Validation of a multiplex reverse transcription and pre-amplification method using TaqMan(®) MicroRNA assays.

Since the discovery of microRNAs (miRNAs), different approaches have been developed to label, amplify and quantify miRNAs. The TaqMan(®) technology, provided by Applied Biosystems (ABIs), uses a stem-loop reverse transcription primer system to reverse transcribe the RNA and amplify the cDNA. This method is widely used to identify global differences between the expression of 100s of miRNAs across comparative samples. This technique also allows the quantification of the expression of targeted miRNAs to validate observations determined by whole-genome screening or to analyze few specific miRNAs on a large number of samples. Here, we describe the validation of a method published by ABIs on their web site allowing to reverse transcribe and pre-amplify multiple miRNAs and snoRNAs simultaneously. The validation of this protocol was performed on human muscle and plasma samples. Fast and cost efficient, this method achieves an easy and convenient way to screen a relatively large number of miRNAs in parallel

Altered expression of β-galactosidase-1-like protein 3 (Glb1l3) in the retinal pigment epithelium (RPE)-specific 65-kDa protein knock-out mouse model of Leber’s congenital amaurosis

Purpose: In this study, we investigated the expression of the gene encoding beta-galactosidase (Glb)-1-like protein 3 (Glb1l3), a member of the glycosyl hydrolase 35 family, during retinal degeneration in the retinal pigment epithelium (RPE)-specific 65-kDa protein knockout (Rpe65(-/-)) mouse model of Leber congenital amaurosis (LCA). Additionally, we assessed the expression of the other members of this protein family, including beta-galactosidase-1 (Glb1), beta-galactosidase-1-like (Glb1l), and beta-galactosidase-1-like protein 2 (Glb1l2).Methods: The structural features of Glb1l3 were assessed using bioinformatic tools. mRNA expression of Glb-related genes was investigated by oligonucleotide microarray, real-time PCR, and reverse transcription (RT) -PCR. The localized expression of Glb1l3 was assessed by combined in situ hybridization and immunohistochemistry.Results: Glb1l3 was the only Glb-related member strongly downregulated in Rpe65(-/-) retinas before the onset and during progression of the disease. Glb1l3 mRNA was only expressed in the retinal layers and the RPE/choroid. The other Glb-related genes were ubiquitously expressed in different ocular tissues, including the cornea and lens. In the healthy retina, expression of Glb1l3 was strongly induced during postnatal retinal development; age-related increased expression persisted during adulthood and aging.Conclusions: These data highlight early-onset downregulation of Glb1l3 in Rpe65-related disease. They further indicate that impaired expression of Glb1l3 is mostly due to the absence of the chromophore 11-cis retinal, suggesting that Rpe65 deficiency may have many metabolic consequences in the underlying neuroretina

MONITORIAMENTO DE SALMONELOSE EM AVES DE PRODUÇÃO, ULTILIZANDO O MÉTODO DE SOROAGLUTINAÇÃO RÁPIDA EM PLACA

The frequency of was evaluated Salmonella Pullorum and Salmonella Gallinarum in 150 laying hens belonging to Hy Line (110 birds) and Dekalb brown (40 birds) and 150 broilers (Cobb line) housed in the poultry sector of the Department of Animal Science, Federal University of Paraíba - CCA - Areia - PB. As a serological screening method, the rapid serum agglutination test (SAR) was used. Of the 150 serological samples from laying hens none of them presented positive serology to the test. Of the 150 serological samples collected from broilers only 2 were seropositive, resulting in a frequency of 1.33% relative to broilers and 0.66% of frequency in relation to the total of animals analyzed.Avaliou-se a frequência de Salmonella Pullorum e Salmonella Gallinarum em 150 galinhas poedeiras pertencentes as linhagens Hy Line (110 aves) e Dekalb brown (40 aves) e de 150 frangos de corte (linhagem Cobb) alojadas no setor de Avicultura do Departamento de Zootecnia, da Universidade Federal da Paraíba – CCA – Areia – PB. Como método de triagem sorológica foi utilizado o teste de Soroaglutinação Rápida (SAR). Das 150 amostras sorológicas provenientes de galinhas poedeiras nenhuma apresentou sorologia positiva ao teste. De 150 amostras sorológicas colhidas de frangos de corte apenas 2 foram soropositivas resultando em umas frequência de 1,33% relativo aos frangos e 0,66% de frequência em relação ao total de animais analisados

