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1.
Osteoporos Int ; 27(1): 127-33, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26187124

RESUMO

UNLABELLED: Trabecular bone score (TBS) seems to provide additive value on BMD to identify individuals with prevalent fractures in T1D. TBS did not significantly differ between T1D patients and healthy controls, but TBS and HbA1c were independently associated with prevalent fractures in T1D. A TBS cutoff <1.42 reflected prevalent fractures with 91.7 % sensitivity and 43.2 % specificity. INTRODUCTION: Type 1 diabetes (T1D) increases the risk of osteoporotic fractures. TBS was recently proposed as an indirect measure of bone microarchitecture. This study aimed at investigating the TBS in T1D patients and healthy controls. Associations with prevalent fractures were tested. METHODS: One hundred nineteen T1D patients (59 males, 60 premenopausal females; mean age 43.4 ± 8.9 years) and 68 healthy controls matched for gender, age, and body mass index (BMI) were analyzed. The TBS was calculated in the lumbar region, based on two-dimensional (2D) projections of DXA assessments. RESULTS: TBS was 1.357 ± 0.129 in T1D patients and 1.389 ± 0.085 in controls (p = 0.075). T1D patients with prevalent fractures (n = 24) had a significantly lower TBS than T1D patients without fractures (1.309 ± 0.125 versus 1.370 ± 0.127, p = 0.04). The presence of fractures in T1D was associated with lower TBS (odds ratio = 0.024, 95 % confidence interval (CI) = 0.001-0.875; p = 0.042) but not with age or BMI. TBS and HbA1c were independently associated with fractures. The area-under-the curve (AUC) of TBS was similar to that of total hip BMD in discriminating T1D patients with or without prevalent fractures. In this set-up, a TBS cutoff <1.42 discriminated the presence of fractures with a sensitivity of 91.7 % and a specificity of 43.2 %. CONCLUSIONS: TBS values are lower in T1D patients with prevalent fractures, suggesting an alteration of bone strength in this subgroup of patients. Reliable TBS cutoffs for the prediction of fracture risk in T1D need to be determined in larger prospective studies.


Assuntos
Diabetes Mellitus Tipo 1/complicações , Vértebras Lombares/diagnóstico por imagem , Fraturas por Osteoporose/diagnóstico por imagem , Fraturas por Osteoporose/etiologia , Absorciometria de Fóton/métodos , Adulto , Idoso , Índice de Massa Corporal , Densidade Óssea/fisiologia , Estudos de Casos e Controles , Estudos Transversais , Diabetes Mellitus Tipo 1/fisiopatologia , Feminino , Colo do Fêmur/fisiopatologia , Hemoglobinas Glicadas/metabolismo , Articulação do Quadril/fisiopatologia , Humanos , Vértebras Lombares/fisiopatologia , Masculino , Pessoa de Meia-Idade , Fraturas por Osteoporose/fisiopatologia , Sensibilidade e Especificidade , Adulto Jovem
2.
Osteoporos Int ; 25(5): 1527-33, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24599273

RESUMO

UNLABELLED: Fracture risk in type 1 diabetes (T1D) is supposed to be underestimated by bone mineral density (BMD). Individuals with T1D had more prevalent fractures in a cross-sectional study. Serum levels of pentosidine, an advanced glycation end product, and poor glycaemic control were associated with prevalent fractures independent of BMD. INTRODUCTION: Type 1 diabetes (T1D) is associated with increased fracture risk. Bone mineral density (BMD) underestimates the risk of fractures in some individuals. The accumulation of advanced glycation end products (AGEs) impairs bone matrix and reduces bone strength. METHODS: In a cross-sectional study, 128 men and premenopausal women with T1D were evaluated. We compared traditional risk factors for fractures, BMD, parameters of bone metabolism and AGEs in individuals with and without prevalent fractures. An independent association of serum AGE levels with prevalent fractures was investigated. RESULTS: Individuals with prevalent fractures exhibited a longer duration of T1D, higher HbA1c and more diabetic-related complications. BMD at the femoral neck (z-score -0.76 ± 0.94 vs. -0.23 ± 1.02; p = 0.031) and total hip (z-score -0.54 ± 0.93 vs. 0.11 ± 1.11; p = 0.017) was lower in those with prevalent fractures. Individuals with fractures had higher pentosidine levels (164.1 ± 53.6 vs. 133.2 ± 40.4; p = 0.002). The levels of N-ε-(carboxymethyl)-lysine (CML) and endogenous secretory receptor for AGEs (esRAGE) did not significantly differ. Multivariate logistic regression analysis adjusted for age, BMI, family history of fractures, smoking, vitamin D deficiency, BMD at lumbar spine, femoral neck and total hip identified pentosidine levels and HbA1c as independent factors associated with prevalent fractures (odds ratio 1.02, 95% CI 1.00-1.03/pmol/ml increase of pentosidine; p = 0.008 and odds ratio 1.93, 95% CI 1.16-3.20 per percentage increase of HbA1c; p = 0.011). CONCLUSIONS: The pentosidine levels but not BMD are independently associated with prevalent fractures. Impaired bone quality in T1D may result from increased AGE formation.


