RESUMEN

Clinical management of delayed healing or non-union of long bone fractures and segmental defects poses a substantial orthopaedic challenge. There are suggestions in the literature that bone healing may be enhanced by inhibiting the activities of T and B lymphocytes, but this remains controversial. To examine this matter in more detail, sub-critical-sized segmental defects were created in the femora of mice and it was assessed whether there might be a benefit from the administration of a Food and Drug Administration (FDA)-approved drug that blocks T cell activation (tacrolimus). Defects were stabilised using an internal plate. In certain groups of animals, 1 mg/kg or 10 mg/kg tacrolimus was delivered locally to the defect site for 3 or 7 d using an implanted osmotic pump with a silicon catheter directing drug delivery into the defect area. Healing was monitored by weekly X-ray and assessed at 12 weeks by mechanical testing, µCT and histology. Radiographic and histological evaluations revealed that 100 % of defects healed well regardless of tacrolimus dosage or duration. A comparison of healed C57BL/6 and Rag1-/- femora by µCT and ex vivo torsion testing showed no differences within mouse strains in terms of bone volume, tissue volume, bone volume/tissue volume ratio, shear modulus, torsional rigidity or torsional stiffness. These data failed to support an important role for tacrolimus in modulating the natural healing of segmental defects under those experimental conditions.

Asunto(s)

Curación de Fractura/efectos de los fármacos , Fracturas Óseas/tratamiento farmacológico , Fracturas Óseas/metabolismo , Proteínas de Homeodominio/metabolismo , Tacrolimus/farmacología , Animales , Linfocitos B/efectos de los fármacos , Linfocitos B/metabolismo , Fémur , Masculino , Ratones , Ratones Endogámicos C57BL , Osteotomía/métodos , Linfocitos T/efectos de los fármacos , Linfocitos T/metabolismo , Microtomografía por Rayos X/métodos

RESUMEN

Mobilized blood has supplanted bone marrow (BM) as the primary source of hematopoietic stem cells for autologous and allogeneic stem cell transplantation. Pharmacologically enforced egress of hematopoietic stem cells from BM, or mobilization, has been achieved by directly or indirectly targeting the CXCL12/CXCR4 axis. Shortcomings of the standard mobilizing agent, granulocyte colony-stimulating factor (G-CSF), administered alone or in combination with the only approved CXCR4 antagonist, Plerixafor, continue to fuel the quest for new mobilizing agents. Using Protein Epitope Mimetics technology, a novel peptidic CXCR4 antagonist, POL5551, was developed. In vitro data presented herein indicate high affinity to and specificity for CXCR4. POL5551 exhibited rapid mobilization kinetics and unprecedented efficiency in C57BL/6 mice, exceeding that of Plerixafor and at higher doses also of G-CSF. POL5551-mobilized stem cells demonstrated adequate transplantation properties. In contrast to G-CSF, POL5551 did not induce major morphological changes in the BM of mice. Moreover, we provide evidence of direct POL5551 binding to hematopoietic stem and progenitor cells (HSPCs) in vivo, strengthening the hypothesis that CXCR4 antagonists mediate mobilization by direct targeting of HSPCs. In summary, POL5551 is a potent mobilizing agent for HSPCs in mice with promising therapeutic potential if these data can be corroborated in humans.

Asunto(s)

Células Madre Hematopoyéticas/efectos de los fármacos , Proteínas/farmacología , Receptores CXCR4/antagonistas & inhibidores , Animales , Microambiente Celular , Sinergismo Farmacológico , Factor Estimulante de Colonias de Granulocitos/farmacología , Células Madre Hematopoyéticas/citología , Macrófagos/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Osteoblastos/efectos de los fármacos

RESUMEN

Receptor activator of nuclear factor-kappaB-ligand (RANKL), encoded by the gene TNFSF11, is required for osteoclastogenesis, and its expression is upregulated in pathologic bone loss. Transcript variants of TNFSF11 messenger RNA (mRNA) have been described that encode a membrane-bound and a putative secreted form of RANKL. We identify a TNFSF11 transcript variant that extends the originally identified transcript encoding secreted RANKL. We demonstrate that this TNFSF11 transcript variant is expressed by the human osteosarcoma cell line, Saos-2, and by both primary human T cells and Jurkat T cells. Of relevance to the production of RANKL in pathologic bone loss, expression of this secreted TNFSF11 transcript is upregulated in Jurkat T cells and primary human T cells upon activation. Furthermore, this transcript can be translated and secreted in Jurkat T cells in vitro and is able to support osteoclast differentiation. Our data highlight the complexity of the TNFSF11 genomic locus, and demonstrate the potential for the expression of alternate mRNA transcripts encoding membrane-bound and secreted forms of RANKL. Implications of alternate mRNA transcripts encoding different RANKL protein isoforms should be carefully considered and specifically examined in future studies, particularly those implicating RANKL in pathologic bone loss.

