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tunel-transferase-mediated-dutp-nick-end-labeling

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Journal Articles
Journal: Diabetes
Diabetes 2007;56(2):337–345
Published: 01 February 2007
... an ability to inhibit nuclear factor-κB (NF-κB). Diabetes of 9–10 months duration significantly increased the number of TUNEL (transferase-mediated dUTP nick-end labeling)-positive capillary cells and acellular (degenerate) capillaries in the retinal vasculature, and all three salicylate-based drugs...
Journal Articles
Journal: Diabetes
Diabetes 2003;52(9):2363–2371
Published: 01 September 2003
... in L4 and L5 dorsal root ganglia of long-term experimental streptozotocin-induced diabetic rats using transferase-mediated dUTP nick-end labeling (TUNEL), 4′,6-diamidino-2-phenylindole (DAPI) staining of nuclear morphology, and electron microscopic appraisal of cell morphology. None provided any...
Journal Articles
Journal: Diabetes
Diabetes 2006;55(8):2192–2201
Published: 01 August 2006
... with adenovirus (Ad) expressing human or rat (control) proIAPP linked to green fluorescent protein, with or without Ad-PC2 or Ad-PC1/3. Expression of human proIAPP increased the number of transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells 96 h after transduction (+hIAPP 8.7 ± 0.4% vs. control 3.0...
Journal Articles
Journal: Diabetes
Diabetes 2004;53(12):3131–3141
Published: 01 December 2004
... demonstrated significantly higher levels of insulin production. Furthermore, streptozotocin-induced β-cell apoptosis (transferase-mediated dUTP nick-end labeling [TUNEL] assay) was significantly prevented in PID mice. Finally, PID mice exhibited a delayed onset of type 2 diabetes induced by a high-fat diet...
Journal Articles
Journal: Diabetes
Diabetes 2006;55(8):2245–2255
Published: 01 August 2006
... metalloproteinase NF-κB, nuclear factor-κB PCNA, proliferating cell nuclear antigen RAGE, receptor for AGE STZ, streptozotocin TSP, thrombospondin TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling VEGF, vascular endothelial growth factor The morbidity and mortality of diabetes...
Journal Articles
Journal: Diabetes
Diabetes 2007;56(1):80–87
Published: 01 January 2007
... (25 mmol/l glucose and 15% FCS) and deprived (5 mmol/l glucose and 1% FCS) conditions. Overexpression of gelsolin resulted in a decrease in the percentage of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)+ and active caspase-3+ cells. Conversely...
Journal Articles
Journal: Diabetes
Diabetes 2002;51(6):1938–1948
Published: 01 June 2002
.... In the hearts of diabetic mice, apoptotic cell death occurred as detected by terminal deoxynucleotidyl transferasemediated dUTP nick-end labeling (TUNEL) assay. Correspondingly, caspase-3 activation as determined by enzymatic assay and mitochondrial cytochrome c release detected by Western blotting...
Journal Articles
Journal: Diabetes
Diabetes 2009;58(2):329–336
Published: 01 February 2009
... β-cell number were determined by combined terminal deoxynucleotidyl transferasemediated dUTP nick-end labeling (TUNEL) staining and insulin immunohistochemistry. The in vitro effect of imatinib was studied using HepG2 cells. RESULTS— Imatinib induced remission of diabetes in db/db mice...
Journal Articles
Journal: Diabetes
Diabetes 2011;60(3):899–908
Published: 21 February 2011
..., immunohistochemistry, and transferase-mediated dUTP nick-end labeling (TUNEL) assay were performed. RESULTS In rats treated with MG and MG + l-buthionine sulfoximine (BSO), MG levels were significantly elevated in plasma, pancreas, adipose tissue, and skeletal muscle; fasting plasma glucose was elevated...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2010;59(7):1825–1835
Published: 27 April 2010
..., and CaMKII activity were all greatly increased in the retinas of diabetic mice compared with controls, 2 months after induction of diabetes. Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)-positive signals co-localized with CaMKII- and phospho-CaMKII immunoreactive RGCs...