Assuntos
Arginina/análogos & derivados , Densidade Óssea/fisiologia , Diabetes Mellitus Tipo 1/complicações , Lisina/análogos & derivados , Fraturas por Osteoporose/etiologia , Receptores Imunológicos/sangue , Adulto , Arginina/sangue , Biomarcadores/sangue , Glicemia/metabolismo , Estudos Transversais , Diabetes Mellitus Tipo 1/sangue , Diabetes Mellitus Tipo 1/fisiopatologia , Feminino , Hemoglobinas Glicadas/metabolismo , Humanos , Lisina/sangue , Masculino , Pessoa de Meia-Idade , Fraturas por Osteoporose/sangue , Fraturas por Osteoporose/fisiopatologia , Receptor para Produtos Finais de Glicação Avançada , Medição de Risco/métodos
3.
Diabet Med ; 28(7): 872-5, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21395677

RESUMO

AIM: There are conflicting data regarding the risk of osteoporosis in patients with Type 1 diabetes. We investigated an association between diabetes, bone mineral density and prevalent fractures. METHODS: A single-centre, cross-sectional study of men and pre-menopausal women with Type 1 diabetes (n = 128) and a matched control group (n = 77) was conducted. The primary outcome measure was bone mineral density and secondary measures were markers of bone metabolism and prevalent fractures. RESULTS: Hip and total body bone mineral densities were significantly lower in women with diabetes compared with control subjects. In men, no difference in bone mineral density was found. A multivariate regression analysis in women with diabetes revealed higher BMI as the strongest predictor of higher total hip, femoral neck and total body bone mineral density, whereas previous fractures were inversely associated with total hip bone mineral density and C-terminal telopeptide of type I collagen with total body bone mineral density. Poor long-term glycaemic control was not associated with low bone mineral density. Fracture frequency was higher in patients with diabetes compared with control subjects (1.64 vs. 0.62 per 100 patient-years; P < 0.05). In a multivariable model, long-term HbA(1c) control was associated with increased clinical fracture prevalence (OR 1.92; 95% CI 1.09-2.75) in those with diabetes. CONCLUSIONS: Type 1 diabetes contributes to low bone mineral density in women. Previous fractures and low BMI were strong predictors of impaired bone mineral density and should therefore be considered in risk estimation. Fractures are more frequent in Type 1 diabetes. Long-term hyperglycaemia may account for impaired bone strength, independently from bone mineral density.


Assuntos
Densidade Óssea/fisiologia , Diabetes Mellitus Tipo 1/fisiopatologia , Fraturas Ósseas/fisiopatologia , Vértebras Lombares/fisiopatologia , Osteoporose/fisiopatologia , Biomarcadores/metabolismo , Densidade Óssea/efeitos dos fármacos , Estudos de Casos e Controles , Estudos Transversais , Diabetes Mellitus Tipo 1/complicações , Diabetes Mellitus Tipo 1/tratamento farmacológico , Feminino , Fraturas Ósseas/diagnóstico por imagem , Fraturas Ósseas/metabolismo , Hemoglobinas Glicadas/metabolismo , Humanos , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/metabolismo , Masculino , Pessoa de Meia-Idade , Osteoporose/induzido quimicamente , Osteoporose/metabolismo , Prevalência , Radiografia , Fatores de Risco
4.
J Cell Biol ; 116(4): 839-49, 1992 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-1531145

RESUMO

We have investigated the structure of the small nuclear RNP (snRNP) U1 by combining EM of complete and partially protein-deficient particles with immunoelectron microscopy employing mAbs against known components of the U1 snRNP. It was found that the two main protuberances of this particle can be identified with the U1-specific proteins A and 70K. The 70K protuberance is the one lying closer to the 5' terminus of the snRNA, as identified by its 5'-terminal m3G cap. The round-shaped main body of U1 snRNP represents its core RNP domain containing the common snRNP proteins. Functional implications of these results are discussed. Our results may also point to the physical basis for the production of autoantibodies directed against specific groups of snRNP proteins. The physical grouping of the common proteins (Sm epitopes) and the specific proteins (RNP epitopes) could result in one or the other being presented to the immune system as is the case in patients suffering from SLE or MCTD, respectively.


Assuntos
Ribonucleoproteínas/química , Especificidade de Anticorpos , Complexo Antígeno-Anticorpo , Reagentes de Ligações Cruzadas , Células HeLa , Humanos , Microscopia Imunoeletrônica , Ribonucleoproteínas/imunologia , Ribonucleoproteínas/ultraestrutura , Ribonucleoproteínas Nucleares Pequenas , Succinimidas
5.
J Cell Biol ; 112(6): 1065-72, 1991 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-1825657

RESUMO

We describe the electron microscopic investigation of purified U4/U6 snRNPs from human and murine cells. The U4/U6 snRNP exhibits two morphological features, a main body approximately 8 nm in diameter and a peripheral filamentous domain, 7-10 nm long. Two lines of evidence suggest that the peripheral domain may consist of RNA and to contain U6 RNA as well as the 5' portion of U4 RNA. (a) Separation of the U4/U6 snRNA interaction regions from the core domains by site-directed cleavage of the U4 snRNA with RNase H gave filament-free, globular core snRNP structures. (b) By immuno and DNA-hybridization EM, both the 5' end of U4 and the 3' end of U6 snRNA were located at the distal region of the filamentous domain, furthest from the core. These results, together with our observation that the filamentous U4/U6 domain is often Y shaped, correlate strikingly with the consensus secondary structure proposed by Brow and Guthrie (1988. Nature (Lond.), 334:213-218), where U4 and U6 snRNA are base paired in such a way that two U4/U6 helices together with a stem/loop of U4 snRNA make up a Y-shaped U4/U6 interaction domain.