Évreux – Zac Joséphine, rue Joséphine

Identifiant de l'opération archéologique : 7602 Date de l'opération : 1999 (SP) Inventeur(s) : Le Cain Bérengère (AFAN) Le terrain est placé entre la rue Joséphine et le cours canalisé de l'Iton. Il se trouve à l'extérieur de la ville du Haut-Empire. Au Moyen Âge, le site, placé à l'ouest du centre urbain, était en dehors du castrum et de l'extension maximale connue de l'enceinte du bourg (paroisse Saint-Pierre), mais proche de l'abbaye Saint-Taurin, voir site K sur le plan général de la vill..

“Montrer ce qui est vivant”

On se rappelle la perméabilité avec laquelle Xavier Le Roy envisageait la « production » – dispositif de production autant que production d’un produit – dans Self Interview : « X : C’est pourquoi il me semble nécessaire d’inclure le maximum de facteurs du contexte dans lequel nous travaillons comme questions inhérentes au travail pour ne pas se trouver dans un système d’exclusion ou de séparation entre le sujet de travail ou d’expérimentation, son contexte et sa représentation. MERCI ISABELLE..

Pelagic population dynamics of Aurelia sp in French Mediterranean lagoons

The pelagic dynamics of the cosmopolitan scyphozoan Aurelia sp. was investigated in three French Mediterranean lagoons, Thau, Berre and Bages-Sigean, which harbour resident populations. The annual cycles showed a common univoltine pattern in all lagoons where the presence of pelagic stages in the water column lasted similar to 8 months. Field observations showed a release of ephyrae in winter time followed by pronounced growth between April and July, when individuals reached the largest sizes, before disappearing from the water column. Maximum abundance of ephyrae and medusae were registered in Thau. Medusae abundance attained a maximum of 331 ind 100 m(-3) in Thau, 18 ind 100 m(-3) in Berre and 7 ind 100 m(-3) in Bages-Sigean lagoons. Temperature and zooplankton abundance appeared as leading factors of growth, where Bages-Sigean showed the population with higher growth rates (2.66 mm day(-1)) and maximum size (32 cm), followed by Thau (0.57-2.56 mm day(-1); 22.4 cm) and Berre (1.57-2.22 mm day(-1); 17 cm). The quantification of environmental windows used by the species showed wider ranges than previously reported in the Mediterranean Sea, which suggests a wide ecological plasticity of Aurelia spp. populations in north-western Mediterranean lagoons.GELAMED project; Total foundation [189 - "Recherche" 18902 C]; EC2CO "Ecosphere Continentale et Cotiere" programme through the DYNAMO project; OSU-OREM

Bronchial responses to substance P after antigen challenge in the guinea-pig: in vivo and in vitro studies

The effect of antigen challenge on the airway responses to substance P and on the epithelial neutral endopeptidase (NEP) activity was investigated in aerosol sensitized guinea-pigs. In vivo, bronchial responses to aerosolized substance P were similar to the responses observed in antigen-challenged guinea-pigs and in the control groups. In contrast, when the guinea-pigs were pretreated with the NEP inhibitor, phosphoramidon, a significant increase in the airway responses to substance P was observed after antigen challenge in vivo. However, in vitro, the contractile responses of the tracheal smooth muscle to substance P were similar between groups of guinea-pigs, in respect to the presence or absence of the epithelium and/or phosphoramidon. Histological studies showed an accumulation of eosinophils in the tracheal submucosa after antigen challenge and intact epithelial cells. These results show that in vivo bronchial hyperresponsiveness to substance P after antigen challenge in the guinea-pig is not associated with increased responses of the smooth muscle to exogenous SP in vitro. In addition, the results with phosphoramidon suggest that loss of NEP activity cannot account for the in vivo bronchial hyperresponsiveness to substance P presently observed

Emergency surgery for splenic flexure cancer: results of the SFC Study Group database

