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1.
Theor Appl Genet ; 136(4): 72, 2023 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-36952017

RESUMO

KEY MESSAGE: Here, we provide an updated set of guidelines for naming genes in wheat that has been endorsed by the wheat research community. The last decade has seen a proliferation in genomic resources for wheat, including reference- and pan-genome assemblies with gene annotations, which provide new opportunities to detect, characterise, and describe genes that influence traits of interest. The expansion of genetic information has supported growth of the wheat research community and catalysed strong interest in the genes that control agronomically important traits, such as yield, pathogen resistance, grain quality, and abiotic stress tolerance. To accommodate these developments, we present an updated set of guidelines for gene nomenclature in wheat. These guidelines can be used to describe loci identified based on morphological or phenotypic features or to name genes based on sequence information, such as similarity to genes characterised in other species or the biochemical properties of the encoded protein. The updated guidelines provide a flexible system that is not overly prescriptive but provides structure and a common framework for naming genes in wheat, which may be extended to related cereal species. We propose these guidelines be used henceforth by the wheat research community to facilitate integration of data from independent studies and allow broader and more efficient use of text and data mining approaches, which will ultimately help further accelerate wheat research and breeding.


Assuntos
Melhoramento Vegetal , Triticum , Triticum/genética , Fenótipo , Genes de Plantas , Grão Comestível/genética
2.
Diabet Med ; 35(6): 760-769, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29478265

RESUMO

AIMS: To assess the prevalence and management of depressive disorders in people with Type 2 diabetes in different countries. METHODS: People with diabetes aged 18-65 years and treated in outpatient settings were recruited in 14 countries and underwent a psychiatric interview. Participants completed the Patient Health Questionnaire and the Problem Areas in Diabetes scale. Demographic and medical record data were collected. RESULTS: A total of 2783 people with Type 2 diabetes (45.3% men, mean duration of diabetes 8.8 years) participated. Overall, 10.6% were diagnosed with current major depressive disorder and 17.0% reported moderate to severe levels of depressive symptomatology (Patient Health Questionnaire scores >9). Multivariable analyses showed that, after controlling for country, current major depressive disorder was significantly associated with gender (women) (P<0.0001), a lower level of education (P<0.05), doing less exercise (P<0.01), higher levels of diabetes distress (P<0.0001) and a previous diagnosis of major depressive disorder (P<0.0001). The proportion of those with either current major depressive disorder or moderate to severe levels of depressive symptomatology who had a diagnosis or any treatment for their depression recorded in their medical records was extremely low and non-existent in many countries (0-29.6%). CONCLUSIONS: Our international study, the largest of this type ever undertaken, shows that people with diabetes frequently have depressive disorders and also significant levels of depressive symptoms. Our findings indicate that the identification and appropriate care for psychological and psychiatric problems is not the norm and suggest a lack of the comprehensive approach to diabetes management that is needed to improve clinical outcomes.


Assuntos
Transtorno Depressivo Maior/epidemiologia , Diabetes Mellitus Tipo 2/psicologia , Adolescente , Adulto , Idoso , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/epidemiologia , Feminino , Saúde Global , Humanos , Hipoglicemiantes/uso terapêutico , Insulina/uso terapêutico , Masculino , Pessoa de Meia-Idade , Prevalência , Adulto Jovem
3.
Opt Express ; 24(8): 8360-9, 2016 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-27137273

RESUMO

Ptychography is a scanning coherent diffractive imaging (CDI) technique that relies upon a high level of stability of the illumination during the course of an experiment. This is particularly an issue for coherent short wavelength sources, where the beam intensity is usually tightly focused on the sample in order to maximize the photon flux density on the illuminated region of the sample and thus a small change in the beam position results in a significant change in illumination of the sample. We present an improved ptychographic method that allows for limited stability of the illumination wavefront and thus significantly improve the reconstruction quality without additional prior knowledge. We have tested our reconstruction method in a proof of concept experiment, where the beam instability of a visible light source was emulated using a piezo driven mirror, and also in a short wavelength microscopy CDI setup using a high harmonic generation source in the extreme ultraviolet range. Our work shows a natural extension of the ptychography method that paves the way to use ptychographic imaging with any limited pointing stability coherent source such as free electron or soft X-ray lasers and improve reconstruction quality of long duration synchrotron experiments.

