Departments of 1 Physiology, 2 Medicine (Renal
Division), and 3 Pathology, Emory University School of Medicine,
Atlanta, Georgia 30322; and 4 Department of Medicine, College of
Medicine Electron ...Microscopy Core Facility, University of Florida
College of Medicine, Gainesville, Florida 32610
A new polyclonal antibody to the human
erythrocyte urea transporter UT-B detects a broad band between 45 and
65 kDa in human erythrocytes and between 37 and 51 kDa in rat
erythrocytes. In human erythrocytes, the UT-B protein is the Kidd (Jk)
antigen, and Jk(a+b+) erythrocytes express the 45- to 65-kDa band.
However, in Jk null erythrocytes Jk(a b ), only a faint band at
55 kDa is detected. In kidney medulla, a broad band between 41 and 54 kDa, as well as a larger band at 98 kDa, is detected. Human and rat
kidney show UT-B staining in nonfenestrated endothelial cells in
descending vasa recta. UT-B protein and mRNA are detected in rat brain,
colon, heart, liver, lung, and testis. When kidney medulla or liver
proteins are analyzed with the use of a native gel, only a single
protein band is detected. UT-B protein is detected in cultured bovine
endothelial cells. We conclude that UT-B protein is expressed in more
rat tissues than previously reported, as well as in erythrocytes.
kidney; brain; liver; testis; lung; heart; colon; Kidd antigen; endothelial cells
*
R. T. Timmer and J. D. Klein
contributed equally to this work.
We have isolated and characterized the human homolog of the rat largest urea transporter of the UT-A family (hUT-A1). The 4.2-kb hUT-A1 cDNA encodes a 920-amino acid peptide, which is 89% identical ...to the rat UT-A1 protein. By Northern hybridization, hUT-A1 expression is detected in the human inner medulla as a approximately 4.4-kb mRNA transcript. By Western analysis, hUT-A1 is identified as a approximately 100-kDa protein in the human inner medulla. By immunohistochemistry, hUT-A1 expression is localized to the inner medullary collecting duct (IMCD). When transfected into HEK-293 cells hUT-A1 cDNA is translated into a approximately 98-kDa protein. Expression of hUT-A1 in Xenopus oocytes results in phloretin-inhibitable uptake of (14)C-urea, which shows only modest stimulation by cAMP, suggesting that in the human IMCD vasopressin may have a limited role in the short-term regulation of hUT-A1-mediated urea transport. We determined the organization of the human Slc14a2 gene and identified 20 exons distributed over approximately 67.5 kb on chromosome 18, from which hUT-A1 and the other human urea transporter, hUT-A2, are transcribed.
With the startup of LHC, the ALICE detector will collect data at a rate that, after two years, will reach 4PB per year. To process such a large amount of data, ALICE has developed AliEn, a ...distributed computing environment, integrated with the WLCG environment. The ALICE environment presents several original solutions, which have shown their viability in a number of large exercises of increasing complexity called ALICE Data Challenges. Within the ALICE distributed computing environment, the AliEn Workload Management Structure was created to submit to the WLCG infrastructure, and has played a crucial role to achieve the mentioned results. ALICE has more than 80 sites distributed all over the world and this WMS together with the operations management structure defined by the experiment has demonstrated a reliability and performance level ready to begin the data taking at the end of the year. In this talk we will focus on the description and current status of the AliEn WMS, emphasizing the last functionalities that have been included to handle from a single entry point the different matchmaking services of WLCG (lcg-RB, gLite WMS) and also the CREAM Computing Element; the latter has been extensively tested by the experiment during summer 2008.
The implementation of a database of digitised mammograms is discussed. The digitised images were collected beginning in 1999 by a community of physicists in collaboration with radiologists in several ...Italian hospitals as a first step in developing and implementing a computer-aided detection (CAD) system. All 3,369 mammograms were collected from 967 patients and classified according to lesion type and morphology, breast tissue and pathology type. A dedicated graphical user interface was developed to visualise and process mammograms to support the medical diagnosis directly on a high-resolution screen. The database has been the starting point for developing other medical imaging applications, such as a breast CAD, currently being upgraded and optimised for use in a distributed environment with grid services, in the framework of the Instituto Nazionale di Fisicia Nucleare (INFN)-funded Medical Applications on a Grid Infrastructure Connection (MAGIC)-5 project.
ABSTRACT
We describe a search for gravitational waves from compact binaries with at least one component with mass $0.2$–$1.0 \, \mathrm{M}_\odot$ and mass ratio q ≥ 0.1 in Advanced Laser ...Interferometer Gravitational-Wave Observatory (LIGO) and Advanced Virgo data collected between 2019 November 1, 15:00 utc and 2020 March 27, 17:00 utc. No signals were detected. The most significant candidate has a false alarm rate of $0.2 \, \rm {yr}^{-1}$. We estimate the sensitivity of our search over the entirety of Advanced LIGO’s and Advanced Virgo’s third observing run, and present the most stringent limits to date on the merger rate of binary black holes with at least one subsolar-mass component. We use the upper limits to constrain two fiducial scenarios that could produce subsolar-mass black holes: primordial black holes (PBH) and a model of dissipative dark matter. The PBH model uses recent prescriptions for the merger rate of PBH binaries that include a rate suppression factor to effectively account for PBH early binary disruptions. If the PBHs are monochromatically distributed, we can exclude a dark matter fraction in PBHs $f_\mathrm{PBH} \gtrsim \, 0.6$ (at 90 per cent confidence) in the probed subsolar-mass range. However, if we allow for broad PBH mass distributions, we are unable to rule out fPBH = 1. For the dissipative model, where the dark matter has chemistry that allows a small fraction to cool and collapse into black holes, we find an upper bound fDBH < 10−5 on the fraction of atomic dark matter collapsed into black holes.
