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The purpose of this prospective observational study was to determine whether dairy cattle housing types were associated with staphylococcal and mammaliicoccal populations found on teat skin, bedding, and in bulk tank milk. Twenty herds (n = 10 sand-bedded freestall herds; n = 10 compost-bedded pack herds) were enrolled. Each herd was visited twice for sample collection, and at each visit, 5 niches were sampled, including bulk tank milk, composite teat skin swab samples collected before premilking teat preparation, composite teat skin swab samples collected after premilking teat preparation, unused fresh bedding, and used bedding. All samples were plated on Mannitol salt agar and Columbia blood agar and staphylococcal-like colonies were selected for further evaluation. Bacterial colonies were speciated using MALDI-TOF mass spectrometry. All species were grouped into 4 categories included host-adapted, opportunistic, environmental, and unclassified. Absolute numbers and proportions of each genus and species were calculated. Proportional data were compared between groups using Fisher's exact test. Data representing 471 staphylococcal-like organisms were analyzed. Overall, 27 different staphylococcal and mammaliicoccal species were identified. Staphylococcus chromogenes was the only species identified from all 20 farms. A total of 20 different staphylococcal-like species were identified from bulk tank milk samples with the most prevalent species being S. chromogenes, followed by Staphylococcus aureus and Mammaliicoccus sciuri. Overall, more staphylococcal and mammaliicoccal isolates were identified among used bedding than unused bedding. The increased numbers of isolates within used bedding were primarily from used sand bedding samples, with 79% (76/96) of used bedding isolates being identified from sand bedding and only 20.8% (20/96) from used compost-bedded pack samples. When comparing categories found among sample types, more unclassified species were found in used sand bedding than in used compost-bedded pack samples. This finding is possibly related to the composting temperatures resulting in reduced growth or destruction of bacterial species. The prevalence of S. aureus was high in bulk tank milk for all herds, regardless of herd type, which may represent the influence of unmeasured management factors. Overall, staphylococcal and mammaliicoccal species were highly prevalent among samples from both farm types.
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Ropa de Cama y Ropa Blanca , Enfermedades de los Bovinos , Leche , Animales , Ropa de Cama y Ropa Blanca/veterinaria , Bovinos , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/microbiología , Compostaje , Industria Lechera , Granjas , Femenino , Vivienda para Animales , Leche/microbiología , Arena , Infecciones Estafilocócicas/epidemiología , Infecciones Estafilocócicas/veterinaria , Staphylococcus aureus/aislamiento & purificaciónRESUMEN
Predicting dry matter intake (DMI) and feed efficiency by leveraging the use of data streams available on farm could aid efforts to improve the feed efficiency of dairy cattle. Residual feed intake (RFI) is the difference between predicted and observed feed intake after accounting for body size, body weight change, and milk production, making it a valuable metric for feed efficiency research. Our objective was to develop and evaluate DMI and RFI prediction models using multiple linear regression (MLR), partial least squares regression, artificial neural networks, and stacked ensembles using different combinations of cow descriptive, performance, sensor-derived behavioral (SMARTBOW; Zoetis), and blood metabolite data. Data were collected from mid-lactation Holstein cows (n = 124; 102 multiparous, 22 primiparous) split equally between 2 replicates of 45-d duration with ad libitum access to feed. Within each predictive approach, 4 data streams were added in sequence: dataset M (week of lactation, parity, milk yield, and milk components), dataset MB (dataset M plus body condition score and metabolic body weight), dataset MBS (dataset MB plus sensor-derived behavioral variables), and dataset MBSP (dataset MBS plus physiological blood metabolites). The combination of 4 datasets and 4 analytical approaches resulted in 16 analyses of DMI and RFI, using variables averaged within cow across the study period. Additional models using weekly averaged data within cow and study were built using all predictive approaches for datasets M, MB, and MBS. Model performance was assessed using the coefficient of determination, concordance correlation coefficient, and root mean square error of prediction. Predictive models of DMI performed similarly across all approaches, and models using dataset MBS had the greatest model performance. The best approach-dataset combination was MLR-dataset MBS, although several models performed similarly. Weekly DMI models had the greatest performance with MLR and partial least squares regression approaches. Dataset MBS models had incrementally better performance than datasets MB and M. Within each approach-dataset combination, models with DMI averaged over the study period had slightly greater model performance than DMI averaged weekly. Predictive performance of all RFI models was poor, but slight improvements when using MLR applied to dataset MBS suggest that rumination and activity behaviors may explain some of the variation in RFI. Overall, similar performance of MLR, compared with machine learning techniques, indicates MLR may be sufficient to predict DMI. The improvement in model performance with each additional data stream supports the idea of integrating data streams to improve model predictions and farm management decisions.
