Resistance of Human Cytomegalovirus to Cyclopropavir Maps to a Base Pair Deletion in the Open Reading Frame of \u3cem\u3eUL97\u3c/em\u3e

Human cytomegalovirus (HCMV) is a widespread pathogen in the human population, affecting many immunologically immature and immunocompromised patients, and can result in severe complications, such as interstitial pneumonia and mental retardation. Current chemotherapies for the treatment of HCMV infections include ganciclovir (GCV), foscarnet, and cidofovir. However, the high incidences of adverse effects (neutropenia and nephrotoxicity) limit the use of these drugs. Cyclopropavir (CPV), a guanosine nucleoside analog, is 10-fold more active against HCMV than GCV (50% effective concentrations [EC50s] = 0.46 and 4.1 μM, respectively). We hypothesize that the mechanism of action of CPV is similar to that of GCV: phosphorylation to a monophosphate by viral pUL97 protein kinase with further phosphorylation to a triphosphate by endogenous kinases, re- sulting in inhibition of viral DNA synthesis. To test this hypothesis, we isolated a CPV-resistant virus, sequenced its genome, and discovered that bp 498 of UL97 was deleted. This mutation caused a frameshift in UL97 resulting in a truncated protein that lacks a kinase domain. To determine if this base pair deletion was responsible for drug resistance, the mutation was engineered into the wild-type viral genome, which was then exposed to increasing concentrations of CPV. The results demonstrate that the engineered virus was approximately 72-fold more resistant to CPV (EC50 = 25.8 ± 3.1 μM) than the wild-type virus (EC50 = 0.36 ± 0.11 μM). We conclude, therefore, that this mutation is sufficient for drug resistance and that pUL97 is involved in the mechanism of action of CPV

Haemorrhagic Colitis Associated with Enterohaemorrhagic \u3ci\u3eEscherichia coli\u3c/i\u3e O165:H25 Infection in a Yearling Feedlot Heifer

Introduction: Enterohaemorrhagic Escherichia coli (EHEC) cause haemorrhagic colitis and haemolytic uraemic syndrome in humans. Although EHEC infection typically results in haemorrhagic colitis in all ages of human patients, in cattle it is usually limited to 1- to 5-week-old nursing calves. Case Presentation: A 1-year-old feedlot beef heifer was moribund with neurological signs and bloody diarrhoea. At necropsy, the colonic mucosa contained multiple grossly visible haemorrhagic erosions, each measuring \u3c1 mm in diameter. Histologically, foci corresponding to the gross erosions had E. coli O165 antigen-positive bacterial rods adherent to the apical surfaces of degenerate and necrotic colonic mucosal epithelial cells in association with attaching and effacing lesions, and also within cytoplasmic vacuoles in some of these cells. An E. coli O165:H25 strain was isolated from the colonic mucosal tissue, and by microarray analysis was found to contain virulence genes corresponding to type III secretion system (T3SS) structure and regulation (cesD, cesT, escD, escF, escN/escV, escR, escT, ler, sepL, sepQ), T3SS effectors (espA, espB, espC, espD, espD, espF, espH, espJ, nleB, nleC, nleD, nleH, tir), serine proteases (eatA, espC, espP), Shiga toxin (stx2), EHEC-haemolysin (ehxA), and adhesins [intimin-ε (eae-ε), type 1 fimbria (fimA, fimB, fimH), type IV pili (pilA, pilB, pilC, pilM, pilP, pilQ) and non-fimbrial adhesin (efa1/lifA)]. Conclusion: To the best of our knowledge, this is the first report of disease in cattle associated with EHEC O165:H25 infection, the oldest bovine EHEC disease case with isolation of the pathogen and the first bovine case to demonstrate grossly evident, haemorrhagic, colonic mucosal erosions associated with EHEC infection

Haemorrhagic Colitis Associated with Enterohaemorrhagic \u3ci\u3eEscherichia coli\u3c/i\u3e O165:H25 Infection in a Yearling Feedlot Heifer

