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1.
Cancer Gene Ther ; 30(3): 497-506, 2023 03.
Artículo en Inglés | MEDLINE | ID: mdl-36494580

RESUMEN

Downregulation of cell-cell adhesion and increased motility are prerequisites for the metastasis of cancer cells. We have recently shown that downregulation of the tight junction adapter protein Pals1 in colorectal cancer cells results in an increase of cell migration, invasion, and metastasis due to the enhanced activation of Arf6 and Rac1. We now reveal a redundancy between the Arf6-GAP SMAP1 and Pals1 in regulating Arf6 activity and thereby Rac1-dependent cell migration. The gene encoding SMAP1 is frequently disrupted in microsatellite instable colorectal cancer specimen and cell lines. In cells expressing SMAP1, deletion of Pals1 leads to disturbed formation of tight junctions but has no impact on Arf6 activity and cell migration. In contrast, inactivation of both SMAP1 and Pals1 results in enhanced Arf6/Rac1 activity and increased cell migration and invasion. Furthermore, analyzing patient cohorts, we found a significant decrease in patient's survival when both genes were downregulated, in contrast to cases, when expression of only one of both genes was affected. Taken together, we identified a redundancy between SMAP1 and Pals1 in the regulation of activation of Arf6/Rac1, thereby controlling cell migration, invasion, and metastasis of colorectal cancer cells.


Asunto(s)
Factores de Ribosilacion-ADP , Neoplasias Colorrectales , Humanos , Factores de Ribosilacion-ADP/genética , Factores de Ribosilacion-ADP/metabolismo , Línea Celular , Movimiento Celular/genética , Neoplasias Colorrectales/genética , Proteínas Activadoras de GTPasa/genética , Proteínas Activadoras de GTPasa/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Proteína de Unión al GTP rac1/genética , Proteína de Unión al GTP rac1/metabolismo
2.
Z Gastroenterol ; 60(11): 1668-1677, 2022 Nov.
Artículo en Alemán | MEDLINE | ID: mdl-35297030

RESUMEN

In the work-up of chronic enteropathies an underlying inborn error of immunity (IEI) should be considered in certain cases. IEI are rare, but approximately 10% of patients may present with symptoms of inflammatory bowel disease (IBD), which is a much more common entity. Patients with IEI associated IBD may show extraintestinal symptoms or signs, and are often refractory to conventional anti-inflammatory treatment. In case of early-onset bowel inflammation and other intestinal or extraintestinal manifestations, an IEI should be excluded. A small fraction of monogenic IEI can be amenable to targeted therapies, or even corrected by allogeneic stem cell transplantation. Therefore, early diagnosis is crucial. This paper shows examples of clinical - gastrointestinal as well as extraintestinal - signs and findings which require immunological and possibly genetic workup.


Asunto(s)
Enfermedades Inflamatorias del Intestino , Humanos , Enfermedades Inflamatorias del Intestino/diagnóstico , Enfermedades Inflamatorias del Intestino/terapia , Inflamación
3.
PeerJ ; 10: e12635, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35174011

RESUMEN

Experimental approaches are often used to better understand the mechanisms behind and consequences of post-mortem alteration on proxies for diet reconstruction. Dental microwear texture analysis (DMTA) is such a dietary proxy, using dental wear features in extant and extinct taxa to reconstruct feeding behaviour and mechanical food properties. In fossil specimens especially, DMTA can be biased by post-mortem alteration caused by mechanical or chemical alteration of the enamel surface. Here we performed three different dental surface alteration experiments to assess the effect of common taphonomic processes by simplifying them: (1) tumbling in sediment suspension to simulate fluvial transport, (2) sandblasting to simulate mechanical erosion due to aeolian sediment transport, (3) acid etching to simulate chemical dissolution by stomach acid. For tumbling (1) we found alteration to be mainly dependent on sediment grain size fraction and that on specimens tumbled with sand fractions mainly post-mortem scratches formed on the dental surface, while specimens tumbled with a fine-gravel fraction showed post-mortem formed dales. Sandblasting (2) with loess caused only negligible alteration, however blasting with fine sand quartz particles resulted in significant destruction of enamel surfaces and formation of large post-mortem dales. Acid etching (3) using diluted hydrochloric acid solutions in concentrations similar to that of predator stomachs led to a complete etching of the whole dental surface, which did not resemble those of teeth recovered from owl pellets. The experiments resulted in post-mortem alteration comparable, but not identical to naturally occurring post-mortem alteration features. Nevertheless, this study serves as a first assessment and step towards further, more refined taphonomic experiments evaluating post-mortem alteration of dental microwear texture (DMT).


