RESUMO
Organ donors are sources of physiologically healthy organs and tissues for life-saving transplantation, and have been recently used for human immunology studies which are typically confined to the sampling of peripheral blood. Donors comprise a diverse population with different causes of death and clinical outcomes during hospitalization, and the effects of such variations on immune parameters in blood and tissues are not known. We present here a coordinate analysis of innate and adaptive immune components in blood, lymphoid (bone marrow, spleen, lymph nodes), and mucosal (lungs, intestines) sites from a population of brain-dead organ donors (2 months-93 years; n = 291) across eight clinical parameters. Overall, the blood of donors exhibited similar monocyte and lymphocyte content and low serum levels of pro-inflammatory cytokines as healthy controls; however, donor blood had increased neutrophils and serum levels of IL-8, IL-6, and MCP-1 which varied with cause of death. In tissues, the frequency and composition of monocytes, neutrophils, B lymphocytes and T cell subsets in lymphoid or mucosal sites did not vary with clinical state, and was similar in donors independent of the extent of clinical complications. Our results reveal that organ donors maintain tissue homeostasis, and are a valuable resource for fundamental studies in human immunology.
Assuntos
Morte Encefálica/imunologia , Linfócitos/imunologia , Células Mieloides/imunologia , Transplante de Órgãos , Doadores de Tecidos , Obtenção de Tecidos e Órgãos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Morte Encefálica/patologia , Estudos de Casos e Controles , Criança , Pré-Escolar , Citocinas/sangue , Feminino , Seguimentos , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Prognóstico , Subpopulações de Linfócitos T/imunologia , Adulto JovemRESUMO
OBJECTIVE: To assess bone-muscle (B-M) indices as risk factors for incident fractures in men. METHODS: Participants of the Osteoporotic Fractures in Men (MrOS) Study completed a peripheral quantitative computed tomography scan at 66% of their tibial length. Bone macrostructure, estimates of bone strength, and muscle area were computed. Areal bone mineral density (aBMD) and body composition were assessed with dual-energy X-ray absorptiometry. Four year incident non-spine and clinical vertebral fractures were ascertained. B-M indices were expressed as bone-to-muscle ratios for: strength, mass and area. Discriminative power and hazards ratios (HR) for fractures were reported. RESULTS: In 1163 men (age: 77.2±5.2 years, body mass index (BMI): 28.0±4.0 kg/m(2), 4.1±0.9 follow-up years, 7.7% of men â1 fracture), B-M indices were smaller in fractured men except for bending and areal indices. Smaller B-M indices were associated with increased fracture risk (HR: 1.30 to 1.74) independent of age and BMI. Strength and mass indices remained significant after accounting for lumbar spine but not total hip aBMD. However, aBMD correlated significantly with B-M indices. CONCLUSION: Mass and bending B-M indices are risk factors for fractures in men, but may not improve fracture risk prediction beyond that provided by total hip aBMD.
Assuntos
Osso e Ossos/patologia , Músculo Esquelético/patologia , Fraturas por Osteoporose/patologia , Absorciometria de Fóton , Idoso , Idoso de 80 Anos ou mais , Composição Corporal , Densidade Óssea , Força da Mão , Humanos , Masculino , Fraturas por Osteoporose/epidemiologia , Fatores de RiscoRESUMO
Congenital luxation of the ulnar and radius is a rare orthopaedic condition in the dog. This case report describes a novel surgical treatment for congenital elbow luxation in a medium-breed dog. A 6-week-old Kelpie presented for left forelimb lameness and deformity. Radiographs and computed tomography (CT) confirmed the diagnosis of unilateral ulnar and radius luxation. The surgical repair involved open reduction followed by fixation with an extraarticular pin and a transarticular external fixator. The outcome of surgery was improved leg function and weight-bearing, however, reluxation of the radial head was diagnosed 7 weeks following surgery. This technique may offer an alternative surgical option to improve clinical signs, as well as insight into improving the management of this condition.