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Alimentación Animal , Lactancia , Alimentación Animal/análisis , Animales , Peso Corporal , Bovinos , Dieta/veterinaria , Ingestión de Alimentos , Femenino , Leche , EmbarazoRESUMEN
Validation of precision dairy-monitoring technologies establishes technology behavioral-monitoring efficacy for research and commercial application. Technology metrics should be associated with behaviors of known physiological importance. The objective of this research project was to evaluate the Nedap SmartTag Neck (Nedap Livestock Management, Groenlo, the Netherlands) for dairy cow behavior measuring accuracy. The behaviors measured were eating, ruminating, and inactivity. Thirty-six lactating Holstein dairy cows were randomly selected from the University of Kentucky's Coldstream Dairy Research Herd and fitted with a Nedap SmartTag Neck. Cows were observed by a single observer for a total of 4â¯h per cow, including 2â¯h after the morning milking (0800â¯h) and 2â¯h after the evening milking (2000â¯h), from May to December 2017. The observer recorded the time behaviors occurred using a synchronized watch (CASIO, CASIO America, Inc., Dover, NJ, USA). The hour, minute, and second of the day each behavior occurred were compared with corresponding technology measurements. Pearson correlation coefficients (r; CORR procedure; SAS Institute Inc., Cary, NC, USA), concordance correlation coefficients (CCC; epiR package; R Foundation for Statistical Computing, Vienna, Austria), and Bland-Altman plots (epiR package; R Foundation for Statistical Computing) were used to determine association between visual observations and technology-recorded behaviors. Visually recorded eating, ruminating, and inactive time were moderately to strongly correlated with technology data (CCCâ¯≥â¯0.88) and Bland-Altman plots showed no bias, indicating a high level of agreement. In conclusion, the Nedap SmartTag Neck accurately monitored eating, ruminating, and inactivity behaviors and is expected to be effective in monitoring these behaviors in lactating dairy cattle in research or commercial farm settings.
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Conducta Alimentaria , Lactancia , Animales , Austria , Conducta Animal , Bovinos , Industria Lechera , Ingestión de Alimentos , Femenino , Países BajosRESUMEN
A 2-year-old male Djungarian hamster (Phodopus sungorus) and a 1-year-old male Roborovski hamster (Phodopus roborovskii) were presented with expansile subcutaneous masses. The well-demarcated, firm, grey, multilobulated nodules displayed an homogeneous, white-grey cut surface. Histological examination revealed a neoplasm of variable cellularity consisting of spindle-shaped to polygonal 'ganglion cell-like' cells with abundant, amphophilic, vacuolated cytoplasm. Immunohistochemically, the tumour cells were labelled intensely for vimentin and nestin, moderately for neuron specific enolase and weakly for melan-A. The histological and immunohistochemical findings were suggestive of an atypical fibroma with evidence of a neuroectodermal phenotype.
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Fibroma/veterinaria , Phodopus , Enfermedades de los Roedores/patología , Animales , Biomarcadores/metabolismo , Cricetinae , Fibroma/metabolismo , Inmunohistoquímica , Antígeno MART-1/metabolismo , Masculino , Nestina/metabolismo , Placa Neural/metabolismo , Fosfopiruvato Hidratasa/metabolismo , Vimentina/metabolismoRESUMEN
Precision dairy monitoring technologies have become increasingly popular for recording rumination and feeding behaviors in dairy cattle. The objective of this study was to validate the rumination and feeding time functions of the CowManager SensOor (Agis, Harmelen, the Netherlands) against visual observation in dairy heifers. The study took place over a 44-d period beginning June 1, 2016. Holstein heifers equipped with CowManager SensOor tags attached according to manufacturer specifications (n = 49) were split into 2 groups based on age, diet, and housing type. Group 1 heifers (n = 24) were calves (mean ± SD) 2.0 ± 2.7 mo in age, fed hay and calf starter, and housed on a straw-bedded pack. Group 2 heifers (n = 25) were 17.0 ± 1.3 mo in age, fed a TMR, confirmed pregnant, and housed in freestalls. Visual observation shifts occurred at 1500, 1700, 1900, and 2100 h. Each heifer was observed for 2 hour-long periods, with both observation periods occurring on the same day. Visual observations were collected using a synchronized watch, and "start" and "stop" times were recorded for each rumination and feeding event. For correlations, data from CowManager SensOor tags and observations were averaged, so a single 1-h observation was provided per animal, reducing the potential for confounding repeated measures being collected for each animal. Concordance correlations (CCC; epiR package; R Foundation for Statistical Computing, Vienna, Austria) and Pearson correlations (r; CORR procedure; SAS Institute Inc., Cary, NC) were used to calculate association between visual observations and technology-recorded behaviors. Visually observed rumination time was correlated with the CowManager SensOor (r = 0.63, CCC = 0.55). Visually observed feeding time was also correlated with the CowManager SensOor (r = 0.88, CCC = 0.72). The difference between technology-recorded data and visual observation was treated as the dependent variable in a mixed linear model (MIXED procedure of SAS). Time of day, age in months, and group were treated as fixed effects. Individual heifers were treated as random and repeated effects. The effects of time of day, age, and group on rumination and feeding times were not significant. The CowManager SensOor was more effective at recording feeding behavior than rumination behavior in dairy heifers. The CowManager SensOor can be used to provide relatively accurate measures of feeding time in heifers, but its rumination time function should be used with caution.
