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This paper presents an approach based on triple injection capillary zone electrophoresis for identification of monoclonal antibodies. The analyte to be identified is injected between two zones of a known reference. The distances between the reference zones (plug I and III) and the target zone (plug II) are adjusted by partial electrophoresis of the first and second injection plugs. The full migration time of the target analyte is calculated from the observed migration time by considering the migration times of the reference in the first and third injection plugs. The relative migration time, that is, the ratio between the full migration time of the analyte and the migration time of the reference in the third injection plug provides the basis for identification. Here, eight monoclonal antibodies, including a pair of biosimilars, were used interchangeably as both analyte and reference to investigate potential of the method. The relative migration time for a preliminary positive identification were found to vary between 0.994 and 1.006 (1.000 ± 0.006, p = 95%). Beside the relative migration time, isoform distribution, peak profiles, and early migrating peaks, originating from components in the pharmaceutical formulations, were successfully used to verify the identity of all tested monoclonal antibodies.
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Anticorpos Monoclonais , Eletroforese Capilar , Anticorpos Monoclonais/análise , Anticorpos Monoclonais/químicaRESUMO
Academic health centers are faced with limited resources to navigate placement of advanced practice RN (APRN) students at the same time as APRN programs continue to increase. Clinical practicum sites are in higher demand due to clinical hour increases by accrediting organizations. This article describes a successful approach to the inefficient use of administration and preceptor resources for APRN student practicum placements via an innovative academic health system partnership.
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Bacharelado em Enfermagem , Estudantes de Enfermagem , Humanos , Universidades , Estudantes , Preceptoria , Comportamento CooperativoRESUMO
BACKGROUND: Significant growth in advanced practice registered nurse (APRN) student placements for clinical practicums have forced APRN preceptors, hospitals, and clinics to restrict the volume of APRN students they can accommodate. PURPOSE: Describe the experiences and opinions of APRNs regarding their precepting experiences of APRN students in three main areas; placement antecedents, placement postcedents, and preceptor incentives. METHODS: Quality improvement study conducted via electronic survey that contained quantitative short answer and a qualitative open-ended question of APRNs licensed in the State of REDACTED in June, 2019. FINDINGS: A total of 757 surveys were quantitatively analyzed. Qualitative thematic analysis resulted in identifying one overarching theme, the need for shared responsibility and accountability, and six unique primary themes. DISCUSSION: Our findings indicate that APRN preceptors recognize their critical role but lack support and preparation to fully execute academic and profession strategic goals and objectives. There is the need for academic and professional ownership of APRN education at every level.
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Prática Avançada de Enfermagem/educação , Atitude do Pessoal de Saúde , Educação de Pós-Graduação em Enfermagem/organização & administração , Preceptoria/organização & administração , Humanos , Melhoria de Qualidade , Inquéritos e Questionários , Estados UnidosRESUMO
BACKGROUND: Biologic' therapies, such as autologous conditioned serum (ACS), are gaining popularity in treating orthopaedic conditions in equine veterinary medicine. Evidence is scarce regarding ACS constituents, and large inter-individual differences in cytokine and growth factor content have been demonstrated. The objective of the current study was to investigate the potential association between cytokine and growth factor content of ACS and clinical effect in harness racehorses with spontaneously occurring low-grade articular lameness. Horses received 3 intra-articular injections of ACS administered at approximately 2-week intervals. Lameness evaluation consisting of a trot-up with subsequent flexions tests was performed at inclusion and approximately 2 weeks after the last treatment (re-evaluation); horses were classified as responders when there was no detectable lameness on trot-up and a minimum of 50% reduction in flexion test scores at re-evaluation. Association between clinical outcome (responders vs. non-responders) and age, lameness grades at inclusion (both initial trot-up and after flexion tests), treatment interval, follow-up time and the ACS content of IL-1Ra, IGF-1 and TGF-ß was determined by regression modelling. RESULTS: Outcome analysis was available for 19 of 20 included horses; 11 responded to treatment whereas 8 did not. There was considerable inter-individual variability in cytokine/growth factor content of ACS, and in the majority of the horses, the level of IL-10, IL-1ß and TNF-α was below the detection limit. In the final multivariate logistic regression model, ACS content of IGF-1 and IL-1Ra was significantly associated with clinical response (P = 0.01 and P = 0.03, respectively). No association with clinical response was found for the other tested variables. CONCLUSIONS: The therapeutic benefit of ACS may be related to higher levels of IL-1Ra and IGF-1. Our study corroborates previous findings of considerable inter-individual variability of cytokine- and growth factor content in ACS.
