APLF (C2orf13) is a novel human protein involved in the cellular response to chromosomal DNA strand breaks

Aprataxin and polynucleotide kinase (PNK) are DNA end processing factors that are recruited into the DNA single- and double-strand break repair machinery through phosphorylation-specific interactions with XRCC1 and XRCC4, respectively. These interactions are mediated through a divergent class of forkhead-associated (FHA) domain that binds to peptide sequences in XRCC1 and XRCC4 that are phosphorylated by casein kinase 2 (CK2). Here, we identify the product of the uncharacterized open reading frame C2orf13 as a novel member of this FHA domain family of proteins and we denote this protein APLF (aprataxin- and PNK-like factor). We show that APLF interacts with XRCC1 in vivo and in vitro in a manner that is stimulated by CK2. Yeast two-hybrid analyses suggest that APLF also interacts with the double-strand break repair proteins XRCC4 and XRCC5 (Ku86). We also show that endogenous and yellow fluorescent protein-tagged APLF accumulates at sites of H(2)O(2) or UVA laser-induced chromosomal DNA damage and that this is achieved through at least two mechanisms: one that requires the FHA domain-mediated interaction with XRCC1 and a second that is independent of XRCC1 but requires a novel type of zinc finger motif located at the C terminus of APLF. Finally, we demonstrate that APLF is phosphorylated in a DNA damage- and ATM-dependent manner and that the depletion of APLF from noncycling human SH-SY5Y neuroblastoma cells reduces rates of chromosomal DNA strand break repair following ionizing radiation. These data identify APLF as a novel component of the cellular response to DNA strand breaks in human cells

Endoscopic Unroofing of a Choledochocele

A 42-year-old man with previous laparoscopic cholecystectomy was referred for further evaluation of recurrent acute pancreatitis. Secretin-enhanced magnetic resonance cholangiopancreatography showed a 16 mm × 11 mm T2 hyperintense cystic lesion at the major papilla (Figure 1). Upper endoscopic ultrasound (EUS) showed a 15 mm × 10 mm oval, intramural, subepithelial lesion at the major papilla (Figure 2). Endoscopic retrograde cholangiopancreatography (ERCP) showed an 18-mm bulging lesion at the major papilla with normal overlying mucosa (Figure 3); injected contrast collected into a 16-mm cystic cavity (Figure 4). Findings were suggestive of type A choledochocele. A 10–12-mm freehand precut papillotomy was made with a monofilament needle-knife (Huibregtse Single-Lumen Needle Knife, Cook Medical, Bloomington, IN) using an ERBE VIO electrocautery system (ERBE USA; Marietta, GA). The incision was made as long as safely possible in an attempt to open the choledochocele completely and thus expose its walls and contents. We used a standard pull sphincterotome and ERBE electrocautery to perform the pancreatic sphincterotomy, followed by placement of a pancreatic stent. Biliary sphincterotomy was performed using the same technique (settings for needle-knife and pull sphincterotomies: Endocut I, blend current, effect 2/duration 2/interval 3). Biopsies of the inverted choledochocele showed biliary mucosa and duodenal columnar epithelium with inflammation and fibrosis, and no dysplasia. Follow-up ERCP at 4 weeks showed adequate unroofing of the choledochocele (Figure 5); the pancreatic stent was subsequently removed. The patient reported no recurrence of acute pancreatitis at 6-, 12-, and 18-month follow-up intervals

Pion-proton scattering and isospin breaking in the Δ0−Δ++\Delta^0-\Delta^{++} system

We determine the mass and width of the $\Delta^{++}\ (\Delta^0)$ resonance from data on $\pi^+ p\ (\pi^- p)$ scattering both, in the pole of the $S$-matrix and conventional Breit-Wigner approaches to the scattering amplitude. We provide a simple formula that relates the two definitions for the parameters of the $\Delta$. Isospin symmetry breaking in the \d0-\dm system depends on the definition of the resonant properties: we find $M_0-M_{++} = 0.40 \pm 0.57\ {\rm MeV},\ \Gamma_0 -\Gamma_{++} = 6.89 \pm 0.95\ {\rm MeV}$ in the pole approach while $\wt{M}_0-\wt{M}_{++} = 2.25 \pm 0.68\ {\rm MeV},\ \wt{\Gamma}_0 - \wt{\Gamma}_{++} = 8.45 \pm 1.11\ {\rm MeV}$ in the conventional approach.Comment: Latex, 23 pages, two figures upon reques

Physical mapping integrated with syntenic analysis to characterize the gene space of the long arm of wheat chromosome 1A

Background: Bread wheat (Triticum aestivum L.) is one of the most important crops worldwide and its production faces pressing challenges, the solution of which demands genome information. However, the large, highly repetitive hexaploid wheat genome has been considered intractable to standard sequencing approaches. Therefore the International Wheat Genome Sequencing Consortium (IWGSC) proposes to map and sequence the genome on a chromosome-by-chromosome basis. Methodology/Principal Findings: We have constructed a physical map of the long arm of bread wheat chromosome 1A using chromosome-specific BAC libraries by High Information Content Fingerprinting (HICF). Two alternative methods (FPC and LTC) were used to assemble the fingerprints into a high-resolution physical map of the chromosome arm. A total of 365 molecular markers were added to the map, in addition to 1122 putative unique transcripts that were identified by microarray hybridization. The final map consists of 1180 FPC based or 583 LTC based contigs. Conclusions/Significance: The physical map presented here marks an important step forward in mapping of hexaploid bread wheat. The map is orders of magnitude more detailed than previously available maps of this chromosome, and the assignment of over a thousand putative expressed gene sequences to specific map locations will greatly assist future functional studies. This map will be an essential tool for future sequencing of and positional cloning within chromosome 1A

