Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 25
Filter
1.
Article in English | MEDLINE | ID: mdl-38708143

ABSTRACT

While minimally invasive laparoscopic surgery can help reduce blood loss, reduce hospital time, and shorten recovery time compared to open surgery, it has the disadvantages of limited field of view and difficulty in locating subsurface targets. Our proposed solution applies an augmented reality (AR) system to overlay pre-operative images, such as those from magnetic resonance imaging (MRI), onto the target organ in the user's real-world environment. Our system can provide critical information regarding the location of subsurface lesions to guide surgical procedures in real time. An infrared motion tracking camera system was employed to obtain real-time position data of the patient and surgical instruments. To perform hologram registration, fiducial markers were used to track and map virtual coordinates to the real world. In this study, phantom models of each organ were constructed to test the reliability and accuracy of the AR-guided laparoscopic system. Root mean square error (RMSE) was used to evaluate the targeting accuracy of the laparoscopic interventional procedure. Our results demonstrated a registration error of 2.42 ± 0.79 mm and a procedural targeting error of 4.17 ± 1.63 mm using our AR-guided laparoscopic system that will be further refined for potential clinical procedures.

2.
Nat Commun ; 15(1): 3113, 2024 Apr 10.
Article in English | MEDLINE | ID: mdl-38600097

ABSTRACT

Autophagy is a conserved, catabolic process essential for maintaining cellular homeostasis. Malfunctional autophagy contributes to neurodevelopmental and neurodegenerative diseases. However, the exact role and targets of autophagy in human neurons remain elusive. Here we report a systematic investigation of neuronal autophagy targets through integrated proteomics. Deep proteomic profiling of multiple autophagy-deficient lines of human induced neurons, mouse brains, and brain LC3-interactome reveals roles of neuronal autophagy in targeting proteins of multiple cellular organelles/pathways, including endoplasmic reticulum (ER), mitochondria, endosome, Golgi apparatus, synaptic vesicle (SV) for degradation. By combining phosphoproteomics and functional analysis in human and mouse neurons, we uncovered a function of neuronal autophagy in controlling cAMP-PKA and c-FOS-mediated neuronal activity through selective degradation of the protein kinase A - cAMP-binding regulatory (R)-subunit I (PKA-RI) complex. Lack of AKAP11 causes accumulation of the PKA-RI complex in the soma and neurites, demonstrating a constant clearance of PKA-RI complex through AKAP11-mediated degradation in neurons. Our study thus reveals the landscape of autophagy degradation in human neurons and identifies a physiological function of autophagy in controlling homeostasis of PKA-RI complex and specific PKA activity in neurons.


Subject(s)
Neurons , Proteomics , Mice , Animals , Humans , Neurons/metabolism , Cyclic AMP-Dependent Protein Kinases/metabolism , Autophagy/physiology , Homeostasis
3.
J Am Coll Emerg Physicians Open ; 5(2): e13140, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38567033

ABSTRACT

Objective: Protocols to evaluate for myocardial infarction (MI) using high-sensitivity cardiac troponin (hs-cTn) have the potential to drive costs upward due to the added sensitivity. We performed an economic evaluation of an accelerated protocol (AP) to evaluate for MI using hs-cTn to identify changes in costs of treatment and length of stay compared with conventional testing. Methods: We performed a planned secondary economic analysis of a large, cluster randomized trial across nine emergency departments (EDs) from July 2020 to April 2021. Patients were included if they were 18 years or older with clinical suspicion for MI. In the AP, patients could be discharged without further testing at 0 h if they had a hs-cTnI < 4 ng/L and at 1 h if the initial value were 4 ng/L and the 1-h value ≤7 ng/L. Patients in the standard of care (SC) protocol used conventional cTn testing at 0 and 3 h. The primary outcome was the total cost of treatment, and the secondary outcome was ED length of stay. Results: Among 32,450 included patients, an AP had no significant differences in cost (+$89, CI: -$714, $893 hospital cost, +$362, CI: -$414, $1138 health system cost) or ED length of stay (+46, CI: -28, 120 min) compared with the SC protocol. In lower acuity, free-standing EDs, patients under the AP experienced shorter length of stay (-37 min, CI: -62, 12 min) and reduced health system cost (-$112, CI: -$250, $25). Conclusion: Overall, the implementation of AP using hs-cTn does not result in higher costs.