Assuntos
RNA Nuclear Pequeno/ultraestrutura , Ribonucleoproteínas/ultraestrutura , Animais , Anticorpos Monoclonais , Complexo Antígeno-Anticorpo , Sequência de Bases , Carcinoma de Ehrlich/metabolismo , Células HeLa/metabolismo , Humanos , Camundongos , Microscopia Eletrônica , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Sondas de Oligonucleotídeos , Ribonucleoproteínas/imunologia , Ribonucleoproteínas/isolamento & purificação , Ribonucleoproteínas Nucleares Pequenas
6.
J Cell Biol ; 145(7): 1355-68, 1999 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-10385517

RESUMO

The 17S U2 small nuclear ribonucleoprotein particle (snRNP) represents the active form of U2 snRNP that binds to the pre-mRNA during spliceosome assembly. This particle forms by sequential interactions of splicing factors SF3b and SF3a with the 12S U2 snRNP. We have purified SF3b and the 15S U2 snRNP, an intermediate in the assembly pathway, from HeLa cell nuclear extracts and show that SF3b consists of four subunits of 49, 130, 145, and 155 kD. Biochemical analysis indicates that both SF3b and the 12S U2 snRNP are required for the incorporation of SF3a into the 17S U2 snRNP. Nuclease protection studies demonstrate interactions of SF3b with the 5' half of U2 small nuclear RNA, whereas SF3a associates with the 3' portion of the U2 snRNP and possibly also interacts with SF3b. Electron microscopy of the 15S U2 snRNP shows that it consists of two domains in which the characteristic features of isolated SF3b and the 12S U2 snRNP are conserved. Comparison to the two-domain structure of the 17S U2 snRNP corroborates the biochemical results in that binding of SF3a contributes to an increase in size of the 12S U2 domain and possibly induces a structural change in the SF3b domain.


Assuntos
Ribonucleoproteína Nuclear Pequena U2/ultraestrutura , Spliceossomos/química , Resinas de Troca Aniônica , Sequência de Bases , Cromatografia Líquida , Células HeLa , Humanos , Nuclease do Micrococo , Microscopia Eletrônica , Modelos Moleculares , Dados de Sequência Molecular , Proteínas Nucleares/análise , Proteínas Nucleares/isolamento & purificação , Proteínas Nucleares/metabolismo , Conformação de Ácido Nucleico , Fosfoproteínas/química , Fosfoproteínas/isolamento & purificação , Fosfoproteínas/metabolismo , Fosfoproteínas/ultraestrutura , Testes de Precipitina , Ligação Proteica , Conformação Proteica , Fatores de Processamento de RNA , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/isolamento & purificação , Proteínas de Ligação a RNA/metabolismo , Resinas Sintéticas , Ribonucleoproteína Nuclear Pequena U2/química , Ribonucleoproteína Nuclear Pequena U2/isolamento & purificação , Ribonucleoproteína Nuclear Pequena U2/metabolismo , Spliceossomos/metabolismo , Spliceossomos/ultraestrutura
7.
Science ; 264(5156): 261-5, 1994 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-8146658

RESUMO

Small nuclear ribonucleoprotein (snRNP) particles are essential for pre-messenger RNA splicing. In human HeLa cells, 40 proteins associated with snRNPs have been identified. Yet, the function of many of these proteins remains unknown. Here, the immunoaffinity purification of the spliceosomal snRNPs U1, U2, U4/U6.U5, and several nucleolar snRNP species from the yeast Saccharomyces cerevisiae is presented. The U1 and U4/U6.U5 snRNPs were purified extensively and their protein composition and ultrastructure analyzed. The yeast U1 snRNP is larger and contains three times more specific proteins than its human counterpart. In contrast, the size, protein composition, and morphology of the yeast and the human U4/U6.U5 snRNPs are significantly similar. The preparative isolation of yeast snRNPs will allow the cloning as well as genetic and phylogenetic analysis of snRNP proteins which will accelerate our understanding of their function.


Assuntos
Ribonucleoproteínas Nucleares Pequenas/isolamento & purificação , Saccharomyces cerevisiae/química , Spliceossomos/química , Western Blotting , Centrifugação com Gradiente de Concentração , Cromatografia de Afinidade , Células HeLa , Humanos , Microscopia Eletrônica , Peso Molecular , RNA Fúngico/análise , Ribonucleoproteína Nuclear Pequena U1/química , Ribonucleoproteína Nuclear Pequena U1/isolamento & purificação , Ribonucleoproteína Nuclear Pequena U4-U6/química , Ribonucleoproteína Nuclear Pequena U4-U6/isolamento & purificação , Ribonucleoproteína Nuclear Pequena U5/química , Ribonucleoproteína Nuclear Pequena U5/isolamento & purificação , Ribonucleoproteínas Nucleares Pequenas/química
8.
J Phys Condens Matter ; 20(37): 374108, 2008 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-21694415

RESUMO

Two strongly coupled quantum dots are theoretically and experimentally investigated. In conductance measurements on a GaAs based low-dimensional system additional features to the Coulomb blockade have been detected at low temperatures. These regions of finite conductivity are compared with theoretical investigations of a strongly coupled quantum dot system and good agreement between the theoretical and the experimental results has been found.

