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1.
Mar Pollut Bull ; 110(2): 701-17, 2016 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-27179997

RESUMO

Substrate cover, water quality parameters and assemblages of corals, fishes, sponges, echinoderms, ascidians, molluscs, benthic foraminifera and macroalgae were sampled across a pronounced environmental gradient in the Jakarta Bay-Thousand Islands reef complex. Inshore sites mainly consisted of sand, rubble and turf algae with elevated temperature, dissolved oxygen, pH and chlorophyll concentrations and depauperate assemblages of all taxa. Live coral cover was very low inshore and mainly consisted of sparse massive coral heads and a few encrusting species. Faunal assemblages were more speciose and compositionally distinct mid- and offshore compared to inshore. There were, however, small-scale differences among taxa. Certain midshore sites, for example, housed assemblages resembling those typical of the inshore environment but this differed depending on the taxon. Substrate, water quality and spatial variables together explained from 31% (molluscs) to 72% (foraminifera) of the variation in composition. In general, satellite-derived parameters outperformed locally measured parameters.


Assuntos
Antozoários/crescimento & desenvolvimento , Organismos Aquáticos/classificação , Baías/química , Monitoramento Ambiental/métodos , Água do Mar/química , Animais , Antozoários/efeitos dos fármacos , Organismos Aquáticos/efeitos dos fármacos , Organismos Aquáticos/crescimento & desenvolvimento , Biodiversidade , Recifes de Corais , Equinodermos/crescimento & desenvolvimento , Ecossistema , Peixes/crescimento & desenvolvimento , Foraminíferos/crescimento & desenvolvimento , Indonésia , Ilhas , Moluscos/crescimento & desenvolvimento , Poríferos/crescimento & desenvolvimento , Alga Marinha/crescimento & desenvolvimento , Urbanização , Urocordados/crescimento & desenvolvimento , Qualidade da Água
2.
Med Image Anal ; 16(1): 127-39, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21719343

RESUMO

Endovascular aortic replacement (EVAR) is an established technique, which uses stent grafts to treat aortic aneurysms in patients at risk of aneurysm rupture. Late stent graft failure is a serious complication in endovascular repair of aortic aneurysms. Better understanding of the motion characteristics of stent grafts will be beneficial for designing future devices. In addition, analysis of stent graft movement in individual patients in vivo can be valuable for predicting stent graft failure in these patients. To be able to gather information on stent graft motion in a quick and robust fashion, we propose an automatic method to segment stent grafts from CT data, consisting of three steps: the detection of seed points, finding the connections between these points to produce a graph, and graph processing to obtain the final geometric model in the form of an undirected graph. Using annotated reference data, the method was optimized and its accuracy was evaluated. The experiments were performed using data containing the AneuRx and Zenith stent grafts. The algorithm is robust for noise and small variations in the used parameter values, does not require much memory according to modern standards, and is fast enough to be used in a clinical setting (65 and 30s for the two stent types, respectively). Further, it is shown that the resulting graphs have a 95% (AneuRx) and 92% (Zenith) correspondence with the annotated data. The geometric model produced by the algorithm allows incorporation of high level information and material properties. This enables us to study the in vivo motions and forces that act on the frame of the stent. We believe that such studies will provide new insights into the behavior of the stent graft in vivo, enables the detection and prediction of stent failure in individual patients, and can help in designing better stent grafts in the future.


Assuntos
Aneurisma Aórtico/diagnóstico por imagem , Aneurisma Aórtico/cirurgia , Aortografia/métodos , Prótese Vascular , Reconhecimento Automatizado de Padrão/métodos , Stents , Tomografia Computadorizada por Raios X/métodos , Algoritmos , Angiografia Coronária/métodos , Humanos , Intensificação de Imagem Radiográfica/métodos , Interpretação de Imagem Radiográfica Assistida por Computador/métodos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
3.
Methods Mol Biol ; 771: 337-51, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21874487

RESUMO

Magnetic resonance spectroscopic imaging (MRSI) of the mouse brain reveals a wealth of metabolic information, not only from a single region of interest (single voxel), but spatially mapped over potentially the entire brain. However, MRSI requires challenging methods before the data can be obtained accurately. When applied in vivo, MRSI is generally combined with volume-selective spin perturbation to exclude artifact originating from outside the volume of interest. To obtain good magnetic field (B (0)) uniformity at this volume, accurate B (0) shimming is required. Finally, the immensely large signals originating from water spins need to be suppressed to prevent sidebands that contaminate the spectra, or even saturate the dynamic range of the MR receiver. This chapter describes solutions for these challenges and ends with a rationale between single-voxel MRS versus MRSI.


