Using a general strategy for evaluating clinical tissue specimens, we found that 70% ethanol fixation and paraffin embedding is a useful method for molecular profiling studies. Human prostate and ...kidney were used as test tissues. The protein content of the samples was analyzed by one-dimensional gel electrophoresis, immunoblot, two-dimensional gel electrophoresis, and layered expression scanning. In each case, the fixed and embedded tissues produced results similar to that obtained from snap-frozen specimens, although the protein quantity was somewhat decreased. Recovery of mRNA was reduced in both quantity and quality in the ethanol-fixed samples, but was superior to that obtained from formalin-fixed samples and sufficient to perform reverse transcription polymerase chain reactions. Recovery of DNA from ethanol-fixed specimens was superior to formalin-fixed samples as determined by one-dimensional gel electrophoresis and polymerase chain reaction. In conclusion, specimens fixed in 70% ethanol and embedded in paraffin produce good histology and permit recovery of DNA, mRNA, and proteins sufficient for several downstream molecular analyses. Complete protocols and additional discussion of relevant issues are available on an accompanying website (
http://cgap-mf.nih.gov/).
Vela X is a large, 3
$^{\circ}$
$\times$
2
$^{\circ}$
, radio-emitting pulsar wind nebula (PWN) powered by the Vela pulsar in the Vela supernova remnant. Using four Suzaku/XIS observations pointed ...just outside Vela X, we have found hard X-ray emission extending throughout the fields of view. The hard X-ray spectra are well represented by a power-law. The photon index was measured to be constant at
$\Gamma$
$\sim$
2.4, similar to that of the southern outer part of Vela X. The power-law flux decreases with increasing distance from the pulsar. These properties lead us to propose that the hard X-ray emission is associated with the Vela PWN. The larger X-ray extension found in this work strongly suggests that distinct populations of relativistic electrons form the X-ray PWN and Vela X, as was recently inferred from multiwavelength spectral modeling of Vela X.
Primary congenital glaucoma (PCG) is a leading cause of blindness in children worldwide and is caused by developmental defects in 2 aqueous humor outflow structures, Schlemm's canal (SC) and the ...trabecular meshwork. We previously identified loss-of-function mutations in the angiopoietin (ANGPT) receptor TEK in families with PCG and showed that ANGPT/TEK signaling is essential for SC development. Here, we describe roles for the major ANGPT ligands in the development of the aqueous outflow pathway. We determined that ANGPT1 is essential for SC development, and that Angpt1-knockout mice form a severely hypomorphic canal with elevated intraocular pressure. By contrast, ANGPT2 was dispensable, although mice deficient in both Angpt1 and Angpt2 completely lacked SC, indicating that ANGPT2 compensates for the loss of ANGPT1. In addition, we identified 3 human subjects with rare ANGPT1 variants within an international cohort of 284 PCG patients. Loss of function in 2 of the 3 patient alleles was observed by functional analysis of ANGPT1 variants in a combined in silico, in vitro, and in vivo approach, supporting a causative role for ANGPT1 in disease. By linking ANGPT1 with PCG, these results highlight the importance of ANGPT/TEK signaling in glaucoma pathogenesis and identify a candidate target for therapeutic development.
Annexin I protein expression was evaluated in patient-matched longitudinal study sets of laser capture microdissected normal, premalignant, and invasive epithelium from human esophageal squamous cell ...cancer and prostatic adenocarcinoma. In 25 esophageal cases (20 by Western blot and 5 by immunohistochemistry) and 17 prostate cases (3 by Western blot and 14 by immunohistochemistry), both tumor types showed either complete loss or a dramatic reduction in the level of annexin I protein expression compared with patient-matched normal epithelium (P < or = 0.05). Moreover, by using Western blot analysis of laser capture microdissected, patient-matched longitudinal study sets of both tumor types, the loss of protein expression occurred in premalignant lesions. Concordance of this result with immunohistochemical analysis suggests that annexin I may be an essential component for maintenance of the normal epithelial phenotype. Additional studies investigating the mechanism(s) and functional consequences of annexin I protein loss in tumor cells are warranted.