Introduction: Enterohaemorrhagic Escherichia coli (EHEC) cause haemorrhagic colitis and haemolytic uraemic syndrome in humans. Although EHEC infection typically results in haemorrhagic colitis in all ages of human patients, in cattle it is usually limited to 1- to 5-week-old nursing calves. Case Presentation: A 1-year-old feedlot beef heifer was moribund with neurological signs and bloody diarrhoea. At necropsy, the colonic mucosa contained multiple grossly visible haemorrhagic erosions, each measuring \u3c1 mm in diameter. Histologically, foci corresponding to the gross erosions had E. coli O165 antigen-positive bacterial rods adherent to the apical surfaces of degenerate and necrotic colonic mucosal epithelial cells in association with attaching and effacing lesions, and also within cytoplasmic vacuoles in some of these cells. An E. coli O165:H25 strain was isolated from the colonic mucosal tissue, and by microarray analysis was found to contain virulence genes corresponding to type III secretion system (T3SS) structure and regulation (cesD, cesT, escD, escF, escN/escV, escR, escT, ler, sepL, sepQ), T3SS effectors (espA, espB, espC, espD, espD, espF, espH, espJ, nleB, nleC, nleD, nleH, tir), serine proteases (eatA, espC, espP), Shiga toxin (stx2), EHEC-haemolysin (ehxA), and adhesins [intimin-ε (eae-ε), type 1 fimbria (fimA, fimB, fimH), type IV pili (pilA, pilB, pilC, pilM, pilP, pilQ) and non-fimbrial adhesin (efa1/lifA)]. Conclusion: To the best of our knowledge, this is the first report of disease in cattle associated with EHEC O165:H25 infection, the oldest bovine EHEC disease case with isolation of the pathogen and the first bovine case to demonstrate grossly evident, haemorrhagic, colonic mucosal erosions associated with EHEC infection

Investigation of International Space Station Major Constituent Analyzer Anomalous ORU 02 Performance

The Major Constituent Analyzer (MCA) is a mass spectrometer-based instrument designed to provide critical monitoring of six major atmospheric constituents; nitrogen, oxygen, hydrogen, carbon dioxide, methane, and water vapor on-board the International Space Station. It is an integral part of the Environmental Control and Life Support System (ECLSS). The MCA system is comprised of seven orbit-replaceable units (ORUs) that provide flexibility in maintaining the MCA. Of these, ORU 02, the analyzer assembly requires replacement every 1 to 2 years due to the consumption of limited life components including the ion pump and ion source filaments. Typically, ORU 02s that reach end of life are swapped out of the MCA on orbit and replaced with the on-orbit spare. The replaced ORU 02 is then returned to the OEM for refurbishment and is then return to service. Recently, 2 refurbished ORU 02s, serial numbers F0001 and F0003, failed on orbit shortly after being installed into the MCA. Both ORU 02s have been returned to ground for TT&E, and a failure investigation is underway. The failure signatures have been reproduced on the ground and an initial investigation has determined that both ORU 02 failures involve either the ion source or the ion source control electronics. This paper discusses the results of the failure investigation, the steps required to refurbish the ORU 02s, and the risk mitigation steps that are being incorporated into the refurbishment process to preclude the reoccurrence of these failures in the futur

Investigation of International Space Station Major Constituent Analyzer Anomalous ORU 02 Performance

The Major Constituent Analyzer (MCA) is a mass spectrometer based system that measures the major atmospheric constituents on the International Space Station. In 2011, two MCA ORU 02 analyzer assemblies experienced premature on-orbit failures. These failures were determined to be the result of off-nominal ion source filament performance. Recent product improvements to ORU 02 designed to improve the lifetime of the ion pump also constrained the allowable tuning criteria for the ion source filaments. This presentation describes the filament failures as well as the corrective actions implemented to preclude such failures in the future

The economic impact of Los Alamos National Laboratory on north-central New Mexico and the state of New Mexico fiscal year 1998

Los Alamos National Laboratory (LANL) is a multidisciplinary, multiprogram laboratory with a mission to enhance national military and economic security through science and technology. Its mission is to reduce the nuclear danger through stewardship of the nation`s nuclear stockpile and through its nonproliferation and verification activities. An important secondary mission is to promote US industrial competitiveness by working with US companies in technology transfer and technology development partnerships. Los Alamos is involved in partnerships and collaborations with other federal agencies, with industry (including New Mexico businesses), and with universities worldwide. For this report, the reference period is FY 1998 (October 1, 1997, through September 30, 1998). It includes two major impact analysis: the impact of LANL activities on north-central New Mexico and the economic impacts of LANL on the state of New Mexico. Total impact represents both direct and indirect responding by business, including induced effects (responding by households). The standard multipliers used in determining impacts result from the inter-industry, input-output models developed for the three-county region and the state of New Mexico

Investigation into the High Voltage Shutdown of the Oxygen Generator System in the International Space Station

