El laboratorio Vondriska es apoyado por subvenciones del Instituto Nacional del Corazón, los Pulmones y la Sangre del NIH y la Fundación Laubisch de la UCLA. EM es beneficiaria de la Jennifer S. Buchwald de Becas de Postgrado en Fisiología de la UCLA; HC es el receptor de una Asociación Americana del Corazón Pre-doctoral Fellowship, MP es el destinatario de un NIH Ruth Kirschstein beca posdoctoral, y SF es el receptor de un NIH K99 Award.

<ol>
	<li>Li, F., Wang, X., Capasso, J. M., Gerdes, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+transition+of+cardiac+myocytes+from+hyperplasia+to+hypertrophy+during+postnatal+development.">Rapid transition of cardiac myocytes from hyperplasia to hypertrophy during postnatal development.</a> J Mol Cell Cardiol. 28, 1737-1746 (1996).</li><li>Rumyantsev, P. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interrelations+of+the+proliferation+and+differentiation+processes+during+cardiact+myogenesis+and+regeneration.">Interrelations of the proliferation and differentiation processes during cardiact myogenesis and regeneration.</a> Int Rev. Cytol. 51, 186-273 (1977).</li><li>Olson, E. N., Schneider, M. 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L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+disease+and+stroke+statistics--2011+update:+a+report+from+the+American+Heart+Association.">Heart disease and stroke statistics--2011 update: a report from the American Heart Association.</a> Circulation. 123, e18-e209 (2011).</li><li>Schirmer, E. C., Florens, L., Guan, T., Yates, J. R., Gerace, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nuclear+membrane+proteins+with+potential+disease+links+found+by+subtractive+proteomics.">Nuclear membrane proteins with potential disease links found by subtractive proteomics.</a> Science. 301, 1380-1382 (2003).</li><li>Lambert, J. 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K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Post-translational+regulation+of+calsarcin-1+during+pressure+overload-induced+cardiac+hypertrophy.">Post-translational regulation of calsarcin-1 during pressure overload-induced cardiac hypertrophy.</a> Journal of molecular and cellular cardiology. 48, 1206-1214 (2010).</li><li>Franklin, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+analysis+of+the+chromatin+proteome+in+disease+reveals+remodeling+principles+and+identifies+high+mobility+group+protein+b2+as+a+regulator+of+hypertrophic+growth.">Quantitative analysis of the chromatin proteome in disease reveals remodeling principles and identifies high mobility group protein b2 as a regulator of hypertrophic growth.</a> Mol Cell Proteomics. 11, (2012).</li><li>Gramolini, A. O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+proteomics+profiling+of+a+phospholamban+mutant+mouse+model+of+dilated+cardiomyopathy+reveals+progressive+intracellular+stress+responses.">Comparative proteomics profiling of a phospholamban mutant mouse model of dilated cardiomyopathy reveals progressive intracellular stress responses.</a> Mol Cell Proteomics. 7, 519-533 (2008).</li><li>Singh, H., Warburton, S., Vondriska, T. M., Khakh, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proteomics+to+Identify+Proteins+Interacting+with+P2X2+Ligand-Gated+Cation+Channels.">Proteomics to Identify Proteins Interacting with P2X2 Ligand-Gated Cation Channels.</a> J. Vis. Exp. (27), e1178 (2009).</li><li>Park, S. K., Venable, J. D., Xu, T., Yates, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+quantitative+analysis+software+tool+for+mass+spectrometry-based+proteomics.">A quantitative analysis software tool for mass spectrometry-based proteomics.</a> Nat. Methods. 5, 319-322 (2008).</li><li>Lomenick, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Target+identification+using+drug+affinity+responsive+target+stability+(DARTS).">Target identification using drug affinity responsive target stability (DARTS).</a> Proc. Natl. Acad. Sci. U. S. A. 106, 21984-21989 (2009).</li><li>Kamleh, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolomic+profiling+using+Orbitrap+Fourier+transform+mass+spectrometry+with+hydrophilic+interaction+chromatography:+a+method+with+wide+applicability+to+analysis+of+biomolecules.">Metabolomic profiling using Orbitrap Fourier transform mass spectrometry with hydrophilic interaction chromatography: a method with wide applicability to analysis of biomolecules.</a> Rapid Commun. Mass Spectrom. 22, 1912-1918 (2008).</li><li>Searle, B. 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J. 77, 631-636 (1960).</li><li>Rodriguez-Collazo, P., Leuba, S. H., Zlatanova, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Robust+methods+for+purification+of+histones+from+cultured+mammalian+cells+with+the+preservation+of+their+native+modifications.">Robust methods for purification of histones from cultured mammalian cells with the preservation of their native modifications.</a> Nucleic Acids Res. 37, e81 (2009).</li><li>Kuehl, L., Salmond, B., Tran, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Concentrations+of+high-mobility-group+proteins+in+the+nucleus+and+cytoplasm+of+several+rat+tissues.">Concentrations of high-mobility-group proteins in the nucleus and cytoplasm of several rat tissues.</a> J. Cell Biol. 99, 648-654 (1984).</li><li>Gronow, M., Griffiths, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+isolation+and+separation+of+the+non-histone+proteins+of+rat+liver+nuclei.">Rapid isolation and separation of the non-histone proteins of rat liver nuclei.</a> FEBS Lett. 15, 340-344 (1971).</li><li>Mischke, B. G., Ward, O. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+tissue+specificity+of+chromatin-associated+proteins+in+Vicia+faba.">Isolation and tissue specificity of chromatin-associated proteins in Vicia faba.</a> Can J. Biochem. 53, 91-95 (1975).</li><li>Andersen, J. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Directed+proteomic+analysis+of+the+human+nucleolus.">Directed proteomic analysis of the human nucleolus.</a> Curr. Biol. 12, 1-11 (2002).</li><li>Zhao, Y., Jensen, O. 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No hay conflictos de interés declarado.

