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1.
Psychol Res ; 2024 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-38940823

RESUMO

Human beings are able to imagine actions with the aim to change movement coordination and to learn particular movements. Meta-analyses to date have shown that when individuals systematically engage in imagery of a motor action without overt behavior this can improve motor performance and facilitate motor learning. Despite a considerable body of research in neuroscience, psychology, and sport science, however, there is at present no consensus on the neurocognitive mechanisms of imagery, and the mechanisms that lead to learning via imagined action are still being debated. In particular, the differences between imagined and overt action, and respective learning effects, remain to be fully explained. The present collection of manuscripts is a result of compiling both theoretical advances in the field of motor control and motor learning and those in imagery research to better understand imagery and learning. It is structured alongside five position papers from leading experts in the field, each of which is followed by a series of short commentaries written by experts from various disciplines. This collection demonstrates (a) that conceptualizations of imagery are manifold, vary highly and depend on the perspective chosen, (b) that existing approaches to the neurocognitive mechanisms of imagery and imagery practice of motor actions draw on distinct motor control and learning perspectives, (c) that perspectives from the wider fields of motor control and learning stimulate new approaches to explain imagery and imagery practice, (d) and that future research is needed to investigate and compare different perspectives and conceptualizations of the neurocognitive mechanisms of imagery and imagery practice of motor actions.

2.
Pflege ; 37(2): 89-97, 2024 Apr.
Artigo em Alemão | MEDLINE | ID: mdl-37997948

RESUMO

The use of the DOS and Delirium Prevalence: a quantitative longitudinal study at a Swiss-German central hospital Abstract: Background: With a prevalence of 12-64%, delirium is a common complication in acute care, associated with negative outcomes such as increased mortality and prolonged length of stay. Many hospitals have guidelines to improve the delirium management. The Delirium Observation Screening Scale (DOS) Score is collected in the study hospital from all patients ≥ 70 years at each shift for at least 3 days. Delirium is diagnosed by a physician and coded according to ICD-10. Purpose: Evaluation of the delirium screening with the DOS according to internal guideline in terms of number of DOS assessments performed, prevalence of delirium (DOS score ≥ 3 points, CD-10 code delirium). Method: This retrospective quantitative single-centre longitudinal study used 2017 and 2018 data of 10046 cases. Statistical analysis methods were used to analyse prevalence of delirium and subgroup comparisons. Results: At least one DOS score was documented in 92% of cases aged ≥ 70-years (n = 5038). DOS implementation varied between 60% in the early, 49% in the late and 38% in the night shift. The prevalence of delirium was 12% according to DOS score ≥ 3 and 4% according to physician diagnosis of a delirium. Cases with a DOS score ≥ 3 were significantly older, more often female, had more comorbidities and were depressed. Conclusions: DOS is performed in most patients when indicated. The DOS implementation frequency varied depending on the shift.


Assuntos
Delírio , Feminino , Humanos , Delírio/diagnóstico , Delírio/epidemiologia , Hospitais , Estudos Longitudinais , Estudos Retrospectivos , Suíça , Masculino , Idoso
3.
Biol Reprod ; 107(2): 406-418, 2022 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-35466369

RESUMO

Zinc influx and efflux events are essential for meiotic progression in oocytes of several mammalian and amphibian species, but it is less clear whether this evolutionary conservation of zinc signals is also important in late-stage germline development in invertebrates. Using quantitative, single cell elemental mapping methods, we find that Caenorhabditis elegans oocytes undergo significant stage-dependent fluctuations in total zinc content, rising by over sevenfold from Prophase I through the beginning of mitotic divisions in the embryo. Live imaging of the rapid cell cycle progression in C. elegans enables us to follow changes in labile zinc pools across meiosis and mitosis in single embryo. We find a dynamic increase in labile zinc prior to fertilization that then decreases from Anaphase II through pronuclear fusion and relocalizes to the eggshell. Disruption of these zinc fluxes blocks extrusion of the second polar body, leading to a range of mitotic defects. We conclude that spatial temporal zinc fluxes are necessary for meiotic progression in C. elegans and are a conserved feature of germ cell development in a broad cross section of metazoa.