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Terapia Biológica/veterinária , Doenças dos Cavalos/terapia , Coxeadura Animal/terapia , Soro/química , Animais , Estudos de Coortes , Feminino , Cavalos , Injeções Intra-Articulares/veterinária , Fator de Crescimento Insulin-Like I/análise , Proteína Antagonista do Receptor de Interleucina 1/sangue , Masculino , Estudos Prospectivos , Fator de Crescimento Transformador beta/sangue , Resultado do TratamentoRESUMO
Collegial relationships, administrative champions, and persistence are key components to breaking down barriers to advanced practice RN (APRN) practice. This article addresses how Iowa APRNs in a state-sanctioned task force identified barriers for practicing at the top of their licensure in a full practice authority state including defending the right to control the scope of nursing practice in court.
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Prática Avançada de Enfermagem/organização & administração , Licenciamento em Enfermagem/normas , Profissionais de Enfermagem/organização & administração , Prática Avançada de Enfermagem/normas , Humanos , Iowa , Profissionais de Enfermagem/normasRESUMO
OBJECTIVE: This study examines data collected from a survey of advanced practice providers' (APPs') perceptions of reasonable versus actual APP-to-patient ratios and other factors that affect workload burden in both inpatient and outpatient clinical settings. BACKGROUND: Advanced practice providers provide accessible, cost-effective, and quality care in a growing number of specialty practices across multiple patient care settings. They are caring for higher volumes of patients and assuming more responsibilities while continuing to navigate highly complex healthcare systems. Limited evidence or benchmark data exist that would assist in determining optimal workload and staffing models that include APPs. METHODS: A group of advanced practice leaders developed and distributed a 43-question survey of workload factors to a highly diverse APP workforce. RESULTS: There were 1466 APPs across 37 areas of practice who responded to the survey distributed in 14 separate organizations. The perceived reasonable workload was lower than the actual workload for 22 specialty practice areas. The analysis included years of experience as an APP, work hours, on-call commitment, nonclinical responsibilities, and time spent in documentation, direct patient care, and performing procedures. CONCLUSIONS: There is a consensus among APPs, within their practice area, about what constitutes a reasonable patient assignment, despite the variability in APP experience, organizational culture, processes, and patient acuity.
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Prática Avançada de Enfermagem/organização & administração , Profissionais de Enfermagem/organização & administração , Papel do Profissional de Enfermagem/psicologia , Equipe de Assistência ao Paciente/organização & administração , Carga de Trabalho/psicologia , Humanos , Avaliação de Resultados em Cuidados de Saúde , Qualidade da Assistência à SaúdeRESUMO
The molecular aspects of inflammation were investigated in equine articular cartilage explants using quantitative proteomics. Articular cartilage explants were stimulated with interleukin (IL)-1ß in vitro for 25 days, and proteins released into cell culture media were chemically labeled with isobaric mass tags and analyzed by liquid chromatography-tandem mass spectrometry. A total of 127 proteins were identified and quantified in media from explants. IL-1ß-stimulation resulted in an abundance of proteins related to inflammation, including matrix metalloproteinases, acute phase proteins, complement components and IL-6. Extracellular matrix (ECM) molecules were released at different time points, and fragmentation of aggrecan and cartilage oligomeric matrix protein was observed at days 3 and 6, similar to early-stage OA in vivo. Degradation products of the collagenous network were observed at days 18 and 22, similar to late-stage OA. This model displays a longitudinal quantification of released molecules from the ECM of articular cartilage. Identification of dynamic changes of extracellular matrix molecules in the secretome of equine explants stimulated with IL-1ß over time may be useful for identifying components released at different time points during the spontaneous OA process.
