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1.
AJR Am J Roentgenol ; 2024 Sep 04.
Artículo en Inglés | MEDLINE | ID: mdl-39230402

RESUMEN

Background: Retrospective studies evaluating artificial intelligence (AI) algorithms for intracranial hemorrhage (ICH) detection on noncontrast CT (NCCT) have shown promising results but lack prospective validation. Objective: To evaluate the impact on radiologists' real-world aggregate performance for ICH detection and report turnaround times for ICH-positive examinations of a radiology department's implementation of an AI triage and notification system for ICH detection on head NCCT examinations. Methods: This prospective single-center study included adult patients who underwent head NCCT examinations from May 12, 2021 to June 30, 2021 (phase 1) or September 30, 2021 to December 4, 2021 (phase 2). Before phase 1, the radiology department implemented a commercial AI triage system for ICH detection that processed head NCCT examinations and notified radiologists of positive results through a widget with a floating pop-up display. Examinations were interpreted by neuroradiologists or emergency radiologists, who evaluated examinations without and with AI assistance in phase 1 and phase 2, respectively. A panel of radiologists conducted a review process for all examinations with discordance between the radiology report and AI and a subset of remaining examinations, to establish the reference standard. Diagnostic performance and report turnaround times were compared using Pearson chi-square test and Wilcoxon rank-sum test, respectively. Bonferroni correction was used to account for five diagnostic performance metrics (adjusted significance threshold, .01 [α=.05/5]). Results: A total of 9954 examinations from 7371 patients (mean age, 54.8±19.8 years; 3773 female, 3598 male) were included. In phases 1 and 2, 19.8% (735/3716) and 21.9% (1368/6238) of examinations, respectively, were positive for ICH (P=.01). Radiologists without versus with AI showed no significant difference in accuracy (99.5% vs 99.2%), sensitivity (98.6% vs 98.9%), PPV (99.0% vs 99.7%), or NPV (99.7% vs 99.7%) (all P>.01); specificity was higher for radiologists without than with AI (99.8% vs 99.3%, respectively, P=.004). Mean report turnaround time for ICH-positive examinations was 147.1 minutes without AI versus 149.9 minutes with AI (P=.11). Conclusion: An AI triage system for ICH detection did not improve radiologists' diagnostic performance or report turnaround times. Clinical Impact: This large prospective real-world study does not support use of AI assistance for ICH detection.

2.
Childs Nerv Syst ; 40(1): 197-204, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-37864710

RESUMEN

BACKGROUND: Dural arteriovenous fistulas (dAVF) are arteriovenous shunts in communication with the dural vasculature in the brain or spine. Apart from single-center series, risk factors and treatment outcomes for pediatric dAVFs are largely undescribed. METHODS: We performed a systematic literature review of pediatric (< 18 years at diagnosis) intracranial and spinal dAVF according to PRISMA guidelines. We queried PubMed, CINAHL, SCOPUS, and Embase databases without time/date restriction. Search strings included a variety of MeSH keywords relating to dural AV fistulas in combination with MeSH keywords related to pediatric cases (see Appendix). Manuscripts describing patients diagnosed with dural sinus malformations or pial AVF were excluded. RESULTS: We identified 61 studies describing 69 individual patients. Overall, dAVF were more common in males (55.1%) with a mean age of diagnosis (5.17 ± 4.42 years). Approximately 20.2% of patients presented with cardiovascular disease (CVD), and 31.9% were discovered incidentally on neuroimaging studies. Transverse-sigmoid junction was the most common location (17.3%). Ninety-three percent (64 patients) were treated, most commonly using endovascular embolization (68.1%) followed by surgery (8.7%) and radiosurgery (2.9%). Almost half (43.8%) of dAVFs were completely obliterated. Of the 64 procedures, there were 19 neurological complications (29.7%) of varying severity where 12.5% were considered transient (i.e., pseudomeningocele) and 17.2% permanent (i.e., mortality secondary to acute sinus thrombosis, etc.). CONCLUSION: There is a paucity of information on pediatric dAVFs. This systematic review summarizes the published cases of dAVFs in the pediatric population. While the rate of missing data is high, there is publication bias, and precise details regarding complications are difficult to ascertain, this review serves as a descriptive summary of pediatric dAVFs.