Asunto(s)

Empalme Alternativo , Ligando RANK/genética , ARN Mensajero/genética , Linfocitos T/metabolismo , Animales , Western Blotting , Diferenciación Celular/efectos de los fármacos , Línea Celular Tumoral , Células Cultivadas , Humanos , Células Jurkat , Activación de Linfocitos , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Osteoclastos/citología , Osteoclastos/efectos de los fármacos , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Isoformas de Proteínas/farmacología , Ligando RANK/metabolismo , Ligando RANK/farmacología , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Regulación hacia Arriba

RESUMEN

SETTING: A key program performance objective established by the Centers for Disease Control and Prevention (CDC) is that ≥93% of tuberculosis (TB) cases complete treatment within 12 months. OBJECTIVE: To determine the rate of and risk factors for delay in anti-tuberculosis treatment completion. DESIGN: Nested case-control study among TB cases reported to the Tennessee Department of Health between 1 January 2000 and 31 December 2010. Time to complete treatment was calculated using treatment start and stop dates documented in the Tuberculosis Information Management System (TIMS). RESULTS: Of 2627 cases, 261 (9.9%) required >12 months to complete treatment. In adjusted conditional logistic regression analyses, cavitary disease and positive cultures after 2 months of therapy (OR 5.85, 95%CI 1.98-17.32, P = 0.001), non-adherence (OR 4.13, 95%CI 1.76-9.72, P < 0.001), and interruptions in treatment due to drug-related issues (OR 6.91, 95%CI 3.76-12.70, P < 0.001) were independently associated with delay in completion of TB treatment. CONCLUSION: From 2000 to 2010, the proportion of TB cases completing treatment within 12 months increased from 84.6% to 94.9%, and remained above the CDC target during 2009-2010. Efforts to improve patient adherence and reduce interruptions in treatment due to anti-tuberculosis drug-related issues could improve the proportion of TB cases completing treatment within 12 months.

Asunto(s)

Antituberculosos/efectos adversos , Cumplimiento de la Medicación , Tuberculosis/tratamiento farmacológico , Adulto , Estudios de Casos y Controles , Terapia por Observación Directa , Femenino , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Evaluación de Programas y Proyectos de Salud , Factores de Riesgo , Tennessee , Factores de Tiempo

RESUMEN

The CXCR4 antagonist AMD3100 is progressively replacing cyclophosphamide (CYP) as adjuvant to granulocyte colony-stimulating factor (G-CSF) to mobilize hematopoietic stem cells (HSC) for autologous transplants in patients who failed prior mobilization with G-CSF alone. It has recently emerged that G-CSF mediates HSC mobilization and inhibits bone formation via specific bone marrow (BM) macrophages. We compared the effect of these three mobilizing agents on BM macrophages, bone formation, osteoblasts, HSC niches and HSC reconstitution potential. Both G-CSF and CYP suppressed niche-supportive macrophages and osteoblasts, and inhibited expression of endosteal cytokines resulting in major impairment of HSC reconstitution potential remaining in the mobilized BM. In sharp contrast, although AMD3100 was effective at mobilizing HSC, it did not suppress osteoblasts, endosteal cytokine expression or reconstitution potential of HSC remaining in the mobilized BM. In conclusion, although G-CSF, CYP and AMD3100 efficiently mobilize HSC into the blood, their effects on HSC niches and bone formation are distinct with both G-CSF and CYP targeting HSC niche function and bone formation, whereas AMD3100 directly targets HSC without altering niche function or bone formation.