Assuntos
Doenças do Cão , Luxações Articulares , Cães , Animais , Cotovelo , Luxações Articulares/diagnóstico por imagem , Luxações Articulares/cirurgia , Luxações Articulares/veterinária , Rádio (Anatomia)/cirurgia , Fixadores Externos/veterinária , Radiografia , Doenças do Cão/diagnóstico por imagem , Doenças do Cão/cirurgia , Doenças do Cão/congênito , Resultado do TratamentoRESUMO
The volume of bone erosions in the metacarpophalangeal joints is a radiological feature that can be used to track the progression of rheumatoid arthritis. We introduce a hybrid segmentation algorithm that combines region growing and level-set segmentation algorithms to semiautomatically measure the volume of bone erosions in magnetic resonance images. A total of 40 rheumatoid arthritis patients were included in the study. The scans of eight patients were used for training, whereas the remaining 32 scans were used to determine the accuracy, precision, and speed of the technique. The reproducibility of the semiautomated technique and that of manual segmentation was defined in terms of intraclass correlation coefficients. Both techniques were equally precise with intraclass correlation coefficient values greater than 0.9. The hybrid algorithm was highly accurate: the least squares fit between the semiautomated segmentations to those manually traced by a musculoskeletal radiologist resulted in a slope of 1.030 with an x-intercept of 1.385 mm(3) and an R(2) value of 0.923. The semiautomated technique was significantly faster than manual segmentation, which took two to four times longer to complete. Our hybrid algorithm shows promise in the quantitative assessment of radiological features of rheumatoid arthritis in a clinical setting.
Assuntos
Algoritmos , Artrite Reumatoide/patologia , Imageamento por Ressonância Magnética/métodos , Articulação Metacarpofalângica/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Imageamento Tridimensional , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Inquéritos e QuestionáriosRESUMO
INTRODUCTION: The sesamoid disease is a cause of lameness in dogs, and there is limited literature relating to diagnosis, treatment and outcome of treatment in dogs with the sesamoid disease. Our aim was to compare the efficacy of intra-articular metacarpophalangeal/metatarsophalangeal joint injection with methylprednisolone and bupivacaine (IMPB) or conservative management with nonsteroidal anti-inflammatories and rest (CMNR) for treatment of this disease. MATERIALS AND METHODS: We conducted a retrospective survey of dogs treated for the sesamoid disease with IMPB or CMNR. The medical records of all dogs that received IMPB or were recommended CMNR for treatment of sesamoid pain were reviewed, and a client questionnaire was delivered to owners. Response to treatment, rapidity of response, length of resolution and recurrence of clinical signs associated with the sesamoid disease were assessed. RESULTS: A total of 78 dogs were included in the study. One week after IMPB, 52/58 (89.7%) dogs demonstrated resolution of lameness compared with 1 week of CMNR, 0/18 (P < 0.001). There was limited statistical evidence in client satisfaction between treatment groups, IMPB 36/53 (67.9%) and CMNR 16/17 (94%) (P = 0.052). Dogs presenting with the sesamoid disease had comorbidities in 51/78 (65.4%) of cases. Elbow disease was the most common comorbidity 29/78 (37.2%). CONCLUSION: Our results support the use of IMPB for short-term (1 week) resolution of lameness associated with sesamoid disease in dogs. Dogs treated with CMNR had slower improvement; however, there was no difference in lameness or client satisfaction between treatment groups at long-term follow-up (12 months).
