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1.
FASEB J ; 37(8): e23077, 2023 08.
Article in English | MEDLINE | ID: mdl-37402128

ABSTRACT

Inflammatory processes are activated following ischemic stroke that lead to increased tissue damage for weeks following the ischemic insult, but there are no approved therapies that target this inflammation-induced secondary injury. Here, we report that SynB1-ELP-p50i, a novel protein inhibitor of the nuclear factor kappa B (NF-κB) inflammatory cascade bound to the drug carrier elastin-like polypeptide (ELP), decreases NF-κB induced inflammatory cytokine production in cultured macrophages, crosses the plasma membrane and accumulates in the cytoplasm of both neurons and microglia in vitro, and accumulates at the infarct site where the blood-brain barrier (BBB) is compromised following middle cerebral artery occlusion (MCAO) in rats. Additionally, SynB1-ELP-p50i treatment reduces infarct volume by 11.86% compared to saline-treated controls 24 h following MCAO. Longitudinally, SynB1-ELP-p50i treatment improves survival for 14 days following stroke with no effects of toxicity or peripheral organ dysfunction. These results show high potential for ELP-delivered biologics for therapy of ischemic stroke and other central nervous system disorders and further support targeting inflammation in ischemic stroke.


Subject(s)
Brain Ischemia , Ischemic Stroke , Stroke , Rats , Animals , NF-kappa B/metabolism , Ischemic Stroke/metabolism , Elastin/metabolism , Brain/metabolism , Peptides/pharmacology , Peptides/metabolism , Stroke/metabolism , Brain Ischemia/drug therapy , Brain Ischemia/metabolism , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Anti-Inflammatory Agents/metabolism , Infarction, Middle Cerebral Artery/metabolism , Inflammation/metabolism , Microglia/metabolism
2.
bioRxiv ; 2023 Mar 15.
Article in English | MEDLINE | ID: mdl-36993686

ABSTRACT

Inflammatory processes are activated following ischemic strokes and lead to increased tissue damage for weeks following the ischemic insult, but there are no approved therapies that target this inflammation-induced secondary injury. Here, we report that SynB1-ELP-p50i, a novel protein inhibitor of the nuclear factor kappa B (NF-κB) inflammatory cascade bound to drug carrier elastin-like polypeptide (ELP), is able to enter both neurons and microglia, cross the blood-brain barrier, localize exclusively in the ischemic core and penumbra in Wistar-Kyoto and spontaneously hypertensive rats (SHRs), and reduce infarct volume in male SHRs. Additionally, in male SHRs, SynB1-ELP-p50i treatment improves survival for 14 days following stroke with no effects of toxicity or peripheral organ dysfunction. These results show high potential for ELP-delivered biologics for therapy of ischemic stroke and other central nervous system disorders and further support targeting inflammation in ischemic stroke.

3.
Appl Plant Sci ; 11(1): e11509, 2023.
Article in English | MEDLINE | ID: mdl-36818780

ABSTRACT

Premise: We developed a novel, cost-effective protocol that facilitates testing anoxia tolerance in plants without access to specialized equipment. Methods and Results: Arabidopsis thaliana and barley (Hordeum vulgare) seedlings were treated in airtight 2-L Kilner jars. An anoxic atmosphere was generated using Oxoid AnaeroGen 2.5-L sachets placed on in-house, custom-built wire stands. The performed experiments confirmed a higher sensitivity to low oxygen stress previously observed in anac017 A. thaliana mutants and the positive effect of exogenous sucrose on anoxia tolerance reported by previous studies in A. thaliana. Barley seedlings displayed typical responses to anoxia treatment, including shoot growth cessation and the induction of marker genes for anaerobic metabolism and ethylene biosynthesis in root tissue. Conclusions: The results validate the novel method as an inexpensive, simple alternative for testing anoxia tolerance in plants, where access to an anaerobic workstation is not possible. The novel protocol requires minimum investment and is easily adaptable.

