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1.
Rofo ; 186(12): 1111-21, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25122171

RESUMO

PURPOSE: To compare the performance of server-based (CSS) versus stand-alone post-processing software (ES) for the evaluation of cardiovascular CT examinations (cvCT) and to determine the crucial steps. MATERIALS AND METHODS: Data of 40 patients (20 patients for coronary artery evaluation and 20 patients prior to transcatheter aortic valve implantation [TAVI]) were evaluated by 5 radiologists with CSS and ES. Data acquisition was performed using a dual-source 128-row CT unit (SOMATOM Definition Flash, Siemens, Erlangen, Germany) and a 64-row CT unit (Brilliance 64, Philips, Hamburg, Germany). The following workflow was evaluated: Data loading, aorta and coronary segmentation, curved multiplanar reconstruction (cMPR) and 3 D volume rendering technique (3D-VRT), measuring of coronary artery stenosis and planimetry of the aortic annulus. The time requirement and subjective quality for the workflow were evaluated. RESULTS: The coronary arteries as well as the TAVI data could be evaluated significantly faster with CSS (5.5  ±  2.9  min and 8.2  ±  4.0  min, respectively) than with ES (13.9  ±  5.2  min and 15.2  ±â€Š 10.9  min, respectively, p ≤  0.01). Segmentation of the aorta (CSS: 1.9 ±  2.0  min, ES: 3.7  ±  3.3  min), generating cMPR of coronaries (CSS: 0.5  ±  0.2  min, ES: 5.1  ±  2.6  min), aorta and iliac vessels (CSS: 0.5  ±  0.4  min and 0.4  ±  0.4  min, respectively, ES: 1.6  ±  0.7  min and 2.8  ±  3  min, respectively) could be performed significantly faster with CSS than with ES with higher quality of cMPR, measuring of coronary stenosis and 3D-VRT (p < 0.05). CONCLUSION: Evaluation of cvCT can be accomplished significantly faster and better with CSS than with ES.  The segmentation remains the most time-consuming workflow step, so optimization of segmentation algorithms could improve performance even further.


Assuntos
Técnicas de Imagem de Sincronização Cardíaca/métodos , Doenças Cardiovasculares/diagnóstico por imagem , Doença das Coronárias/diagnóstico por imagem , Processamento de Imagem Assistida por Computador/métodos , Tomografia Computadorizada Multidetectores/métodos , Software , Substituição da Valva Aórtica Transcateter/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Angioplastia/métodos , Valva Aórtica/diagnóstico por imagem , Feminino , Humanos , Imageamento Tridimensional/métodos , Masculino , Pessoa de Meia-Idade , Estudos de Tempo e Movimento , Fluxo de Trabalho
2.
Radiologe ; 53(10): 886-95, 2013 Oct.
Artigo em Alemão | MEDLINE | ID: mdl-24129987

RESUMO

Percutaneous transcatheter aortic valve implantation (TAVI) is an established alternative to open heart surgery in patients with severe aortic stenosis (AS) unsuitable for conventional aortic valve replacement due to comorbidities with a high perioperative risk or contraindications. Preprocedural imaging plays a major role for adequate determination of indications and prosthesis selection, prosthesis sizing and therefore for a reduction of periprocedural complications. Besides Doppler echocardiography which is mainly used for grading of the severity of aortic valve stenosis and peri-interventional imaging, cardiac computed tomography (CCT) is the imaging modality of choice. The CCT procedure not only allows for reliably assessment and measuring of the complex 3-dimensional geometry of the aortic root but also for the aorta and the peripheral vessels used as potential access paths.