Asunto(s)
Fístula Arteriovenosa , Malformaciones Vasculares del Sistema Nervioso Central , Embolización Terapéutica , Radiocirugia , Masculino , Humanos , Niño , Lactante , Preescolar , Resultado del Tratamiento , Embolización Terapéutica/métodos , Malformaciones Vasculares del Sistema Nervioso Central/diagnóstico por imagen , Malformaciones Vasculares del Sistema Nervioso Central/terapia , Fístula Arteriovenosa/diagnóstico por imagen , Fístula Arteriovenosa/terapia , Fístula Arteriovenosa/etiología
3.
Childs Nerv Syst ; 40(6): 1721-1729, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38506930

RESUMEN

INTRODUCTION: Pediatric non-galenic pial arteriovenous fistulas (pAVFs) are rare vascular malformations that are characterized by a pial arterial-venous connection without an intervening capillary bed. Outcomes and treatment strategies for pAVFs are highly individualized, owing to the rarity of the disease and lack of large-scale data guiding optimal treatment approaches. METHODS: We performed a systematic review of pediatric patients (< 18 years at diagnosis) diagnosed with a pAVF by digital subtraction angiogram (DSA). The demographics, treatment modalities, and outcomes were documented for each patient and clinical outcome data was collected. Descriptive information stratified by outcome scores were classified as follows: 1 = excellent (no deficit and full premorbid activity), 2 = good (mild deficit and full premorbid activity), 3 = fair (moderate deficit and impaired activity), 4 = poor (severe deficit and dependent on others), 5 = death. RESULTS: A total of 87 studies involving 231 patients were identified. Median age at diagnosis was 3 years (neonates to 18 years). There was slight male preponderance (55.4%), and 150 subjects (81.1%*) experienced excellent outcomes after treatment. Of the 189 patients treated using endovascular approaches, 80.3% experienced excellent outcomes and of the 15 patients surgically treated subjects 75% had an excellent outcome. The highest rate of excellent outcomes was achieved in patients treated with Onyx (95.2%) and other forms of EvOH (100%). High output heart failure and comorbid vascular lesions tended to result in worse outcomes, with only 54.2% and 68% of subjects experiencing an excellent outcome, respectively. *Outcomes were reported in only 185 patients. CONCLUSION: pAVFs are rare lesions, necessitating aggregation of patient data to inform natural history and optimal treatment strategies. This review summarizes the current literature on pAVF in children, where children presenting with heart failure as a result of high flow through the lesion were less likely to experience an excellent outcome. Prospective, large-scale studies would further characterize pediatric pAVFs and enable quantitative analysis of outcomes to inform best treatment practices.


Asunto(s)
Fístula Arteriovenosa , Piamadre , Humanos , Niño , Fístula Arteriovenosa/cirugía , Fístula Arteriovenosa/diagnóstico por imagen , Fístula Arteriovenosa/terapia , Piamadre/irrigación sanguínea , Preescolar , Adolescente , Lactante , Femenino , Recién Nacido , Resultado del Tratamiento , Masculino , Malformaciones Arteriovenosas Intracraneales/terapia , Malformaciones Arteriovenosas Intracraneales/diagnóstico por imagen , Malformaciones Arteriovenosas Intracraneales/cirugía
4.
Neurosurg Rev ; 47(1): 116, 2024 Mar 14.
Artículo en Inglés | MEDLINE | ID: mdl-38483647