Journal Articles
Journal: Diabetes
Diabetes 2010;59(2):440–447
Published: 29 October 2009
... a solution of vehicle (ethanol/water) mixed with fatty acid–free BSA at the same concentration as the palmitate solution. Staurosporine and thapsigargin were from Invitrogen and were dissolved in DMSO. For transferase-mediated dUTP nick-end labeling (TUNEL), ∼100 isolated mouse islets were mixed...
Includes: Supplementary data
Images
No evidence for transgene-driven, preautoimmune pSC destruction. <em>A</em>...
Published: 03 June 2010
FIG. 7. No evidence for transgene-driven, preautoimmune pSC destruction. A and B: hemotoxilin-eosin/terminal deoxynucleotidyl transferase–mediated dUTP nick-end labeling stain in 3-week-old wild-type (A) or GFAP-tg (B) mice without evidence of early cell death. C: Positive TUNEL stain of thymus. D: Pancreatic lymph node cells (2 × 105) from 3-week-old GFAP-tg or wild-type NOD mice failed to respond to GFAP stimulation (n = 3/group; P > 0.1). White bar, ovalbumin; green bar, GFAP; black bar, Concannavalin A. E: Insulin auto-antigen presentation of CD11c+ DCs purified from 3-week-old wild-type or GFAP-tg spleens. Cocultures of 2 × 104 DCs/well and 2 × 105 purified T-cells from 10-week-old wild-type NOD females were stimulated with insulin (n = 3/group; P > 0.1). White bar, ovalbumin; blue bar, insulin; black bar, Concannavalin A. wt, wild type. (A high-quality digital representation of this figure is available in the online issue.) FIG. 7. No evidence for transgene-driven, preautoimmune pSC destruction. A and B: hemotoxilin-eosin/terminal deoxynucleotidyl transferase–mediated dUTP nick-end labeling stain in 3-week-old wild-type (A) or GFAP-tg (B) mice without evidence of early cell death. C: Positive TUNEL stain of thymus. D: Pancreatic lymph node cells (2 × 105) from 3-week-old GFAP-tg or wild-type NOD mice failed to respond to GFAP stimulation (n = 3/group; P > 0.1). White bar, ovalbumin; green bar, GFAP; black bar, Concannavalin A. E: Insulin auto-antigen presentation of CD11c+ DCs purified from 3-week-old wild-type or GFAP-tg spleens. Cocultures of 2 × 104 DCs/well and 2 × 105 purified T-cells from 10-week-old wild-type NOD females were stimulated with insulin (n = 3/group; P > 0.1). White bar, ovalbumin; blue bar, insulin; black bar, Concannavalin A. wt, wild type. (A high-quality digital representation of this figure is available in the online issue.) More
Images
High glucose–<span class="search-highlight">mediated</span> mesangial cell growth inhibition and hypertrophy can ...
Published: 17 October 2011
FIG. 6. High glucose–mediated mesangial cell growth inhibition and hypertrophy can be reversed by activation of Nrf2. A: Cell growth of HRMCs incubated in NG, HG, HG+tBHQ, HG+SF, or HG+CA DMEM media was monitored in real-time for 96 h (upper panel = average; lower panel = average with error). B: Cell death and proliferation were assessed by transferase-mediated dUTP nick-end labeling (TUNEL) assay (positive control is treated with cisplatin at 18 μmol/L for 24 h) or Ki67 immunolabeling. HG media induced cell growth inhibition, which was alleviated by coculture with an Nrf2 activator. C: Cell size of HRMCs incubated in NG, HG, HG+tBHQ, HG+SF, or HG+CA DMEM media for 96 h was measured by forward light scatter/flow cytometry. Incubation in HG media induced cellular hypertrophy that was reduced with an Nrf2 activator. D: Cell area is reported from GFP-transfected HRMCs incubated in NG, HG, HG+tBHQ, HG+SF, or HG+CA DMEM media for 48 h. Total cell area increased with HG conditions but was significantly reduced in the presence of an Nrf2 activator. Data in D are expressed as mean ± SD (n = 100). *P < 0.05 compared with NG group. #P < 0.05 Nrf2 activators compared with HG alone. FSC-H, forward scatter. SSC-H, side scatter. (A high-quality color representation of this figure is available in the online issue.) FIG. 6. High glucose–mediated mesangial cell growth inhibition and hypertrophy can be