Assuntos
Encéfalo/patologia , Imageamento por Ressonância Magnética/métodos , Animais , Camundongos , Ratos
4.
Cardiovasc Intervent Radiol ; 34(1): 180-3, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20668850

RESUMO

PURPOSE: To evaluate the accuracy and technical success of positioning a radiofrequency ablation (RFA) electrode in osteoid osteomas by use of a new real-time needle guidance technology combining cone-beam computed tomography (CT) and fluoroscopy. MATERIALS AND METHODS: Percutaneous RFA of osteoid osteomas was performed in five patients (median age 18 years), under general anesthesia, with the use of cone-beam CT and fluoroscopic guidance for electrode positioning. The outcome parameters were technical success, meaning correct needle placement in the nidus; accuracy defined as the deviation (in mm) from the center of the nidus; and clinical outcome at follow-up. RESULTS: In all five cases, positioning was possible within 3 mm of the determined target location (median nidus size 6.8 mm; range 5-10.2 mm). All procedures were technically successful. All patients were free of pain at clinical follow-up. No complications were observed. CONCLUSION: Real-time fluoroscopy needle guidance based on cone-beam CT is a useful tool to accurately position radiofrequency needles for minimally invasive treatment of osteoid osteomas.


Assuntos
Neoplasias Ósseas/cirurgia , Ablação por Cateter/instrumentação , Osteoma Osteoide/cirurgia , Adolescente , Neoplasias Ósseas/diagnóstico por imagem , Pré-Escolar , Tomografia Computadorizada de Feixe Cônico , Feminino , Fluoroscopia , Humanos , Masculino , Pessoa de Meia-Idade , Agulhas , Osteoma Osteoide/diagnóstico por imagem , Radiografia Intervencionista , Resultado do Tratamento
5.
Med Phys ; 37(9): 4744-50, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20964192

RESUMO

PURPOSE: To introduce a new in-house developed pneumatically controlled magnetic field compatible manipulator as an aid to perform magnetic resonance (MR)-guided biopsies of the prostate. METHODS: A pneumatic controlled manipulator with five degrees of freedom constructed of plastic to achieve magnetic field compatibility was developed to guide biopsies. A risk analysis, mechanical tests, and RF safety tests with respect to needle tip heating were performed to assure future patient safety and to meet standard safety requirements for the use in a medical environment. The accuracy of needle positioning with the needle guide manipulator to sample a predefined target was measured in agar phantoms on a 3 T whole body MR system. The in-plane error was used to evaluate the accuracy, which is defined as the orthogonal distance between target and biopsy needle. The time for each step in the biopsy procedure was recorded to evaluate the procedure time. The influence of the insertion angle with respect to the static field of the MR scanner on the needle artifact was investigated. RESULTS: The risk analyses met patient safety requirements. No RF induced local heating around the needle tip was observed. The average in-plane error in 19 measurements was 3.0 mm (range 0-5.6 mm). The average time needed for manipulation to place the needle guide in the desired position was 5 min (range 3-8 min). Total procedure time was 30 min. The needle artifact size increases with the insertion angle with respect to the static field of the MR scanner. CONCLUSIONS: The new MR compatible manipulator can be used safely for patient care. It showed a high accuracy and short total procedure time, demonstrating great potential to improve the transrectal prostate biopsy procedure.