RESUMEN

BACKGROUND: The Woven EndoBridge (WEB) devices have been used for treating wide neck bifurcation aneurysms (WNBAs) with several generational enhancements to improve clinical outcomes. The original device dual-layer (WEB DL) was replaced by a single-layer (WEB SL) device in 2013. This study aimed to compare the effectiveness and safety of these devices in managing intracranial aneurysms. METHODS: A multicenter cohort study was conducted, and data from 1,289 patients with intracranial aneurysms treated with either the WEB SL or WEB DL devices were retrospectively analyzed. Propensity score matching was utilized to balance the baseline characteristics between the two groups. Outcomes assessed included immediate occlusion rate, complete occlusion at last follow-up, retreatment rate, device compaction, and aneurysmal rupture. RESULTS: Before propensity score matching, patients treated with the WEB SL had a significantly higher rate of complete occlusion at the last follow-up and a lower rate of retreatment. After matching, there was no significant difference in immediate occlusion rate, retreatment rate, or device compaction between the WEB SL and DL groups. However, the SL group maintained a higher rate of complete occlusion at the final follow-up. Regression analysis showed that SL was associated with higher rates of complete occlusion (OR: 0.19; CI: 0.04 to 0.8, p = 0.029) and lower rates of retreatment (OR: 0.12; CI: 0 to 4.12, p = 0.23). CONCLUSION: The WEB SL and DL devices demonstrated similar performances in immediate occlusion rates and retreatment requirements for intracranial aneurysms. The SL device showed a higher rate of complete occlusion at the final follow-up.


Asunto(s)
Embolización Terapéutica , Procedimientos Endovasculares , Aneurisma Intracraneal , Humanos , Resultado del Tratamiento , Aneurisma Intracraneal/cirugía , Aneurisma Intracraneal/etiología , Embolización Terapéutica/efectos adversos , Puntaje de Propensión , Estudios Retrospectivos , Estudios de Cohortes , Procedimientos Endovasculares/efectos adversos
5.
Neurosurg Rev ; 47(1): 374, 2024 Jul 31.
Artículo en Inglés | MEDLINE | ID: mdl-39083214

RESUMEN

The Woven EndoBridge (WEB) device is primarily used for treating wide-neck intracranial bifurcation aneurysms under 10 mm. Limited data exists on its efficacy for large aneurysms. We aim to assess angiographic and clinical outcomes of the WEB device in treating large versus small aneurysms. We conducted a retrospective review of the WorldWide WEB Consortium database, from 2011 to 2022, across 30 academic institutions globally. Propensity score matching (PSM) was employed to compare small and large aneurysms on baseline characteristics. A total of 898 patients were included. There was no significant difference observed in clinical presentations, smoking status, pretreatment mRS, presence of multiple aneurysms, bifurcation location, or prior treatment between the two groups. After PSM, 302 matched pairs showed significantly lower last follow-up adequate occlusion rates (81% vs 90%, p = 0.006) and higher retreatment rates (12% vs 3.6%, p < 0.001) in the large aneurysm group. These findings may inform treatment decisions and patient counseling. Future studies are needed to further explore this area.


Asunto(s)
Procedimientos Endovasculares , Aneurisma Intracraneal , Puntaje de Propensión , Humanos , Aneurisma Intracraneal/terapia , Aneurisma Intracraneal/cirugía , Masculino , Femenino , Persona de Mediana Edad , Estudios Retrospectivos , Anciano , Procedimientos Endovasculares/métodos , Resultado del Tratamiento , Adulto , Embolización Terapéutica/métodos
6.
J Stroke Cerebrovasc Dis ; 33(11): 107897, 2024 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-39069148

RESUMEN

INTRODUCTION: The Woven EndoBridge (WEB) device is emerging as a novel therapy for intracranial aneurysms, but its use for off-label indications requires further study. Using machine learning, we aimed to develop predictive models for complete occlusion after off-label WEB treatment and to identify factors associated with occlusion outcomes. METHODS: This multicenter, retrospective study included 162 patients who underwent off-label WEB treatment for intracranial aneurysms. Baseline, morphological, and procedural variables were utilized to develop machine-learning models predicting complete occlusion. Model interpretation was performed to determine significant predictors. Ordinal regression was also performed with occlusion status as an ordinal outcome from better (Raymond Roy Occlusion Classification [RROC] grade 1) to worse (RROC grade 3) status. Odds ratios (OR) with 95 % confidence intervals (CI) were reported. RESULTS: The best performing model achieved an AUROC of 0.8 for predicting complete occlusion. Larger neck diameter and daughter sac were significant independent predictors of incomplete occlusion. On multivariable ordinal regression, higher RROC grades (OR 1.86, 95 % CI 1.25-2.82), larger neck diameter (OR 1.69, 95 % CI 1.09-2.65), and presence of daughter sacs (OR 2.26, 95 % CI 0.99-5.15) were associated with worse aneurysm occlusion after WEB treatment, independent of other factors. CONCLUSION: This study found that larger neck diameter and daughter sacs were associated with worse occlusion after WEB therapy for aneurysms. The machine learning approach identified anatomical factors related to occlusion outcomes that may help guide patient selection and monitoring with this technology. Further validation is needed.


