Identification of proteaginous pea cultivars (Pisum sativum L.) using microsatellites molecular markers

Abstract: Conventionally morphological descriptors are routinely used for establishing the identity of varieties. This kind of descriptors has some disadvantages, namely most of them are quantitative, controlled by several pairs of genes, and their expression is influenced by environmental factors. Molecular markers have a potential to facilitate this procedure, increase the reliability of decisions, and substantially save the time and space needed for experiments. In this study we intended to identify 20 cultivars of proteaginous pea (Pisum sativum L.), registered in the Community Catalog of Varieties, by microsatellites molecular markers. After DNA extraction, seven different loci were analyzed. PCR amplifications were conducted and the resulting fragments were separated on an 3,5% MS-8 agarose gel in TBE buffer, at 90V/h. The gels were analyzed for the presence/ absence of bands and a table with binary code was made. The data were processed with the statistical software NTSYS-pc, using the SIMQUAL module and Jaccard similarity coefficient, followed by UPGMA cluster analysis. With the analysis of six polymorphic loci was possible to distinguish almost all of cultivars. The most informative loci were AD61 and AB53. The cluster analysis of SSR markers separated the pea genotypes into two distinct clusters. The first cluster included the five cultivars: Isard, Cartouche, Audit, Corrent and James. The second cluster included the remaining fifteen cultivars and was further divided in two subclusters. The first subcluster had the Portuguese genotype Grisel and second subcluster contained the remaining fourteen cultivars. In this subcluster Ideal and Alezan had 100 percent similarity. There was a low number of heterozygous loci which is consistent with the nature of self pollinated species. The results showed a high potential and resolving power of SSR markers in distinctness assessment. SSR markers might also be useful in Pisum sativum L. germplasm management and genetic diversity studies.The present study was financially by the EU within the POCTEP – “Programa Operacional de Cooperação Transfronteiriça Espanha - Portugal”, Project 0186_AGROCELE_3_E

Laboratório de Biologia.

Laboratório de Biologia.info:eu-repo/semantics/publishedVersio

Identificação de cultivares por microssatélites - aplicação à ervilha proteaginosa

Comunicação apresentada no Seminário "Cereais e Leguminosas: Novas Perspectivas para a Beira Interior", que se realizou em 29 de Junho, em Castelo Branco, na Escola Superior Agrária do Instituto Politécnico de Castelo Branco.O objectivo deste trabalho foi realizar a identificação, por marcadores moleculares do tipo microssatélites, de 20 cultivares de ervilha proteaginosa, inscritas no catálogo comunitário de variedades. A extracção de DNA foi realizada com o DNAeasy Plant Mini Kit. Foram estudados 7 marcadores moleculares SSRs com o objectivo de detectar polimorfismos. Os fragmentos resultantes de amplificação foram separados por electroforese em gel de agarose MS-8, a 3,5% (w/v), em tampão TBE 1x, com coloração com brometo de etídio. Foi usado o marcador 100pb ladder da Biotools. Para cada locus foi calculado o valor PIC (Polymorphic Index Content) e os dados foram processados com utilização do software estatístico NTSYS-pc, vs. 2.21k. Foi utilizado o módulo SIMQUAL com coeficiente de similaridade de Jaccard, seguido de análise de cluster UPGMA (unweighted pair-group method with arithmetic mean). No total dos 7 loci foram registados 20 alelos. Foram detectados polimorfismos em 6 dos loci estudados, tendo sido construída uma tabela de código binário com os resultados obtidos. O valor PIC variou entre 0,42 (A9) e 0,58 (AB53). Registou-se a existência de reduzido número de heterozigóticos o que é consentâneo com o facto de ser uma espécie autogâmica. Com base em 6 marcadores SSRs é possível distinguir praticamente todas as cultivares. Apenas as cultivares Ideal e Alezan apresentam perfis idênticos para os 6 loci utilizados na análise identificativa. O dendrograma representante das relações genéticas existentes entre cultivares, construído por análise de cluster UPGMA, com base nos coeficientes de similaridade de Jaccard, permite constatar a existência de dois grandes grupos. No primeiro grupo incluem-se as cultivares Cleopatra, Alhambra, Ideal, Alezan, Pixel, Lumina, Cherokee, Gregor, Livia, Guifilo, Guifredo, Onix, Arthur, Enduro e Grisel. Esta última, de origem portuguesa, aparece geneticamente distanciada das restantes. No segundo grupo incluem-se as cultivares Isard, Cartouche, Audi, Corrent e James

Prognostic value of Dna and Mrna E6/e7 of human papillomavirus in the evolution of cervical intraepithelial neoplasia grade 2