4.
Eur Radiol ; 32(10): 6891-6899, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35567604

ABSTRACT

OBJECTIVES: Successful lung cancer screening delivery requires sensitive, timely reporting of low-dose computed tomography (LDCT) scans, placing a demand on radiology resources. Trained non-radiologist readers and computer-assisted detection (CADe) software may offer strategies to optimise the use of radiology resources without loss of sensitivity. This report examines the accuracy of trained reporting radiographers using CADe support to report LDCT scans performed as part of the Lung Screen Uptake Trial (LSUT). METHODS: In this observational cohort study, two radiographers independently read all LDCT performed within LSUT and reported on the presence of clinically significant nodules and common incidental findings (IFs), including recommendations for management. Reports were compared against a 'reference standard' (RS) derived from nodules identified by study radiologists without CADe, plus consensus radiologist review of any additional nodules identified by the radiographers. RESULTS: A total of 716 scans were included, 158 of which had one or more clinically significant pulmonary nodules as per our RS. Radiographer sensitivity against the RS was 68-73.7%, with specificity of 92.1-92.7%. Sensitivity for detection of proven cancers diagnosed from the baseline scan was 83.3-100%. The spectrum of IFs exceeded what could reasonably be covered in radiographer training. CONCLUSION: Our findings highlight the complexity of LDCT reporting requirements, including the limitations of CADe and the breadth of IFs. We are unable to recommend CADe-supported radiographers as a sole reader of LDCT scans, but propose potential avenues for further research including initial triage of abnormal LDCT or reporting of follow-up surveillance scans. KEY POINTS: • Successful roll-out of mass screening programmes for lung cancer depends on timely, accurate CT scan reporting, placing a demand on existing radiology resources. • This observational cohort study examines the accuracy of trained radiographers using computer-assisted detection (CADe) software to report lung cancer screening CT scans, as a potential means of supporting reporting workflows in LCS programmes. • CADe-supported radiographers were less sensitive than radiologists at identifying clinically significant pulmonary nodules, but had a low false-positive rate and good sensitivity for detection of confirmed cancers.


Subject(s)
Lung Neoplasms , Multiple Pulmonary Nodules , Computers , Early Detection of Cancer/methods , Humans , Lung Neoplasms/diagnostic imaging , Multiple Pulmonary Nodules/diagnostic imaging , Sensitivity and Specificity , Tomography, X-Ray Computed/methods
5.
BMC Gastroenterol ; 21(1): 383, 2021 Oct 18.
Article in English | MEDLINE | ID: mdl-34663234

ABSTRACT

BACKGROUND: Advanced colorectal neoplasms (ACNs), including colorectal cancers (CRC) and high-risk adenomas (HRA), are detected in less than 20% of persons aged 50 years or older who undergo colonoscopy. We sought to derive personalized predictive models of risk of harbouring ACNs to improve colonoscopy wait times for high-risk patients and allocation of colonoscopy resources. METHODS: We characterized colonoscopy indications, neoplasia risk factors and colonoscopy findings through chart review for consecutive individuals aged 50 years or older who underwent outpatient colonoscopy at The Ottawa Hospital (Ottawa, Canada) between April 1, 2008 and March 31, 2012 for non-life threatening indications. We linked patients to population-level health administrative datasets to ascertain additional historical predictor variables and derive multivariable logistic regression models for risk of harboring ACNs at colonoscopy. We assessed model discriminatory capacity and calibration and the ability of the models to improve colonoscopy specificity while maintaining excellent sensitivity for ACN capture. RESULTS: We modelled 17 candidate predictors in 11,724 individuals who met eligibility criteria. The final CRC model comprised 8 variables and had a c-statistic value of 0.957 and a goodness-of-fit p-value of 0.527. Application of the models to our cohort permitted 100% sensitivity for identifying persons with CRC and > 90% sensitivity for identifying persons with HRA, while improving colonoscopy specificity for ACNs by 23.8%. CONCLUSIONS: Our multivariable models show excellent discriminatory capacity for persons with ACNs and could significantly increase colonoscopy specificity without overly sacrificing sensitivity. If validated, these models could allow more efficient allocation of colonoscopy resources, potentially reducing wait times for those at higher risk while deferring unnecessary colonoscopies in low-risk individuals.