Asunto(s)
Procedimientos Endovasculares , Aneurisma Intracraneal , Aprendizaje Automático , Uso Fuera de lo Indicado , Humanos , Aneurisma Intracraneal/terapia , Aneurisma Intracraneal/diagnóstico por imagen , Masculino , Femenino , Estudios Retrospectivos , Persona de Mediana Edad , Resultado del Tratamiento , Anciano , Factores de Riesgo , Procedimientos Endovasculares/instrumentación , Procedimientos Endovasculares/efectos adversos , Prótesis Vascular , Diseño de Prótesis , Técnicas de Apoyo para la Decisión , Implantación de Prótesis Vascular/instrumentación , Implantación de Prótesis Vascular/efectos adversos , Adulto , Toma de Decisiones Clínicas , Medición de Riesgo
7.
Biomacromolecules ; 24(3): 1388-1399, 2023 03 13.
Artículo en Inglés | MEDLINE | ID: mdl-36796007

RESUMEN

Encapsulins are microbial protein nanocages capable of efficient self-assembly and cargo enzyme encapsulation. Due to their favorable properties, including high thermostability, protease resistance, and robust heterologous expression, encapsulins have become popular bioengineering tools for applications in medicine, catalysis, and nanotechnology. Resistance against physicochemical extremes like high temperature and low pH is a highly desirable feature for many biotechnological applications. However, no systematic search for acid-stable encapsulins has been carried out, while the influence of pH on encapsulin shells has so far not been thoroughly explored. Here, we report on a newly identified encapsulin nanocage from the acid-tolerant bacterium Acidipropionibacterium acidipropionici. Using transmission electron microscopy, dynamic light scattering, and proteolytic assays, we demonstrate its extreme acid tolerance and resilience against proteases. We structurally characterize the novel nanocage using cryo-electron microscopy, revealing a dynamic five-fold pore that displays distinct "closed" and "open" states at neutral pH but only a singular "closed" state under strongly acidic conditions. Further, the "open" state exhibits the largest pore in an encapsulin shell reported to date. Non-native protein encapsulation capabilities are demonstrated, and the influence of external pH on internalized cargo is explored. Our results expand the biotechnological application range of encapsulin nanocages toward potential uses under strongly acidic conditions and highlight pH-responsive encapsulin pore dynamics.


Asunto(s)
Bacterias , Proteínas Bacterianas , Proteínas Bacterianas/química , Microscopía por Crioelectrón , Bacterias/metabolismo , Biotecnología , Nanotecnología
8.
Radiology ; 304(2): 372-382, 2022 08.
Artículo en Inglés | MEDLINE | ID: mdl-35438564

RESUMEN

Background The Woven EndoBridge (WEB) device was explicitly designed for wide-neck intracranial bifurcation aneurysms. Small-scale reports have evaluated the off-label use of WEB devices for the treatment of sidewall aneurysms, with promising outcomes. Purpose To compare the angiographic and clinical outcomes of the WEB device for the treatment of sidewall aneurysms compared with the treatment of bifurcation aneurysms. Materials and Methods A retrospective review of the WorldWideWEB Consortium, a synthesis of retrospective databases spanning from January 2011 to June 2021 at 22 academic institutions in North America, South America, and Europe, was performed to identify patients with intracranial aneurysms treated with the WEB device. Characteristics and outcomes were compared between bifurcation and sidewall aneurysms. Propensity score matching (PSM) was used to match by age, pretreatment ordinal modified Rankin Scale score, ruptured aneurysms, location of aneurysm, multiple aneurysms, prior treatment, neck, height, dome width, daughter sac, and incorporated branch. Results A total of 683 intracranial aneurysms were treated using the WEB device in 671 patients (median age, 61 years [IQR, 53-68 years]; male-to-female ratio, 1:2.5). Of those, 572 were bifurcation aneurysms and 111 were sidewall aneurysms. PSM was performed, resulting in 91 bifurcation and sidewall aneurysms pairs. No significant difference was observed in occlusion status at last follow-up, deployment success, or complication rates between the two groups. Conclusion No significantly different outcomes were observed following the off-label use of the Woven EndoBridge, or WEB, device for treatment of sidewall aneurysms compared with bifurcation aneurysms. The correct characterization of the sidewall aneurysm location, neck angle, and size is crucial for successful treatment and lower retreatment rate. © RSNA, 2022 See also the editorial by Hetts in this issue.