Asunto(s)

Médula Ósea/efectos de los fármacos , Ciclofosfamida/farmacología , Factor Estimulante de Colonias de Granulocitos/farmacología , Hematínicos/farmacología , Células Madre Hematopoyéticas/efectos de los fármacos , Compuestos Heterocíclicos/farmacología , Osteogénesis/efectos de los fármacos , Animales , Fármacos Anti-VIH/farmacología , Antineoplásicos Alquilantes/farmacología , Bencilaminas , Médula Ósea/metabolismo , Células Cultivadas , Ciclamas , Citometría de Flujo , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/metabolismo , Técnicas para Inmunoenzimas , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Osteoblastos/citología , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa

RESUMEN

Although opiate drugs of abuse have been shown to decrease adult hippocampal neurogenesis, the impact of opiate analgesics has not been tested. North American regulatory boards governing the ethical treatment of experimental animals require the administration of analgesics, such as buprenorphine, following minor surgical interventions. Here, we show that two commonly used post-operative buprenorphine dosing regimes significantly inhibit the proliferation of doublecortin-positive neuroblasts but not other hippocampal stem and progenitor cell populations in adult mice. Buprenorphine, administered in schedules of three 0.05 mg/kg subcutaneous injections over a single day or seven 0.05 mg/kg injections over a 3-day period decreased the number of actively proliferating 5-iodo-2'-deoxyuridine-labeled doublecortin-positive cells for up to 6 days after opiate withdrawal. The minimal (three injection), but not standard (seven injection), analgesic paradigm also reduced basal indices of hippocampal progenitor cell apoptosis and enhanced survival of newly born cells for up to 28 days. Taken together, these data provide the first evidence that the routine administration of opiate analgesics has transient but long-lasting effects on neurogenesis and further emphasize that analgesic dosage and schedule should be reported and considered when interpreting the magnitude of neural stem and progenitor cell activation in response to in vivo intervention.

Asunto(s)

Células Madre Adultas/efectos de los fármacos , Analgésicos/farmacología , Buprenorfina/farmacología , Proliferación Celular/efectos de los fármacos , Hipocampo/citología , Neurogénesis/efectos de los fármacos , Análisis de Varianza , Animales , Antígenos/metabolismo , Bromodesoxiuridina/metabolismo , Supervivencia Celular/efectos de los fármacos , Desoxiuridina/metabolismo , Proteínas de Dominio Doblecortina , Proteína Ácida Fibrilar de la Glía , Hipocampo/efectos de los fármacos , Etiquetado Corte-Fin in Situ , Masculino , Ratones , Ratones Endogámicos C57BL , Proteínas Asociadas a Microtúbulos/metabolismo , Neuroglía/metabolismo , Neuropéptidos/metabolismo , Fosfopiruvato Hidratasa/metabolismo , Proteoglicanos/metabolismo , Factores de Tiempo

RESUMEN

SETTING: Programmatic data from the United States on tuberculosis (TB) recurrence are limited. OBJECTIVES: To determine the TB recurrence rate and to determine if chronic lung disease (CLD) and human immunodeficiency virus (HIV) infection are risk factors for recurrence in this population. DESIGN: Nested case-control study among TB cases reported to the Tennessee Department of Health between 1 January 2000 and 31 December 2006. Time at risk for recurrence was through 31 December 2007. Multiple imputation accounted for missing data. RESULTS: Of 1431 TB cases, 20 cases recurred (1.4%, 95%CI 0.9-2.1). Median time at risk for recurrence was 4.5 years (interquartile range 2.7-6.1). Initial and recurrent Mycobacterium tuberculosis isolates were available for genotyping for 15 patients; 12 were consistent with relapse (0.8%, 95%CI 0.4-1.5) and three with re-infection (0.2%, 95%CI 0.04-0.6). HIV infection (OR 5.01, P = 0.04) and CLD (OR 5.28, P = 0.03) were independently associated with recurrent TB, after adjusting for a disease risk score. HIV infection was a risk factor for TB re-infection (P < 0.001). CONCLUSIONS: In this low-incidence US population, the TB recurrence rate was low, but CLD and HIV were independent risk factors for recurrence. HIV infection was also a risk factor for TB re-infection.

Asunto(s)

Infecciones por VIH/complicaciones , Enfermedades Pulmonares/complicaciones , Tuberculosis/epidemiología , Adulto , Estudios de Casos y Controles , Enfermedad Crónica , Femenino , Infecciones por VIH/epidemiología , Humanos , Enfermedades Pulmonares/epidemiología , Masculino , Persona de Mediana Edad , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/aislamiento & purificación , Recurrencia , Factores de Riesgo , Tennessee/epidemiología , Factores de Tiempo , Tuberculosis/etiología , Tuberculosis/microbiología