Assuntos
Estenose da Valva Aórtica/diagnóstico por imagem , Estenose da Valva Aórtica/cirurgia , Cateterismo Cardíaco/métodos , Implante de Prótese de Valva Cardíaca/métodos , Radiografia Intervencionista/métodos , Cirurgia Assistida por Computador/métodos , Tomografia Computadorizada por Raios X/métodos , Humanos , Cuidados Pré-Operatórios/métodos , Prognóstico
3.
Rofo ; 184(10): 941-9, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23999787

RESUMO

UNLABELLED: Transcatheter aortic valve implantation (TAVI) is currently considered an acceptable alternative for the treatment of patients with severe aortic stenosis and a high perioperative risk or a contraindication for open surgery. The benefit of TAVI significantly outweighs the risk of the procedure in patients requiring treatment that are not suitable for open surgery, and leads to a lower mortality in the one-year follow-up. The absence of a direct view of the aortic root and valve remains a challenge for the transcatheter approach. While direct inspection of the aortic valve during open surgery allows an adequate prosthesis choice, it is crucial for TAVI to know the individual anatomical details prior to the procedure in order to assure adequate planning of the procedure and proper prosthesis choice and patient selection. Among the imaging modalities available for the evaluation of patients prior to TAVI, computed tomography (CT) plays a central role in patient selection. CT reliably visualizes the dimensions of the aortic root and allows a proper choice of the prosthesis size. The morphology of the access path and relevant comorbidities can be assessed. The present review summarizes the current state of knowledge regarding the value of CT in the evaluation of patients prior to TAVI. KEY POINTS: CT plays a central role in patient selection and planning prior to TAVI. ▶ CT reliably detects the dimensions of the aortic root including the size of the aortic annulus, the degree of valve calcification and the morphology of the access routes. ▶ CT provides a more accurate measurement of the aortic annulus than 2D TEE and CT is the only imaging modality that allows a risk assessment for paravalvular leakages based on the calcification of the aortic valve.


Assuntos
Estenose da Valva Aórtica/diagnóstico por imagem , Estenose da Valva Aórtica/cirurgia , Cateterismo Cardíaco , Implante de Prótese de Valva Cardíaca , Processamento de Imagem Assistida por Computador/métodos , Imageamento Tridimensional/métodos , Tomografia Computadorizada Multidetectores/métodos , Radiografia Intervencionista , Valva Aórtica/diagnóstico por imagem , Valva Aórtica/cirurgia , Calcinose/diagnóstico por imagem , Calcinose/cirurgia , Humanos , Planejamento de Assistência ao Paciente , Seleção de Pacientes , Complicações Pós-Operatórias/diagnóstico por imagem , Complicações Pós-Operatórias/prevenção & controle , Cuidados Pré-Operatórios , Desenho de Prótese , Ajuste de Prótese
4.
Rofo ; 185(10): 941-9, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24490256

RESUMO

UNLABELLED: Transcatheter aortic valve implantation (TAVI) is currently considered an acceptable alternative for the treatment of patients with severe aortic stenosis and a high perioperative risk or a contraindication for open surgery. The benefit of TAVI significantly outweighs the risk of the procedure in patients requiring treatment that are not suitable for open surgery, and leads to a lower mortality in the one-year follow-up. The absence of a direct view of the aortic root and valve remains a challenge for the transcatheter approach. While direct inspection of the aortic valve during open surgery allows an adequate prosthesis choice, it is crucial for TAVI to know the individual anatomical details prior to the procedure in order to assure adequate planning of the procedure and proper prosthesis choice and patient selection. Among the imaging modalities available for the evaluation of patients prior to TAVI, computed tomography (CT) plays a central role in patient selection. CT reliably visualizes the dimensions of the aortic root and allows a proper choice of the prosthesis size. The morphology of the access path and relevant comorbidities can be assessed. The present review summarizes the current state of knowledge regarding the value of CT in the evaluation of patients prior to TAVI. CT plays a central role in patient selection and planning prior to TAVI. CT reliably detects the dimensions of the aortic root including the size of the aortic annulus, the degree of valve calcification and the morphology of the access routes. KEY POINTS: CT plays a central role in patient selection and planning prior to TAVI. CT reliably detects the dimensions of the aortic root including the size of the aortic annulus, the degree of valve calcification and the morphology of the access routes. CT provides a more accurate measurement of the aortic annulus than 2D TEE and CT is the only imaging modality that allows a risk assessment for paravalvular leakages based on the calcification of the aortic valve.


