Rapid evolution of protein kinase PKR alters sensitivity to viral inhibitors.

Protein kinase PKR (also known as EIF2AK2) is activated during viral infection and phosphorylates the alpha subunit of eukaryotic translation initiation factor 2 (eIF2), leading to inhibition of translation and viral replication. We report fast evolution of the PKR kinase domain in vertebrates, coupled with positive selection of specific sites. Substitution of positively selected residues in human PKR with residues found in related species altered sensitivity to PKR inhibitors from different poxviruses. Species-specific differences in sensitivity to poxviral pseudosubstrate inhibitors were identified between human and mouse PKR, and these differences were traced to positively selected residues near the eIF2alpha binding site. Our findings indicate how an antiviral protein evolved to evade viral inhibition while maintaining its primary function. Moreover, the identified species-specific differences in the susceptibility to viral inhibitors have important implications for studying human infections in nonhuman model systems

Evaluation and optimization of medication related fall risk at a community hospital

Background: Falls occur commonly in older adults at a rate of 3-5 per 1000 beds in healthcare settings. They can results in increased length of stay, morbidity and mortality, and healthcare costs. Some risk factors include older age, environmental hazards and medications. Currently at BHM, patient’s risk for falling is assessed based on Morse Fall Scale (MFS), which does include drugs as part of its assessment. The purpose of this project is to integrate medication risk evaluation into our current fall assessment. Methodology: single-centered, retrospective, IRB-reviewed quality improvement project at BHM. We included patients over age 18 who fell with at least a minor injury (F2 level) in 2019. Our primary outcome was to determine prevalence of common high-fall-risk medications administered in patients who fell and our secondary outcome was to identify pharmacy-led interventions to decrease fall risk. Since MFS does not include medications as part of its score, we then evaluated each patient using a medication scoring system with point value of 1-3 for high-fall-risk medications (point value of 3 is assigned to high-fall-risk medications and point value of 1 is assigned to low fall risk medications). Any patient with a cumulative score of higher than 6 is recognized to be at high fall risk. Results: Primary outcome showed that cardiac agents, followed by opioids, and by benzodiazepines/sleep aids were the highest administered medications in our patients who fell. Based on those findings, we identified 29 opportunities where patient could have received lower doses and/or have had medication scheduling changes in order to reduce fall risk. We are proposing automatic dose adjustments similar to hydromorphone and zolpidem/temazepam protocols where pharmacists can reduce starting doses to lowest dose possible for drug naïve patients. In addition, we are proposing to have medication scoring system to be implemented into our software so pharmacists are notified if a patient is high risk for falls based on MFS score and medication score system. Conclusion: Pharmacists can play a huge role in reducing falls in inpatient settings. Based on our findings, some opportunities where pharmacists can intervene to reduce risk of falling include scheduling and automatic dose reductions with medications such as cardiac agents, opioids, and benzodiazepine/sleep aids

Co-cultivation of murine BMDCs with 67NR mouse mammary carcinoma cells give rise to highly drug resistant cells

<p>Abstract</p> <p>Background</p> <p>Tumor tissue resembles chronically inflamed tissue. Since chronic inflammatory conditions are a strong stimulus for bone marrow-derived cells (BMDCs) it can be assumed that recruitment of BMDCs into cancer tissue should be a common phenomenon. Several data have outlined that BMDC can influence tumor growth and metastasis, e.g., by inducing a paracrine acting feedback loop in tumor cells. Likewise, cell fusion and horizontal gene transfer are further mechanisms how BMDCs can trigger tumor progression.</p> <p>Results</p> <p>Hygromycin resistant murine 67NR-Hyg mammary carcinoma cells were co-cultivated with puromycin resistant murine BMDCs from Tg(GFPU)5Nagy/J mice. Isolation of hygromycin/puromycin resistant mBMDC/67NR-Hyg cell clones was performed by a dual drug selection procedure. PCR analysis revealed an overlap of parental markers in mBMDC/67NR-Hyg cell clones, suggesting that dual resistant cells originated by cell fusion. By contrast, both STR and SNP data analysis indicated that only parental 67NR-Hyg alleles were found in mBMDC/67NR-Hyg cell clones favoring horizontal gene transfer as the mode of origin. RealTime-PCR-array analysis showed a marked up-regulation of Abcb1a and Abcb1b ABC multidrug transporters in mBMDC/67NR-Hyg clones, which was verified by Western Blot analysis. Moreover, the markedly increased Abcb1a/Abcb1b expression was correlated to an efficient Rhodamine 123 efflux, which was completely inhibited by verapamil, a well-known Abcb1a/Abcb1b inhibitor. Likewise, mBMDCs/67NR-Hyg clones revealed a marked resistance towards chemotherapeutic drugs including 17-DMAG, doxorubicin, etoposide and paclitaxel. In accordance to Rhodamine 123 efflux data, chemotherapeutic drug resistance of mBMDC/67NR-Hyg cells was impaired by verapamil mediated blockage of Abc1a/Abcb1b multidrug transporter function.</p> <p>Conclusion</p> <p>Co-cultivation of mBMDCs and mouse 67NR-Hyg mammary carcinoma cells gave rise to highly drug resistant cells. Even though it remains unknown whether mBMDC/67NR-Hyg clones originated by cell fusion or horizontal gene transfer, our data indicate that the exchange of genetic information between two cellular entities is crucial for the origin of highly drug resistant cancer (hybrid) cells, which might be capable to survive chemotherapy.</p