Heterogeneity in disease course and treatment response among patients with MCD/FSGS necessitates a granular evaluation of kidney tissue features. This study aimed to identify histologic and ...ultrastructural descriptors of structural changes most predictive of clinical outcomes in the Nephrotic Syndrome Study Network (NEPTUNE).
Forty-eight histologic (37 glomerular, 9 tubulointerstitial, 2 vascular) and 20 ultrastructural descriptors were quantified by applying the NEPTUNE Digital Pathology Scoring System to NEPTUNE kidney biopsies. Outcomes included time from biopsy to disease progression, first complete remission of proteinuria, and treatment response. Relative importance of pathology and clinical predictors was obtained from random forest models, and predictive discrimination was assessed.
Among 224 participants (34% Black, 24% Hispanic), model performance was excellent, with predictive discrimination of 0.9 for disease progression, 0.85 for complete remission, and 0.81 for treatment response. The most predictive descriptors of outcomes included both conventional-
, global sclerosis or segmental sclerosis and interstitial fibrosis/tubular atrophy-and novel features, including adhesion, interstitial foam cells, deflation, periglomerular fibrosis, mononuclear white blood cells, endothelial cell abnormalities, microvillous transformation, and acute tubular injury.
The most predictive descriptors of clinical outcomes among MCD/FSGS patients reflected structural changes in multiple renal compartments. Reporting these descriptors should be standardized to guide prognostication of proteinuric glomerular diseases.
Aldose reductase aldehyde reductase 2; alditol:NAD(P)+1-oxidoreductase, EC 1.1.1.21 catalyzes conversion of glucose to sorbitol. Although its activity is implicated in the progression of ocular and ...neurological complications of diabetes, the normal function of the enzyme in most cells is unknown. Both aldose reductase activity and substantial levels of sorbitol were previously reported in renal inner medullary cells. In this tissue, the extracellular NaCl concentration normally is high and varies considerably depending on the urine concentration. We report here on a line of renal medullary cells in which medium that is high in NaCl greatly increases both aldose reductase activity and intracellular sorbitol. In these tissue culture cells (and presumably also in the renal inner medulla), the intracellular sorbitol helps balance the osmotic pressure of elevated extracellular NaCl and thus prevents cellular dehydration.
The mechanism that concentrates the urine to an osmolality several times that of systemic plasma results in high concentrations of solutes (particularly NaCl and urea) in extracellular fluid of renal ...medulla, but not in the labyrinth of the renal cortex. Intracellular and extracellular osmolality must be equal in animals, but the known intracellular levels of Na and K salts and urea in renal medullas are much too low to balance the high extracellular osmolality. The purpose of these studies was to identify the other intracellular osmolytes that must be present. Cortexes and medullas from rabbit and rat kidneys were analyzed by proton nuclear magnetic resonance, mass spectrometry, and chemical assays to determine the identity and amount of organic solutes. Large amounts of glycerophosphorylcholine, betaine, sorbitol, and inositol were found in both species localized almost exclusively to the inner medulla. In rabbits during antidiuresis glycerophosphorylcholine, betaine, and sorbitol were present in the inner medulla, at concentrations of 21.1, 34.8, and 20.8 mumol/g wet weight, respectively, but were not detected in the cortex. Inositol was present in rabbit inner medulla at 10.7 mumol/g wet weight and was also present in the cortex, but at lower concentration. None of the above metabolites was present in measurable amounts in urine or peripheral plasma. The accumulation in the cells of the inner medulla of relatively large amounts of betaine, sorbitol, glycerophosphorylcholine and inositol during antidiuresis suggests that they may play a significant role in the maintenance of intracellular osmotic balance.
Purpose
In mass spectrometry (MS)‐based studies to discover urinary protein biomarkers, an important question is how to analyze the data to find the most promising potential biomarkers to be advanced ...to large‐scale validation studies. Here, we describe a “systems biology‐based” approach to address this question.
Experimental design
We analyzed large‐scale liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) data of urinary exosomes from renal allograft recipients with biopsy‐proven evidence of immunological rejection or tubular injury (TI). We asked whether bioinformatic analysis of urinary exosomal proteins can identify biological‐process based protein groups that correlate with biopsy findings and whether the protein groups fit with general knowledge of the pathophysiological mechanisms involved.
Results
LC‐MS/MS analysis of urinary exosomal proteomes identified more than 1000 proteins in each pathologic group. These protein lists were analyzed computationally to identify the Biological Process and Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway terms that are significantly associated with each pathological group. Among the most informative terms for each group were: “sodium ion transport” for TI; “immune response” for all rejection; “epithelial cell differentiation” for cell‐mediated rejection; and “acute inflammatory response” for antibody‐mediated rejection. Based on these terms, candidate biomarkers were identified using a novel strategy to allow a dichotomous classification between different pathologic categories.
Conclusions and clinical relevance
The terms and candidate biomarkers identified make rational connections to pathophysiological mechanisms, suggesting that the described bioinformatic approach will be useful in advancing large‐scale biomarker identification studies toward a validation phase.
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