Subject(s)
Adenoma , Colorectal Neoplasms , Adenoma/diagnosis , Colonoscopy , Colorectal Neoplasms/diagnosis , Humans , Logistic Models , Risk Factors
6.
Sci Rep ; 11(1): 6216, 2021 03 18.
Article in English | MEDLINE | ID: mdl-33737643

ABSTRACT

Vascular Endothelial Growth Factor (VEGF), a key mediator of angiogenesis and vascular repair, is reduced in chronic ischemic renal diseases, leading to microvascular rarefaction and deterioration of renal function. We developed a chimeric fusion of human VEGF-A121 with the carrier protein Elastin-like Polypeptide (ELP-VEGF) to induce therapeutic angiogenesis via targeted renal VEGF therapy. We previously showed that ELP-VEGF improves renal vascular density, renal fibrosis, and renal function in swine models of chronic renal diseases. However, VEGF is a potent cytokine that induces angiogenesis and increases vascular permeability, which could cause undesired off-target effects or be deleterious in a patient with a solid tumor. Therefore, the current study aims to define the toxicological profile of ELP-VEGF and assess its risk for exacerbating tumor progression and vascularity using rodent models. A dose escalating toxicology assessment of ELP-VEGF was performed by administering a bolus intravenous injection at doses ranging from 0.1 to 200 mg/kg in Sprague Dawley (SD) rats. Blood pressure, body weight, and glomerular filtration rate (GFR) were quantified longitudinally, and terminal blood sampling and renal vascular density measurements were made 14 days after treatment. Additionally, the effects of a single administration of ELP-VEGF (0.1-10 mg/kg) on tumor growth rate, mass, and vascular density were examined in a mouse model of breast cancer. At doses up to 200 mg/kg, ELP-VEGF had no effect on body weight, caused no changes in plasma or urinary markers of renal injury, and did not induce renal fibrosis or other histopathological findings in SD rats. At the highest doses (100-200 mg/kg), ELP-VEGF caused an acute, transient hypotension (30 min), increased GFR, and reduced renal microvascular density 14 days after injection. In a mouse tumor model, ELP-VEGF did not affect tumor growth rate or tumor mass, but analysis of tumor vascular density by micro-computed tomography (µCT) revealed significant, dose dependent increases in tumor vascularity after ELP-VEGF administration. ELP-VEGF did not induce toxicity in the therapeutic dosing range, and doses one hundred times higher than the expected maximum therapeutic dose were needed to observe any adverse signs in rats. In breast tumor-bearing mice, ELP-VEGF therapy induced a dose-dependent increase in tumor vascularity, demanding caution for potential use in a patient suffering from kidney disease but with known or suspected malignancy.


Subject(s)
Biological Products/pharmacology , Breast Neoplasms/blood supply , Elastin/genetics , Neovascularization, Pathologic/chemically induced , Recombinant Fusion Proteins/pharmacology , Renal Insufficiency, Chronic/drug therapy , Vascular Endothelial Growth Factor A/genetics , Animals , Biological Products/metabolism , Blood Pressure/drug effects , Body Weight/drug effects , Breast Neoplasms/diagnostic imaging , Breast Neoplasms/pathology , Capillary Permeability/drug effects , Disease Models, Animal , Elastin/metabolism , Female , Gene Expression , Glomerular Filtration Rate/drug effects , Heterografts , Humans , Hypotension/chemically induced , Hypotension/diagnostic imaging , Hypotension/physiopathology , Mice , Molecular Mimicry , Neovascularization, Pathologic/diagnostic imaging , Neovascularization, Pathologic/pathology , Neovascularization, Physiologic/drug effects , Rats , Rats, Sprague-Dawley , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Renal Insufficiency, Chronic/diagnostic imaging , Renal Insufficiency, Chronic/metabolism , Renal Insufficiency, Chronic/physiopathology , Swine , Toxicity Tests, Chronic , Vascular Endothelial Growth Factor A/metabolism , X-Ray Microtomography
7.
Thorax ; 75(10): 908-912, 2020 10.
Article in English | MEDLINE | ID: mdl-32759387

ABSTRACT

The Lung Screen Uptake Trial tested a novel invitation strategy to improve uptake and reduce socioeconomic and smoking-related inequalities in lung cancer screening (LCS) participation. It provides one of the first UK-based 'real-world' LCS cohorts. Of 2012 invited, 1058 (52.6%) attended a 'lung health check'. 768/996 (77.1%) in the present analysis underwent a low-dose CT scan. 92 (11.9%) and 33 (4.3%) participants had indeterminate pulmonary nodules requiring 3-month and 12-month surveillance, respectively; 36 lung cancers (4.7%) were diagnosed (median follow-up: 1044 days). 72.2% of lung cancers were stage I/II and 79.4% of non-small cell lung cancer had curative-intent treatment.