Purpose This phase 2 study compared OMS103HP (Omeros, Seattle, WA) with control (lactated Ringer's) irrigation solution in patients undergoing arthroscopic partial meniscectomy. Methods This was a ...prospective, multicenter, double-blind, randomized, vehicle-controlled, parallel-group study. Safety and postoperative pain, range of motion, and self-reported function were evaluated for 90 days. Statistical results were based on univariate analysis of variance and repeated-measures analyses. Results Mean visual analog scale (VAS) pain scores within 24 hours after discharge from the recovery room showed more pain in the control group beginning at 2 hours and peaking at 8 hours. Univariate analysis of variance of mean VAS scores over the 24-hour period did not meet statistical significance. Repeated-measures analysis yielded a statistically significant difference ( P = .004) for time-by-treatment interaction, showing a clear drug benefit over time based on VAS scores. There were statistically significant differences at day 7 between the groups in passive flexion without pain ( P = .022). The proportion of patients achieving flexion of 95° or greater, 110°, and 125° was greater for the OMS103HP group. The Knee Injury and Osteoarthritis Outcome Score (KOOS) showed statistically significant differences ( P ≤ .05) between the OMS103HP and control groups for 4 of 5 outcomes (symptoms, pain, sport and recreation, and knee-based quality of life but not activities of daily living). All scores showed a treatment effect through day 90. The overall incidence of adverse events and abnormal laboratory values for the OMS103HP and control groups was similar. Serious adverse events occurred in 1 control patient. Conclusions In this study of patients with meniscal tears who underwent simple debridement, the use of OMS103HP resulted in reduced acute postoperative pain (measured by VAS over the first 24 hours postoperatively), reduced pain during recovery (measured by the KOOS pain subscale, which measures both background levels of pain and exacerbations caused by movements or activities), improved postoperative knee motion, and improved functional outcomes as assessed with the KOOS Knee Survey. Clinical benefits of OMS103HP were consistent and sustained throughout 90 days of postoperative follow-up. Level of Evidence Level I, prospective, randomized, controlled trial.
In the past few years, gamma-ray astronomy has entered a golden age thanks to two major breakthroughs: Cherenkov telescopes on the ground and the Large Area Telescope (LAT) onboard the Fermi ...satellite. The sample of supernova remnants (SNRs) detected at gamma-ray energies is now much larger: it goes from evolved supernova remnants interacting with molecular clouds up to young shell-type supernova remnants and historical supernova remnants. Studies of SNRs are of great interest, as these analyses are directly linked to the long standing issue of the origin of the Galactic cosmic rays. In this context, pulsar wind nebulae (PWNe) need also to be considered since they evolve in conjunction with SNRs. As a result, they frequently complicate interpretation of the gamma-ray emission seen from SNRs and they could also contribute directly to the local cosmic ray spectrum, particularly the leptonic component. This paper reviews the current results and thinking on SNRs and PWNe and their connection to cosmic ray production.
Au cours des dernières années, l'astronomie gamma est entrée dans un âge d'or grâce à deux avancées majeures : les télescopes Tcherenkov au sol et le Large Area Telescope (LAT) à bord du satellite Fermi. L'échantillon des restes de supernova (SNR) détectés en rayons gamma de haute énergie est maintenant beaucoup plus vaste : il va des vestiges de supernovae évolués en interaction avec des nuages moléculaires jusqu'aux jeunes SNR en coquille et aux SNR historiques. Les études des SNR sont d'un grand intérêt, car ces analyses sont directement liées à la question de l'origine des rayons cosmiques galactiques. Dans ce contexte, les nébuleuses de pulsars (PWN) doivent également être prises en compte, car elles évoluent en conjonction avec les SNR. En conséquence, elles compliquent souvent l'interprétation de l'émission gamma en provenance des SNR et pourraient aussi contribuer directement au spectre local de rayons cosmiques, en particulier à sa composante leptonique. Cet article passe en revue les résultats et réflexions actuels concernant les SNR et les PWN, ainsi que leur connexion avec la production des rayons cosmiques.