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Bovinos/fisiología , Conducta Alimentaria , Rumen/metabolismo , Alimentación Animal/análisis , Animales , FemeninoRESUMEN
The objective of this study was to use automated activity, lying, and rumination monitors to characterize prepartum behavior and predict calving in dairy cattle. Data were collected from 20 primiparous and 33 multiparous Holstein dairy cattle from September 2011 to May 2013 at the University of Kentucky Coldstream Dairy. The HR Tag (SCR Engineers Ltd., Netanya, Israel) automatically collected neck activity and rumination data in 2-h increments. The IceQube (IceRobotics Ltd., South Queensferry, United Kingdom) automatically collected number of steps, lying time, standing time, number of transitions from standing to lying (lying bouts), and total motion, summed in 15-min increments. IceQube data were summed in 2-h increments to match HR Tag data. All behavioral data were collected for 14 d before the predicted calving date. Retrospective data analysis was performed using mixed linear models to examine behavioral changes by day in the 14 d before calving. Bihourly behavioral differences from baseline values over the 14 d before calving were also evaluated using mixed linear models. Changes in daily rumination time, total motion, lying time, and lying bouts occurred in the 14 d before calving. In the bihourly analysis, extreme values for all behaviors occurred in the final 24 h, indicating that the monitored behaviors may be useful in calving prediction. To determine whether technologies were useful at predicting calving, random forest, linear discriminant analysis, and neural network machine-learning techniques were constructed and implemented using R version 3.1.0 (R Foundation for Statistical Computing, Vienna, Austria). These methods were used on variables from each technology and all combined variables from both technologies. A neural network analysis that combined variables from both technologies at the daily level yielded 100.0% sensitivity and 86.8% specificity. A neural network analysis that combined variables from both technologies in bihourly increments was used to identify 2-h periods in the 8 h before calving with 82.8% sensitivity and 80.4% specificity. Changes in behavior and machine-learning alerts indicate that commercially marketed behavioral monitors may have calving prediction potential.
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Actigrafía/veterinaria , Conducta Animal , Aprendizaje Automático , Masticación , Parto , Actigrafía/instrumentación , Actigrafía/métodos , Animales , Austria , Bovinos , Femenino , Israel , Postura/fisiología , Estudios Retrospectivos , Factores de Tiempo , Reino UnidoRESUMEN
The objective of this study was to evaluate commercially available precision dairy technologies against direct visual observations of feeding, rumination, and lying behaviors. Primiparous (n=24) and multiparous (n=24) lactating Holstein dairy cattle (mean ± standard deviation; 223.4±117.8 d in milk, producing 29.2±8.2kg of milk/d) were fitted with 6 different triaxial accelerometer technologies evaluating cow behaviors at or before freshening. The AfiAct Pedometer Plus (Afimilk, Kibbutz Afikim, Israel) was used to monitor lying time. The CowManager SensOor (Agis, Harmelen, Netherlands) monitored rumination and feeding time. The HOBO Data Logger (HOBO Pendant G Acceleration Data Logger, Onset Computer Corp., Pocasset, MA) monitored lying time. The CowAlert IceQube (IceRobotics Ltd., Edinburgh, Scotland) monitored lying time. The Smartbow (Smartbow GmbH, Jutogasse, Austria) monitored rumination time. The Track A Cow (ENGS, Rosh Pina, Israel) monitored lying time and time spent around feeding areas for the calculation of feeding time. Over 8 d, 6 cows per day were visually observed for feeding, rumination, and lying behaviors for 2 h after morning and evening milking. The time of day was recorded when each behavior began and ended. These times were used to generate the length of time behaviors were visually observed. Pearson correlations (r; calculated using the CORR procedure of SAS Version 9.3, SAS Institute Inc., Cary, NC), and concordance correlations (CCC; calculated using the epiR package of R version 3.1.0, R Foundation for Statistical Computing, Vienna, Austria) evaluated association between visual observations and technology-recorded behaviors. Visually recorded feeding behaviors were moderately correlated with the CowManager SensOor (r=0.88, CCC=0.82) and Track A Cow (r=0.93, CCC=0.79) monitors. Visually recorded rumination behaviors were strongly correlated with the Smartbow (r=0.97, CCC=0.96), and weakly correlated with the CowManager SensOor (r=0.69, CCC=0.59). Visually recorded lying behaviors were strongly correlated with the AfiAct Pedometer Plus (r >0.99, CCC >0.99), CowAlert IceQube (r >0.99, CCC >0.99), and Track A Cow (r >0.99, CCC >0.99). The HOBO Data Loggers were moderately correlated (r >0.83, CCC >0.81) with visual observations. Based on these results, the evaluated precision dairy monitoring technologies accurately monitored dairy cattle behavior.