Asunto(s)
Desgaste de los Dientes , Diente , Humanos , Arena , Alimentos , Esmalte Dental
4.
Mol Cancer ; 20(1): 74, 2021 05 04.
Artículo en Inglés | MEDLINE | ID: mdl-33941200

RESUMEN

Loss of apical-basal polarity and downregulation of cell-cell contacts is a critical step during the pathogenesis of cancer. Both processes are regulated by the scaffolding protein Pals1, however, it is unclear whether the expression of Pals1 is affected in cancer cells and whether Pals1 is implicated in the pathogenesis of the disease.Using mRNA expression data and immunostainings of cancer specimen, we show that Pals1 is frequently downregulated in colorectal cancer, correlating with poorer survival of patients. We further found that Pals1 prevents cancer cell metastasis by controlling Rac1-dependent cell migration through inhibition of Arf6, which is independent of the canonical binding partners of Pals1. Loss of Pals1 in colorectal cancer cells results in increased Arf6 and Rac1 activity, enhanced cell migration and invasion in vitro and increased metastasis of transplanted tumor cells in mice. Thus, our data reveal a new function of Pals1 as a key inhibitor of cell migration and metastasis of colorectal cancer cells. Notably, this new function is independent of the known role of Pals1 in tight junction formation and apical-basal polarity.


Asunto(s)
Factor 6 de Ribosilación del ADP/metabolismo , Neoplasias Colorrectales/patología , Proteínas de la Membrana/metabolismo , Nucleósido-Fosfato Quinasa/metabolismo , Proteína de Unión al GTP rac1/metabolismo , Animales , Movimiento Celular/fisiología , Células HCT116 , Xenoinjertos , Humanos , Ratones , Invasividad Neoplásica/patología
5.
Proc Natl Acad Sci U S A ; 117(36): 22264-22273, 2020 09 08.
Artículo en Inglés | MEDLINE | ID: mdl-32839331

RESUMEN

Food processing wears down teeth, thus affecting tooth functionality and evolutionary success. Other than intrinsic silica phytoliths, extrinsic mineral dust/grit adhering to plants causes tooth wear in mammalian herbivores. Dental microwear texture analysis (DMTA) is widely applied to infer diet from microscopic dental wear traces. The relationship between external abrasives and dental microwear texture (DMT) formation remains elusive. Feeding experiments with sheep have shown negligible effects of dust-laden grass and browse, suggesting that intrinsic properties of plants are more important. Here, we explore the effect of clay- to sand-sized mineral abrasives (quartz, volcanic ash, loess, kaolin) on DMT in a controlled feeding experiment with guinea pigs. By adding 1, 4, 5, or 8% mineral abrasives to a pelleted base diet, we test for the effect of particle size, shape, and amount on DMT. Wear by fine-grained quartz (>5/<50 µm), loess, and kaolin is not significantly different from the abrasive-free control diet. Fine silt-sized quartz (∼5 µm) results in higher surface anisotropy and lower roughness (polishing effect). Coarse-grained volcanic ash leads to significantly higher complexity, while fine sands (130 to 166 µm) result in significantly higher roughness. Complexity and roughness values exceed those from feeding experiments with guinea pigs who received plants with different phytolith content. Our results highlight that large (>95-µm) external silicate abrasives lead to distinct microscopic wear with higher roughness and complexity than caused by mineral abrasive-free herbivorous diets. Hence, high loads of mineral dust and grit in natural diets might be identified by DMTA, also in the fossil record.