Assuntos
Estenose da Valva Aórtica/diagnóstico por imagem , Estenose da Valva Aórtica/cirurgia , Valva Aórtica/diagnóstico por imagem , Valva Aórtica/cirurgia , Cateterismo Cardíaco/métodos , Implante de Prótese de Valva Cardíaca/métodos , Interpretação de Imagem Assistida por Computador/métodos , Imageamento Tridimensional/métodos , Cuidados Pré-Operatórios/métodos , Tomografia Computadorizada por Raios X/métodos , Idoso , Idoso de 80 Anos ou mais , Aorta Abdominal/diagnóstico por imagem , Calcinose/diagnóstico por imagem , Calcinose/cirurgia , Angiografia Coronária , Estenose Coronária/diagnóstico por imagem , Estenose Coronária/cirurgia , Feminino , Artéria Femoral/diagnóstico por imagem , Humanos , Artéria Ilíaca/diagnóstico por imagem , Complicações Intraoperatórias/diagnóstico por imagem , Complicações Intraoperatórias/cirurgia , Masculino , Seleção de Pacientes , Desenho de Prótese , Ajuste de Prótese/métodos , Seio Aórtico/diagnóstico por imagem , Seio Aórtico/cirurgia
5.
Clin Res Cardiol ; 100(4): 343-50, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21088843

RESUMO

AIM: To evaluate changes of right ventricular (RV) parameters in follow-up examinations after corrected tetralogy of Fallot (TOF) by cardiac magnetic resonance (CMR). METHODS: CMR was performed twice within 4 years in 45 patients using a 1.5 T scanner. RV-volumes and pulmonary-regurgitant-fractions (PRF) were calculated from standard cine-sequences and flow-sensitive gradient-echo images, respectively. Patients were divided into two groups depending on the post-operative (po) interval (group 1 ≤5 years po; group 2 >5 years po) and subgroups depending on type of surgery (transannular vs. non-transannular). Patient groups were compared among each other and differences between 1st and 2nd CMR were assessed. Furthermore, patients were compared with 25 healthy volunteers. RESULTS: Compared with controls RV-size was increased (group 1: p = 0.007; group 2: p < 0.001) and RV function decreased (group 1: p = 0.02; group 2: p < 0.001) in po TOF-patients. PRF was higher in group 2 compared with group 1 (p = 0.04) and significant changes of PRF between 1st and 2nd CMR were found in group 2 (p < 0.01), but not in group 1 (p = 0.29). Compared with the non-transannular subgroup, PRF (p < 0.001) and RV end-diastolic-volume index (RV-EDVI) (p = 0.03) were significantly higher in patients with a transannular patch, EDVI increased between 1st and 2nd CMR. After correction, no significant changes of RV myocardial mass index (RV-MMI) were found. CONCLUSION: After correction of TOF, RV-size, RV-muscle mass (RV-MM) was increased and ejection fraction decreased in "early" follow-up already. Whereas these parameters can remain stable over a long time period, the PRF significantly increased in "late" follow-up dependent on the po interval. Overall, transannular patching went along with higher PRF and bigger RV-size as well as a greater dynamic of these parameters in the time course, which makes this subgroup highly in need of regular follow-up examinations for the optimal timing of re-interventions. In contrast, the increased RV-MM demonstrated no regression po.


Assuntos
Tetralogia de Fallot/fisiopatologia , Tetralogia de Fallot/cirurgia , Função Ventricular Direita/fisiologia , Volume Cardíaco , Estudos de Casos e Controles , Criança , Feminino , Seguimentos , Ventrículos do Coração/fisiopatologia , Humanos , Imageamento por Ressonância Magnética , Masculino , Insuficiência da Valva Pulmonar , Estudos Retrospectivos , Fatores de Tempo , Adulto Jovem
7.
Meat Sci ; 80(2): 505-10, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22063359