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Conducta Animal , Lactancia , Animales , Bovinos , Industria Lechera , Ingestión de Alimentos , Conducta Alimentaria , Femenino , LecheRESUMEN
Recent progress in genetic testing has facilitated obtaining an etiologic diagnosis in children with developmental delay/intellectual disability (DD/ID) or multiple congenital anomalies (MCA) or both. Little is known about the benefits of diagnostic elucidation for affected families. We studied the impact of a genetic diagnosis on parental quality of life (QoL) using a validated semiquantitative questionnaire in families with a disabled child investigated by array-based comparative genomic hybridization (aCGH). We received completed questionnaires from 95 mothers and 76 fathers of 99 families. We used multivariate analysis for adjustment of potential confounders. Taken all 99 families together, maternal QoL score (percentile rank scale 51.05) was significantly lower than fathers' QoL (61.83, p = 0.01). Maternal QoL score was 20.17 [95% CI (5.49; 34.82)] percentile rank scales higher in mothers of children with diagnostic (n = 34) aCGH as opposed to mothers of children with inconclusive (n = 65) aCGH (Hedges' g = 0.71). Comparison of these QoL scores with retrospectively recalled QoL before aCGH revealed an increase of maternal QoL after diagnostic clarification. Our results indicate a benefit for maternal QoL if a genetic test, here aCGH, succeeds to clarify the etiologic diagnosis in a disabled child.
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Discapacidades del Desarrollo/diagnóstico , Discapacidades del Desarrollo/genética , Padres , Calidad de Vida , Adulto , Niño , Hibridación Genómica Comparativa , Factores de Confusión Epidemiológicos , Demografía , Femenino , Humanos , MasculinoRESUMEN
An online survey to identify producer precision dairy farming technology perception was distributed in March 2013 through web links sent to dairy producers through written publications and e-mail. Responses were collected in May 2013 and 109 surveys were used in statistical analysis. Producers were asked to select parameters monitored by technologies on their farm from a predetermined list and 68.8% of respondents indicated technology use on their dairies (31.2% of producers not using technologies). Daily milk yield (52.3%), cow activity (41.3%), and mastitis (25.7%) were selected most frequently. Producers were also asked to score the same list of parameters on usefulness using a 5-point scale (1=not useful and 5=useful). Producers indicated (mean ± SE) mastitis (4.77±0.47), standing estrus (4.75±0.55), and daily milk yield (4.72±0.62) to be most useful. Producers were asked to score considerations taken before deciding to purchase a precision dairy farming technology from a predetermined list (1=not important and 5=important). Producers indicated benefit-to-cost ratio (4.57±0.66), total investment cost (4.28±0.83), and simplicity and ease of use (4.26±0.75) to be most important when deciding whether to implement a technology. Producers were categorized based on technology use (using technology vs. not using technology) and differed significantly across technology usefulness scores, daily milk yield (using technologies: 4.83±0.07 vs. not using technologies: 4.50±0.10), and standing estrus (using technologies: 4.68±0.06 vs. not using technologies: 4.91±0.09). The same categories were used to evaluate technology use effect on prepurchase technology selection criteria and availability of local support (using technologies: 4.25±0.11 vs. not using technologies: 3.82±0.16) differed significantly. Producer perception of technology remains relatively unknown to manufacturers. Using this data, technology manufacturers may better design and market technologies to producer need.
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Bovinos , Industria Lechera/métodos , Tecnología/métodos , Animales , Industria Lechera/economía , Industria Lechera/instrumentación , Percepción , Tecnología/economía , Tecnología/instrumentaciónRESUMEN
Array comparative genomic hybridization (array CGH) is now widely adopted as a first-tier clinical diagnostic test in individuals with unexplained developmental delay/intellectual disability (DD/ID) and congenital anomalies. Our study aimed at enlarging the phenotypic spectrum associated with clinically relevant copy number variants (CNVs) as well as delineating clinical criteria, which may help separating patients with pathogenic CNVs from those without pathogenic CNVs. We performed a retrospective review of clinical and array CGH data of 342 children with unexplained DD/ID. The phenotypic features of patients with clinically significant CNV were compared with those without pathogenic CNVs. Array CGH detected pathogenic CNVs in 13.2% of the patients. Congenital anomalies, especially heart defects, as well as primary microcephaly, short stature and failure to thrive were clearly more frequent in children with pathogenic CNVs compared with children with normal array CGH results. Thus, we assume that in patients with unexplained DD/ID, array CGH will more probably detect a significant CNV if any of these features is part of the patient's phenotype.
