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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science, Ltd
    Journal of neuroendocrinology 13 (2001), S. 0 
    ISSN: 1365-2826
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Melatonin acts via high affinity, G-protein coupled, seven transmembrane domain receptors. To precisely localize these receptors, antibodies were raised in chickens against a 15 amino acid fragment at the intracellular C-terminal region of the human melatonin receptor subtype mt1 (DSSNDVADRVKWKPS, mt1338−352). A chimeric form of the receptor with a hydrophilic Flag peptide (DYKDDDDK) in sequence with the extracellular N-terminus (Flag-mt1) was generated by polymerase chain reaction and expressed in mammalian cell lines. An IgY antibody (Y31), which gave high antibody titres by enzyme-linked immunosorbent assay, was used to localize Flag-mt1 in stably transfected cells by immunofluoresence. Flag-mt1 localization with Y31 was identical to that obtained with the M5 antibody directed against the Flag epitope and was mainly localized to the Golgi apparatus with some staining at the cell surface. No staining was seen in untransfected cells with either antibody. Y31 staining was abolished using antibody preabsorbed with peptide antigen. Y31 immunofluorescence in fetal human kidney sections was restricted to nephrogenic regions and matched that of 2-(125I)iodomelatonin binding and mt1 gene expression by in situ hybridization. Y31 was used to immunoprecipitate biotinylated membrane proteins from Flag-mt1 stably transfected and untransfected CHO cells. Western blotting of immunoprecipitated proteins revealed two major bands specific to stably transfected cells, one at 63 kDa and one at 86 kDa. The first band almost certainly corresponds to the glycosylated form of Flag-mt1 and the second band to receptor dimers. Thus, Y31 antibody is suitable for use in detecting the human mt1 receptor subtype in tissues and in transfected cells.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-2932
    Keywords: tropospheric ozone ; white clover ; air pollution ; biomonitor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract A white clover (Trifolium repens L.) system using measured biomass to indicate effective concentrations of tropospheric ozone (O3) has been developed. The system utilizes the relative response of an O3-sensitive clone (NC-S) and an O3-resistant clone (NC-R) grown in 15-liter pots. Forage (leaves, stems and flowers) is cut, dried, and weighed at 28-day intervals. Forage dry weight ratios (NC-S/NC-R) for individual or multiple harvests indicate O3 concentrations during growth. In, 3 years of testing in open-top field chambers at Raleigh, North Carolina, O3 always decreased growth of NC-S more than that of NC-R and the NC-S/NC-R ratio routinely decreased as the O3 concentration increased. A national field test was performed in 1993 and 1994 to determine if the clover system can account for effects of climatic variables on clover growth per-se, and if climatic variables affect the relative response of the two clones to O3. Eight locations (Corvallis, Oregon; Kennedy Space Center, Florida; Delaware, Ohio; Amherst, Massachusetts; Blacksburg, Virginia; Raleigh, North Carolina; Riverside, California; San Bemardino mountains, California) provided large differences in O3 concentrations and climate. The NC-S/NC-R forage ratios for three consecutive 28-day growth periods for each year as related to the mean 12 hour per day O3 concentrations are presented in this manuscript. Ratios were generally highest where mean O3 concentrations were lowest (Oregon and Florida), lowest where mean O3 concentrations were highest (both California locations), and intermediate at other locations.
    Type of Medium: Electronic Resource
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  • 3
    Publication Date: 2013-06-02
    Description: Chronic pancreatitis (CP) is a devastating disease characterized by persistent and uncontrolled abdominal pain. Our lack of understanding is partially due to the lack of experimental models that mimic the human disease and also to the lack of validated behavioral measures of visceral pain. The ligand-gated cation channel transient receptor potential ankyrin 1 (TRPA1) mediates inflammation and pain in early experimental pancreatitis. It is unknown if TRPA1 causes fibrosis and sustained pancreatic pain. We induced CP by injecting the chemical agent trinitrobenzene sulfonic acid (TNBS), which causes severe acute pancreatitis, into the pancreatic duct of C57BL/6 trpa1 +/+ and trpa1 -/- mice. Chronic inflammatory changes and pain behaviors were assessed after 2–3 wk. TNBS injection caused marked pancreatic fibrosis with increased collagen-staining intensity, atrophy, fatty replacement, monocyte infiltration, and pancreatic stellate cell activation, and these changes were reflected by increased histological damage scores. TNBS-injected animals showed mechanical hypersensitivity during von Frey filament probing of the abdomen, decreased daily voluntary wheel-running activity, and increased immobility scores during open-field testing. Pancreatic TNBS also reduced the threshold to hindpaw withdrawal to von Frey filament probing, suggesting central sensitization. Inflammatory changes and pain indexes were significantly reduced in trpa1 -/- mice. In conclusion, we have characterized in mice a model of CP that resembles the human condition, with marked histological changes and behavioral measures of pain. We have demonstrated, using novel and objective pain measurements, that TRPA1 mediates inflammation and visceral hypersensitivity in CP and could be a therapeutic target for the treatment of sustained inflammatory abdominal pain.