RESUMO

In a previous study, we showed that coagulase positive staphylococci, which are often used as indicators for Staphylococcus aureus, are frequently found on pig carcasses. Further characterization of the strains identified only a minor part as S. aureus. Selected non-S. aureus strains were all identified as Staphylococcus hyicus, However, two studies described in this species strains that produce staphylococcal enterotoxins. The aim of the present study was to further characterize such coagulase positive S. hyicus strains isolated from pig carcasses and to assess the results for their food safety relevance. A total of 189 strains from two abattoirs were characterized. Phenotypically, 98.9% showed non-pigmented colonies, 99.5% no haemolysis and 67.7% were egg yolk-positive. DNase activity was found in all but one isolate. Only five of the 189 strains were resistant to the antimicrobials tested. One strain harboured the mecA gene. Exfoliative toxin genes were detected in 31 (16.4%), S. aureus enterotoxin genes in none of the strains. The PFGE genotyping results show only a limited number of clusters. Cluster I included more than 50% of the strains. The fact that similar or closely related PFGE patterns of S. hyicus can be found on carcasses after bleeding in both abattoirs indicates the occurrence of widespread strains in the Swiss pig population. Moreover, the genotyping results revealed a remarkable homogeneity in S. hyicus strains isolated from different process stages in abattoir B, which could indicate a recontamination problem with persisting strains.

8.
Vet Microbiol ; 120(3-4): 292-9, 2007 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-17141430

RESUMO

A total of 142 S. aureus strains isolated from pig carcasses from abattoirs A (n=98) and B (n=44) were characterized by phenotypic and genotypic traits. Phenotypically, 96% showed yellow-pigmented colonies, 63% beta/delta hemolysis, 85% were egg yolk-positive and 99% were positive for clumping factor/protein A. Only 25% of the strains were resistant to the antimicrobials tested (abattoir A: 19%; abattoir B: 39%), especially to penicillin and ampicillin. None of the strains harbored the mecA gene, which is conserved in methicillin-resistant S. aureus. The biofilm associated genes icaA, icaD and bap were present in 100%, 100% and 0% of the strains. Genes for staphylococcal enterotoxin (SE) were detected in 51% (abattoir A) and 14% of the strains (abattoir B). Among strains harboring SE genes (n=56), 63%, 31%, 4% and 2% tested positive for seg/sei, seg, sei and sec, respectively. The amplification of the 3' end of the coagulase gene (coa) yielded amplicons of 400, 436, 602, 682 or 776bp. Coa restriction profile (CRP) analysis using HaeIII resulted in seven patterns (a-d, e1-e3). CRP (c) was detected most frequently at both abattoirs, whereas CRP (a) was restricted to abattoir A and CRP (e3) to abattoir B. In the slaughter process (abattoir B), (i) two CRPs (b and d) were only found before dehairing/singeing, and (ii) four CRPs (c, e1-e3) were identified throughout the process. The genotyping revealed a remarkable homogeneity in S. aureus strains from the two different abattoirs and the slaughter process stages. These results may be explained by the distribution of a limited number of S. aureus genotypes in the pig population. Moreover, as the predominant CRPs (c, e1-e3) persisted throughout the slaughter process in abattoir B, it may be hypothesized that these types are characterized by colonization advantages.


Assuntos
Infecções Estafilocócicas/veterinária , Staphylococcus aureus/genética , Doenças dos Suínos/microbiologia , Animais , Anti-Infecciosos/farmacologia , Moléculas de Adesão Celular/genética , Coagulase/genética , Primers do DNA/química , Enterotoxinas/genética , Genótipo , Testes de Sensibilidade Microbiana/veterinária , Fenótipo , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/classificação , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/isolamento & purificação , Suínos
9.
Eur Heart J ; 25(8): 694-700, 2004 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15084375

RESUMO

AIMS: To investigate the results of endovascular stent-graft placement for the treatment of acute perforating lesions of the descending thoracic aorta. METHODS AND RESULTS: A total of 31 consecutive patients underwent interventional treatment for perforating lesions of the descending aorta. In 21 cases (group A), the aortic perforation was due to rupture of a descending thoracic aneurysm or dissection, whereas 10 patients (group B) were treated for traumatic transection of the descending aorta. A total of 42 endoprostheses were implanted. The implantation procedure was successful in all cases without peri-interventional complications. In one case, implantation of a second endoprosthesis became necessary due to type I endoleak. Overall, the 30-day mortality was 9.7%. As all three deaths occurred in group A, the mortality rate in this group was 14.3% versus 0% in group B. Similarly, postinterventional complications were more prevalent, with 28.6% in group A (renal failure n = 4; stroke n = 2) versus 10.0% in group B (renal failure n = 1). No paraplegia and no further deaths or ruptures occurred during follow-up (mean 17 months). CONCLUSION: Interventional stent-graft placement is an effective treatment option for the emergency repair of descending aortic perforations.