4.
SLAS Discov ; 28(5): 233-239, 2023 07.
Article in English | MEDLINE | ID: mdl-36990319

ABSTRACT

Galectin-3 is a beta-galactoside-binding mammalian lectin that is one of a 15-member galectin family that can bind several cell surface glycoproteins via its carbohydrate recognition domain (CRD). As a result, it can influence a range of cellular processes including cell activation, adhesion and apoptosis. Galectin-3 has been implicated in various diseases, including fibrotic disorders and cancer, and is now being therapeutically targeted by both small and large molecules. Historically, the screening and triaging of small molecule glycomimetics that bind to the galectin-3 CRD has been completed in fluorescence polarisation (FP) assays to determine KD values. Surface plasmon resonance (SPR) has not been widely used for compound screening and in this study it was used to compare human and mouse galectin-3 affinity measures between FP and SPR, as well as investigate compound kinetics. The KD estimates for a set of compounds selected from mono- and di-saccharides with affinities across a 550-fold range, correlated well between FP and SPR assay formats for both human and mouse galectin-3. Increases in affinity for compounds binding to human galectin-3 were driven by changes in both kon and koff whilst for mouse galectin-3 this was primarily due to kon. The reduction in affinity observed between human to mouse galectin-3 was also comparable between assay formats. SPR has been shown to be a viable alternative to FP for early drug discovery screening and determining KD values. In addition, it can also provide early kinetic characterisation of small molecule galectin-3 glycomimetics with robust kon and koff values generated in a high throughput manner.


Subject(s)
Galectin 3 , Surface Plasmon Resonance , Humans , Animals , Mice , Galectin 3/genetics , Galectin 3/chemistry , Galectin 3/metabolism , Kinetics , Galectins/chemistry , Galectins/metabolism , Carbohydrates/chemistry , Mammals/metabolism
5.
BMJ Case Rep ; 14(10)2021 Oct 19.
Article in English | MEDLINE | ID: mdl-34667036

ABSTRACT

Previous reports have described non-ischaemic cardiomyopathy related to a variety of autoimmune diseases. However, very few case reports describe Sjögren disease as a contributing factor to cardiomyopathy. We report the case of a 69-year-old woman with a history of Sjögren disease who presented with cardiogenic shock. Laboratory testing and cardiac MRI revealing apical septal late gadolinium enhancement were consistent with an autoimmune aetiology. After ruling out ischaemic, infectious and other possible causes, the patient's clinical presentation was thought to be related to underlying Sjögren disease. She was treated with intravenous steroids and evidence-based heart failure therapy, but she eventually died after having declined heart transplantation. Given the rarity of Sjögren disease, no diagnostic criteria or standard treatment has been established for cardiomyopathy related to this disease. Diagnosis should be considered in patients who show evidence of autoimmune processes after other possible causes are ruled out.


Subject(s)
Cardiomyopathies , Sjogren's Syndrome , Aged , Cardiomyopathies/complications , Cardiomyopathies/diagnosis , Contrast Media , Female , Gadolinium , Humans , Shock, Cardiogenic/diagnosis , Shock, Cardiogenic/etiology , Sjogren's Syndrome/complications , Sjogren's Syndrome/diagnosis , Sjogren's Syndrome/drug therapy
6.
J Ultrasound Med ; 40(1): 175-181, 2021 Jan.
Article in English | MEDLINE | ID: mdl-32557791

ABSTRACT

When evaluating patients with hip pain, clinicians may be trained to both evaluate for a hip effusion and perform ultrasound-guided arthrocentesis to evaluate the etiology of the effusion. We present a novel 3-dimensional-printed hip arthrocentesis model, which can be used to train clinicians to perform both tasks under ultrasound guidance. Our model uses a combination of a 3-dimensional-printed hip joint, as well as readily available materials such as an infant Ambu (Ballerup, Denmark) bag, syringe, intravenous line kit, and silicone. We present our experience so that others may use and adapt our model for their training purposes.