4.
J Bone Joint Surg Am ; 106(9): 840-844, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38252737

RESUMO

ABSTRACT: Most health systems are vertically integrated, and the leaders of orthopaedic surgery departments or service lines must have a comprehensive understanding of their role in the strategic plan of the health system. Orthopaedic surgery departments must be profitable while supporting the tripartite mission of excellence in clinical care, research, and education. This symposium had 4 specific objectives: to discuss how to (1) create synergy between the department or service line and the health system, (2) develop a strategy to enhance financial stability and revenue growth, (3) develop a comprehensive plan to enhance recruitment and retention of a diverse faculty, and (4) consider alternative strategies to foster education and research, even when the health system may be more focused on revenue generation.


Assuntos
Liderança , Ortopedia , Ortopedia/organização & administração , Humanos , Estados Unidos
5.
Appl Radiat Isot ; 194: 110680, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36696752

RESUMO

Recycling and conventional reuse of lead materials and structures originating from controlled areas in nuclear facilities rely on historical knowledge and well selected characterization procedures. At the SCK•CEN, one of these procedures involves performing high-resolution gamma-ray spectrometry measurements on several cylindrical shaped test samples (50 mm diameter and approximately 5 mm thickness), obtained during the lead melting campaigns. The high density (11.3 g/cm³) of these samples is a challenge for radionuclide analysis by gamma-ray spectrometry since no such calibration sources nor reference materials are available. We used the efficiency transfer procedure, relying on regular standard sources available in our laboratory, to set up calibrations for this specific counting geometry. The method proves to be fit for purpose.

6.
Appl Radiat Isot ; 193: 110646, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36603458

RESUMO

This work reports a new approach for the determination of 36Cl in radioactive waste samples from nuclear decommissioning, wherein novel plastic scintillator (PS) materials were used for the concentration of 36Cl prior to the detection with scintillation counting. Different plastic scintillator (PS) materials were tested for their selective absorption and detection of 36Cl activity in solid samples. PS microspheres (PSm), cross-linked PSm (CPSm) and PS resin have been investigated. PS resin was identified as the most suitable material for 36Cl analysis. Pyrolysis and subsequent trapping of the volatile elements in a bubbler was used. The trapping solution was finally loaded onto a cartridge of the PS resin. Scintillation counting and ion chromatography were used to determine the activity concentration and the chemical recovery, respectively.

7.
Appl Radiat Isot ; 202: 111046, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37778141

RESUMO

This study compares different approaches for the quantification of the massic activity of 36Cl in graphite samples. All approaches consisted of a combustion step in combination with a trapping solution to collect the volatile elements. Two different resins were used to separate 36Cl from the matrix (CL resin and PS resin). Liquid scintillation counting (LSC), scintillation counting (SC) and tandem inductively coupled plasma mass spectrometry (ICP-MS/MS) were used to quantify 36Cl activity. The chemical yield in all approaches was determined by means of ion chromatography (IC). In addition, the methods were applied to a real activated graphite sample.

8.
Nanotechnology ; 23(39): 395302, 2012 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-22972003

RESUMO

We have fabricated and measured single domain wall magnetoresistance devices with sub-20 nm gap widths using a novel combination of electron beam lithography and helium ion beam milling. The measurement wires and external profile of the spin valve are fabricated by electron beam lithography and lift-off. The critical bridge structure is created using helium ion beam milling, enabling the formation of a thinner gap (and so a narrower domain wall) than that which is possible with electron beam techniques alone. Four-point probe resistance measurements and scanning electron microscopy are used to characterize the milled structures and optimize the He ion dose. Successful operation of the device as a spin valve is demonstrated, with a 0.2% resistance change as the external magnetic field is cycled. The helium ion beam milling efficiency as extracted from electrical resistance measurements is 0.044 atoms/ion, about half the theoretical value. The gap in the device is limited to a maximum of 20 nm with this technique due to sub-surface swelling caused by injected ions which can induce catastrophic failure in the device. The fine patterning capabilities of the helium ion microscope milling technique indicate that sub-5 nm constriction widths could be possible.

9.
Appl Radiat Isot ; 134: 194-199, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28802626

RESUMO

This paper provides guidance for determining the neutron activation profile of core drill samples taken from the biological shield of nuclear reactors using gamma spectrometry measurements. Thus, it provides guidance for selecting a model of the right form to fit data and using least squares methods for model fitting. The activity profiles of two core samples taken from the biological shield of a nuclear reactor were determined. The effective activation depth and the total activity of core samples along with their uncertainties were computed by Monte Carlo simulation.