RESUMEN

OBJECTIVE: To characterize the proportion of tuberculosis (TB) cases that could have been prevented among human immunodeficiency virus (HIV) infected persons receiving care in the era of highly active antiretroviral treatment (HAART). DESIGN: We conducted an observational cohort study among HIV-infected patients with >or=2 out-patient visits at the Comprehensive Care Center, Nashville, Tennessee, USA, between 1 January 1998 and 31 December 2005. METHODS: A potentially preventable TB case was defined as a case in which the patient received no screening tuberculin skin test (TST) prior to TB diagnosis or a case in which a patient with a positive screening TST did not complete treatment for latent infection. RESULTS: Of 3601 HIV-infected persons in care (13 905 person-years [p-y] of follow-up), 29 developed TB (230/100,000 p-y). Of the 29, 20 (69%) had not had TST performed as part of routine screening. Of the nine patients screened, four had a positive test, three of whom completed treatment for latent TB infection. Of 29 TB cases, 21 (72%) were therefore potentially preventable. CONCLUSIONS: Most TB cases in this cohort were potentially preventable had the patients undergone a screening TST followed by treatment of latent infection if they had a positive TST.

Asunto(s)

Infecciones por VIH/epidemiología , Tuberculosis Pulmonar/prevención & control , Adulto , Terapia Antirretroviral Altamente Activa , Comorbilidad , Femenino , Infecciones por VIH/tratamiento farmacológico , Humanos , Masculino , Persona de Mediana Edad , Prueba de Tuberculina

RESUMEN

Effective bone biomaterials provide structural support for bone regeneration and elicit minimal inflammatory or toxic effects in vivo. Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a bacterially derived biodegradable polymer that possesses suitable mechanical strength for use as a bone biomaterial and has a slow rate of degradation in biological environments. Our previous in vitro study showed that many PHBV preparations are contaminated with bacterial lipopolysaccharide, and we developed a purification procedure to substantially remove it. Here, we have evaluated the in vivo biocompatibility of PHBV purified by H(2)O(2) treatment and solvent extraction. We utilized a murine tibial defect model consisting of a hole drilled through the diameter of the tibial diaphysis into which nonporous cylindrical plugs of purified PHBV were implanted. The animals were sacrificed at 1 week and 4 weeks postsurgery, and tibiae were examined using histological staining. The PHBV implant induced a mild inflammatory response 1 week after injury, which persisted for 4 weeks. Granuloma type tissues formed only when the implant protruded into the overlaying tissue. Woven bone formation occurred adjacent to the implant, which gave rise to lamellar bone and stabilized the implant indicating that the PHBV did not affect this process. Our data validated the murine defect model and indicate that solid PHBV induces a mild tissue reaction with bone deposition adjacent to the implant with no fibrous tissue present at 4 weeks post surgery.

Asunto(s)

Materiales Biocompatibles/química , Regeneración Ósea , Sustitutos de Huesos , Poliésteres/química , Prótesis e Implantes , Tibia/metabolismo , Animales , Bacterias/metabolismo , Huesos/metabolismo , Peróxido de Hidrógeno/química , Lipopolisacáridos/metabolismo , Masculino , Ratones , Microscopía Electrónica de Rastreo/métodos , Membrana Mucosa/metabolismo , Solventes/química

RESUMEN

Young Australian Aboriginal children in remote communities experience very high rates of pneumococcal carriage and otitis media. Prior to introduction of the 7-valent pneumococcal conjugate vaccine (7vPCV, Prevenar), serotype 16F was an important type found in nasal and ear discharge swabs. Since commencement of pneumococcal immunisation for Aboriginal infants in 2001, 16F has become the predominant established serotype in carriage and otitis media in young Aboriginal children. BOX typing and multi-locus sequence typing revealed a diverse population of serotype 16F strains, and evidence of potential capsule switching from a vaccine serotype 4 to a serotype 16F.

Asunto(s)

Vacunas Meningococicas/uso terapéutico , Otitis Media/microbiología , Otitis Media/prevención & control , Infecciones Neumocócicas/microbiología , Infecciones Neumocócicas/prevención & control , Vacunas Neumococicas/uso terapéutico , Streptococcus pneumoniae/clasificación , Australia , Cápsulas Bacterianas , Niño , Vacuna Neumocócica Conjugada Heptavalente , Humanos , Nativos de Hawái y Otras Islas del Pacífico , Serotipificación , Streptococcus pneumoniae/aislamiento & purificación