Asunto(s)
Alimentación Animal , Cobayas , Plantas , Abrasión de los Dientes/veterinaria , Desgaste de los Dientes/veterinaria , Animales , Dieta/veterinaria , Herbivoria , Tamaño de la Partícula , Abrasión de los Dientes/etiología
6.
Beilstein J Nanotechnol ; 5: 1944-65, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25383306

RESUMEN

PVP-capped silver nanoparticles with a diameter of the metallic core of 70 nm, a hydrodynamic diameter of 120 nm and a zeta potential of -20 mV were prepared and investigated with regard to their biological activity. This review summarizes the physicochemical properties (dissolution, protein adsorption, dispersability) of these nanoparticles and the cellular consequences of the exposure of a broad range of biological test systems to this defined type of silver nanoparticles. Silver nanoparticles dissolve in water in the presence of oxygen. In addition, in biological media (i.e., in the presence of proteins) the surface of silver nanoparticles is rapidly coated by a protein corona that influences their physicochemical and biological properties including cellular uptake. Silver nanoparticles are taken up by cell-type specific endocytosis pathways as demonstrated for hMSC, primary T-cells, primary monocytes, and astrocytes. A visualization of particles inside cells is possible by X-ray microscopy, fluorescence microscopy, and combined FIB/SEM analysis. By staining organelles, their localization inside the cell can be additionally determined. While primary brain astrocytes are shown to be fairly tolerant toward silver nanoparticles, silver nanoparticles induce the formation of DNA double-strand-breaks (DSB) and lead to chromosomal aberrations and sister-chromatid exchanges in Chinese hamster fibroblast cell lines (CHO9, K1, V79B). An exposure of rats to silver nanoparticles in vivo induced a moderate pulmonary toxicity, however, only at rather high concentrations. The same was found in precision-cut lung slices of rats in which silver nanoparticles remained mainly at the tissue surface. In a human 3D triple-cell culture model consisting of three cell types (alveolar epithelial cells, macrophages, and dendritic cells), adverse effects were also only found at high silver concentrations. The silver ions that are released from silver nanoparticles may be harmful to skin with disrupted barrier (e.g., wounds) and induce oxidative stress in skin cells (HaCaT). In conclusion, the data obtained on the effects of this well-defined type of silver nanoparticles on various biological systems clearly demonstrate that cell-type specific properties as well as experimental conditions determine the biocompatibility of and the cellular responses to an exposure with silver nanoparticles.

7.
PLoS One ; 9(5): e97477, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24828251

RESUMEN

The establishment of kleptoplasty (retention of "stolen plastids") in the digestive tissue of the sacoglossan Elysia chlorotica Gould was investigated using transmission electron microscopy. Cellular processes occurring during the initial exposure to plastids were observed in laboratory raised animals ranging from 1-14 days post metamorphosis (dpm). These observations revealed an abundance of lipid droplets (LDs) correlating to plastid abundance. Starvation of animals resulted in LD and plastid decay in animals <5 dpm that had not yet achieved permanent kleptoplasty. Animals allowed to feed on algal prey (Vaucheria litorea C. Agardh) for 7 d or greater retained stable plastids resistant to cellular breakdown. Lipid analysis of algal and animal samples supports that these accumulating LDs may be of plastid origin, as the often algal-derived 20∶5 eicosapentaenoic acid was found in high abundance in the animal tissue. Subsequent culturing of animals in dark conditions revealed a reduced ability to establish permanent kleptoplasty in the absence of photosynthetic processes, coupled with increased mortality. Together, these data support an important role of photosynthetic lipid production in establishing and stabilizing this unique animal kleptoplasty.


Asunto(s)
Gastrópodos/genética , Lípidos/genética , Plastidios/genética , Simbiosis/genética , Animales , Gastrópodos/metabolismo , Gotas Lipídicas/metabolismo , Fotosíntesis/genética , Plantas/genética , Plantas/metabolismo , Plastidios/metabolismo
8.
Mol Plant ; 7(1): 45-57, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-24043709

RESUMEN

Ferredoxin-NADP+ oxidoreductase (FNR), functioning in the last step of the photosynthetic electron transfer chain, exists both as a soluble protein in the chloroplast stroma and tightly attached to chloroplast membranes. Surface plasmon resonance assays showed that the two FNR isoforms, LFNR1 and LFNR2, are bound to the thylakoid membrane via the C-terminal domains of Tic62 and TROL proteins in a pH-dependent manner. The tic62 trol double mutants contained a reduced level of FNR, exclusively found in the soluble stroma. Although the mutant plants showed no visual phenotype or defects in the function of photosystems under any conditions studied, a low ratio of NADPH/NADP+ was detected. Since the CO2 fixation capacity did not differ between the tic62 trol plants and wild-type, it seems that the plants are able to funnel reducing power to most crucial reactions to ensure survival and fitness of the plants. However, the activity of malate dehydrogenase was down-regulated in the mutant plants. Apparently, the plastid metabolism is able to cope with substantial changes in directing the electrons from the light reactions to stromal metabolism and thus only few differences are visible in steady-state metabolite pool sizes of the tic62 trol plants.


Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/metabolismo , Ferredoxina-NADP Reductasa/deficiencia , Proteínas de la Membrana/genética , Proteínas de Transporte de Membrana/genética , Mutación , Fenotipo , Tilacoides/metabolismo , Secuencia de Aminoácidos , Arabidopsis/citología , Arabidopsis/genética , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/metabolismo , Cloroplastos/metabolismo , Proteínas de la Membrana/química , Proteínas de la Membrana/metabolismo , Proteínas de Transporte de Membrana/química , Proteínas de Transporte de Membrana/metabolismo , Datos de Secuencia Molecular , NADP/metabolismo , Oxidación-Reducción , Almidón/biosíntesis
9.
PLoS One ; 8(2): e57078, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23437316

RESUMEN

Mutations in the gene NPHS2 are the most common cause of hereditary steroid-resistant nephrotic syndrome. Its gene product, the stomatin family member protein podocin represents a core component of the slit diaphragm, a unique structure that bridges the space between adjacent podocyte foot processes in the kidney glomerulus. Dislocation and misexpression of slit diaphragm components have been described in the pathogenesis of acquired and hereditary nephrotic syndrome. However, little is known about mechanisms regulating cellular trafficking and turnover of podocin. Here, we discover a three amino acids-comprising motif regulating intracellular localization of podocin in cell culture systems. Mutations of this motif led to markedly reduced degradation of podocin. These findings give novel insight into the molecular biology of the slit diaphragm protein podocin, enabling future research to establish the biological relevance of podocin turnover and localization.


Asunto(s)
Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas de la Membrana/metabolismo , Dominios y Motivos de Interacción de Proteínas , Animales , Técnicas de Cultivo de Célula , Línea Celular , Membrana Celular/metabolismo , Endosomas/metabolismo , Humanos , Péptidos y Proteínas de Señalización Intracelular/química , Péptidos y Proteínas de Señalización Intracelular/genética , Microdominios de Membrana , Proteínas de la Membrana/química , Proteínas de la Membrana/genética , Ratones , Mutación , Transporte de Proteínas , Proteolisis
10.
Mol Plant ; 6(3): 817-29, 2013 May.
Artículo en Inglés | MEDLINE | ID: mdl-23204504

RESUMEN

Preprotein import into chloroplasts depends on macromolecular machineries in the outer and inner chloroplast envelope membrane (TOC and TIC). It was suggested that both machineries are interconnected by components of the intermembrane space (IMS). That is, amongst others, Tic22, of which two closely related isoforms exist in Arabidopsis thaliana, namely atTic22-III and atTic22-IV. We investigated the function of Tic22 in vivo by analyzing T-DNA insertion lines of the corresponding genes. While the T-DNA insertion in the individual genes caused only slight defects, a double mutant of both isoforms showed retarded growth, a pale phenotype under high-light conditions, a reduced import rate, and a reduction in the photosynthetic performance of the plants. The latter is supported by changes in the metabolite content of mutant plants when compared to wild-type. Thus, our results support the notion that Tic22 is directly involved in chloroplast preprotein import and might point to a particular importance of Tic22 in chloroplast biogenesis at times of high import rates.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Cloroplastos/metabolismo , Membranas Intracelulares/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis/genética , Clorofila/metabolismo , Cloroplastos/efectos de la radiación , Cloroplastos/ultraestructura , ADN Bacteriano/genética , Eliminación de Gen , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Técnicas de Inactivación de Genes , Genes de Plantas/genética , Genotipo , Membranas Intracelulares/efectos de la radiación , Membranas Intracelulares/ultraestructura , Luz , Proteínas de Transporte de Membrana/genética , Metaboloma/efectos de la radiación , Mutagénesis Insercional/genética , Fenotipo , Fotosíntesis/efectos de la radiación , Desarrollo de la Planta/genética , Desarrollo de la Planta/efectos de la radiación , Transporte de Proteínas/efectos de la radiación
11.
Integr Biol (Camb) ; 4(5): 480-93, 2012 May.
Artículo en Inglés | MEDLINE | ID: mdl-22402787