Subject(s)
Arthrocentesis , Arthralgia , Hip Joint/diagnostic imaging , Hip Joint/surgery , Humans , Ultrasonography , Ultrasonography, Interventional
7.
J Invasive Cardiol ; 32(3): 104-109, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31941835

ABSTRACT

BACKGROUND: Recent studies suggest that primary percutaneous coronary intervention (PCI) and targeted temperature management (TTM) improve outcome in ST-segment elevation myocardial infarction (STEMI) complicated by out-of-hospital cardiac arrest (OHCA). The objective of this study was to evaluate a contemporary series of patients with STEMI and OHCA to characterize treatment approaches and predictors of neurologic outcome. METHODS: From January 2009 through November 2012, a total of 239 patients who underwent emergent coronary angiography at 10 medical centers across the United States were enrolled. All patients suffered OHCA with STEMI on either the prehospital or post-resuscitation electrocardiogram. Neurologic outcome was assessed using the cerebral performance category (CPC) score. Predictors of neurologic outcome were determined using multivariate logistic regression analysis. The primary endpoint was in-hospital survival with good neurologic function (CPC score 1 or 2). RESULTS: Mean age was 60 ± 13 years, 72% were male, and the majority of patients had a history of cardiovascular event. Initial rhythm was ventricular fibrillation in 72%. At hospital presentation, 76% of patients were intubated, 37% were in cardiogenic shock, and 33% were receiving vasopressors. Primary PCI was performed in 74%, with an average door-to-balloon time of 95 ± 77 minutes, and TTM was used in 51%. Forty-four percent of patients had full neurologic recovery (CPC score 1) and 55% had good neurologic function. Overall in-hospital survival rate was 66%. Independent predictors of in-hospital survival with good neurologic function were: receiving bystander cardiopulmonary resuscitation, location of arrest, receiving drug-eluting stents, and not experiencing a recurrent cardiac arrest. CONCLUSIONS: Short-term survival for patients with STEMI and OHCA undergoing emergent coronary angiography and revascularization with TTM in this contemporary, multicenter registry was high and neurologic outcome was good in more than half of patients.


Subject(s)
Myocardial Infarction , Out-of-Hospital Cardiac Arrest , Percutaneous Coronary Intervention , ST Elevation Myocardial Infarction , Aged , Female , Humans , Male , Middle Aged , Out-of-Hospital Cardiac Arrest/diagnosis , Out-of-Hospital Cardiac Arrest/therapy , Registries , ST Elevation Myocardial Infarction/diagnosis , ST Elevation Myocardial Infarction/surgery , Treatment Outcome
8.
Sci Rep ; 9(1): 14645, 2019 10 10.
Article in English | MEDLINE | ID: mdl-31601934

ABSTRACT

Multidomain proteins represent a broad spectrum of the protein landscape and are involved in various interactions. They could be considered as modular building blocks assembled in distinct fashion and connected by linkers of varying lengths and sequences. Due to their intrinsic flexibility, these linkers provide proteins a subtle way to modulate interactions and explore a wide range of conformational space. In the present study, we are seeking to understand the effect of the flexibility and dynamics of the linker involved in the STAM2 UIM-SH3 dual domain protein with respect to molecular recognition. We have engineered several constructs of UIM-SH3 with different length linkers or domain deletion. By means of SAXS and NMR experiments, we have shown that the modification of the linker modifies the flexibility and the dynamics of UIM-SH3. Indeed, the global tumbling of both the UIM and SH3 domain is different but not independent from each other while the length of the linker has an impact on the ps-ns time scale dynamics of the respective domains. Finally, the modification of the flexibility and dynamics of the linker has a drastic effect on the interaction of UIM-SH3 with Lys63-linked diubiquitin with a roughly eight-time weaker dissociation constant.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Endosomal Sorting Complexes Required for Transport/metabolism , Ubiquitins/metabolism , src Homology Domains , Endosomal Sorting Complexes Required for Transport/ultrastructure , Molecular Dynamics Simulation , Nuclear Magnetic Resonance, Biomolecular , Protein Binding , Scattering, Small Angle , Ubiquitins/ultrastructure , X-Ray Diffraction
9.
Cardiovasc Revasc Med ; 20(2): 133-136, 2019 02.
Article in English | MEDLINE | ID: mdl-29898868