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Hibridación Genómica Comparativa/métodos , Variaciones en el Número de Copia de ADN/genética , Discapacidades del Desarrollo , Discapacidad Intelectual , Adolescente , Niño , Discapacidades del Desarrollo/diagnóstico , Discapacidades del Desarrollo/genética , Insuficiencia de Crecimiento/genética , Insuficiencia de Crecimiento/fisiopatología , Femenino , Humanos , Lactante , Discapacidad Intelectual/genética , Discapacidad Intelectual/fisiopatología , Masculino , Microcefalia/genética , Microcefalia/fisiopatología , Fenotipo , Estudios RetrospectivosRESUMEN
OBJECTIVE: New teaching methods such as e-learning, are increasingly used to support common methods such as lectures, seminars and practical training in universities providing education in veterinary medicine. In the current study, the acceptance of e-learning in the example of the CASUS system by veterinarians as well as students of veterinary medicine of all German-speaking universities was analyzed. Material und methods: For this purpose an online evaluation questionnaire was developed. Members of the target groups were informed by e-mail and references in professional journals, as well as through veterinarian exchange platforms on the internet. Additionally, 224 students' final anatomy marks were compared and correlated to the utilization of CASUS to gain an important insight for the development of new teaching practices in the teaching of veterinary medicine. RESULTS: In total 1581 questionnaires were evaluated. A good acceptance regarding new teaching practices was found, although the classical textbook is still the most important instrument for imparting knowledge. The degree of utilization of e-learning strongly depends on its integration into the teaching content. CASUS is regarded as an efficient teaching method, with over 90% of the respondents indicating a strong desire to expand the number of case studies. Due to the present low degree of integration into the teaching content, no significant correlation could be found between the utilization of anatomy case studies and the final anatomy mark. However, based on their subjective perception, the students reported a high level of success in their study results with the likely effect of supporting increasing self-assurance in the situation of examinations. CONCLUSION: With the help of e-learning, educational objectives can be achieved that are not attainable by traditional teaching methods, e.g. the review of individual improvements by using the integrated feedback-function of e-learning programs. However, e-learning is not able to completely replace current teaching practices and hence should be considered as an additional element in future teaching models.
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HISTORY AND ADMISSION FINDINGS: A 71-year-old woman presented with suspected tuberculosis. She reported having productive coughs, unwanted weight loss and subfebrile temperature in the preceding 3 months. She was known to have chronic obstructive pulmonary disease treated with corticoids given systemically and by inhalation. She was a heavy smoker. INVESTIGATIONS: Computed tomography revealed a left apical lung abscess. In the further course of the disease magnetic resonance imaging of the head demonstrated multiple abscesses in both cerebral hemispheres and an abscess, 3.4 cm in diameter, in the right side of the cerebellum, as well as a intra-orbital tumor on the right. Needle aspirate of the eyeball grew Nocardia farcinica. TREATMENT AND COURSE: Over 3 weeks antimicrobial treatment was given with imipenem and amikacin, followed by oral cotrimoxazole for 12 months. The abscesses completely regressed and after 12 months no recurrence was demonstrated either radiologically or clinically. CONCLUSION: Although nocardiasis is rare in Germany it must be included in the differential diagnosis of pneumonia with abscesses. This is especially so if acid-fast bacilli are found. As the resistance pattern of N. farcinica to antibiotics varies, early treatment is essential with antibiotics to which it is sensitive.
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Absceso Encefálico/microbiología , Enfermedades Cerebelosas/microbiología , Absceso Pulmonar/microbiología , Nocardiosis/complicaciones , Enfermedades Orbitales/microbiología , Neumonía Bacteriana/complicaciones , Anciano , Amicacina/uso terapéutico , Antibacterianos/uso terapéutico , Biopsia con Aguja/métodos , Absceso Encefálico/tratamiento farmacológico , Absceso Encefálico/patología , Enfermedades Cerebelosas/tratamiento farmacológico , Enfermedades Cerebelosas/patología , Diagnóstico Diferencial , Quimioterapia Combinada , Femenino , Humanos , Imipenem/uso terapéutico , Absceso Pulmonar/tratamiento farmacológico , Absceso Pulmonar/patología , Imagen por Resonancia Magnética , Nocardiosis/diagnóstico , Nocardiosis/tratamiento farmacológico , Enfermedades Orbitales/tratamiento farmacológico , Enfermedades Orbitales/patología , Neumonía Bacteriana/diagnóstico , Neumonía Bacteriana/tratamiento farmacológico , Tomografía Computarizada por Rayos X , Resultado del Tratamiento , Combinación Trimetoprim y Sulfametoxazol/uso terapéuticoRESUMEN
OBJECTIVE: To compare the content covered by twelve obesity-specific health status measures using the International Classification of Functioning, Disability and Health (ICF). DESIGN: Obesity-specific health status measures were identified and then linked to the ICF separately by two trained health professionals according to standardized guidelines. The degree of agreement between health professionals was calculated by means of the kappa (kappa) statistic. Bootstrapped confidence intervals (CI) were calculated. The obesity-specific health-status measures were compared on the component and category level of the ICF. MEASUREMENTS: welve condition-specific health-status measures were identified and included in this study, namely the obesity-related problem scale, the obesity eating problems scale, the obesity-related coping and obesity-related distress questionnaire, the impact of weight on quality of life questionnaire (short version), the health-related quality of life questionnaire, the obesity adjustment survey (short form), the short specific quality of life scale, the obesity-related well-being questionnaire, the bariatric analysis and reporting outcome system, the bariatric quality of life index, the obesity and weight loss quality of life questionnaire and the weight-related symptom measure. RESULTS: In the 280 items of the eight measures, a total of 413 concepts were identified and linked to the 87 different ICF categories. The measures varied strongly in the number of concepts contained and the number of ICF categories used to map these concepts. Items on body functions varied form 12% in the obesity-related problem scale to 95% in the weight-related symptom measure. The estimated kappa coefficients ranged between 0.79 (CI: 0.72, 0.86) at the component ICFs level and 0.97 (CI: 0.93, 1.0) at the third ICF's level. CONCLUSION: The ICF proved highly useful for the content comparison of obesity-specific health-status measures. The results may provide clinicians and researchers with new insights when selecting health-status measures for clinical studies in obesity.