Assuntos
Bupivacaína , Doenças do Cão , Animais , Bupivacaína/uso terapêutico , Doenças do Cão/tratamento farmacológico , Cães , Injeções Intra-Articulares/veterinária , Coxeadura Animal/tratamento farmacológico , Coxeadura Animal/etiologia , Metilprednisolona/uso terapêutico , Estudos Retrospectivos , Resultado do TratamentoRESUMO
UNLABELLED: We examined the association of serum 25-hydroxyvitamin D [25(OH)D] with indices of bone quality in older men. Positive associations for 25(OH)D and bone mineral density, content, cortical thickness, and axial and polar strength strain indices were observed among Caucasians; however, among men of African descent findings were either null or negative. INTRODUCTION: There are limited data on serum 25(OH)D and bone measures in men of African ancestry. To better understand racial differences in vitamin D status and bone health, a cross-sectional study among 446 Caucasian men in the US and 496 men of African ancestry in Tobago (age ≥ 65 years) was conducted. METHODS: Serum 25(OH)D (liquid chromatography and tandem mass spectrometry) was measured, and peripheral quantitative computed tomography scans were administered. Bone measures estimated included trabecular and cortical volumetric bone mineral density (vBMD), bone mineral content (BMC), bone geometry (cross-sectional area and cortical thickness), and polar and axial strength strain indices (SSIp and SSIx). RESULTS: Men of African ancestry had higher 25(OH)D than Caucasians (34.7 vs. 27.6 ng/ml, p < 0.01). Among Caucasians, 25(OH)D was positively (p trend < 0.05) associated with cortical vBMD, total BMC, cortical thickness, SSIp, and SSIx at the distal radius after adjustment for potential confounders. Similar patterns were observed at the distal tibia. In contrast, in men of African ancestry, there was an inverse association (p trend < 0.05) between 25(OH)D and the cross-sectional area, and SSIx. Race modified (p for interaction < 0.05) the association between 25(OH)D and total BMC, cross-sectional area, SSIp, SSIx, and trabecular vBMD of the radius. In men of African ancestry, there was evidence of a threshold effect (at approximately 18 ng/ml) for 25(OH)D on tibial total BMC and cortical thickness. CONCLUSIONS: More studies are needed to better comprehend these race differences for 25(OH)D and bone density, geometry, and indices of bone strength.
Assuntos
Densidade Óssea/fisiologia , Rádio (Anatomia) , Tíbia , Vitamina D/análogos & derivados , Idoso , População Negra , Estudos Transversais , Humanos , Masculino , Pennsylvania , Rádio (Anatomia)/anatomia & histologia , Rádio (Anatomia)/fisiologia , Tíbia/anatomia & histologia , Tíbia/fisiologia , Trinidad e Tobago/etnologia , Vitamina D/sangue , População BrancaRESUMO
Intravascular large B cell lymphoma (IVLBCL) is a rare cause of pyrexia of unknown origin. Because of its protean clinical manifestations, diagnosis is elusive and is often made postmortem. We report here a case of IVLBCL that evaded diagnosis despite multiple investigations in vivo for pyrexia of unknown origin over a 5‐month period.
Assuntos
Febre de Causa Desconhecida/etiologia , Linfoma de Células B/complicações , Linfoma de Células B/diagnóstico , Neoplasias Vasculares/complicações , Neoplasias Vasculares/diagnóstico , Idoso , Antígenos CD20/análise , Evolução Fatal , Feminino , Humanos , Imuno-Histoquímica , Linfoma de Células B/patologia , Microscopia , Neoplasias Vasculares/patologiaRESUMO
BACKGROUND: . Severe pandemic 2009 influenza A virus (H1N1) infection is associated with risk factors that include pregnancy, obesity, and immunosuppression. After identification of immunoglobulin G(2) (IgG(2)) deficiency in 1 severe case, we assessed IgG subclass levels in a cohort of patients with H1N1 infection. METHODS: Patient features, including levels of serum IgG and IgG subclasses, were assessed in patients with acute severe H1N1 infection (defined as infection requiring respiratory support in an intensive care unit), patients with moderate H1N1 infection (defined as inpatients not hospitalized in an intensive care unit), and a random sample of healthy pregnant women. RESULTS: Among the 39 patients with H1N1 infection (19 with severe infection, 7 of whom were pregnant; 20 with moderate infection, 2 of whom were pregnant), hypoabuminemia (P < .001), anemia (P < .001), and low levels of total IgG (P= .01), IgG(1) (P= .022), and IgG(2) (15 of 19 vs 5 of 20; P= .001; mean value +/- standard deviation [SD], 1.8 +/- 1.7 g/L vs 3.4 +/- 1.4 g/L; P= .003) were all statistically significantly associated with severe H1N1 infection, but only hypoalbuminemia (P= .02) and low mean IgG(2) levels (P= .043) remained significant after multivariate analysis. Follow-up of 15 (79%) surviving IgG(2)-deficient patients at a mean (+/- SD) of 90 +/- 23 days (R, 38-126) after the initial acute specimen was obtained found that hypoalbuminemia had resolved in most cases, but 11 (73%) of 15 patients remained IgG(2) deficient. Among 17 healthy pregnant control subjects, mildly low IgG(1) and/or IgG(2) levels were noted in 10, but pregnant patients with H1N1 infection had significantly lower levels of IgG(2) (P= .001). CONCLUSIONS: Severe H1N1 infection is associated with IgG(2) deficiency, which appears to persist in a majority of patients. Pregnancy-related reductions in IgG(2) level may explain the increased severity of H1N1 infection in some but not all pregnant patients. The role of IgG(2) deficiency in the pathogenesis of H1N1 infection requires further investigation, because it may have therapeutic implications.
Assuntos
Deficiência de IgG/epidemiologia , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Influenza Humana/epidemiologia , Influenza Humana/virologia , Adolescente , Adulto , Idoso , Feminino , Humanos , Influenza Humana/patologia , Masculino , Pessoa de Meia-Idade , Gravidez , Adulto JovemRESUMO
Tissue-resident memory T (TRM) cells are increasingly associated with the outcomes of health and disease. TRM cells can mediate local immune protection against infections and cancer, which has led to interest in TRM cells as targets for vaccination and immunotherapies. However, these cells have also been implicated in mediating detrimental pro-inflammatory responses in autoimmune skin diseases such as psoriasis, alopecia areata, and vitiligo. Here, we summarize the biology of TRM cells established in animal models and in translational human studies. We review the beneficial effects of TRM cells in mediating protective responses against infection and cancer and the adverse role of TRM cells in driving pathology in autoimmunity. A further understanding of the breadth and mechanisms of TRM cell activity is essential for the safe design of strategies that manipulate TRM cells, such that protective responses can be enhanced without unwanted tissue damage, and pathogenic TRM cells can be eliminated without losing local immunity.
Assuntos
Doenças Autoimunes/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Doenças Transmissíveis/imunologia , Rejeição de Enxerto/imunologia , Memória Imunológica , Neoplasias/imunologia , Animais , Autoimunidade , Modelos Animais de Doenças , Humanos , CamundongosRESUMO
Temperature-sensitive mutations fall into two general classes: those generating thermolabile proteins; and those generating defects in protein synthesis, folding or assembly. Temperature-sensitive mutations at 17 sites in the gene for the coat protein of Phage P22 are of the latter class, preventing the productive folding of the polypeptide chain at restrictive temperature. We show here that, though the coat subunits interact intimately to form the viral shell, these temperature-sensitive folding (TSF) mutations were all recessive to wild type. The mutant polypeptide chains were not rescued by the presence of wild-type polypeptide chains. Missense substitutions in multimeric proteins frequently exhibit intragenic complementation; however, all pairs of coat protein TSF mutants tested failed to complement. The recessive phenotypes, absence of rescue and absence of intragenic complementation are all accounted for by the TSF defect, in which destabilization of a folding intermediate at restrictive temperature prevents the mutant chain from reaching the conformation required for subunit/subunit recognition. We suggest that absence of intragenic complementation should be a general property of TSF mutations in genes encoding multimeric proteins. The spectra of new loci identified by isolating second-site suppressors and synthetic lethals of temperature sensitive mutants will also differ depending on the nature of the defect. In the case of TSF mutations, where folding intermediates are defective rather than the native molecule, the spectra of other genes identified should shift from those whose products interact with the native molecule to those whose products influence the folding process.