Colectomía; Mortalidad; Carcinoma del ángulo esplénicoColectomia; Mortalitat; Carcinoma de l'angle esplènicColectomy; Mortality; Splenic flexure carcinomaBackground The effectiveness of surgical treatment for splenic flexure carcinomas (SFCs) in emergency settings remains unexplored. This study aims to compare the perioperative and long-term outcomes of different alternatives for emergency SFC resection. Method This multicenter retrospective study was based on the SFC Study Group database. For the present analysis, SFC patients were selected if they had received emergency surgical resection with curative intent between 2000 and 2018. Extended right colectomy (ERC), left colectomy (LC), and segmental left colectomy (SLC) were evaluated and compared. Results The study sample was composed of 90 SFC patients who underwent emergency ERC (n = 55, 61.1%), LC (n = 18, 20%), or SLC (n = 17, 18.9%). Bowel obstruction was the most frequent indication for surgery (n = 75, 83.3%), and an open approach was chosen in 81.1% of the patients. A higher incidence of postoperative complications was observed in the ERC group (70.9%) than in the LC (44.4%) and SLC groups (47.1%), with a significant procedure-related difference for severe postoperative complications (Dindo-Clavien ≥ III; adjusted odds ratio for ERC vs. LC:7.23; 95% CI 1.51-34.66; p = 0.013). Anastomotic leakage occurred in 8 (11.2%) patients, with no differences between the groups (p = 0.902). R0 resection was achieved in 98.9% of the procedures, and ≥ 12 lymph nodes were retrieved in 92.2% of patients. Overall and disease-free survival rates at 5 years were similar between the groups and were significantly associated with stage pT4 and the presence of synchronous metastases. Conclusion In the emergency setting, ERC and open surgery are the most frequently performed procedures. ERC is associated with increased odds of severe postoperative complications when compared to more conservative SFC resections. Nonetheless, all the alternatives seem to provide similar pathologic and long-term outcomes, supporting the oncological safety of more conservative resections for emergency SFCs.No authors received funding or resources in relation to this article

Beyond aspect: will be -ing and shall be -ing

This article discusses the synchronic status and diachronic development of will be -ing and shall be -ing (as in I’ll be leaving at noon).2 Although available since at least Middle English, the constructions did not establish a significant foothold in standard English until the twentieth century. Both types are also more prevalent in British English (BrE) than American English (AmE). We argue that in present-day usage will/shall be -ing are aspectually underspecified: instances that clearly construe a situation as future-in-progress are in the minority. Similarly, although volition-neutrality has been identified as a key feature of will/shall be -ing, it is important to take account of other, generally richer meanings and associations, notably ‘future-as-matter-of-course’ (Leech 2004), ‘already-decided future’ (Huddleston & Pullum et al. 2002) and non-agentivity. Like volition-neutrality, these characteristics appear to be relevant not only in contemporary use, but also in their historical expansion. We show that the construction has evolved from progressive aspect towards more subjectivised evidential meaning

Intense exercise training induces adaptation in expression and responsiveness of cardiac β-adrenoceptors in diabetic rats

<p>Abstract</p> <p>Background</p> <p>Informations about the effects of intense exercise training on diabetes-induced myocardial dysfunctions are lacking. We have examined the effects of intense exercise training on the cardiac function of diabetic rats, especially focusing on the Langendorff β-adrenergic responsiveness and on the β-adrenoceptors protein expression.</p> <p>Methods</p> <p>Control or Streptozotocin induced-diabetic male Wistar rats were randomly assigned to sedentary or trained groups. The training program consisted of 8 weeks running on a treadmill (10° incline, up to 25 m/min, 60 min/day) and was considered to be intense for diabetic rats.</p> <p>Results</p> <p>This intense exercise training amplified the <it>in vivo </it>diabetes-induced bradycardia. It had no effect on Langendorff basal cardiac contraction and relaxation performances in control and diabetic rats. In diabetic rats, it accentuated the Langendorff reduced responsiveness to β-adrenergic stimulation. It did not blunt the diabetes-induced decrease of β1-adrenoceptors protein expression, displayed a significant decrease in the β2-adrenoceptors protein expression and normalized the β3-adrenoceptors protein expression.</p> <p>Conclusions</p> <p>Intense exercise training accentuated the decrease in the myocardial responsiveness to β-adrenergic stimulation induced by diabetes. This defect stems principally from the β2-adrenoceptors protein expression reduction. Thus, these results demonstrate that intense exercise training induces specific effects on the β-adrenergic system in diabetes.</p