9.
Mol Cell Biol ; 19(10): 6554-65, 1999 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-10490595

RESUMO

The association of Sm proteins with U small nuclear RNA (snRNA) requires the single-stranded Sm site (PuAU(4-6)GPu) but also is influenced by nonconserved flanking RNA structural elements. Here we demonstrate that a nonameric Sm site RNA oligonucleotide sufficed for sequence-specific assembly of a minimal core ribonucleoprotein (RNP), which contained all seven Sm proteins. The minimal core RNP displayed several conserved biochemical features of native U snRNP core particles, including a similar morphology in electron micrographs. This minimal system allowed us to study in detail the RNA requirements for Sm protein-Sm site interactions as well as the kinetics of core RNP assembly. In addition to the uridine bases, the 2' hydroxyl moieties were important for stable RNP formation, indicating that both the sugar backbone and the bases are intimately involved in RNA-protein interactions. Moreover, our data imply that an initial phase of core RNP assembly is mediated by a high affinity of the Sm proteins for the single-stranded uridine tract but that the presence of the conserved adenosine (PuAU.) is essential to commit the RNP particle to thermodynamic stability. Comparison of intact U4 and U5 snRNAs with the Sm site oligonucleotide in core RNP assembly revealed that the regions flanking the Sm site within the U snRNAs facilitate the kinetics of core RNP assembly by increasing the rate of Sm protein association and by decreasing the activation energy.


Assuntos
Oligorribonucleotídeos/metabolismo , RNA Nuclear Pequeno/metabolismo , Proteínas de Ligação a RNA/metabolismo , Ribonucleoproteínas Nucleares Pequenas/metabolismo , Spliceossomos/metabolismo , Sítios de Ligação , Centrifugação com Gradiente de Concentração , Células HeLa , Humanos , Cinética , Ligação Proteica , Ribonucleoproteínas Nucleares Pequenas/ultraestrutura
10.
Mol Cell Biol ; 13(1): 307-19, 1993 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-8380223

RESUMO

Small nuclear (sn) ribonucleoprotein (RNP) U2 functions in the splicing of mRNA by recognizing the branch site of the unspliced pre-mRNA. When HeLa nuclear splicing extracts are centrifuged on glycerol gradients, U2 snRNPs sediment at either 12S (under high salt concentration conditions) or 17S (under low salt concentration conditions). We isolated the 17S U2 snRNPs from splicing extracts under nondenaturing conditions by using centrifugation and immunoaffinity chromatography and examined their structure by electron microscope. In addition to common proteins B', B, D1, D2, D3, E, F, and G and U2-specific proteins A' and B", which are present in the 12S U2 snRNP, at least nine previously unidentified proteins with apparent molecular masses of 35, 53, 60, 66, 92, 110, 120, 150, and 160 kDa bound to the 17S U2 snRNP. The latter proteins dissociate from the U2 snRNP at salt concentrations above 200 mM, yielding the 12S U2 snRNP particle. Under the electron microscope, the 17S U2 snRNPs exhibited a bipartite appearance, with two main globular domains connected by a short filamentous structure that is sensitive to RNase. These findings suggest that the additional globular domain, which is absent from 12S U2 snRNPs, contains some of the 17S U2-specific proteins. The 5' end of the RNA in the U2 snRNP is more exposed for reaction with RNase H and with chemical probes when the U2 snRNP is in the 17S form than when it is in the 12S form. Removal of the 5' end of this RNA reduces the snRNP's Svedberg value from 17S to 12S. Along with the peculiar morphology of the 17S snRNP, these data indicate that most of the 17S U2-specific proteins are bound to the 5' half of the U2 snRNA.


Assuntos
Splicing de RNA , Ribonucleoproteína Nuclear Pequena U2/química , Sequência de Bases , Fracionamento Celular/métodos , Núcleo Celular/química , Células HeLa , Humanos , Ligação de Hidrogênio , Técnicas In Vitro , Microscopia Eletrônica , Dados de Sequência Molecular , Peso Molecular , Conformação de Ácido Nucleico , Proteínas de Ligação a RNA/química , Ribonuclease H/farmacologia
11.
Bone ; 82: 50-5, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25888930

RESUMO

To determine osteocalcin (OC) and adipokines in type 1 diabetes (T1D) and healthy controls, and to explore possible associations between glucose and bone metabolism, body composition and adipokines. Serum levels of total OC, undercarboxylated (UC-OC), leptin, adiponectin, and other parameters of glucose and bone metabolism were measured in 128 patients with T1D (mean duration 21.2years) and in 77 healthy controls, matched for gender, age, and body mass index (BMI). Partial correlations (adjusted for age and gender) with parameters of body composition (BMI, fat body mass [derived from bone mineral density scans]), glycaemic control (hemoglobin A1c (HbA1c), daily insulin dose in T1D), skeletal homeostasis (osteoprotegerin (OPG), receptor activator of NF-κB ligand (RANKL), all measured in serum), and serum insulin-like growth factor 1 (IGF-1) were also examined. Independent predictors of total and UC-OC were then explored. Total OC was lower in males with T1D (16.3±6.4 vs. 22.2±9.9ng/ml; p=0.001), whereas UC-OC did not show group differences. Adiponectin was higher in T1D patients, both for males and females (8.9±6.6 vs. 5.7±2.5µg/ml; p=0.004 and 13.8±6.4 vs. 8.8±4.0µg/ml; p<0.001). IGF-1 was lower only in females with T1D (146.6±68.8 vs. 203.0±74.4ng/ml; p<0.001). BMI and fat body mass were similar in T1D and controls. In T1D patients, total OC was inversely correlated with BMI and HbA1c, and UC-OC inversely correlated with HbA1c. In T1D patients, leptin positively correlated with BMI, fat body mass and daily insulin dose, while adiponectin inversely correlated with BMI and daily insulin dose. Multivariate regression modelling showed that determinants of higher total OC levels were male gender (p=0.04, ß-coefficient=2.865) and lower HbA1c (p=0.04, ß-coefficient=-0.117), whereas determinants of UC-OC levels were T1D (p=0.016, ß-coefficient=2.015), higher IGF-1 (p=0.004, ß-coefficient=0.011) and lower HbA1c (p=0.011, ß-coefficient=- 0.061). Total OC and UC-OC are associated with good glycaemic control in T1D, with gender-specific differences for total-OC. The association of leptin and adiponectin with glycaemic control, as observed in controls, does not seem to be a feature in T1D, although both adipokines appear to be related to the insulin demand. This article is part of a Special Issue entitled "Bone and diabetes".