Assuntos
Aneurisma da Aorta Torácica/terapia , Ruptura Aórtica/terapia , Stents , Injúria Renal Aguda/etiologia , Idoso , Idoso de 80 Anos ou mais , Aneurisma da Aorta Torácica/mortalidade , Ruptura Aórtica/mortalidade , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Acidente Vascular Cerebral/etiologia , Tomografia Computadorizada por Raios X/métodos , Resultado do Tratamento
10.
Arch Toxicol ; 75(5): 306-12, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11548124

RESUMO

The DNA repair enzymes O6-methylguanine-DNA methyltransferase (MGMT) and apurinic/apyrimidinic endonuclease (APE, also known as Ref-1) play an important role in cellular defense against the mutagenic and carcinogenic effects of DNA-damaging agents. Cells with low enzyme activity are more sensitive to induced DNA damage and may confer a higher carcinogenic risk to the individuals in question. To study the level of variability of MGMT and APE expression in human, we analyzed in a long-time study MGMT and APE expression in peripheral blood mononuclear cells (PBMC) from healthy individuals. The data revealed high inter- and intraindividual variability of MGMT but not of APE. For MGMT, the interindividual levels ranged from 27 to 204 fmol/10(6) cells (7.6-fold, 40 healthy individuals). The intraindividual variation was determined by measuring MGMT repeatedly over 42 days, and was found to vary from 1.4-fold to 3.5-fold. Averaging over the measurement period, some individuals displayed low MGMT activity compared to others. In contrast, APE expression showed only a 2.9-fold difference between individuals and a 1.2 to 2.3-fold intra-individual long-time variation, and thus was less variable than MGMT. MGMT and APE expression were not correlated. Overall the results showed variable MGMT and rather constant APE levels in PBMC of healthy individuals measured over a long period.


Assuntos
Carbono-Oxigênio Liases/biossíntese , DNA Ligases/metabolismo , Monócitos/enzimologia , O(6)-Metilguanina-DNA Metiltransferase/biossíntese , Adulto , Western Blotting , DNA Liase (Sítios Apurínicos ou Apirimidínicos) , Desoxirribonuclease IV (Fago T4-Induzido) , Feminino , Humanos , Masculino , Fumar/metabolismo , Fatores de Tempo
12.
Arch Toxicol ; 73(1): 15-21, 1999 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-10207610

RESUMO

A recent study reported that exposure of student embalmers in Cincinnati to high concentrations of formaldehyde (2 mg/m3) reduced the activity of the DNA repair protein O6-methylguanine DNA methyltransferase (MGMT). Reduction in a DNA repair enzyme may strongly increase the cancer risk not only with respect to the repair-enzyme causing agent but with respect to all carcinogens causing lesions subject to repair by the enzyme in question. Thus, we examined whether formaldehyde exposure of 57 medical students during their anatomy course at two different Universities in Germany influenced MGMT activity in mononuclear blood cells. Mean formaldehyde exposure of 41 students was 0.2 +/- 0.05 mg/m3 for 6 h per week. MGMT activity was 133.2 +/- 14.9 fmol MGMT/10(6) cells before the beginning of the formaldehyde exposure, 131.1 +/- 15.8 fmol MGMT/10(6) cells after 50 days (P = 0.56) and 128.2 +/- 19.0 fmol MGMT/10(6) cells after 111 days of exposure (P = 0.92). Similarly, no significant influence of formaldehyde exposure was observed, when smoking habits, alcohol consumption, allergic disease and sex of students were considered. In addition no significant difference was obtained in MGMT activity between 16 students with mean formaldehyde exposure of 0.8 +/- 0.6 mg/m3 and students without formaldehyde exposure (n = 51; P = 0.37). In conclusion, exposure of the medical students in western Europe to formaldehyde did not decrease MGMT activity in mononuclear blood cells.


