Effects of Dicer and Argonaute down-regulation on mRNA levels in human HEK293 cells

RNA interference and the microRNA (miRNA) pathway can induce sequence-specific mRNA degradation and/or translational repression. The human genome encodes hundreds of miRNAs that can post-transcriptionally repress thousands of genes. Using reporter constructs, we observed that degradation of mRNAs bearing sites imperfectly complementary to the endogenous let-7 miRNA is considerably stronger in human HEK293 than HeLa cells. The degradation did not result from the Ago2-mediated endonucleolytic cleavage but it was Dicer- and Ago2-dependent. We used this feature of HEK293 to address the size of a pool of transcripts regulated by RNA silencing in a single cell type. We generated HEK293 cell lines depleted of Dicer or individual Ago proteins. The cell lines were used for microarray analyses to obtain a comprehensive picture of RNA silencing. The 3'-untranslated region sequences of a few hundred transcripts that were commonly up-regulated upon Ago2 and Dicer knock-downs showed a significant enrichment of putative miRNA-binding sites. The up-regulation upon Ago2 and Dicer knock-downs was moderate and we found no evidence, at the mRNA level, for activation of silenced genes. Taken together, our data suggest that, independent of the effect on translation, miRNAs affect levels of a few hundred mRNAs in HEK293 cells

Post-Retained Single Crowns versus Fixed Dental Prostheses: A 7-Year Prospective Clinical Study

Biomechanical integrity of endodontically treated teeth (ETT) is often compromised. Degree of hard tissue loss and type of final prosthetic restoration should be carefully considered when making a treatment plan. The objective of this prospective clinical trial was to assess the influence of the type of prosthetic restoration as well as the degree of hard tissue loss on 7-y clinical performance of ETT restored with fiber posts. Two groups (n = 60) were defined depending on the type of prosthetic restoration needed: 1) single unit porcelain-fused-to-metal (PFM) crowns (SCs) and 2) 3- to 4-unit PFM fixed dental prostheses (FDPs), with 1 healthy and 1 endodontically treated and fiber post-restored abutment. Within each group, samples were divided into 2 subgroups (n = 30) according to the amount of residual coronal tissues after abutment buildup and final preparation: A) >50% of coronal residual structure or B) equal to or <50% of coronal residual structure. The clinical outcome was assessed based on clinical and intraoral radiographic examinations at the recalls after 6, 12, 24, 36, 48, and 84 mo. Data were analyzed by Kaplan-Meier log-rank test and Cox regression analysis (P < 0.05). The overall 7-y survival rate of ETT restored with fiber post and either SCs or FDPs was 69.2%. The highest 84-mo survival rate was recorded in group 1A (90%), whereas teeth in group 2B exhibited the lowest performance (56.7% survival rate). The log-rank test detected statistically significant differences in survival rates among the groups (P = 0.048). Cox regression analysis revealed that the amount of residual coronal structure (P = 0.041; hazard ratio [HR], 2.026; 95% confidence interval [CI] for HR, 1.031–3.982) and the interaction between the type of prosthetic restoration and the amount of residual coronal structure (P = 0.024; HR, 1.372; 95% CI for HR, 1.042–1.806) were statistically significant factors for survival (ClinicalTrials.gov NCT01532947)

Mechanical detection and pain thresholds: comparability of devices using stepped and ramped stimuli

Quantitative sensory testing is used to assess somatosensory function in humans. The protocol of the German Research Network on Neuropathic Pain (DFNS) provides comprehensive normative values using defined tools; however, some of these may not be feasible in low-resource settings. Objectives: To compare the standard DFNS devices for assessment of mechanosensory function to a low resource tool, the Sorri-Bauru-monofilaments. Methods: Mechanical detection thresholds (MDT), pain thresholds (MPT), and suprathreshold pinprick ratings (pain sensitivity: MPS) were measured over cheek, hand dorsum, and fingertip in 13 healthy subjects (7 female, aged 21-44 years). Mechanical detection threshold was assessed with DFNS standard glass monofilaments (0.25-512 mN, 0.5 mm tip) and nylon monofilaments (Sorri-Bauru; 0.5-3000 mN). MPT was assessed with DFNS standard cylindrical probes (8-512 mN, 0.25 mm tip), Sorri-Bauru monofilaments, and with ramped stimuli using an electronic von Frey aesthesiometer (10 mN/s or 100 mN/s, 0.20 mm tip). MPS was measured in response to stepped and ramped pinpricks (128 and 256 mN). Results: Mechanical detection thresholds were the same for DFNS and Sorri-Bauru monofilaments. For MPT, Sorri-Bauru filaments yielded lower values than PinPricks over face but not hand. Pain thresholds were higher at all test sites for ramped than stepped pinpricks (P < 0.01). Suprathreshold ratings were lower for ramped than stepped pinpricks (P < 0.05). Conclusion: Sorri-Bauru filaments are acceptable substitutes for DFNS standards in estimating tactile sensitivity, but are not consistent with standard probes for pinprick sensitivity because of their nonstandardized tips. Ramped stimuli overestimated MPT and underestimated MPS due to reaction time artefacts and therefore need their own normative values