RESUMEN

OBJECTIVES: Receptor activator of NF-kappaB ligand (RANKL) and osteoprotegerin (OPG) have been demonstrated to be critical regulators of osteoclast generation and activity. In addition, RANKL has been implicated as an important mediator of bone erosion in rheumatoid arthritis (RA). However, the expression of RANKL and OPG at sites of pannus invasion into bone has not been examined. The present study was undertaken to further elucidate the contribution of this cytokine system to osteoclastogenesis and subsequent bone erosion in RA by examining the pattern of protein expression for RANKL, OPG and the receptor activator of NF-kappaB (RANK) in RA at sites of articular bone erosion. METHODS: Tissues from 20 surgical procedures from 17 patients with RA were collected as discarded materials. Six samples contained only synovium or tenosynovium remote from bone, four samples contained pannus-bone interface with adjacent synovium and 10 samples contained both synovium remote from bone and pannus-bone interface with adjacent synovium. Immunohistochemistry was used to characterize the cellular pattern of RANKL, RANK and OPG protein expression immediately adjacent to and remote from sites of bone erosion. RESULTS: Cellular expression of RANKL protein was relatively restricted in the bone microenvironment; staining was focal and confined largely to sites of osteoclast-mediated erosion at the pannus-bone interface and at sites of subchondral bone erosion. RANK-expressing osteoclast precursor cells were also present in these sites. OPG protein expression was observed in numerous cells in synovium remote from bone but was more limited at sites of bone erosion, especially in regions associated with RANKL expression. CONCLUSIONS: The pattern of RANKL and OPG expression and the presence of RANK-expressing osteoclast precursor cells at sites of bone erosion in RA contributes to the generation of a local microenvironment that favours osteoclast differentiation and activity. These data provide further evidence implicating RANKL in the pathogenesis of arthritis-induced joint destruction.

Asunto(s)

Artritis Reumatoide/metabolismo , Resorción Ósea/metabolismo , Proteínas Portadoras/análisis , Articulaciones/química , Glicoproteínas de Membrana/análisis , Membrana Sinovial/química , Adolescente , Adulto , Artritis Reumatoide/patología , Resorción Ósea/patología , Diferenciación Celular , Niño , Preescolar , Glicoproteínas/análisis , Humanos , Inmunohistoquímica/métodos , Articulaciones/patología , Osteoclastos/patología , Osteoprotegerina , Ligando RANK , Receptor Activador del Factor Nuclear kappa-B , Receptores Citoplasmáticos y Nucleares/análisis , Receptores del Factor de Necrosis Tumoral/análisis , Membrana Sinovial/metabolismo

RESUMEN

The aim of this study was to compare transcriptional regulation in vivo during anabolic bone formation induced by either estradiol (E2) treatment or intermittent parathyroid hormone[1-34] (PTH) therapy. We utilized an ovariectomized (OVX) mouse model of osteoporosis and transcriptional profiling to identify genes upregulated by either high-dose E2 or PTH. Five weeks post-OVX, the mice were administered either E2 and/or PTH, or vehicle for 4 weeks. Femoral bones were analyzed by microCT and histomorphometry to confirm the anabolic effect of each treatment. OVX vehicle-treated control mice lost metaphyseal trabecular bone, with significant decrease in trabecular number, thickness, and connectivity. Both E2 and PTH treatments increased trabecular and cortical bone indices above the level of the sham operated controls, fully restoring both bone volume and bone mineral density (BMD). Moreover, PTH/E2 combination treatment led to significantly greater increase in cancellous bone and BMD than would be expected from the additive effects of the separate treatments. To determine whether PTH and E2 treatments were stimulating similar bone anabolic mechanisms, or were activating distinct signaling pathways, we compared patterns of gene expression using transcriptional profiling after either E2 or PTH treatment. After 4, 11, and 24 days of treatment, total RNA was collected from both the distal femoral metaphysis and diaphysis. Transcriptional profiling was performed using Affymetrix GeneChip probe arrays, comprised of approximately 36,000 full-length mouse genes and EST clusters from the UniGene database. Several markers of osteoblast activity, including c-fos, RANKL, PHEX, and PTHR1, were consistently upregulated by PTH in both skeletal sites. PTH treatment also increased expression of Cathespin K, consistent with the predicted increase in osteoclast activity. E2 treatment upregulated a largely distinct set of genes, including TGFbeta3, and BMP1, as well as several genes critical for cell cycle control, including Cyclin D1 and CDK inhibitor 1A. Overall, comparison of transcriptional profiles suggest that anabolic responses in bone to PTH and high-dose E2 treatment after OVX-induced osteoporosis involve largely distinct patterns of gene regulation, each resulting in restoration of bone mass.

