Uncoupling of p97 ATPase activity has a dominant negative effect on protein extraction

p97 is a highly abundant, homohexameric AAA+ ATPase that performs a variety of essential cellular functions. Characterized as a ubiquitin-selective chaperone, p97 recognizes proteins conjugated to K48-linked polyubiquitin chains and promotes their removal from chromatin and other molecular complexes. Changes in p97 expression or activity are associated with the development of cancer and several related neurodegenerative disorders. Although pathogenic p97 mutations cluster in and around p97's ATPase domains, mutant proteins display normal or elevated ATPase activity. Here, we show that one of the most common p97 mutations (R155C) retains ATPase activity, but is functionally defective. p97-R155C can be recruited to ubiquitinated substrates on chromatin, but is unable to promote substrate removal. As a result, p97-R155C acts as a dominant negative, blocking protein extraction by a similar mechanism to that observed when p97's ATPase activity is inhibited or inactivated. However, unlike ATPase-deficient proteins, p97-R155C consumes excess ATP, which can hinder high-energy processes. Together, our results shed new insight into how pathogenic mutations in p97 alter its cellular function, with implications for understanding the etiology and treatment of p97-associated diseases

Lesinurad, a novel, oral compound for gout, acts to decrease serum uric acid through inhibition of urate transporters in the kidney.

BackgroundExcess body burden of uric acid promotes gout. Diminished renal clearance of uric acid causes hyperuricemia in most patients with gout, and the renal urate transporter (URAT)1 is important for regulation of serum uric acid (sUA) levels. The URAT1 inhibitors probenecid and benzbromarone are used as gout therapies; however, their use is limited by drug-drug interactions and off-target toxicity, respectively. Here, we define the mechanism of action of lesinurad (Zurampic®; RDEA594), a novel URAT1 inhibitor, recently approved in the USA and Europe for treatment of chronic gout.MethodssUA levels, fractional excretion of uric acid (FEUA), lesinurad plasma levels, and urinary excretion of lesinurad were measured in healthy volunteers treated with lesinurad. In addition, lesinurad, probenecid, and benzbromarone were compared in vitro for effects on urate transporters and the organic anion transporters (OAT)1 and OAT3, changes in mitochondrial membrane potential, and human peroxisome proliferator-activated receptor gamma (PPARγ) activity.ResultsAfter 6 hours, a single 200-mg dose of lesinurad elevated FEUA 3.6-fold (p < 0.001) and reduced sUA levels by 33 % (p < 0.001). At concentrations achieved in the clinic, lesinurad inhibited activity of URAT1 and OAT4 in vitro, did not inhibit GLUT9, and had no effect on ABCG2. Lesinurad also showed a low risk for mitochondrial toxicity and PPARγ induction compared to benzbromarone. Unlike probenecid, lesinurad did not inhibit OAT1 or OAT3 in the clinical setting.ConclusionThe pharmacodynamic effects and in vitro activity of lesinurad are consistent with inhibition of URAT1 and OAT4, major apical transporters for uric acid. Lesinurad also has a favorable selectivity and safety profile, consistent with an important role in sUA-lowering therapy for patients with gout

Well-differentiated papillary mesothelioma of the peritoneum is genetically defined by mutually exclusive mutations in TRAF7 and CDC42.

Well-differentiated papillary mesothelioma is an uncommon mesothelial neoplasm that most frequently arises in the peritoneal cavity of women of reproductive age. Whereas malignant mesothelioma is an aggressive tumor associated with poor outcome, well-differentiated papillary mesothelioma typically exhibits indolent behavior. However, histologically differentiating between these two entities can be challenging, necessitating the development of distinguishing biomarkers. While the genetic alterations that drive malignant mesothelioma have recently been determined, the molecular pathogenesis of well-differentiated papillary mesothelioma is unknown. Here we performed genomic profiling on a cohort of ten well-differentiated papillary mesothelioma of the peritoneum. We identified that all tumors harbored somatic missense mutations in either the TRAF7 or CDC42 genes, and lacked alterations involving BAP1, NF2, CDKN2A, DDX3X, SETD2, and ALK that are frequent in malignant mesothelioma. We recently identified that another mesothelial neoplasm, adenomatoid tumor of the genital tract, is genetically defined by somatic missense mutations in the TRAF7 gene, indicating a shared molecular pathogenesis between well-differentiated papillary mesothelioma and adenomatoid tumors. To the best of our knowledge, well-differentiated papillary mesothelioma is the first human tumor type found to harbor recurrent mutations in the CDC42 gene, which encodes a Rho family GTPase. Immunohistochemistry demonstrated intact BAP1 expression in all cases of well-differentiated papillary mesothelioma, indicating that this is a reliable marker for distinguishing well-differentiated papillary mesothelioma from malignant mesotheliomas that frequently display loss of expression. Additionally, all well-differentiated papillary mesothelioma demonstrated robust expression of L1 cell adhesion molecule (L1CAM), a marker of NF-kB pathway activation, similar to that observed in adenomatoid tumors. In contrast, we have previously shown that L1CAM staining is not observed in normal mesothelial cells and malignant mesotheliomas of the peritoneum. Together, these studies demonstrate that well-differentiated papillary mesothelioma is genetically defined by mutually exclusive mutations in TRAF7 and CDC42 that molecularly distinguish this entity from malignant mesothelioma

Confirmation of a recent bipolar ejection in the very young hierarchical multiple system IRAS 16293-2422