The Oxygen Generation System (OGS) Hydrogen Dome Assembly Orbital Replacement Unit (ORU) serial number 00001 suffered a cell stack high-voltage shutdown on July 5, 2010. The Hydrogen Dome Assembly ORU was removed and replaced with the on-board spare ORU serial number 00002 to maintain OGS operation. The Hydrogen Dome Assembly ORU was returned from ISS on STS-133/ULF-5 in March 2011 with test, teardown and evaluation (TT&E) and failure analysis to follow

Discovery and validation of serum glycoprotein biomarkers for high grade serous ovarian cancer

Purpose: This study aimed to identify serum glycoprotein biomarkers for early detection of high-grade serous ovarian cancer (HGSOC), the most common and aggressive histotype of ovarian cancer./ Experimental design: The glycoproteomics pipeline lectin magnetic bead array (LeMBA)-mass spectrometry (MS) was used in age-matched case-control serum samples. Clinical samples collected at diagnosis were divided into discovery (n = 30) and validation (n = 98) sets. We also analysed a set of preclinical sera (n = 30) collected prior to HGSOC diagnosis in the UK Collaborative Trial of Ovarian Cancer Screening./ Results: A 7-lectin LeMBA-MS/MS discovery screen shortlisted 59 candidate proteins and three lectins. Validation analysis using 3-lectin LeMBA-multiple reaction monitoring (MRM) confirmed elevated A1AT, AACT, CO9, HPT and ITIH3 and reduced A2MG, ALS, IBP3 and PON1 glycoforms in HGSOC. The best performing multimarker signature had 87.7% area under the receiver operating curve, 90.7% specificity and 70.4% sensitivity for distinguishing HGSOC from benign and healthy groups. In the preclinical set, CO9, ITIH3 and A2MG glycoforms were altered in samples collected 11.1 ± 5.1 months prior to HGSOC diagnosis, suggesting potential for early detection./ Conclusions and clinical relevance: Our findings provide evidence of candidate early HGSOC serum glycoprotein biomarkers, laying the foundation for further study in larger cohorts

Structural and Biochemical Characterization of CIB1 Delineates a New Family of EF-hand-containing Proteins

CIB1 (CIB) is an EF-hand-containing protein that binds multiple effector proteins, including the platelet alphaIIbbeta3 integrin and several serine/threonine kinases and potentially modulates their function. The crystal structure for Ca(2+)-bound CIB1 has been determined at 2.0 A resolution and reveals a compact alpha-helical protein containing four EF-hands, the last two of which bind calcium ions in the standard fashion seen in many other EF-hand proteins. CIB1 shares high structural similarity with calcineurin B and the neuronal calcium sensor (NCS) family of EF-hand-containing proteins. Most importantly, like calcineurin B and NCS proteins, which possess a large hydrophobic pocket necessary for ligand binding, CIB1 contains a hydrophobic pocket that has been implicated in ligand binding by previous mutational analysis. However, unlike several NCS proteins, Ca(2+)-bound CIB1 is largely monomeric whether bound to a relevant peptide ligand or ligand-free. Differences in structure, oligomeric state, and phylogeny define a new family of CIB1-related proteins that extends from arthropods to humans

The economic impact of the Department of Energy on the state of New Mexico fiscal year 1997

The US Department of Energy (DOE) provides a major source of economic benefits in New Mexico. The agency`s far-reaching economic influence within the state is the focus of this report. Economic benefits arising from the various activities and functions of both DOE and its contractors have accrued to the state continuously for over 50 years. For several years, DOE/Albuquerque Operations Office (AL) and New Mexico State University (NMSU) have maintained inter-industry, input-output modeling capabilities to assess DOE`s impacts on the state of New Mexico and the other substate regions most directly impacted by DOE activities. One of the major uses of input-output techniques is to assess the effects of developments initiated outside the economy such as federal DOE monies that flow into the state, on an economy. The information on which the models are based is updated periodically to ensure the most accurate depiction possible of the economy for the period of reference. For this report, the reference periods are Fiscal Year (FY) 1996 and FY 1997. Total impacts represents both direct and indirect impacts (respending by business), including induced (respending by households) effects. The standard multipliers used in determining impacts result from the inter-industry, input-output models uniquely developed for New Mexico. This report includes seven main sections: (1) introduction; (2) profile of DOE activities in New Mexico; (3) DOE expenditure patterns; (4) measuring DOE/New Mexico`s economic impact; (5) technology transfer within the federal labs funded by DOE/New Mexico; (6) glossary of terms; and (7) technical appendix containing a description of the model. 9 figs., 19 tabs