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Indicadores de Salud , Obesidad/rehabilitación , Adaptación Psicológica , Evaluación de la Discapacidad , Humanos , Obesidad/psicología , Psicometría , Calidad de Vida , Reproducibilidad de los ResultadosRESUMEN
UNLABELLED: Intracellular 1,25-dihydroxyvitamin D receptor (VDR) is expressed in human skeletal muscle tissue. However, it is unknown whether VDR expression in vivo is related to age or vitamin D status, or whether VDR expression differs between skeletal muscle groups. INTRODUCTION: We investigated these factors and their relation to 1,25-dihydroxyvitamin D receptor (VDR) expression in freshly removed human muscle tissue. MATERIALS AND METHODS: We investigated biopsy specimens of the gluteus medius taken at surgery from 20 female patients undergoing total hip arthroplasty (mean age, 71.6 +/- 14.5; 72% > 65 years) and biopsy specimens of the transversospinalis muscle taken at surgery from 12 female patients with spinal operations (mean age, 55.2 +/- 19.6; 28% > 65 years). The specimens were obtained by immunohistological staining of the VDR using a monoclonal rat antibody to the VDR (Clone no. 9A7). Quantitative VDR expression (number of VDR positive nuclei) was assessed by counting 500 nuclei per specimen and person. Serum concentrations of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D were assessed at day of admission to surgery. RESULTS: All muscle biopsy specimens stained positive for VDR. In the univariate analyses, increased age was associated with decreased VDR expression (r = 0.5: p = 0.004), whereas there were no significant correlations between VDR expression and 25-hydroxyvitamin D or 1,25-dihydroxyvitamin D levels. VDR expression did not differ between patients with hip and spinal surgery. In the multivariate analysis, older age was a significant predictor of decreased VDR expression after controlling biopsy location (gluteus medius or the transversospinalis muscle), and 25-hydroxyvitamin D levels (linear regression analysis: beta-estimate = -2.56; p = 0.047). CONCLUSIONS: Intranuclear immunostaining of the VDR was present in muscle biopsy specimens of all orthopedic patients. Older age was significantly associated with decreased VDR expression, independent of biopsy location and serum 25-hydroxyvitamin D levels.
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Envejecimiento/metabolismo , Músculo Esquelético/metabolismo , Receptores de Calcitriol/metabolismo , Adulto , Anciano , Biopsia , Femenino , Humanos , Persona de Mediana Edad , Músculo Esquelético/patologíaRESUMEN
The potential role of airway interleukin-5 (IL-5) expression in eliciting mucus production was demonstrated in a pulmonary IL-5 transgenic mouse model (NJ.1726) in which naive transgenic mice display comparable levels of airway mucus relative to allergen-sensitized and -challenged wild-type mice. The intrinsic mucus accumulation of NJ.1726 was abolished in compound transgenic-gene knockout mice deficient of either CD4(+) cells [NJ.1726/CD4(-/-)] or alphabeta T cell receptor-positive (TCR(+)) cells [NJ.1726/alphabeta TCR(-/-)]. In addition, mucus production in naive NJ.1726 was inhibited by >90% after administration of the soluble anti-IL-4 receptor alpha-subunit antagonist. The loss of mucus production in NJ.1726/CD4(-/-), NJ.1726/alphabeta TCR(-/-), and anti-IL-4 receptor alpha-subunit antagonist-treated mice occurred notwithstanding the significant pulmonary eosinophilia and expansion of airway B cells induced by ectopic IL-5 expression. Furthermore, the loss of mucus accumulation occurred in these mice despite elevated levels of airway and peripheral IL-5, indicating that IL-5 does not directly induce goblet cell metaplasia and mucus production. Thus pulmonary expression of IL-5 alone is capable of inducing CD4(+) T cell-dependent goblet cell metaplasia, apparently mediated by IL-4 receptor alpha-subunit-ligand interactions, and represents a previously unrecognized novel pathway for augmenting allergen-induced mucus production.