Assuntos
Caenorhabditis elegans , Zinco , Animais , Caenorhabditis elegans/genética , Caenorhabditis elegans/metabolismo , Fertilização , Mamíferos/metabolismo , Meiose , Oócitos/metabolismo , Zinco/metabolismo
4.
Opt Express ; 30(15): 26027-26042, 2022 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-36236801

RESUMO

As a coherent diffraction imaging technique, ptychography provides high-spatial resolution beyond Rayleigh's criterion of the focusing optics, but it is also sensitively affected by the decoherence coming from the spatial and temporal variations in the experiment. Here we show that high-speed ptychographic data acquisition with short exposure can effectively reduce the impact from experimental variations. To reach a cumulative dose required for a given resolution, we further demonstrate that a continuous multi-pass scan via high-speed ptychography can achieve high-resolution imaging. This low-dose scan strategy is shown to be more dose-efficient, and has potential for radiation-sensitive sample studies and time-resolved imaging.

5.
Nature ; 532(7600): 496-9, 2016 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-26982721

RESUMO

Problematic fossils, extinct taxa of enigmatic morphology that cannot be assigned to a known major group, were once a major issue in palaeontology. A long-favoured solution to the 'problem of the problematica', particularly the 'weird wonders' of the Cambrian Burgess Shale, was to consider them representatives of extinct phyla. A combination of new evidence and modern approaches to phylogenetic analysis has now resolved the affinities of most of these forms. Perhaps the most notable exception is Tullimonstrum gregarium, popularly known as the Tully monster, a large soft-bodied organism from the late Carboniferous Mazon Creek biota (approximately 309-307 million years ago) of Illinois, USA, which was designated the official state fossil of Illinois in 1989. Its phylogenetic position has remained uncertain and it has been compared with nemerteans, polychaetes, gastropods, conodonts, and the stem arthropod Opabinia. Here we review the morphology of Tullimonstrum based on an analysis of more than 1,200 specimens. We find that the anterior proboscis ends in a buccal apparatus containing teeth, the eyes project laterally on a long rigid bar, and the elongate segmented body bears a caudal fin with dorsal and ventral lobes. We describe new evidence for a notochord, cartilaginous arcualia, gill pouches, articulations within the proboscis, and multiple tooth rows adjacent to the mouth. This combination of characters, supported by phylogenetic analysis, identifies Tullimonstrum as a vertebrate, and places it on the stem lineage to lampreys (Petromyzontida). In addition to increasing the known morphological disparity of extinct lampreys, a chordate affinity for T. gregarium resolves the nature of a soft-bodied fossil which has been debated for more than 50 years.


Assuntos
Fósseis , Filogenia , Vertebrados/classificação , Nadadeiras de Animais/anatomia & histologia , Animais , Extinção Biológica , Olho/anatomia & histologia , Trato Gastrointestinal/anatomia & histologia , Illinois , Lampreias/classificação , Notocorda/anatomia & histologia , Dente/anatomia & histologia , Vertebrados/anatomia & histologia
6.
Psychol Res ; 2022 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-36574019

RESUMO

In this paper, we discuss a variety of ways in which practising motor actions by means of motor imagery (MI) can be enhanced via synchronous action observation (AO), that is, by AO + MI. We review the available research on the (mostly facilitatory) behavioural effects of AO + MI practice in the early stages of skill acquisition, discuss possible theoretical explanations, and consider several issues related to the choice and presentation schedules of suitable models. We then discuss considerations related to AO + MI practice at advanced skill levels, including expertise effects, practical recommendations such as focussing attention on specific aspects of the observed action, using just-ahead models, and possible effects of the perspective in which the observed action is presented. In section "Coordinative AO + MI", we consider scenarios where the observer imagines performing an action that complements or responds to the observed action, as a promising and yet under-researched application of AO + MI training. In section "The dual action simulation hypothesis of AO + MI", we review the neurocognitive hypothesis that AO + MI practice involves two parallel action simulations, and we consider opportunities for future research based on recent neuroimaging work on parallel motor representations. In section "AO + MI training in motor rehabilitation", we review applications of AO, MI, and AO + MI training in the field of neurorehabilitation. Taken together, this evidence-based, exploratory review opens a variety of avenues for future research and applications of AO + MI practice, highlighting several clear advantages over the approaches of purely AO- or MI-based practice.