Assuntos
Bacteriófago P22/genética , Proteínas do Capsídeo , Capsídeo/genética , Capsídeo/química , Genes Recessivos , Genes Supressores , Genes Virais , Teste de Complementação Genética , Mutagênese , Fenótipo , Desnaturação Proteica , Dobramento de Proteína , Estrutura Terciária de Proteína , Temperatura , Proteínas Estruturais Virais/genéticaRESUMO
We assessed the reproducibility of X-ray fluorescence-based lead measurements from multiple measurements made on a low-concentration plaster of paris phantom and in five subjects measured five times on two occasions. Over a 6-month period, 220 measurements of the same phantom were obtained and showed a standard deviation of 1.29 micrograms Pb (g plaster of paris)-1. The two sets of in vivo measurements were made 10 months apart and revealed a mean standard deviation of 3.4 micrograms Pb (g bone mineral)-1 and 5.1 micrograms Pb (g bone mineral)-1 for males and females, respectively. Our measured standard deviation exceeded by 20-30% the calculated standard deviation associated with a single measurement both in the phantom and in subjects. This indicates that some variance is introduced during the measurement process. Operator learning and consistency significantly minimized this increased variability. Measured lead concentrations of the left and right tibia in 14 subjects showed no significant differences between legs. As a result, either tibia can be sampled and compared over time. The levels of reproducibility we report here mean that X-ray fluorescence-based determinations of bone lead concentrations are reliable both over the short and long term. Thus, reasonably sized confidence intervals can be placed on detected changes in concentration and should permit acquisition of longitudinal data within a reasonable length of time.
Assuntos
Osso e Ossos/química , Radioisótopos de Cádmio , Chumbo/análise , Espectrometria por Raios X , Osso e Ossos/diagnóstico por imagem , Feminino , Humanos , Masculino , Modelos Estruturais , Radiografia , Reprodutibilidade dos TestesRESUMO
Bone lead levels for 367 active and 14 retired lead smelter workers were measured in vivo by X-ray fluorescence in May-June 1994. The bone sites of study were the tibia and calcaneus; magnitudes of concentration were used to gauge lead body burden. Whole blood lead readings from the workers generated a cumulative blood lead index (CBLI) that approximated the level of lead exposure over time. Blood lead values for 204 of the 381 workers were gathered from workers returning from a 10-month work interruption that ended in 1991; their blood level values were compared to their tibia and calcaneus lead levels. The resulting relations allowed constraints to be placed on the endogenous release of lead from bone in smelter works. Calcaneus lead levels were found to correlate strongly with those for tibia lead, and in a manner consistent with observations from other lead industry workers. Relations between bone lead concentration and CBLI demonstrated a distinctly nonlinear appearance. When the active population was divided by date of hire, a significant difference in the bone lead-CBLI slope emerged. After a correction to include the component of CBLI existing before the workers' employment at the smelter was made, this difference persisted. This implies that the transfer of lead from blood to bone in the workers has changed over time, possibly as a consequence of varying exposure conditions.
Assuntos
Osso e Ossos/química , Chumbo/análise , Exposição Ocupacional , Adulto , Idoso , Carga Corporal (Radioterapia) , Calcâneo/química , Canadá , Feminino , Humanos , Chumbo/sangue , Masculino , Metalurgia , Pessoa de Meia-Idade , Modelos Biológicos , Tíbia/químicaRESUMO
Hormone replacement therapy (HRT) in postmenopausal women suppresses the increase in bone resorption expected as circulating levels of endogenous estrogen decline. We tested the hypothesis that bone lead content might remain elevated in women on HRT. Fifty six women who at recruitment were on average 35 years postmenopausal were placed on calcium supplementation. Six months later 33 of these women were prescribed either low dose or moderate dose hormone replacement in addition to the calcium supplementation. After approximately 4 years of hormone replacement, lead content was measured at the tibia and calcaneus by in vivo fluorescence excitation, and lead concentrations were measured in serum, whole blood, and urine. Women not taking hormones had significantly lower lead concentrations in cortical bone compared to all women on HRT (p = 0.007). Tibia lead content (mean +/- SD) for women on calcium only was 11.13 +/- 6.22 microgram/g bone mineral. For women on HRT, tibia bone lead was 19.37 +/- 8.62 micrograms/g bone mineral on low-dose HRT and 16.87 +/- 11.68 micrograms/g bone mineral on moderate-dose HRT. There were no differences between groups for lead concentrations measured in trabecular bone, whole blood, serum or urine. Hormone replacement maintains cortical bone lead content. In women not on HRT, there will be a perimenopausal release of lead from bone.