Assuntos
Biópsia/efeitos adversos , Biópsia/métodos , Próstata/patologia , Reto , Robótica/métodos , Segurança , Humanos , Imageamento por Ressonância Magnética , Masculino , Imagens de Fantasmas , Cirurgia Assistida por Computador
6.
Med Phys ; 36(10): 4616-24, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19928093

RESUMO

PURPOSE: ECG-gated CT enables the visualization of motions caused by the beating of the heart. Although ECG gating is frequently used in cardiac CT imaging, this technique is also very promising for evaluating vessel wall motion of the aortic artery and the motions of (stent grafts inside) abdominal aortic aneurysms (AAA). Late stent graft failure is a serious complication in endovascular repair of aortic aneurysms. Better understanding of the motion characteristics of stent grafts will be beneficial for designing future devices. In addition, these data can be valuable in predicting stent graft failure in patients. To be able to reliably quantify the motion, however, it is of importance to know the performance and limitations of ECG gating, especially when the motions are small, as is the case in AAA. Since the details of the reconstruction algorithms are proprietary information on the CT manufacturers and not in the public domain, empirical experiments are required. The goal of this study is to investigate as to what extent the motions in AAA can be measured using ECG-gated CT. The authors quantitatively investigate four aspects of motion in ECG-gated CT: The detectability of the motion of objects at different amplitudes and different periodic motions, the temporal resolution, and the volume gaps that occur as a function of heart rate. METHODS: They designed an experiment on a standard static phantom to empirically determine temporal resolution. To investigate motion amplitude and frequency, as well as patient heart rate, they designed dynamic experiments in which a home-made phantom driven by a motion unit moves in a predetermined pattern. RESULTS: The duration of each ECG-gated phase was found to be 185 ms, which corresponds to half of the rotation time and is thus in accordance with half scan reconstruction applied by the scanner. By using subpixel localization, motions become detectable from amplitudes of as small as 0.4 mm in the x direction and 0.7 mm in the z direction. With the rotation time used in this study, motions up to 2.7 Hz can be reliably detected. The reconstruction algorithm fills volume gaps with noisy data using interpolation, but objects within these gaps remain hidden. CONCLUSIONS: This study gives insight into the possibilities and limitations for measuring small motions using ECG-gated CT. Application of the experimental method is not restricted to the CT scanner of a single manufacturer. From the results, they conclude that ECG-gated CTA is a suitable technique for studying the expected motions of the stent graft and vessel wall in AAA.


Assuntos
Angiografia/métodos , Aneurisma da Aorta Abdominal/diagnóstico por imagem , Artefatos , Técnicas de Imagem de Sincronização Cardíaca/métodos , Reconhecimento Automatizado de Padrão/métodos , Interpretação de Imagem Radiográfica Assistida por Computador/métodos , Tomografia Computadorizada por Raios X/métodos , Algoritmos , Humanos , Movimento (Física) , Imagens de Fantasmas , Intensificação de Imagem Radiográfica/métodos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Tomografia Computadorizada por Raios X/instrumentação
7.
J Endovasc Ther ; 16(5): 546-51, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19842723

RESUMO

PURPOSE: To quantify dynamic changes in aortoiliac dimensions using dynamic electrocardiographically (ECG)-gated computed tomographic angiography (CTA) and to investigate any potential impact on preoperative endograft sizing in relation to observer variability. METHODS: Dynamic ECG-gated CTA was performed in 18 patients with abdominal aortic aneurysms. Postprocessing resulted in 11 datasets per patient: 1 static CTA and 10 dynamic CTA series. Vessel diameter, length, and angulation were measured for all phases of the cardiac cycle. The differences between diastolic and systolic aneurysm dimensions were analyzed for significance using paired t tests. To assess intraobserver variability, 20 randomly selected datasets were analyzed twice. Intraobserver repeatability coefficients (RC) were calculated using Bland-Altman analysis. RESULTS: Mean aortic diameter at the proximal neck was 21.4+/-3.0 mm at diastole and 23.2+/-2.9 mm at systole, a mean increase of 1.8+/-0.4 mm (8.5%, p<0.01). The RC for the aortic diameter at the level of the proximal aneurysm neck was 1.9 mm (8.9%). At the distal sealing zones, the mean increase in diameter was 1.7+/-0.3 mm (14.1%, p<0.01) for the right and 1.8+/-0.5 mm (14.2%, p<0.01) for the left common iliac artery (CIA). At both distal sealing zones, the mean increase in CIA diameter exceeded the RC (10.0% for the right CIA and 12.6% for the left CIA). CONCLUSION: The observed changes in aneurysm dimension during the cardiac cycle are small and in the range of intraobserver variability, so dynamic changes in proximal aneurysm neck diameter and aneurysm length likely have little impact on preoperative endograft selection. However, changes in diameter at the distal sealing zones may be relevant to sizing, so distal oversizing of up to 20% should be considered to prevent distal type I endoleak.