Subject(s)
Carcinoma/diagnosis , Early Detection of Cancer , Lung Neoplasms/diagnosis , Patient Acceptance of Health Care , Aged , Cohort Studies , Female , Humans , Male , Middle Aged , Neoplasm Staging , Radiation Dosage , Socioeconomic Factors , United Kingdom
8.
Emerg Infect Dis ; 26(8)2020 Aug.
Article in English | MEDLINE | ID: mdl-32396505

ABSTRACT

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was identified as the etiologic agent associated with coronavirus disease, which emerged in late 2019. In response, we developed a diagnostic panel consisting of 3 real-time reverse transcription PCR assays targeting the nucleocapsid gene and evaluated use of these assays for detecting SARS-CoV-2 infection. All assays demonstrated a linear dynamic range of 8 orders of magnitude and an analytical limit of detection of 5 copies/reaction of quantified RNA transcripts and 1 x 10-1.5 50% tissue culture infectious dose/mL of cell-cultured SARS-CoV-2. All assays performed comparably with nasopharyngeal and oropharyngeal secretions, serum, and fecal specimens spiked with cultured virus. We obtained no false-positive amplifications with other human coronaviruses or common respiratory pathogens. Results from all 3 assays were highly correlated during clinical specimen testing. On February 4, 2020, the Food and Drug Administration issued an Emergency Use Authorization to enable emergency use of this panel.


Subject(s)
Betacoronavirus/genetics , Coronavirus Infections/diagnosis , Nucleocapsid Proteins/genetics , Pneumonia, Viral/diagnosis , RNA, Viral/genetics , Real-Time Polymerase Chain Reaction/methods , Biomarkers/analysis , COVID-19 , Centers for Disease Control and Prevention, U.S. , Coronavirus Infections/virology , Coronavirus Nucleocapsid Proteins , DNA Primers/chemical synthesis , DNA Primers/genetics , Feces/virology , Fluoresceins/chemistry , Fluorescent Dyes/chemistry , Humans , Limit of Detection , Nasopharynx/virology , Pandemics , Phosphoproteins , Pneumonia, Viral/virology , Real-Time Polymerase Chain Reaction/standards , Reproducibility of Results , SARS-CoV-2 , Sputum/virology , United States
9.
Ann Am Thorac Soc ; 17(7): 869-878, 2020 07.
Article in English | MEDLINE | ID: mdl-32164439

ABSTRACT

Rationale: Individuals eligible for lung cancer screening (LCS) by low-dose computed tomography (LDCT) are also at risk of chronic obstructive pulmonary disease (COPD) due to age and smoking exposure. Whether the LCS episode is useful for early detection of COPD is not well established.Objectives: To explore associations between symptoms, comorbidities, spirometry, and emphysema in participants enrolled in the Lung Screen Uptake Trial.Methods: This cross-sectional study was a prespecified analysis nested within Lung Screen Uptake Trial, which was a randomized study testing the impact of differing invitation materials on attendance of 60- to 75-year-old smokers and ex-smokers to a "lung health check" between November 2015 and July 2017. Participants with a smoking history ≥30 pack-years and who quit ≤15 years ago, or meeting a lung cancer risk of ≥1.51% via the Prostate Lung Colorectal Ovarian model or ≥2.5% via the Liverpool Lung Project model, were offered LDCT. COPD was defined and classified according to the GOLD (Global Initiative for Obstructive Lung Disease) criteria using prebronchodilator spirometry. Analyses included the use of descriptive statistics, chi-square tests to examine group differences, and univariable and multivariable logistic regression to explore associations between symptom prevalence, airflow limitation, and visually graded emphysema.Results: A total of 560 of 986 individuals included in the analysis (57%) had prebronchodilator spirometry consistent with COPD; 67% did not have a prior history of COPD and were termed "undiagnosed." Emphysema prevalence in those with known and "undiagnosed" COPD was 73% and 68%, respectively. A total of 32% of those with "undiagnosed COPD" had no emphysema on LDCT. Inhaler use and symptoms were more common in the "known" than the "undiagnosed" COPD group (63% vs. 33% with persistent cough [P < 0.001]; 73% vs. 33% with dyspnea [P < 0.001]). Comorbidities were common in all groups. Adjusted odds ratio (aOR) of respiratory symptoms were more significant for airflow obstruction (aOR GOLD 1 and 2, 1.57; confidence interval [CI], 1.14-2.17; aOR GOLD 3 and 4, 4.6; CI, 2.17-9.77) than emphysema (aOR mild, 1.12; CI, 0.81-1.55; aOR moderate, 1.33; CI, 0.85-2.09; aOR severe, 4.00; CI, 1.57-10.2).Conclusions: There is high burden of "undiagnosed COPD" and emphysema in LCS participants. Adding spirometry findings to the LDCT enhances identification of individuals with COPD.Clinical trial registered with www.clinicaltrials.gov (NCT02558101).


