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1.
BMC Musculoskelet Disord ; 25(1): 157, 2024 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-38373917

RESUMEN

BACKGROUND: Anterior cruciate ligament (ACL) rupture is a common orthopedic injury, occurring in roughly 68.6 per 100,000 persons annually, with the primary treatment option being ACL reconstruction. However, debate remains about the appropriate graft type for restoring the native biomechanical properties of the knee. Furthermore, plastic graft elongation may promote increased knee laxity and instability without rupture. This study aims to investigate the plastic properties of common ACL-R graft options. METHODS: Patellar tendon (PT), hamstring tendon (HT), and quadriceps tendon (QT) grafts were harvested from 11 cadaveric knees (6 male and 5 female) with a mean age of 71(range 55-81). All grafts were mechanically tested under uniaxial tension until failure to determine each graft's elastic and plastic biomechanical properties. RESULTS: Mechanically, the QT graft was the weakest, exhibiting the lowest failure force and the lowest failure stress (QT < HT, p = 0.032). The PT was the stiffest of the grafts, having a significantly higher stiffness (PT > QT, p = 0.0002) and Young's modulus (PT > QT, p = 0.001; PT > HT, p = 0.041). The HT graft had the highest plastic elongation at 4.01 ± 1.32 mm (HT > PT, p = 0.002). The post-yield behavior of the HT tendon shows increased energy storage capabilities with the highest plastic energy storage (HT > QT, p = 0.012) and the highest toughness (HT > QT, p = 0.032). CONCLUSION: Our study agrees with prior studies indicating that the failure load of all grafts is above the requirements for everyday activities. However, grafts may be susceptible to yielding before failure during daily activities. This may result in the eventual loss of functionality for the neo-ACL, resulting in increased knee laxity and instability.


Asunto(s)
Lesiones del Ligamento Cruzado Anterior , Ligamento Rotuliano , Masculino , Humanos , Femenino , Anciano , Ligamento Cruzado Anterior/cirugía , Autoinjertos/cirugía , Trasplante Autólogo , Articulación de la Rodilla/cirugía , Ligamento Rotuliano/cirugía , Lesiones del Ligamento Cruzado Anterior/cirugía
2.
BMC Musculoskelet Disord ; 24(1): 263, 2023 Apr 04.
Artículo en Inglés | MEDLINE | ID: mdl-37016368

RESUMEN

BACKGROUND: The incidence of periprosthetic femur fracture (PPFF) in the setting of total hip arthroplasty (THA) is steadily increasing. We seek to address whether there is a difference in outcomes between Vancouver B fracture types managed with ORIF when the original stem was a press-fit stem versus a cemented stem. METHODS: In this retrospective cohort study at a level 1 trauma center, we identified 136 patients over 65 years-of-age with Vancouver B-type fractures sustained between 2005 and 2019. Patients were treated by ORIF and had either cemented or press-fit stems prior to their injury. Outcomes were subsidence of the femoral implant, time to full weight bearing, rate of the hip implant revision, estimated blood loss (EBL), postoperative complications, and the one-year mortality rate. RESULTS: A total of 103 (75.7%) press-fit and 33 (24.3%) cemented patients were reviewed. Patient baseline characteristics, Vancouver fracture sub-types, and implant characteristics were not found to be significantly different between groups. The difference in subsidence rates, postoperative complications, and time to weight bearing were not significantly different between groups. EBL and one-year mortality rate were significantly higher in the cemented group. CONCLUSIONS: In geriatric patients with Vancouver B type periprosthetic fractures managed with ORIF, patients with an originally press fit stem may have lower mortality, lower estimated blood loss, and similar subsidence and hospital length of stays when compared to those with a cemented stem.


Asunto(s)
Artroplastia de Reemplazo de Cadera , Fracturas del Fémur , Prótesis de Cadera , Fracturas Periprotésicas , Humanos , Anciano , Fracturas Periprotésicas/epidemiología , Fracturas Periprotésicas/etiología , Fracturas Periprotésicas/cirugía , Estudios Retrospectivos , Reoperación/efectos adversos , Artroplastia de Reemplazo de Cadera/efectos adversos , Prótesis de Cadera/efectos adversos , Complicaciones Posoperatorias/epidemiología , Complicaciones Posoperatorias/etiología , Complicaciones Posoperatorias/cirugía , Fracturas del Fémur/diagnóstico por imagen , Fracturas del Fémur/etiología , Fracturas del Fémur/cirugía
3.
BMC Musculoskelet Disord ; 24(1): 105, 2023 Feb 07.
Artículo en Inglés | MEDLINE | ID: mdl-36750786