Assuntos
Aneurisma da Aorta Abdominal/diagnóstico por imagem , Aneurisma da Aorta Abdominal/cirurgia , Aortografia/métodos , Implante de Prótese Vascular/instrumentação , Prótese Vascular , Stents , Tomografia Computadorizada por Raios X , Eletrocardiografia , Humanos , Variações Dependentes do Observador , Seleção de Pacientes , Valor Preditivo dos Testes , Desenho de Prótese , Interpretação de Imagem Radiográfica Assistida por Computador , Reprodutibilidade dos Testes , Fatores de Tempo
8.
Science ; 321(5889): 654-7, 2008 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-18669854

RESUMO

Hotspots of high species diversity are a prominent feature of modern global biodiversity patterns. Fossil and molecular evidence is starting to reveal the history of these hotspots. There have been at least three marine biodiversity hotspots during the past 50 million years. They have moved across almost half the globe, with their timing and locations coinciding with major tectonic events. The birth and death of successive hotspots highlights the link between environmental change and biodiversity patterns. The antiquity of the taxa in the modern Indo-Australian Archipelago hotspot emphasizes the role of pre-Pleistocene events in shaping modern diversity patterns.


Assuntos
Biodiversidade , Fósseis , Biologia Marinha , Água do Mar , Animais , Antozoários/classificação , Clima , Ecossistema , Peixes/classificação , Fenômenos Geológicos , Geologia , Moluscos/classificação , Filogenia , Rhizophoraceae/classificação , Tempo
10.
Med Phys ; 34(9): 3510-9, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17926954

RESUMO

The objective of our study was the determination of the influence of the sequential and spiral acquisition modes on the concordance and deviation of the calcium score on 64-slice multi-detector computed tomography (MDCT) scanners in comparison to electron beam tomography (EBT) as the gold standard. Our methods and materials were an anthropomorphic cardio CT phantom with different calcium inserts scanned in sequential and spiral acquisition modes on three identical 64-slice MDCT scanners of manufacturer A and on three identical 64-slice MDCT scanners of manufacturer B and on an EBT system. Every scan was repeated 30 times with and 15 times without a small random variation in the phantom position for both sequential and spiral modes. Significant differences were observed between EBT and 64-slice MDCT data for all inserts, both acquisition modes, and both manufacturers of MDCT systems. High regression coefficients (0.90-0.98) were found between the EBT and 64-slice MDCT data for both scoring methods and both systems with high correlation coefficients (R2>0.94). System A showed more significant differences between spiral and sequential mode than system B. Almost no differences were observed in scanners of the same manufacturer for the Agatston score and no differences for the Volume score. The deviations of the Agatston and Volume scores showed regression dependencies approximately equal to the square root of the absolute score. The Agatston and Volume scores obtained with 64-slice MDCT imaging are highly correlated with EBT-obtained scores but are significantly underestimated (-10% to -2%) for both sequential and spiral acquisition modes. System B is more independent of acquisition mode to calcium score than system A. The Volume score shows no intramanufacturer dependency and its use is advocated versus the Agatston score. Using the same cut points for MDCT-based calcium scores as for EBT-based calcium scores can result in classifying individuals into a too low risk category. System information and scanprotocol is therefore needed for every calcium score procedure to ensure a correct clinical interpretation of the obtained calcium score results.