10.
Int J Biochem Cell Biol ; 32(10): 1069-74, 2000 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-11091139

RESUMO

In order to elucidate the role of mitogen-activated protein kinase kinase (MEK-1/2) in 5-lipoxygenase (5-LO) activation we studied the N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced 5-LO translocation in human blood neutrophils (PMNs). In non-primed, Ca(2+)-repleted PMNs, fMLP consistently stimulated MEK-1/2 phosphorylation, but induced 5-LO translocation and product formation (430+/-128 pmol; SEM, n=13) only in 13 of 18 PMN preparations from different healthy donors. In fMLP-responsive cells, the MEK-1/2 inhibitor PD098059 (50 microM) attenuated MEK phosphorylation and abolished 5-LO activation at the translocation step. The fMLP-mediated 5-LO product formation was also sensitive to MEK inhibition by U0126 and to p38 inhibition by SB203580. But in contrast to PD098059, U0126 at 10 microM and SB203580 at 20-50 microM impaired 5-LO activity in the cell-free assay setting, suggesting direct actions of higher concentrations of U0126 and SB203580 on 5-LO apart from MEK and p38 inhibition, respectively. These data show that fMLP initiates 5-LO product formation in non-primed, Ca(2+)-repleted human blood PMNs from healthy donors, and that MEK signaling is pivotal, but not sufficient for 5-LO activation in response to the receptor agonist fMLP.


Assuntos
Araquidonato 5-Lipoxigenase/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Quinases de Proteína Quinase Ativadas por Mitógeno/antagonistas & inibidores , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Neutrófilos/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/antagonistas & inibidores , Butadienos/farmacologia , Calcimicina/farmacologia , Cálcio/metabolismo , Cálcio/farmacologia , Sistema Livre de Células , Ativação Enzimática/efeitos dos fármacos , Flavonoides/farmacologia , Humanos , Imidazóis/farmacologia , MAP Quinase Quinase 1 , MAP Quinase Quinase 2 , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Neutrófilos/enzimologia , Neutrófilos/metabolismo , Nitrilas/farmacologia , Fosforilação/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/metabolismo , Transporte Proteico/efeitos dos fármacos , Proteínas Tirosina Quinases/metabolismo , Piridinas/farmacologia
11.
Endocrinology ; 134(3): 1429-36, 1994 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-8119183

RESUMO

Osteoblasts have been reported to produce tissue-type (t) plasminogen activator (PA), which may be involved in the initiation of bone resorption via plasmin-metalloproteinase degradation of adjacent extracellular matrix. To investigate this cAMP-activated gene, we characterized the PTH regulation of tPA messenger RNA (mRNA) in neonatal rat osteoblast cultures before and after differentiation in vitro. RNA was purified from cultures at confluence or after treatment with glucocorticoid for 1 week and BGJ/ascorbic acid/beta-glycerophosphate for a second week. Northern blots of total or poly(A)+ RNA were hybridized simultaneously with an oligonucleotide or complementary RNA probe for rat tPA and an oligomeric DNA probe for cyclophilin (CYP), an abundant control gene. Differentiation was monitored by expression of rat osteocalcin mRNA and protein. Both bovine PTH1-34 and forskolin caused an increase in tPA/CYP ratio in the presence of phosphodiesterase inhibitor (IBMX) and cycloheximide (CHX). The effect was maximal (16- to 21-fold increase in tPA mRNA and 6- to 8-fold increase in tPA/CYP ratio) with 25 nM hormone for 6 h and was half-maximally stimulated by 0.75-2.5 nM PTH. The tPA response to PTH was present in first passage osteoblast cultures at confluence and after 1 to 2 weeks of glucocorticoid treatment. Exposure of the differentiated cultures of 1,25-dihydroxyvitamin D (10 nM) for 2 days markedly stimulated osteocalcin mRNA while having no effect on tPA. In Northern blots of poly(A)+ RNA from cultures not treated with CHX, IBMX and PTH (2.5 h) independently stimulated tPA mRNA with no significant effect on CYP mRNA levels. The tPA/CYP ratio increased in five consecutive experiments and the effect of IBMX and PTH were additive. These data indicate that PTH acts via cAMP to stimulate tPA expression by a mechanism that is independent of protein synthesis. The enhancement of PTH action by CHX is compatible with feedback inhibition of tPA transcription by a hormone-activated repressor (which has been proposed to occur in granulosa cells) but effects of CHX on tPA mRNA stability may also occur. Expression of tPA mRNA before and after differentiation may indicate that the enzyme has more than one function.


