The specificity and patterns of staining in human cells and tissues of p16INK4a antibodies demonstrate variant antigen binding

The validity of the identification and classification of human cancer using antibodies to detect biomarker proteins depends upon antibody specificity. Antibodies that bind to the tumour-suppressor protein p16INK4a are widely used for cancer diagnosis and research. In this study we examined the specificity of four commercially available anti-p16INK4a antibodies in four immunological applications. The antibodies H-156 and JC8 detected the same 16 kDa protein in western blot and immunoprecipitation tests, whereas the antibody F-12 did not detect any protein in western blot analysis or capture a protein that could be recognised by the H-156 antibody. In immunocytochemistry tests, the antibodies JC8 and H-156 detected a predominately cytoplasmic localised antigen, whose signal was depleted in p16INK4a siRNA experiments. F-12, in contrast, detected a predominately nuclear located antigen and there was no noticeable reduction in this signal after siRNA knockdown. Furthermore in immunohistochemistry tests, F-12 generated a different pattern of staining compared to the JC8 and E6H4 antibodies. These results demonstrate that three out of four commercially available p16INK4a antibodies are specific to, and indicate a mainly cytoplasmic localisation for, the p16INK4a protein. The F-12 antibody, which has been widely used in previous studies, gave different results to the other antibodies and did not demonstrate specificity to human p16INK4a. This work emphasizes the importance of the validation of commercial antibodies, aside to the previously reported use, for the full verification of immunoreaction specificity

From exercise intolerance to functional improvement: The second wind phenomenon in the identification of McArdle disease

McArdle disease is the most common of the glycogen storage diseases. Onset of symptoms is usually in childhood with muscle pain and restricted exercise capacity. Signs and symptoms are often ignored in children or put down to 'growing pains' and thus diagnosis is often delayed. Misdiagnosis is not uncommon because several other conditions such as muscular dystrophy and muscle channelopathies can manifest with similar symptoms. A simple exercise test performed in the clinic can however help to identify patients by revealing the second wind phenomenon which is pathognomonic of the condition. Here a patient is reported illustrating the value of using a simple 12 minute walk test.RSS is funded by Ciências sem Fronteiras/CAPES Foundation. The authors would like to thank the Association for Glycogen Storage Disease (UK), the EUROMAC Registry funded by the European Union, the Muscular Dystrophy Campaign, the NHS National Specialist Commissioning Group and the Myositis Support Group for funding

Boussinesq modeling of a rip current system

In this study, we use a time domain numerical model based on the fully nonlinear extended Boussinesq equations [Wei et al., 1995] to investigate surface wave transformation and breaking-induced nearshore circulation. The energy dissipation due to wave breaking is modeled by introducing an eddy viscosity term into the momentum equations, with the viscosity strongly localized on the front face of the breaking waves. Wave run-up on the beach is simulated using a moving shoreline technique. We employ quasi fourth-order finite difference schemes to solve the governing equations. Satisfactory agreement is found between the numerical results and the laboratory measurements of Haller et al. [1997], including wave height, mean water level, and longshore and cross-shore velocity components. The model results reveal the temporal and spatial variability of the wave-induced nearshore circulation, and the instability of the rip current in agreement with the physical experiment. Insights into the vorticity associated with the rip current and wave diffraction by underlying vortices are obtained

Single-atom imaging of fermions in a quantum-gas microscope

Single-atom-resolved detection in optical lattices using quantum-gas microscopes has enabled a new generation of experiments in the field of quantum simulation. Fluorescence imaging of individual atoms has so far been achieved for bosonic species with optical molasses cooling, whereas detection of fermionic alkaline atoms in optical lattices by this method has proven more challenging. Here we demonstrate single-site- and single-atom-resolved fluorescence imaging of fermionic potassium-40 atoms in a quantum-gas microscope setup using electromagnetically-induced-transparency cooling. We detected on average 1000 fluorescence photons from a single atom within 1.5s, while keeping it close to the vibrational ground state of the optical lattice. Our results will enable the study of strongly correlated fermionic quantum systems in optical lattices with resolution at the single-atom level, and give access to observables such as the local entropy distribution and individual defects in fermionic Mott insulators or anti-ferromagnetically ordered phases.Comment: 7 pages, 5 figures; Nature Physics, published online 13 July 201