Evaluation of Four-Factor Prothrombin Complex Concentrate Dosing Strategies for the Reversal of Bleeding Associated with Apixaban or Rivaroxaban

In clinical practice, 4F-PCC is commonly used off-label as a non-specific reversal agent for factor Xa inhibitors apixaban and rivaroxaban. While the hemostatic effectiveness of 4F-PCC in patients taking apixaban and rivaroxaban has been studied in the literature, the optimal dose remains unclear. This was a multicenter, retrospective observational study designed to evaluate the hemostatic effectiveness and safety of 4F-PCC dosed at 2000 units, 35 units/kg, and 50 units/kg for the reversal of bleeding associated with apixaban or rivaroxaban. The primary outcome was hemostatic effectiveness as defined by the Internal Society of Thrombosis and Hemostasis and secondary outcomes were rates of all-cause in-hospital mortality and thrombosis at 30 days or discharge. Out of 278 patients who received 4F-PCC for reversal of bleeding associated with apixaban or rivaroxaban, 72 patients were included in the final analysis. The 2000-unit, 35 unit/kg, and 50 unit/kg dosing strategies were used in 12, 36, and 24 patients, respectively. Hemostatic effectiveness was achieved in 86%, 67%, and 70% of intracerebral hemorrhages (p = 0.762) and 60%, 71%, 86% of gastrointestinal hemorrhages (p = 0.422). Neither dosing strategy was associated with a statistically significantly higher rate of hemostatic effectiveness nor lower rates of all-cause mortality or thrombosis

Transient spleen enlargement in peripheral blood progenitor cell donors given G-CSF

The administration of granulocyte colony-stimulating factor (G-CSF) to peripheral blood progenitor cell (PBPC) donors causes spleen length to increase, but the duration of enlargement is not known. Eighteen healthy subjects were given 10 μg/kg of G-CSF for 5 days and a PBSC concentrate was collected by apheresis. Ultrasound scans were used to assess craniocaudal spleen length before and after G-CSF administration. Mean spleen length increased from a baseline length of 10.7 ± 1.3 cm to 12.1 ± 1.2 cm on the apheresis day (p < 0.001). Ten days after apheresis, spleen length fell to 10.5 ± 1.2 cm and did not differ from baseline levels (p = 0.57), but in 3 subjects remained 0.5 cm greater than baseline length. Increases in spleen length in PBPC donors are transient and reversible

Gluon-induced W-boson pair production at the LHC

Pair production of W bosons constitutes an important background to Higgs boson and new physics searches at the Large Hadron Collider LHC. We have calculated the loop-induced gluon-fusion process gg -> W*W* -> leptons, including intermediate light and heavy quarks and allowing for arbitrary invariant masses of the W bosons. While formally of next-to-next-to-leading order, the gg -> W*W* -> leptons process is enhanced by the large gluon flux at the LHC and by experimental Higgs search cuts, and increases the next-to-leading order WW background estimate for Higgs searches by about 30%. We have extended our previous calculation to include the contribution from the intermediate top-bottom massive quark loop and the Higgs signal process. We provide updated results for cross sections and differential distributions and study the interference between the different gluon scattering contributions. We describe important analytical and numerical aspects of our calculation and present the public GG2WW event generator.Comment: 20 pages, 4 figure

Iris: an Extensible Application for Building and Analyzing Spectral Energy Distributions