The changing patterns of group politics in Britain

Two interpretations of ways in which group politics in Britain have presented challenges to democracy are reviewed: neo-corporatism or pluralistic stagnation and the rise of single issue interest groups. The disappearance of the first paradigm created a political space for the second to emerge. A three-phase model of group activity is developed: a phase centred around production interests, followed by the development of broadly based 'other regarding' groups, succeeded by fragmented, inner directed groups focusing on particular interests. Explanations of the decay of corporatism are reviewed. Single issue group activity has increased as party membership has declined and is facilitated by changes in traditional media and the development of the internet. Such groups can overload the policy-making process and frustrate depoliticisation. Debates about the constitution and governance have largely ignored these issues and there is need for a debate

Composición y estructura de la ictiofauna del río Hondo, México-Belice, con base en el uso del arpón

ResumenSe determinó la composición y estructura de la ictiofauna del río Hondo y la variación espacial de éstas en función de los parámetros ambientales. Se realizaron 3 muestreos durante marzo, abril y mayo de 2012, en 6 sitios a lo largo del río. Los datos usados para determinar la estructura se obtuvieron con ayuda de un arpón de 58.5cm, utilizado durante 2h de buceo libre en cada punto de muestreo. Para complementar la lista de especies se utilizaron redes agalleras, palangres, nasas y anzuelos. Además, se realizó una revisión bibliográfica y se consultó material de la colección de peces de ECOSUR en Chetumal. La lista sistemática se conformó por 40 especies en 33 géneros, 18 familias y 11 órdenes. La familia mejor representada fue Cichlidae, incluida la especie exótica Oreochromis niloticus. Los parámetros ambientales que mejor describieron la variación de la distribución y abundancia en los peces en el río Hondo fueron: la anchura del río, distancia a la boca, transparencia, profundidad, concentración de oxígeno disuelto y conductividad. La relación entre las especies y las variables ambientales fue alta en ambos ejes (0.89 y 0.79). Ambientalmente, el río se puede zonificar en 3 partes, aunque su ictiofauna puede dividirse en 2 conjuntos principales.AbstractComposition and structure of the Hondo River ichthyofauna and its spatial variation were determined as a function of environmental parameters. Six sites along the river were sampled in March, April and May 2012. Data for estimating structure of fish assemblages was obtained by means of a 58.5cm harpoon during 2 hours of free diving at each sampling site. The species list for the study area was completed with the aid of gillnets, longlines, double-cone traps, and hook-and-line. In addition, a bibliographical research performed as well as the ichthyological collection of ECOSUR at Chetumal was checked. The systematic list includes 40 species in 33 genera, 18 families and 11 orders. The richest family was Cichlidae, including the exotic Oreochromis niloticus. The environmental parameters that best described distribution and abundance of fish in the Hondo River were river width, distance to mouth, transparency, depth, dissolved oxygen concentration, and conductivity. Correlation between species and environmental parameters was high in both axis (0.89 and 0.79). The river was classified into 3 environmental zones, although its fish fauna could be divided in 2 main assemblages

Analysis of S. pombe SIN protein association to the SPB reveals two genetically separable states of the SIN.