Existen dos métodos principales para el aislamiento nuclear han sido revisados ​​previamente: 27 una es la técnica Behrens de homogeneizar tejido liofilizado en un disolvente no acuoso y la segunda, una modificación de la que se utiliza aquí, de homogenizar el tejido en un acuosa de sacarosa / solución salina seguido por diferencial o centrifugación en gradiente de densidad. Subfraccionamiento de los núcleos mediante extracción con ácido en muestras de tejido es una h...

<table border="1"><tbody><tr><td> Nombre del reactivo </td><td> Empresa </td><td> Número de catálogo </td></tr><tr><td> Dulbeco Modificado Medio Eagle </td><td> Invitrogen </td><td> 11965 </td></tr><tr><td> Proteasa pellet </td><td> Roche </td><td> 04 693 159 001 </td></tr><tr><td> 100 m colador </td><td> BD Falcon </td><td> 352360 </td></tr><tr><td> Ultracut ultramicrótomo </td><td> Reichert </td><td></td></tr><tr><td> 100CX Electrónica de Transmisión Microscopio </td><td> JEOL EE.UU., Inc. </td><td></td></tr><tr><td> Oriol </td><td> BioRad </td><td> 161-0496 </td></tr><tr><td> Histona H2A anticuerpo </td><td> Santa Cruz </td><td> sc-8648 </td></tr><tr><td> Nucleoporin p62 anticuerpo </td><td> BD Biosciences </td><td> 610498 </td></tr><tr><td> Adenina nucleomarea anticuerpo transportador </td><td> Santa Cruz </td><td> sc-9299 </td></tr><tr><td> Bip anticuerpo </td><td> Santa Cruz </td><td> sc-1050 </td></tr><tr><td> Tubulina de anticuerpos </td><td> Sigma </td><td> T1568 </td></tr><tr><td> Histona H3 anticuerpo </td><td> Abcam </td><td> ab1791 </td></tr><tr><td> Fibrilarina anticuerpo </td><td> Señalización Celular </td><td> C12C3 </td></tr><tr><td> SNRP70 anticuerpo </td><td> Abcam </td><td> ab51266 </td></tr><tr><td> E2F-1 anticuerpo </td><td> Thermo Fisher </td><td> MS-879 </td></tr><tr><td> Retinoblastoma anticuerpo </td><td> BD Biosciences </td><td> 554136 </td></tr><tr><td> Factor-1 inducible por hipoxia anticuerpo </td><td> Novus Productos Biológicos </td><td> NB100-469 </td></tr><tr><td> BCA ensayo de proteínas </td><td> Thermo Scientific </td><td> 23227 </td></tr><tr><td> Columna de fase inversa </td><td> Obj Nuevoective </td><td> PFC7515-B14-10 </td></tr><tr><td> BioWorks Browser </td><td> Thermo Scientific </td><td></td></tr></tbody></table>