Asunto(s)
Aneurisma Roto , Embolización Terapéutica , Procedimientos Endovasculares , Aneurisma Intracraneal , Aneurisma Roto/terapia , Embolización Terapéutica/métodos , Procedimientos Endovasculares/métodos , Femenino , Humanos , Aneurisma Intracraneal/diagnóstico por imagen , Aneurisma Intracraneal/cirugía , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Resultado del Tratamiento
9.
Biotechnol Bioeng ; 118(1): 491-505, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-32918485

RESUMEN

Compartmentalization is an essential feature of all cells. It allows cells to segregate and coordinate physiological functions in a controlled and ordered manner. Different mechanisms of compartmentalization exist, with the most relevant to prokaryotes being encapsulation via self-assembling protein-based compartments. One widespread example of such is that of encapsulins-cage-like protein nanocompartments able to compartmentalize specific reactions, pathways, and processes in bacteria and archaea. While still relatively nascent bioengineering tools, encapsulins exhibit many promising characteristics, including a number of defined compartment sizes ranging from 24 to 42 nm, straightforward expression, the ability to self-assemble via the Hong Kong 97-like fold, marked physical robustness, and internal and external handles primed for rational genetic and molecular manipulation. Moreover, encapsulins allow for facile and specific encapsulation of native or heterologous cargo proteins via naturally or rationally fused targeting peptide sequences. Taken together, the attributes of encapsulins promise substantial customizability and broad usability. This review discusses recent advances in employing engineered encapsulins across various fields, from their use as bionanoreactors to targeted delivery systems and beyond. A special focus will be provided on the rational engineering of encapsulin systems and their potential promise as biomolecular research tools.


Asunto(s)
Archaea , Proteínas Arqueales , Bacterias , Proteínas Bacterianas , Nanoestructuras/química , Péptidos , Ingeniería de Proteínas , Proteínas Recombinantes de Fusión , Archaea/química , Archaea/genética , Archaea/metabolismo , Proteínas Arqueales/química , Proteínas Arqueales/genética , Proteínas Arqueales/metabolismo , Bacterias/química , Bacterias/genética , Bacterias/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Péptidos/química , Péptidos/genética , Péptidos/metabolismo , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo
10.
Angew Chem Int Ed Engl ; 60(47): 25034-25041, 2021 11 15.
Artículo en Inglés | MEDLINE | ID: mdl-34532937

RESUMEN

Protein nanocages play crucial roles in sub-cellular compartmentalization and spatial control in all domains of life and have been used as biomolecular tools for applications in biocatalysis, drug delivery, and bionanotechnology. The ability to control their assembly state under physiological conditions would further expand their practical utility. To gain such control, we introduced a peptide capable of triggering conformational change at a key structural position in the largest known encapsulin nanocompartment. We report the structure of the resulting engineered nanocage and demonstrate its ability to disassemble and reassemble on demand under physiological conditions. We demonstrate its capacity for in vivo encapsulation of proteins of choice while also demonstrating in vitro cargo loading capabilities. Our results represent a functionally robust addition to the nanocage toolbox and a novel approach for controlling protein nanocage disassembly and reassembly under mild conditions.