Assuntos
Osteoblastos/efeitos dos fármacos , Hormônio Paratireóideo/farmacologia , RNA Mensageiro/análise , Ativador de Plasminogênio Tecidual/genética , Animais , Animais Recém-Nascidos , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , AMP Cíclico/fisiologia , Cicloeximida/farmacologia , Glucocorticoides/farmacologia , Osteoblastos/metabolismo , Ratos
12.
Endocrinology ; 141(11): 3956-64, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11089525

RESUMO

Tumor necrosis factor-alpha (TNF-alpha) has a key role in skeletal disease in which it promotes reduced bone formation by mature osteoblasts and increased osteoclastic resorption. Here we show that TNF inhibits differentiation of osteoblasts from precursor cells. TNF-alpha treatment of fetal calvaria precursor cells, which spontaneously differentiate to the osteoblast phenotype over 21 days, inhibited differentiation as shown by reduced formation of multilayered, mineralizing nodules and decreased secretion of the skeletal-specific matrix protein osteocalcin. The effect of TNF was dose dependent with an IC50 of 0.6 ng/ml, indicating a high sensitivity of these precursor cells. Addition of TNF-alpha from days 2-21, 2-14, 7-14, and 7-10 inhibited nodule formation but addition of TNF after day 14 had no effect. Partial inhibition of differentiation was observed with addition of TNF on only days 7-8, suggesting that TNF could act during a critical period of phenotype selection. Growth of cells on collagen-coated plates did not prevent TNF inhibition of differentiation, suggesting that inhibition of collagen deposition into matrix by proliferating cells could not, alone, explain the effect of TNF. Northern analysis revealed that TNF inhibited the expression of insulin-like growth factor I (IGF-I). TNF had no effect on expression of the osteogenic bone morphogenic proteins (BMPs-2, -4, and -6), or skeletal LIM protein (LMP-1), as determined by semiquantitative RT-PCR. Addition of IGF-I or BMP-6 to fetal calvaria precursor cell cultures enhanced differentiation but could not overcome TNF inhibition, suggesting that TNF acted downstream of these proteins in the differentiation pathway. The clonal osteoblastic cell line, MC3T3-E1-14, which acquires the osteoblast phenotype spontaneously in postconfluent culture, was also studied. TNF inhibited differentiation of MC3T3-E1-14 cells as shown by failure of mineralized matrix formation in the presence of calcium and phosphate. TNF was not cytotoxic to either cell type as shown by continued attachment and metabolism in culture, trypan blue exclusion, and Alamar Blue cytotoxicity assay. These results demonstrate that TNF-alpha is a potent inhibitor of osteoblast differentiation and suggest that TNF acts distal to IGF-I, BMPs, and LMP-1 in the progression toward the osteoblast phenotype.


Assuntos
Diferenciação Celular , Osteoblastos/citologia , Fator de Necrose Tumoral alfa/farmacologia , Proteínas Adaptadoras de Transdução de Sinal , Animais , Ácido Ascórbico/farmacologia , Northern Blotting , Proteínas Morfogenéticas Ósseas/genética , Osso e Ossos/embriologia , Fosfatos de Cálcio/metabolismo , Proteínas de Transporte/genética , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Proteínas do Citoesqueleto , Expressão Gênica , Fator de Crescimento Insulin-Like I/genética , Peptídeos e Proteínas de Sinalização Intracelular , Proteínas com Domínio LIM , Fosfatos/administração & dosagem , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa
13.
Endocrinology ; 138(7): 2820-8, 1997 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-9202223