Assuntos
Adipocinas/sangue , Glicemia/metabolismo , Diabetes Mellitus Tipo 1/sangue , Diabetes Mellitus Tipo 1/diagnóstico , Osteocalcina/sangue , Adulto , Idoso , Biomarcadores/sangue , Biomarcadores/metabolismo , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores Sexuais , Adulto Jovem
12.
J Mol Biol ; 265(2): 87-94, 1997 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-9020971

RESUMO

All four spliceosomal small nuclear ribonucleoproteins (snRNPs) U1, U2, U4/U6 and U5 contain a common structural element called the snRNP core. This core is assembled from the common snRNP proteins and the small nuclear RNA (snRNA). We have used electron microscopy to study the structure of two intermediates of the snRNP core assembly pathway: (1) the (E.F.G) protein complex, which contains only the smallest common proteins E, F and G; and (2) the subscore of U5 snRNP, in which the U5 RNA and the common proteins D1 and D2 are bound to the (E.F.G) protein complex. The general structure of the subscore was found to resemble that of the complete snRNP core, which contains the components of the subscore plus the common proteins B/B' and D3. Both the complete snRNP core and subscore particles are globular, with diameters of 7 to 8 nm. They show a characteristic accumulation of stain at the centre. However, some subscore images showed nicked outlines not seen with the complete snRNP cores. The (E.F.G) protein complex appeared as a ring, with an outer diameter of about 7 nm and a central hole 2 nm across. The molecular dimensions of the E, F and G proteins imply that the thickness of the (E.F.G) ring structure is only about 2 nm. Comparison of the (E.F.G) structure complex with the snRNP core and subcore structures implicates that a flat side of the ring-shaped (E.F.G) complex provides the assembly site(s) for the other components of the snRNP during core assembly: first for the D1 and D2 proteins (and probably the snRNA) during subscore formation, and then for the B/B' and D3 proteins in the completion of the snRNP core particle.


Assuntos
RNA Nuclear Pequeno/ultraestrutura , Ribonucleoproteínas Nucleares Pequenas/ultraestrutura , Centrifugação com Gradiente de Concentração , Células HeLa , Humanos , Microscopia Eletrônica , Conformação Proteica , RNA Nuclear Pequeno/metabolismo , Ribonucleoproteína Nuclear Pequena U1/metabolismo , Ribonucleoproteína Nuclear Pequena U1/ultraestrutura , Ribonucleoproteína Nuclear Pequena U2/genética , Ribonucleoproteína Nuclear Pequena U2/metabolismo , Ribonucleoproteína Nuclear Pequena U5/metabolismo , Ribonucleoproteína Nuclear Pequena U5/ultraestrutura , Ribonucleoproteínas Nucleares Pequenas/metabolismo , Spliceossomos
13.
J Mol Biol ; 212(2): 241-5, 1990 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-2181146

RESUMO

In protein synthesis Escherichia coli release factor-2 binds to 70 S ribosomes when the termination codon UAA or UGA appears at the decoding site. The weak interaction between factor and ribosome has been stabilized in vitro by chemical cross-linking. Factor so bound can still be recognized by a specific antibody to release factor-2. Examination of the resulting immuno-complexes by electron microscopy revealed 70 S ribosomes in different projection forms, and the occasional dissociated subunit labelled with antibody. The antibody-binding site was localized on previously characterized 70 S projection forms, and its three-dimensional localization on the 70 S model established. The release factor-2-binding site was found to be positioned at the ribosomal subunit interface, comprising the stalk-protuberance region of the large subunit and the head-neck region of the concave side of the small subunit.