7.
J Synchrotron Radiat ; 28(Pt 1): 309-317, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33399582

RESUMO

Ptychography is a rapidly developing scanning microscopy which is able to view the internal structures of samples at a high resolution beyond the illumination size. The achieved spatial resolution is theoretically dose-limited. A broadband source can provide much higher flux compared with a monochromatic source; however, it conflicts with the necessary coherence requirements of this coherent diffraction imaging technique. In this paper, a multi-wavelength reconstruction algorithm has been developed to deal with the broad bandwidth in ptychography. Compared with the latest development of mixed-state reconstruction approach, this multi-wavelength approach is more accurate in the physical model, and also considers the spot size variation as a function of energy due to the chromatic focusing optics. Therefore, this method has been proved in both simulation and experiment to significantly improve the reconstruction when the source bandwidth, illumination size and scan step size increase. It is worth mentioning that the accurate and detailed information of the energy spectrum for the incident beam is not required in advance for the proposed method. Further, we combine multi-wavelength and mixed-state approaches to jointly solve temporal and spatial partial coherence in ptychography so that it can handle various disadvantageous experimental effects. The significant relaxation in coherence requirements by our approaches allows the use of high-flux broadband X-ray sources for high-efficient and high-resolution ptychographic imaging.

8.
Nucleic Acids Res ; 47(22): 11807-11825, 2019 12 16.
Artigo em Inglês | MEDLINE | ID: mdl-31722427

RESUMO

Modifications of ribosomal RNA expand the nucleotide repertoire and thereby contribute to ribosome heterogeneity and translational regulation of gene expression. One particular m5C modification of 25S ribosomal RNA, which is introduced by Rcm1p, was previously shown to modulate stress responses and lifespan in yeast and other small organisms. Here, we report that NSUN5 is the functional orthologue of Rcm1p, introducing m5C3782 into human and m5C3438 into mouse 28S ribosomal RNA. Haploinsufficiency of the NSUN5 gene in fibroblasts from William Beuren syndrome patients causes partial loss of this modification. The N-terminal domain of NSUN5 is required for targeting to nucleoli, while two evolutionary highly conserved cysteines mediate catalysis. Phenotypic consequences of NSUN5 deficiency in mammalian cells include decreased proliferation and size, which can be attributed to a reduction in total protein synthesis by altered ribosomes. Strikingly, Nsun5 knockout in mice causes decreased body weight and lean mass without alterations in food intake, as well as a trend towards reduced protein synthesis in several tissues. Together, our findings emphasize the importance of single RNA modifications for ribosome function and normal cellular and organismal physiology.


Assuntos
Crescimento e Desenvolvimento/genética , Metiltransferases/genética , Proteínas Musculares/genética , Biossíntese de Proteínas/genética , Animais , Peso Corporal/genética , Crescimento Celular , Proliferação de Células/genética , Células Cultivadas , Criança , Embrião de Mamíferos , Feminino , Deleção de Genes , Células HEK293 , Células HeLa , Humanos , Lactente , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout
9.
Pflege ; 34(6): 329-336, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34445884

RESUMO

No chance for pneumonia - A campaign for mobilization in the context of a practice project addressing pneumonia prevention Abstract. Background: Healthcare-associated infections (HAI) in inpatients are associated with complicated treatment. In Europe, 5.5 % of inpatients develop HAI. About half of these infections could be avoided. In the Clinic for traumatology of the university hospital Zurich, we developed interventions to reduce HAI. Thereby, we focused on non-ventilator-associated hospital-acquired pneumonia (nvHAP). Aim: Besides reducing nvHAP rates, we intended to improve patient mobility, to empower nurses, and to strengthen interprofessional collaboration. Methods: To achieve these aims, we performed a practice development project comprising inhouse training, workshops, information posters, structural changes and a mobilization campaign. Results: Patient mobilization increased by 40 %, duration of mobilization sessions by 46.5 %. The semi-annual comparison shows a sustainable improvement of 7.6 %. Nurses reported knowledge gain, considerably improved interprofessional collaboration and increased quality of caring. Discussion: Combining various methods and following an interprofessional approach resulted in sustainable effects. Limitations and transfer: Targeted practice development proves to be suitable for promoting patient mobility. Regular repetitions and physiotherapy services at off-peak times are essential to ensure sustainability.


