Asunto(s)
Selectina E , Leucemia Mieloide Aguda , Humanos , Tirosina Quinasa 3 Similar a fms , Leucemia Mieloide Aguda/tratamiento farmacológico , Ligandos , Mutación , Inhibidores de Proteínas Quinasas/farmacología , Receptores CXCR4/genética , Transducción de Señal , Sistema de Señalización de MAP QuinasasRESUMEN
ABSTRACT: A 50-year-old man, with a history of extensive sun exposure and multiple previous non-melanoma skin cancers, presented with an asymptomatic 8-× 10-millimeter scaly, skin-colored papule on his right shoulder. Subsequent biopsy and excision revealed epidermal hyperplasia containing large atypical basaloid cells with pagetoid spread. Immunoperoxidase staining for cytokeratin-20 demonstrated a focal perinuclear dot-like pattern, and after excluding other in situ entities, a diagnosis of Merkel cell carcinoma In Situ (MCCIS) was rendered. MCCIS is a very rare entity. Although approximately 18% of Merkel cell carcinomas have epidermal involvement, currently only 17 cases of MCCIS have been reported, of which only 7 had no associated neoplasm. Previously, MCCIS was considered a serendipitous or incidental finding, as most cases co-existed with squamous cell carcinoma in situ. This case is unique in that it was not associated with a squamous lesion, and in addition, the pagetoid spread was unusual and has only occasionally been described. As such, MCCIS should be added to list of in situ epidermal lesions with pagetoid spread.
Asunto(s)
Carcinoma de Células de Merkel/diagnóstico , Neoplasias Cutáneas/diagnóstico , Carcinoma de Células de Merkel/patología , Carcinoma de Células de Merkel/cirugía , Diagnóstico Diferencial , Humanos , Masculino , Persona de Mediana Edad , Neoplasias Cutáneas/patología , Neoplasias Cutáneas/cirugíaAsunto(s)
Anemia de Células Falciformes/metabolismo , Capacidad de Concentración Renal , Enfermedades Renales/etiología , Receptores de Angiotensina/metabolismo , Adulto , Anemia de Células Falciformes/complicaciones , Bloqueadores del Receptor Tipo 1 de Angiotensina II/uso terapéutico , Animales , Captopril/farmacología , Niño , Progresión de la Enfermedad , Humanos , Losartán/farmacología , Ratones , Sustancias Protectoras , Receptor de Angiotensina Tipo 2/agonistas , Transducción de SeñalRESUMEN
Life-long hematopoietic demands are met by a pool of hematopoietic stem cells (HSC) with self-renewal and multipotential differentiation ability. Humoral and paracrine signals from the bone marrow (BM) hematopoietic microenvironment control HSC activity. Cell-to-cell communication through connexin (Cx) containing gap junctions (GJs) allows pluricellular coordination and synchronization through transfer of small molecules with messenger activity. Hematopoietic and surrounding nonhematopoietic cells communicate each other through GJs, which regulate fetal and postnatal HSC content and function in hematopoietic tissues. Traffic of HSC between peripheral blood and BM is also dependent on Cx proteins. Cx mutations are associated with human disease and hematopoietic dysfunction and Cx signaling may represent a target for therapeutic intervention. In this review, we illustrate and highlight the importance of Cxs in the regulation of hematopoietic homeostasis under normal and pathological conditions.