Assuntos
Calcinose/diagnóstico por imagem , Imagens de Fantasmas , Tomografia Computadorizada por Raios X/métodos , Doença da Artéria Coronariana/diagnóstico por imagem , Tomógrafos Computadorizados
11.
NMR Biomed ; 20(4): 448-67, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17274105

RESUMO

Studies of transgenic mice provide powerful means to investigate the in vivo biological significance of gene products. Mice with an under- or overexpression of enzymes involved in high-energy phosphoryl transfer (approximately P) are particulary attractive for in vivo MR spectroscopy studies as the substrates of these enzymes are metabolites that are visible in MR spectra. This review provides a brief overview of the strategies used for generation and study of genetically altered mice and introduces the reader to some practical aspects of in vivo MRS studies on mice. The major part of the paper reviews results of in vivo MRS studies on transgenic mice with alterations in the expression of enzymes involved in approximately P metabolism, such as creatine kinase, adenylate kinase and guanidinoacetate methyl transferase. The particular metabolic consequences of these enzyme deficiencies in skeletal muscle, brain, heart and liver are addressed. Additionally, the use of approximately P systems as markers of gene expression by MRS, such as after viral transduction of genes, is described. Finally, a compilation of tissue levels of metabolites in skeletal muscle, heart and brain of wild-type and transgenic mice, as determined by in vivo MRS, is given. During the last decade, transgenic MRS studies have contributed significantly to our understanding of the physiological role of phosphotransfer enzymes, and to the view that these enzymes together build a much larger metabolic energy network that is highly versatile and can dynamically adapt to intrinsic genotoxic and extrinsic physiological challenges.


Assuntos
Encéfalo/metabolismo , Creatina Quinase/metabolismo , Creatina/metabolismo , Espectroscopia de Ressonância Magnética/métodos , Músculo Esquelético/metabolismo , Compostos de Fósforo/metabolismo , Animais , Creatina Quinase/genética , Metabolismo Energético/fisiologia , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Fosforilação
12.
Subcell Biochem ; 46: 119-48, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-18652075

RESUMO

Mice with an under- or over-expression of enzymes catalyzing phosphoryl transfer in high-energy supplying reactions are particulary attractive for in vivo magnetic resonance spectroscopy (MRS) studies as substrates of these enzymes are visible in MR spectra. This chapter reviews results of in vivo MRS studies on transgenic mice with alterations in the expression of the enzymes creatine kinase and guanidinoacetate methyltransferase. The particular metabolic consequences of these enzyme deficiencies in skeletal muscle, brain, heart and liver are addressed. An overview is given of metabolite levels determined by in vivo MRS in skeletal muscle and brain of wild-type and transgenic mice. MRS studies on mice lacking guanidinoacetate methyltransferase have demonstrated metabolic changes comparable to those found in the deficiency of this enzyme in humans, which are (partly) reversible upon creatine feeding. Apart from being a model for a creatine deficiency syndrome, these mice are also of interest to study fundamental aspects of the biological role of creatine. MRS studies on transgenic mice lacking creatine kinase isoenzymes have contributed significantly to the view that the creatine kinase reaction together with other enzymatic steps involved in high-energy phosphate transfer builds a large metabolic energy network, which is highly versatile and can dynamically adapt to genotoxic or physiological challenges.


Assuntos
Creatina Quinase/biossíntese , Creatinina/metabolismo , Regulação Enzimológica da Expressão Gênica , Guanidinoacetato N-Metiltransferase/biossíntese , Espectroscopia de Ressonância Magnética , Camundongos Transgênicos , Animais , Creatina Quinase/deficiência , Creatina Quinase/genética , Dano ao DNA/genética , Metabolismo Energético , Regulação Enzimológica da Expressão Gênica/genética , Guanidinoacetato N-Metiltransferase/deficiência , Guanidinoacetato N-Metiltransferase/genética , Humanos , Isoenzimas/biossíntese , Isoenzimas/deficiência , Isoenzimas/genética , Erros Inatos do Metabolismo/enzimologia , Erros Inatos do Metabolismo/genética , Camundongos , Especificidade de Órgãos/genética , Fosfatos/metabolismo
13.
Magn Reson Med ; 55(2): 271-8, 2006 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-16372278