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Cartilagem Articular/metabolismo , Matriz Extracelular/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Osteoartrite/metabolismo , Animais , Cartilagem Articular/patologia , Matriz Extracelular/patologia , Cavalos , Inflamação/metabolismo , Inflamação/patologia , Osteoartrite/patologiaRESUMO
Formation of synovial joints includes phenotypic changes of the chondrocytes and the organisation of their extracellular matrix is regulated by different factors and signalling pathways. Increased knowledge of the normal processes involved in joint development may be used to identify similar regulatory mechanisms during pathological conditions in the joint. Samples of the distal radius were collected from prenatal and postnatal equine growth plates, zones of Ranvier and articular cartilage with the aim of identifying Notch signalling components and cells with stem cell-like characteristics and to follow changes in matrix protein localisation during joint development. The localisation of the Notch signalling components Notch1, Delta4, Hes1, Notch dysregulating protein epidermal growth factor-like domain 7 (EGFL7), the stem cell-indicating factor Stro-1 and the matrix molecules cartilage oligomeric matrix protein (COMP), fibromodulin, matrilin-1 and chondroadherin were studied using immunohistochemistry. Spatial changes in protein localisations during cartilage maturation were observed for Notch signalling components and matrix molecules, with increased pericellular localisation indicating new synthesis and involvement of these proteins in the formation of the joint. However, it was not possible to characterise the phenotype of the chondrocytes based on their surrounding matrix during normal chondrogenesis. The zone of Ranvier was identified in all horses and characterised as an area expressing Stro-1, EGFL7 and chondroadherin with an absence of COMP and Notch signalling. Stro-1 was also present in cells close to the perichondrium, in the articular cartilage and in the fetal resting zone, indicating stem cell-like characteristics of these cells. The presence of stem cells in the articular cartilage will be of importance for the repair of damaged cartilage. Perivascular chondrocytes and hypertrophic cells of the cartilage bone interface displayed positive staining for EGFL7, which is a novel finding and suggests a role of EGFL7 in the vascular infiltration of growth cartilage.
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Biomarcadores/metabolismo , Cartilagem Articular/crescimento & desenvolvimento , Condrócitos/fisiologia , Lâmina de Crescimento/citologia , Cavalos/crescimento & desenvolvimento , Animais , Cartilagem Articular/citologia , Diferenciação Celular , Condrócitos/citologia , HomeostaseRESUMO
Changing demographics with an increased proportion of older adults indicate the need to develop new health-promoting interventions where the potential of digitization is considered. The aim was to explore and create an understanding of how social online meetings are experienced by older adults. Interviews with older adults generated data that were analyzed using a grounded theory approach. The participants were interviewed after participating in a digital health promotion group initiative provided in a municipality context. A core conceptual category and three subcategories reflected an intertwined process of discovering facets of social participation where internal reflections on personal values and needs were nurtured by an external driven process of becoming part of a group in an online context. Occupational therapists and other health and social care professionals need to consider the various facets of social participation when supporting older adults active and healthy aging.
Older Adults' Experiences of "Social Online Meetings"Promoting health among older adults is an important societal issue. Changing demographics with an increased proportion of older adults indicate the need to develop new health-promoting interventions. However, it is important to explore how such interventions in municipal settings meet the needs of older adults and the circumstances that influence their participation. Therefore, this study aimed to understand how older adults experience social online meetings. Seven older adults who had participated in an intervention called "social online meetings" were interviewed about their experiences. The findings demonstrate how older adults come to explore various dimensions of social participation and the value it holds for their everyday life. This gave rise to achieving new perspectives on everyday activities and encouraged personal development, and generated ideas for change. Occupational therapists and other health and social care professionals should consider how older adults' social participation can be promoted when designing health-promoting interventions online.