Assuntos
Fixadores/farmacologia , Formaldeído/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , O(6)-Metilguanina-DNA Metiltransferase/efeitos dos fármacos , Estudantes de Medicina , Adulto , Consumo de Bebidas Alcoólicas , Interpretação Estatística de Dados , Exposição Ambiental , Feminino , Humanos , Hipersensibilidade , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/enzimologia , Masculino , O(6)-Metilguanina-DNA Metiltransferase/sangue , Fumar , Fatores de Tempo
13.
Diabetes ; 47(12): 1928-34, 1998 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9836526

RESUMO

The in vivo kinetics of the HDL apolipoproteins (apo) A-I and A-II were studied in six subjects with impaired glucose tolerance (IGT) and six control subjects with normal glucose tolerance (NGT), using a stable isotope approach. During a 12-h primed constant infusion of L-[ring-13C6]-phenylalanine, tracer enrichment was determined in apoA-I and apoA-II from ultracentrifugally isolated HDL. The rates of HDL apoA-I and apoA-II production and catabolism were estimated using a one-compartment model-based analysis. Triglycerides were higher in IGT subjects (1.33 +/- 0.21 vs. 0.84 +/- 0.27 mmol/l, P < 0.05), but were within the normal range. HDL cholesterol and apoA-I levels were significantly lower in subjects with IGT (1.07 +/- 0.15 vs. 1.36 +/- 0.14 mmol/l, P < 0.05; 0.94 +/- 0.10 vs. 1.34 +/- 0.07 g/l, P < 0.01). In IGT subjects, HDL composition was significantly altered, characterized by an increase in HDL triglycerides (4.9 +/- 1.9 vs. 3.2 +/- 1.0%, P < 0.05) and HDL phospholipids (34.7 +/- 2.6 vs. 27.5 +/- 5.8%, P < 0.05) and a decrease in HDL cholesteryl esters (10.1 +/- 2.0 vs. 12.7 +/- 2.9%, P < 0.05) and HDL apoA-I (31.5 +/- 4.4 vs. 43.2 +/- 2.4%, P < 0.05). The mean fractional catabolic rate (FCR) of HDL apoA-I was significantly higher in IGT subjects (0.34 +/- 0.05 vs. 0.26 +/- 0.03 day(-1), P < 0.01), while the HDL apoA-I production rate (PR), as well as the PR and FCR of HDL apoA-II, showed no differences between the two groups. There were significant correlations between HDL apoA-I FCR and the following parameters: HDL apoA-I (r = -0.902, P < 0.001), HDL cholesterol (r = -0.797, P = 0.001), plasma triglycerides (r = 0.743, P < 0.01), HDL triglycerides (r = 0.696, P < 0.01), and cholesterol ester transfer protein activity (r = 0.646, P < 0.01). We observed a strong positive association between increased apoA-I catabolism and insulin (r = 0.765, P < 0.01) and proinsulin (r = 0.797, P < 0.01) concentrations. These data support the hypothesis that the decrease in HDL cholesterol and apoA-I levels in IGT is principally the result of an enhanced apoA-I catabolism. The latter seems to be an early metabolic finding in IGT even when other lipid parameters, especially plasma triglycerides, still appear to be not or only weakly affected.


Assuntos
Apolipoproteína A-I/sangue , Intolerância à Glucose/sangue , Intolerância à Glucose/fisiopatologia , Glicoproteínas , Adulto , Fatores Etários , Apolipoproteína A-II/sangue , Índice de Massa Corporal , Isótopos de Carbono , Proteínas de Transporte/sangue , Proteínas de Transferência de Ésteres de Colesterol , Feminino , Humanos , Cinética , Lipase Lipoproteica/sangue , Lipoproteínas HDL/sangue , Lipoproteínas HDL/química , Masculino , Pessoa de Meia-Idade , Fenilalanina/sangue , Fatores Sexuais , Estatística como Assunto
14.
J Lipid Res ; 37(10): 2074-87, 1996 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-8906585