Asunto(s)
Nanopartículas/química , Ingeniería de Proteínas , Proteínas/química , Modelos Moleculares
11.
AIDS Behav ; 24(3): 682-696, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-30868447

RESUMEN

Access to safe and effective sexual healthcare services for transgender and male sex workers (TMSW) is a human right. Globally, TMSW experience a higher prevalence of human immunodeficiency virus (HIV) and sexually transmitted infections than the general population or other sex workers, suggesting the existence of unique challenges for this group when accessing healthcare. A systematic database search identified 22 qualitative papers addressing barriers to accessing sexual healthcare services for TMSW. These papers were critically evaluated for adherence to best practice standards for qualitative research and research with sex workers. A coding process identified five themes. Stigma was the predominant barrier, and was divided into stigma related to sexuality, gender identity, HIV status, sex worker status, and internalised stigma. Other barriers were confidentiality concerns, sexual health literacy, fatalism, and structural barriers. Each of these themes were informed by the wider context of stigma. The literature presents a complex syndemic of social disadvantage and exclusion acting to produce and reinforce health disparities related to sexual health and access to screening and treatment for TMSW.


Asunto(s)
Accesibilidad a los Servicios de Salud , Disparidades en Atención de Salud , Trabajadores Sexuales , Salud Sexual , Estigma Social , Personas Transgénero , Confidencialidad , Femenino , Infecciones por VIH/diagnóstico , Infecciones por VIH/prevención & control , Infecciones por VIH/terapia , Alfabetización en Salud , Humanos , Masculino , Tamizaje Masivo , Distancia Psicológica , Investigación Cualitativa , Conducta Sexual , Sexualidad , Enfermedades de Transmisión Sexual/diagnóstico , Enfermedades de Transmisión Sexual/prevención & control , Enfermedades de Transmisión Sexual/terapia , Sindémico
12.
Biochem Biophys Res Commun ; 516(2): 333-338, 2019 08 20.
Artículo en Inglés | MEDLINE | ID: mdl-31204053

RESUMEN

Herein we report the first structure of topoisomerase I determined from the gram-positive bacterium, S. mutans. Bacterial topoisomerase I is an ATP-independent type 1A topoisomerase that uses the inherent torsional strain within hyper-negatively supercoiled DNA as an energy source for its critical function of DNA relaxation. Interest in the enzyme has gained momentum as it has proven to be essential in various bacterial organisms. In order to aid in further biochemical characterization, the apo 65-kDa amino-terminal fragment of DNA topoisomerase I from the gram-positive model organism Streptococcus mutans was crystalized and a three-dimensional structure was determined to 2.06 Šresolution via x-ray crystallography. The overall structure illustrates the four classic major domains that create the traditional topoisomerase I "lock" formation comprised of a sizable toroidal aperture atop what is considered to be a highly dynamic body. A catalytic tyrosine residue resides at the interface between two domains and is known to form a 5' phosphotyrosine DNA-enzyme intermediate during transient single-stranded cleavage required for enzymatic relaxation of hyper negative DNA supercoils. Surrounding the catalytic tyrosine residue is the remainder of the highly conserved active site. Within 5 Šfrom the catalytic center, only one dissimilar residue is observed between topoisomerase I from S. mutans and the gram-negative model organism E. coli. Immediately adjacent to the conserved active site, however, S. mutans topoisomerase I displays a somewhat unique nine residue loop extension not present in any bacterial topoisomerase I structures previously determined other than that of an extremophile.


Asunto(s)
ADN-Topoisomerasas de Tipo I/química , Streptococcus mutans/enzimología , Secuencia de Aminoácidos , Cristalización , Cristalografía por Rayos X , Modelos Moleculares
13.
Protein Expr Purif ; 124: 32-40, 2016 08.
Artículo en Inglés | MEDLINE | ID: mdl-27117979

RESUMEN

Type IA topoisomerases represent promising antibacterial drug targets. Data exists suggesting that the two bacterial type IA topoisomerase enzymes-topoisomerase I and topoisomerase III-share an overlapping biological role. Furthermore, topoisomerase I has been shown to be essential for the survival of certain organisms lacking topoisomerase III. With this in mind, it is plausible that topoisomerase I may represent a potential target for selective antibacterial drug development. As many reported bacterial topoisomerase I purification protocols have either suffered from relatively low yield, numerous steps, or a simple failure to report target protein yield altogether, a high-yield and high-purity bacterial topoisomerase I expression and purification protocol is highly desirable. The goal of this study was therefore to optimize the expression and purification of topoisomerase I from Streptococcus mutans, a clinically relevant organism that plays a significant role in oral and extra-oral infection, in order to quickly and easily attain the requisite quantities of pure target enzyme suitable for use in assay development, compound library screening, and carrying out further structural and biochemical characterization analyses. Herein we report the systematic implementation and analysis of various expression and purification techniques leading to the development and optimization of a rapid and straightforward protocol for the auto-induced expression and two-step, affinity tag purification of Streptococcus mutans topoisomerase I yielding >20 mg/L of enzyme at over 95% purity.