RESUMO

Glucocorticoids (GCs) at physiological concentrations promote osteoblast differentiation from fetal calvarial cells, calvarial organ cultures, and bone marrow stromal cells; however, the cellular pathways involved are not known. Bone morphogenetic proteins (BMPs) are recognized as important mediators of osteoblast differentiation. Specific roles for individual BMPs during postembryonic membranous bone formation have yet to be determined. We recently reported that GC potentiated the osteoblast differentiation effects of BMP-2 and BMP-4, but not of BMP-6, which, by itself, was the most potent of the three. In the present study, we used fetal rat secondary calvarial cultures to study the role of BMP-6 during early osteoblast differentiation. Treatment with the GC triamcinolone (10(-9) M) resulted in a 5- to 8-fold increase in BMP-6 steady-state messenger RNA levels, peaking at 12 h. In contrast, BMPs -2, -4, -5, -7, and transforming growth factor (TGF)-beta1 messenger RNA levels increased by less than 2-fold, after GC treatment, compared with untreated control cultures at 24 h. BMP-6 protein secretion increased 6- to 7-fold by 12 h and 12-fold (from 7.5 to 90 ng/ml) by 24 h, as measured by quantitative Western analysis. Treatment of cells with oligodeoxynucleotides antisense to BMP-6 diminished secretion of BMP-6 protein and significantly inhibited the GC-induced differentiation, as determined by a 10-fold decrease in the number of mineralized bone nodules, compared with controls that were treated with sense oligonucleotides or no oligonucleotides (ANOVA, P < 0.05). The antisense oligonucleotide inhibition of differentiation was rescued by treatment with exogenous recombinant human BMP-6. We conclude that GC-induced differentiation of osteoblasts from the pluripotent precursors is mediated, in part, by BMP-6. These results suggest that BMP-6 has an important and unique role during early osteoblast differentiation.


Assuntos
Proteínas Morfogenéticas Ósseas/metabolismo , Glucocorticoides/farmacologia , Osteoblastos/citologia , Crânio/embriologia , Animais , Western Blotting , Proteína Morfogenética Óssea 6 , Proteínas Morfogenéticas Ósseas/genética , Células CHO , Diferenciação Celular/efeitos dos fármacos , Cricetinae , Humanos , Oligonucleotídeos Antissenso/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes/metabolismo , Fatores de Tempo
14.
Endocrinology ; 139(12): 5125-34, 1998 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9832452

RESUMO

Glucocorticoids can promote osteoblast differentiation from fetal calvarial cells and bone marrow stromal cells. We recently reported that glucocorticoid specifically induced bone morphogenetic protein-6 (BMP-6), a glycoprotein signaling molecule that is a multifunctional regulator of vertebrate development. In the present study, we used fetal rat secondary calvarial cultures to determine genes induced during early osteoblast differentiation as initiated by glucocorticoid treatment. Glucocorticoid, and subsequently BMP-6, was found to induce a novel rat intracellular protein, LIM mineralization protein-1 (LMP-1), that in turn resulted in synthesis of one or more soluble factors that could induce de novo bone formation. Blocking expression of LMP-1 using antisense oligonucleotide prevented osteoblast differentiation in vitro. Overexpression of LMP-1 using a mammalian expression vector was sufficient to initiate de novo bone nodule formation in vitro and in sc implants in vivo. These data demonstrate that LMP-1 is an essential positive regulator of the osteoblast differentiation program as well as an important intermediate step in the BMP-6 signaling pathway.


Assuntos
Proteínas Morfogenéticas Ósseas/fisiologia , Proteínas de Transporte/fisiologia , Osteogênese/fisiologia , Proteínas Adaptadoras de Transdução de Sinal , Sequência de Aminoácidos , Animais , Células da Medula Óssea/fisiologia , Proteína Morfogenética Óssea 6 , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Diferenciação Celular/fisiologia , Proteínas do Citoesqueleto , DNA Complementar/genética , Regulação da Expressão Gênica/fisiologia , Glucocorticoides/farmacologia , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Proteínas com Domínio LIM , Dados de Sequência Molecular , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley
15.
Endocrinology ; 137(8): 3401-7, 1996 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-8754767