Subject(s)
Lung Neoplasms/diagnostic imaging , Mass Screening/methods , Pulmonary Disease, Chronic Obstructive/diagnosis , Smoking/adverse effects , Aged , Cough/etiology , Cross-Sectional Studies , Early Detection of Cancer , Emphysema/diagnostic imaging , Female , Humans , Logistic Models , Male , Middle Aged , Multivariate Analysis , Prevalence , Pulmonary Disease, Chronic Obstructive/epidemiology , Spirometry , Tomography, X-Ray Computed , United Kingdom/epidemiology
10.
Thorax ; 74(12): 1140-1146, 2019 12.
Article in English | MEDLINE | ID: mdl-31558626

ABSTRACT

INTRODUCTION: Lung cancer screening (LCS) by low-dose computed tomography (LDCT) offers an opportunity to impact both lung cancer and coronary heart disease mortality through detection of coronary artery calcification (CAC). Here, we explore the value of CAC and cardiovascular disease (CVD) risk assessment in LCS participants in the Lung Screen Uptake Trial (LSUT). METHODS: In this cross-sectional study, current and ex-smokers aged 60-75 were invited to a 'lung health check'. Data collection included a CVD risk assessment enabling estimation of 10 year CVD risk using the QRISK2 score. Participants meeting the required lung cancer risk underwent an ungated, non-contrast LDCT. Descriptive data, bivariate associations and a multivariate analysis of predictors of statin use are presented. RESULTS: Of 1005 individuals enrolled, 680 were included in the final analysis. 421 (61.9%) had CAC present and in 49 (7.2%), this was heavy. 668 (98%) of participants had a QRISK2≥10% and QRISK2 was positively associated with increasing CAC grade (OR 4.29 (CI 0.93 to 19.88) for QRISK2=10%-20% and 12.29 (CI 2.68 to 56.1) for QRISK2≥20% respectively). Of those who qualified for statin primary prevention (QRISK2≥10%), 56.8% did not report a history of statin use. In the multivariate analysis statin use was associated with age, body mass index and history of hypertension and diabetes. CONCLUSIONS: LCS offers an important opportunity for instituting CVD risk assessment in all LCS participants irrespective of the presence of LDCT-detected CAC. Further studies are needed to determine whether CAC could enhance uptake and adherence to primary preventative strategies.


Subject(s)
Cardiovascular Diseases/prevention & control , Early Detection of Cancer/methods , Lung Neoplasms/diagnostic imaging , Aged , Cardiovascular Diseases/complications , Cohort Studies , Coronary Disease/complications , Coronary Disease/diagnostic imaging , Cross-Sectional Studies , Drug Utilization/statistics & numerical data , Female , Humans , Hydroxymethylglutaryl-CoA Reductase Inhibitors/therapeutic use , Lung Neoplasms/complications , Male , Mass Screening/methods , Middle Aged , Primary Prevention/methods , Prospective Studies , Radiation Dosage , Risk Assessment/methods , Tomography, X-Ray Computed/methods , Vascular Calcification/complications , Vascular Calcification/diagnostic imaging
12.
Acad Radiol ; 25(12): 1556-1563, 2018 12.
Article in English | MEDLINE | ID: mdl-29724674

ABSTRACT

RATIONALE AND OBJECTIVES: Chest X-rays (CXR) are one of the most frequently requested imaging examinations and are fundamental to many patient pathways. The aim of this study was to investigate the diagnostic accuracy of CXR interpretation by reporting radiographers (technologists). METHODS: A cohort of consultant radiologists (n = 10) and reporting radiographers (technologists; n = 11) interpreted a bank (n = 106) of adult CXRs that contained a range of pathologies. Jack-knife alternate free-response receiver operating characteristic (JAFROC) methodology was used to determine the performance of the observers (JAFROC v4.2). A noninferiority approach was used, with a predefined margin of clinical insignificance of 10% of average consultant radiologist diagnostic accuracy. RESULTS: The diagnostic accuracy of the reporting radiographers (figure of merit = 0.828, 95% confidence interval 0.808-0.847) was noninferior to the consultant radiologists (figure of merit = 0.788, 95% confidence interval 0.766-0.811), P < .0001. CONCLUSIONS: With appropriate postgraduate education, reporting radiographers are able to interpret CXRs at a level comparable to consultant radiologists.