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BACKGROUND: Fostering social participation for active and healthy ageing among older adults is an urgent issue in a changing society that requires new approaches from occupational therapists as well as from society at large. AIM: To explore possibilities to foster social participation for older adults in society from the perspective of stakeholders. MATERIAL AND METHODS: A qualitative design was applied. 18 key informants, engaged in supporting older adults in their organisational roles as either professionals or volunteers, participated in five different focus groups discussions. RESULTS: The results involved two overarching themes that reflects different perspectives on key informants' possibilities to foster social participation for older adults. The first theme describes how they direct their attention towards their roles within the organisations to foster social participation in their own context. The second theme describes the broader societal perspectives that they address to establish a common ground for collaboration and knowledge-sharing among different stakeholders. CONCLUSIONS: The findings emphasise how addressing common challenges and developing collaboration are essential to foster older adults' social participation. It is therefore necessary to involve policy-makers and decision-makers. Occupational therapists and researchers should consider the value of occupational justice to drive collective and social approaches.
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BACKGROUND: Autologous conditioned serum (ACS) is used to treat osteoarthritis in horses, although its effects are not fully investigated. OBJECTIVES: To investigate the effects of equine serum and conditioned serum on chondrocytes stimulated with interleukin (IL)-1ß and cartilage explants with mild osteoarthritis. STUDY DESIGN: In vitro experimental study. METHODS: The effect of three different serum preparations (unincubated control [PS], serum incubated 24 h [PS24h] and serum incubated 24 h in ACS containers [PCS]) pooled from lame horses were tested in two in vitro models. IL-1ß and IL-1 receptor antagonist (IL-1Ra) concentrations were measured in all sera. In model 1, chondrocyte pellet cultures were stimulated with IL-1ß prior to treatment with the serum preparations for 2 and 48 h. Microarray, polymerase chain reaction, and matrix metallopeptidase-13 analyses were performed. In model 2, cartilage explants from horses with structural osteoarthritis were treated with PS or PCS on days 0, 6 and 12, or left untreated, and evaluated at day 24 using the OARSI grading scale for histological evaluation of articular cartilage. RESULTS: The IL-1Ra concentration in PS24h and PCS was significantly higher than in PS. In model 1, inflammation- and cartilage matrix degradation-related genes were upregulated after 48 h in all treatment groups versus untreated controls. Cartilage matrix molecules, aggrecan and collagens, were downregulated in PS24h- and PCS-treated pellets versus untreated controls. Growth factor signalling genes were upregulated-FGF7 in all treatment groups, BMP2 in PS24h-, and INHBA in PCS-treated-compared with untreated controls. In model 2, the OARSI score at day 24 was not significantly different between treatment groups. MAIN LIMITATIONS: Results from in vitro models cannot be directly translated to in vivo situations. CONCLUSIONS: In vitro treatment with conditioned serum did not alleviate IL-1ß-induced responses in chondrocyte pellets or lead to morphological improvement in osteoarthritic cartilage explants.
HISTORIAL: Suero autólogo acondicionado (ACS) es usado para tartar osteoartritis en caballos, aunque sus efectos no han sido completamente investigados. OBJETIVOS: Investigar los efectos de suero equino y suero acondicionado en condrocitos estimulados con interleukina (IL)-1ß y explantes de cartílago con osteoartritis leve. DISEÑO DEL ESTUDIO: Estudio experimental in vitro. MÉTODOS: El efecto de tres preparaciones séricas diferentes (control no incubado (PS), suero incubado 24 h (PS24h), y suero incubado 24 h en frascos ACS (PCS)) combinados y obtenidos de caballos cojos fueron probados en dos modelos in vitro. Las concentraciones de IL-1ß y de receptor antagonista de IL-1 (IL-1Ra) fueron medidas en todos los sueros. En el modelo 1, los cultivos de pellets de condrocitos fueron estimulados con IL-1ß antes de ser tratados con las preparaciones séricas durante 2 y 48 h. Se realizaron análisis de micromatrices, reacciones de polimerasa en cadena y de matriz de metalopeptidasa-13. En el modelo 2, explantaciones de cartílago proveniente de caballos con osteoartritis estructural fueron tratados con PS o PCS en los días 0, 6 y 12, o dejados sin tartar, y evaluados al día 24 usando la escala de graduación OARSI para evaluación histológica de cartílago articular. RESULTADOS: La concentración de IL-1Ra en PS24h y PCS fue significativamente mayor que en PS. En el modelo 1, los genes relacionados a la inflamación y a la degradación de la matriz cartilaginosa estaban aumentados después de 48 h en todos los grupos tratados en comparación a los controles no tratados. Las moléculas de matriz cartilaginosa, agrecanos y colágenos estaban disminuidos en los pellets PS24h y PCS versus los controles no tratados. Los genes de señales de factores de crecimiento FGF7 estaban aumentados en todos los grupos tratados, BMP2 en PS24h y INHBA in PCS en comparación con los controles no tratados. En el modelo 2, la escala OARSI al día 24 no fue significativamente distinta entre los grupos de tratamientos. LIMITACIONES PRINCIPALES: Los resultados de modelos in vitro no pueden ser directamente aplicados a situaciones in vivo. CONCLUSIONES: El tratamiento in vitro con suero acondicionado no alivió las respuestas inducidas por IL-1ß en pellets de condrocitos o llevo a mejoramiento morfológico en explantes de cartílago con osteoartritis.