Asunto(s)
Conexinas/metabolismo , Células Madre Hematopoyéticas/metabolismo , Tejido Linfoide/metabolismo , Comunicación Paracrina/fisiología , Transducción de Señal/fisiología , Nicho de Células Madre/fisiología , Animales , Células de la Médula Ósea/citología , Células de la Médula Ósea/metabolismo , Uniones Comunicantes/metabolismo , Células Madre Hematopoyéticas/citología , Humanos , Tejido Linfoide/citologíaRESUMEN
The aim of the present study was to assess the bioequivalence of two cephalexin tablet formulations available in the Brazilian market (product A as reference formulation and product B as test formulation). Dissolution efficiency (DE%) was calculated for both formulations to evaluate their
O objetivo do presente estudo foi avaliar a bioequivalência de duas formulações de cefalexina disponíveis no mercado brasileiro (produto A como formulação referência e produto B como formulação teste). A eficiência de dissolução (DE%) foi calculada para ambas as formulações para avaliar suas características biofarmacêuticas. O estudo de bioequivalência oral foi realizado em vinte e quatro voluntários sadios utilizando um desenho cruzado. Uma dose oral única (comprimido contendo 500 mg de cefalexina) de cada produto foi administrada com um período de
Asunto(s)
Humanos , Cefalexina/farmacocinética , Cefalexina/farmacología , Cromatografía Líquida de Alta Presión , Equivalencia Terapéutica , Orina/químicaRESUMEN
Patients with organ failure of vascular origin have increased circulating haematopoietic stem cells and progenitors (HSC/P). Plasma levels of angiotensin II (Ang-II), are commonly increased in vasculopathies. Hyperangiotensinemia results in activation of a very distinct Ang-II receptor set, Rho family GTPase members, and actin in bone marrow endothelial cells (BMEC) and HSC/P, which results in decreased membrane integrin activation in both BMEC and HSC/P, and in HSC/P de-adhesion and mobilization. The Ang-II effect can be reversed pharmacologically and genetically by inhibiting Ang-II production or signalling through BMEC AT2R, HSCP Ang-II receptor type 1 (AT1R)/AT2R or HSC/P RhoA, but not by interfering with other vascular tone mediators. Hyperangiotensinemia and high counts of circulating HSC/P seen in sickle cell disease (SCD) as a result of vascular damage, is significantly decreased by Ang-II inhibitors. Our data define for the first time the role of Ang-II HSC/P traffic regulation and redefine the haematopoietic consequences of anti-angiotensin therapy in SCD.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Angiotensina II/metabolismo , Citoesqueleto/metabolismo , Células Madre Hematopoyéticas/citología , Células Madre/citología , Enfermedades Vasculares/patología , Actinas/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Anemia de Células Falciformes/metabolismo , Animales , Células de la Médula Ósea/citología , Adhesión Celular , Membrana Celular/metabolismo , Cruzamientos Genéticos , Células Endoteliales/citología , Hematopoyesis , Humanos , Integrina beta1/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Microscopía Confocal , Óxido Nítrico/química , Transducción de SeñalRESUMEN
In the bone marrow (BM), hematopoietic progenitors (HPs) reside in specific anatomical niches near osteoblasts (Obs), macrophages (MΦs), and other cells forming the BM microenvironment. A connection between immunosurveillance and traffic of HP has been demonstrated, but the regulatory signals that instruct the immune regulation of HP circulation are unknown. We discovered that the BM microenvironment deficiency of p62, an autophagy regulator and signal organizer, results in loss of autophagic repression of macrophage contact-dependent activation of Ob NF-κB signaling. Consequently, Ob p62-deficient mice lose bone, Ob Ccl4 expression, and HP chemotaxis toward Cxcl12, resulting in egress of short-term hematopoietic stem cells and myeloid progenitors. Finally, Ccl4 expression and myeloid progenitor egress are reversed by deficiency of the p62 PB1-binding partner Nbr1. A functional "MΦ-Ob niche" is required for myeloid progenitor/short-term stem cell retention, in which Ob p62 is required to maintain NF-κB signaling repression, osteogenesis, and BM progenitor retention.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas de Choque Térmico/metabolismo , Células Madre Hematopoyéticas/metabolismo , Macrófagos/metabolismo , Osteoblastos/metabolismo , Osteogénesis , Transducción de Señal , Nicho de Células Madre , Proteínas Adaptadoras Transductoras de Señales/genética , Animales , Autofagia , Quimiocina CCL4/metabolismo , Proteínas de Choque Térmico/genética , Células Madre Hematopoyéticas/citología , Quinasa I-kappa B/metabolismo , Péptidos y Proteínas de Señalización Intracelular , Macrófagos/citología , Ratones , FN-kappa B/metabolismo , Osteoblastos/citología , Proteínas/metabolismo , Proteína Sequestosoma-1RESUMEN
We report 2 patients with typical clinical findings of circumscribed morphea who on histopathologic examinations had histiocytes ("floating sign") surrounding individual collagen fibers in the dermis in addition to the key histologic findings of morphea. To our knowledge, there are no previous reports in the medical literature of such a phenomenon. Histopathological findings in idiopathic morphea and morphea-like conditions are reviewed.