RESUMEN

The contribution of metabolism to heat stress may play a significant role in defining robustness and recovery of systems; either by providing the energy and metabolites required for cellular homeostasis, or through the generation of protective osmolytes. However, the mechanisms by which heat stress attenuation could be adapted through metabolic processes as a stabilizing strategy against thermal stress are still largely unclear. We address this issue through metabolomic and transcriptomic profiles for populations along a thermal cline where two seagrass species, Zostera marina and Zostera noltii, were found in close proximity. Significant changes captured by these profile comparisons could be detected, with a larger response magnitude observed in northern populations to heat stress. Sucrose, fructose, and myo-inositol were identified to be the most responsive of the 29 analyzed organic metabolites. Many key enzymes in the Calvin cycle, glycolysis and pentose phosphate pathways also showed significant differential expression. The reported comparison suggests that adaptive mechanisms are involved through metabolic pathways to dampen the impacts of heat stress, and interactions between the metabolome and proteome should be further investigated in systems biology to understand robust design features against abiotic stress.


Asunto(s)
Respuesta al Choque Térmico/fisiología , Metaboloma/fisiología , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Zosteraceae/fisiología , Adaptación Fisiológica/fisiología , Especificidad de la Especie , Zosteraceae/clasificación
12.
Plant Cell ; 23(6): 2087-105, 2011 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-21705644

RESUMEN

Selective pressure exerted by a massive decline in atmospheric CO(2) levels 55 to 40 million years ago promoted the evolution of a novel, highly efficient mode of photosynthetic carbon assimilation known as C(4) photosynthesis. C(4) species have concurrently evolved multiple times in a broad range of plant families, and this multiple and parallel evolution of the complex C(4) trait indicates a common underlying evolutionary mechanism that might be elucidated by comparative analyses of related C(3) and C(4) species. Here, we use mRNA-Seq analysis of five species within the genus Flaveria, ranging from C(3) to C(3)-C(4) intermediate to C(4) species, to quantify the differences in the transcriptomes of closely related plant species with varying degrees of C(4)-associated characteristics. Single gene analysis defines the C(4) cycle enzymes and transporters more precisely and provides new candidates for yet unknown functions as well as identifies C(4) associated pathways. Molecular evidence for a photorespiratory CO(2) pump prior to the establishment of the C(4) cycle-based CO(2) pump is provided. Cluster analysis defines the upper limit of C(4)-related gene expression changes in mature leaves of Flaveria as 3582 alterations.


Asunto(s)
Evolución Biológica , Carbono/química , Carbono/metabolismo , Flaveria/genética , Flaveria/metabolismo , Fotosíntesis/genética , Aminoácidos/metabolismo , Ciclo del Carbono/fisiología , Dióxido de Carbono/metabolismo , Isótopos de Carbono/química , Isótopos de Carbono/metabolismo , Cloroplastos/metabolismo , Análisis por Conglomerados , Transporte de Electrón , Flaveria/anatomía & histología , Flaveria/clasificación , Expresión Génica , Regulación de la Expresión Génica de las Plantas , Nitrógeno/metabolismo , Fotosíntesis/fisiología , Filogenia , Hojas de la Planta/química , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ribulosa-Bifosfato Carboxilasa/genética , Ribulosa-Bifosfato Carboxilasa/metabolismo , Transcriptoma
13.
J Biol Chem ; 286(13): 11382-90, 2011 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-21296880

