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1.
J Plast Reconstr Aesthet Surg ; 75(9): 3293-3303, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35725956

RESUMO

Conventional X-ray imaging is usually the first diagnostic assessment after clinical examination in case of suspected scapholunate interosseous ligament (SLIL) injury. Nevertheless, there is no consensus on the norms of the scapholunate distance (SLD) or carpal angles. Therefore, we aimed to determine the utility of static and dynamic radiographs in the diagnostic of an SLIL injury in comparison with the reference standard arthroscopy. We retrospectively analyzed the preoperative X-ray series and surgical records of arthroscopies of 414 patients. Radiological assessment included conventional static X-rays in the posteroanterior (PA) projection, clenched fist views in ulnar and radial deviations, in which the SLD at the midportion of the scapholunate (SL) joint was measured. The scapholunate angle (SLA) and radiolunate angle (RLA) were measured on lateral wrist radiographs. Receiver operating characteristic (ROC) curves were created to determine possible cut-off points of the radiological indices for the diagnosis of SLIL injury. Further, a logistic regression with the parameters having the highest area under the curve (AUC) was calculated. We found that SLD in ulnar inclination (AUC= 0.774), SLD in PA projection (AUC = 0.748), and SLA (AUC = 0.737) had the highest diagnostic value. The AUC of the combination of these three parameters was 0.822 for all patients with any SLIL lesion and 0.850 for patients with SLIL lesions of 3-4 Geissler grade. Further investigation of SLIL pathology would be appropriate in the case of SLD in ulnar inclination of 2.7 mm, SLD in PA projection of 1.9 mm and SLA of 63°.


Assuntos
Osso Semilunar , Osso Escafoide , Traumatismos do Punho , Artroscopia , Humanos , Ligamentos Articulares/diagnóstico por imagem , Osso Semilunar/diagnóstico por imagem , Estudos Retrospectivos , Osso Escafoide/diagnóstico por imagem , Traumatismos do Punho/diagnóstico por imagem , Traumatismos do Punho/cirurgia , Articulação do Punho/diagnóstico por imagem , Raios X
2.
Orthopade ; 49(10): 849-859, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32944784

RESUMO

The field of musculoskeletal diagnostics and personalized medicine has undergone a revolutionary transformation due to a deeper understanding of skeletal biomechanics and due to technological advancements. Analogous to this transformation, our understanding of spinopelvic conditions has experienced a paradigm shift in terms of both static and dynamic changes in spinopelvic pathologies and enabled a more accurate delineation of the drivers of disability. The purpose of this review is to describe the standard and state of the art of preoperative diagnostic and planning methods for common spinopelvic pathologies and to discuss both the added clinical value and limitations. The rationale is to accelerate the accurate and timely diagnosis and as well as the efficient and safe preoperative workflow.


Assuntos
Pelve/diagnóstico por imagem , Coluna Vertebral/diagnóstico por imagem , Humanos , Pelve/patologia , Período Pré-Operatório , Coluna Vertebral/patologia
3.
Scand J Med Sci Sports ; 26(10): 1200-8, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26369754

RESUMO

The purpose of this study was to evaluate temporal alterations of the Achilles tendon volume and hydration state after cross-country-running. Achilles tendons of six untrained participants were examined on a 3T MR-scanner before running, immediately afterwards, and in the following 24, 48, and 72 h. Using a 3D-UTE sequence, caudal (CA) and cranial (CR) mid-portion tendon areas were examined with off-resonance saturation ratios (OSR) and T2* relaxation times. Tendon volume was measured with a self-written Matlab-based automated contour detection algorithm (AVAT) in submillimeter T2-weighted MR images. A significant influence of running in caudal (P = 0.017) and cranial OSR values (P = 0.001), tendon volume (P = 0.024), and cranial T2* measurements (P = 0.046), but not in caudal T2* values (P = 0.298) were found. In detail, mean individual OSR and tendon volume measurements demonstrated a similar but inverted course in their values after exercise: initially, OSR values increased after running (and tendon volume decreased), while subsequently a decrease of OSR values (with an increase of tendon volume) could be observed. OSR and tendon volume measurements are able to detect a physiological response of tendons to a mechanical stimulus. After a transient decrease of free water in the Achilles tendon, an increase with a maximum free water content 48 h after ankle loading and a tendency toward normalization after 72 h was found.