RESUMEN

BACKGROUND: The current understanding of glenohumeral joint stability is defined by active restrictions and passive stabilizers including naturally-occurring negative intraarticular pressure. Cadaveric specimens have been used to evaluate the role of intraarticular pressure on joint stability, although, while the shoulder's negative intraarticular pressure is universally acknowledged, it has been inconsistently accounted for. HYPOTHESIS: During continuous, passive humeral abduction, releasing the native intraarticular pressure increases joint translation, and restoring this pressure decreases joint translations. STUDY DESIGN: Descriptive Laboratory Study. METHODS: A validated shoulder testing system was used to passively abduct the humerus in the scapular plane and measure joint translations for seven (n = 7) cadaveric specimens. The pressure within the glenohumeral joint was measured via a 25-gauge needle during passive abduction of the arm, which was released and subsequently restored. During motion, the rotator cuff muscles were loaded using stepper motors in a force feedback loop and electromagnetic sensors were used to continuously measure the position of the humerus and scapula. Joint translation was defined according to the instant center of rotation of the glenohumeral head according to the recommendations by the International Society of Biomechanics. RESULTS: Area under the translation versus abduction angle curve suggests that releasing the pressure within the capsule results in significantly less posterior translation of the glenohumeral head as compared to intact (85-90˚, p < 0.05). Posterior and superior translations were reduced after 70˚ of abduction when the pressure within the joint was restored. CONCLUSION: With our testing system employing a smooth continuous passive motion, we were able to show that releasing intraarticular pressure does not have a major effect on the path of humeral head motion during glenohumeral abduction. However, both violating the capsule and restoring intraarticular pressure after releasing alter glenohumeral translations. Future studies should study the effect of simultaneous external rotation and abduction on the relationship between joint motion and IAP, especially in higher degrees of abduction. CLINICAL RELEVANCE: Thoroughly simulating the glenohumeral joint environment in the cadaveric setting may strengthen the conclusions that can be translated from this setting to the clinic.


Asunto(s)
Articulación del Hombro , Hombro , Humanos , Fenómenos Biomecánicos , Articulación del Hombro/fisiología , Cabeza Humeral , Rango del Movimiento Articular/fisiología , Cadáver
4.
Nat Mater ; 19(1): 102-108, 2020 01.
Artículo en Inglés | MEDLINE | ID: mdl-31844276

RESUMEN

Early insights into the unique structure and properties of native silk suggested that ß-sheet nanocrystallites in silk would degrade prior to melting when subjected to thermal processing. Since then, canonical approaches for fabricating silk-based materials typically involve solution-derived processing methods, which have inherent limitations with respect to silk protein solubility and stability in solution, and time and cost efficiency. Here we report a thermal processing method for the direct solid-state moulding of regenerated silk into bulk 'parts' or devices with tunable mechanical properties. At elevated temperature and pressure, regenerated amorphous silk nanomaterials with ultralow ß-sheet content undergo thermal fusion via molecular rearrangement and self-assembly assisted by bound water to form a robust bulk material that retains biocompatibility, degradability and machinability. This technique reverses presumptions about the limitations of direct thermal processing of silk into a wide range of new material formats and composite materials with tailored properties and functionalities.


Asunto(s)
Materiales Biocompatibles/química , Nanoestructuras/química , Seda/química , Animales , Bombyx , Fuerza Compresiva , Femenino , Fibroínas/química , Calor , Espectroscopía de Resonancia Magnética , Estructura Secundaria de Proteína , Ratas , Ratas Sprague-Dawley , Solubilidad , Espectroscopía Infrarroja por Transformada de Fourier , Estrés Mecánico , Resistencia a la Tracción , Agua/química , Microtomografía por Rayos X
5.
PLoS Pathog ; 13(7): e1006443, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-28704538

RESUMEN

Clostridium difficile (C. difficile) is an anaerobic gram-positive pathogen that is the leading cause of nosocomial bacterial infection globally. C. difficile infection (CDI) typically occurs after ingestion of infectious spores by a patient that has been treated with broad-spectrum antibiotics. While CDI is a toxin-mediated disease, transmission and pathogenesis are dependent on the ability to produce viable spores. These spores must become metabolically active (germinate) in order to cause disease. C. difficile spore germination occurs when spores encounter bile salts and other co-germinants within the small intestine, however, the germination signaling cascade is unclear. Here we describe a signaling role for Ca2+ during C. difficile spore germination and provide direct evidence that intestinal Ca2+ coordinates with bile salts to stimulate germination. Endogenous Ca2+ (released from within the spore) and a putative AAA+ ATPase, encoded by Cd630_32980, are both essential for taurocholate-glycine induced germination in the absence of exogenous Ca2+. However, environmental Ca2+ replaces glycine as a co-germinant and circumvents the need for endogenous Ca2+ fluxes. Cd630_32980 is dispensable for colonization in a murine model of C. difficile infection and ex vivo germination in mouse ileal contents. Calcium-depletion of the ileal contents prevented mutant spore germination and reduced WT spore germination by 90%, indicating that Ca2+ present within the gastrointestinal tract plays a critical role in C. difficile germination, colonization, and pathogenesis. These data provide a biological mechanism that may explain why individuals with inefficient intestinal calcium absorption (e.g., vitamin D deficiency, proton pump inhibitor use) are more prone to CDI and suggest that modulating free intestinal calcium is a potential strategy to curb the incidence of CDI.