RESUMO

A new coil design for sensitivity-enhanced 13C MR spectroscopy (MRS) of the human brain is presented. The design includes a quadrature transmit/receive head coil optimized for 13C MR sensitivity. Loss-less blocking circuits inside the coil conductors allow this coil to be used inside a homogeneous circularly polarized 1H B1 field for 1H decoupled 13C MRS. A quadrature 1H birdcage coil optimized for minimal local RF heating makes broadband 1H decoupling in the entire human brain possible at 3 Tesla while remaining well within international safety guidelines for RF absorption. Apart from a substantial increase in sensitivity compared to conventional small linear coils, the quadrature 13C coil combined with the quadrature 1H birdcage coil allows efficient cross polarization (CP) in the brain, resulting in an additional 3.5-fold sensitivity improvement compared to direct 13C measurements without nuclear Overhauser enhancement (NOE) or polarization transfer. Combined with the gain in power efficiency, this setup allows broadband 1H to 13C CP over large areas of the brain. Clear 13C resonances from glutamate (Glu), glutamine (Gln), aspartate (Asp), lactate (Lac), and gamma-aminobutyrate (GABA) carbon spins in the human brain demonstrate the quality of 13C MR spectra obtained in vivo with this coil setup.


Assuntos
Química Encefálica , Espectroscopia de Ressonância Magnética/instrumentação , Ácido Aspártico/química , Isótopos de Carbono , Ácido Glutâmico/química , Glutamina/química , Humanos , Aumento da Imagem/métodos , Lactatos/química , Sensibilidade e Especificidade , Processamento de Sinais Assistido por Computador , Ácido gama-Aminobutírico/química
14.
NMR Biomed ; 18(6): 345-51, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16007723

RESUMO

The injection of 1.0 M glutamate into the masseter (jaw-closer) muscle results in a short period of muscle pain (5-10 min) and a prolonged period of mechanical sensitization (> 30 min). It is unclear, however, whether there is a temporal relationship between intramuscular glutamate concentration and either muscle pain or mechanical sensitization. In the present study, (1)H MRS and electrophysiological recording of masticatory muscle nerve fibers were performed in order to monitor glutamate clearance and nerve fiber activity, respectively, after injection of glutamate into rat masticatory muscles. Glutamate signal amplitude was found to decay rapidly (half-life t 1/2 = 108 +/- 42 s), and became indistinguishable from the baseline 10 min after the injection. Glutamate-evoked nerve fiber activity was also found to decay rapidly (t 1/2 = 76 +/- 28 s). These results suggest that glutamate clearance correlates well with the time course of glutamate-evoked muscle pain fiber discharge.


Assuntos
Modelos Animais de Doenças , Ácido Glutâmico/farmacocinética , Espectroscopia de Ressonância Magnética/métodos , Músculos da Mastigação/metabolismo , Síndrome da Disfunção da Articulação Temporomandibular/metabolismo , Animais , Ácido Glutâmico/administração & dosagem , Injeções Intramusculares , Masculino , Músculos da Mastigação/efeitos dos fármacos , Músculos da Mastigação/inervação , Taxa de Depuração Metabólica , Ratos , Ratos Sprague-Dawley , Síndrome da Disfunção da Articulação Temporomandibular/induzido quimicamente , Fibras Aferentes Viscerais/efeitos dos fármacos , Fibras Aferentes Viscerais/metabolismo
15.
J Neurochem ; 90(6): 1321-30, 2004 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-15341516

RESUMO

Creatine kinase (CK)-catalysed ATP-phosphocreatine (PCr) exchange is considered to play a key role in energy homeostasis of the brain. This study assessed the metabolic and anatomical consequences of partial or complete depletion of this system in transgenic mice without cytosolic B-CK (B-CK-/-), mitochondrial ubiquitous CK (UbCKmit-/-), or both isoenzymes (CK -/-), using non-invasive quantitative magnetic resonance (MR) imaging and spectroscopy. MR imaging revealed an increase in ventricle size in a subset of B-CK-/- mice, but not in animals with UbCKmit or compound CK mutations. Mice lacking single CK isoenzymes had normal levels of high-energy metabolites and tissue pH. In the brains of CK double knockouts pH and ATP and Pi levels were also normal, even though PCr had become completely undetectable. Moreover, a 20-30% decrease was observed in the level of total creatine and a similar increase in the level of neuronal N-acetyl-aspartate compounds. Although CKs themselves are not evenly distributed throughout the CNS, these alterations were uniform and concordant across different brain regions. Changes in myo-inositol and glutamate peaks did appear to be mutation type and brain area specific. Our results challenge current models for the biological significance of the PCr-CK energy system and suggest a multifaceted role for creatine in the brain.