RESUMO

In familial defective apolipoprotein B-100 (FDB) the presence of a mutant apolipoprotein (apo) B-100 (FDB3500Q/W) in LDL markedly reduces their affinity for the LDL receptor, leading to elevated LDL cholesterol levels. However, the hypercholesterolemia in most FDB patients is relatively mild when compared with, e.g., familial hypercholesterolemia (FH). In order to study mechanisms that may partly alleviate the clinical consequences of FDB, we investigated the in vivo kinetics of apoB-100-containing lipoproteins in five FDB heterozygotes (total cholesterol: 7.84 +/- 1.37 mmol/I; total apoB: 1.68 +/- 0.37 g/l; mean +/- SD) and six normolipidemic controls (4.61 +/- 0.62 mmol/l; 0.98 +/- 0.12 g/l) using a stable isotope approach. During and after a 10-12 h primed, constant infusion of either [13C6]phenylalanine or [2H3]leucine, tracer enrichment was determined in apoB-100 from ultracentrifugally isolated VLDL1 (Sf 60-400), VLDL2 (Sf 20-60), IDL (Sf 12-20), LDL1 (Sf 7-12), and LDL2 (Sf 0-7). The rates of apoB-100 production, catabolism, and transfer were estimated by model-based compartmental analysis. The overall fractional catabolic rate (FCR) of IDL apoB-100 in FDB was substantially increased (2.99 +/- 0.68 pools/day vs. 1.70 +/- 0.23 pools/day in controls, P < 0.01). The fractional rate of apoB-100 transfer from IDL to LDL in FDB was decreased (0.97 +/- 0.13 pools/day vs. 1.24 +/- 0.10 pools/day, P < 0.05). The FCR of LDL apoB-100 in FDB was decreased (0.18 +/- 0.07 pools/day vs. 0.56 +/- 0.05 pools/, P < 0.01). Finally, the input rate of LDL apoB-100 in FDB was markedly decreased (9.45 +/- 2.96 mg/kg day1 vs. 15.54 +/- 1.70 mg/kg day1, P < 0.05). Our data suggest that the relatively small increase of LDL concentrations in FDB is due to an increased clearance of LDL precursor particles via the LDL-receptor and apoE-receptors as well as a decreased conversion of IDL to LDL - two mechanisms that distinguish FDB from FH.


Assuntos
Apolipoproteínas B/genética , Lipoproteínas LDL/sangue , Taxa de Depuração Metabólica , Precursores de Proteínas/sangue , Adulto , Apolipoproteína B-100 , Apolipoproteínas B/sangue , Isótopos de Carbono , Deutério , Feminino , Heterozigoto , Humanos , Cinética , Leucina/sangue , Lipídeos/sangue , Lipoproteínas/sangue , Masculino , Pessoa de Meia-Idade , Mutação , Fenilalanina/sangue
15.
Biochim Biophys Acta ; 1254(1): 77-88, 1995 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-7811751