Asunto(s)
Ácidos y Sales Biliares/metabolismo , Calcio/metabolismo , Clostridioides difficile/metabolismo , Infecciones por Clostridium/microbiología , Intestino Delgado/microbiología , Esporas Bacterianas/crecimiento & desarrollo , Animales , Proteínas Bacterianas/metabolismo , Señalización del Calcio , Clostridioides difficile/genética , Clostridioides difficile/crecimiento & desarrollo , Infecciones por Clostridium/metabolismo , Humanos , Intestino Delgado/metabolismo , Ratones , Ratones Endogámicos C57BL , Esporas Bacterianas/genética , Esporas Bacterianas/metabolismo
6.
Can J Neurol Sci ; 46(5): 512-517, 2019 09.
Artículo en Inglés | MEDLINE | ID: mdl-31258098

RESUMEN

BACKGROUND: Migraine is a common disorder most typically presenting as headache and often associated with vertigo and motion sickness. It is a genetically complex condition with multiple genes ultimately contributing to the predisposition and development of this episodic neurological disorder. We identified a large American family of 29 individuals of which 17 members suffered from at least one of these disorders, migraine, vertigo, or motion sickness. Many of these individuals suffered from several simultaneously. We hypothesized that vertigo and motion sickness may involve genes that are independent to those directly contributing to migraine susceptibility. METHODS: Genome-wide linkage analysis performed using 400 microsatellite repeat markers spaced at 10 cM throughout the genome. The members of this family were phenotyped for each condition, migraine, vertigo, and motion sickness and analyzed separately. Statistical analysis was performed using two-point and multipoint linkage analysis employing a number of models including autosomal recessive or dominant patterns of inheritance with high and low genetic penetrance. RESULTS: We identified a novel locus for migraine, 9q13-q22 (maximum two-point logarithm of odds [LOD] score-2.51). In addition, there are suggestive LOD scores that localize to different chromosomes for each phenotype; vertigo (chromosome 18, LOD score of 1.82) and motion sickness (chromosome 4, LOD score of 2.09). CONCLUSIONS: Our analysis supports our hypothesis that the migraine-associated vertigo and motion sickness may involve distinct susceptibility genes.


Analyse génétique d'une famille étendue dont les membres souffrent de migraines, de vertiges et du mal des transports. Contexte : La migraine est un trouble courant qui entraîne habituellement des maux de tête et qui est souvent associé à des vertiges et au mal des transports. Il s'agit aussi d'une condition génétique complexe en vertu de laquelle de nombreux gènes contribuent à terme à cette prédisposition et au développement de ce trouble neurologique périodique. À cet égard, nous avons identifié une famille étendue américaine comptant 29 membres. De ce nombre, 17 d'entre eux avaient souffert d'au moins un de ces troubles : des migraines, des vertiges ou le mal des transports. À noter que plusieurs d'entre eux avaient souffert de ces troubles en même temps. Nous avons émis l'hypothèse que les vertiges et le mal des transports pourraient impliquer des gènes qui sont indépendants de ceux contribuant directement à la propension aux migraines. Méthodes : Nous avons effectué une analyse de liaison au moyen de 400 marqueurs microsatellites répétés et espacés à tous les 10 cm au sein de l'ensemble du génome des membres de cette famille. Les membres de cette famille ont été « phénotypés ¼ pour chaque type de trouble (les migraines, les vertiges et le mal des transports) et ont été ensuite analysés de façon séparée. Nous avons effectué une analyse statistique au moyen de l'analyse de liaison multipoint et à deux points, utilisant pour ce faire un certain nombre de modèles, par exemple le modèle autosomique récessif ou des patterns dominants de transmission avec une pénétrance génétique élevée ou faible. Résultats : Nous avons été en mesure d'identifier un nouveau locus dans le cas de la migraine : 9q13-q22 (maximum 2-points ; score au logarithme des probabilités ou LOD : - 2,51). De plus, il est des scores révélateurs au logarithme des probabilités qui permettent de localiser divers chromosomes pour chaque phénotype : vertiges (chromosome 18 ; score au logarithme des probabilités ou LOD : 1,82) et mal des transports (chromosome 4 ; score au logarithme des probabilités ou LOD : 2,09). Conclusions : Notre analyse confirme ainsi notre hypothèse initiale, à savoir que les cas de migraine auxquels sont associés des vertiges et le mal des transports pourraient très bien impliquer différents gènes de susceptibilité.