ABSTRACT

INTRODUCTION: The purpose of the study is to develop an optimal TR-Band weaning strategy while minimizing vascular access site complications of hematoma or radial artery occlusion (RAO). METHODS: The trial was a randomized, prospective, single center study of 129 patients who underwent cardiac catheterization via the radial artery. Group A was an accelerated protocol in which weaning was initiated 20 min after sheath removal. Group B was an adjusted protocol, in which weaning was dependent on the amount of anti-platelet or anti-coagulation used. All patients underwent radial artery ultrasound to demonstrate arterial patency. RESULTS: Baseline characteristics were similar in both groups, and PCI was performed in 36.7% of patients in Group A and 37.7% of patients in Group B. RAO occurred in 7.7% of patients overall, with no statistical difference between groups (Group A 5% versus Group B 10.1%, p-value = 0.337). Hematoma formation >5 cm in diameter occurred in 4.6% of patients in the overall cohort, without statistical difference between groups (Group A 5% versus Group B 4.3%, p-value = 1). The TR-Band duration was significantly shorter in Group A compared to Group B (112.9 ±â€¯50.7 versus 130.7 ±â€¯51.1 in minutes, respectively, p-value = 0.013). CONCLUSION: We have demonstrated an accelerated weaning protocol is simple to utilize for nursing staff without increased vascular site complications of RAO or hematoma formation.


Subject(s)
Cardiac Catheterization , Catheterization, Peripheral , Hemorrhage/prevention & control , Hemostatic Techniques/instrumentation , Radial Artery , Aged , Arterial Occlusive Diseases/diagnostic imaging , Arterial Occlusive Diseases/etiology , Arterial Occlusive Diseases/physiopathology , Catheterization, Peripheral/adverse effects , Female , Hematoma/etiology , Hemorrhage/etiology , Hemostatic Techniques/adverse effects , Humans , Male , Michigan , Middle Aged , Prospective Studies , Punctures , Radial Artery/diagnostic imaging , Radial Artery/physiopathology , Risk Factors , Time Factors , Treatment Outcome , Vascular Patency
10.
Cardiovasc Revasc Med ; 18(5): 328-331, 2017.
Article in English | MEDLINE | ID: mdl-28314674

ABSTRACT

OBJECTIVES: To evaluate the success rates and outcome of the hybrid algorithm for chronic total occlusion (CTO) percutaneous coronary intervention (PCI) by a single operator in two different clinical settings. METHODS: We compared 279 consecutive CTO PCIs performed by a single, high-volume operator using the hybrid algorithm in two different clinical settings. Data were collected through the PROGRESS CTO Registry. We compared 145 interventions performed in a community program (cohort A) with 134 interventions performed in a referral center (cohort B). RESULTS: Patient in cohort B had more complex lesions with higher J-CTO (3.0 vs. 3.41; p<0.001) and Progress CTO (1.5 vs.1.8, P=0.003) scores, more moderate to severe tortuosity (38% vs. 64%; p<0.001), longer total occlusion length (25 vs. 40mm; p<0.001) and higher prevalence of prior failed CTO PCI attempts (15% vs. 35%; p=0.001). Both technical (95% vs. 91%; p=0.266) and procedural (94% vs. 88%; p=0.088) success rates were similar between the two cohorts despite significantly different lesion complexity. Overall major adverse cardiovascular events were higher in cohort B (1.4% vs. 7.8%; p=0.012) without any significant difference in mortality (0.7% vs. 2.3%, p=0.351). CONCLUSIONS: In spite of higher lesion complexity in the setting of a quaternary-care referral center, use of the hybrid algorithm for CTO PCI enabled similarly high technical and procedural success rates as compared with those previously achieved by the same operator in a community-based program at the expense of a higher rate of MACE.


Subject(s)
Algorithms , Coronary Angiography , Coronary Occlusion/surgery , Percutaneous Coronary Intervention , Aged , Aged, 80 and over , Chronic Disease/therapy , Cohort Studies , Coronary Angiography/methods , Female , Humans , Male , Middle Aged , Percutaneous Coronary Intervention/methods , Registries , Risk Factors , Treatment Outcome
11.
Sci Rep ; 7: 40948, 2017 01 19.
Article in English | MEDLINE | ID: mdl-28102317

ABSTRACT

We present a combination of small-angle neutron scattering, deuterium labelling and contrast variation, temperature activation and fluorescence spectroscopy as a novel approach to obtain time-resolved, structural data individually from macromolecular complexes and their substrates during active biochemical reactions. The approach allowed us to monitor the mechanical unfolding of a green fluorescent protein model substrate by the archaeal AAA+ PAN unfoldase on the sub-minute time scale. Concomitant with the unfolding of its substrate, the PAN complex underwent an energy-dependent transition from a relaxed to a contracted conformation, followed by a slower expansion to its initial state at the end of the reaction. The results support a model in which AAA ATPases unfold their substrates in a reversible power stroke mechanism involving several subunits and demonstrate the general utility of this time-resolved approach for studying the structural molecular kinetics of multiple protein remodelling complexes and their substrates on the sub-minute time scale.