Assuntos
Tendão do Calcâneo/anatomia & histologia , Tendão do Calcâneo/metabolismo , Articulação do Tornozelo/fisiologia , Imageamento por Ressonância Magnética , Corrida/fisiologia , Suporte de Carga/fisiologia , Tendão do Calcâneo/diagnóstico por imagem , Adulto , Seguimentos , Humanos , Imageamento por Ressonância Magnética/métodos , Masculino , Tamanho do Órgão , Fatores de Tempo , Água/metabolismo
4.
Rofo ; 187(11): 1003-10, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26090729

RESUMO

PURPOSE: The purpose of this study was to evaluate whether gravitational interstitial fluid accumulation in healthy subjects has an impact on off-resonance saturation ratios (OSR) or the volume of the Achilles tendon after a prolonged time of reduced levels of physical activity. MATERIALS AND METHODS: 7 healthy volunteers were repeatedly investigated on 3 consecutive days on a 3 T whole body MR scanner using an ultrashort echo time (UTE) imaging sequence with a Gaussian off-resonance saturation pulse at a frequency offset of 2000 Hz to calculate OSR values. For accurate volumetric quantification of the Achilles tendon, a newly developed contour detection snake algorithm was applied on high-resolution isotropic T2-weighted SPACE sequence datasets. Single-measure intraclass correlation coefficients (ICC) were calculated to estimate test-retest reliability. RESULTS: For OSR and tendon volume measurements on three consecutive days, excellent reproducibility could be achieved with ICC values above 0.96 and 0.97, respectively. Comparing the results of all three days, a statistically significant mean individual percentage decrease (- 4.1  ±â€Š1.5 %; p = 0.001) of calculated tendon OSR values was found for the evening measurements. No statistically significant difference between tendon volumes in the morning and the evening could be detected (p = 0.589). CONCLUSION: The results of this in-vivo study demonstrate a significant influence of gravitational interstitial fluid accumulation after reduced physical activity on OSR values in the Achilles tendon, but not on tendon volume. Taken together with the demonstrated excellent reproducibility, these findings are important for future studies investigating temporal changes of the Achilles tendon microstructure.


Assuntos
Tendão do Calcâneo/anatomia & histologia , Ritmo Circadiano/fisiologia , Líquido Extracelular/fisiologia , Deslocamentos de Líquidos Corporais/fisiologia , Gravitação , Interpretação de Imagem Assistida por Computador/métodos , Imageamento Tridimensional/métodos , Imageamento por Ressonância Magnética/métodos , Imagem Corporal Total/métodos , Adulto , Feminino , Humanos , Masculino , Atividade Motora/fisiologia , Distribuição Normal , Tamanho do Órgão/fisiologia , Valores de Referência
5.
Rofo ; 185(11): 1041-55, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23888478

RESUMO

In the past decades the incidence of acute and chronic disorders of the Achilles tendon associated with sport-induced overuse has steadily increased. Besides acute complete or partial ruptures, achillodynia (Achilles tendon pain syndrome), which is often associated with tendon degeneration, represents the most challenging entity regarding clinical diagnostics and therapy. Therefore, the use of imaging techniques to differentiate tendon disorders and even characterize structure alterations is of growing interest. This review article discusses the potential of different imaging techniques with respect to the diagnosis of acute and chronic tendon disorders. In this context, the most commonly used imaging techniques are magnetic resonance imaging (MRI), B-mode ultrasound, and color-coded Doppler ultrasound (US). These modalities allow the detection of acute tendon ruptures and advanced chronic tendon disorders. However, the main disadvantages are still the low capabilities in the detection of early-stage degeneration and difficulties in the assessment of treatment responses during follow-up examinations. Furthermore, differentiation between chronic partial ruptures and degeneration remains challenging. The automatic contour detection and texture analysis may allow a more objective and quantitative interpretation, which might be helpful in the monitoring of tendon diseases during follow-up examinations. Other techniques to quantify tendon-specific MR properties, e. g. based on ultrashort echo time (UTE) sequences, also seem to have great potential with respect to the precise detection of degenerative tendon disorders and their differentiation at a very early stage.