Asunto(s)
Proteínas Bacterianas , ADN-Topoisomerasas de Tipo I , Expresión Génica , Streptococcus mutans/enzimología , Proteínas Bacterianas/antagonistas & inhibidores , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , ADN-Topoisomerasas de Tipo I/biosíntesis , ADN-Topoisomerasas de Tipo I/química , ADN-Topoisomerasas de Tipo I/aislamiento & purificación
14.
Magn Reson Med ; 71(2): 635-44, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-23504992

RESUMEN

PURPOSE: Dynamic contrast-enhanced imaging provides unique physiological information, notably the endothelial permeability (K(trans)), and may improve the diagnosis and management of multiple pathologies. Current acquisition methods provide limited spatial-temporal resolution and field-of-view, often preventing characterization of the entire pathology and precluding measurement of the arterial input function. We present a method for highly accelerated dynamic imaging and demonstrate its utility for dynamic contrast-enhanced modeling. METHODS: We propose a novel Poisson ellipsoid sampling scheme and enforce multiple spatial and temporal l1-norm constraints during image reconstruction. Retrospective and prospective analyses were performed to validate the approach. RESULTS: Retrospectively, no mean bias or diverging trend was observed as the acceleration rate was increased from 3× to 18×; less than 10% error was measured in K(trans) at any individual rates in this range. Prospectively accelerated images at a rate of 36× enabled full brain coverage with 0.94 × 0.94 × 1.9 mm(3) spatial and 4.1 s temporal resolutions. Images showed no visible degradation and provided accurate K(trans) values when compared to a clinical population. CONCLUSION: Highly accelerated dynamic MRI using compressed sensing and parallel imaging provides accurate permeability modeling and enables full brain, high resolution acquisitions.


Asunto(s)
Imagen por Resonancia Magnética/métodos , Encéfalo/anatomía & histología , Neoplasias Encefálicas/diagnóstico , Medios de Contraste , Endotelio/fisiología , Humanos , Esclerosis Múltiple/diagnóstico , Permeabilidad , Estudios Prospectivos , Estudios Retrospectivos
15.
Interv Neuroradiol ; : 15910199241255154, 2024 May 27.
Artículo en Inglés | MEDLINE | ID: mdl-38803217

RESUMEN

Pediatric neurointervention differs from the adult in several important respects. Here we describe a modern approach to readily acquire diagnostic quality images of children. Preparation, access, angiogragraphy and closure have evolved along with new knowledge and technology. This timely "how I do it" series addresses each topic utilizing literature review and our own experience over 35 years.

16.
Interv Neuroradiol ; : 15910199241275710, 2024 Aug 25.
Artículo en Inglés | MEDLINE | ID: mdl-39183554

RESUMEN

Pediatric arteriovenous shunts can be challenging to embolize and the consequences of excess venous penetration disastrous. Moreover, the congenital nature of most shunts necessitates neonatal intervention whenever (medically refractory) heart failure occurs. Here, we describe current endovascular treatment strategies based on personal experience and literature review. While disparate, arteriovenous malformation, dural arteriovenous fistula (AVF), pial AVF, and vein of Galen aneurysmal malformation share unifying features such as high output heart failure and venous hypertension. This tutorial is divided into passages on Goals, Access, and Treatment which respect differences among pediatric arteriovenous shunts while maintaining a narrative economy.

