The Roles of Individual Mammalian Argonautes in RNA Interference In Vivo

Argonaute 2 (Ago2) is the only mammalian Ago protein capable of mRNA cleavage. It has been reported that the activity of the short interfering RNA targeting coding sequence (CDS), but not 3′ untranslated region (3′UTR) of an mRNA, is solely dependent on Ago2 in vitro. These studies utilized extremely high doses of siRNAs and overexpressed Ago proteins, as well as were directed at various highly expressed reporter transgenes. Here we report the effect of Ago2 in vivo on targeted knockdown of several endogenous genes by siRNAs, targeting both CDS and 3′UTR. We show that siRNAs targeting CDS lose their activity in the absence of Ago2, whereas both Ago1 and Ago3 proteins contribute to residual 3′UTR-targeted siRNA-mediated knockdown observed in the absence of Ago2 in mouse liver. Our results provide mechanistic insight into two components mediating RNAi under physiological conditions: mRNA cleavage dependent and independent. In addition our results contribute a novel consideration for designing most efficacious siRNA molecules with the preference given to 3′UTR targeting as to harness the activity of several Ago proteins.Alnylam Pharmaceuticals (Firm

Induced pseudoscalar coupling of the proton weak interaction

The induced pseudoscalar coupling $g_p$ is the least well known of the weak coupling constants of the proton's charged--current interaction. Its size is dictated by chiral symmetry arguments, and its measurement represents an important test of quantum chromodynamics at low energies. During the past decade a large body of new data relevant to the coupling $g_p$ has been accumulated. This data includes measurements of radiative and non radiative muon capture on targets ranging from hydrogen and few--nucleon systems to complex nuclei. Herein the authors review the theoretical underpinnings of $g_p$, the experimental studies of $g_p$, and the procedures and uncertainties in extracting the coupling from data. Current puzzles are highlighted and future opportunities are discussed.Comment: 58 pages, Latex, Revtex4, prepared for Reviews of Modern Physic

Precision Determination of the Neutron Spin Structure Function g1n

We report on a precision measurement of the neutron spin structure function $g^n_1$ using deep inelastic scattering of polarized electrons by polarized ^3He. For the kinematic range 0.014<x<0.7 and 1 (GeV/c)^2< Q^2< 17 (GeV/c)^2, we obtain $\int^{0.7}_{0.014} g^n_1(x)dx = -0.036 \pm 0.004 (stat) \pm 0.005 (syst)$ at an average $Q^2=5 (GeV/c)^2$. We find relatively large negative values for $g^n_1$ at low $x$. The results call into question the usual Regge theory method for extrapolating to x=0 to find the full neutron integral $\int^1_0 g^n_1(x)dx$, needed for testing quark-parton model and QCD sum rules.Comment: 5 pages, 3 figures To be published in Phys. Rev. Let

The Sun Health Research Institute Brain Donation Program: Description and Eexperience, 1987–2007

The Brain Donation Program at Sun Health Research Institute has been in continual operation since 1987, with over 1000 brains banked. The population studied primarily resides in the retirement communities of northwest metropolitan Phoenix, Arizona. The Institute is affiliated with Sun Health, a nonprofit community-owned and operated health care provider. Subjects are enrolled prospectively to allow standardized clinical assessments during life. Funding comes primarily from competitive grants. The Program has made short postmortem brain retrieval a priority, with a 2.75-h median postmortem interval for the entire collection. This maximizes the utility of the resource for molecular studies; frozen tissue from approximately 82% of all cases is suitable for RNA studies. Studies performed in-house have shown that, even with very short postmortem intervals, increasing delays in brain retrieval adversely affect RNA integrity and that cerebrospinal fluid pH increases with postmortem interval but does not predict tissue viability

Viterbi decoding of CRES signals in Project 8

Cyclotron radiation emission spectroscopy (CRES) is a modern approach for determining charged particle energies via high-precision frequency measurements of the emitted cyclotron radiation. For CRES experiments with gas within the fiducial volume, signal and noise dynamics can be modelled by a hidden Markov model. We introduce a novel application of the Viterbi algorithm in order to derive informational limits on the optimal detection of cyclotron radiation signals in this class of gas-filled CRES experiments, thereby providing concrete limits from which future reconstruction algorithms, as well as detector designs, can be constrained. The validity of the resultant decision rules is confirmed using both Monte Carlo and Project 8 data