Assuntos
Tendão do Calcâneo/lesões , Tendão do Calcâneo/patologia , Transtornos Traumáticos Cumulativos/patologia , Aumento da Imagem/métodos , Imageamento por Ressonância Magnética/métodos , Traumatismos dos Tendões/patologia , Doença Aguda , Doença Crônica , Humanos , Ruptura/patologia
6.
Eur Radiol ; 23(2): 588-95, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22898936

RESUMO

OBJECTIVES: To evaluate stent lumen assessment of various commonly used and newly developed stents for the superficial femoral artery (SFA) using MR angiography (MRA) at 1.5 and 3 T. METHODS: Eleven nitinol stents and one cobalt-chromium stent were compared regarding stent lumen visualisation using a common three-dimensional MRA sequence. Maximum visible stent lumen width and contrast ratio were analysed in three representative slices for each stent type. A scoring system for lumen visualisation was applied. RESULTS: Nitinol stents showed significantly better performance than the cobalt chromium stent (P < 0.05) at 1.5 and 3 T. Maximum visible stent lumen ranged between 43.4 and 95.5 %, contrast ratio between 7.2 and 110.6 %. Regarding both field strengths, seven of the nitinol stents were classified as "suitable". Three nitinol stents were "limited", and one nitinol stent and the cobalt chromium stent were "not suitable". CONCLUSIONS: Intraluminal loss of signal and artefacts of most of the SFA stents do not markedly limit assessment of stent lumen by MRA at 1.5 and 3 T. MRA can thus be considered a valid technique for detection of relevant in-stent restenosis. Applied field strength does not strongly influence stent lumen assessment in general, but proper choice of field strength might be helpful.


Assuntos
Ligas , Cobalto , Imageamento Tridimensional , Angiografia por Ressonância Magnética/métodos , Stents , Meios de Contraste , Feminino , Artéria Femoral , Humanos , Técnicas In Vitro , Masculino , Teste de Materiais , Imagens de Fantasmas , Desenho de Prótese , Estatísticas não Paramétricas
7.
Eur J Radiol ; 81(12): 4173-8, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22795844

RESUMO

AIM: To describe computed tomography (CT)-imaging findings in human metapneumovirus (HMPV)-related pulmonary infection as well as their temporal course and to analyze resemblances/differences to pulmonary infection induced by the closely related respiratory-syncytial-virus (RSV) in immunocompromised patients. MATERIALS AND METHODS: Chest-CT-scans of 10 HMPV PCR-positive patients experiencing pulmonary symptoms were evaluated retrospectively with respect to imaging findings and their distribution and results were then compared with data acquired in 13 patients with RSV pulmonary infection. Subsequently, we analyzed the course of chest-findings in HMPV patients. RESULTS: In HMPV, 8/10 patients showed asymmetric pulmonary findings, whereas 13/13 patients with RSV-pneumonia presented more symmetrical bilateral pulmonary infiltrates. Image analysis yielded in HMPV patients following results: ground-glass-opacity (GGO) (n=6), parenchymal airspace consolidations (n=5), ill-defined nodular-like centrilobular opacities (n=9), bronchial wall thickening (n=8). In comparison, results in RSV patients were: GGO (n=10), parenchymal airspace consolidations (n=9), ill-defined nodular-like centrilobular opacities (n=10), bronchial wall thickening (n=4). In the course of the disease, signs of acute HMPV interstitial pneumonia regressed transforming temporarily in part into findings compatible with bronchitis/bronchiolitis. CONCLUSIONS: Early chest-CT findings in patients with HMPV-related pulmonary symptoms are compatible with asymmetric acute interstitial pneumonia accompanied by signs of bronchitis; the former transforming with time into bronchitis and bronchiolitis before they resolve. On the contrary, RSV-induced pulmonary infection exhibits mainly symmetric acute interstitial pneumonia.