quantitative analysis of chromatin proteomes in disease

En el núcleo residen los proteomas cuyas funciones están íntimamente vinculadas con la regulación de genes. Núcleos de cardiomiocitos adultos de mamíferos son únicos debido al alto porcentaje de células binucleadas, un estado predominantemente heterochromatic del ADN, y la naturaleza no división de los cardiomiocitos que hace que los núcleos adulto en un estado permanente de interfase. 2 Regulación transcripcional durante el desarrollo y enfermedad han sido bien estudiados en este órgano, 3-5, pero lo que permanece relativamente inexplorado es el papel que desempeñan las proteínas nucleares responsables de empaquetamiento del ADN y la expresión, y cómo estas proteínas controlan los cambios en los programas transcripcionales que ocurren durante la enfermedad. 6 En los países desarrollados mundo, la enfermedad cardíaca es la principal causa de mortalidad tanto en hombres como en mujeres. 7 Insight sobre cómo las proteínas nucleares cooperar para regular la progresión de esta enfermedad es fundamental para avanzar en el tratamiento actual opciones. La espectrometría de masas es la herramienta ideal para abordar estas cuestiones, ya que permite una anotación imparcial de la cuantificación nuclear proteoma y relativo de cómo la abundancia de estas proteínas cambia con la enfermedad. Si bien se han realizado varios estudios proteómicos para mamíferos complejos de proteínas nucleares, 8-13 hasta hace 14 sólo ha habido un estudio que examina el proteoma nuclear cardíaca, y se considera el núcleo entero, en lugar de explorar el proteoma a nivel de sub-compartimentos nucleares . 15 En gran parte, esta escasez de trabajo se debe a la dificultad de aislar núcleos cardíaco. Núcleos cardiacos ocurren dentro de un rígido y denso de actina-miosina aparato al que están conectados a través de múltiples extensiones desde el retículo endoplásmico, en la medida en que altera la contracción de miocitos su forma general. 16 Además, los cardiomiocitos son 40% mitocondrias en volumen 17 que necessitàtes enriquecimiento del núcleo aparte de los otros orgánulos. Aquí se describe un protocolo para el enriquecimiento nuclear cardíaca y fraccionamiento adicional en compartimentos biológicamente relevantes. Además, los métodos de detalle de la etiqueta libre de disección de espectrometría de masa cuantitativa de estas fracciones-técnicas susceptibles de experimentación in vivo en varios modelos animales y sistemas de órganos en el etiquetado metabólico no es factible.

La figura 4 pone de manifiesto la utilidad de esta forma de cuantificación relativa. Se muestra en el panel de la izquierda son los picos individuales monoisotópicas péptidos (superpuesta de diferentes ratones), que han sido designados como pertenecientes a la proteína HMGB1 (identificados mediante una búsqueda de base de datos). Cada pico, esencialmente un cromatógrafo iónico extrae para el péptido dado, proviene de un ratón diferente. Los grupos representan tres diferentes estados fisiológic...

Watch this Scientific Journal Video about Quantitative Analysis of Chromatin Proteomes in Disease at JoVE.com

Quantitative Analysis of Chromatin Proteomes in Disease

Los avances en la espectrometría de masas han permitido el análisis de alto rendimiento de la expresión de proteínas y la modificación en un huésped de tejidos. En combinación con el fraccionamiento subcelular y modelos de enfermedad, espectrometría de masa cuantitativa y bioinformática puede revelar nuevas propiedades en los sistemas biológicos. El método descrito en la presente memoria analiza asociadas a la cromatina proteínas en el contexto de las enfermedades del corazón y es fácilmente aplicable a otras<em&gt; In vivo</em&gt; Modelos de enfermedad humana.

Análisis cuantitativo de proteomas cromatina en la enfermedad

In the nucleus reside the proteomes whose functions are most intimately linked with gene regulation. Adult mammalian cardiomyocyte nuclei are unique due ...

quantitative-analysis-of-chromatin-proteomes-in-disease

Research

JoVE Journal

Medicine

13.0K vistas.  David Geffen School of Medicine at UCLA. Los avances en la espectrometría de masas han permitido el análisis de alto rendimiento de la expresión de proteínas y la modificación en un huésped de tejidos. En combinación con el fraccionamiento subcelular y modelos de enfermedad, espectrometría de masa cuantitativa y bioinformática puede revelar nuevas propiedades en los sistemas biológicos. El método descrito en la presente memoria analiza asociadas a la cromatina proteínas en el contexto de las enfermed...