Transcriptional profiling of degraded RNA in cryopreserved and fixed tissue samples obtained at autopsy

BACKGROUND: Traditional multiplexed gene expression methods require well preserved, intact RNA. Such specimens are difficult to acquire in clinical practice where formalin fixation is the standard procedure for processing tissue. Even when special handling methods are used to obtain frozen tissue, there may be RNA degradation; for example autopsy samples where degradation occurs both pre-mortem and during the interval between death and cryopreservation. Although specimens with partially degraded RNA can be analyzed by qRT-PCR, these analyses can only be done individually or at low levels of multiplexing and are laborious and expensive to run for large numbers of RNA targets. METHODS: We evaluated the ability of the cDNA-mediated Annealing, Selection, extension, and Ligation (DASL) assay to provide highly multiplexed analyses of cryopreserved and formalin fixed, paraffin embedded (FFPE) tissues obtained at autopsy. Each assay provides data on 1536 targets, and can be performed on specimens with RNA fragments as small as 60 bp. RESULTS: The DASL performed accurately and consistently with cryopreserved RNA obtained at autopsy as well as with RNA extracted from formalin-fixed paraffin embedded tissue that had a cryopreserved mirror image specimen with high quality RNA. In FFPE tissue where the cryopreserved mirror image specimen was of low quality the assay performed reproducibly on some but not all specimens. CONCLUSION: The DASL assay provides reproducible results from cryopreserved specimens and many FFPE specimens obtained at autopsy. Gene expression analyses of these specimens may be especially valuable for the study of non-cancer endpoints, where surgical specimens are rarely available

Icarus Falls: The Coal Health Scandal

The handling of cases under the Coal Health Compensation Schemes, set up in 1999 to compensate miners suffering from workplace medical conditions, resulted in over 100 solicitors from more than 30 firms facing disciplinary proceedings. Three were struck off, three suspended and over forty fined following the largest investigation ever mounted by the regulator. This article examines the political and regulatory context of the scandal, describes one of the cases presented to the Solicitors' Disciplinary Tribunal and examines the relevance of theories of transgression to professional disciplinary matters. It concludes by considering the regulatory impacts and implications of the scandal

Simultaneous high resolution meausurement of phonons and ionization created by particle interactions in a 60 g germanium crystal at 25 mK

We demonstrate simultaneous high energy resolution (rms≊800 eV) measurements of ionization and phonons created by particle interactions in a semiconductor crystal of macroscopic size (60 g germanium) at 25 mK. We present first studies of charge collection at biases below 1 V/cm, and find that, contrary to commonly held opinion, the full recoil energy of particle interactions is recovered as phonons when charge trapping is negligible. We also report an unanticipated correlation between charge collection and phonon energy at very low bias, and discuss this effect in terms of charge trapping

Measurement of ionization and phonon production by nuclear recoils in a 60 g crystal of germanium at 25 mK

We report on the first measurement of the absolute phonon energy and the amount of ionization produced by the recoil of nuclei and electrons in a 60 g germanium cyrstal at a temperature of ≊25 mK. We find good agreement between our results and previous measurements of ionization yield from nuclear recoils in germanium. Our device achieves 10:1 discrimination between neutrons and photons in the few keV energy range, demonstrating the feasibility of this technique for large reductions of background in searches for direct interactions of weakly interacting massive particle dark matter

Association studies of up to 1.2 million individuals yield new insights into the genetic etiology of tobacco and alcohol use.

Tobacco and alcohol use are leading causes of mortality that influence risk for many complex diseases and disorders1. They are heritable2,3 and etiologically related4,5 behaviors that have been resistant to gene discovery efforts6-11. In sample sizes up to 1.2 million individuals, we discovered 566 genetic variants in 406 loci associated with multiple stages of tobacco use (initiation, cessation, and heaviness) as well as alcohol use, with 150 loci evidencing pleiotropic association. Smoking phenotypes were positively genetically correlated with many health conditions, whereas alcohol use was negatively correlated with these conditions, such that increased genetic risk for alcohol use is associated with lower disease risk. We report evidence for the involvement of many systems in tobacco and alcohol use, including genes involved in nicotinic, dopaminergic, and glutamatergic neurotransmission. The results provide a solid starting point to evaluate the effects of these loci in model organisms and more precise substance use measures