We present the analysis of Fermi Large Area Telescope gamma -ray observations of HB 21 (G89.0+4.7). We detect significant gamma -ray emission associated with the remnant; the flux > 100 MeV is 9.4 + ...or - 0.8 (stat) + or - 1.6 (syst) x 10 super(-11) erg cm super(-2) s super(-1). HB 21 is well modeled by a uniform disk centered at l = 88degrees.75 + or - 0degrees.04, b = +4degrees.65 + or - 0degrees.06 with a radius of 1degrees.19 + or - 0degrees.06. The gamma -ray spectrum shows clear evidence of curvature, suggesting a cutoff or break in the underlying particle population at an energy of a few GeV. We complement gamma -ray observations with the analysis of the WMAP 7 yr data from 23 to 93 GHz, achieving the first detection of HB 21 at these frequencies. In combination with archival radio data, the radio spectrum shows a special break, which helps to constrain the relativistic electron spectrum, and, in turn, parameters of simple non-thermal radiation models. In one-zone models multi wavelength data favor the origin of gamma rays from nucleon-nucleon collisions. A single population of electrons cannot produce both gamma rays through bremsstrahlung and radio emission through synchrotron radiation. A predominantly inverse-Compton origin of the gamma -ray emission is disfavored because it requires lower interstellar densities than are inferred for HB 21. In the hadronic-dominated scenarios, accelerated nuclei contribute a total energy of ~3 x 10 super(49) erg, while, in a two-zone bremsstrahlung-dominated scenario, the total energy in accelerated particles is ~1 x 10 super(49) erg.
Significant advances are needed to improve the diagnosis, prognosis, and management of persons with CKD. Discovery of new biomarkers and improvements in currently available biomarkers for CKD hold ...great promise to achieve these necessary advances. Interest in identification and evaluation of biomarkers for CKD has increased substantially over the past decade. In 2009, the National Institute of Diabetes and Digestive and Kidney Diseases established the CKD Biomarkers Consortium (http://www.ckdbiomarkersconsortium.org/), a multidisciplinary, collaborative study group located at over a dozen academic medical centers. The main objective of the consortium was to evaluate new biomarkers for purposes related to CKD in established prospective cohorts, including those enriched for CKD. During the first 5 years of the consortium, many insights into collaborative biomarker research were gained that may be useful to other investigators involved in biomarkers research. These lessons learned are outlined in this Special Feature and include a wide range of issues related to biospecimen collection, storage, and retrieval, and the internal and external quality assessment of laboratories that performed the assays. The authors propose that investigations involving biomarker discovery and validation are greatly enhanced by establishing and following explicit quality control metrics, including the use of blind replicate and proficiency samples, by carefully considering the conditions under which specimens are collected, handled, and stored, and by conducting pilot and feasibility studies when there are concerns about the condition of the specimens or the accuracy or reproducibility of the assays.
The ancestors of fungi are believed to be simple aquatic forms with flagellated spores, similar to members of the extant phylum Chytridiomycota (chytrids). Current classifications assume that ...chytrids form an early-diverging clade within the kingdom Fungi and imply a single loss of the spore flagellum, leading to the diversification of terrestrial fungi. Here we develop phylogenetic hypotheses for Fungi using data from six gene regions and nearly 200 species. Our results indicate that there may have been at least four independent losses of the flagellum in the kingdom Fungi. These losses of swimming spores coincided with the evolution of new mechanisms of spore dispersal, such as aerial dispersal in mycelial groups and polar tube eversion in the microsporidia (unicellular forms that lack mitochondria). The enigmatic microsporidia seem to be derived from an endoparasitic chytrid ancestor similar to Rozella allomycis, on the earliest diverging branch of the fungal phylogenetic tree.
Celotno besedilo
Dostopno za:
DOBA, IJS, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
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