Asunto(s)
Predisposición Genética a la Enfermedad/genética , Trastornos Migrañosos/genética , Mareo por Movimiento/genética , Vértigo/genética , Adolescente , Niño , Femenino , Ligamiento Genético , Humanos , Masculino , Linaje , Adulto Joven
7.
J Stroke Cerebrovasc Dis ; 28(10): 104287, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31351828

RESUMEN

BACKGROUND: Movement disorders including hemichorea-hemiballism as the initial presentation of an acute ischemic stroke are uncommon. Structures outside of the deep subcortical areas such as the subthalamic nucleus or basal ganglia are rarely involved. CASE REPORT: We report a case of a 72-year-old man with vascular risk factors who presented with acute onset right-sided hemichorea-hemiballism. Metabolic-, infectious-, and toxic-related conditions were ruled out, his EEG was without epileptiform changes. An MRI confirmed an acute ischemic stroke in the parieto-occipital region without any subcortical structures involved. Atrial Fibrillation was later discovered during his hospitalization and was treated appropriately. CONCLUSIONS: Although rare, strokes outside of the subthalamic nucleus can result in hemichorea-hemiballism.


Asunto(s)
Fibrilación Atrial/complicaciones , Infarto Cerebral/etiología , Corea/etiología , Discinesias/etiología , Anciano , Fibrilación Atrial/diagnóstico , Infarto Cerebral/diagnóstico por imagen , Infarto Cerebral/fisiopatología , Corea/diagnóstico , Corea/fisiopatología , Discinesias/diagnóstico , Discinesias/fisiopatología , Humanos , Masculino
8.
J Bacteriol ; 200(16)2018 08 15.
Artículo en Inglés | MEDLINE | ID: mdl-29760211

RESUMEN

Germination of Clostridium difficile spores is a crucial early requirement for colonization of the gastrointestinal tract. Likewise, C. difficile cannot cause disease pathologies unless its spores germinate into metabolically active, toxin-producing cells. Recent advances in our understanding of C. difficile spore germination mechanisms indicate that this process is both complex and unique. This review defines unique aspects of the germination pathways of C. difficile and compares them to those of two other well-studied organisms, Bacillus anthracis and Clostridium perfringensC. difficile germination is unique, as C. difficile does not contain any orthologs of the traditional GerA-type germinant receptor complexes and is the only known sporeformer to require bile salts in order to germinate. While recent advances describing C. difficile germination mechanisms have been made on several fronts, major gaps in our understanding of C. difficile germination signaling remain. This review provides an updated, in-depth summary of advances in understanding of C. difficile germination and potential avenues for the development of therapeutics, and discusses the major discrepancies between current models of germination and areas of ongoing investigation.


Asunto(s)
Clostridioides difficile/fisiología , Esporas Bacterianas/crecimiento & desarrollo , Bacillus anthracis/fisiología , Proteínas Bacterianas/metabolismo , Clostridioides difficile/patogenicidad , Infecciones por Clostridium/tratamiento farmacológico , Clostridium perfringens/fisiología , Humanos , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo
9.
Int J Neurosci ; 126(7): 600-6, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26000935

RESUMEN

AIM: Hereditary spastic paraplegia (HSP) are a genetically and clinically heterogeneous group of disorders. At present, 19 autosomal dominant loci for HSP have been mapped. We ascertained an American family of European descent segregating an autosomal dominant HSP associated with peripheral neuropathy. METHODS: A genome wide scan was performed with 410 microsatellite repeat marker (Weber lab screening set 16) and following linkage and haplotype analysis, fine mapping was performed. Established genes or loci for HSP were excluded by direct sequencing or haplotype analysis. RESULTS: All established loci for HSP were excluded. Fine mapping suggested a locus on chromosome 21q22.3 flanked by markers D21S1411 and D21S1446 with a maximum logarithm of odds score of 2.05 and was supported by haplotype analysis. A number of candidate genes in this region were analyzed and no disease-producing mutations were detected. CONCLUSION: We present the clinical and genetic analysis of an American family with autosomal dominant HSP with axonal sensory motor polyneuropathy mapping to a novel locus on chromosome 21q22.3 designated SPG56.