Subject(s)
Allied Health Personnel/standards , Heart Diseases/diagnostic imaging , Lung Diseases/diagnostic imaging , Radiologists/standards , Diagnostic Errors , Humans , Observer Variation , Prospective Studies , ROC Curve , Radiography, Thoracic
13.
J Med Radiat Sci ; 65(3): 200-208, 2018 Sep.
Article in English | MEDLINE | ID: mdl-29806102

ABSTRACT

INTRODUCTION: Diagnostic capacity and time to diagnosis are frequently identified as a barrier to improving cancer patient outcomes. Maximising the contribution of the medical imaging workforce, including reporting radiographers, is one way to improve service delivery. METHODS: An efficient and effective centralised model of workplace training support was designed for a cohort of trainee chest X-ray (CXR) reporting radiographers. A comprehensive schedule of tutorials was planned and aligned with the curriculum of a post-graduate certificate in CXR reporting. Trainees were supported via a hub and spoke model (centralised training model), with the majority of education provided by a core group of experienced CXR reporting radiographers. Trainee and departmental feedback on the model was obtained using an online survey. RESULTS: Fourteen trainees were recruited from eight National Health Service Trusts across London. Significant efficiencies of scale were possible with centralised support (48 h) compared to traditional workplace support (348 h). Trainee and manager feedback overall was positive. Trainees and managers both reported good trainee support, translation of learning to practice and increased confidence. Logistics, including trainee travel and release, were identified as areas for improvement. CONCLUSION: Centralised workplace training support is an effective and efficient method to create sustainable diagnostic capacity and support improvements in the lung cancer pathway.


Subject(s)
Inservice Training/methods , Lung Neoplasms/diagnostic imaging , Radiography, Thoracic/standards , Radiologists/education , Adult , England , Female , Humans , Inservice Training/standards , Male , Radiologists/standards , Teaching Materials
14.
Emerg Infect Dis ; 24(3): 471-477, 2018 03.
Article in English | MEDLINE | ID: mdl-29460739

ABSTRACT

In 2010, the Centers for Disease Control and Prevention began to develop an Influenza Risk Assessment Tool (IRAT) to methodically capture and assess information relating to influenza A viruses not currently circulating among humans. The IRAT uses a multiattribute, additive model to generate a summary risk score for each virus. Although the IRAT is not intended to predict the next pandemic influenza A virus, it has provided input into prepandemic preparedness decisions.


Subject(s)
Influenza A virus , Influenza, Human/epidemiology , Influenza, Human/virology , Animals , China/epidemiology , Genotype , Humans , Influenza A virus/classification , Influenza A virus/genetics , Orthomyxoviridae Infections/epidemiology , Orthomyxoviridae Infections/virology , Population Surveillance , Risk Assessment
15.
J Immunol ; 200(3): 1110-1123, 2018 02 01.
Article in English | MEDLINE | ID: mdl-29263214

ABSTRACT

TNF-related apoptosis-inducing ligand (TRAIL) was initially described to induce apoptosis of tumor cells and/or virally infected cells, although sparing normal cells, and has been implicated in the pathogenesis of HIV disease. We previously identified TRAILshort, a TRAIL splice variant, in HIV-infected patients and characterized it as being a dominant negative ligand to subvert TRAIL-mediated killing. Herein, using single-cell genomics we demonstrate that TRAILshort is produced by HIV-infected cells, as well as by uninfected bystander cells, and that the dominant stimulus which induces TRAILshort production are type I IFNs and TLR7, TLR8, and TLR9 agonists. TRAILshort has a short t1/2 by virtue of containing a PEST domain, which targets the protein toward the ubiquitin proteasome pathway for degradation. Further we show that TRAILshort binds preferentially to TRAIL receptors 1 and 2 with significantly reduced interaction with the decoy TRAIL receptors 3 and 4. Recombinant TRAILshort is sufficient to protect cells against TRAIL-induced killing, whereas immunodepletion of TRAILshort with a specific Ab restores TRAIL sensitivity. Importantly we show that TRAILshort is shed in microvesicles into the cellular microenvironment and therefore confers TRAIL resistance not only on the cell which produces it, but also upon neighboring bystander cells. These results establish a novel paradigm for understanding and overcoming TRAIL resistance, in particular how HIV-infected cells escape immune elimination by the TRAIL:TRAILshort receptor axis.