Assuntos
Hospedeiro Imunocomprometido , Pneumopatias/diagnóstico por imagem , Pneumopatias/imunologia , Metapneumovirus , Infecções por Vírus Respiratório Sincicial/diagnóstico por imagem , Infecções por Vírus Respiratório Sincicial/imunologia , Tomografia Computadorizada por Raios X/métodos , Adulto , Idoso , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Reprodutibilidade dos Testes , Infecções por Vírus Respiratório Sincicial/virologia , Sensibilidade e Especificidade
8.
Eur J Radiol ; 81(7): 1612-7, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21543175

RESUMO

PURPOSE: Achilles tendinopathy has been reported to be frequently associated with increasing volume of the tendon. This work aims at reliable and accurate volumetric quantification of the Achilles tendon using a newly developed contour detection algorithm applied on high resolution MRI data sets recorded at 3T. MATERIALS AND METHODS: A total of 26 healthy tendons and 4 degenerated tendons were examined for this study. Automated identification (AI) of tendon boundaries was performed in transverse slices with isotropic resolution (0.8mm) gained with a T2-weighted SPACE sequence at 3T. For AI a snake algorithm was applied and compared to manual tracing (MT). RESULTS: AI was feasible in all examined tendons without further correction. AI of both tendons was performed in each participant within 2 min (2 × 37 slices) compared to MT lasting 20 min. MT and AI showed excellent agreement and correlation (R(2) = 0.99, p<0.0001). AI provided a reduction of measurement error (0.4 cm(3) vs. 0.5 cm(3)) and coefficient of variation (1% vs. 2%). DISCUSSION: Compared to MT the AI allows assessment of tendon volumes in highly resolved MRI data in a more accurate and reliable time-saving way. Therefore automated volume detection is seen as a helpful clinical tool for evaluation of small volumetric changes of the Achilles tendon.


Assuntos
Tendão do Calcâneo/patologia , Imageamento Tridimensional , Imageamento por Ressonância Magnética/métodos , Tendinopatia/diagnóstico , Tendão do Calcâneo/anatomia & histologia , Adulto , Algoritmos , Feminino , Humanos , Masculino , Reprodutibilidade dos Testes , Estatísticas não Paramétricas
9.
Rofo ; 183(11): 1043-50, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21986866

RESUMO

PURPOSE: Magnetization transfer contrast (MTC) imaging provides insight into interactions between free and bounded water. Newly developed ultrashort echo time (UTE) sequences implemented on whole-body magnetic resonance (MR) scanners allow MTC imaging in tissues with extremely fast signal decay such as tendons. The aim of this study was to develop a technique for the quantification of the MT effect in healthy Achilles tendons in-vivo at 3 Tesla. MATERIALS AND METHODS: 16 normal tendons of volunteers with no history of tendinopathy were examined using a 3D-UTE sequence with a rectangular on-resonant excitation pulse and a Fermi-shaped off-resonant MT preparation pulse. The frequency of the MT pulse was varied from 1 to 5 kHz. MT effects were calculated in terms of the MT ratio (MTR) between measurements without and with MT preparation. Direct saturation effects of MT preparation on the signal intensity were evaluated using numerical simulation of Bloch equations. One patient with tendinopathy was examined to exemplarily show changes of MTR under pathologic conditions. RESULTS: Calculation of MTR data was feasible in all examined tendons and showed a decrease from 0.53 ± 0.05 to 0.25 ± 0.03 (1 kHz to 5 kHz) for healthy volunteers. Evaluation of variation with gender and dominance of ankle revealed no significant differences (p > 0.05). In contrast, the patient with confirmed tendinopathy showed MTR values between 0.36 (1 kHz) and 0.19 (5 kHz). CONCLUSION: MT effects in human Achilles tendons can be reliably assessed in-vivo using a 3D UTE sequence at 3 T. All healthy tendons showed similar MTR values (coefficient of variation 10.0 ± 1.2 %). The examined patient showed a clearly different MT effect revealing a changed microstructure in the case of tendinopathy.


Assuntos
Tendão do Calcâneo/patologia , Processamento de Imagem Assistida por Computador/métodos , Imageamento Tridimensional/métodos , Imageamento por Ressonância Magnética/métodos , Tendinopatia/diagnóstico , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Análise Numérica Assistida por Computador , Valores de Referência , Sensibilidade e Especificidade , Tendinopatia/patologia , Adulto Jovem
10.
Nanotechnology ; 21(38): 385703, 2010 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-20798462

RESUMO

An approach for tailoring the magnetic properties by ion irradiation of granular perpendicular CoCrPt:SiO(2) films grown on silica particles with sizes down to 10 nm was investigated. The as-prepared samples reveal an intriguing scaling dependence of the coercive field and remnant magnetization: both parameters are found to decrease with decreasing particle size. However, Co(+) irradiation at a low fluence of 0.5 x 10(14) cm(-2) already results in an opposite scaling behavior. It is assumed that this modification is due to the enhancement of the intergranular magnetic exchange coupling of the granular CoCrPt:SiO(2) film initiated by Co(+) irradiation resulting in a modified reversal behavior. Further increase of the irradiation fluence beyond 1.6 x 10(14) ions cm(-2) leads to a degradation of the magnetic layer properties, lowering the remnant magnetization and the coercive field in the easy-axis direction. Moreover, the local magnetic properties of the samples were analyzed by magnetic force microscopy revealing magnetic multi-domain cap structures.