Asunto(s)
Histiocitos/patología , Esclerodermia Localizada/patología , Adulto , Femenino , HumanosRESUMEN
The Rho family of guanosine triphosphatases (GTPases) is composed of members of the Ras superfamily of proteins. They are GTP-bound molecules with a modest intrinsic GTPase activity that can be accelerated upon activation/localization of specialized guanine nucleotide exchange factors. Members of this family act as molecular switches and are required for coordinated cytoskeletal rearrangements that are crucial in a set of specialized functions of mammalian stem cells. These functions include self-renewal, adhesion, and migration. Mouse gene-targeting studies have provided convincing evidence of the indispensable and dispensable roles of individual members of the Rho GTPase family and the putative upstream and downstream mediators in stem cell-specific functions. The role of Rho GTPases and related signaling pathways previously seen in other cell types and organisms have been confirmed in mammalian hematopoietic stem cells (HSCs), and new signaling pathways and unexpected functions unique to HSCs have been identified and dissected. This review summarizes our current understanding of the role of Rho family of GTPases on HSC and progenitor activity through cytoskeleton-mediated signaling pathways, providing insight about relevant signaling pathways that regulate mammalian stem cell self-renewal, adhesion, and migration.
Asunto(s)
Citoesqueleto/fisiología , Células Madre Hematopoyéticas/fisiología , Proteínas de Unión al GTP rho/metabolismo , Animales , Hematopoyesis/fisiología , HumanosRESUMEN
Despite the introduction of tyrosine kinase inhibitor therapy, the prognosis for p190-BCR-ABL(+) acute lymphoblastic leukemia remains poor. In the present study, we present the cellular and molecular roles of the Rho GTPase guanine nucleotide exchange factor Vav in lymphoid leukemogenesis and explore the roles of Vav proteins in BCR-ABL-dependent signaling. We show that genetic deficiency of the guanine nucleotide exchange factor Vav3 delays leukemogenesis by p190-BCR-ABL and phenocopies the effect of Rac2 deficiency, a downstream effector of Vav3. Compensatory up-regulation of expression and activation of Vav3 in Vav1/Vav2-deficient B-cell progenitors increases the transformation ability of p190-BCR-ABL. Vav3 deficiency induces apoptosis of murine and human leukemic lymphoid progenitors, decreases the activation of Rho GTPase family members and p21-activated kinase, and is associated with increased Bad phosphorylation and up-regulation of Bax, Bak, and Bik. Finally, Vav3 activation only partly depends on ABL TK activity, and Vav3 deficiency collaborates with tyrosine kinase inhibitors to inhibit CrkL activation and impair leukemogenesis in vitro and in vivo. We conclude that Vav3 represents a novel specific molecular leukemic effector for multitarget therapy in p190-BCR-ABL-expressing acute lymphoblastic leukemia.