Subject(s)
ATPases Associated with Diverse Cellular Activities/chemistry , ATPases Associated with Diverse Cellular Activities/metabolism , Archaea/enzymology , Scattering, Small Angle , Green Fluorescent Proteins/metabolism , Protein Conformation , Protein Folding
12.
Biophys J ; 110(10): 2185-94, 2016 05 24.
Article in English | MEDLINE | ID: mdl-27224484

ABSTRACT

Water molecules in the immediate vicinity of biomacromolecules, including proteins, constitute a hydration layer characterized by physicochemical properties different from those of bulk water and play a vital role in the activity and stability of these structures, as well as in intermolecular interactions. Previous studies using solution scattering, crystallography, and molecular dynamics simulations have provided valuable information about the properties of these hydration shells, including modifications in density and ionic concentration. Small-angle scattering of x-rays (SAXS) and neutrons (SANS) are particularly useful and complementary techniques to study biomacromolecular hydration shells due to their sensitivity to electronic and nuclear scattering-length density fluctuations, respectively. Although several sophisticated SAXS/SANS programs have been developed recently, the impact of physicochemical surface properties on the hydration layer remains controversial, and systematic experimental data from individual biomacromolecular systems are scarce. Here, we address the impact of physicochemical surface properties on the hydration shell by a systematic SAXS/SANS study using three mutants of a single protein, green fluorescent protein (GFP), with highly variable net charge (+36, -6, and -29). The combined analysis of our data shows that the hydration shell is locally denser in the vicinity of acidic surface residues, whereas basic and hydrophilic/hydrophobic residues only mildly modify its density. Moreover, the data demonstrate that the density modifications result from the combined effect of residue-specific recruitment of ions from the bulk in combination with water structural rearrangements in their vicinity. Finally, we find that the specific surface-charge distributions of the different GFP mutants modulate the conformational space of flexible parts of the protein.


Subject(s)
Green Fluorescent Proteins/chemistry , Water/chemistry , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Green Fluorescent Proteins/genetics , Hydrophobic and Hydrophilic Interactions , Models, Molecular , Mutation , Neutron Diffraction , Scattering, Small Angle , Surface Properties , X-Ray Diffraction
13.
Acta Crystallogr D Biol Crystallogr ; 71(Pt 1): 57-66, 2015 Jan 01.
Article in English | MEDLINE | ID: mdl-25615860

ABSTRACT

Small-angle scattering (SAS) has witnessed a breathtaking renaissance and expansion over the past 15 years regarding the determination of biomacromolecular structures in solution. While important issues such as sample quality, good experimental practice and guidelines for data analysis, interpretation, presentation, publication and deposition are increasingly being recognized, crucial topics such as the uniqueness, precision and accuracy of the structural models obtained by SAS are still only poorly understood and addressed. The present article provides an overview of recent developments in these fields with a focus on the influence of complementary NMR restraints and of a hydration shell on the uniqueness of biomacromolecular models. As a first topic, the impact of incorporating NMR orientational restraints in addition to SAS distance restraints is discussed using a quantitative visual representation that illustrates how the possible conformational space of a two-body system is reduced as a function of the available data. As a second topic, the impact of a hydration shell on modelling parameters of a two-body system is illustrated, in particular on its inter-body distance. Finally, practical recommendations are provided to take both effects into account and promising future perspectives of SAS approaches are discussed.


Subject(s)
Models, Chemical , Nuclear Magnetic Resonance, Biomolecular/methods , Scattering, Small Angle , Water/chemistry
14.
RNA Biol ; 10(4): 579-89, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23603827

ABSTRACT

The RNA-binding protein TIAR is an mRNA-binding protein that acts as a translational repressor, particularly important under conditions of cellular stress. It binds to target mRNA and DNA via its RNA recognition motif (RRM) domains and is involved in both splicing regulation and translational repression via the formation of "stress granules." TIAR has also been shown to bind ssDNA and play a role in the regulation of transcription. Here we show, using surface plasmon resonance and nuclear magnetic resonance spectroscopy, specific roles of individual TIAR domains for high-affinity binding to RNA and DNA targets. We confirm that RRM2 of TIAR is the major RNA- and DNA-binding domain. However, the strong nanomolar affinity binding to U-rich RNA and T-rich DNA depends on the presence of the six amino acid residues found in the linker region C-terminal to RRM2. On its own, RRM1 shows preferred binding to DNA over RNA. We further characterize the interaction between RRM2 with the C-terminal extension and an AU-rich target RNA sequence using NMR spectroscopy to identify the amino acid residues involved in binding. We demonstrate that TIAR RRM2, together with its C-terminal extension, is the major contributor for the high-affinity (nM) interactions of TIAR with target RNA sequences.