Assuntos
Química Encefálica/fisiologia , Encéfalo/metabolismo , Creatina Quinase/deficiência , Isoenzimas/deficiência , Imageamento por Ressonância Magnética/métodos , Ressonância Magnética Nuclear Biomolecular/métodos , Animais , Encéfalo/patologia , Creatina/metabolismo , Creatina Quinase/genética , Creatina Quinase/fisiologia , Creatina Quinase Forma BB , Creatina Quinase Mitocondrial , Isoenzimas/genética , Isoenzimas/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Isótopos de Fósforo/metabolismo , Trítio/metabolismo
16.
J Physiol ; 560(Pt 1): 219-29, 2004 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-15284341

RESUMO

The effects of creatine (Cr) absence in skeletal muscle caused by a deletion of guanidinoacetate methyltransferase (GAMT) were studied in a knockout mouse model by in vivo (31)P magnetic resonance (MR) spectroscopy. (31)P MR spectra of hindleg muscle of GAMT-deficient (GAMT-/-) mice showed no phosphocreatine (PCr) signal and instead showed the signal for phosphorylated guanidinoacetate (PGua), the immediate precursor of Cr, which is not normally present. Tissue pH did not differ between wild-type (WT) and GAMT-/- mice, while relative inorganic phosphate (P(i)) levels were increased in the latter. During ischaemia, PGua was metabolically active in GAMT-/- mice and decreased at a rate comparable to the decrease of PCr in WT mice. However, the recovery rate of PGua in GAMT-/- mice after ischaemia was reduced compared to PCr in WT mice. Saturation transfer measurements revealed no detectable flux from PGua to gamma-ATP, indicating severely reduced enzyme kinetics. Supplementation of Cr resulted in a rapid increase in PCr signal intensity until only this resonance was visible, along with a reduction in relative P(i) values. However, the PGua recovery rate after ischaemia did not change. Our results show that despite the absence of Cr, GAMT-/- mice can cope with mild ischaemic stress by using PGua for high energy phosphoryl transfer. The reduced affinity of creatine kinase (CK) for (P)Gua only becomes apparent during recovery from ischaemia. It is argued that absence of Cr causes the higher relative P(i) concentration also observed in animals lacking muscle CK, indicating an important role of the CK system in P(i) homeostasis.


Assuntos
Glicina/análogos & derivados , Glicina/metabolismo , Metiltransferases/genética , Músculo Esquelético/metabolismo , Fosfocreatina/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Creatina/farmacologia , Guanidinoacetato N-Metiltransferase , Homeostase/fisiologia , Isquemia/metabolismo , Metiltransferases/metabolismo , Camundongos , Camundongos Knockout , Fenótipo , Fosforilação , Descanso/fisiologia
18.
Hum Mol Genet ; 13(9): 905-21, 2004 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-15028668

RESUMO

We generated a knockout mouse model for guanidinoacetate N-methyltransferase (GAMT) deficiency (MIM 601240), the first discovered human creatine deficiency syndrome, by gene targeting in embryonic stem cells. Disruption of the open reading frame of the murine GAMT gene in the first exon resulted in the elimination of 210 of the 237 amino acids present in mGAMT. The creation of an mGAMT null allele was verified at the genetic, RNA and protein levels. GAMT knockout mice have markedly increased guanidinoacetate (GAA) and reduced creatine and creatinine levels in brain, serum and urine, which are key findings in human GAMT patients. In vivo (31)P magnetic resonance spectroscopy showed high levels of PGAA and reduced levels of creatine phosphate in heart, skeletal muscle and brain. These biochemical alterations were comparable to those found in human GAMT patients and can be attributed to the very similar GAMT expression patterns found by us in human and mouse tissues. We provide evidence that GAMT deficiency in mice causes biochemical adaptations in brain and skeletal muscle. It is associated with increased neonatal mortality, muscular hypotonia, decreased male fertility and a non-leptin-mediated life-long reduction in body weight due to reduced body fat mass. Therefore, GAMT knockout mice are a valuable creatine deficiency model for studying the effects of high-energy phosphate depletion in brain, heart, skeletal muscle and other organs.