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Asma/patología , Linfocitos T CD4-Positivos/metabolismo , Interleucina-5/genética , Moco/metabolismo , Alérgenos/farmacología , Animales , Asma/inmunología , Asma/metabolismo , Eosinófilos/inmunología , Expresión Génica/inmunología , Células Caliciformes/metabolismo , Interleucina-5/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Moco/inmunología , Ovalbúmina/inmunología , Mucosa Respiratoria/metabolismoRESUMEN
Allergen-induced recruitment of T lymphocytes and eosinophils to the airways is associated with increased expression of the transcription factor GATA-3. In this study, the relationship between airway inflammation and GATA-3 expression in the lungs was investigated using ragweed-sensitized C57BL/6J mice. Intratracheal ragweed challenge increased both the number of GATA-3-expressing cells in the perivascular and peribronchial regions and the amount of expression per cell. Interleukin (IL)-4 and IL-5 levels in bronchoalveolar lavage fluid were upregulated in parallel with GATA-3 expression. GATA-3 mRNA and protein colocalized to eosinophils. Eosinophils isolated from the lungs and stimulated with phorbol 12-myristate 13-acetate and/or A-23187 released IL-5. The release was inhibited by actinomycin D, which indicates that de novo synthesis of the cytokine was involved. Western blot analysis of proteins from isolated eosinophils demonstrated expression of the p50 subunit of nuclear factor-kappaB, a transcription factor that is implicated in control of GATA-3 expression. These data provide evidence that allergen challenge increases GATA-3 and proinflammatory cytokine expression by pulmonary eosinophils, which could provide positive feedback for the inflammatory response.
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Proteínas de Unión al ADN/genética , Eosinófilos/inmunología , Interleucina-4/análisis , Interleucina-5/análisis , Hipersensibilidad Respiratoria/inmunología , Ribonucleasas , Transactivadores/genética , Alérgenos/inmunología , Animales , Asma/inmunología , Proteínas Sanguíneas/análisis , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/inmunología , Quimiotaxis de Leucocito/inmunología , Proteínas de Unión al ADN/análisis , Proteínas en los Gránulos del Eosinófilo , Eosinófilos/química , Eosinófilos/citología , Factor de Transcripción GATA3 , Expresión Génica/inmunología , Hibridación in Situ , Técnicas In Vitro , Interleucina-4/biosíntesis , Interleucina-5/biosíntesis , Masculino , Ratones , Ratones Endogámicos C57BL , Proteínas de Plantas/inmunología , ARN Mensajero/análisis , Células Th2/inmunología , Transactivadores/análisis , Transcripción Genética/inmunologíaRESUMEN
CD4(+) T cells have a critical role in the development of allergic pulmonary inflammation, including the recruitment of eosinophils to the airway lumen and interstitium. The expression of interleukin (IL)-5 by CD4(+) cells has, in particular, often been lionized as the central link between allergic inflammation and the concomitant expansion or recruitment of eosinophils. The mechanism(s) by which CD4(+) T cells mediates eosinophil recruitment was assessed with gene knockout mice deficient for T cells or T cell subtypes and a unique IL-5 transgenic mouse (line NJ.1726) that constitutively overexpresses this cytokine in the lung epithelium. Pulmonary IL-5 expression is significantly attenuated in T cell- and CD4(+) but not CD8(+) cell-deficient animals, suggesting an obvious explanation for the lack of eosinophils in the lungs of T cell-deficient and CD4(-/-) mice. However, although the constitutive expression of IL-5 in the lung epithelium of NJ.1726 mice elicited an eosinophilia in the airway lumen of both naive and ovalbumin-treated mice, in the absence of CD4(+) cells, allergen-mediated eosinophil recruitment to the bronchoalveolar lavage fluid was abolished. Moreover, intranasal instillation of the potent eosinophil-specific chemokine eotaxin-2 was incapable of eliciting eosinophil recruitment in naive and ovalbumin-treated NJ.1726 CD4(-/-) mice, suggesting that eosinophil trafficking during allergic inflammatory responses is a consequence of a CD4(+) cell-mediated event(s) in addition to IL-5 expression and the establishment of a pulmonary chemokine gradient.
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Linfocitos T CD4-Positivos/fisiología , Interleucina-5/metabolismo , Administración Intranasal , Alérgenos/inmunología , Animales , Quimiocina CCL24 , Quimiocinas CC/administración & dosificación , Quimiocinas CC/farmacología , Quimiotaxis de Leucocito/fisiología , Eosinófilos/efectos de los fármacos , Eosinófilos/patología , Eosinófilos/fisiología , Epitelio/metabolismo , Inmunización , Interleucina-5/genética , Pulmón/metabolismo , Pulmón/patología , Ratones , Ratones Noqueados , Ratones Transgénicos/genética , Ovalbúmina/inmunología , Eosinofilia Pulmonar/etiología , Eosinofilia Pulmonar/inmunología , Eosinofilia Pulmonar/patologíaRESUMEN
Overexpression of interleukin (IL)-5 by the airway epithelium in mice using the rat CC10 promoter (NJ.1726 line) leads to several histopathologies characteristic of human asthma, including airway hyperreactivity (AHR). We investigated the contribution of B and T cells, as well as CD4 expression, to the development of AHR in IL-5 transgenic mice. NJ.1726 mice on a T cell or CD4 knockout background, but not on a B cell knockout background, lost intrinsic AHR. These effects occurred without decreases in IL-5 or eosinophils. We further investigated the contribution of alpha(4)-integrin signaling to the development of AHR in IL-5 transgenic mice through the administration of anti-CD49d (alpha(4)-integrin) antibody (PS/2). Administration of PS/2 resulted in immediate (16-h) inhibition of AHR. The inhibition of AHR was not associated with a decrease in airway eosinophils. These studies demonstrate that, despite the presence of increased levels of IL-5 and eosinophils in the lungs of NJ.1726 mice, CD4(+) cells and alpha(4)-integrin signaling are necessary for the intrinsic AHR that develops in IL-5 transgenic mice.