Assuntos
Infecção Hospitalar , Hospitais Universitários , Humanos , Pacientes Internados
10.
Opt Express ; 28(11): 16407-16416, 2020 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-32549464

RESUMO

We present an interrogation laser system for a transportable strontium lattice clock operating at 698 nm, which is based on an ultra-low-expansion glass reference cavity. Transportability is achieved by implementing a rigid, compact, and vibration insensitive mounting of the 12 cm-long reference cavity, sustaining shocks of up to 50 g. The cavity is mounted at optimized support points that independently constrain all degrees of freedom. This mounting concept is especially beneficial for cavities with a ratio of length L over diameter DL/D > 1. Generally, large L helps to reduce thermal noise-induced laser frequency instability while small D leads to small cavity volume. The frequency instability was evaluated, reaching its thermal noise floor of mod σy ≈ 3 × 10-16 for averaging times between 0.5 s and 10 s. The laser system was successfully operated during several field studies.

11.
Biotechnol Bioeng ; 117(4): 1037-1047, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31956981

RESUMO

The encapsulation of biopharmaceuticals into micro- or nanoparticles is a strategy frequently used to prevent degradation or to achieve the slow release of therapeutics and vaccines. Protein bodies (PBs), which occur naturally as storage organelles in seeds, can be used as such carrier vehicles. The fusion of the N-terminal sequence of the maize storage protein, γ-zein, to other proteins is sufficient to induce the formation of PBs, which can be used to bioencapsulate recombinant proteins directly in the plant production host. In addition, the immunostimulatory effects of zein have been reported, which are advantageous for vaccine delivery. However, little is known about the interaction between zein PBs and mammalian cells. To better understand this interaction, fluorescent PBs, resulting from the fusion of the N-terminal portion of zein to a green fluorescent protein, was produced in Nicotiana benthamiana leaves, recovered by a filtration-based downstream procedure, and used to investigate their internalization efficiency into mammalian cells. We show that fluorescent PBs were efficiently internalized into intestinal epithelial cells and antigen-presenting cells (APCs) at a higher rate than polystyrene beads of comparable size. Furthermore, we observed that PBs stimulated cytokine secretion by epithelial cells, a characteristic that may confer vaccine adjuvant activities through the recruitment of APCs. Taken together, these results support the use of zein fusion proteins in developing novel approaches for drug delivery based on controlled protein packaging into plant PBs.


Assuntos
Produtos Biológicos , Proteínas de Fluorescência Verde , Proteínas Recombinantes de Fusão , Zeína , Administração Oral , Produtos Biológicos/administração & dosagem , Produtos Biológicos/química , Produtos Biológicos/metabolismo , Linhagem Celular , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Folhas de Planta/metabolismo , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Nicotiana/genética , Nicotiana/metabolismo , Células U937 , Zeína/química , Zeína/genética , Zeína/metabolismo
12.
Arterioscler Thromb Vasc Biol ; 39(11): 2320-2337, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31554420

RESUMO

OBJECTIVE: Copper (Cu) is essential micronutrient, and its dysregulation is implicated in aortic aneurysm (AA) development. The Cu exporter ATP7A (copper-transporting P-type ATPase/Menkes ATPase) delivers Cu via the Cu chaperone Atox1 (antioxidant 1) to secretory Cu enzymes, such as lysyl oxidase, and excludes excess Cu. Lysyl oxidase is shown to protect against AA formation. However, the role and mechanism of ATP7A in AA pathogenesis remain unknown. Approach and Results: Here, we show that Cu chelator markedly inhibited Ang II (angiotensin II)-induced abdominal AA (AAA) in which ATP7A expression was markedly downregulated. Transgenic ATP7A overexpression prevented Ang II-induced AAA formation. Conversely, Cu transport dysfunctional ATP7Amut/+/ApoE-/- mice exhibited robust AAA formation and dissection, excess aortic Cu accumulation as assessed by X-ray fluorescence microscopy, and reduced lysyl oxidase activity. In contrast, AAA formation was not observed in Atox1-/-/ApoE-/- mice, suggesting that decreased lysyl oxidase activity, which depends on both ATP7A and Atox1, was not sufficient to develop AAA. Bone marrow transplantation suggested importance of ATP7A in vascular cells, not bone marrow cells, in AAA development. MicroRNA (miR) array identified miR-125b as a highly upregulated miR in AAA from ATP7Amut/+/ApoE-/- mice. Furthermore, miR-125b target genes (histone methyltransferase Suv39h1 and the NF-κB negative regulator TNFAIP3 [tumor necrosis factor alpha induced protein 3]) were downregulated, which resulted in increased proinflammatory cytokine expression, aortic macrophage recruitment, MMP (matrix metalloproteinase)-2/9 activity, elastin fragmentation, and vascular smooth muscle cell loss in ATP7Amut/+/ApoE-/- mice and reversed by locked nucleic acid-anti-miR-125b infusion. CONCLUSIONS: ATP7A downregulation/dysfunction promotes AAA formation via upregulating miR-125b, which augments proinflammatory signaling in a Cu-dependent manner. Thus, ATP7A is a potential therapeutic target for inflammatory vascular disease.