RESUMEN

D-2-Hydroxyglutarate dehydrogenase (D-2HGDH) catalyzes the specific and efficient oxidation of D-2-hydroxyglutarate (D-2HG) to 2-oxoglutarate using FAD as a cofactor. In this work, we demonstrate that D-2HGDH localizes to plant mitochondria and that its expression increases gradually during developmental and dark-induced senescence in Arabidopsis thaliana, indicating an enhanced demand of respiration of alternative substrates through this enzymatic system under these conditions. Using loss-of-function mutants in D-2HGDH (d2hgdh1) and stable isotope dilution LC-MS/MS, we found that the D-isomer of 2HG accumulated in leaves of d2hgdh1 during both forms of carbon starvation. In addition to this, d2hgdh1 presented enhanced levels of most TCA cycle intermediates and free amino acids. In contrast to the deleterious effects caused by a deficiency in D-2HGDH in humans, d2hgdh1 and overexpressing lines of D-2HGDH showed normal developmental and senescence phenotypes, indicating a mild role of D-2HGDH in the tested conditions. Moreover, metabolic fingerprinting of leaves of plants grown in media supplemented with putative precursors indicated that D-2HG most probably originates during the catabolism of lysine. Finally, the L-isomer of 2HG was also detected in leaf extracts, indicating that both chiral forms of 2HG participate in plant metabolism.


Asunto(s)
Oxidorreductasas de Alcohol/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Lisina/metabolismo , Oxidorreductasas de Alcohol/genética , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Ciclo del Ácido Cítrico/fisiología , Glutaratos/metabolismo , Lisina/genética , Consumo de Oxígeno/fisiología
14.
Plant Physiol ; 155(1): 142-56, 2011 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-20543093

RESUMEN

C(4) photosynthesis involves alterations to the biochemistry, cell biology, and development of leaves. Together, these modifications increase the efficiency of photosynthesis, and despite the apparent complexity of the pathway, it has evolved at least 45 times independently within the angiosperms. To provide insight into the extent to which gene expression is altered between C(3) and C(4) leaves, and to identify candidates associated with the C(4) pathway, we used massively parallel mRNA sequencing of closely related C(3) (Cleome spinosa) and C(4) (Cleome gynandra) species. Gene annotation was facilitated by the phylogenetic proximity of Cleome and Arabidopsis (Arabidopsis thaliana). Up to 603 transcripts differ in abundance between these C(3) and C(4) leaves. These include 17 transcription factors, putative transport proteins, as well as genes that in Arabidopsis are implicated in chloroplast movement and expansion, plasmodesmatal connectivity, and cell wall modification. These are all characteristics known to alter in a C(4) leaf but that previously had remained undefined at the molecular level. We also document large shifts in overall transcription profiles for selected functional classes. Our approach defines the extent to which transcript abundance in these C(3) and C(4) leaves differs, provides a blueprint for the NAD-malic enzyme C(4) pathway operating in a dicotyledon, and furthermore identifies potential regulators. We anticipate that comparative transcriptomics of closely related species will provide deep insight into the evolution of other complex traits.


Asunto(s)
Cleome/genética , Cleome/fisiología , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Fotosíntesis/genética , Carbono/metabolismo , Genes de Plantas/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Modelos Biológicos , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Reacción en Cadena de la Polimerasa , ARN Mensajero/genética , Ribulosa-Bifosfato Carboxilasa/genética , Ribulosa-Bifosfato Carboxilasa/metabolismo , Especificidad de la Especie , Factores de Transcripción/genética , Factores de Transcripción/metabolismo
15.
BMC Cancer ; 10: 412, 2010 Aug 09.
Artículo en Inglés | MEDLINE | ID: mdl-20691121

RESUMEN

BACKGROUND: Breast cancer treatment is based on a combination of adjuvant chemotherapy followed by radiotherapy effecting intracellular signal transduction. With the tyrosine kinase inhibitors new targeted drugs are available. Imatinib mesylate is a selective inhibitor of bcr-abl, PRGFR alpha, beta and c-kit. The purpose of this study was to determine whether Imatinib has an influence on the effectiveness of radiotherapy in breast cancer cell lines and if a combination of imatinib with standard chemotherapy could lead to increased cytoreduction. METHODS: Colony-forming tests of MCF 7 and MDA MB 231 were used to study differences in cell proliferation under incubation with imatinib and radiation. Changes in expression and phosphorylation of target receptors were detected using western blot. Cell proliferation, migration and apoptosis assays were performed combining imatinib with doxorubicin. RESULTS: The combination of imatinib and radiotherapy showed a significantly stronger inhibition of cell proliferation compared to single radiotherapy. Differences in PDGFR expression could not be detected, but receptor phosphorylation was significantly inhibited when treated with imatinib. Combination of imatinib with standard chemotherapy lead to an additive effect on cell growth inhibition compared to single treatment. CONCLUSIONS: Imatinib treatment combined with radiotherapy leads in breast cancer cell lines to a significant benefit which might be influenced through inhibition of PDGFR phosphorylation. Combining imatinib with chemotherapy enhances cytoreductive effects. Further in vivo studies are needed to evaluate the benefit of Imatinib in combination with radiotherapy and chemotherapy on the treatment of breast cancer.


Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/radioterapia , Resistencia a Antineoplásicos/efectos de la radiación , Tolerancia a Radiación/efectos de los fármacos , Benzamidas , Western Blotting , Neoplasias de la Mama/patología , Adhesión Celular/efectos de los fármacos , Adhesión Celular/efectos de la radiación , Movimiento Celular/efectos de los fármacos , Movimiento Celular/efectos de la radiación , Proliferación Celular/efectos de los fármacos , Proliferación Celular/efectos de la radiación , Radioisótopos de Cobalto , Terapia Combinada , Doxorrubicina/administración & dosificación , Resistencia a Antineoplásicos/efectos de los fármacos , Femenino , Humanos , Mesilato de Imatinib , Radioisótopos de Iridio , Fosforilación/efectos de los fármacos , Fosforilación/efectos de la radiación , Piperazinas/administración & dosificación , Pirimidinas/administración & dosificación , Tolerancia a Radiación/efectos de la radiación , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/efectos de la radiación , Células Tumorales Cultivadas , Ensayo de Tumor de Célula Madre
16.
Biochem Mol Biol Educ ; 37(3): 170-7, 2009 May.
Artículo en Inglés | MEDLINE | ID: mdl-21567727

RESUMEN

The introduction of dwarfing traits into crops was a major factor in increased grain yields during the "Green Revolution." In most cases those traits were the consequence of altered synthesis or response to the gibberellin (GA) plant hormones. Our current understanding of GA synthesis and physiology has been facilitated by the characterization of mutants. To introduce concepts about GA hormone physiology and plant transformation in an undergraduate laboratory course we have used ga5, a semi-dwarf Arabidopsis mutant with reduced activity of GA 20-oxidase. In this laboratory exercise, Arabidopsis ga5 mutant plants are transformed by the floral-dip method using Agrobacterium tumefaciens carrying plasmid constructs conferring kanamycin resistance and containing the GA5 gene. Within 4 weeks, seeds of transformed plants can be easily screened by antibiotic resistance on plates. After transfer to soil the dwarf mutant plants transformed with a wild-type version of the gene show normal size. In addition to offering a visual understanding of the effect of GA on stem elongation, students learn additional techniques in this experiment, including PCR and agarose gel electrophoresis. This experiment is cost effective and can be completed within a 4-month term.

17.
Pediatr Crit Care Med ; 9(6): 629-35, 2008 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-18838934

RESUMEN

OBJECTIVE: Morbidity and mortality in preterm infants is significantly determined by the development of pulmonary complications. We thus investigated the impact of obstructive ventilatory disorders on lung function in very preterm infants with a history of respiratory distress syndrome and/or bronchopulmonary dysplasia using repeated body plethysmographic measurements before and after bronchodilation. DESIGN: Lung function, including effective airway resistance (Raw), specific conductance (SGaw), functional residual capacity (FRCbox), and total respiratory system compliance (Crs, multiple occlusion technique) was assessed in 27 preterm infants pound31 wks gestational age at a median postmenstrual age of 38 wks after mild oral sedation before and after inhalation of nebulized salbutamol (1.25 mg/2.5 mL; PARI JuniorBOY N) using the MasterScreen Baby Body (Jaeger, Hoechberg, Germany). RESULTS: In preterm infants median Raw was initially found to be within the normal range as determined for healthy term newborns, but decreased significantly after administration of salbutamol; SGaw changed accordingly. FRCbox was significantly reduced compared with healthy term newborns (16.6 vs. 19.6 mL/kg, mean) and decreased further after bronchodilation, whereas Crs was not significantly altered. CONCLUSIONS: This is the first report quantifying the important impact of obstructive ventilatory disorders on lung function in very preterm infants at term. Besides its important role in preterm lung function consecutive overinflation could furthermore be shown to mask reduction of lung volume in these infants. Thus, body plethysmographic measurements seem to be an important diagnostic tool in preterm infants at term before hospital discharge in order to quantify ventilation disorders and to define therapeutic strategies.


