Browsing by Author "Kamocka, Malgorzata"

Now showing 1 - 5 of 5
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    Improved Structure and Function in Autosomal Recessive Polycystic Rat Kidneys with Renal Tubular Cell Therapy
    (Public Library of Science, 2015) Kelly, Katherine J.; Zhang, Jizhong; Han, Ling; Kamocka, Malgorzata; Miller, Caroline; Gattone, Vincent H.; Dominguez, Jesus H.; Department of Medicine, IU School of Medicine
    Autosomal recessive polycystic kidney disease is a truly catastrophic monogenetic disease, causing death and end stage renal disease in neonates and children. Using PCK female rats, an orthologous model of autosomal recessive polycystic kidney disease harboring mutant Pkhd1, we tested the hypothesis that intravenous renal cell transplantation with normal Sprague Dawley male kidney cells would improve the polycystic kidney disease phenotype. Cytotherapy with renal cells expressing wild type Pkhd1 and tubulogenic serum amyloid A1 had powerful and sustained beneficial effects on renal function and structure in the polycystic kidney disease model. Donor cell engraftment and both mutant and wild type Pkhd1 were found in treated but not control PCK kidneys 15 weeks after the final cell infusion. To examine the mechanisms of global protection with a small number of transplanted cells, we tested the hypothesis that exosomes derived from normal Sprague Dawley cells can limit the cystic phenotype of PCK recipient cells. We found that renal exosomes originating from normal Sprague Dawley cells carried and transferred wild type Pkhd1 mRNA to PCK cells in vivo and in vitro and restricted cyst formation by cultured PCK cells. The results indicate that transplantation with renal cells containing wild type Pkhd1 improves renal structure and function in autosomal recessive polycystic kidney disease and may provide an intra-renal supply of normal Pkhd1 mRNA.
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    Indiana Center for Biological Microscopy
    (Office of the Vice Chancellor for Research, 2014-04-11) Kamocka, Malgorzata; Winfree, Seth; Dunn, Kenneth W.
    The Indiana Center for Biological Microscopy “ICBM” has provided imaging capabilities since 1996. The primary objective of the Center is to provide state-of-the-art microscopy facility to Indiana University researchers. In 2002 the ICBM was designated as an NIDDK George O'Brien Center for Advanced Microscopic Analysis, whose objective is to develop methods of intravital multiphoton microscopy and 3-dimentional microscopy for renal research. Since that time, the ICBM has worked with various IU investigators to develop methods for intravital microscopy of various organs. One of the recent goals of the Center is to develop an intravital microscopy service for Cancer Center investigators by establishing the Optical Microscopy Core for the CEMH. The Center is available to anyone at the Indiana University and researchers from outside the institution. We are organized around the principle of providing researchers with access to and training on the latest instruments in optical microscopy. We emphasize hands-on access to instruments for individual investigators and unparalleled assistance from on-staff imaging experts.
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    Indiana O’Brien Center for Advanced Microscopic Analysis
    (Office of the Vice Chancellor for Research, 2014-04-11) Atkinson, Simon J.; Dunn, Kenneth W.; Kamocka, Malgorzata; Molitoris, Bruce A.
    In June 2002, the IU Division of Nephrology received NIH funding to establish a George M. O'Brien Center for Advanced Renal Microscopic Analysis at the Indiana Center for Biological Microscopy. The primary goal of this Center is to develop new optical methodologies for investigators in Nephrologic and Urologic Research and to make the technology widely accessible to the renal research community. Approaches include intravital multiphoton microscopy, 3-dimensional imaging and quantitative microscopic analysis. We have established the Indiana O’Brien Fellows Program, which provides funds for investigators to implement these new techniques for their specific research needs, either in their laboratories, or utilizing the facilities of the Center.