Asunto(s)

Densidad Ósea/fisiología , Resorción Ósea/metabolismo , Huesos/metabolismo , Estradiol/farmacología , Hormona Paratiroidea/farmacología , Absorciometría de Fotón , Animales , Densidad Ósea/efectos de los fármacos , Proteína Morfogenética Ósea 1 , Proteínas Morfogenéticas Óseas/metabolismo , Resorción Ósea/diagnóstico por imagen , Resorción Ósea/tratamiento farmacológico , Huesos/diagnóstico por imagen , Huesos/efectos de los fármacos , Catepsina K , Catepsinas/metabolismo , Femenino , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , Genes cdc/fisiología , Metaloendopeptidasas/metabolismo , Ratones , Osteoporosis/diagnóstico por imagen , Osteoporosis/tratamiento farmacológico , Osteoporosis/metabolismo , Ovariectomía/métodos , Factor de Crecimiento Transformador beta/metabolismo , Factor de Crecimiento Transformador beta3

RESUMEN

OBJECTIVES: To examine the potential role of the angiogenic growth factor angiopoietin-1 (Ang-1) in inflammatory arthritis. METHODS: Eighteen synovial tissue samples were obtained from 17 patients with a clinical diagnosis of rheumatoid arthritis (RA) and compared with six synovial tissue samples from six patients with osteoarthritis (OA). Ang-1 expression in synovial tissues was determined by immunohistochemistry and in situ hybridisation. Ang-1 mRNA and protein expression were also examined by northern blot analysis and enzyme linked immunosorbent assay (ELISA) in cultured synovial fibroblasts and human umbilical vein endothelial cells (HUVECs) before and after treatment with tumour necrosis factor (TNF)alpha. RESULTS: Ang-1 protein expression was detected by immunohistochemistry in 16/18 RA synovial tissue samples. Ang-1 protein was frequently observed in the synovial lining layer and in cells within the sublining synovial tissue, in both perivascular areas and in areas remote from vessels. In contrast, Ang-1 was only weakly detected in these sites in OA samples. Ang-1 mRNA and protein were also expressed in cultured synovial fibroblasts derived from patients with RA. In addition, induction of Ang-1 mRNA and protein was observed by northern blot analysis and ELISA after stimulation of RA synovial fibroblasts, but not HUVECs, with the proinflammatory cytokine TNF alpha. CONCLUSIONS: Ang-1 mRNA and protein are expressed in the synovium of patients with RA, and are up regulated in synovial fibroblasts by TNF alpha. Ang-1 may therefore be an important regulator of angiogenesis in inflammatory arthritis.

Asunto(s)

Inductores de la Angiogénesis/metabolismo , Artritis Reumatoide/metabolismo , Glicoproteínas de Membrana/metabolismo , Membrana Sinovial/metabolismo , Inductores de la Angiogénesis/genética , Angiopoyetina 1 , Angiopoyetina 2 , Artritis Reumatoide/patología , Northern Blotting/métodos , Técnicas de Cultivo , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Humanos , Técnicas para Inmunoenzimas , Hibridación in Situ , Interleucina-1/farmacología , Glicoproteínas de Membrana/genética , Osteoartritis/metabolismo , ARN Mensajero/genética , Membrana Sinovial/irrigación sanguínea , Factor de Necrosis Tumoral alfa/farmacología

RESUMEN

Depression is a serious, often chronic disease that can be managed effectively with a chronic care model in primary care settings. Depressed persons are likely to be seen by a primary care physician, but their condition often goes unrecognized and untreated. There are effective treatment models that consist of efficacious psychotherapeutic and pharmacological interventions, use of evidence-based guidelines for primary care treatment of depression, development of explicit plans and protocols, reorganization of practice, longitudinal follow-up, patient self-management, decision-making support, access to community resources and leadership commitment. Moving these models into everyday practice requires overcoming both clinical and system barriers. Barriers consist of issues surrounding patients, providers, practices, plans, and purchasers. An understanding of these barriers at each level helps to provide a framework for the changes required to overcome them. The Robert Wood Johnson Foundation National Program on Depression in Primary Care will seek to apply simultaneously both clinical and system strategies in a new five-year initiative to overcome these barriers.