Objective: This study aimed at evaluating whether human papillomavirus (HPV) groups and E6/E7 mRNA of HPV 16, 18, 31, 33, and 45 are prognostic of cervical intraepithelial neoplasia (CIN) 2 outcome in women with a cervical smear showing a low-grade squamous intraepithelial lesion (LSIL).Methods: This cohort study included women with biopsy-confirmed CIN 2 who were followed up for 12 months, with cervical smear and colposcopy performed every three months.Results: Women with a negative or low-risk HPV status showed 100% CIN 2 regression. The CIN 2 regression rates at the 12-month follow-up were 69.4% for women with alpha-9 HPV versus 91.7% for other HPV species or HPV-negative status (P < 0.05). For women with HPV 16, the CIN 2 regression rate at the 12-month follow-up was 61.4% versus 89.5% for other HPV types or HPV-negative status (P < 0.05). The CIN 2 regression rate was 68.3% for women who tested positive for HPV E6/E7 mRNA versus 82.0% for the negative results, but this difference was not statistically significant.Conclusions: The expectant management for women with biopsy-confirmed CIN 2 and previous cytological tests showing LSIL exhibited a very high rate of spontaneous regression. HPV 16 is associated with a higher CIN 2 progression rate than other HPV infections. HPV E6/E7 mRNA is not a prognostic marker of the CIN 2 clinical outcome, although this analysis cannot be considered conclusive. Given the small sample size, this study could be considered a pilot for future larger studies on the role of predictive markers of CIN 2 evolution. © the authors.This study aimed at evaluating whether human papillomavirus (HPV) groups and E6/E7 mRNA of HPV 16, 18, 31, 33, and 45 are prognostic of cervical intraepithelial neoplasia (CIN) 2 outcome in women with a cervical smear showing a low-grade squamous intraepithelial lesion (LSIL).Methods: This cohort study included women with biopsy-confirmed CIN 2 who were followed up for 12 months, with cervical smear and colposcopy performed every three months.Results: Women with a negative or low-risk HPV status showed 100% CIN 2 regression. The CIN 2 regression rates at the 12-month follow-up were 69.4% for women with alpha-9 HPV versus 91.7% for other HPV species or HPV-negative status (P < 0.05). For women with HPV 16, the CIN 2 regression rate at the 12-month follow-up was 61.4% versus 89.5% for other HPV types or HPV-negative status (P < 0.05). The CIN 2 regression rate was 68.3% for women who tested positive for HPV E6/E7 mRNA versus 82.0% for the negative results, but this difference was not statistically significant.Conclusions: The expectant management for women with biopsy-confirmed CIN 2 and previous cytological tests showing LSIL exhibited a very high rate of spontaneous regression. HPV 16 is associated with a higher CIN 2 progression rate than other HPV infections. HPV E6/E7 mRNA is not a prognostic marker of the CIN 2 clinical outcome, although this analysis cannot be considered conclusive. Given the small sample size, this study could be considered a pilot for future larger studies on the role of predictive markers of CIN 2 evolution9152

Reduction of blood nitric oxide levels is associated with clinical improvement of the chronic pelvic pain related to endometriosis

The objective of this prospective study was to determine the plasma levels of nitric oxide (NO) in women with chronic pelvic pain secondary to endometriosis (n=24) and abdominal myofascial pain syndrome (n=16). NO levels were measured in plasma collected before and 1 month after treatment. Pretreatment NO levels (μM) were lower in healthy volunteers (47.0±12.7) than in women with myofascial pain (64.2±5.0, P=0.01) or endometriosis (99.5±12.9, P<0.0001). After treatment, plasma NO levels were reduced only in the endometriosis group (99.5±12.9 vs 61.6±5.9, P=0.002). A correlation between reduction of pain intensity and reduction of NO level was observed in the endometriosis group [correlation = 0.67 (95%CI = 0.35 to 0.85), P<0.0001]. Reduction of NO levels was associated with an increase of pain threshold in this group [correlation = -0.53 (-0.78 to -0.14), P<0.0001]. NO levels appeared elevated in women with chronic pelvic pain diagnosed as secondary to endometriosis, and were directly associated with reduction in pain intensity and increase in pain threshold after treatment. Further studies are needed to investigate the role of NO in the pathophysiology of pain in women with endometriosis and its eventual association with central sensitization.484363369CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQSem informaçã

The impact of time shifting on TV consumption and ad viewership

In this paper we study the impact of time shifting on TV consumption and ad viewership. We analyze the results of a field experiment in which a random sample of “triple-play” households were given a set of premium TV channels broadcasting popular movies and TV shows without commercial breaks. A random subset of these households were given access to these channels with time shifting (automated cloud recording for later viewing or rewinding of broadcasted programs), while the remainder were not. This design allowed us to identify the effects of time shifting on TV consumption. On average, we found that receiving access to the channels with time shifting increased total TV consumption because it increased time-shifted viewership while leaving live viewership unchanged. The increase in the live viewership of these channels was similar to the reduction in the live viewership of the originally available channels, resulting in a net zero effect on live viewership. It appears that time shifting does not change the concentration of live viewership, but it does increase the concentration of total TV viewership, because it is used disproportionately to watch the most popular programs. Finally, we found that time shifting does not change the likelihood of skipping ads during live viewership, suggesting that households