Современные показатели распространенности бронхиальной астмы среди детей

An investigation of bronchial asthma spread according to the ISAAC programme was performed. Under the data obtained the spread of bronchial asthma symptoms was 20.52% among the 1st class pupils and 22.17% among the 8th class pupils. As an examination result bronchial asthma was detected in 7.8% of the 1st class pupils and 8.3% of the 8th class pupils. These values exceed significantly official data of childhood bronchial asthma spread. Bronchial asthma was diagnosed considerably more frequent in children living in wet and mouldy buildings. Elderly children were noted to have a correlation between bronchial asthma and passive smoking. A reliable relation was shown between bronchial asthma in younger children and a presence of home dust collectors, such as carpets, at a child’s bedroom.Проведено исследование распространенности бронхиальной астмы по программе ISAAC. Согласно полученным данным, уровень распространенности «симптомов бронхиальной астмы» у учеников 1-х классов составил 20,52%, а у учеников 8-х классов — 22,17%. При проведении обследования бронхиальная астма была выявлена у 7,8% учеников первых классов и 8,3% школьников 8-х классов. Эти цифры значительно превышают официальные данные распространенности бронхиальной астмы у детей. Диагноз бронхиальной астмы достоверно чаще отмечался у детей, проживавших в помещениях с повышенной влажностью, наличием плесени. У детей старшей возрастной группы была отмечена связь бронхиальной астмы с фактором пассивного курения. Отмечена достоверная связь бронхиальной астмы у детей младшей возрастной группы с присутствием коллекторов домашней пыли (ковров) в спальне ребенка

Consensus guidelines for the use and interpretation of angiogenesis assays

The formation of new blood vessels, or angiogenesis, is a complex process that plays important roles in growth and development, tissue and organ regeneration, as well as numerous pathological conditions. Angiogenesis undergoes multiple discrete steps that can be individually evaluated and quantified by a large number of bioassays. These independent assessments hold advantages but also have limitations. This article describes in vivo, ex vivo, and in vitro bioassays that are available for the evaluation of angiogenesis and highlights critical aspects that are relevant for their execution and proper interpretation. As such, this collaborative work is the first edition of consensus guidelines on angiogenesis bioassays to serve for current and future reference

MEASUREMENT AND QUANTITATIVE ANALYSIS OF DRUG TRANSPORT IN A 3-DIMENSIONAL IN VITRO MODEL OF THE BLOOD-BRAIN BARRIER

The kinetics of solute transport across cell monolayers is complex, and often consists of multiple active transport processes in addition to passive diffusion. The transwell assay is widely used as an in vitro tool to assess transport of small molecules and potential therapeutics across biological barriers. In this assay a monolayer of cells is grown on a porous membrane that separates a top (apical) chamber from a bottom (basolateral) chamber. The molecule of interest is introduced into the apical chamber, and the apparent permeability Papp is calculated from the time-dependent concentration in the basolateral chamber. The transwell assay provides a quantitative measurement of solute permeability, and it is widely used with monolayers of epithelial and endothelial cells to model brain penetration and absorption of intravenously or orally administered drugs. Permeability is a global measure of transport rate but provides little insight into the details of the operating mechanisms. The purpose of this study is to demonstrate that mechanistic details of transport across biological barriers can be obtained from live cell imaging in a perfusable microvessel model with physiologically relevant geometry. Specifically, we report on the kinetics of doxorubicin transport in microvessels formed from three Madin-Darby Canine Kidney (MDCK) epithelial cell lines. MDCK cells form tight junctions, thereby minimizing paracellular transport. One of the cell lines used are genetically engineered versions of MDCKs that are routinely used to screen for substrates of the P-gp efflux pump, one of the main multidrug efflux transporters in brain microvascular endothelial cells