reversed by activation of Nrf2. A: Cell growth of HRMCs incubated in NG, HG, HG+tBHQ, HG+SF, or HG+CA DMEM media was monitored in real-time for 96 h (upper panel = average; lower panel = average with error). B: Cell death and proliferation were assessed by transferase-mediated dUTP nick-end labeling (TUNEL) assay (positive control is treated with cisplatin at 18 μmol/L for 24 h) or Ki67 immunolabeling. HG media induced cell growth inhibition, which was alleviated by coculture with an Nrf2 activator. C: Cell size of HRMCs incubated in NG, HG, HG+tBHQ, HG+SF, or HG+CA DMEM media for 96 h was measured by forward light scatter/flow cytometry. Incubation in HG media induced cellular hypertrophy that was reduced with an Nrf2 activator. D: Cell area is reported from GFP-transfected HRMCs incubated in NG, HG, HG+tBHQ, HG+SF, or HG+CA DMEM media for 48 h. Total cell area increased with HG conditions but was significantly reduced in the presence of an Nrf2 activator. Data in D are expressed as mean ± SD (n = 100). *P < 0.05 compared with NG group. #P < 0.05 Nrf2 activators compared with HG alone. FSC-H, forward scatter. SSC-H, side scatter. (A high-quality color representation of this figure is available in the online issue.) More
Journal Articles
Journal: Diabetes
Diabetes 2004;53(12):3233–3238
Published: 01 December 2004
... the microvessels ( 3 , 8 ). ELISA, enzyme-linked immunosorbent assay NF-κB, nuclear transcriptional factor-κB 8-OHdG, 8-hydroxy-2′deoxyguanosine TUNEL, transferase-mediated dUTP nick-end labeling Diabetic retinopathy is the leading cause of acquired blindness among young adults, and studies have shown...
Journal Articles
Journal: Diabetes
Diabetes 2008;57(4):938–944
Published: 01 April 2008
... LacZ INS-TXNIP, INS-1 cells with constitutive TXNIP overexpression HBSS, Hanks' balanced salt solution TUNEL, transferase-mediated dUTP nick-end labeling TXNIP, thioredoxin-interacting protein Type 2 diabetes is a growing public health issue characterized by peripheral insulin resistance...
Journal Articles
Journal: Diabetes
Diabetes 2004;53(4):1096–1103
Published: 01 April 2004
..., streptozotocin TUNEL, transferase-mediated dUTP nick-end labeling VEGF, vascular endothelial growth factor Accelerated atherosclerosis makes peripheral ischemia dramatically common and severe in diabetic patients ( 1 , 2 ). In addition, clinical outcome after vascular occlusion is uniquely poor because...
Images
HDLs protect β-cells against apoptosis induced by ER stress. <em>A</em>...
Published: 13 April 2012
FIG. 1. HDLs protect β-cells against apoptosis induced by ER stress. A: Human islets from cadaveric donors were dissociated using trypsin and plated. The next day, islets were treated (TG) or not (control [C]) with 10 μmol/L TG in the presence (HDL) or in the absence (vehicle [VEH]) of HDLs for 24 h. Cells were then fixed, and apoptosis was assessed by scoring pycnotic nucleus. B: Cultured rat islets were incubated 24 h in serum-free RPMI 1640 medium containing 5 g/L BSA and 10 mmol/L glucose with the indicated combinations of 1 μmol/L TG and 1 mmol/L HDLs. Cell death was determined by transferase-mediated dUTP nick-end labeling (TUNEL) in insulin-expressing cells on histological sections of the islets. Results are expressed as the percentage of apoptotic cells among insulin-positive cells in a given islet section. A minimum of 2,000 cells from at least 20 islets have been scored from two independent experiments. C: MIN6 cells were treated with 0.5 μmol/L TG in the presence or in the absence of 1 mmol/L HDLs for 24 h. Cells were then fixed, and apoptosis was determined. Alternatively, the cells were lysed and the extent of caspase-3 activation was assessed by Western blotting using an antibody recognizing the cleaved active form of the protease. An actin-specific antibody was also used on the same blot to assess the evenness of loading. D: MIN6 cells were left untreated (control [Ctrl]) or treated for 48 h with 0.3% BSA (BSA) or 