Assuntos
Peso Corporal/fisiologia , Deficiências Nutricionais/metabolismo , Guanidinas/metabolismo , Metiltransferases/deficiência , Metiltransferases/metabolismo , Animais , Animais Recém-Nascidos , Composição Corporal/genética , Encéfalo/metabolismo , Encéfalo/patologia , Encéfalo/fisiopatologia , Deficiências Nutricionais/genética , Modelos Animais de Doenças , Fertilidade/genética , Guanidinoacetato N-Metiltransferase , Homeostase/fisiologia , Humanos , Técnicas In Vitro , Infertilidade Masculina/genética , Espectroscopia de Ressonância Magnética/métodos , Masculino , Metiltransferases/genética , Camundongos , Camundongos Mutantes , Hipotonia Muscular/genética , Músculo Esquelético/metabolismo , Músculo Esquelético/patologia , Músculo Esquelético/fisiopatologia , Contração Miocárdica/genética
19.
Magn Reson Med ; 50(5): 936-43, 2003 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-14587004

RESUMO

As a model for guanidinoacetate methyltransferase (GAMT) deficiency in humans, a gene knockout mouse model was generated. Here we report on several metabolic abnormalities in these mice, observed by in vivo and in vitro MR spectroscopy. In (1)H MR spectra of brain and hindleg muscle a clearly reduced signal of creatine (Cr) was observed in GAMT-deficient (GAMT-/-) animals. Analysis of the (1)H MR spectra of GAMT-/- brain indicated little or no increase of a signal for guanidinoacetate (Gua). In proton MR spectra of muscle, a broad signal of low intensity was observed for Gua. However, substantial Gua accumulation in intact muscle tissue was unequivocally confirmed in high-resolution magic angle spinning spectra, in which the Gua signal was resolved as one clear sharp singlet. In (31)P MR analysis of brain and hindleg muscle a strongly reduced phosphocreatine (PCr) content was shown. In addition, a signal of phosphorylated Gua at 0.5 ppm upfield of PCr was observed, with much higher intensity in muscle than in brain. This signal decreased when ischemia was applied to the muscle and recovered after ischemia was released. Overall, the in vivo (31)P and (1)H MR spectroscopy of GAMT-/- mice is similar to that of human GAMT deficiency. This opens up new avenues for the fundamental study of tissue-type dependence of creatine synthesis and transport and for diagnostic and therapeutic aspects of creatine deficiencies in humans.


Assuntos
Encéfalo/metabolismo , Creatina/deficiência , Glicina/análogos & derivados , Espectroscopia de Ressonância Magnética , Metiltransferases/deficiência , Músculo Esquelético/metabolismo , Animais , Química Encefálica , Glicina/metabolismo , Guanidinoacetato N-Metiltransferase , Membro Posterior , Humanos , Erros Inatos do Metabolismo/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout
20.
Magn Reson Med ; 50(3): 468-73, 2003 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-12939753

RESUMO

Magnetization transfer (MT) between the mobile (MR-visible) spin pool and immobile (MR-invisible) spin pool of creatine (Cr) was studied on a clinical 1.5 T MR scanner in human skeletal muscle using continuous wave (CW) pre-irradiation as the saturation method for the immobile pool. For this purpose, only slight modifications to the MR system were made. A specially designed electronic circuit was used to couple a CW amplifier to the RF channel of the scanner. The CW pulse power (gammaB(2)/2pi) and pulse length were determined to be approximately 550 Hz and 3 s, respectively, for optimal signal attenuation of the Cr methyl signal. The bound Cr fraction in human gastrocnemius muscle was determined to be 0.4-1.3% using a two-pool exchange model function to describe the MT effect.


Assuntos
Creatina/metabolismo , Espectroscopia de Ressonância Magnética/métodos , Músculo Esquelético/metabolismo , Desenho de Equipamento , Humanos , Perna (Membro) , Magnetismo
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