Tackling the Mouse‐on‐Mouse Problem in Cochlear Immunofluorescence: A Simple Double‐Blocking Protocol for Immunofluorescent Labeling of Murine Cochlear Sections with Primary Mouse Antibodies

The mouse is the most widely used animal model in hearing research. Immunohistochemistry and immunofluorescent staining of murine cochlear sections have, thus, remained a backbone of inner ear research. Since many primary antibodies are raised in mouse, the problem of "mouse-on-mouse" background arises due to the interaction between the anti-mouse secondary antibody and the native mouse immunoglobulins. Here, we describe the pattern of mouse-on-mouse background fluorescence in sections of the postnatal mouse cochlea. Furthermore, we describe a simple double-blocking immunofluorescence protocol to label mouse cochlear cryosections. The protocol contains a conventional blocking step with serum, and an additional blocking step with a commercially available anti-mouse IgG blocking reagent. This blocking technique virtually eliminates the "mouse-on-mouse" background in murine cochlear sections, while adding only a little time to the staining protocol. We provide detailed instructions and practical tips for tissue harvesting, processing, and immunofluorescence-labeling. Further protocol modifications are described, to shorten the duration of the protocol, based on the primary antibody incubation temperature. Finally, we demonstrate examples of immunofluorescence staining performed using different incubation times and various incubation temperatures with a commercially available mouse monoclonal primary antibody

Ecohealth trainer manual

This training manual, and the Field Building Leadership Initiative (FBLI) of which it is one component, is part of a global initiative to build capacity in ecosystem approaches to health. Although several books and journals provide materials for learners about Ecohealth, the FBLI Ecohealth Trainer Manual is intended primarily for lecturers, teachers, and trainers. The focus here is on how to teach Ecohealth, providing teachers and trainers with a starting point from which to explore, improvise, adapt, and develop diverse educational Ecohealth learning experiences for and with their participants

Genome wide association mapping of grain arsenic, copper, molybdenum and zinc in rice (Oryza sativa L.) grown at four international field sites

Peer reviewedPublisher PD

Graphene oxide modified with PMMA via ATRP as a reinforcement filler

Graphene is a two-dimensional new allotrope of carbon, which is stimulating great curiosity due to its superior mechanical, electrical, thermal and optical properties. Particularly attractive is the availability of bulk quantities of graphene (G) which can be easily processed by chemical exfoliation, yielding graphene oxide (GO). The resultant oxygenated graphene sheets covered with hydroxyl, epoxy and carboxyl groups offer tremendous opportunities for further functionalization opening plenty of opportunities for the preparation of advanced composite materials. In this work poly(methyl methacrylate) (PMMA) chains have been grafted from the GO surface via atom transfer radical polymerization (ATRP), yielding a nanocomposite which was soluble in chloroform. The surface of the PMMA grafted GO (GPMMA) was characterized by AFM, HRTEM, Raman, FTIR and contact angle. The interest of these novel nanocomposites lies in their potential to be homogenously dispersed in polymeric dense matrices and to promote good interfacial adhesion, of particular relevance in stress transfer to the fillers. PMMA composite films were prepared using different percentages of GPMMA and pristine GO. Mechanical analysis of the resulting films showed that loadings as low as 1% (w/w) of GPMMA are effective reinforcing agents, yielding tougher films than pure PMMA films and even than composite films of PMMA prepared with GO. In fact, addition of 1% (w/w) of GPMMA fillers led to a significant improvement of the elongation at break, yielding a much more ductile and therefore tougher material. Thermal analysis showed an increase of the thermal stability properties of these films providing evidence that strong interfacial interactions between PMMA and GPMMA are achieved. In addition, AFM analysis, in friction force mode, is demonstrated to be an effective tool to analyse the surface filler distribution on polymer matrices

JUNO Conceptual Design Report

The Jiangmen Underground Neutrino Observatory (JUNO) is proposed to determine the neutrino mass hierarchy using an underground liquid scintillator detector. It is located 53 km away from both Yangjiang and Taishan Nuclear Power Plants in Guangdong, China. The experimental hall, spanning more than 50 meters, is under a granite mountain of over 700 m overburden. Within six years of running, the detection of reactor antineutrinos can resolve the neutrino mass hierarchy at a confidence level of 3-4$\sigma$, and determine neutrino oscillation parameters $\sin^2\theta_{12}$, $\Delta m^2_{21}$, and $|\Delta m^2_{ee}|$ to an accuracy of better than 1%. The JUNO detector can be also used to study terrestrial and extra-terrestrial neutrinos and new physics beyond the Standard Model. The central detector contains 20,000 tons liquid scintillator with an acrylic sphere of 35 m in diameter. $\sim$17,000 508-mm diameter PMTs with high quantum efficiency provide $\sim$75% optical coverage. The current choice of the liquid scintillator is: linear alkyl benzene (LAB) as the solvent, plus PPO as the scintillation fluor and a wavelength-shifter (Bis-MSB). The number of detected photoelectrons per MeV is larger than 1,100 and the energy resolution is expected to be 3% at 1 MeV. The calibration system is designed to deploy multiple sources to cover the entire energy range of reactor antineutrinos, and to achieve a full-volume position coverage inside the detector. The veto system is used for muon detection, muon induced background study and reduction. It consists of a Water Cherenkov detector and a Top Tracker system. The readout system, the detector control system and the offline system insure efficient and stable data acquisition and processing.Comment: 328 pages, 211 figure

KLK3 SNP-SNP interactions for prediction of prostate cancer aggressiveness.

Risk classification for prostate cancer (PCa) aggressiveness and underlying mechanisms remain inadequate. Interactions between single nucleotide polymorphisms (SNPs) may provide a solution to fill these gaps. To identify SNP-SNP interactions in the four pathways (the angiogenesis-, mitochondria-, miRNA-, and androgen metabolism-related pathways) associated with PCa aggressiveness, we tested 8587 SNPs for 20,729 cases from the PCa consortium. We identified 3 KLK3 SNPs, and 1083 (P < 3.5 × 10-9) and 3145 (P < 1 × 10-5) SNP-SNP interaction pairs significantly associated with PCa aggressiveness. These SNP pairs associated with PCa aggressiveness were more significant than each of their constituent SNP individual effects. The majority (98.6%) of the 3145 pairs involved KLK3. The 3 most common gene-gene interactions were KLK3-COL4A1:COL4A2, KLK3-CDH13, and KLK3-TGFBR3. Predictions from the SNP interaction-based polygenic risk score based on 24 SNP pairs are promising. The prevalence of PCa aggressiveness was 49.8%, 21.9%, and 7.0% for the PCa cases from our cohort with the top 1%, middle 50%, and bottom 1% risk profiles. Potential biological functions of the identified KLK3 SNP-SNP interactions were supported by gene expression and protein-protein interaction results. Our findings suggest KLK3 SNP interactions may play an important role in PCa aggressiveness