Assuntos
Osso e Ossos/metabolismo , Terapia de Reposição de Estrogênios , Chumbo/metabolismo , Feminino , Humanos , Chumbo/sangue , Pessoa de Meia-IdadeRESUMO
Our purpose was to investigate the relationship between running volume and bone mineral mass in adult male runners. Whole body and regional bone mineral density were determined by dual-photon absorptiometry in 22 sedentary controls and 53 runners who were selected according to their running mileage to fall into a 5- to 10-, 15- to 20-, 25- to 30-, 40- to 55-, or 60- to 75-mile/wk group. All groups were of similar age (20-45 yr) and nutritional status, as determined by 7-day food records. Regional sites for bone density measurements included the trunk, spine, pelvis, thighs, and lower legs. In addition, serum total testosterone was determined in each subject and computed tomography scans were made of the lower legs in 34 subjects to assess bone cross-sectional area. No significant differences were detected for bone density measurements with the exception of the lower legs where it was significantly (P less than 0.05) greater for the 15- to 20-mile/wk group than for the control and 5- to 10-mile/wk groups. With mileage greater than 20 miles/wk, bone density of the lower legs showed no further increase and, in fact, tended to decrease, so that for the 60- to 75-mile/wk group it was similar to that of the controls. Cross-sectional area of the tibia and fibula when normalized to body weight tended to be greater as weekly mileage increased and was significantly greater in the 40- to 55-mile/wk runners than in the control group.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Densidade Óssea/fisiologia , Corrida/fisiologia , Testosterona/sangue , Adulto , Fíbula/anatomia & histologia , Fíbula/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Tíbia/anatomia & histologia , Tíbia/metabolismo , Distribuição TecidualRESUMO
In this study a method of assessing trabecular bone structure at the distal end of the radius from high-resolution magnetic resonance images is described. Trabecular bone is segmented from the marrow and soft tissue background using an adaptive threshold, a region growth, and a skeletonization step. From the processed image we measured the connectivity and orientation of the trabecular bone network. Connectivity was assessed by a proposed connectivity index (CI) and marrow space was quantitated by a mean hole area (HA). Significant age-related changes in CI and HA were observed in a mixed group of normal volunteers. CI decreased at a rate of 0.18 yr-1 (r = 0.72, n = 14, p < 0.05) and HA increased at a rate of 0.018 mm2 yr-1 (r = 0.69, n = 14, p < 0.05). Gradient analysis was used to examine trabecular orientation, and revealed that the individual trabeculae at the distal end of the radius are organized anisotropically along the bone. These findings suggest that clinical magnetic resonance scanners can be used to assess trabecular bone structure in vivo.