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Cartilagem Articular , Doenças dos Cavalos , Osteoartrite , Cavalos , Animais , Condrócitos/metabolismo , Proteína Antagonista do Receptor de Interleucina 1/metabolismo , Proteína Antagonista do Receptor de Interleucina 1/farmacologia , Osteoartrite/terapia , Osteoartrite/veterinária , Inflamação/metabolismo , Inflamação/veterinária , Células Cultivadas , Doenças dos Cavalos/metabolismoRESUMO
BACKGROUND: Social participation concerns the possibility of engaging in activities in interaction with others or society at large and contributes to the health and well-being of older adults. In contrast, a lack of social participation is associated with loneliness. It is, therefore, important to understand what strategies older adults use to maintain social participation. AIM: To explore and describe older adults' experiences of maintaining social participation. MATERIALS AND METHODS: Nine older adults, aged 69-92 years, participated in interviews that were analysed through qualitative content analysis. RESULTS: The overall findings reflect how older adults create opportunities for social participation by developing strategies and striving to adapt to changing situations, emphasizing how maintaining social participation is an active process. Preserving social participation requires motivation and an effort to take initiative to cultivate social relations, maintain community bonds, engage in social events and activities and stay connected with society. CONCLUSIONS AND SIGNIFICANCE: These results may extend our understanding of strategies that older adults use, as well as the challenges they face when striving to adapt to new circumstances. This study may have implications for the practice of how to support older adults' social participation.
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Solidão , Participação Social , Idoso , Idoso de 80 Anos ou mais , Humanos , Solidão/psicologia , Motivação , Pesquisa Qualitativa , Participação Social/psicologiaRESUMO
BACKGROUND: Autologous conditioned serum (ACS) is used to treat osteoarthritis in horses, although its effects are not fully investigated. OBJECTIVES: To investigate the effects of equine serum and conditioned serum on chondrocytes stimulated with interleukin (IL)-1ß and cartilage explants with mild osteoarthritis. STUDY DESIGN: In vitro experimental study. METHODS: The effect of three different serum preparations (unincubated control [PS], serum incubated 24 h [PS24h], and serum incubated 24 h in ACS containers [PCS]) pooled from lame horses were tested in two in vitro models. IL-1ß and IL-1 receptor antagonist (IL-1Ra) concentrations were measured in all sera. In model 1, chondrocyte pellet cultures were stimulated with IL-1ß prior to treatment with the serum preparations for 2 and 48 h. Microarray, polymerase chain reaction, and matrix metallopeptidase-13 analyses were performed. In model 2, cartilage explants from horses with structural osteoarthritis were treated with PS or PCS on days 0, 6, and 12, or left untreated, and evaluated at day 24 using the OARSI grading scale for histological evaluation of articular cartilage. RESULTS: The IL-1Ra concentration in PS24h and PCS was significantly higher than in PS. In model 1, inflammation- and cartilage matrix degradation-related genes were upregulated after 48 h in all treatment groups versus untreated controls. Cartilage matrix molecules, aggrecan and collagens, were downregulated in PS24h- and PCS- treated pellets versus untreated controls. Growth factor signalling genes were upregulated-FGF7 in all treatment groups, BMP2 in PS24h-, and INHBA in PCS-treated- compared with untreated controls. In model 2, the OARSI score at day 24 was not significantly different between treatment groups. MAIN LIMITATIONS: Results from in vitro models cannot be directly translated to in vivo situations. CONCLUSIONS: In vitro treatment with conditioned serum did not alleviate IL-1ß-induced responses in chondrocyte pellets or lead to morphological improvement in osteoarthritic cartilage explants.