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    Scavenger receptor class B, type I-mediated uptake of A1AT by pulmonary endothelial cells
    (American Psychological Society, 2015-08-15) Lockett, Angela D.; Petrusca, Daniela N.; Justice, Matthew J.; Porier, Christophe; Serban, Karina A.; Rush, Natalia I.; Kamocka, Malgorzata; Predescu, Dan; Predescu, Sandra; Petrache, Irina; Department of Medicine, IU School of Medicine
    In addition to exerting a potent anti-elastase function, α-1 antitrypsin (A1AT) maintains the structural integrity of the lung by inhibiting endothelial inflammation and apoptosis. A main serpin secreted in circulation by hepatocytes, A1AT requires uptake by the endothelium to achieve vasculoprotective effects. This active uptake mechanism, which is inhibited by cigarette smoking (CS), involves primarily clathrin- but also caveola-mediated endocytosis and may require active binding to a receptor. Because circulating A1AT binds to high-density lipoprotein (HDL), we hypothesized that scavenging receptors are candidates for endothelial uptake of the serpin. Although the low-density lipoprotein (LDL) receptor-related protein 1 (LRP1) internalizes only elastase-bound A1AT, the scavenger receptor B type I (SR-BI), which binds and internalizes HDL and is modulated by CS, may be involved in A1AT uptake. Transmission electron microscopy imaging of colloidal gold-labeled A1AT confirmed A1AT endocytosis in both clathrin-coated vesicles and caveolae in endothelial cells. SR-BI immunoprecipitation identified binding to A1AT at the plasma membrane. Pretreatment of human lung microvascular endothelial cells with SR-B ligands (HDL or LDL), knockdown of SCARB1 expression, or neutralizing SR-BI antibodies significantly reduced A1AT uptake by 30–50%. Scarb1 null mice exhibited decreased A1AT lung content following systemic A1AT administration and reduced lung anti-inflammatory effects of A1AT supplementation during short-term CS exposure. In turn, A1AT supplementation increased lung SR-BI expression and modulated circulating lipoprotein levels in wild-type animals. These studies indicate that SR-BI is an important mediator of A1AT endocytosis in pulmonary endothelium and suggest a cross talk between A1AT and lipoprotein regulation of vascular functions.
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    Selective Plane Illumination Microscopy, A New Imaging Modality Available at the Indiana Center for Biological Microscopy
    (Office of the Vice Chancellor for Research, 2016-04-08) Winfree, Seth; Smith, Nathaniel; Dunn, Ken; Kamocka, Malgorzata; Molitoris, Bruce
    Microscopy is a primary tool for studying 3D tissue models. Microscopy provides the only means of distinguishing the behaviors of individual cells in a heterogeneous context that obscures biochemical assays. SPIM (Selective Plane Illumination Microscopy) is new approach that is ideally suited to the unique problems involved in high-resolution imaging of 3D tissue models. In the simplest form of SPIM, a cylindrical lens is used to generate a thin lightsheet (1-10 microns) that illuminates a sample. An imaging objective lens, placed orthogonal to this lightsheet is used to collect an image of fluorescence that is selectively excited in this single illuminated plane. The sample is then rotated, and the process is repeated until a multiview dataset of the entire sample is collected. These cross-section images are then assembled to give a complete 3D image of the sample. This approach offers several advantages over conventional methods of imaging thick tissues. First, SPIM provides superior axial resolution for large field-of-view images, deconvolved SPIM volumes have isotropic 3D resolution. Second, SPIM is a “gentle” imaging approach and is better suited to imaging living tissues than either confocal or multiphoton microscopy, supporting studies of cell migration, development, signaling and physiology. Third, imaging speeds can be 30 to 200 fold faster than scanning confocal or multiphoton systems, enabling resolution of dynamic events, and rapid collection of large image datasets. We describe the assembly and customization of an OpenSPIM based lightsheet microscope (IU OpenSPIM) as a platform for developing new imaging technologies. To this end we have implemented software and hardware for multi-channel laser control and temperature and perfusion control. We present examples of highresolution, live and high speed imaging, demonstrating these capabilities. The IU OpenSPIM is a centerpiece in the development of new software for 3D tissue cytometry and a novel screening platform.