Asunto(s)
Linfocitos B/patología , Transformación Celular Neoplásica/patología , Proteínas de Fusión bcr-abl/metabolismo , Células Progenitoras Linfoides/patología , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Proteínas Proto-Oncogénicas c-vav/fisiología , Animales , Linfocitos B/metabolismo , Transformación Celular Neoplásica/metabolismo , Células Cultivadas , Femenino , Sangre Fetal/citología , Sangre Fetal/metabolismo , Humanos , Células Progenitoras Linfoides/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Fosforilación , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Tasa de Supervivencia , Ensayo de Tumor de Célula Madre , Proteínas de Unión al GTP rac/fisiología , Proteína RCA2 de Unión a GTPRESUMEN
BACKGROUND: Squamous cell carcinoma in situ (SCCIS) is thought to be a precursor to squamous cell carcinoma. It should be treated before invasive cancer develops, especially in transplant recipients, who may develop more aggressive skin cancers. Treatment can involve surgical and nonsurgical methods. OBJECTIVE To review the evidence available in the English medical literature for different treatment options of SCCIS on nongenital skin and evaluate the efficacy of each option. METHODS AND MATERIALS: A Pubmed search of articles describing the treatment of SCCIS was conducted. Keywords were "treatment," "Bowen's disease," and "squamous cell carcinoma in situ." Articles describing the use of surgical excision, curettage and electrodesiccation, cryotherapy, 5-fluorouracil, imiquimod, radiation, photodynamic therapy, lasers, and rarer methods were reviewed. RESULTS: No single treatment can be said to be superior for any one situation. Most studies are small, limiting the power of each. Further studies are needed to clarify optimal treatment protocols for nonsurgical methods such as cryotherapy, photodynamic therapy, and topical chemotherapy. CONCLUSION: There are many methods available to treat SCCIS. Physicians should consider each patient's situation while keeping in mind that treatment protocols have not been fully defined for most options. The authors have indicated no significant interest with commercial supporters.
Asunto(s)
Enfermedad de Bowen/terapia , Carcinoma in Situ/terapia , Carcinoma de Células Escamosas/terapia , Neoplasias Cutáneas/terapia , HumanosRESUMEN
BACKGROUND: Melanoma is a life-threatening malignancy. Surgery is the primary management for melanoma, and management guidelines have evolved gradually over a century from radical surgery with lymph node dissection to conservative margin surgery. There are specific rationales and problems with Mohs micrographic (MMS) surgery for managing melanoma. OBJECTIVE: To review the literature for the surgical management of melanoma and to understand where MMS fits in this spectrum of management options. CONCLUSIONS: MMS should be considered as an option for melanoma surgery, especially when the tumor is found in photodamaged skin. Further randomized prospective clinical trials are needed to select the best therapeutic approach for the treatment of melanoma. Until then, careful margin control is the key for successful tumor removal whether it is standard excision, staged excision, or MMS.
Asunto(s)
Melanoma/cirugía , Cirugía de Mohs , Neoplasias Cutáneas/cirugía , Humanos , Melanoma/patología , Guías de Práctica Clínica como Asunto , Neoplasias Cutáneas/patologíaRESUMEN
BACKGROUND: Immunohistochemistry (IHC) applied to Mohs micrographic surgery (MMS) is time consuming and labor intensive, and the variability of staining quality has prevented its widespread use in clinical practice. OBJECTIVE: To investigate the readability of immunostains processed by a novel automated 16-minute technique used for evaluation of frozen sections prepared during MMS for melanoma. METHODS: A rapid automated instrument that performs MART-1 (melanoma antigen recognized by T cells) immunostains in 16 minutes was used to stain frozen sections and was compared with MART-1 stains of paraffin (permanent) sections, hematoxylin-eosin (H&E) stains of frozen and permanent sections from the positive or negative control specimens of the Mohs layers for melanoma. A total of 480 interpretations from 48 sections (4 types of stains for each specimen, 12 specimens read by 10 interpreters) were analyzed via blinded examination by 5 dermatopathologists and 5 Mohs surgeons at two institutions. A scoring system was used to assess the readability of each slide. Analysis of variance was used for statistical analysis. RESULTS: In terms of clarity of interpreting melanoma sections, the 16-minute MART-1 IHC of frozen sections is equivalent to the standard MART-1 of permanent sections. The 16-minute MART-1 sections are also significantly easier to interpret than permanent sections stained with H&E for both the dermatopathologists and Mohs surgeons (P < .05). LIMITATIONS: The study represents data collected from only two institutions in the United States. CONCLUSION: The rapid-stained frozen IHC sections are significantly easier to interpret than the "gold standard" permanent sections stained with H&E. This technology facilitates the rapid interpretation of melanoma in frozen sections.