Asunto(s)
Obstrucción de las Vías Aéreas/fisiopatología , Pulmón/fisiopatología , Femenino , Humanos , Recién Nacido , Recien Nacido Prematuro , Masculino , Pletismografía , Pruebas de Función Respiratoria
18.
Plant Physiol ; 144(1): 32-42, 2007 May.
Artículo en Inglés | MEDLINE | ID: mdl-17351049

RESUMEN

Massively parallel sequencing of DNA by pyrosequencing technology offers much higher throughput and lower cost than conventional Sanger sequencing. Although extensively used already for sequencing of genomes, relatively few applications of massively parallel pyrosequencing to transcriptome analysis have been reported. To test the ability of this technology to provide unbiased representation of transcripts, we analyzed mRNA from Arabidopsis (Arabidopsis thaliana) seedlings. Two sequencing runs yielded 541,852 expressed sequence tags (ESTs) after quality control. Mapping of the ESTs to the Arabidopsis genome and to The Arabidopsis Information Resource 7.0 cDNA models indicated: (1) massively parallel pyrosequencing detected transcription of 17,449 gene loci providing very deep coverage of the transcriptome. Performing a second sequencing run only increased the number of genes identified by 10%, but increased the overall sequence coverage by 50%. (2) Mapping of the ESTs to their predicted full-length transcripts indicated that all regions of the transcript were well represented regardless of transcript length or expression level. Furthermore, short, medium, and long transcripts were equally represented. (3) Over 16,000 of the ESTs that mapped to the genome were not represented in the existing dbEST database. In some cases, the ESTs provide the first experimental evidence for transcripts derived from predicted genes, and, for at least 60 locations in the genome, pyrosequencing identified likely protein-coding sequences that are not now annotated as genes. Together, the results indicate massively parallel pyrosequencing provides novel information helpful to improve the annotation of the Arabidopsis genome. Furthermore, the unbiased representation of transcripts will be particularly useful for gene discovery and gene expression analysis of nonmodel plants with less complete genomic information. EST sequence accession numbers in GenBank are EH 795234 through EH 995233 and EL 000001 through EL 341852.


Asunto(s)
Arabidopsis/genética , ARN Mensajero/química , Análisis de Secuencia de ARN/métodos , Arabidopsis/metabolismo , Cloroplastos/genética , Mapeo Cromosómico , Etiquetas de Secuencia Expresada , Genes Mitocondriales , Genoma de Planta , Datos de Secuencia Molecular
19.
J Acoust Soc Am ; 116(3): 1781-92, 2004 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-15478445

RESUMEN

Numerous attempts have been made to find low-dimensional, formant-related representations of speech signals that are suitable for automatic speech recognition. However, it is often not known how these features behave in comparison with true formants. The purpose of this study was to compare two sets of automatically extracted formant-like features, i.e., robust formants and HMM2 features, to hand-labeled formants. The robust formant features were derived by means of the split Levinson algorithm while the HMM2 features correspond to the frequency segmentation of speech signals obtained by two-dimensional hidden Markov models. Mel-frequency cepstral coefficients (MFCCs) were also included in the investigation as an example of state-of-the-art automatic speech recognition features. The feature sets were compared in terms of their performance on a vowel classification task. The speech data and hand-labeled formants that were used in this study are a subset of the American English vowels database presented in Hillenbrand et al. [J. Acoust. Soc. Am. 97, 3099-3111 (1995)]. Classification performance was measured on the original, clean data and in noisy acoustic conditions. When using clean data, the classification performance of the formant-like features compared very well to the performance of the hand-labeled formants in a gender-dependent experiment, but was inferior to the hand-labeled formants in a gender-independent experiment. The results that were obtained in noisy acoustic conditions indicated that the formant-like features used in this study are not inherently noise robust. For clean and noisy data as well as for the gender-dependent and gender-independent experiments the MFCCs achieved the same or superior results as the formant features, but at the price of a much higher feature dimensionality.


Asunto(s)
Fonética , Acústica del Lenguaje , Algoritmos , Bases de Datos Factuales , Análisis Discriminante , Femenino , Humanos , Masculino , Cadenas de Markov , Modelos Biológicos , Ruido , Factores Sexuales
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