RESUMO

A very short run time and small sample volumes in the separation of lipoproteins by preparative ultracentrifugation are needed for several investigations. Recently, a very fast sequential separation method was described that needs only 100 min for one run in a centrifugal field of 625,000 x g. We studied the influence of centrifugal fields of this dimension on lipoprotein separation and lipoprotein particle integrity using a Beckman Optima TLX ultracentrifuge with a TLA-120.2 rotor. Rotor speed (120/90/60/30.10(3) rev./min) and run time (100 min/3 h/6.7 h/27 h) were selected in such a way that the product of centrifugal field and run time remained constant. The first conditions correspond to the very fast ultracentrifugation (VFU) procedure with a centrifugal field of 625,000 x g. Thirty different plasma samples covering a wide range of lipid and protein concentrations were separated in the course of two centrifugal runs at densities of 1.006 and 1.063 kg/l which yielded very-low-density lipoproteins (VLDL), low-density lipoproteins (LDL), and the subnatant of low-density lipoproteins, including high-density lipoproteins (HDL) and concomitant sedimented plasma proteins. The major lipid components of the lipoproteins, triacylglycerols, free and esterified cholesterol, phospholipids and the apolipoproteins B and A-I, were estimated considering the masses of the tube contents after a slicing procedure. Measurements of lipids and proteins showed a very good recovery of better than 94% and 91%, respectively, and precision-within-series (coefficient of variation) of better than 4.2% and 6.5%, respectively. The effects of the rotor speed on the lipoprotein structure appeared to be weak. With increasing rotor speed, VLDL and LDL lipid constituents principally tended to decrease, whereas they increased in the subnatant of the LDL-run. The mean lipoprotein mass composition, considering the mass percentage of each measured particle constituent, did not show significant alterations. Total protein decreased in VLDL and in LDL and increased in the subnatant of the LDL-run. As checked by an enzyme-linked immunosorbent assay (ELISA) and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), the protein effects were due to nearly complete disappearance of contaminating plasma proteins, especially albumin as the major contamination of VLDL and LDL. The apolipoproteins (apo) B-100, A-I, E and C-I to C-III remained nearly unaffected. The main advantages of VFU were the very short run time (cumulative flotation time is 3.4 h) and the elemination of albumin without repeated runs. The procedure was suitable for the assessment of lipid and protein constituents in lipoproteins from very small plasma samples (500 microliters).


Assuntos
Lipoproteínas/isolamento & purificação , Ultracentrifugação/métodos , Adulto , Proteínas Sanguíneas/análise , Eletroforese em Gel de Poliacrilamida , Humanos , Hiperlipidemias/sangue , Immunoblotting , Lipoproteínas/sangue , Lipoproteínas/química , Masculino , Pessoa de Meia-Idade , Albumina Sérica/análise
16.
Clin Chim Acta ; 224(1): 21-32, 1994 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-8174275

RESUMO

A short run-time in separation of lipoproteins by preparative ultracentrifugation is desirable for several reasons. Recently, a method was described that needs only 100 min for one run in a centrifugal field of 625,000 x g. It is assumed that lipoprotein separation depends on the rotor speed but this has not been systematically studied in centrifugal fields of this order. We performed such a study. Rotor speeds of 120, 90, 60 and 30 x 10(3) rev./min and run-times of 100 min, 3 h, 6.7 h and 27 h were selected in such a way that the product of centrifugal field and run-time remained constant. The first conditions correspond to the 'very fast ultracentrifugation' (VFU) procedure with a centrifugal field of 625,000 x g. The Optima tabletop ultracentrifuge, the rotor TLA-120.2 and thick wall open tubes for 1 ml were used. Thirteen different plasma samples covering a wide range of cholesterol and triglyceride concentrations were separated into VLDL, LDL and HDL in the course of two centrifugal runs at densities of 1.006 and 1.063. The constituents of the lipoproteins were calculated considering the mass of the tube contents after slicing. Recoveries of cholesterol, triglycerides and protein were 97%, 98% and 90%, respectively. The influence of the rotor speed on the apparent composition of the lipoproteins was small. With increasing rotor speed, VLDL-cholesterol became higher (by 14%, P < 0.001), VLDL-triglyceride became lower (by 6%, P < 0.012), LDL-cholesterol became lower (by 9%, P < 0.000). The effects on LDL-triglyceride and on HDL-cholesterol and HDL-triglyceride, did not reach statistical significance. Protein in VLDL and in LDL decreased and increased in 'HDL' (the subnatant of the LDL run). As checked by SDS-PAGE the protein effects were due to complete disappearance of albumin from VLDL and LDL while the apolipoproteins B-100, E and C-I to C-III remained unaffected. It is concluded that the main advantages of VFU are the short run-time and the disappearance of albumin from VLDL and LDL. The other compositional changes need to be further investigated.


Assuntos
Lipoproteínas/sangue , Apolipoproteínas/metabolismo , Colesterol/sangue , Eletroforese em Gel de Poliacrilamida , Humanos , Lipoproteínas/química , Lipoproteínas/isolamento & purificação , Lipoproteínas HDL/sangue , Lipoproteínas LDL/sangue , Lipoproteínas VLDL/sangue , Triglicerídeos/sangue , Ultracentrifugação
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