Quantitative analysis of gene expression changes in response to genotoxic compounds

Techniques that quantify molecular endpoints sufficiently sensitive to identify and classify potentially toxic compounds have wide potential for high-throughput in vitro screening. Expression of three genes, RAD51C, TP53 and cystatin A (CSTA), in HEPG2 cells was measured by Q-PCR amplification. In parallel, we developed alternative assays for the same 3 gene signature based on an acridinium-ester chemiluminescent reporter molecule. HEPG2 cells were challenged with eighteen different compounds (n = 18) chosen to represent compounds that are genotoxic (n = 8), non-genotoxic non-carcinogenic (n = 2) or have a less well defined mechanism of action with respect to genotoxicity (n = 8). At least one of the three genes displayed dysregulated expression in the majority of compounds tested by Q-PCR and ten compounds changed the CSTA expression significantly. Acridinium-ester labelled probes for the three genes were synthesised and tested. Analytical sensitivity was characterised and suggested a limit of detection generally better than 0.1 fmol but often 10–50 attomol. A linear amplification step was optimised and this quantitative method detected statistically significant increases in RAD51C and CSTA expression in agreement with the Q-PCR results, demonstrating the potential of this technology. The broad agreement of the amplified chemiluminescent method and Q-PCR in measuring gene expression suggests wider potential application for this chemiluminescent technology

Endoscopic retrieval of a proximally migrated biliary stent: digital cholangioscope to the rescue

Endoscopic techniques for the retrieval of proximally migrated biliary stents include the following: fluoroscopy-guided grasping of the stent with a rat-tooth forceps, balloon placement parallel to the stent with traction retrieval, cannulation of the stent lumen with a wire (standard technique, or use of the curved plastic tip of a modified Soehendra stent retriever [Cook Medical, Bloomington, Ind]) followed by use of standard or modified Soehendra stent retriever, wire-guided retrieval basket, and snare. The technique used depends on the extent of proximal stent migration, the presence of ductal dilatation or biliary stricture, and the endoscopist’s experience. This report describes the retrieval of a proximally migrated biliary stent within an aberrant right hepatic duct (RHD) using a digital cholangioscope (SpyGlass DS system; Boston Scientific, Natick, Mass

Annular pancreas: endoscopic and pancreatographic findings from a tertiary referral ERCP center

Background and Aims Annular pancreas is a congenital anomaly whereby pancreatic tissue encircles the duodenum. Current knowledge of endoscopic findings of annular pancreas is limited to small case series. The aim of this study was to describe the endoscopic and pancreatographic findings of patients with annular pancreas at a large tertiary care ERCP center. Methods This is a retrospective observational study. Our Institutional Review Board–approved, prospectively collected ERCP database was queried for cases of annular pancreas. The electronic medical records were searched for patient and procedure-related data. Results From January 1, 1994, to December 31, 2016, 46 patients with annular pancreas underwent ERCP at our institution. Index ERCP was technically successful in 42 patients (91.3%), and technical success was achieved in all 46 patients (100%) after 2 attempts, when required. A duodenal narrowing or ring was found in most patients (n = 39, 84.8%), yet only 2 (4.3%) had retained gastric contents. Pancreas divisum was found in 21 patients (45.7%), 18 of which were complete divisum. Pancreatobiliary neoplasia was the indication for ERCP in 7 patients (15.2%). Pancreatographic findings consistent with chronic pancreatitis were noted in 15 patients (32.6%) at the index ERCP. Conclusion This is the largest series describing the endoscopic and pancreatographic findings of patients with annular pancreas. We found that 45.7% of patients had concurrent pancreas divisum. Endoscopic therapy was successful in most patients at our institution after 1 ERCP, and in all patients after a second ERCP. Nearly one-third of patients had findings consistent with chronic pancreatitis at the time of index ERCP. It is unclear whether this may be a feature of the natural history of annular pancreas

Dampers for Stationary Labyrinth Seals

Vibration dampers have been invented that are incorporated as components within the stationary labyrinth seal assembly. These dampers are intended to supplement other vibration-suppressing features of labyrinth seals in order to reduce the incidence of high-cycle-fatigue failures, which have been known to occur in the severe vibratory environments of jet engines and turbopumps in which labyrinth seals are typically used. A vibration damper of this type includes several leaf springs and/or a number of metallic particles (shot) all held in an annular seal cavity by a retaining ring. The leaf springs are made of a spring steel alloy chosen, in conjunction with design parameters, to maintain sufficient preload to ensure effectiveness of damping at desired operating temperatures. The cavity is vented via a small radial gap between the retaining ring and seal housing. The damping mechanism is complex. In the case of leaf springs, the mechanism is mainly friction in the slippage between the seal housing and individual dampers. In the case of a damper that contains shot, the damping mechanism includes contributions from friction between individual particles, friction between particles and cavity walls, and dissipation of kinetic energy of impact. The basic concept of particle/shot vibration dampers has been published previously; what is new here is the use of such dampers to suppress traveling-wave vibrations in labyrinth seals. Damping effectiveness depends on many parameters, including, but not limited to, coefficient of friction, mode shape, and frequency and amplitude of vibrational modes. In tests, preloads of the order of 6 to 15 lb (2.72 to 6.8 kilograms) per spring damper were demonstrated to provide adequate damping levels. Effectiveness of shot damping of vibrations having amplitudes from 20 to 200 times normal terrestrial gravitational acceleration (196 to 1,960 meters per square second) and frequencies up to 12 kHz was demonstrated for shot sizes from 0.032 to 0.062 in. (0.8 to 1.6 millimeters) at fill levels of from 70 to 95 percent