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This nation is facing a significant nursing faculty shortage not only at the undergraduate but also at the graduate level. As nursing faculty, we must be innovative in considering ways of consolidating resources in this time of demand for advanced practice nurses. With the apparent need for qualified neonatal nurse practitioners in Iowa, the University of Iowa understood the importance of starting a neonatal nurse practitioners program. The University of Iowa College of Nursing and the University of Missouri-Kansas City formed a partnership to educate neonatal nurse practitioners. Although this partnership concentrates on a specific subspecialty population, it can serve as a pilot project that could be adapted by those who care for other diverse populations. The nurse faculty shortage requires innovative solutions, and a model such as this one may provide solutions for other institutions.
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Docentes de Enfermagem/provisão & distribuição , Relações Interinstitucionais , Enfermagem Neonatal/educação , Profissionais de Enfermagem/educação , Escolas de Enfermagem/organização & administração , Humanos , Iowa , Missouri , Modelos Organizacionais , Desenvolvimento de ProgramasRESUMO
Mild, moderate and severe equine asthma is a problem for equine welfare. The aetiology of the disease is not known in detail but is likely multi-factorial. One important factor may be inhaled dust which carries harmful substances which may be bioactivated and thus can lead to local inflammation in the airways. The aim of this study was to investigate gene expression and protein localisation of cytochrome P450 (CYP) enzymes, superoxide dismutase and glutathione-S-transferases (GST) involved in bioactivation and detoxification of harmful substances in lungs of horses with or without histological evidence of lower airway inflammation. Significantly lower gene expression of CYP2A13 and GSTM1 was observed in lungs from horses with histological evidence of lower airway inflammation compared with horses without. A higher expression, although not significant, was found for CYP1A1 in horses with histological evidence of lower airway inflammation. There were no differences in gene expression of GSTP1 and SOD3. The proteins were localised in the respiratory epithelium which is of relevance as a defence to local exposure of inhaled harmful substances. In conclusion, our study reports differential gene expression of enzymes involved in bioactivation and detoxification of foreign substances in the lungs of horses with histological evidence of lower airway inflammation compared with horses without.
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Asma/veterinária , Doenças dos Cavalos/metabolismo , Inflamação/veterinária , Pulmão/enzimologia , Xenobióticos/metabolismo , Animais , Asma/imunologia , Asma/metabolismo , Feminino , Doenças dos Cavalos/imunologia , Cavalos , Inflamação/imunologia , Inflamação/metabolismo , Pulmão/fisiologia , MasculinoRESUMO
BACKGROUND: Dexamethasone is used for the intra-articular route of administration in management of aseptic arthritis in horses. Despite its widespread use there is very little quantitative data of the disposition and response to dexamethasone. The aim of this study was to investigate and describe the synovial fluid and plasma dexamethasone concentration over time and to explore the relation between synovial fluid concentration and response using clinical endpoints as response biomarkers after IA injection of dexamethasone disodium salt solution in an equine model of synovitis. RESULTS: Inflammation was induced in the radiocarpal joint of six horses by injection of 2 ng lipopolysaccharide (LPS). Two hours later either saline or dexamethasone was injected in the same joint in a two treatment cross over design. Each horse was treated once with one of the six doses dexamethasone used (0.01, 0.03, 0.1, 0.3, 1 or 3 mg) and once with saline. Dexamethasone was quantified by means of UHPLC-MS/MS. Dexamethasone disposition was characterised by means of a non-linear mixed effects model. Lameness was evaluated both objectively with an inertial sensor based system and subjectively scored using a numerical scale (0-5). Joint circumference, skin temperature over the joint and rectal temperature were also recorded. The LPS-challenge induced lameness in all horses with high inter-individual variability. Dexamethasone significantly decreased lameness compared with saline. Other variables were not statistically significant different between treatments. Objective lameness scoring was the most sensitive method used in this study to evaluate the lameness response. A pharmacokinetic/pharmacodynamic model was successfully fitted to experimental dexamethasone and lameness data. The model allowed characterization of the dexamethasone synovial fluid concentration-time course, the systemic exposure to dexamethasone after intra-articular administration and the concentration-response relation in an experimental model of synovitis. CONCLUSIONS: The quantitative data improve the understanding of the pharmacology of dexamethasone and might serve as input for future experiments and possibly contribute to maintain integrity of equine sports.
