argentina / Biochemical studies and molecular characterization of ß-thalassemia carriers in Córdoba, Argentina

Las ß talasemias constituyen un grupo heterogéneo de anemias microcíticas, causadas por una producción reducida (ß+) o ausencia (ߺ) de síntesis de cadenas de globina ß. La diferente expresividad clínica de estas patologías resulta de la combinación de ambas posibilidades o de cada una de ellas con el gen normal. Dado que este país ha recibido distintas corrientes migratorias, y que no son pocos los portadores ß talasémicos, se intentó identificar por primera vez qué tipo de mutaciones son las más frecuentes en este medio. Para realizarlo, se contó con doce pacientes provenientes de la ciudad de Córdoba que fueron evaluados bioquímicamente con: hematocrito, hemoglobina, porcentaje de reticulocitos, electroforesis de hemoglobinas a pH alcalino, hierro sérico, transferrina, hemoglobina fetal y morfología de los glóbulos rojos. La obtención de un volumen corpuscular medio inferior a 75 fl como así también cantidades superiores de Hb A2 a 2 por ciento fueron los criterios más importantes para la selección de los pacientes. El uso de PCR con uno de los oligonucleótidos de cada set que posee un segmento adicional rico en GC y una posterior electroforesis en un gel desnaturalizante (DGGE), permitió efectuar la identificación de mutaciones puntuales en el gen de la globulina ß. De esta manera se obtuvieron los siguientes resultados: 41,7 por ciento ߺ talasemia (codón 39 C -> T), 50,0 por ciento ß+ talasemia (IVS1 nucleótico 110 G -> A) y 8,3 por ciento ߺ talasemia (IVS1 nucleótico 1 G ->A)