Subject(s)
CD4-Positive T-Lymphocytes/immunology , Cellular Microenvironment/immunology , HIV Infections/immunology , Protein Isoforms/genetics , Receptors, TNF-Related Apoptosis-Inducing Ligand/metabolism , TNF-Related Apoptosis-Inducing Ligand/genetics , Alternative Splicing/genetics , Apoptosis , Bystander Effect/immunology , CD4-Positive T-Lymphocytes/virology , Cell Line, Tumor , Cell Membrane/immunology , HEK293 Cells , HIV Infections/pathology , HIV Infections/virology , HeLa Cells , Humans , Jurkat Cells , Protein Isoforms/biosynthesis , TNF-Related Apoptosis-Inducing Ligand/biosynthesis
16.
Knee Surg Sports Traumatol Arthrosc ; 26(4): 1252-1257, 2018 Apr.
Article in English | MEDLINE | ID: mdl-28646381

ABSTRACT

PURPOSE AND HYPOTHESIS: The principal purpose of this paper was to identify whether femoral notch morphology was different in females without anterior cruciate ligament (ACL) injury from those with ACL injury. Magnetic resonance imaging (MRI) was used to assess the femoral notch type, notch width index and 'α angle' in female patients and measure these differences. METHODS: This is a retrospective case control study of 119 female patients, 58 with ACL injury and 61 patients without ACL injury who underwent knee MRI between March 2014 and April 2016. The morphometric measurements were taken by two independent observers. The femoral notch width index was calculated as the ratio between the central notch width and transcondylar or intercondylar width; values >0.27 were considered normal. The femoral notch shape was classified as Type A, Type U or Type W, with Type A describing a stenotic notch, Type U a notch with a wider contour and Type W a wider Type U with two apices apparent. The angle between the longitudinal femoral axis and the Blumensaat line was identified as the 'α angle'. The statistical analysis was performed with t tests, simple and multivariable logistic regression analysis to evaluate the strength of these specific femoral notch morphometric values as predictive factors to ACL rupture. RESULTS: Stenotic femoral notch Type A was identified as a high risk factor to ACL injury (odds ratio [OR] = 2.8; p = 0.03). There was no significant difference between the two groups for the notch width index (OR = 0.7; p = n.s.) and the 'α angle' (OR 1.02; p = n.s.). Significant association between NWI and stenotic notch was found (p < 0.01). CONCLUSIONS: This study showed that Type A stenotic femoral notch can be considered as a valuable predictive factor for ACL injury. Notch width index and 'α angle' are weak indicators in ACL injury prognosis. Ligament impingement may be inferred as an important mechanism in female ACL rupture. Injury prevention strategies, such as prehabilitation programmes, could be introduced in the benefit of young females with stenotic notch. LEVEL OF EVIDENCE: III.


Subject(s)
Anterior Cruciate Ligament Injuries/etiology , Constriction, Pathologic/diagnostic imaging , Femur/diagnostic imaging , Adult , Anterior Cruciate Ligament Injuries/diagnostic imaging , Anterior Cruciate Ligament Injuries/surgery , Case-Control Studies , Constriction, Pathologic/complications , Female , Femur/anatomy & histology , Humans , Magnetic Resonance Imaging/methods , Retrospective Studies , Risk Factors , Young Adult
17.
J Virol Methods ; 228: 151-7, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26640122

ABSTRACT

In this study, a multicenter evaluation of the Life Technologies TaqMan(®) Array Card (TAC) with 21 custom viral and bacterial respiratory assays was performed on the Applied Biosystems ViiA™ 7 Real-Time PCR System. The goal of the study was to demonstrate the analytical performance of this platform when compared to identical individual pathogen specific laboratory developed tests (LDTs) designed at the Centers for Disease Control and Prevention (CDC), equivalent LDTs provided by state public health laboratories, or to three different commercial multi-respiratory panels. CDC and Association of Public Health Laboratories (APHL) LDTs had similar analytical sensitivities for viral pathogens, while several of the bacterial pathogen APHL LDTs demonstrated sensitivities one log higher than the corresponding CDC LDT. When compared to CDC LDTs, TAC assays were generally one to two logs less sensitive depending on the site performing the analysis. Finally, TAC assays were generally more sensitive than their counterparts in three different commercial multi-respiratory panels. TAC technology allows users to spot customized assays and design TAC layout, simplify assay setup, conserve specimen, dramatically reduce contamination potential, and as demonstrated in this study, analyze multiple samples in parallel with good reproducibility between instruments and operators.