Assuntos
Escherichia coli/metabolismo , Fatores de Terminação de Peptídeos/metabolismo , Ribossomos/metabolismo , Sítios de Ligação , Imuno-Histoquímica , Microscopia Eletrônica , Ribossomos/ultraestrutura
14.
Pediatrics ; 105(3 Pt 1): 496-501, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10699099

RESUMO

BACKGROUND: Clinical practice guidelines are increasingly being used for a wide variety of medical conditions, but not enough is known about physicians' attitudes and beliefs about guidelines, how often and under what circumstances they are used, and factors associated with their acceptance. OBJECTIVE: To determine practice guideline attitudes, beliefs, practices, and factors associated with use among a representative national sample of general pediatricians. STUDY DESIGN: Cross-sectional mail survey. SUBJECTS: Random sample of general pediatrician members of the American Academy of Pediatrics residing in all 50 states and Puerto Rico. SURVEY INSTRUMENT: Twenty-four multiple-choice, Likert scale, yes-no, and open-ended questions about pediatric clinical practice guidelines. RESULTS: From 1088 respondents, 461 specialists were excluded; the remaining 627 general pediatricians were mostly male (61%), white (81%), and in group practice (62%) in a suburban location (48%). Practice guidelines are used by 35% of pediatricians, in part by 44%, and not at all by 21%. Over 100 different practice guidelines are used, most commonly for asthma (77%), hyperbilirubinemia (27%), and otitis media (19%). Common reasons for use of practice guidelines include standardization of care (17%) and helpfulness (10%). Commonly cited problems with practice guidelines include failure to allow for clinical judgment (54%), use in litigation (16%), and limitation of autonomy (5%). In multivariate analysis, the odds of practice guideline use were greater among pediatricians in health maintenance organization practices (odds ratio [OR]: 9.1; 95% confidence interval [CI]: 1.2-68.0) and those who were nonwhite (OR: 2.3; 95% CI: 1.1-4.8), but lower in those with more weekly patient visits (OR:.7; 95% CI:.5-.9). Features most likely to lead to practice guideline use include simplicity (16%), feasibility (12%), and evidence of improved outcomes (10%). Most pediatricians agree that practice guidelines improve outcomes (89%), are motivated by a desire to improve quality (94%), and should not be used in litigation (82%) or disciplinary actions (77%), nor be motivated by a desire to reduce costs (73%). CONCLUSIONS: Most general pediatricians use practice guidelines, but no specific guidelines, except those for asthma, are used by >27% of pediatricians. The results of this study suggest that practice guidelines are most likely to be followed if they are simple, flexible, rigorously tested, not used punitively, and are motivated by desires to improve quality, not reduce costs.


Assuntos
Atitude do Pessoal de Saúde , Pediatria/estatística & dados numéricos , Guias de Prática Clínica como Assunto , Padrões de Prática Médica/estatística & dados numéricos , Criança , Estudos Transversais , Coleta de Dados , Feminino , Sistemas Pré-Pagos de Saúde/estatística & dados numéricos , Pesquisa sobre Serviços de Saúde/estatística & dados numéricos , Humanos , Masculino , Garantia da Qualidade dos Cuidados de Saúde/estatística & dados numéricos , Estados Unidos
15.
Pediatrics ; 106(1 Pt 2): 231-3, 2000 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10888698

RESUMO

OBJECTIVE: To compare daytime nurse telephone triage calls received by a suburban practice with those received by a hospital-based, inner-city pediatric practice. METHODS: A research assistant, listening simultaneously with triage personnel, prospectively coded all calls received by the nurse triage telephone offices in 2 pediatric practices. Calls were coded the first full, nonholiday week of 3 consecutive summer and 3 consecutive winter months, alternating morning and afternoon sessions. One practice was suburban and had almost all commercially insured patients; the other was hospital-based, in an inner-city, and consisted mostly of patients with Medicaid coverage or no health insurance. RESULTS: A total of 901 calls were triaged in the suburban practice (SP) and 768 in the urban practice (UP). The chief complaints of calls regarding medical problems were similar at both sites. Difficulties with language were noted less often in SP compared with UP (1% vs 17%). The reason for the calls differed by site: medical problem relating to illness or injury (SP 55% vs UP 40%); social issue (SP 1% vs UP 9%); documentation request (SP 2% vs UP 7%); request for laboratory work (SP.3% vs UP 4%); and well child advice (SP 9% vs UP 5%). The disposition of calls also differed by site: telephone advice was offered significantly more in SP than in UP (32% vs 20%); fewer calls required the need for the medical record in SP than in UP (2% vs 12%). CONCLUSIONS: Calls received by a daytime nurse telephone triage office in an affluent SP and a UP are similar in regard to medical problems. Training programs can feasibly prepare physicians and nurses for both kinds of practice settings. The urban site received more calls affected by language and social issues. This could have administrative implications for staffing ratios, language skills of staff and knowledge of available support services.


Assuntos
Pediatria , Saúde Suburbana , Telefone , Triagem/estatística & dados numéricos , Saúde da População Urbana , Criança , Barreiras de Comunicação , Humanos , Enfermeiras e Enfermeiros , Fatores de Tempo
16.
Arch Pediatr Adolesc Med ; 155(6): 699-703, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11386961