11.
J Chem Phys ; 124(18): 184702, 2006 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-16709126

RESUMO

Intense nonequilibrium femtosecond laser excitation of gold nanoparticles in water leads to a transient heating of the nanoparticles, which decays via heat transfer to the water phase. It is shown that the water temperature rises to near the critical temperature and the water undergoes an explosive evaporation in the subnanosecond range. The formation of vapor bubbles shows a threshold dependence on laser fluence. The nascent nanoscale vapor bubbles change the heat dissipation drastically. The nanoscale structure is resolved directly with a combination of x-ray scattering methods sensitive to the particle lattice expansion and the change in the water structure factor.

12.
J Forensic Sci ; 45(2): 471-3, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10782976

RESUMO

A suicide committed with a power saw is an uncommonly reported incident: and the use of a chain saw in particular has been extremely rare. We report two cases of suicide that were committed by using chain saws. In each case the victim had a history of depression or of a prior suicide attempt, and applied the chain saw to his neck to kill himself. We summarize the findings of the two death investigations and the world literature pertaining to suicide committed with power saws.


Assuntos
Lesões do Pescoço/etiologia , Suicídio , Acidentes de Trabalho , Adulto , Idoso , Transtorno Depressivo , Medicina Legal , Humanos , Masculino
13.
J Biotechnol ; 77(1): 65-80, 2000 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-10674215

RESUMO

Potato plants (Solanum tuberosum L., cv. Désirée) were transformed with the polyphosphate kinase gene from Escherichia coli fused to the leader sequence of the ferredoxin oxidoreductase gene (FNR) from Spinacea oleracea under the control of the leaf specific St-LS1 promoter to introduce a novel phosphate pool in the chloroplasts of green tissues. Transgenic plants (cpPPK) in tissue culture developed necrotic lesions in older leaves and showed earlier leaf senescence while greenhouse plants showed no noticeable phenotype. Leaves of cpPPK plants contained less starch but higher concentrations of soluble sugars. The presence of polyphosphate in cpPPK leaves was demonstrated by toluidine blue staining and unambiguously verified and quantified by in vitro 31P-NMR of extracts. Polyphosphate accumulated during leaf development from 0.06 in juvenile leaves to 0.83 mg P g-1 DW in old leaves and had an average chain length of 18 residues in mature leaves. In situ 31P-NMR on small leaf pieces perfused with well-oxygenated medium showed only 0.036 mg P g-1 DW polyphosphate that was, however, greatly increased upon treatment with 50 mM ammonium sulfate at pH 7.3. This phenomenon along with a yield of 0.47 mg P g-1 DW polyphosphate from an extract of the same leaf material suggests that 93% of the polyphosphate pool is immobile. This conclusion is substantiated by the observation that no differences in polyphosphate pool sizes could be discerned between darkened and illuminated leaves, leaves treated with methylviologen or anaerobis and control leaves, treatments causing a change in the pool of ATP available for polyPi synthesis. Results are discussed in the context of the chelating properties of polyphosphates for cations and its consequences for the partitioning of photoassimilate between starch and soluble sugars.