Subject(s)
Oligonucleotide Array Sequence Analysis/standards , Real-Time Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/standards , Bacteria/genetics , Bacteria/isolation & purification , Centers for Disease Control and Prevention, U.S. , Humans , Microfluidics/methods , Microfluidics/standards , Real-Time Polymerase Chain Reaction/instrumentation , Reproducibility of Results , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/microbiology , Respiratory Tract Infections/virology , Sensitivity and Specificity , United States , Viruses/genetics , Viruses/isolation & purification
18.
Rep U S ; 2015: 216-221, 2015.
Article in English | MEDLINE | ID: mdl-26709364

ABSTRACT

A system for actively changing the stiffness of a long, thin, flexible robotic manipulator has been designed for cardiologists to use in a range of diagnosis and treatment procedures. Low-stiffness manipulators, such as catheters, are ideal for steering through vasculature with low risk of tissue injury. However, such instruments are not well-suited for applying force to tissue. The proposed system solves this problem by using a series of bead-shaped vertebrae containing pull wires to actively change the stiffness of the catheter, similar to gooseneck surgical retractors. Individual wires steer the catheter to a desired location. All wires are then tensioned to create friction between each vertebra and prevent sliding, therefore resisting motion. While this design concept has been implemented manually in various settings for decades, fine robotic control of the friction and stiffness of the system relies on a thorough understanding of the friction properties between vertebral segments. We have developed an analytical model to understand the interactions between vertebrae and determine the relationships between system parameters and the overall stiffness of the catheter. Experiments validated the calculations from the model and the functionality of the system by applying known loads to the tip of the catheter and measuring the catheter displacement. The catheter stiffness was measured to range from 100 N/m to 800 N/m, which is sufficient for performing many surgical tasks on tissue. This system can be useful in minimally invasive procedures involving direct instrument contact with tissue by improving accuracy, safety, and work flow.

19.
Emerg Infect Dis ; 21(8): 1372-8, 2015 Aug.
Article in English | MEDLINE | ID: mdl-26196098

ABSTRACT

Although predicting which influenza virus subtype will cause the next pandemic is not yet possible, public health authorities must continually assess the pandemic risk associated with animal influenza viruses, particularly those that have caused infections in humans, and determine what resources should be dedicated to mitigating that risk. To accomplish this goal, a risk assessment framework was created in collaboration with an international group of influenza experts. Compared with the previously used approach, this framework, named the Influenza Risk Assessment Tool, provides a systematic and transparent approach for assessing and comparing threats posed primarily by avian and swine influenza viruses. This tool will be useful to the international influenza community and will remain flexible and responsive to changing information.


Subject(s)
Influenza, Human/epidemiology , Pandemics/statistics & numerical data , Public Health/methods , Risk Assessment/methods , Humans , Influenza, Human/immunology , Influenza, Human/mortality , Public Health/instrumentation
20.
Soc Work Public Health ; 30(1): 30-7, 2015.
Article in English | MEDLINE | ID: mdl-25317978

ABSTRACT

The social determinants of health represent the societal and economic influences responsible for most health inequities. Advocacy to eliminate health inequities for homeless children oftentimes involves the use of community-based approaches. This article details the Floating Hospital's (TFH) community-based participatory research (CBPR) project that resulted in an advocacy brief. Within the project, the community practice concepts of a strengths perspective, empowerment, capacity building, and advocacy are embedded. The brief enhances TFH's capacity to advocate for the needs of homeless children. This example serves as a guide for social work and public health professionals to use CBPR to address health inequities within their communities.


Subject(s)
Child Advocacy , Community-Based Participatory Research , Health Status Disparities , Homeless Youth , Public Health , Capacity Building , Child , Community-Institutional Relations , Female , Humans , Male , Pennsylvania , Social Determinants of Health
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