Video: Quantitative Analysis of Chromatin Proteomes in Disease

Quantitative Analysis of Cancer Metastasis using an Avian Embryo Model

During metastasis cancer cells disseminate from the primary tumor, invade into surrounding tissues, and spread to distant organs. Metastasis is a complex process that can involve many tissue types, span variable time periods, and often occur deep within organs, making it difficult to investigate and quantify. In addition, the efficacy of the metastatic process is influenced by multiple steps in the metastatic cascade making it difficult to evaluate the contribution of a single aspect of tumor cell behavior. As a consequence, metastasis assays are frequently performed in experimental animals to provide a necessarily realistic context in which to study metastasis. Unfortunately, these models are further complicated by their complex physiology. The chick embryo is a unique in vivo model that overcomes many limitations to studying metastasis, due to the accessibility of the chorioallantoic membrane (CAM), a well-vascularized extra-embryonic tissue located underneath the eggshell that is receptive to the xenografting of tumor cells (figure 1). Moreover, since the chick embryo is naturally immunodeficient, the CAM readily supports the engraftment of both normal and tumor tissues. Most importantly, the avian CAM successfully supports most cancer cell characteristics including growth, invasion, angiogenesis, and remodeling of the microenvironment. This makes the model exceptionally useful for the investigation of the pathways that lead to cancer metastasis and to predict the response of metastatic cancer to new potential therapeutics. The detection of disseminated cells by species-specific Alu PCR makes it possible to quantitatively assess metastasis in organs that are colonized by as few as 25 cells. Using the Human Epidermoid Carcinoma cell line (HEp3) we use this model to analyze spontaneous metastasis of cancer cells to distant organs, including the chick liver and lung. Furthermore, using the Alu-PCR protocol we demonstrate the sensitivity and reproducibility of the assay as a tool to analyze and quantitate intravasation, arrest, extravasation, and colonization as individual elements of metastasis.

Using quantitative PCR, we demonstrate how the well-established chick CAM model can be used to quantitatively analyze the metastasis of human tumor cells to distant organs.

Análisis cuantitativo de la metástasis del cáncer utilizando un modelo de embrión aviar

During metastasis cancer cells disseminate from the primary tumor, invade into surrounding tissues, and spread to distant organs. Metastasis is a complex ...

Investigación

Medicina

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

Un sencillo método de intubación ratón de pulmón

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have been developed to facilitate the study of the molecular and cellular pathophysiology of this disease. In this manuscript we describe specific techniques for the quantification and characterization of atherosclerotic lesions in the murine aortic sinus and ascending aorta. The advantage of this procedure is that it provides an accurate measurement of the cross-sectional area and total volume of the lesion, which can be used to compare atherosclerotic progression across different treatment groups. This is possible through the use of the valve leaflets as an anatomical landmark, together with careful adjustment of the sectioning angle. We also describe basic staining methods that can be used to begin to characterize atherosclerotic progression. These can be further modified to investigate antigens of specific interest to the researcher. The described techniques are generally applicable to a wide variety of existing and newly created dietary and genetically-induced models of atherogenesis.

We describe procedures to quantify and characterize atherosclerotic lesions in mouse models using precision sectioning of the aortic sinus and ascending aorta combined with histochemical and immunohistochemical analysis.

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Quantitative Magnetic Resonance Imaging of Skeletal Muscle Disease

Quantitative magnetic resonance imaging (qMRI) describes the development and use of MRI to quantify physical, chemical, and/or biological properties of living systems. Neuromuscular diseases often exhibit a temporally varying, spatially heterogeneous, and multi-faceted pathology. The goal of this protocol is to characterize this pathology using qMRI methods. The MRI acquisition protocol begins with localizer images (used to locate the position of the body and tissue of interest within the MRI system), quality control measurements of relevant magnetic field distributions, and structural imaging for general anatomical characterization. The qMRI portion of the protocol includes measurements of the longitudinal and transverse relaxation time constants (T1 and T2, respectively). Also acquired are diffusion-tensor MRI data, in which water diffusivity is measured and used to infer pathological processes such as edema. Quantitative magnetization transfer imaging is used to characterize the relative tissue content of macromolecular and free water protons. Lastly, fat-water MRI methods are used to characterize fibro-adipose tissue replacement of muscle. In addition to describing the data acquisition and analysis procedures, this paper also discusses the potential problems associated with these methods, the analysis and interpretation of the data, MRI safety, and strategies for artifact reduction and protocol optimization.