Subject(s)
Amino Acid Motifs , DNA-Binding Proteins/metabolism , DNA/metabolism , RNA, Messenger/metabolism , RNA-Binding Proteins/chemistry , RNA-Binding Proteins/metabolism , Amino Acid Sequence , Binding Sites , DNA/genetics , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/genetics , Gene Expression Regulation , Humans , Magnetic Resonance Spectroscopy , Models, Molecular , Protein Binding , RNA Splicing , RNA, Messenger/genetics , RNA-Binding Proteins/genetics , Sequence Alignment , Surface Plasmon Resonance
15.
Am J Cardiol ; 110(1): 124-8, 2012 Jul 01.
Article in English | MEDLINE | ID: mdl-22482864

ABSTRACT

We evaluated preventive cardiology education in United States cardiology fellowship programs and their adherence to Core Cardiovascular Training Symposium training guidelines, which recommend 1 month of training, faculty with expertise, and clinical experience in cardiac rehabilitation, lipid disorder management, and diabetes management as a part of the prevention curricula. We sent an anonymous survey to United States cardiology program directors and their chief fellow. The survey assessed the program curricula, rotation structure, faculty expertise, obstacles, and recommended improvements. The results revealed that 24% of surveyed programs met the Core Cardiovascular Training Symposium guidelines with a dedicated 1-month rotation in preventive cardiology, 24% had no formalized training in preventive cardiology, and 30% had no faculty with expertise in preventive cardiology, which correlated with fewer rotations in prevention than those with specialized faculty (p = 0.009). Fellows rotated though the following experiences (% of programs): cardiac rehabilitation, 71%; lipid management, 37%; hypertension, 15%; diabetes, 7%; weight management/obesity, 6%; cardiac nutrition, 6%; and smoking cessation, 5%. The program directors cited "lack of time" as the greatest obstacle to providing preventive cardiology training and the chief fellows reported "lack of a developed curriculum" (p = 0.01). The most recommended improvement was for the American College of Cardiology to develop a web-based curriculum/module. In conclusion, most surveyed United States cardiology training programs currently do not adhere to basic preventive cardiovascular medicine Core Cardiovascular Training Symposium recommendations. Additional attention to developing curricular content and structure, including the creation of an American College of Cardiology on-line knowledge module might improve fellowship training in preventive cardiology.


Subject(s)
Cardiology/education , Curriculum/standards , Guidelines as Topic , Internship and Residency/standards , Cardiology/methods , Data Collection , Humans , Retrospective Studies , United States
16.
Nucleic Acids Res ; 39(3): 1117-30, 2011 Feb.
Article in English | MEDLINE | ID: mdl-21233170

ABSTRACT

TIAR and HuR are mRNA-binding proteins that play important roles in the regulation of translation. They both possess three RNA recognition motifs (RRMs) and bind to AU-rich elements (AREs), with seemingly overlapping specificity. Here we show using SPR that TIAR and HuR bind to both U-rich and AU-rich RNA in the nanomolar range, with higher overall affinity for U-rich RNA. However, the higher affinity for U-rich sequences is mainly due to faster association with U-rich RNA, which we propose is a reflection of the higher probability of association. Differences between TIAR and HuR are observed in their modes of binding to RNA. TIAR is able to bind deoxy-oligonucleotides with nanomolar affinity, whereas HuR affinity is reduced to a micromolar level. Studies with U-rich DNA reveal that TIAR binding depends less on the 2'-hydroxyl group of RNA than HuR binding. Finally we show that SAXS data, recorded for the first two domains of TIAR in complex with RNA, are more consistent with a flexible, elongated shape and not the compact shape that the first two domains of Hu proteins adopt upon binding to RNA. We thus propose that these triple-RRM proteins, which compete for the same binding sites in cells, interact with their targets in fundamentally different ways.