We present and analyze two new high-resolution (approx 0.3 arcsec), high-sensitivity (approx 50 uJy beam-1) Very Large Array 3.6 cm observations of IRAS 16293-2422 obtained in 2007 August and 2008 December. The components A2alpha and A2beta recently detected in this system are still present, and have moved roughly symmetrically away from source A2 at a projected velocity of 30-80 km s-1. This confirms that A2alpha and A2beta were formed as a consequence of a very recent bipolar ejection from A2. Powerful bipolar ejections have long been known to occur in low-mass young stars, but this is -to our knowledge-- the first time that such a dramatic one is observed from its very beginning. Under the reasonable assumption that the flux detected at radio wavelengths is optically thin free-free emission, one can estimate the mass of each ejecta to be of the order of 10^-8 Msun. If the ejecta were created as a consequence of an episode of enhanced mass loss accompanied by an increase in accretion onto the protostar, then the total luminosity of IRAS 16293-2422 ought to have increased by 10-60% over the course of at least several months. Between A2alpha and A2beta, component A2 has reappeared, and the relative position angle between A2 and A1 is found to have increased significantly since 2003-2005. This strongly suggests that A1 is a protostar rather than a shock feature, and that the A1/A2 pair is a tight binary system. Including component B, IRAS 16293-2422 therefore appears to be a very young hierarchical multiple system.Comment: Accepted for publication in The Astrophysical Journa

An endoscopie imaging system based on a two-dimensional CMUT array: real-time imaging results

Real-time catheter-based ultrasound imaging tools are needed for diagnosis and image-guided procedures. The continued development of these tools is partially limited by the difficulty of fabricating two-dimensional array geometries of piezoelectric transducers. Using capacitive micromachined ultrasonic transducer (CMUT) technology, transducer arrays with widely varying geometries, high frequencies, and wide bandwidths can be fabricated. A volumetric ultrasound imaging system based on a two-dimensional, 16×l6-element, CMUT array is presented. Transducer arrays with operating frequencies ranging from 3 MHz to 7.5 MHz were fabricated for this system. The transducer array including DC bias pads measures 4 mm by 4.7 mm. The transducer elements are connected to flip-chip bond pads on the array back side with 400-μm long through-wafer interconnects. The array is flip-chip bonded to a custom-designed integrated circuit (IC) that comprises the front-end electronics. Integrating the front-end electronics with the transducer array reduces the effects of cable capacitance on the transducer's performance and provides a compact means of connecting to the transducer elements. The front-end IC provides a 27-V pulser and 10-MHz bandwidth amplifier for each element of the array. An FPGA-based data acquisition system is used for control and data acquisition. Output pressure of 230 kPa was measured for the integrated device. A receive sensitivity of 125 mV/kPa was measured at the output of the amplifier. Amplifier output noise at 5 Mhz is 112 nV/√Hz. Volumetric images of a wire phantom and vessel phantom are presented. Volumetric data for a wire phantom was acquired in real-time at 30 frames per second.Publisher's Versio

Cost-effective telecom/datacom semiconductor lasers

The recent development of semiconductor laser technologies for cost-effective telecom/datacom applications is reviewed in details in this paper. This includes the laser design, laser chip technology, laser packaging technology and other low cost lasers (chip + packaging). Some design and simulation examples in Archcom laser production are described first. A latest trend in the wafer scale testing/characterization/screening technology for low cost semiconductor laser mass production is discussed then. An advanced long wavelength high power single mode surface emitting laser with wafer scale characterization using our unique mask free focused ion beam (FIB) etching technology is also demonstrated. Detailed descriptions on our wide temperature range (-50 °C to +105 °C) G-PON distributed feedback (DFB) semiconductor lasers with high performance and low cost wafer design are included. Cost reduction innovations in laser package with our beam profile improved laser and optical feedback insensitive (OFBI) laser are also addressed

Molecular testing for the clinical diagnosis of fibrolamellar carcinoma.

Fibrolamellar carcinoma has a distinctive morphology and immunophenotype, including cytokeratin 7 and CD68 co-expression. Despite the distinct findings, accurate diagnosis of fibrolamellar carcinoma continues to be a challenge. Recently, fibrolamellar carcinomas were found to harbor a characteristic somatic gene fusion, DNAJB1-PRKACA. A break-apart fluorescence in situ hybridization (FISH) assay was designed to detect this fusion event and to examine its diagnostic performance in a large, multicenter, multinational study. Cases initially classified as fibrolamellar carcinoma based on histological features were reviewed from 124 patients. Upon central review, 104 of the 124 cases were classified histologically as typical of fibrolamellar carcinoma, 12 cases as 'possible fibrolamellar carcinoma' and 8 cases as 'unlikely to be fibrolamellar carcinoma'. PRKACA FISH was positive for rearrangement in 102 of 103 (99%) typical fibrolamellar carcinomas, 9 of 12 'possible fibrolamellar carcinomas' and 0 of 8 cases 'unlikely to be fibrolamellar carcinomas'. Within the morphologically typical group of fibrolamellar carcinomas, two tumors with unusual FISH patterns were also identified. Both cases had the fusion gene DNAJB1-PRKACA, but one also had amplification of the fusion gene and one had heterozygous deletion of the normal PRKACA locus. In addition, 88 conventional hepatocellular carcinomas were evaluated with PRKACA FISH and all were negative. These findings demonstrate that FISH for the PRKACA rearrangement is a clinically useful tool to confirm the diagnosis of fibrolamellar carcinoma, with high sensitivity and specificity. A diagnosis of fibrolamellar carcinoma is more accurate when based on morphology plus confirmatory testing than when based on morphology alone