Assuntos
Desenvolvimento Ósseo , Osso e Ossos/anatomia & histologia , Imageamento por Ressonância Magnética/métodos , Imagens de Fantasmas , Adolescente , Adulto , Idoso , Envelhecimento , Medula Óssea/anatomia & histologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Rádio (Anatomia)/anatomia & histologia , Reprodutibilidade dos TestesRESUMO
Radiographic joint-space narrowing (JSN) is the principle indicator of cartilage loss in osteoarthritis (OA). JSN is usually assessed qualitatively by visual inspection or in clinical research, is measured manually with a graduated handheld lens directly applied to the x-ray film, or from digitized radiographs by hand tracing the joint margins with a mouse. The minimum joint-space width (mJSW) and joint-space area (JSA) are recorded as the indices of OA progression in epidemiological studies and clinical drug trials. We present a computerized method that automatically finds the articular margins of the hip to improve determination of mJSW and JSA. The algorithm requires that three seed points are manually identified on the femoral head and uses three steps to process each digitized hip x-ray. First, a Hough transform finds the center and radius (R) of a circle that approximates the femoral head. Finding R indicates whether magnification differences must be corrected on repeat exams. Second, a gradient algorithm finds the edge of the femoral head and acetabulum. Third, the mid-line of the femoral neck is automatically found and used to define the joint portion (theta) that is assessed for narrowing. theta is fixed for follow-up exams of the same subject. The algorithm was evaluated in three ways to determine its performance characteristics. First, the inter-reader and intra-reader variability for mJSW and JSA associated with the selection of the seed points was found to be negligible (< 1%) compared to the variability associated with manual scoring with a lens or by tracing the joint margins with a mouse. Second, from duplicate hip x-rays of 19 subjects with OA, the Root Mean Square Standard Deviation and coefficient of variation for mJSW and JSA defined by the algorithm was determined to be better than manual techniques by at least a factor of 2. Third, the algorithm correctly identified the joint margin in more than 85% of the 105 cases tested. Automated measures of radiographic hip joint-space narrowing is less subjective than manual methods and may be applicable for monitoring OA progression in clinical research.
Assuntos
Diagnóstico por Computador/métodos , Articulação do Quadril/diagnóstico por imagem , Osteoartrite/diagnóstico por imagem , Acetábulo/diagnóstico por imagem , Algoritmos , Fenômenos Biofísicos , Biofísica , Diagnóstico por Computador/estatística & dados numéricos , Cabeça do Fêmur/diagnóstico por imagem , Colo do Fêmur/diagnóstico por imagem , Humanos , Intensificação de Imagem Radiográfica , Reprodutibilidade dos TestesRESUMO
Although Aspergillus niger is used as a host for heterologous protein production, yields are generally lower than those obtained for homologous proteins. Mechanisms of protein secretion and the secretory pathway in filamentous fungi are poorly characterised, although there is evidence to suggest that secretion occurs by a mechanism similar to that in other eukaryotes, but with proteins destined for secretion being directed to the hyphal tip. We report on a method using a glucoamylase: GFP gene fusion which allows us for the first time to monitor, in vivo, protein secretion in A. niger at the single hyphal level. A synthetic green fluorescent protein (sGFP(S65T)) was fused to truncated A. niger glucoamylase (GLA:499). Southern blot analysis of transformants confirmed that the gene fusion had successfully integrated into the A. niger genome. Confocal and fluorescence microscopy revealed that the GLA::GFP fusion protein is fluorescent in A. niger and appears to be directed to the hyphal tip. In young mycelia, hyphal cell wall fluorescence is apparent and immunogold labelling of GFP confirmed that GFP was partially localised within the hyphal cell wall. Using Western blotting, extracellular GLA::GFP was detected only in culture filtrates of young mycelia grown in a soya milk medium. The actin inhibitor latrunculin B was used to disrupt the secretion process, and its effects on the distribution of GLA::GFP were monitored.