Asunto(s)
Antígenos Específicos del Melanoma/análisis , Melanoma/cirugía , Cirugía de Mohs/métodos , Neoplasias Cutáneas/cirugía , Coloración y Etiquetado/métodos , Automatización , Biopsia con Aguja , Eosina Amarillenta-(YS) , Estudios de Evaluación como Asunto , Femenino , Secciones por Congelación/métodos , Hematoxilina , Humanos , Inmunohistoquímica , Cuidados Intraoperatorios/métodos , Masculino , Melanoma/patología , Muestreo , Sensibilidad y Especificidad , Neoplasias Cutáneas/patología , Factores de Tiempo , Estados UnidosRESUMEN
BACKGROUND: Colonization with methicillin-resistant Staphylococcus aureus (MRSA) places patients at risk for postoperative MRSA wound infections. OBJECTIVE: To determine the effect of a decontamination and prophylaxis protocol on postoperative MRSA wound infections in patients with nasal MRSA. METHODS & MATERIALS: Wound cultures over a 23-month period were reviewed before and 11 months after implementation of a screening and decontamination protocol. After preoperative MRSA screening with nasal swabs, carriers were instructed to use intranasal mupirocin for 5 to 7 days before surgery and 5 to 7 days of trimethoprim-sulfamethoxazole starting the day before surgery. RESULTS: During the 23 months before prescreening evaluation, we performed 3,633 Mohs surgical cases, and 12 postoperative MRSA wound infections (0.3%) occurred. Subsequently, 963 patients underwent screening for MRSA, and 23 MRSA carriers were identified (2.4%). Of the 22 who underwent the decontamination and treatment protocol, none developed postoperative wound infections. One MRSA carrier did not receive preoperative treatment and subsequently developed a MRSA wound infection. There were no other MRSA infections. CONCLUSION: Preoperative MRSA screening and implementation of a decontamination protocol appears to decrease postoperative MRSA wound infections after Mohs surgery. Although an interesting observation, controlled studies of clinical and cost effectiveness are required before general implementation. The authors have indicated no significant interest with commercial supporters.
Asunto(s)
Control de Infecciones/métodos , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Cirugía de Mohs/efectos adversos , Cuidados Preoperatorios/métodos , Infecciones Estafilocócicas/prevención & control , Infección de la Herida Quirúrgica/microbiología , Infección de la Herida Quirúrgica/prevención & control , Administración Tópica , Antibacterianos/administración & dosificación , Profilaxis Antibiótica , Portador Sano/diagnóstico , Descontaminación/métodos , Hospitales de Veteranos , Humanos , Tamizaje Masivo/métodos , Mupirocina/administración & dosificación , Estudios Retrospectivos , Infecciones Estafilocócicas/diagnóstico , Infecciones Estafilocócicas/microbiologíaRESUMEN
PURPOSE: Modulators of angiogenesis typically work in an orchestrated manner. The authors examined the interaction between insulinlike growth factor (IGF)-1, vascular endothelial growth factor (VEGF), and stromal derived factor (SDF)-1 in vivo and in vitro using angiogenesis models. METHODS: The angiogenic effect of SDF-1, alone or in combination with IGF-1 and VEGF, was assessed in human lung microvascular endothelial cells using capillary tube formation and thymidine incorporation. Immunohistochemical analysis for CD31, SDF-1, and CXCR4 was performed on mouse eyes 2 weeks after the initiation of laser rupture of Bruch's membrane, a choroidal neovascularization (CNV) model. CXCR4 antagonist and CXCR4 blocking antibody were tested on inhibition of CNV lesion size in this model. Real-time PCR was used to determine mRNA levels for SDF-1, VEGF, IGF-1, and their cognate receptors in the retinal pigment epithelium/choroid complex of mice that underwent this CNV model. RESULTS: IGF-1 and VEGF demonstrated an additive effect on SDF-1-induced in vitro angiogenesis. CXCR4 immunoreactivity was present in both normal and laser-injured mice at the laser burn site and at the ganglion cell layer, the anterior portion of the inner nuclear layer, photoreceptors, and choroidal stroma. SDF-1 was observed in identical locations but was not seen in photoreceptors. mRNA levels for SDF-1, VEGF, and IGF-1 and their receptors were increased after laser injury. CXCR4-neutralizing antibody reduced neovascularization when injected subretinally but not intraperitoneally or intravitreally. CONCLUSIONS: The potent proangiogenic factors IGF-1 and VEGF both stimulate SDF-1-induced angiogenesis. Local inhibition of CXCR4 is required for an antiangiogenic effect in CNV lesions.