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Dexametasona/administração & dosagem , Doenças dos Cavalos/induzido quimicamente , Doenças dos Cavalos/tratamento farmacológico , Lipopolissacarídeos , Sinovite/veterinária , Animais , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/farmacocinética , Dexametasona/farmacocinética , Cavalos , Injeções Intra-Articulares/veterinária , Sinovite/induzido quimicamente , Sinovite/tratamento farmacológicoRESUMO
Osteoarthritis is an inflammatory and degenerative joint disease commonly affecting horses. To identify genes of relevance for cartilage pathology in osteoarthritis we studied the time-course effects of interleukin (IL)-1ß on equine articular cartilage. Articular cartilage explants from the distal third metacarpal bone were collected postmortem from three horses without evidence of joint disease. The explants were stimulated with IL-1ß for 27â¯days and global gene expression was measured by microarray. Gene expression was compared to that of unstimulated explants at days 3, 9, 15, 21 and 27. Release of inflammatory proteins was measured using Proximity Extension Assay. Stimulation with IL-1ß led to time-dependent changes in gene expression related to inflammation, the extracellular matrix (ECM), and phenotypic alterations. Gene expression and protein release of cytokines, chemokines, and matrix-degrading enzymes increased in the stimulated explants. Collagen type II was downregulated from day 15, whereas other ECM molecules were downregulated earlier. In contrast molecules involved in ECM signaling (perlecan, chondroitin sulfate proteoglycan 4, and syndecan 4) were upregulated. At the late time points, genes related to a chondrogenic phenotype were downregulated, and genes related to a hypertrophic phenotype were upregulated, suggesting a transition towards hypertrophy later in the culturing period. The data suggest that this in vitro model mimics time course events of in vivo inflammation in OA and it may be valuable as an in vitro tool to test treatments and to study disease mechanisms.
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Cartilagem Articular/metabolismo , Regulação da Expressão Gênica , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Animais , Células Cultivadas , Condrócitos , Matriz Extracelular , Cavalos , OsteoartriteRESUMO
OBJECTIVES: Usher syndrome is an inherited disorder that is characterized by hearing impairment (HI), retinitis pigmentosa, and in some cases vestibular dysfunction. Usher syndrome type IIa is caused by mutations in USH2A. HI in these patients is highly heterogeneous and the present study evaluates the effects of different types of USH2A mutations on the audiometric phenotype. Data from two large centres of expertise on Usher Syndrome in the Netherlands and Sweden were combined in order to create a large combined sample of patients to identify possible genotype-phenotype correlations. DESIGN: A retrospective study on HI in 110 patients (65 Dutch and 45 Swedish) genetically diagnosed with Usher syndrome type IIa. We used methods especially designed for characterizing and testing differences in audiological phenotype between patient subgroups. These methods included Age Related Typical Audiograms (ARTA) and a method to evaluate the difference in the degree of HI developed throughout life between subgroups. RESULTS: Cross-sectional linear regression analysis of last-visit audiograms for the best hearing ear demonstrated a gradual decline of hearing over decades. The congenital level of HI was in the range of 16-33 dB at 0.25-0.5 kHz, and in the range of 51-60 dB at 1-8 kHz. The annual threshold deterioration was in the range of 0.4-0.5 dB/year at 0.25-2 kHz and in the range of 0.7-0.8 dB/year at 4-8 kHz. Patients with two truncating mutations, including homozygotes for the common c.2299delG mutation, developed significantly more severe HI throughout life than patients with one truncating mutation combined with one nontruncating mutation, and patients with two nontruncating mutations. CONCLUSIONS: The results have direct implications for patient counselling in terms of prognosis of hearing and may serve as baseline measures for future (genetic) therapeutic interventions.