Subject(s)
Antigens, Surface/chemistry , DNA/chemistry , RNA-Binding Proteins/chemistry , RNA/chemistry , Adenine/analysis , Antigens, Surface/metabolism , DNA/metabolism , ELAV Proteins , ELAV-Like Protein 1 , Kinetics , Models, Molecular , Protein Binding , Protein Conformation , RNA/metabolism , RNA-Binding Proteins/metabolism , Scattering, Small Angle , Uracil/analysis , X-Ray Diffraction
17.
Atherosclerosis ; 205(1): 107-12, 2009 Jul.
Article in English | MEDLINE | ID: mdl-19144335

ABSTRACT

Matrix metalloproteinase-9 (MMP-9) has been proposed to be an important modulator of atherosclerotic plaque vulnerability. We generated a transgenic (tg) model expressing human proMMP-9 in macrophages, using the scavenger receptor enhancer/promoter A. This model was crossed into the double Apoe/Timp-1 knockout background. After 16 weeks of a high-fat diet, there were no significant changes in plaque size in the proximal aortas between the four groups of the study population (Apoe(-/-), Apoe(-/-)/MMP-9tg, Apoe(-/-)/Timp-1(-/-), and Apoe(-/-)/MMP-9tg/Timp-1(-/-)), and, in the Timp-1 knockout background, MMP-9 transgenic mice and control littermates had similar micro-aneurysm formation. However, lesions in Apoe(-/-)/MMP-9tg/Timp-1(-/-) mice contained significantly more collagen compared to the three other groups (P<0.005). Culture supernatants from elicited Apoe(-/-)/MMP-9tg/Timp-1(-/-) macrophages contained higher levels of active TGF-beta than the three other groups (P<0.05), suggesting that augmented collagen deposition resulted from an increase in TGF-beta activation due to transgenic MMP-9 in the Timp-1(-/-) background. This study indicates that, in human atherosclerosis, increased MMP-9 activity could up-regulate collagen deposition, possibly through TGF-beta activation.


Subject(s)
Atherosclerosis/enzymology , Atherosclerosis/genetics , Collagen/metabolism , Matrix Metalloproteinase 9/biosynthesis , Matrix Metalloproteinase 9/genetics , Transgenes , Animals , Aorta/pathology , Gene Expression Regulation , Humans , Macrophages/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Models, Biological , Transforming Growth Factor beta/metabolism
18.
J Thromb Thrombolysis ; 27(1): 18-23, 2009 Jan.
Article in English | MEDLINE | ID: mdl-17898930

ABSTRACT

BACKGROUND: The missed diagnosis of acute myocardial infarction has been studied in the Emergency Department, but few studies have investigated how often coronary ischemia is correctly identified in the outpatient setting. METHODS: This was a single center retrospective observational study of patients with Health Alliance Plan medical insurance hospitalized at a US tertiary center with acute myocardial infarction in 2004. Outpatient encounters in the 30 days preceding acute myocardial infarction were reviewed by two independent cardiologists for presenting symptoms and diagnostic decision-making in order to classify patient presentations as acute coronary ischemia, stable angina or neither. RESULTS: There were 331 patients with acute myocardial infarction, including 190 (57%) with a primary diagnosis of AMI and evaluated by a physician in the preceding 30 days. This group included 68 patients with 95 documented outpatient encounters by a primary care physician, cardiologist, or other internal medicine specialist which formed the final study population. Mean interval between these encounters and AMI was 17 +/- 11 days. Of these patients, 7 (10%) had symptoms of acute coronary ischemia, 5 (7%) had stable angina symptoms, and 56 (83%) had no symptoms of coronary ischemia at their outpatient encounters. Of the 7 patients with acute coronary ischemic symptoms, 5 were correctly identified and 2 were misidentified. CONCLUSION: A majority of patients with subsequent AMI visit an outpatient provider in the month preceding AMI. However, few present with symptoms of coronary ischemia in the outpatient setting (10%) and these symptoms are not always identified as such.