RESUMO

Bone morphogenetic proteins (BMPs) induce cartilage and bone differentiation in vivo and promote osteoblast differentiation from calvarial and marrow stromal cell preparations. Functional differences between BMP-2, -4, and -6 are not well understood. Recent investigations find that these three closely related osteoinductive proteins may exert different effects in primary rat calvarial cell cultures, suggesting the possibility of unique functions in vivo. In this study, we use a fetal rat secondary calvarial cell culture system to examine the differential effects of BMP-2, -4, and -6 on early osteoblast differentiation. These cells do not spontaneously differentiate into osteoblasts, as do cells in primary calvarial cultures, but rather require exposure to a differentiation initiator such as glucocorticoid or BMP. We determined that BMP-6 is a 2- to 2.5-fold more potent inducer of osteoblast differentiation than BMP-2 or -4. BMP-6 induced the formation of more and larger bone nodules as well as increased osteocalcin secretion. The effects of all three of these BMPs were potentiated up to 10-fold by cotreatment or pretreatment with the glucocorticoid triamcinolone (Trm). The Trm effects were synergistic with those of BMP-2 or -4, suggesting that this glucocorticoid may increase the cell responsiveness to these BMPs. Finally, BMP-6 did not require either cotreatment or pretreatment with Trm to achieve greater amounts of osteoblast differentiation than seen with BMP-2 or BMP-4 treatment, suggesting that BMP-6 may act at an earlier stage of cell differentiation.


Assuntos
Proteínas Morfogenéticas Ósseas/farmacologia , Osteoblastos/citologia , Triancinolona Acetonida/farmacologia , Animais , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Sinergismo Farmacológico , Substâncias de Crescimento/farmacologia , Osteoblastos/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley
16.
Tissue Eng ; 6(4): 383-99, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10992434

RESUMO

Posterolateral lumbar spine arthrodesis is a commonly performed procedure, yet the biology of healing is poorly understood. Nonunion, or failure to achieve a solid bony fusion, occurs in up to 40% of patients. We first developed and validated a rabbit model to characterize the healing process by measuring macroscopic parameters, microscopic parameters, and gene expression. We found that presently available osteoconductive and weakly osteoinductive materials were insufficient to replace autografts, but could in some cases serve as bone graft extenders. In contrast, two osteoinductive growth factors currently in development could replace autograft in non-human primates and in humans, but may be limited by the high dose required, carrier variability, and high cost. We identified, cloned, and sequenced a novel complementary DNA (cDNA) encoding for an intracellular protein LMP-1, which is expressed during the first few hours of osteoblast differentiation. LMP-1 expression is able to induce many BMPs, their receptors, and other bone growth factors. Local implantation of bone marrow cells transfected with LMP-1 cDNA induced spine fusion in 100% of sites tested; no bone formed at the control sites without LMP-1. This strategy of local gene therapy may provide a basis for the next generation of bone graft substitutes.


Assuntos
Substitutos Ósseos , Vértebras Lombares , Doenças da Coluna Vertebral/terapia , Fusão Vertebral/métodos , Proteínas Adaptadoras de Transdução de Sinal , Animais , Transplante de Medula Óssea , Proteínas Morfogenéticas Ósseas/genética , Proteínas Morfogenéticas Ósseas/fisiologia , Proteínas de Transporte/genética , Proteínas do Citoesqueleto , Terapia Genética , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Proteínas com Domínio LIM , Coelhos , Doenças da Coluna Vertebral/fisiopatologia , Doenças da Coluna Vertebral/cirurgia , Fusão Vertebral/tendências , Transfecção
17.
J Bone Joint Surg Am ; 77(9): 1404-17, 1995 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-7673292

RESUMO

The use of a bovine bone-derived osteoinductive protein extract as a bone-graft substitute was evaluated in a rabbit model of intertransverse process arthrodesis of the lumbar spine. Forty-five adult New Zealand White rabbits had arthrodesis between the fifth and sixth lumbar vertebrae with use of one of three graft materials: autogenous iliac-crest bone, osteoinductive protein delivered in an allogeneic demineralized bone matrix/collagen carrier, or demineralized bone matrix/collagen carrier or demineralized bone matrix/collagen carrier without osteoinductive protein. Fusion was assessed by manual palpation, radiography, biomechanical testing, and light microscopy at two and five weeks after the operation. At two weeks, light microscopic analysis of the arthrodesis site in which osteoinductive protein had been used showed that most of the demineralized bone matrix was still present, with small amounts of membranous and endochondral bone formation at the peripheral margins of the implant. Light microscopic analysis of the five-week specimens showed increased new-bone formation and a more homogeneous and mature fusion mass with the osteoinductive bone protein than with the autogenous bone graft. At five weeks, the fusions with the osteoinductive protein extract were characterized by more secondary spongiosa, with formation of bone marrow centrally and a cortical rim peripherally. Of the thirty-five rabbits that were examined at five weeks, all ten in the group that had received osteoinductive bone protein had a solid fusion, but the rate of fusion was significantly less in the other two groups: eight of thirteen rabbits (p = 0.05) in the group that had received autogenous bone graft and two of twelve rabbits (p = 0.0001) in the group that had received demineralized bone matrix/collagen carrier without osteoinductive bone protein. The use of osteoinductive bone protein resulted in stronger (p = 0.02) and stiffer (p = 0.005) fusions compared with those obtained with the use of autogenous iliac-crest graft.