Asunto(s)
Proteínas Angiogénicas/farmacología , Neovascularización Coroidal/metabolismo , Endotelio Vascular/efectos de los fármacos , Factor I del Crecimiento Similar a la Insulina/farmacología , Comunicación Paracrina/fisiología , Receptores CXCR4/metabolismo , Factor A de Crecimiento Endotelial Vascular/farmacología , Animales , Quimiocina CXCL12/genética , Quimiocina CXCL12/farmacología , Neovascularización Coroidal/patología , Modelos Animales de Enfermedad , Endotelio Vascular/patología , Humanos , Inyecciones , Inyecciones Intraperitoneales , Factor I del Crecimiento Similar a la Insulina/genética , Coagulación con Láser , Pulmón/citología , Ratones , Ratones Endogámicos C57BL , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/genética , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , ARN Mensajero/metabolismo , Receptores CXCR4/antagonistas & inhibidores , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor A de Crecimiento Endotelial Vascular/genética , Cuerpo VítreoRESUMEN
BACKGROUND: Treatment of alopecia areata with intralesional steroid injection is generally recommended for people who have less than 50% scalp involvement. In a specialized hair loss clinic, the authors successfully treated patients with extensive alopecia areata (over 50% but under 99%) with intralesional corticosteroid injections. OBSERVATIONS: A review of patients with extensive alopecia areata was done. Six out of 10 patients responded to treatment with intralesional triamcinolone acetonide. In comparison to the non-responders, the responders tended to have exclamation mark hairs and a positive hair pull test at their initial physical examination, and exhibited improvement during the initial months of treatment. Complications were negligible, with mild reversible atrophy in three patients. The treatment was well tolerated and, in some patients, pain was minimized by use of a topical anesthetic agent applied under occlusion prior to the visit. CONCLUSION: Intralesional triamcinolone acetonide is a safe and effective treatment for patients with extensive alopecia areata. Patients with exclamation point hairs and a positive hair pull test may be more likely to respond.