Neuromuscular diseases often exhibit a temporally varying, spatially heterogeneous, and multi-faceted pathology. The goal of this protocol is to characterize this pathology using non-invasive magnetic resonance imaging methods.

Imágenes por resonancia magnética cuantitativa de la enfermedad del músculo esquelético

Quantitative magnetic resonance imaging (qMRI) describes the development and use of MRI to quantify physical, chemical, and/or biological properties of ...

Quantitative Mass Spectrometric Profiling of Cancer-cell Proteomes Derived From Liquid and Solid Tumors

In-depth analyses of cancer cell proteomes are needed to elucidate oncogenic pathomechanisms, as well as to identify potential drug targets and diagnostic biomarkers. However, methods for quantitative proteomic characterization of patient-derived tumors and in particular their cellular subpopulations are largely lacking. Here we describe an experimental set-up that allows quantitative analysis of proteomes of cancer cell subpopulations derived from either liquid or solid tumors. This is achieved by combining cellular enrichment strategies with quantitative Super-SILAC-based mass spectrometry followed by bioinformatic data analysis. To enrich specific cellular subsets, liquid tumors are first immunophenotyped by flow cytometry followed by FACS-sorting; for solid tumors, laser-capture microdissection is used to purify specific cellular subpopulations. In a second step, proteins are extracted from the purified cells and subsequently combined with a tumor-specific, SILAC-labeled spike-in standard that enables protein quantification. The resulting protein mixture is subjected to either gel electrophoresis or Filter Aided Sample Preparation (FASP) followed by tryptic digestion. Finally, tryptic peptides are analyzed using a hybrid quadrupole-orbitrap mass spectrometer, and the data obtained are processed with bioinformatic software suites including MaxQuant. By means of the workflow presented here, up to 8,000 proteins can be identified and quantified in patient-derived samples, and the resulting protein expression profiles can be compared among patients to identify diagnostic proteomic signatures or potential drug targets.

In-depth analyses of cancer cell proteomes facilitate identification of novel drug targets and diagnostic biomarkers. We describe an experimental workflow for quantitative analysis of (phospho-)proteomes in cancer cell subpopulations derived from liquid and solid tumors. This is achieved by combining cellular enrichment strategies with quantitative Super-SILAC-based mass spectrometry.

Misa cuantitativa espectrometría de perfiles de los proteomas de células del cáncer derivados de líquidos y tumores sólidos

In-depth analyses of cancer cell proteomes are needed to elucidate oncogenic pathomechanisms, as well as to identify potential drug targets and diagnostic ...

Quantitative 3D In Silico Modeling (q3DISM) of Cerebral Amyloid-beta Phagocytosis in Rodent Models of Alzheimer's Disease

Neuroinflammation is now recognized as a major etiological factor in neurodegenerative disease. Mononuclear phagocytes are innate immune cells responsible for phagocytosis and clearance of debris and detritus. These cells include CNS-resident macrophages known as microglia, and mononuclear phagocytes infiltrating from the periphery. Light microscopy has generally been used to visualize phagocytosis in rodent or human brain specimens. However, qualitative methods have not provided definitive evidence of in vivo phagocytosis. Here, we describe quantitative 3D in silico modeling (q3DISM), a robust method allowing for true 3D quantitation of amyloid-&#946; (A&#946;) phagocytosis by mononuclear phagocytes in rodent Alzheimer&#39;s Disease (AD) models. The method involves fluorescently visualizing A&#946; encapsulated within phagolysosomes in rodent brain sections. Large z-dimensional confocal datasets are then 3D reconstructed for quantitation of A&#946; spatially colocalized within the phagolysosome. We demonstrate the successful application of q3DISM to mouse and rat brains, but this methodology can be extended to virtually any phagocytic event in any tissue.

Quantitative 3D In Silico Modeling q3DISM of Cerebral Amyloid-beta Phagocytosis in Rodent Models of Alzheimer&#039;s Disease

We developed a methodology for quantitative 3D in silico modeling (q3DISM) of cerebral amyloid-&#946; (A&#946;) phagocytosis by mononuclear phagocytes in rodent models of Alzheimer&#39;s disease. This method can be generalized for the quantitation of virtually any phagocytic event in vivo.