RESUMO

OBJECTIVES: To describe patients with nontraumatic dental problems treated in our pediatric emergency department (PED) and to determine if barriers to access prompted seeking care in the PED rather than from a dentist or dental clinic. DESIGN: Questionnaire administered to a convenience sample of patients with nontraumatic dental complaints. SETTING: An urban PED. MAIN OUTCOME MEASURES: Insurance status, primary medical and dental care, duration of symptoms, diagnosis, and reason for seeking care in the PED. RESULTS: Two hundred patients were enrolled. Median age was 17 years (range, 1-22 years). Forty-five percent were African American. Forty-nine percent had Medicaid. Fifty percent identified a regular dentist, whereas 71% had a primary care physician. Thirty-four percent of patients 4 years and older had not seen a dentist in more than a year. Children younger than 13 years were more likely than teenagers to identify a regular dentist (odds ratio [OR] = 2.8; 95% confidence interval [CI], 1.3-6.1). Those with a regular medical provider were more likely to have a regular dentist (OR = 7.7; 95% CI, 3.4-18). The most common reasons for not going to a dentist were as follows: dentist closed, 34%; lack of dental insurance or money, 17%; and lack of a dentist, 16%. Patients with symptoms for more than 72 hours were more likely to cite lack of a dentist as their reason for coming to the PED (OR = 7.4; 95% CI, 1.9-33). CONCLUSIONS: Many pediatric patients do not have regular dental care, and this is associated with a lack of primary medical care. Access barriers to acute dental care include lack of insurance or funds, lack of a dentist, and limited hours of dental care sites. Improved insurance reimbursement, active enrollment of adolescents into preventive dental care, and expansion of provider hours may limit PED dental visits and improve the health of patients.


Assuntos
Serviço Hospitalar de Emergência , Doenças Dentárias/terapia , Adolescente , Adulto , Distribuição de Qui-Quadrado , Criança , Pré-Escolar , Intervalos de Confiança , Feminino , Acessibilidade aos Serviços de Saúde , Humanos , Lactente , Masculino , Razão de Chances , Inquéritos e Questionários
17.
Arch Pediatr Adolesc Med ; 155(12): 1335-9, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11732952

RESUMO

OBJECTIVE: Physicians providing emergency department care to children primarily use nebulizers for the delivery of bronchodilators and these physicians have misconceptions regarding the advantages and disadvantages of using metered-dose inhalers (MDIs) with a spacer (MDI + S) for acute asthma exacerbations. DESIGN: Self-administered mail survey. SETTING: Emergency department. PARTICIPANTS: Emergency medicine section members of the American Academy of Pediatrics and Canadian Pediatric Society. INTERVENTIONS: Bronchodilator delivery methods in acute pediatric asthma. MAIN OUTCOME MEASURES: The 2 principal outcomes for bivariate analysis were self-reported nebulizer use in all patients and MDI + S use in patients with mild acute asthma. RESULTS: Of eligible physicians, 333 (51%) of 567 responded. The majority were dual trained in pediatrics and pediatric emergency medicine (72%) and practiced full time (83%) in an urban (83%) pediatric emergency department (80%). The most commonly cited advantages of MDIs were their cost (33%) and speed of use (28%). The most commonly cited disadvantages were patient or parent dissatisfaction (24%) and relative ease of nebulizer use (23%). Only 10% to 21% of participants used MDIs in the emergency department and reserved this delivery method for children with mild asthma exacerbations. There were no significant associations between selected respondent demographic variables and the use of MDIs. CONCLUSIONS: Misconceptions regarding the efficacy and safety of MDI + S for the treatment of acute asthma exacerbations exist but are limited to a minority of surveyed emergency medicine physicians caring for children. Nebulizers remain the preferred method of routine bronchodilator delivery by physicians providing care to pediatric asthmatics in the emergency department.


Assuntos
Asma/tratamento farmacológico , Broncodilatadores/administração & dosagem , Serviço Hospitalar de Emergência , Nebulizadores e Vaporizadores , Pediatria , Doença Aguda , Atitude do Pessoal de Saúde , Broncodilatadores/uso terapêutico , Competência Clínica , Pesquisas sobre Atenção à Saúde , Humanos , Padrões de Prática Médica
18.
Arch Pediatr Adolesc Med ; 155(6): 680-2, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11386957

RESUMO

BACKGROUND: Nonoccupational human immunodeficiency virus (HIV) postexposure prophylaxis (PEP) for adults has been described, although the Centers for Disease Control and Prevention, Atlanta, Ga, offer no specific recommendations. There is limited information about its use in children and adolescents. OBJECTIVE: To describe the current practices of physicians in pediatric infectious disease (PID) and pediatric emergency medicine (PEM) departments regarding nonoccupational HIV PEP for children and adolescents. DESIGN: Survey. PARTICIPANTS: Directors of all PID and PEM departments with fellowship programs in the United States and Canada between July and November 1998. MAIN OUTCOME MEASURES: General questions regarding HIV PEP and questions concerning 2 scenarios (5-year-old with a needlestick injury and a 15-year-old after sexual assault). RESULTS: The return rate was 67 (78%) of 86 for PID and 36 (75%) of 48 for PEM physicians. Fewer than 20% of physicians reported institutional policies for nonoccupational HIV PEP; 33% had ever initiated nonoccupational HIV PEP. In both scenarios, PID physicians were more likely than PEM physicians to recommend or offer HIV PEP in the first 24 hours after the incident (55 [83%] of 66 vs 20 [56%] of 36 for needlestick injuries [odds ratio, 4.0; 95% confidence interval, 1.6-10.1] and 47 [72%] of 65 vs 16 [50%] of 32 for sexual assault [odds ratio, 2.6; 95% confidence interval, 1.1-6.3]). Seven different antiretroviral agents in single, dual, or triple drug regimens administered for 2 to 12 weeks were suggested. CONCLUSIONS: Although few physicians reported institutional policies, and only one third had ever initiated HIV PEP, many would offer or recommend HIV PEP for children and adolescents within 24 hours after possible HIV exposure. A wide variation of regimens have been suggested. There is a need for a national consensus for nonoccupational HIV PEP.