Iris is an extensible application that provides astronomers with a user-friendly interface capable of ingesting broad-band data from many different sources in order to build, explore, and model spectral energy distributions (SEDs). Iris takes advantage of the standards defined by the International Virtual Observatory Alliance, but hides the technicalities of such standards by implementing different layers of abstraction on top of them. Such intermediate layers provide hooks that users and developers can exploit in order to extend the capabilities provided by Iris. For instance, custom Python models can be combined in arbitrary ways with the Iris built-in models or with other custom functions. As such, Iris offers a platform for the development and integration of SED data, services, and applications, either from the user's system or from the web. In this paper we describe the built-in features provided by Iris for building and analyzing SEDs. We also explore in some detail the Iris framework and software development kit, showing how astronomers and software developers can plug their code into an integrated SED analysis environment.Comment: 18 pages, 8 figures, accepted for publication in Astronomy & Computin

Altered in-stent hemodynamics may cause erroneous upgrading of moderate carotid artery restenosis when evaluated by duplex ultrasound

ObjectiveTo assess the influence of stent application on in-stent hemodynamics under standardized conditions.MethodsOvine common carotid arteries before and after stent (6 × 40 mm, sinus-Carotid-RXt, combined open-closed cell design; Optimed, Ettlingen, Germany) application were used. Plastic tubes, 10 mm in length, simulating stenosis were placed in the middle of the applied stent to induce different degrees of stenosis (moderate 57.8% and severe 76.4%). Flow velocity and dynamic compliance were, respectively, measured with ultrasound and laser scan; proximal, in-stent, and distal to the stented arterial segment (1 cm proximal and distal) in a pulsatile ex vivo circulation system.ResultsStent insertion caused the in-stent peak systolic velocity to increase 22% without stenosis, 31% with moderate stenosis, and 23% with severe stenosis. Stent insertion without stenosis caused no significant increase in in-stent end-diastolic velocity (EDV) but a 17% increase with moderate stenosis. In severe stenosis, EDV was increased 56% proximal to the stenosis. Compliance was reduced threefold in the middle of the stented arterial segment where flow velocity was significantly increased.ConclusionsWith or without stenosis, stent introduction caused the in-stent peak systolic velocity to become significantly elevated compared with a nonstented area. EDV was also increased by stent insertion in the case of moderate stenosis. The stent-induced compliance reduction may be causal for the increase in flow velocity since the stent-induced flow velocity elevation appeared in the stented area with low compliance. Because of altered hemodynamics caused by stent introduction when measured by duplex ultrasound, caution is prudent in concluding that carotid artery stenting is associated with a higher restenosis rate than carotid endarterectomy. Mistakenly upgrading moderate to severe restenosis could result in unnecessary reintervention.Clinical RelevanceClinical experience and prior studies support the supposition that restenosis after carotid artery stenting in carotid lesions displays erroneously elevated velocity when evaluated by duplex ultrasound (DUS), thus contributing to misleading interpretation of the degree of stenosis. This study, in contrast to studies of other groups, employs exactly the same conditions to measure flow with DUS in an unstented and then stented section of the carotid artery. Since DUS is the first-choice tool for carotid artery evaluation, knowledge about inexactness of the method is essential to avoid errors in treatment or follow-up decisions

Mixed reality for the assessment of aortoiliac anatomy in patients with abdominal aortic aneurysm prior to open and endovascular repair: Feasibility and interobserver agreement

Objectives The objective is to evaluate the feasibility and interobserver agreement of a Mixed Reality Viewer (MRV) in the assessment of aortoiliac vascular anatomy of abdominal aortic aneurysm (AAA) patients. Methods Fifty preoperative computed tomography angiographies (CTAs) of AAA patients were included. CTAs were assessed in a mixed reality (MR) environment with respect to aortoiliac anatomy according to a standardized protocol by two experienced observers (Mixed Reality Viewer, MRV, Brainlab AG, Germany). Additionally, all CTAs were independently assessed applying the same protocol by the same observers using a conventional DICOM viewer on a two-dimensional screen with multi-planar reconstructions (Conventional viewer, CV, GE Centricity PACS RA1000 Workstation, GE, United States). The protocol included four sets of items: calcification, dilatation, patency, and tortuosity as well as the number of lumbar and renal arteries. Interobserver agreement (IA, Cohen’s Kappa, κ) was calculated for every item set. Results All CTAs could successfully be displayed in the MRV (100%). The MRV demonstrated equal or better IA in the assessment of anterior and posterior calcification (κMRV: 0.68 and 0.61, κCV: 0.33 and 0.45, respectively) as well as tortuosity (κMRV: 0.60, κCV: 0.48) and dilatation (κMRV: 0.68, κCV: 0.67). The CV demonstrated better IA in the assessment of patency (κMRV: 0.74, κCV: 0.93). The CV also identified significantly more lumbar arteries (CV: 379, MRV: 239, p < 0.01). Conclusions The MRV is a feasible imaging viewing technology in clinical routine. Future efforts should aim at improving hologram quality and enabling accurate registration of the hologram with the physical patient