Assuntos
Aspergillus niger/genética , Aspergillus niger/metabolismo , Glucana 1,4-alfa-Glucosidase/genética , Glucana 1,4-alfa-Glucosidase/metabolismo , Proteínas Luminescentes/genética , Actinas/metabolismo , Aspergillus niger/crescimento & desenvolvimento , Southern Blotting , Western Blotting , Compostos Bicíclicos Heterocíclicos com Pontes/metabolismo , Meios de Cultura , Proteínas de Fluorescência Verde , Proteínas Luminescentes/metabolismo , Microscopia de Fluorescência , Plasmídeos/genética , Proteínas Recombinantes de Fusão/metabolismo , Tiazóis/metabolismo , TiazolidinasRESUMO
A dedicated computed tomography system was used to acquire transaxial images of the distal radius to assess trabecular bone structure in vivo. We segmented trabecular bone from the marrow and soft tissue background by postprocessing the image with a region grow and skeletonization step. From the processed image we assessed the integrity of the bone by examining the continuity of its trabecular network and by determining the area of the holes comprising its marrow space. The continuity of the bone imaged was assessed by a proposed connectivity index (CI) and the size of the marrow spaces was assessed by calculating a mean hole area (H(A)) in the bone cross-section. Repeat measurements revealed that the intra-subject variability in CI and H(A) was small (CV < 6%). Both CI and H(A) were sensitive enough to reflect differences in structure at the head of the radius and at several sites along its shaft. We tested the diagnostic value of assessing bone structure at the distal end of the radius by measuring trabecular bone density, CI and H(A) in a mixed group of 26 subjects, nine of whom had suffered a wrist fracture. We found that a trabecular bone density threshold of 116 mg cm-3, corresponding to two standard deviations below the mean density in the 17 non-fractured subjects, separated fractured from non-fractured subjects with a sensitivity of 22% and a specificity of 100%. A CI threshold of -4.7 doubled the sensitivity (44%) and maintained the 100% specificity. An H(A) threshold of 4.5 mm2 achieved a sensitivity of 89% and a specificity of 94%. This increased sensitivity achieved by our indices suggests that an in vivo assessment of trabecular bone structure can contribute significantly to the identification of persons at risk of fracture.
Assuntos
Osso e Ossos/diagnóstico por imagem , Tomografia Computadorizada por Raios X/métodos , Adulto , Idoso , Algoritmos , Fenômenos Biofísicos , Biofísica , Densidade Óssea , Feminino , Fraturas Ósseas/diagnóstico por imagem , Humanos , Pessoa de Meia-Idade , Interpretação de Imagem Radiográfica Assistida por Computador , Rádio (Anatomia)/diagnóstico por imagem , Fraturas do Rádio/diagnóstico por imagem , Reprodutibilidade dos Testes , Tomografia Computadorizada por Raios X/estatística & dados numéricosRESUMO
In order to assess the accuracy of peripheral QCT (Stratec XCT 960) we analysed scans of the European Forearm Phantom and another phantom consisting of K2HPO4 encased in aluminium tubes to simulate cortical walls. Additionally 14 cadaveric forearm specimen scans were compared to CT scans acquired on a GE9800Q. The accuracy for density assessment of the European Forearm Phantom was better than 3%. A small increase in density was observed with increasing thickness of the aluminium wall (10% for each mm). Density measurements within the wall were confounded by limited spatial resolution. For a thickness of less than 4 mm, the density within the wall was underestimated by up to 40%. The measurement of mineral content was not influenced by this effect and showed an accuracy error of less than 6%. The agreement of density measurements on the different CT systems was very strong (R2 > 0.96; RMSE < 6.2%). Our findings suggest that the Stratec pQCT scanner very accurately measures volumetric trabecular and total bone mineral densities at the distal radius while the assessment of cortical density is associated with considerable inaccuracies due to limited spatial resolution.
Assuntos
Osso e Ossos/diagnóstico por imagem , Antebraço/diagnóstico por imagem , Tomografia Computadorizada por Raios X/métodos , Alumínio , Densidade Óssea , Cadáver , Estudos de Avaliação como Assunto , Fraturas Ósseas/diagnóstico , Humanos , Fosfatos/química , Tomógrafos Computadorizados , Tomografia Computadorizada por Raios X/instrumentaçãoRESUMO
In 111 subjects not occupationally exposed, bone lead content increased steadily with age in both men and women. Higher than expected bone lead levels were observed in two-thirds of 27 subjects working in occupations with potential for lead exposure. Five of 8 patients who displayed symptoms which might have been due to lead poisoning had increased bone lead levels. In vivo bone lead measurements reflect the cumulative extent of exposure to environmental and occupational sources of lead and allow the assessment of abnormal exposures.