The Schizosaccharomyces pombe septation initiation network (SIN) regulates cytokinesis, and asymmetric association of SIN proteins with the mitotic spindle pole bodies (SPBs) is important for its regulation. Here, we have used semi-automated image analysis to study SIN proteins in large numbers of wild-type and mutant cells. Our principal conclusions are: first, that the association of Cdc7p with the SPBs in early mitosis is frequently asymmetric, with a bias in favour of the new SPB; second, that the early association of Cdc7p-GFP to the SPB depends on Plo1p but not Spg1p, and is unaffected by mutations that influence its asymmetry in anaphase; third, that Cdc7p asymmetry in anaphase B is delayed by Pom1p and by activation of the spindle assembly checkpoint, and is promoted by Rad24p; and fourth, that the length of the spindle, expressed as a fraction of the length of the cell, at which Cdc7p becomes asymmetric is similar in cells dividing at different sizes. These data reveal that multiple regulatory mechanisms control the SIN in mitosis and lead us to propose a two-state model to describe the SIN

Constraints of small-scale irrigated fodder production and nutrition assessment for livestock feed, a case study in Ethiopia

Livestock is an integral part of the agricultural system in sub-Saharan Africa, serving as a food source, income, fertilizer, and power for farming and transportation. However, the productivity of the livestock system has been hampered due to a lack of sufficient quantity and quality feed. This study evaluates the gaps and constraints of fodder and nutritional potential for livestock feed using small-scale irrigation (SSI). The study comprised of 30 randomly selected farmers from two different ecological zones in Ethiopia. Half of the farmers cultivated Napier grass (Pennisetum purpureum) in the Robit watershed in northern Ethiopia, and the other half cultivated mixed vetch (Lathyrus cicera) and oats (Avena sativa) in Lemo watershed in southern Ethiopia. The Soil and Water Assessment Tool (SWAT) and Agricultural Policy Environmental eXtender (APEX) were applied in an integrated manner to assess the impacts of SSI at the watershed and field-scale levels, respectively. The watershed-scale analysis showed that there is a substantial amount of surface runoff and shallow groundwater recharge that could be used for dry season fodder production using irrigation. Field data calibrated APEX model indicated that Napier yield could be maximized with 550 mm of water in Robit watershed. While in the Lemo watershed, maximum vetch and oats yield may be achieved with 250 mm of water. The major constraints for Napier and oats production in the study sites were soil fertility, especially nitrogen and phosphorus, and vetch production was limited by high temperature. Fodder samples were collected at the time of harvest to evaluate feed quality. The nutritional analysis indicated that Napier grass has a higher dry matter and ash (mineral) content compared to oats and vetch. However, vetch has higher crude protein content (18%) compared to Napier (10%) and oats (6%). Overall the study indicated that cultivating vetch provided superior performance in terms of providing quality feed and environmental services

Are language production problems apparent in adults who no longer meet diagnostic criteria for attention-deficit/hyperactivity disorder?

In this study, we examined sentence production in a sample of adults (N = 21) who had had attention-deficit/hyperactivity disorder (ADHD) as children, but as adults no longer met DSM-IV diagnostic criteria (APA, 2000). This “remitted” group was assessed on a sentence production task. On each trial, participants saw two objects and a verb. Their task was to construct a sentence using the objects as arguments of the verb. Results showed more ungrammatical and disfluent utterances with one particular type of verb (i.e., participle). In a second set of analyses, we compared the remitted group to both control participants and a “persistent” group, who had ADHD as children and as adults. Results showed that remitters were more likely to produce ungrammatical utterances and to make repair disfluencies compared to controls, and they patterned more similarly to ADHD participants. Conclusions focus on language output in remitted ADHD, and the role of executive functions in language production

Together forever? Explaining exclusivity in party-firm relations

Parties and firms are the key actors of representative democracy and capitalism respectively and the dynamic of attachment between them is a central feature of any political economy. This is the first article to systematically analyse the exclusivity of party-firm relations. We consider exclusivity at a point in time and exclusivity over time. Does a firm have a relationship with only one party at a given point in time, or is it close to more than one party? Does a firm maintain a relationship with only one party over time, or does it switch between parties? Most important, how do patterns of exclusivity impact on a firm’s ability to lobby successfully? We propose a general theory, which explains patterns of party-firm relations by reference to the division of institutions and the type of party competition in a political system. A preliminary test of our theory with Polish survey data confirms our predictions, establishing a promising hypothesis for future research