Assuntos
Glicoproteínas/administração & dosagem , Substâncias de Crescimento/administração & dosagem , Vértebras Lombares/cirurgia , Fusão Vertebral , Animais , Matriz Óssea , Transplante Ósseo , Colágeno , Vértebras Lombares/patologia , Osteogênese , Coelhos , Fusão Vertebral/métodos
18.
J Bone Joint Surg Am ; 75(9): 1282-97, 1993 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-8408150

RESUMO

We analyzed the cases of seventy-three patients who were managed over a twenty-year period for rheumatoid involvement of the cervical spine and were followed for a minimum of two years, with an average follow-up of seven years. A neurological deficit did not develop in thirty-one patients (Ranawat et al. Class I) and paralysis developed in the remaining forty-two patients: Class II in eleven and Class III in thirty-one. Of the forty-two patients in whom paralysis developed, thirty-five had operative stabilization. Seven patients were managed with a soft cervical collar because they refused or were medically unable to have the operation; all of the had an increase in the severity of the paralysis. The posterior atlanto-odontoid interval and the diameter of the subaxial sagittal canal measured on the cervical radiographs demonstrated statistically significant correlations with the presence and severity of paralysis. All of the patients who had a Class-III neurological deficit had a posterior atlanto-odontoid interval or diameter of the subaxial canal that was less than fourteen millimeters. In contrast, the anterior atlanto-odontoid interval, which has traditionally been reported, did not correlate with paralysis. The prognosis for neurological recovery following the operation was not affected by the duration of the paralysis but was influenced by the severity of the paralysis at the time of the operation. The most important predictor of the potential for neurological recovery after the operation was the preoperative posterior atlanto-odontoid interval. In patients who had paralysis due to atlanto-axial subluxation, no recovery occurred if the posterior atlanto-odontoid interval was less than ten millimeters, whereas recovery of at least one neurological class always occurred when the posterior atlanto-odontoid interval was at least ten millimeters. If basilar invagination was superimposed, clinically important neurological recovery occurred only when the posterior atlanto-odontoid interval was at least thirteen millimeters. All patients who had paralysis and a posterior atlanto-odontoid interval or diameter of the subaxial canal of fourteen millimeters had complete motor recovery after the operation. In this series, although only patients who had a neurological deficit were operated on, we observed the range of the posterior atlanto-odontoid interval that was associated with poor or no recovery after the operation, and we identified the safe range on the basis of the patients in whom paralysis did not develop.(ABSTRACT TRUNCATED AT 400 WORDS)


Assuntos
Artrite Reumatoide/cirurgia , Vértebras Cervicais , Adulto , Idoso , Idoso de 80 Anos ou mais , Artrite Reumatoide/classificação , Artrite Reumatoide/complicações , Artrite Reumatoide/diagnóstico por imagem , Artrodese , Articulação Atlantoaxial/diagnóstico por imagem , Articulação Atlantoaxial/cirurgia , Vértebras Cervicais/diagnóstico por imagem , Vértebras Cervicais/cirurgia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Paralisia/etiologia , Prognóstico , Radiografia , Estudos Retrospectivos , Doenças da Coluna Vertebral/classificação , Doenças da Coluna Vertebral/diagnóstico por imagem , Doenças da Coluna Vertebral/cirurgia
19.
J Bone Joint Surg Am ; 76(12): 1847-53, 1994 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-7989390