    Print ISSN: 0193-1857
    Electronic ISSN: 1522-1547
    Topics: Medicine
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  • 4
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    The Society of Nuclear Medicine (SNM)
    Publication Date: 2016-02-02
    Description: Gastrointestinal bleeding scintigraphy performed with 99m Tc-labeled autologous erythrocytes or historically with 99m Tc-sulfur colloid has been a clinically useful tool since the 1970s. This article reviews the history of the techniques, the different methods of radiolabeling erythrocytes, the procedure, useful indications, diagnostic accuracy, the use of SPECT/CT and CT angiography to evaluate gastrointestinal bleeding, and Meckel diverticulum imaging. The causes of pediatric bleeding are discussed by age.
    Print ISSN: 0022-3123
    Topics: Medicine
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  • 5
    Publication Date: 2014-02-04
    Description: Although iodinated radiopharmaceuticals usually contain a small quantity of unbound iodine, it is difficult to establish the degree to which thyroid activity on scintigraphic images reflects uptake of free radioiodine. The objective of the present study was to examine the effectiveness of thyroid blockade in subjects undergoing 123 I- meta -iodobenzylguanidine ( m IBG) imaging and to estimate the relative contribution of bound and unbound radioiodine to imaging findings. Methods: All subjects were participants in prospective trials of 123 I- m IBG cardiac imaging in which pretreatment with thyroid blockade was optional unless locally required. In a pilot project, 15 subjects (6 blocked) had thyroid uptake measured at 4 h using a probe system. Fifteen-minute (early) and 4-h (late) anterior planar chest images that included the thyroid region were visually scored for thyroid uptake (scale of 0–4) in another group of 152 subjects (98 blocked). Quantitative analysis based on thyroid regions of interest was performed on anterior planar images from a further sample of 669 subjects (442 blocked). For all 3 investigations, quantitative comparisons of thyroid uptake were made between the blocked and nonblocked subjects. Results: There was no statistical difference between probe uptake of the 6 blocked and 9 nonblocked subjects. However, in the second series, mean visual score on the late images was significantly lower for blocked than nonblocked subjects ( P 〈 0.001). In the region-of-interest analyses, net thyroid counts were significantly higher on the late images of nonblocked subjects ( P 〈 0.0001), and compared with early images, 87% of subjects who received blockade showed decreased or unchanged counts whereas 75% of nonblocked subjects had increased net thyroid activity. In nonblocked subjects, an estimated 79% of thyroid counts on late images could be attributed to unbound 123 I. Conclusion: On the basis of 3 different methods for assessing thyroid uptake of 123 I, use of thyroid blockade pretreatment in 123 I- m IBG imaging prevents increase of thyroid activity over time because of uptake of unbound 123 I. In most subjects, there is a low level of 123 I- m IBG thyroid activity that probably represents specific uptake in sympathetic nerve terminals.
    Print ISSN: 0022-3123
    Topics: Medicine
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  • 6
    Publication Date: 2017-11-02
    Print ISSN: 0022-3123
    Topics: Medicine
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  • 7
    Publication Date: 2012-10-16
    Description: Cathepsins regulate premature trypsinogen activation within acinar cells, a key initial step in pancreatitis. The identity, origin, and causative roles of activated cathepsins in pancreatic inflammation and pain are not defined. By using a near infrared-labeled activity-based probe (GB123) that covalently modifies active cathepsins, we localized and identified activated cathepsins in mice with cerulein-induced pancreatitis and in pancreatic juice from patients with chronic pancreatitis. We used inhibitors of activated cathepsins to define their causative role in pancreatic inflammation and pain. After GB123 administration to mice with pancreatitis, reflectance and confocal imaging showed significant accumulation of the probe in inflamed pancreas compared with controls, particularly in acinar cells and macrophages, and in spinal cord microglia and neurons. Biochemical analysis of pancreatic extracts identified them as cathepsins B, L, and S (Cat-B, Cat-L, and Cat-S, respectively). These active cathepsins were also identified in pancreatic juice from patients with chronic pancreatitis undergoing an endoscopic procedure for the treatment of pain, indicating cathepsin secretion. The cathepsin inhibitor K11777 suppressed cerulein-induced activation of Cat-B, Cat-L, and Cat-S in the pancreas and ameliorated pancreatic inflammation, nocifensive behavior, and activation of spinal nociceptive neurons. Thus pancreatitis is associated with an increase in the active forms of the proteases Cat-B, Cat-L, and Cat-S in pancreatic acinar cells and macrophages, and in spinal neurons and microglial cells. Inhibition of cathepsin activation ameliorated pancreatic inflammation and pain. Activity-based probes permit identification of proteases that are predictive biomarkers of disease progression and response to therapy and may be useful noninvasive tools for the detection of pancreatic inflammation.
    Print ISSN: 0193-1857
    Electronic ISSN: 1522-1547
    Topics: Medicine
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  • 8
    Publication Date: 2013-01-09
    Description: Enteritis caused by Clostridium difficile toxin (Tx) is a nosocomial disease of increasing clinical concern, but the local mediators of C. difficile TxA inflammation are unknown. The potent vasodilator calcitonin gene-related peptide mediates neurogenic inflammation via the calcitonin receptor-like receptor (CLR). Here we examined the ileum-specific effects of reducing CLR...
    Print ISSN: 0027-8424
    Electronic ISSN: 1091-6490
    Topics: Biology , Medicine , Natural Sciences in General
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  • 9
  • 10
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    Unknown
    The Society of Nuclear Medicine (SNM)
    Publication Date: 2012-11-02
    Print ISSN: 0022-3123
    Topics: Medicine
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