3D cuantitativa<em&gt; In silico</em&gt; Modelado (q3DISM) de beta-amiloide cerebral fagocitosis en roedores modelos de enfermedad de Alzheimer

Neuroinflammation is now recognized as a major etiological factor in neurodegenerative disease. Mononuclear phagocytes are innate immune cells responsible ...

Application of Granger Causality Analysis of the Directed Functional Connection in Alzheimer's Disease and Mild Cognitive Impairment

Impaired functional connectivity in the Default Mode Network (DMN) may be involved in the progression of Alzheimer&#39;s Disease (AD). The Posterior Cingulate Cortex (PCC) is a potential imaging marker for monitoring the progression of AD. Previous studies did not focus on the functional connectivity between the PCC and nodes in regions outside the DMN, but our study is an effort to explore these overlooked functional connections. For collecting data, we used functional Magnetic Resonance Imaging (fMRI) and Granger Causality Analysis (GCA). fMRI provides a non-invasive method for studying the dynamic interactions between the different brain regions. GCA is a statistical hypothesis test for determining whether one-time series is useful in forecasting another. In simple terms, it is judged by comparing the &#34;Known all the information on the last moment, the distribution of the probability of X at this time&#34; and the &#34;Known all the information on the last moment except Y, the distribution of the probability of X at this time&#34;, to determine whether there is a causal relationship between Y and X. This definition is based on the complete information source and stationary chronological sequence. The main step of this analysis is to use X and Y to establish the regression equation and draw a causal relationship by a hypothetical test. Since GCA can measure causal effects, we used it to investigate the anisotropy of the functional connectivity and explore the hub function of the PCC. Here, we screened 116 participants for MRI scanning, and after preprocessing the data obtained from neuroimaging, we used GCA to derive the causal relationship of each node. Finally, we concluded that the directed connection is significantly different between the Mild Cognitive Impairment (MCI) and AD groups, both from the PCC to the whole brain and from the whole brain to the PCC.

Application of Granger Causality Analysis of the Directed Functional Connection in Alzheimer&#039;s Disease and Mild Cognitive Impairment

Based on resting-state functional magnetic resonance imaging with Granger causality analysis, we investigated the alterations in the directed functional connectivity between the posterior cingulate cortex and whole brain in patients with Alzheimer&#39;s Disease (AD), patients with Mild Cognitive Impairment (MCI), and healthy controls.

Aplicación del análisis de causalidad de Granger de la conexión funcional dirigida en la enfermedad de Alzheimer y deterioro cognitivo leve

Impaired functional connectivity in the Default Mode Network (DMN) may be involved in the progression of Alzheimer&#39;s Disease (AD). The Posterior ...

In Vivo Morphometric Analysis of Human Cranial Nerves Using Magnetic Resonance Imaging in Meni&#232;re's Disease Ears and Normal Hearing Ears

Analysis of neural structures in Meni&#232;re's Disease (MD) is of importance, since a loss of such structures has previously been proposed for this patient group but has yet to be confirmed. This protocol describes a method of in vivo evaluation of neural changes especially well suitable for cranial nerve analysis using magnetic resonance imaging (MRI). MD-patients and normal hearing persons were examined in a 3-T MR-scanner using a scan protocol including strongly T2-weighted 3D gradient-echo-sequence (3D-CISS). In the patient group, MD was additionally confirmed using MRI-based assessment of endolymphatic hydrops. Morphometric analysis was performed using a freeware DICOM viewer. Evaluation of cranial nerves included measurements of cross-sectional areas (CSAs) of the nerves at different levels as well as orthogonal diametric measurements.

&lt;em&gt;In Vivo&lt;/em&gt; Morphometric Analysis of Human Cranial Nerves Using Magnetic Resonance Imaging in Meni&amp;#232;re&#039;s Disease Ears and Normal Hearing Ears

To evaluate morphological changes of cranial nerves such as loss of neural structures or swelling of cranial nerves in Meni&#232;re&#39;s Disease (MD) or in healthy persons in vivo, a protocol of evaluation has been developed using magnetic resonance imaging (MRI). Additional MRI-based confirmation of MD was performed.

En Vivo Análisis morfométrico de los nervios craneales de humanos usando resonancia magnética en la enfermedad orejas y oídos de la audiencia Normal de Menière

Analysis of neural structures in Meni&#232;re's Disease (MD) is of importance, since a loss of such structures has previously been proposed for this ...