Asunto(s)

Antidepresivos/uso terapéutico , Continuidad de la Atención al Paciente/organización & administración , Trastorno Depresivo/terapia , Accesibilidad a los Servicios de Salud/organización & administración , Servicios de Salud Mental/organización & administración , Atención Primaria de Salud/normas , Enfermedad Crónica , Toma de Decisiones , Trastorno Depresivo/tratamiento farmacológico , Humanos , Relaciones Médico-Paciente , Psicoterapia , Estados Unidos

RESUMEN

There is considerable evidence that osteoclasts are involved in the pathogenesis of focal bone erosion in rheumatoid arthritis. Tumor necrosis factor-related activation-induced cytokine, also known as receptor activator of nuclear factor-kappaB ligand (TRANCE/RANKL) is an essential factor for osteoclast differentiation. In addition to its role in osteoclast differentiation and activation, TRANCE/RANKL also functions to augment T-cell dendritic cell cooperative interactions. To further evaluate the role of osteoclasts in focal bone erosion in arthritis, we generated inflammatory arthritis in the TRANCE/RANKL knockout mouse using a serum transfer model that bypasses the requirement for T-cell activation. These animals exhibit an osteopetrotic phenotype characterized by the absence of osteoclasts. Inflammation, measured by clinical signs of arthritis and histopathological scoring, was comparable in wild-type and TRANCE/RANKL knockout mice. Microcomputed tomography and histopathological analysis demonstrated that the degree of bone erosion in TRANCE/RANKL knockout mice was dramatically reduced compared to that seen in control littermate mice. In contrast, cartilage erosion was present in both control littermate and TRANCE/RANKL knockout mice. These results confirm the central role of osteoclasts in the pathogenesis of bone erosion in arthritis and demonstrate distinct mechanisms of cartilage destruction and bone erosion in this animal model of arthritis.

Asunto(s)

Artritis/diagnóstico , Artritis/metabolismo , Huesos/diagnóstico por imagen , Huesos/patología , Glicoproteínas de Membrana/deficiencia , Animales , Artritis/sangre , Artritis/etiología , Transfusión Sanguínea , Proteínas Portadoras/genética , Proteínas Portadoras/fisiología , Cartílago Articular/diagnóstico por imagen , Cartílago Articular/patología , Modelos Animales de Enfermedad , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/fisiología , Ratones , Ratones Noqueados/genética , Ligando RANK , Receptor Activador del Factor Nuclear kappa-B , Tomografía Computarizada por Rayos X

RESUMEN

OBJECTIVES: To compare immunohistochemical scoring with clinical scoring and radiology for the assessment of rheumatoid arthritis (RA) disease activity, synovial tissue (ST) biopsied arthroscopically was assessed from 18 patients before and after commencement of disease-modifying anti-rheumatic drug (DMARD) therapy. METHODS: Lymphocytes, macrophages, differentiated dendritic cells (DC), vascularity, tumour necrosis factor (TNF) alpha and interleukin-1beta levels were scored. Clinical status was scored using the American College of Rheumatology (ACR) core set and serial radiographs were scored using the Larsen and Sharp methods. Histopathological evidence of activity included infiltration by lymphocytes, DC, macrophages, tissue vascularity, and expression of lining and sublining TNFalpha. These indices co-varied across the set of ST biopsies and were combined as a synovial activity score for each biopsy. RESULTS: The change in synovial activity with treatment correlated with the ACR clinical response and with decreased radiological progression by the Larsen score. The ACR response to DMARD therapy, the change in synovial activity score and the slowing of radiological progression were each greatest in patients with high initial synovial vascularity. CONCLUSIONS: The data demonstrate an association between clinical, radiological and synovial immunopathological responses to anti-rheumatic treatment in RA. High ST vascularity may predict favourable clinical and radiological responses to treatment.

Asunto(s)

Antirreumáticos/uso terapéutico , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/patología , Membrana Sinovial/patología , Anciano , Artritis Reumatoide/diagnóstico por imagen , Artritis Reumatoide/inmunología , Biopsia , Células Dendríticas/inmunología , Humanos , Interleucina-1/análisis , Linfocitos/inmunología , Macrófagos/inmunología , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Radiografía , Membrana Sinovial/irrigación sanguínea , Membrana Sinovial/inmunología , Resultado del Tratamiento , Factor de Necrosis Tumoral alfa/análisis

RESUMEN

Major depression is a widespread, often chronic disorder affecting the individual, his or her family, and society as a whole. It incurs tremendous social and financial costs in the form of impaired relationships, lost productivity, and lost wages. Although chronic major depression is eminently treatable, it continues to be undertreated and underrecognized. This is particularly true in primary care settings, where physicians are usually the first to encounter chronic depression but are seldom trained to distinguish depression from other medical illnesses with similar symptoms. In addition, because of the stigma attached to depression, patients often characterize their symptoms as part of a physical illness or fail to report them to a clinician at all. This article discusses the epidemiology of depression, its impact and burden on society, and its special character (including diagnosis and treatment) as a chronic illness.