Asunto(s)
Alopecia Areata/tratamiento farmacológico , Glucocorticoides/uso terapéutico , Triamcinolona Acetonida/uso terapéutico , Adolescente , Adulto , Anestésicos Locales/uso terapéutico , Femenino , Glucocorticoides/administración & dosificación , Glucocorticoides/efectos adversos , Humanos , Inyecciones Intralesiones , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Índice de Severidad de la Enfermedad , Resultado del Tratamiento , Triamcinolona Acetonida/administración & dosificación , Triamcinolona Acetonida/efectos adversosRESUMEN
RATIONALE: Insulin-like growth factor binding protein (IGFBP)-3 modulates vascular development by regulating endothelial progenitor cell (EPC) behavior, specifically stimulating EPC cell migration. This study was undertaken to investigate the mechanism of IGFBP-3 effects on EPC function and how IGFBP-3 mediates cytoprotection following vascular injury. OBJECTIVE: To examine the mechanism of IGFBP-3-mediated repair following vascular injury. METHODS AND RESULTS: We used 2 complementary vascular injury models: laser occlusion of retinal vessels in adult green fluorescent protein (GFP) chimeric mice and oxygen-induced retinopathy in mouse pups. Intravitreal injection of IGFBP-3-expressing plasmid into lasered GFP chimeric mice stimulated homing of EPCs, whereas reversing ischemia induced increases in macrophage infiltration. IGFBP-3 also reduced the retinal ceramide/sphingomyelin ratio that was increased following laser injury. In the OIR model, IGFBP-3 prevented cell death of resident vascular endothelial cells and EPCs, while simultaneously increasing astrocytic ensheathment of vessels. For EPCs to orchestrate repair, these cells must migrate into ischemic tissue. This migratory ability is mediated, in part, by endogenous NO generation. Thus, we asked whether the migratory effects of IGFBP-3 were attributable to stimulation of NO generation. IGFBP-3 increased endothelial NO synthase expression in human EPCs leading to NO generation. IGFBP-3 exposure also led to the redistribution of vasodilator-stimulated phosphoprotein, an NO regulated protein critical for cell migration. IGFBP-3-mediated NO generation required high-density lipoprotein receptor activation and stimulation of phosphatidylinositol 3-kinase/Akt pathway. CONCLUSION: These studies support consideration of IGFBP-3 as a novel agent to restore the function of injured vasculature and restore NO generation.
Asunto(s)
Movimiento Celular , Células Endoteliales/metabolismo , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/metabolismo , Óxido Nítrico/metabolismo , Neovascularización Retiniana/metabolismo , Vasos Retinianos/metabolismo , Retinopatía de la Prematuridad/metabolismo , Células Madre/metabolismo , Animales , Animales Recién Nacidos , Astrocitos/metabolismo , Moléculas de Adhesión Celular/metabolismo , Muerte Celular , Proliferación Celular , Células Cultivadas , Ceramidas/metabolismo , Arterias Cerebrales/metabolismo , Arterias Cerebrales/fisiopatología , Citoprotección , Modelos Animales de Enfermedad , Células Endoteliales/patología , Femenino , Técnicas de Transferencia de Gen , Proteínas Fluorescentes Verdes/genética , Humanos , Recién Nacido , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/genética , Masculino , Arterias Mesentéricas/metabolismo , Arterias Mesentéricas/fisiopatología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Proteínas de Microfilamentos/metabolismo , Óxido Nítrico Sintasa de Tipo III/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Fosfoproteínas/metabolismo , Embarazo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ratas , Ratas Sprague-Dawley , Neovascularización Retiniana/patología , Neovascularización Retiniana/fisiopatología , Vasos Retinianos/patología , Vasos Retinianos/fisiopatología , Retinopatía de la Prematuridad/patología , Retinopatía de la Prematuridad/fisiopatología , Receptores Depuradores de Clase B/metabolismo , Transducción de Señal , Esfingomielinas/metabolismo , Células Madre/patología , VasodilataciónRESUMEN
Precise localization of exogenously delivered stem cells is critical to our understanding of their reparative response. Our current inability to determine the exact location of small numbers of cells may hinder optimal development of these cells for clinical use. We describe a method using magnetic resonance imaging to track and localize small numbers of stem cells following transplantation. Endothelial progenitor cells (EPC) were labeled with monocrystalline iron oxide nanoparticles (MIONs) which neither adversely altered their viability nor their ability to migrate in vitro and allowed successful detection of limited numbers of these cells in muscle. MION-labeled stem cells were also injected into the vitreous cavity of mice undergoing the model of choroidal neovascularization, laser rupture of Bruch's membrane. Migration of the MION-labeled cells from the injection site towards the laser burns was visualized by MRI. In conclusion, MION labeling of EPC provides a non-invasive means to define the location of small numbers of these cells. Localization of these cells following injection is critical to their optimization for therapy.