Assuntos
Aneurisma da Aorta Abdominal/genética , Aneurisma da Aorta Abdominal/fisiopatologia , ATPases Transportadoras de Cobre/fisiologia , MicroRNAs/fisiologia , Angiotensina II/efeitos dos fármacos , Animais , Apoptose , Células Cultivadas , Quelantes/farmacologia , Cobre/metabolismo , Proteínas de Transporte de Cobre/metabolismo , ATPases Transportadoras de Cobre/genética , Modelos Animais de Doenças , Regulação para Baixo , Feminino , Humanos , Inflamação/genética , Inflamação/fisiopatologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Chaperonas Moleculares/metabolismo , Molibdênio/farmacologia , Músculo Liso Vascular/citologia , Regulação para Cima
13.
Opt Lett ; 44(17): 4331-4334, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31465395

RESUMO

As x-ray microscopy is pushed into the nanoscale with the advent of more bright and coherent x-ray sources, associated improvement in spatial resolution becomes highly vulnerable to geometrical errors and uncertainties during data collection. We address a form of error in tomography experiments, namely, the drift between projections during the tomographic scan. Our proposed method can simultaneously recover the drift, while tomographically reconstructing the specimen based on a joint iterative optimization scheme. This approach utilizes the correlation provided from different view angles and different signals. While generally applicable, we demonstrate our method on x-ray fluorescence tomography from a tissue specimen and compare the reconstruction quality with conventional methods.

14.
Cereb Cortex ; 28(3): 907-923, 2018 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-28077513

RESUMO

Imitation learning involves the acquisition of novel motor patterns based on action observation (AO). We used event-related functional magnetic resonance imaging to study the imitation learning of spatial sequences and rhythms during AO, motor imagery (MI), and imitative execution in nonmusicians and musicians. While both tasks engaged the fronto-parietal mirror circuit, the spatial sequence task recruited posterior parietal and dorsal premotor regions more strongly. The rhythm task involved an additional network for auditory working memory. This partial dissociation supports the concept of task-specific mirror mechanisms. Two regions of cognitive control were identified: 1) dorsolateral prefrontal cortex (DLPFC) was found to be more strongly activated during MI of novel spatial sequences, which allowed us to extend the 2-level model of imitation learning by Buccino et al. (2004) to spatial sequences. 2) During imitative execution of both tasks, the posterior medial frontal cortex was robustly activated, along with the DLPFC, which suggests that both regions are involved in the cognitive control of imitation learning. The musicians' selective behavioral advantage for rhythm imitation was reflected cortically in enhanced sensory-motor processing during AO and by the absence of practice-related activation differences in DLPFC during rhythm execution.