Subject(s)
Acute Coronary Syndrome/diagnosis , Ambulatory Care , Diagnostic Errors , Acute Coronary Syndrome/complications , Acute Coronary Syndrome/epidemiology , Aged , Aged, 80 and over , Algorithms , Ambulatory Care/statistics & numerical data , Cardiology , Chest Pain/etiology , Diagnostic Tests, Routine/statistics & numerical data , Dyspnea/etiology , Early Diagnosis , Female , Hospitals, Urban/statistics & numerical data , Humans , Internal Medicine , Male , Middle Aged , Nausea/etiology , Primary Health Care , Retrospective Studies , Syncope/etiology
19.
J Orthop Trauma ; 22(4): 258-63, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18404035

ABSTRACT

OBJECTIVES: With the development and popularization of minimally invasive surgical methods and implants for fracture fixation, it is increasingly important that the available implants are precontoured to the specific anatomic location for which they are designed. The objective of this study was to develop a noninvasive method and criteria for quantifying the fit of a distal periarticular medial tibia plate and to test the method on a small set of tibia models. METHODS: The undersurface of the plate was extracted from a digital model of the plate. The surface of the plate was fitted to 21 computer tomography (CT)-based 3-dimensional (3-D) models of human tibiae. Four criteria were defined that constitute an anatomic plate fit and subsequently were applied for the quantitative fit assessment. The fitting of the plate undersurface to the bone was entirely conducted in a virtual environment. RESULTS: An anatomic fit of the plate was achieved for 4 of the models (19%). The individual categories generated fits of 62% (n = 13) for the proximal end; 43% (n = 9) for the proximal angle; 57% (n = 12) for the middle distance; and 57% (n = 12) for a distal fit. CONCLUSIONS: Although for the 4 individual criteria plate fits of 43%-62% were achieved, a global/anatomic fit only occurred for 19% of the bone models. This outcome is likely a result of bone morphology variations, which exist in a random population sample combined with the effects of a nonoptimized plate shape. Recommendations for optimizing the fit of the plate are discussed.


Subject(s)
Computer-Aided Design , Fracture Fixation, Internal/instrumentation , Internal Fixators , Prosthesis Design/methods , Tibia/injuries , Absorbable Implants , Fractures, Bone/surgery , Humans , Prosthesis Fitting/methods
20.
Clin Cardiol ; 31(2): 67-71, 2008 Feb.
Article in English | MEDLINE | ID: mdl-18257021

ABSTRACT

BACKGROUND: Elevated troponin I has been associated with increased mortality in critically ill patients without acute coronary syndrome (ACS). However, the prognostic significance of troponin elevation in patients with diabetic ketoacidosis (DKA) without evident ACS has not been studied. METHODS: Retrospective study of all patients admitted to a U.S. tertiary center between 01/98 and 12/00 with DKA and had troponin I level measured. Patients with evidence of ACS or who met the American College of Cardiology/European Society of Cardiology (ACC/ESC) definition for myocardial infarction were excluded. Baseline characteristics, cardiac evaluation and 2 year major adverse coronary event (MACE) rate were compared between patients with positive and negative troponin. RESULTS: Ninety-six patients fulfilled the inclusion criteria of this study, 26 had positive troponin. There were no differences in baseline characteristics between the two groups. After a 2 year follow-up, there was significantly increased mortality in patients with elevated troponin (50.0% versus 27.1%, hazard-ratio (HR) 2.3, 95% confidence interval (CI) 1.2-4.8, p = 0.02). Patients with elevated troponin also had significantly increased MACE rate at 2 years (50.0% versus 28.6%, HR 2.6, 95% CI 1.3-5.3, p = 0.007) driven primarily by mortality. Using Cox Proportional Hazard Analysis, elevated troponin was a predictor of increased MACE after adjusting for confounding variables. (Adjusted HR 2.3, 95% CI 1.1-4.6, p = 0.02) CONCLUSIONS: Elevated troponin I in diabetic patients admitted with DKA identifies a group at very high risk for future cardiac events and mortality. Whether cardiac risk stratification of these patients will improve long term outcome remains to be studied.


Subject(s)
Coronary Artery Disease/blood , Coronary Artery Disease/diagnosis , Diabetic Ketoacidosis/blood , Troponin I/blood , Acute Coronary Syndrome , Biomarkers/blood , Cardiovascular Diseases/epidemiology , Cardiovascular Diseases/etiology , Coronary Artery Disease/etiology , Coronary Artery Disease/mortality , Diabetic Ketoacidosis/physiopathology , Female , Humans , Incidence , Kaplan-Meier Estimate , Male , Middle Aged , Predictive Value of Tests , Prognosis , Research Design , Retrospective Studies , Risk Factors , Severity of Illness Index , Time Factors
SELECTION OF CITATIONS
SEARCH DETAIL
...