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Antígenos CD/fisiología , Hiperreactividad Bronquial/etiología , Linfocitos T CD4-Positivos/fisiología , Interleucina-5/fisiología , Transducción de Señal/fisiología , Animales , Anticuerpos/farmacología , Antígenos CD/inmunología , Linfocitos B/citología , Bronquios/efectos de los fármacos , Bronquios/patología , Bronquios/fisiopatología , Hiperreactividad Bronquial/patología , Hiperreactividad Bronquial/fisiopatología , Líquido del Lavado Bronquioalveolar/química , Linfocitos T CD4-Positivos/patología , Eosinofilia/patología , Integrina alfa4 , Interleucina-5/genética , Pulmón/citología , Enfermedades Pulmonares/patología , Ratones , Ratones Transgénicos/genéticaRESUMEN
Paradigms of eosinophil effector function in the lungs of asthma patients invariably depend on activities mediated by cationic proteins released from secondary granules during a process collectively referred to as degranulation. In this study, we generated knockout mice deficient for eosinophil peroxidase (EPO) to assess the role(s) of this abundant secondary granule protein in an OVA-challenge model. The loss of EPO had no effect on the development of OVA-induced pathologies in the mouse. The absence of phenotypic consequences in these knockout animals extended beyond pulmonary histopathologies and airway changes, as EPO-deficient animals also displayed OVA-induced airway hyperresponsiveness after provocation with methacholine. In addition, EPO-mediated oxidative damage of proteins (e.g., bromination of tyrosine residues) recovered in bronchoalveolar lavage from OVA-treated wild-type mice was <10% of the levels observed in bronchoalveolar lavage recovered from asthma patients. These data demonstrate that EPO activities are inconsequential to the development of allergic pulmonary pathologies in the mouse and suggest that degranulation of eosinophils recruited to the lung in this model does not occur at levels comparable to those observed in humans with asthma.
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Eosinófilos/enzimología , Eosinófilos/inmunología , Pulmón/metabolismo , Pulmón/patología , Ovalbúmina/inmunología , Peroxidasas/metabolismo , Proteínas/metabolismo , Alérgenos/administración & dosificación , Alérgenos/inmunología , Animales , Hiperreactividad Bronquial/genética , Hiperreactividad Bronquial/inmunología , Hiperreactividad Bronquial/patología , Degranulación de la Célula/inmunología , Movimiento Celular/genética , Movimiento Celular/inmunología , Cruzamientos Genéticos , Gránulos Citoplasmáticos/metabolismo , Gránulos Citoplasmáticos/ultraestructura , Modelos Animales de Enfermedad , Peroxidasa del Eosinófilo , Eosinófilos/metabolismo , Eosinófilos/ultraestructura , Inyecciones Intraperitoneales , Pulmón/enzimología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ovalbúmina/administración & dosificación , Oxidación-Reducción , Peroxidasas/deficiencia , Peroxidasas/genética , Hipersensibilidad Respiratoria/enzimología , Hipersensibilidad Respiratoria/genética , Hipersensibilidad Respiratoria/patología , Eliminación de SecuenciaRESUMEN
Growing evidence suggests that intracellular vitamin D receptors are present in skeletal muscle tissue mediating vitamin D hormone response. The aim of the work reported here was to investigate the in situ expression of 1,25-dihydroxyvitamin D3 receptor in human skeletal muscle tissue. Intraoperative periarticular muscle biopsies were taken from 20 female orthopaedic patients (17 middle-aged and elderly patients receiving total hip arthroplasty due to osteoarthritis of the hip or an osteoporotic hip fracture and 3 young patients who received back surgery). The immunohistological distribution of the vitamin D3 receptor was investigated using a monoclonal rat antibody to the receptor (Clone Nr. 9A7). The receptor-positive nuclei were quantified by counting 500 nuclei per biopsy. Strong intranuclear immunostaining of the vitamin D receptor was detected in human muscle cells. Biopsies of hip patients had significantly fewer receptor-positive nuclei compared to those of back surgery patients (Mann-Whitney U-test: p = 0.0025). VDR expression (number of antigen-positive nuclei) was significantly correlated with age (coefficient of correlation = 0.46; p = 0.005), but not with 25-hydroxyvitamin D or 1,25-dihydroxyvitamin D levels. The data clearly demonstrate presence of nuclear 1,25-dihydroxyvitamin D3 receptor in human skeletal muscle. To our knowledge this is the first in situ detection of the receptor in human skeletal muscle. The difference in the expression of the receptor between hip and spinal muscle biopsies might be explained by age or location. Further research is needed in order to evaluate whether vitamin D3 receptor expression in human skeletal muscle is age-dependent and varies between different muscles.