17.
Nat Commun ; 15(1): 2558, 2024 Mar 22.
Artículo en Inglés | MEDLINE | ID: mdl-38519509

RESUMEN

Encapsulins are self-assembling protein nanocompartments capable of selectively encapsulating dedicated cargo proteins, including enzymes involved in iron storage, sulfur metabolism, and stress resistance. They represent a unique compartmentalization strategy used by many pathogens to facilitate specialized metabolic capabilities. Encapsulation is mediated by specific cargo protein motifs known as targeting peptides (TPs), though the structural basis for encapsulation of the largest encapsulin cargo class, dye-decolorizing peroxidases (DyPs), is currently unknown. Here, we characterize a DyP-containing encapsulin from the enterobacterial pathogen Klebsiella pneumoniae. By combining cryo-electron microscopy with TP and TP-binding site mutagenesis, we elucidate the molecular basis for cargo encapsulation. TP binding is mediated by cooperative hydrophobic and ionic interactions as well as shape complementarity. Our results expand the molecular understanding of enzyme encapsulation inside protein nanocompartments and lay the foundation for rationally modulating encapsulin cargo loading for biomedical and biotechnological applications.


Asunto(s)
Proteínas Bacterianas , Peroxidasa , Proteínas Bacterianas/metabolismo , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/metabolismo , Microscopía por Crioelectrón , Peroxidasas/metabolismo
18.
MicroPubl Biol ; 20242024.
Artículo en Inglés | MEDLINE | ID: mdl-38545437

RESUMEN

The anticancer drug Gefitinib is a tyrosine kinase inhibitor with selectivity for the Epidermal Growth Factor Receptor (EGFR/ErbB1). As the C. elegans EGF receptor LET-23 shares notable structural homology over its kinase domain with human EGFR, we wished to examine whether Gefitinib treatment can interfere with LET-23-dependent processes. We show that Gefitinib disrupts C. elegans stress-induced sleep (SIS) but does not impact EGF overexpression-induced sleep nor vulva induction. These findings indicate that Gefitinib does not interfere with LET-23 signaling and impairs SIS through an off-target mechanism.

19.
J Clin Imaging Sci ; 14: 19, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38975060

RESUMEN

Magnetic resonance imaging (MRI) of the brain has been implemented to evaluate multiple intracranial pathologies. Non-contrast T2-weighted images are a routinely acquired sequence in almost all neuroimaging protocols. It is not uncommon to encounter various cerebrovascular lesions incidentally on brain imaging. Neuroradiologists should evaluate the routine T2-weighted images for incidental cerebrovascular lesions, irrespective of the primary indication of the study. Vascular structures typically demonstrate a low signal flow-void on the T2-weighted images. In our experience, large cerebrovascular abnormalities are easily visible to a typical neuroradiologist. In this article, we present the spectrum of the characteristic imaging appearance of various intracranial cerebrovascular lesions on routine non-contrast T2-weighted MRI. These include aneurysm, arteriovenous malformation, arterial occlusion, capillary telangiectasia, cavernous malformation, dural arteriovenous fistula, moyamoya, proliferative angiopathy, and vein of Galen malformation.

20.
Artículo en Inglés | MEDLINE | ID: mdl-39492799

RESUMEN

Open dumping and burning of solid waste are widely practiced in underserved communities lacking access to solid waste management facilities; however, the generation of microplastics from these sites has been overlooked. We report elevated concentrations of microplastics (MPs) in soil of three solid waste open dump and burn sites: a single-family site in Tuttle, Oklahoma, USA, and two community-wide sites in Crow Agency and Lodge Grass, Montana, USA. We extracted, quantified, and characterized MPs from two soil depths (0-9 cm and 9-18 cm). The average of abundance of particles found at community-wide sites three sites (18, 460 particles kg-1 soil) equals or exceeds reported concentrations from currently understood sources of MPs including biosolids application and other agricultural practices. Attenuated total reflectance Fourier transformed infrared (ATR-FTIR) identified polyethylene as the dominant polymer across all sites (46.2-84.8%). We also detected rayon (≤11.5%), polystyrene (up to 11.5%), polyethylene terephthalate (≤5.1), polyvinyl chloride (≤4.4%), polyester (≤3.1), and acrylic (≤2.2%). Burned MPs accounted for 76.3 to 96.9% of the MPs found in both community wide dumping sites. These results indicate that solid waste dumping and burning activities are a major source of thermally oxidized MPs for the surrounding terrestrial environment with potential to negatively affect underserved communities.

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