0.3% BSA/0.4 mmol/L palmitate (P) in the presence or in the absence of 1 mmol/L HDLs. Apoptosis was then scored as in A. E and F: Cultured rat islets were incubated for a week in serum-free RPMI 1640 medium containing 5 g/L BSA and 10 or 30 mmol/L glucose (labeled G10 and G30 in the figure) in the presence (HDL) or in the absence (vehicle [VEH]) of 1 mmol/L HDLs. Apoptosis was then assessed as in B. E: representative examples of TUNEL staining (green staining) in insulin-positive cells (red staining; nuclei are stained in blue with DAPI). The corresponding quantitation is shown in F. *Significant differences. (A high-quality digital representation of this figure is available in the online issue.) FIG. 1. HDLs protect β-cells against apoptosis induced by ER stress. A: Human islets from cadaveric donors were dissociated using trypsin and plated. The next day, islets were treated (TG) or not (control [C]) with 10 μmol/L TG in the presence (HDL) or in the absence (vehicle [VEH]) of HDLs for 24 h. Cells were then fixed, and apoptosis was assessed by scoring pycnotic nucleus. B: Cultured rat islets were incubated 24 h in serum-free RPMI 1640 medium containing 5 g/L BSA and 10 mmol/L glucose with the indicated combinations of 1 μmol/L TG and 1 mmol/L HDLs. Cell death was determined by transferase-mediated dUTP nick-end labeling (TUNEL) in insulin-expressing cells on histological sections of the islets. Results are expressed as the percentage of apoptotic cells among insulin-positive cells in a given islet section. A minimum of 2,000 cells from at least 20 islets have been scored from two independent experiments. C: MIN6 cells were treated with 0.5 μmol/L TG in the presence or in the absence of 1 mmol/L HDLs for 24 h. Cells were then fixed, and apoptosis was determined. Alternatively, the cells were lysed and the extent of caspase-3 activation was assessed by Western blotting using an antibody recognizing the cleaved active form of the protease. An actin-specific antibody was also used on the same blot to assess the evenness of loading. D: MIN6 cells were left untreated (control [Ctrl]) or treated for 48 h with 0.3% BSA (BSA) or 0.3% BSA/0.4 mmol/L palmitate (P) in the presence or in the absence of 1 mmol/L HDLs. Apoptosis was then scored as in A. E and F: Cultured rat islets were incubated for a week in serum-free RPMI 1640 medium containing 5 g/L BSA and 10 or 30 mmol/L glucose (labeled G10 and G30 in the figure) in the presence (HDL) or in the absence (vehicle [VEH]) of 1 mmol/L HDLs. Apoptosis was then assessed as in B. E: representative examples of TUNEL staining (green staining) in insulin-positive cells (red staining; nuclei are stained in blue with DAPI). The corresponding quantitation is shown in F. *Significant differences. (A high-quality digital representation of this figure is available in the online issue.) More
Journal Articles
Journal: Diabetes
Diabetes 2003;52(7):1701–1708
Published: 01 July 2003
... m-IAPP, mouse IAPP TLVM, time-lapse video microscopy TUNEL, transferase-mediated dUTP nick-end labeling Type 2 diabetes is characterized by partial β-cell loss ( 1 – 3 ) and islet amyloid derived from the 37-amino acid peptide islet amyloid polypeptide (IAPP) ( 4 – 8 ). IAPP is coexpressed...
Journal Articles
Journal: Diabetes
Diabetes 2005;54(2):540–545
Published: 01 February 2005
... (data not shown). Therefore, we used all cells after a 48-h period of serum withdrawal. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and caspase-3 activity assays were used as quantitative indexes of apoptosis. Apoptosis was detected using fluorescein...
Journal Articles
Journal: Diabetes
Diabetes 2003;52(11):2731–2739
Published: 01 November 2003
... of rapamycin PI, propidium iodide TUNEL, transferase-mediated dUTP nick-end labeling DIABETES 2003 7 8 2003 28 6 2002 A two-color fluorescence cell viability assay was used based on the ability of calcein AM to be retained within live cells, inducing an intense uniform green...