RESUMO

Seventeen fresh segments of cadaveric lumbar spines were tested in flexion, extension, and axial rotation. The resulting angular rotations were measured with the use of a goniometer and a three-dimensional system of video analysis. Measurements of flexibility were made, in order, in the intact spine; after decompression (bilateral total laminectomies, partial medial facetectomies, and foraminotomies); after excision of the capsule and cartilage of the facets; and after cancellous bone had been packed into the facet defects. Decompression resulted in a slight increase in the sagittal and axial ranges of motion. Subsequent excision of the capsule and cartilage of the facets, as in preparation for an arthrodesis of the facets, resulted in a significant increase in both the sagittal (5.7 +/- 2.9 degrees, mean and standard deviation) (p < 0.001) and the axial (1.4 +/- 0.9 degrees) (p < 0.01) ranges of motion compared with the motion in the intact specimen and with the motion in the specimen after only decompression had been done (p < 0.01 and p < 0.05, respectively). Packing of bone in the facets did not significantly reduce motion. It was calculated that the increase in the sagittal range of motion after excision of the capsule and cartilage of the facets would increase the tensile strain in a graft between the transverse processes of the fourth and fifth lumbar vertebrae (18 +/- 1 per cent tensile strain [mean and 95 per cent confidence interval] for the intact vertebrae and 25 +/- 1 per cent for the vertebrae in which the facets had been excised).


Assuntos
Cartilagem Articular/cirurgia , Cápsula Articular/cirurgia , Vértebras Lombares/fisiologia , Fenômenos Biomecânicos , Transplante Ósseo , Humanos , Laminectomia , Vértebras Lombares/cirurgia , Movimento , Amplitude de Movimento Articular , Fusão Vertebral , Resistência à Tração
20.
J Bone Joint Surg Am ; 78(3): 403-11, 1996 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-8613448

RESUMO

The orientation of the lumbar facet joints was studied with magnetic resonance imaging in 140 subjects to determine if there is an association between facet tropism and intervertebral disc disease or between the orientation of the facet joints and degenerative spondylolisthesis. The 140 subjects were divided into four groups: sixty-seven asymptomatic volunteers, forty-six of whom did not have a herniated disc on magnetic resonance scans (Group I) and twenty-one who did (Group II); forty-six symptomatic patients who had a herniated disc confirmed operatively (Group III); and twenty-seven patients who had degenerative spondylolisthesis at the interspace between the fourth and fifth lumbar vertebrae (Group IV). Axial scans were made at each lumbar level and digitized, and the facet joint angle was measured by two independent observers with use of image analysis software in a personal computer. The technique of measurement of the facet angles on magnetic resonance scans was validated with a subset of subjects who also had computed tomography scans made. Similar values were obtained with the two methods (r = 0.92; p = 0.00001). For the forty-six asymptomatic volunteers who did not have a herniated disc on the magnetic resonance scans (Group I), the median facet tropism was 5 to 6 degrees and was more than 10 degrees in 24 per cent (eleven) of the subjects. There was no association between increased facet tropism and disc degeneration. At the level of the fourth and fifth lumbar vertebrae, the median facet tropism was 10.3 degrees in the symptomatic patients who had a herniated disc at the same level and 5.4 degrees in the asymptomatic volunteers (Group I) (p = 0.05). The mean orientation of the lumbar facet angles relative to the coronal plane was more sagittal at all levels in the patients who had degenerative spondylolisthesis. The greatest difference was at the level of the fourth and fifth lumbar vertebrae (p = 0.000001). The mean facet angle was 41 degrees (95 per cent confidence interval, 37.6 to 44.6 degrees) in the asymptomatic volunteers and 60 degrees (95 per cent confidence interval, 52.7 to 67.1 degrees) in the patients who had degenerative spondylolisthesis. Furthermore, both the left and the right facet joints were more sagittally oriented in the patients who had degenerative spondylolisthesis. An individual in who both facet-joint angles at the level of the fourth and fifth lumbar vertebrae were more than 45 degrees relative to the coronal plane was twenty-five times more likely to have degenerative spondylolisthesis (95 per cent confidence interval, seven to ninety-eight times). The increase in facet angles at levels other than that of the spondylolisthesis suggests that increased facet angles represent variations in anatomy rather than a secondary result of spondylolisthesis.


Assuntos
Deslocamento do Disco Intervertebral/patologia , Vértebras Lombares/patologia , Espondilolistese/patologia , Adulto , Humanos , Disco Intervertebral/patologia , Articulações/anatomia & histologia , Articulações/patologia , Vértebras Lombares/anatomia & histologia , Imageamento por Ressonância Magnética , Osteoartrite/patologia , Reprodutibilidade dos Testes
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