Assuntos
Fármacos Anti-HIV/uso terapêutico , Abuso Sexual na Infância , Infecções por HIV/prevenção & controle , HIV-1 , Ferimentos Penetrantes Produzidos por Agulha , Padrões de Prática Médica , Adolescente , Distribuição de Qui-Quadrado , Criança , Pré-Escolar , Feminino , Infecções por HIV/transmissão , Humanos , Masculino , Inquéritos e Questionários
19.
Arch Pediatr Adolesc Med ; 152(11): 1119-25, 1998 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-9811291

RESUMO

BACKGROUND: Latinos will soon be the largest minority group in the United States, but too little is known about major access barriers to health care for this group and whether these barriers result in adverse consequences. OBJECTIVE: To identify important access barriers to health care for Latino children, as cited by parents. DESIGN: Cross-sectional survey of parents of all 203 children coming to the pediatric Latino clinic at an innercity hospital. Questions focused on barriers to health care experienced prior to receiving care at the Latino clinic. RESULTS: Parental ethnicity included Dominican (36%), Puerto Rican (34%), Central American (13%), and South American (11%). Only 42% of parents were American citizens, whereas 36% had green cards, and 13% had no documentation. Eight percent of parents and 65% of the children were born in the United States. Parents rated their ability to speak English as follows: very well/well, 27%; not very well, 46%; and not at all, 26%. The median annual household income was $11,000; 40% of parents never graduated from high school, and 49% headed single-parent households. Forty-three percent of the children were uninsured. A sick child was routinely brought to hospital clinics by 56% of parents, to the emergency department by 21%, and to neighborhood health centers by 21%. When asked to name the single greatest barrier to health care for their children, parents cited language problems (26%), long waiting time at the physician's office (15%), no medical insurance (13%), and difficulty paying medical bills (7%). When parents were asked if a particular barrier had ever caused them not to bring their children in, transportation was cited by 21%; not being able to afford health care, 18%; excessive waiting time in the clinic, 17%; no health insurance, 16%; and lack of cultural understanding by staff, 11%. Some parents who spoke little or no English reported that medical staff not speaking Spanish had led to adverse health consequences for their children, including poor medical care (8%), misdiagnosis (6%), and prescription of inappropriate medications (5%). Multivariate analyses of selected health outcomes using 7 independent variables showed that low family income was significantly associated with greater odds of a child's having suboptimal health status (odds ratio, 1.5; 95% confidence interval, 1.04-2.2) and an increased number of physician visits in the past year (P<.04), but reduced odds (odds ratio, 0.6; 95% confidence interval, 0.4-0.9) of the child's being brought to the emergency department for a routine sick visit. Children whose parents had resided in the United States for fewer than 8 years were at reduced odds (odds ratio, 0.5; 95% confidence interval, 0.2-0.9) for having spent a day or more in bed for illness in the past year. CONCLUSIONS: Parents identified language problems, cultural differences, poverty, lack of health insurance, transportation difficulties, and long waiting times as the major access barriers to health care for Latino children. Language problems can result in adverse health consequences for some children, including poor medical care, misdiagnosis, and inappropriate medication and hospitalization. Low family income is an important independent risk factor among Latino children for suboptimal health and high utilization of health services.


Assuntos
Serviços de Saúde da Criança/estatística & dados numéricos , Acessibilidade aos Serviços de Saúde/estatística & dados numéricos , Hispânico ou Latino/estatística & dados numéricos , Adulto , Boston/epidemiologia , Criança , Barreiras de Comunicação , Estudos Transversais , Características Culturais , Feminino , Pesquisas sobre Atenção à Saúde , Humanos , Modelos Logísticos , Masculino , Pobreza , Fatores Socioeconômicos
20.
Acad Med ; 75(5): 451-5, 2000 May.
Artigo em Inglês | MEDLINE | ID: mdl-10824769

RESUMO

PURPOSE: Despite the importance of culture in health care and the rapid growth of ethnic diversity in the United States and Canada, little is known about the teaching of cultural issues in medical schools. The study goals, therefore, were to determine the number of U.S. and Canadian medical schools that have courses on cultural issues, and to examine the format, content, and timing of those courses. METHOD: The authors contacted the deans of students and/ or directors of courses on cultural issues at all 126 U.S. and all 16 Canadian medical schools. Using a cross-sectional telephone survey, they asked whether each school had a course on cultural sensitivity or multicultural issues and, if so, whether it was separate or contained within a larger course, when in the curriculum the course was taught, and which ethnic groups the course addressed. RESULTS: The response rates were 94% for both U.S. (118) and Canadian (15) schools. Very few schools (U.S. = 8%; and Canada = 0%) had separate courses specifically addressing cultural issues. Schools in both countries usually addressed cultural issues in one to three lectures as part of larger, mostly preclinical courses. Significantly more Canadian than U.S. schools provided no instruction on cultural issues (27% versus 8%; p = .04). Few schools taught about the specific cultural issues of the largest minority groups in their geographic areas: only 28% and 26% of U.S. schools taught about African American and Latino issues, respectively, and only two thirds of Canadian schools taught about either Asian or Native Canadian issues. Only 35% of U.S. schools addressed the cultural issues of the largest minority groups in their particular states. CONCLUSIONS: Most U.S. and Canadian medical schools provide inadequate instruction about cultural issues, especially the specific cultural aspects of large minority groups.


Assuntos
Características Culturais , Educação Médica , Faculdades de Medicina , Canadá , Currículo , Estados Unidos
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