Assuntos
Córtex Cerebral/diagnóstico por imagem , Cognição/fisiologia , Comportamento Imitativo/fisiologia , Imageamento por Ressonância Magnética , Periodicidade , Aprendizagem Espacial/fisiologia , Estimulação Acústica , Adolescente , Adulto , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Memória de Curto Prazo/fisiologia , Destreza Motora/fisiologia , Música , Rede Nervosa/diagnóstico por imagem , Oxigênio/sangue , Adulto Jovem
15.
Proc Natl Acad Sci U S A ; 112(8): 2314-9, 2015 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-25675478

RESUMO

Trace metals play important roles in normal and in disease-causing biological functions. X-ray fluorescence microscopy reveals trace elements with no dependence on binding affinities (unlike with visible light fluorophores) and with improved sensitivity relative to electron probes. However, X-ray fluorescence is not very sensitive for showing the light elements that comprise the majority of cellular material. Here we show that X-ray ptychography can be combined with fluorescence to image both cellular structure and trace element distribution in frozen-hydrated cells at cryogenic temperatures, with high structural and chemical fidelity. Ptychographic reconstruction algorithms deliver phase and absorption contrast images at a resolution beyond that of the illuminating lens or beam size. Using 5.2-keV X-rays, we have obtained sub-30-nm resolution structural images and ∼90-nm-resolution fluorescence images of several elements in frozen-hydrated green algae. This combined approach offers a way to study the role of trace elements in their structural context.


Assuntos
Clorófitas/anatomia & histologia , Congelamento , Imageamento Tridimensional/métodos , Difração de Raios X/métodos , Clorófitas/citologia , Microscopia de Fluorescência
16.
Behav Res Methods ; 50(5): 2004-2015, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29076105

RESUMO

Nowadays, the use of eyetracking to determine 2-D gaze positions is common practice, and several approaches to the detection of 2-D fixations exist, but ready-to-use algorithms to determine eye movements in three dimensions are still missing. Here we present a dispersion-based algorithm with an ellipsoidal bounding volume that estimates 3D fixations. Therefore, 3D gaze points are obtained using a vector-based approach and are further processed with our algorithm. To evaluate the accuracy of our method, we performed experimental studies with real and virtual stimuli. We obtained good congruence between stimulus position and both the 3D gaze points and the 3D fixation locations within the tested range of 200-600 mm. The mean deviation of the 3D fixations from the stimulus positions was 17 mm for the real as well as for the virtual stimuli, with larger variances at increasing stimulus distances. The described algorithms are implemented in two dynamic linked libraries (Gaze3D.dll and Fixation3D.dll), and we provide a graphical user interface (Gaze3DFixGUI.exe) that is designed for importing 2-D binocular eyetracking data and calculating both 3D gaze points and 3D fixations using the libraries. The Gaze3DFix toolkit, including both libraries and the graphical user interface, is available as open-source software at https://github.com/applied-cognition-research/Gaze3DFix .


Assuntos
Algoritmos , Fixação Ocular/fisiologia , Imageamento Tridimensional/instrumentação , Software , Adulto , Feminino , Humanos , Imageamento Tridimensional/métodos , Masculino , Pessoa de Meia-Idade
17.
J Synchrotron Radiat ; 24(Pt 1): 288-295, 2017 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-28009569

RESUMO

The study of a multilayered and multicomponent system by spatially resolved X-ray fluorescence microscopy poses unique challenges in achieving accurate quantification of elemental distributions. This is particularly true for the quantification of materials with high X-ray attenuation coefficients, depth-dependent composition variations and thickness variations. A widely applicable procedure for use after spectrum fitting and quantification is described. This procedure corrects the elemental distribution from the measured fluorescence signal, taking into account attenuation of the incident beam and generated fluorescence from multiple layers, and accounts for sample thickness variations. Deriving from Beer-Lambert's law, formulae are presented in a general integral form and numerically applicable framework. The procedure is applied using experimental data from a solar cell with a Cu(In,Ga)Se2 absorber layer, measured at two separate synchrotron beamlines with varied measurement geometries. This example shows the importance of these corrections in real material systems, which can change the interpretation of the measured distributions dramatically.