Asunto(s)
Cromosomas Humanos Par 21/genética , Paraplejía Espástica Hereditaria/genética , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Linaje , Paraplejía Espástica Hereditaria/fisiopatología
10.
Biochemistry ; 54(20): 3197-206, 2015 May 26.
Artículo en Inglés | MEDLINE | ID: mdl-25928210

RESUMEN

Proteins from the enhanced intracellular survival (Eis) family are versatile acetyltransferases that acetylate amines at multiple positions of several aminoglycosides (AGs). Their upregulation confers drug resistance. Homologues of Eis are present in diverse bacteria, including many pathogens. Eis from Mycobacterium tuberculosis (Eis_Mtb) has been well characterized. In this study, we explored the AG specificity and catalytic efficiency of the Eis family protein from Bacillus anthracis (Eis_Ban). Kinetic analysis of specificity and catalytic efficiency of acetylation of six AGs indicates that Eis_Ban displays significant differences from Eis_Mtb in both substrate binding and catalytic efficiency. The number of acetylated amines was also different for several AGs, indicating a distinct regiospecificity of Eis_Ban. Furthermore, most recently identified inhibitors of Eis_Mtb did not inhibit Eis_Ban, underscoring the differences between these two enzymes. To explain these differences, we determined an Eis_Ban crystal structure. The comparison of the crystal structures of Eis_Ban and Eis_Mtb demonstrates that critical residues lining their respective substrate binding pockets differ substantially, explaining their distinct specificities. Our results suggest that acetyltransferases of the Eis family evolved divergently to garner distinct specificities while conserving catalytic efficiency, possibly to counter distinct chemical challenges. The unique specificity features of these enzymes can be utilized as tools for developing AGs with novel modifications and help guide specific AG treatments to avoid Eis-mediated resistance.


Asunto(s)
Acetiltransferasas/química , Bacillus anthracis/enzimología , Proteínas Bacterianas/química , Acetilación , Acetiltransferasas/antagonistas & inhibidores , Acetiltransferasas/metabolismo , Secuencia de Aminoácidos , Antibacterianos/farmacología , Bacillus anthracis/efectos de los fármacos , Proteínas Bacterianas/antagonistas & inhibidores , Proteínas Bacterianas/metabolismo , Dominio Catalítico , Cristalografía por Rayos X , Farmacorresistencia Bacteriana , Concentración 50 Inhibidora , Cinética , Pruebas de Sensibilidad Microbiana , Modelos Moleculares , Datos de Secuencia Molecular , Procesamiento Proteico-Postraduccional , Estructura Secundaria de Proteína
12.
Anaerobe ; 33: 64-70, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25681667

RESUMEN

Over the past two decades, Clostridium difficile infections have been increasing in both number and severity throughout the world. As with other spore forming bacteria, germination is a vital step in the life cycle of this pathogen. Studies have examined differences in sporulation and toxin production among a number of C. difficile clinical isolates; however, few have examined differences in germination and the relationship between this phenotype and disease severity. Here, over 100 C. difficile isolates from the University of Michigan Health System were examined for overall germination in response to various combinations of known germinants (taurocholate) and co-germinants (glycine and histidine). Significant variation was observed among isolates under all conditions tested. Isolates representing ribotype 014-020, which was the most frequently isolated ribotype at our hospital, exhibited increased germination in the presence of taurocholate and glycine when compared to isolates representing other ribotypes. Interestingly, isolates that caused severe disease exhibited significantly lower germination in response to minimal germination conditions (taurocholate only), indicating increased control over germination in these isolates. These data provide a broad picture of C. difficile isolate germination and indicate a role for precise control of germination in disease severity.


Asunto(s)
Clostridioides difficile/fisiología , Enterocolitis Seudomembranosa/microbiología , Esporas Bacterianas , Clostridioides difficile/aislamiento & purificación , Enterocolitis Seudomembranosa/diagnóstico , Humanos , Índice de Severidad de la Enfermedad
13.
J Am Chem Soc ; 136(4): 1579-86, 2014 Jan 29.
Artículo en Inglés | MEDLINE | ID: mdl-24401083