Asunto(s)
Medios de Contraste/metabolismo , Imagen por Resonancia Magnética/métodos , Coloración y Etiquetado/métodos , Células Madre/metabolismo , Apoptosis/efectos de los fármacos , Adhesión Celular/fisiología , Diferenciación Celular/fisiología , Movimiento Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Materiales Biocompatibles Revestidos/metabolismo , Colorantes/metabolismo , Relación Dosis-Respuesta a Droga , Ferrocianuros/metabolismo , Óxido Ferrosoférrico/metabolismo , Óxido Ferrosoférrico/farmacología , Fibronectinas/metabolismo , Humanos , Nanopartículas , Células Madre/citología , Células Madre/fisiologíaRESUMEN
Rapid and accurate detection of pathogenic bacteria is important for the treatment of patients with suitable antibiotics. Here we report the development of a diagnostic DNA microarray for the high-throughput identification of 39 pathogenic bacteria selected based on their high prevalence rate and/or difficulty of cultivation. The 23S ribosomal DNA and 16S-23S rDNA intergenic spacer region were used as target DNAs for pathogen detection. Universal- and species-specific probes were designed based on the unique and common sites within the target DNA sequences. New target DNA sequences were determined for the detection of 19 bacterial pathogens. The usefulness of the designed probes was validated using 39 reference bacteria and also with 515 clinical isolates from various clinical samples including blood, stool, pus, sputum, urine and cerebrospinal fluid. The DNA microarray developed in this study allowed efficient detection of bacterial pathogens with the specificities of 100%. The sensitivities were 100% as well except for the two pathogens, Enterobacter cloacae (75%) and Enterococcus faecium (85%). These results suggest that the DNA microarray-based assay developed in this study outperforms current diagnostic systems with respect to sensitivity, specificity, and high-throughput detection, and thus should be useful in pathogen diagnosis in the clinical setting.
Asunto(s)
Bacterias/genética , Bacterias/aislamiento & purificación , Infecciones Bacterianas/diagnóstico , Infecciones Bacterianas/microbiología , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Bacterias/patogenicidad , Enfermedades Transmisibles/microbiología , ADN Espaciador Ribosómico/genética , Enterobacter cloacae/genética , Enterobacter cloacae/aislamiento & purificación , Enterobacter cloacae/patogenicidad , Enterococcus faecium/genética , Enterococcus faecium/aislamiento & purificación , Enterococcus faecium/patogenicidad , Humanos , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: We examined the effect of the vasoactive agents carbon monoxide (CO) and nitric oxide (NO) : n the phosphorylation and intracellular redistribution of vasodilator-stimulated phosphoprotein (VASP), a critical actin motor protein required for cell migration that also controls vasodilation and platelet aggregation. RESEARCH DESIGN AND METHODS: We examined the effect of donor-released CO and NO in endothelial progenitor cells (EPCs) and platelets from nondiabetic and diabetic subjects and in human microvascular endothelial cells (HMECs) cultured under low (5.5 mmol/l) or high (25 mmol/l) glucose conditions. VASP phosphorylation was evaluated using phosphorylation site-specific antibodies. RESULTS: In control platelets, CO selectively promotes phosphorylation at VASP Ser-157, whereas NO promotes phosphorylation primarily at Ser-157 and also at Ser-239, with maximal responses at 1 min with both agents on Ser-157 and at 15 min on Ser-239 with NO treatment. In diabetic platelets, neither agent resulted in VASP phosphorylation. In nondiabetic EPCs, NO and CO increased phosphorylation at Ser-239 and Ser-157, respectively, but this response was markedly reduced in diabetic EPCs. In endothelial cells cultured under low glucose conditions, both CO and NO induced phosphorylation at Ser-157 and Ser-239; however, this response was completely lost when cells were cultured under high glucose conditions. In control EPCs and in HMECs exposed to low glucose, VASP was redistributed to filopodia-like structures following CO or NO exposure; however, redistribution was dramatically attenuated under high glucose conditions. CONCLUSIONS: Vasoactive gases CO and NO promote cytoskeletal changes through site- and cell type-specific VASP phosphorylation, and in diabetes, blunted responses to these agents may lead to reduced vascular repair and tissue perfusion.