Asunto(s)

Costo de Enfermedad , Trastorno Depresivo/epidemiología , Enfermedad Crónica , Costos y Análisis de Costo , Atención a la Salud/normas , Atención a la Salud/estadística & datos numéricos , Atención a la Salud/tendencias , Trastorno Depresivo/economía , Trastorno Depresivo/terapia , Costos de la Atención en Salud/estadística & datos numéricos , Costos de la Atención en Salud/tendencias , Humanos , Prevalencia , Atención Primaria de Salud/normas , Atención Primaria de Salud/estadística & datos numéricos , Años de Vida Ajustados por Calidad de Vida , Recurrencia , Estereotipo , Estados Unidos/epidemiología

RESUMEN

OBJECTIVE: Infiltration of rheumatoid arthritis (RA) synovial tissue (ST) by differentiated dendritic cells (DC) is a consistent feature in patients with active disease. However, mononuclear cells (MNC), including DC, may be nonspecifically chemoattracted to inflammatory sites regardless of etiology. Therefore, to evaluate the specificity of ST infiltration by differentiated DC, synovial biopsies from patients with RA, spondylarthropathy (SpA), osteoarthritis (OA), and gout were examined. METHODS: Formalin-fixed ST sections were analyzed by double immunohistochemical staining for vascularity and infiltration by differentiated DC, lymphocytes, and macrophages. RESULTS: DC containing nuclear RelB were found in perivascular MNC aggregates from patients with all arthritides studied. Infiltration by differentiated DC was similar in RA and SpA ST, but reduced in OA ST. Differentiated DC were always observed in close association with lymphocytes, and the correlation between these variables suggested that the infiltration of inflammatory sites by DC and lymphocytes was associated. CONCLUSION: Perivascular infiltration by DC, lymphocytes, and macrophages is nonspecifically related to inflammation, but signals present in RA and SpA ST lead to more intense cellular infiltration and accumulation.

Asunto(s)

Enfermedades Autoinmunes/patología , Células Dendríticas/citología , Osteoartritis/patología , Membrana Sinovial/citología , Anciano , Anciano de 80 o más Años , Diferenciación Celular/fisiología , Femenino , Humanos , Artropatías/patología , Masculino , Persona de Mediana Edad , Enfermedades de la Columna Vertebral/patología

RESUMEN

In rheumatoid arthitis patients, three compartments need to be considered: peripheral blood (PB), synovial fluid (SF), and synovial tissue (ST). Dendritic cells (DC) characterized from each compartment have different properties. The methods given are based on cell sorting for isolation of cells, and flow cytometry and immunohistochemical staining for analysis of cells. Myeloid non-T cells are first enriched by density gradient centrifugation, sheep erythocyte rosetting, and, in some cases, magnetic immunodepletion. By flow cytometry, DC can then be analyzed or sorted based on two- or three-color immunofluorescence. Some variations on this basic theme are also outlined. The basic protocol for two-color immunohistochemistry of formalin-fixed ST is then given. This is based on the localization of the NFκB family member, RelB, to the nucleus of differentiated DC, and exclusion of B cells, macrophages, and follicular DC by double staining. Some variations that are particularly useful in frozen sections follow.

RESUMEN

Mice with targeted RelB mutations demonstrated an essential role for RelB in immune responses and in myeloid dendritic cell differentiation. Human studies suggested a more global transcriptional role in antigen presentation. Burkitt lymphoma cell lines were used as a model to examine the role of RelB in antigen presentation. After transient transfection of BJAB with RelB, strong nuclear expression of RelB-p50 heterodimers was associated with increased APC function and expression of CD40 and MHC class I. Antisense RelB in DG75 reduced antigen-presenting capacity and CD40-mediated up-regulation of MHC molecules. The data indicate that RelB transcriptional activity directly affects antigen presentation and CD40 synthesis. Stimulation of RelB transcriptional activity may provide a positive feedback loop for facilitating productive APC/T cell interactions.

Asunto(s)

Células Presentadoras de Antígenos/inmunología , Linfocitos B/inmunología , Núcleo Celular/metabolismo , Complejo Mayor de Histocompatibilidad , Proteínas Proto-Oncogénicas/metabolismo , Factores de Transcripción/metabolismo , Células Presentadoras de Antígenos/efectos de los fármacos , Línea Celular , Humanos , Oligonucleótidos Antisentido/farmacología , Transporte de Proteínas , Proteínas Proto-Oncogénicas/genética , Factor de Transcripción ReIB , Factores de Transcripción/genética