RESUMEN

Siderophores are high-affinity iron chelators produced by microorganisms and frequently contribute to the virulence of human pathogens. Targeted inhibition of the biosynthesis of siderophores staphyloferrin B of Staphylococcus aureus and petrobactin of Bacillus anthracis hold considerable potential as a single or combined treatment for methicillin-resistant S. aureus (MRSA) and anthrax infection, respectively. The biosynthetic pathways for both siderophores involve a nonribosomal peptide synthetase independent siderophore (NIS) synthetase, including SbnE in staphyloferrin B and AsbA in petrobactin. In this study, we developed a biochemical assay specific for NIS synthetases to screen for inhibitors of SbnE and AsbA against a library of marine microbial-derived natural product extracts (NPEs). Analysis of the NPE derived from Streptomyces tempisquensis led to the isolation of the novel antibiotics baulamycins A (BmcA, 6) and B (BmcB, 7). BmcA and BmcB displayed in vitro activity with IC50 values of 4.8 µM and 19 µM against SbnE and 180 µM and 200 µM against AsbA, respectively. Kinetic analysis showed that the compounds function as reversible competitive enzyme inhibitors. Liquid culture studies with S. aureus , B. anthracis , E. coli , and several other bacterial pathogens demonstrated the capacity of these natural products to penetrate bacterial barriers and inhibit growth of both Gram-positive and Gram-negative species. These studies provide proof-of-concept that natural product inhibitors targeting siderophore virulence factors can provide access to novel broad-spectrum antibiotics, which may serve as important leads for the development of potent anti-infective agents.


Asunto(s)
Antibacterianos/farmacología , Bacillus anthracis/efectos de los fármacos , Productos Biológicos/farmacología , Daunorrubicina/análogos & derivados , Escherichia coli/efectos de los fármacos , Sideróforos/antagonistas & inhibidores , Staphylococcus aureus/efectos de los fármacos , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Bacillus anthracis/química , Bacillus anthracis/metabolismo , Productos Biológicos/química , Productos Biológicos/aislamiento & purificación , Daunorrubicina/síntesis química , Daunorrubicina/química , Daunorrubicina/farmacología , Relación Dosis-Respuesta a Droga , Ensayos Analíticos de Alto Rendimiento , Pruebas de Sensibilidad Microbiana , Conformación Molecular , Sideróforos/biosíntesis , Staphylococcus aureus/química , Staphylococcus aureus/metabolismo , Relación Estructura-Actividad
14.
ACS Biomater Sci Eng ; 10(4): 2607-2615, 2024 04 08.
Artículo en Inglés | MEDLINE | ID: mdl-38478959

RESUMEN

Conventional thinking when designing biodegradable materials and devices is to tune the intrinsic properties and morphological features of the material to regulate their degradation rate, modulating traditional factors such as molecular weight and crystallinity. Since regenerated silk protein can be directly thermoplastically molded to generate robust dense silk plastic-like materials, this approach afforded a new tool to control silk degradation by enabling the mixing of a silk-degrading protease into bulk silk material prior to thermoplastic processing. Here we demonstrate the preparation of these silk-based devices with embedded silk-degrading protease to modulate the degradation based on the internal presence of the enzyme to support silk degradation, as opposed to the traditional surface degradation for silk materials. The degradability of these silk devices with and without embedded protease XIV was assessed both in vitro and in vivo. Ultimately, this new process approach provides direct control of the degradation lifetime of the devices, empowered through internal digestion via water-activated proteases entrained and stabilized during the thermoplastic process.


Asunto(s)
Materiales Biocompatibles , Seda , Péptido Hidrolasas , Agua
15.
J Biol Chem ; 287(19): 16058-72, 2012 May 04.
Artículo en Inglés | MEDLINE | ID: mdl-22408253

RESUMEN

Petrobactin, a mixed catechol-carboxylate siderophore, is required for full virulence of Bacillus anthracis, the causative agent of anthrax. The asbABCDEF operon encodes the biosynthetic machinery for this secondary metabolite. Here, we show that the function of five gene products encoded by the asb operon is necessary and sufficient for conversion of endogenous precursors to petrobactin using an in vitro system. In this pathway, the siderophore synthetase AsbB catalyzes formation of amide bonds crucial for petrobactin assembly through use of biosynthetic intermediates, as opposed to primary metabolites, as carboxylate donors. In solving the crystal structure of the B. anthracis siderophore biosynthesis protein B (AsbB), we disclose a three-dimensional model of a nonribosomal peptide synthetase-independent siderophore (NIS) synthetase. Structural characteristics provide new insight into how this bifunctional condensing enzyme can bind and adenylate multiple citrate-containing substrates followed by incorporation of both natural and unnatural polyamine nucleophiles. This activity enables formation of multiple end-stage products leading to final assembly of petrobactin. Subsequent enzymatic assays with the nonribosomal peptide synthetase-like AsbC, AsbD, and AsbE polypeptides show that the alternative products of AsbB are further converted to petrobactin, verifying previously proposed convergent routes to formation of this siderophore. These studies identify potential therapeutic targets to halt deadly infections caused by B. anthracis and other pathogenic bacteria and suggest new avenues for the chemoenzymatic synthesis of novel compounds.