18.
J Microsc ; 265(1): 81-93, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27580164

RESUMO

Trace metals play important roles in biological function, and x-ray fluorescence microscopy (XFM) provides a way to quantitatively image their distribution within cells. The faithfulness of these measurements is dependent on proper sample preparation. Using mouse embryonic fibroblast NIH/3T3 cells as an example, we compare various approaches to the preparation of adherent mammalian cells for XFM imaging under ambient temperature. Direct side-by-side comparison shows that plunge-freezing-based cryoimmobilization provides more faithful preservation than conventional chemical fixation for most biologically important elements including P, S, Cl, K, Fe, Cu, Zn and possibly Ca in adherent mammalian cells. Although cells rinsed with fresh media had a great deal of extracellular background signal for Cl and Ca, this approach maintained cells at the best possible physiological status before rapid freezing and it does not interfere with XFM analysis of other elements. If chemical fixation has to be chosen, the combination of 3% paraformaldehyde and 1.5 % glutaraldehyde preserves S, Fe, Cu and Zn better than either fixative alone. When chemically fixed cells were subjected to a variety of dehydration processes, air drying was proved to be more suitable than other drying methods such as graded ethanol dehydration and freeze drying. This first detailed comparison for x-ray fluorescence microscopy shows how detailed quantitative conclusions can be affected by the choice of cell preparation method.


Assuntos
Fibroblastos/química , Fibroblastos/citologia , Microscopia de Fluorescência/métodos , Espectrometria por Raios X/métodos , Fixação de Tecidos/métodos , Oligoelementos/análise , Animais , Camundongos , Células NIH 3T3
19.
Plant Biotechnol J ; 14(10): 1998-2009, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-26929151

RESUMO

Conversion of nongrain biomass into liquid fuel is a sustainable approach to energy demands as global population increases. Previously, we showed that iron can act as a catalyst to enhance the degradation of lignocellulosic biomass for biofuel production. However, direct addition of iron catalysts to biomass pretreatment is diffusion-limited, would increase the cost and complexity of biorefinery unit operations and may have deleterious environmental impacts. Here, we show a new strategy for in planta accumulation of iron throughout the volume of the cell wall where iron acts as a catalyst in the deconstruction of lignocellulosic biomass. We engineered CBM-IBP fusion polypeptides composed of a carbohydrate-binding module family 11 (CBM11) and an iron-binding peptide (IBP) for secretion into Arabidopsis and rice cell walls. CBM-IBP transformed Arabidopsis and rice plants show significant increases in iron accumulation and biomass conversion compared to respective controls. Further, CBM-IBP rice shows a 35% increase in seed iron concentration and a 40% increase in seed yield in greenhouse experiments. CBM-IBP rice potentially could be used to address iron deficiency, the most common and widespread nutritional disorder according to the World Health Organization.


Assuntos
Arabidopsis/metabolismo , Biomassa , Parede Celular/metabolismo , Ferro/metabolismo , Oryza/metabolismo , Sementes/metabolismo , Arabidopsis/genética , Biocombustíveis , Parede Celular/genética , Oryza/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
20.
Environ Sci Technol ; 50(16): 8827-39, 2016 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-27437565

RESUMO

Metal toxicity to aquatic organisms depends on the speciation of the metal and its binding to the critical receptor site(s) (biotic ligand) of the organism. The intracellular nature of the biotic ligand for Cu in microalgal cells was investigated using the high elemental sensitivity of microprobe synchrotron radiation X-ray fluorescence (SR-XRF) and X-ray absorption near-edge spectroscopy (XANES). The marine microalgae, Ceratoneis closterium, Phaeodactylum tricornutum, and Tetraselmis sp. were selected based on their varying sensitivities to Cu (72-h 50% population growth inhibitions of 8-47 µg Cu/L). Intracellular Cu in control cells was similar for all three species (2.5-3.2 × 10(-15) g Cu/cell) and increased 4-fold in C. closterium and Tetraselmis sp. when exposed to copper, but was unchanged in P. tricornutum (72-h exposure to 19, 40, and 40 µg Cu/L, respectively). Whole cell microprobe SR-XRF identified endogenous Cu in the central compartment (cytoplasm) of control (unexposed) cells. After Cu exposure, Cu was colocated with organelles/granules dense in P, S, Ca, and Si and this was clearly evident in thin sections of Tetraselmis sp. XANES indicated coexistence of Cu(I) and Cu(II) in control and Cu-exposed cells, with the Cu ligand (e.g., phytochelatin) in P. tricornutum different from that in C. closterium and Tetraselmis sp. This study supports the hypothesis that Cu(II) is reduced to Cu(I) and that polyphosphate bodies and phytochelatins play a significant role in the internalization and detoxification of Cu in marine microalgae.


Assuntos
Cobre/química , Microalgas , Diatomáceas , Síncrotrons , Raios X
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