Assuntos
Metabolismo dos Carboidratos , Cloroplastos/genética , Fosfotransferases (Aceptor do Grupo Fosfato)/genética , Polifosfatos/metabolismo , Solanum tuberosum/genética , Animais , Compartimento Celular/fisiologia , Cloroplastos/química , Cloroplastos/enzimologia , Corantes , Etanol , Espectroscopia de Ressonância Magnética , Isótopos de Fósforo , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Polifosfatos/análise , Proteínas Recombinantes de Fusão/metabolismo , Solventes , Cloreto de Tolônio , Transgenes/fisiologia
14.
Planta ; 209(2): 230-238, 1999 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-10436226

RESUMO

In order to examine whether alterations in the supply of precursor molecules into the starch biosynthetic pathway affected various characteristics of the starch, starch was isolated from potato (Solanum tuberosum L.) tubers containing reduced amounts of the enzyme ADP-glucose pyrophosphorylase (AGPase). It was found that although the type of crystalline polymorph in the starch was not altered, the amylose content was severely reduced. In addition, amylopectin from the transgenic plants accumulated more relatively short chains than that from control plants and the sizes of starch granules were reduced. The starch granules from the transgenic plants contained a greater amount of granule-bound starch synthase enzyme, which led to an increase in the maximum activity of the enzyme per unit starch tested. The K(m) for ADP-glucose was, at most, only slightly altered in the transgenic lines. Potato plants containing reduced AGPase activity were also transformed with a bacterial gene coding for AGPase to test whether this enzyme can incorporate phosphate monoesters into amylopectin. A slight increase in phosphate contents in the starch in comparison with the untransformed control was found, but not in comparison with starch from the line with reduced AGPase activity into which the bacterial gene was transformed.Key words: ADP-glucose pyrophosphorylase. Amylopectin structure. Amylose. Solanum (starch. tuber). Starch granule size. Starch phosphorylation

15.
Planta ; 208(4): 503-11, 1999 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10420646

RESUMO

Three isoforms of starch synthase (SS) were shown to be present in soluble potato tuber extracts by activity staining after native gel electrophoresis. A cDNA encoding SSI from rice was used as a probe to clone a corresponding cDNA from potato. The deduced amino acid sequence identified the protein as an SS from potato with an M(r) of 70.6 kDa for the immature enzyme including its transit peptide. This novel isoform was designated SSI. An analysis of the expression pattern of the gene indicated that SSI is predominantly expressed in sink and source leaves, and, to a lower extent in tubers. In several independent transgenic potato lines, where the expression of SSI was repressed using the antisense approach, the activity of a specific SS isoform was reduced to non-detectable levels as determined through activity staining after native gel electrophoresis. The reduction in the amount of this isoform of SS did not lead to any detectable changes in starch structure, probably due to the fact that this isoform only represents a minor activity in potato tubers.


Assuntos
Solanum tuberosum/genética , Sintase do Amido/genética , Sequência de Aminoácidos , Clonagem Molecular , DNA Complementar , Dados de Sequência Molecular , Folhas de Planta/enzimologia , Homologia de Sequência de Aminoácidos , Solanum tuberosum/enzimologia
16.
Plant J ; 10(6): 981-91, 1996 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9011082

RESUMO

Three isoforms of starch synthase were shown to be present in soluble potato tuber extracts by activity staining after native gel electrophoresis. An antibody directed against a domain conserved in starch synthases was used to clone a cDNA for one of these isoforms by screening a tuber-specific expression library. A partial cDNA of 2.6 kbp was obtained and used to isolate a full-length cDNA of 4167 bp. The deduced amino acid sequence identifies the protein as a novel type of starch synthase from potato with a molecular mass of 139.2 kDa for the immature enzyme including its transit peptide. This novel isoform was designated SS III. An analysis of the expression pattern of the gene indicates that SS III is equally expressed in tubers of different developmental stages as well as in sink and source leaves. In several independent transgenic potato lines, where the expression of SS III was repressed using the antisense approach, the activity of a specific starch synthase isoform was reduced to non-detectable levels as determined through activity staining after native gel electrophoresis. The reduction of this isoform of starch synthase leads to the synthesis of a structurally modified starch in the transgenic plants: there is a drastic change in granule morphology and an increased level of covalently linked phosphate.


Assuntos
Isoenzimas/genética , Proteínas de Plantas/genética , Solanum tuberosum/genética , Sintase do Amido/genética , Sequência de Aminoácidos , Elementos Antissenso (Genética) , Evolução Biológica , Clonagem Molecular , DNA Complementar/genética , Expressão Gênica , Regulação da Expressão Gênica de Plantas , Isoenzimas/classificação , Dados de Sequência Molecular , Proteínas de Plantas/classificação , Plantas Geneticamente Modificadas , Análise de Sequência de DNA , Solanum tuberosum/enzimologia , Amido/química , Sintase do Amido/classificação , Distribuição Tecidual
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