Asunto(s)
Bacillus anthracis/metabolismo , Proteínas Bacterianas/metabolismo , Benzamidas/metabolismo , Vías Biosintéticas , Ligasas de Carbono-Nitrógeno/metabolismo , Ligasas/metabolismo , Secuencia de Aminoácidos , Bacillus anthracis/genética , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Benzamidas/química , Biocatálisis , Ligasas de Carbono-Nitrógeno/química , Ligasas de Carbono-Nitrógeno/genética , Cristalografía por Rayos X , Ligasas/química , Ligasas/genética , Modelos Químicos , Modelos Moleculares , Datos de Secuencia Molecular , Estructura Molecular , Poliaminas/química , Poliaminas/metabolismo , Unión Proteica , Estructura Terciaria de Proteína , Homología de Secuencia de Aminoácido , Sideróforos/química , Sideróforos/metabolismo , Especificidad por Sustrato
16.
Mol Microbiol ; 84(2): 370-82, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22429808

RESUMEN

In Bacillus anthracis the siderophore petrobactin is vital for iron acquisition and virulence. The petrobactin-binding receptor FpuA is required for these processes. Here additional components of petrobactin reacquisition are described. To identify these proteins, mutants of candidate permease and ATPase genes were generated allowing for characterization of multiple petrobactin ATP-binding cassette (ABC)-import systems. Either of two distinct permeases, FpuB or FatCD, is required for iron acquisition and play redundant roles in petrobactin transport. A mutant strain lacking both permeases, ΔfpuBΔfatCD, was incapable of using petrobactin as an iron source and exhibited attenuated virulence in a murine model of inhalational anthrax infection. ATPase mutants were generated in either of the permease mutant backgrounds to identify the ATPase(s) interacting with each individual permease channel. Mutants lacking the FpuB permease and FatE ATPase (ΔfpuBΔfatE) and a mutant lacking the distinct ATPases FpuC and FpuD generated in the ΔfatCD background (ΔfatCDΔfpuCΔfpuD) displayed phenotypic characteristics of a mutant deficient in petrobactin import. A mutant lacking all three of the identified ATPases (ΔfatEΔfpuCΔfpuD) exhibited the same growth defect in iron-depleted conditions. Taken together, these results provide the first description of the permease and ATPase proteins required for the import of petrobactin in B. anthracis.


Asunto(s)
Transportadoras de Casetes de Unión a ATP/metabolismo , Bacillus anthracis/metabolismo , Bacillus anthracis/patogenicidad , Benzamidas/metabolismo , Factores de Virulencia/metabolismo , Transportadoras de Casetes de Unión a ATP/genética , Animales , Carbunco/microbiología , Carbunco/patología , Bacillus anthracis/genética , Modelos Animales de Enfermedad , Eliminación de Gen , Hierro/metabolismo , Ratones , Modelos Biológicos , Mutagénesis , Análisis de Supervivencia , Virulencia , Factores de Virulencia/genética
17.
Anaerobe ; 24: 109-16, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-23608205

RESUMEN

Since 2000, Clostridium difficile isolates of ribotype 027 have been linked to outbreaks in North America and Europe and also an increased rate of colectomy and death among infected individuals. It has been proposed that enhanced sporulation and toxin production were associated with this apparent increase in virulence of 027 isolates. Since only a limited number of isolates have been examined, the relationship of these phenotypes to a specific ribotype, and as well as to clinical disease severity, remains controversial. 106 recent clinical isolates from the University of Michigan Health System were characterized for the ability to sporulate, produce viable spores, grow in rich media, and produce toxins in vitro. Significant variation was observed between isolates for each of these phenotypes. Isolates of ribotype 027 produced higher levels of toxin and exhibited slower growth compared to other ribotypes. Importantly, increased spore production did appear to be relevant to severe C. difficile infection, as determined by available clinical meta-data. These data provide the first significant difference between isolates from severe vs. less severe disease based on an in vitro C. difficile phenotype and suggest that clinical outcome is better predicted by bacterial attributes other than ribotype.