Whole-brain Segmentation and Change-point Analysis of Anatomical Brain MRI&#8212;Application in Premanifest Huntington's Disease

Recent advances in MRI offer a variety of useful markers to identify neurodegenerative diseases. In Huntington&#39;s disease (HD), regional brain atrophy begins many years prior to the motor onset (during the &#34;premanifest&#34; period), but the spatiotemporal pattern of regional atrophy across the brain has not been fully characterized. Here we demonstrate an online cloud-computing platform, &#34;MRICloud&#34;, which provides atlas-based whole-brain segmentation of T1-weighted images at multiple granularity levels, and thereby, enables us to access the regional features of brain anatomy. We then describe a regression model that detects statistically significant inflection points, at which regional brain atrophy starts to be noticeable, i.e. the &#34;change-point&#34;, with respect to a disease progression index. We used the CAG-age product (CAP) score to index the disease progression in HD patients. Change-point analysis of the volumetric measurements from the segmentation pipeline, therefore, provides important information of the order and pattern of structural atrophy across the brain. The paper illustrates the use of these techniques on T1-weighted MRI data of premanifest HD subjects from a large multicenter PREDICT-HD study. This design potentially has wide applications in a range of neurodegenerative diseases to investigate the dynamic changes of brain anatomy.

Whole-brain Segmentation and Change-point Analysis of Anatomical Brain MRI&amp;#8212;Application in Premanifest Huntington&#039;s Disease

This paper describes a statistical model for volumetric MRI data analysis, which identifies the &#34;change-point&#34; when brain atrophy begins in premanifest Huntington&#39;s disease. Whole-brain mapping of the change-points is achieved based on brain volumes obtained using an atlas-based segmentation pipeline of T1-weighted images.

Todo cerebro segmentación y análisis de punto de cambio anatómico cerebral MRI — aplicación asintomático enfermedad de Huntington

Recent advances in MRI offer a variety of useful markers to identify neurodegenerative diseases. In Huntington&#39;s disease (HD), regional brain atrophy ...

Quantitative [18F]-Naf-PET-MRI Analysis for the Evaluation of Dynamic Bone Turnover in a Patient with Facetogenic Low Back Pain

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. Bone is a dynamic tissue undergoing continuous remodeling with the competing activity of osteoblasts, which produce the new bone matrix, and osteoclasts, whose function is to eliminate mineralized bone. [18F]-NaF is a positron emission tomography (PET) radiotracer that enables visualization of bone metabolism. [18F]-NaF is chemically absorbed into hydroxyapatite in the bone matrix by osteoblasts and can thus noninvasively detect osteoblastic activity, which is occult to conventional imaging techniques. Kinetic modeling of dynamic [18F]-NaF-PET data provides detailed quantitative measures of bone metabolism. Conventional semi-quantitative PET data, which utilizes standardized uptake values (SUVs) as a measure of radiotracer activity, is referred to as a static technique due to its snapshot of tracer uptake in time.&#160; Kinetic modeling, however, utilizes dynamic image data where tracer levels are continuously acquired providing tracer uptake temporal resolution. From the kinetic modeling of dynamic data, quantitative values like blood flow and metabolic rate (i.e., potentially informative metrics of tracer dynamics) can be extracted, all with respect to the measured activity in the image data. When combined with dual modality PET-MRI, region-specific kinetic data can be correlated with anatomically registered high-resolution structural and pathologic information afforded by MRI. The goal of this methodological manuscript is to outline detailed techniques for performing and analyzing dynamic [18F]-NaF-PET-MRI data. The lumbar facet joint is a common site of degenerative arthritis disease and a common cause for axial low back pain.&#160; Recent studies suggest [18F]-NaF-PET may serve as a useful biomarker of painful facetogenic disease.&#160; The human lumbar facet joint will, therefore, be used as a prototypical region of interest for dynamic [18F]-NaF-PET-MRI analysis in this manuscript.

Quantitative [18F]-Naf-PET-MRI Analysis for the Evaluation of Dynamic Bone Turnover in a Patient with Facetogenic Low Back Pain

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. We present detailed methodologies for performing and analyzing dynamic [18F]-NaF-PET-MRI data in a patient with facetogenic low back pain using the lumbar facet joints as a prototypical region of interest.

Cuantitativo [18F]-Naf-PET-MRI Análisis para la evaluación de la rotación ósea dinámica en un paciente con dolor de espalda baja facetogénico

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. Bone is a dynamic tissue undergoing ...