Asunto(s)
Toxinas Bacterianas/metabolismo , Clostridioides difficile/genética , Clostridioides difficile/patogenicidad , Infecciones por Clostridium/microbiología , Infecciones por Clostridium/patología , Ribotipificación , Toxinas Bacterianas/genética , Clostridioides difficile/clasificación , Clostridioides difficile/aislamiento & purificación , Genotipo , Humanos , Michigan , Fenotipo , Índice de Severidad de la Enfermedad , Esporas Bacterianas/crecimiento & desarrollo , Virulencia
18.
Arthrosc Tech ; 12(4): e531-e536, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-37138697

RESUMEN

Advances in regional anesthesia techniques for knee surgery have led to drastic improvements in postoperative pain control and have reduced reliance on perioperative opioid analgesics. The infiltration between the popliteal artery and capsule of the knee (IPACK) block has been a useful tool for providing posterior knee analgesia as an adjuvant to traditional femoral or adductor canal blocks in knee surgery. We present a simple and reproducible technique for the arthroscopic administration of this block.

19.
JBJS Case Connect ; 13(4)2023 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-38011320

RESUMEN

CASE: A 12-year-old adolescent boy presented with a proximal fibula fracture and lateral ankle dislocation consistent with a Maisonneuve fracture (MF) associated with a transsyndesmotic ankle dislocation. The dislocation was reduced under conscious sedation in the emergency department. Postreduction imaging studies demonstrated a Tillaux fracture. The patient underwent surgical stabilization of the Tillaux fragment and of the distal tibiofibular syndesmosis. At the 26-month follow-up, the patient remained active without restrictions. CONCLUSION: Operative treatment of a concurrent MF, Tillaux fracture with lateral ankle dislocation, or a pediatric "logsplitter" injury resulted in satisfactory alignment and function of the ankle joint.


Asunto(s)
Fracturas de Tobillo , Fracturas de Peroné , Luxaciones Articulares , Fracturas de la Tibia , Masculino , Adolescente , Humanos , Niño , Articulación del Tobillo/diagnóstico por imagen , Articulación del Tobillo/cirugía , Tobillo , Fracturas de Tobillo/diagnóstico por imagen , Fracturas de Tobillo/cirugía , Fijación Interna de Fracturas/métodos , Luxaciones Articulares/diagnóstico por imagen , Luxaciones Articulares/cirugía
20.
Hip Int ; 33(4): 771-778, 2023 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-35836328

RESUMEN

BACKGROUND: Despite the fact that patients with chronic liver disease (CLD) are at increased risk of complications after a fracture of the hip, there remains little information about the risk factors for acute postoperative complications and their overall outcome.The aim of this study was to describe inpatient postoperative complications and identify predictors of postoperative morbidity. METHODS: Patients with CLD who had been treated for a fracture of the hip between April 2005 and August 2019 were identified from a retrospective search of an intramural trauma registry based in the Northeastern United States. Medical records were reviewed for baseline demographics, preoperative laboratory investigations, and outcomes. RESULTS: The trauma registry contained 110 patients with CLD who had undergone surgery for a fracture of the hip. Of these, patients with a platelet-count of ⩽100,000/µL were 3.81 (95% CI, 1.59-9.12) times more likely to receive a transfusion than those with a platelet-count of >100,000/µL. Those with a Model for End-stage Liver Disease (MELD) score of >9 were 5.54 (2.33-13.16) times more likely to receive a transfusion and 3.97 (1.06-14.81) times more likely to develop postoperative delirium than those with a MELD score of ⩽9.Of patients without chronic kidney disease, those with a creatinine of ⩾1.2 mg/dL were 6.80 (1.79-25.87) times more likely to develop acute renal failure (ARF) than those with a creatinine of <1.2 mg/dL. In a multivariable model, as MELD score was increased, the odds of developing a composite postoperative complication, which included transfusion, ARF, delirium, or deep wound infection, were 1.29 (1.01-1.66). Other tools used to assess surgical risks, Charlson Comorbidity Index, Elixhauser, and American Society of Anesthesiologist scores, were not predictive. CONCLUSIONS: Patients with CLD who undergo surgery for a hip fracture have a high rate of postoperative complications which can be predicted by the preoperative laboratory investigations identified in this study and MELD scores, but not by other common comorbidity indices.


Asunto(s)
Artroplastia de Reemplazo de Cadera , Enfermedad Hepática en Estado Terminal , Fracturas de Cadera , Hepatopatías , Humanos , Enfermedad Hepática en Estado Terminal/complicaciones , Enfermedad Hepática en Estado Terminal/cirugía , Estudios Retrospectivos , Creatinina , Artroplastia de Reemplazo de Cadera/efectos adversos , Índice de Severidad de la Enfermedad , Fracturas de Cadera/etiología , Factores de Riesgo , Hepatopatías/complicaciones , Hepatopatías/cirugía , Complicaciones Posoperatorias/epidemiología , Complicaciones Posoperatorias/etiología , Complicaciones Posoperatorias/cirugía , Morbilidad
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