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A Gagnon

Publications and source records attributed to A Gagnon.

At least 37 records · Page 2Linked to original sources
PubMed1998

The effect of glucose concentration on insulin-induced 3T3-L1 adipose cell differentiation.

We examined the effect of glucose concentration on insulin-induced 3T3-L1 adipose cell differentiation. Oil Red O staining of neutral lipid, cellular triglyceride mass, and glycerol phosphate dehydrogenase (GPDH) activity, were greater in 3T3-L1 cells cultured at 5 mM vs. 25 mM glucose. GPDH activity was 2- to 4-fold higher at 5 mM vs. 25 mM glucose over a range of insulin concentrations (0.1 to 100 nM). Insulin-stimulated tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) was 1.7-fold greater, and insulin-stimulated phosphoinositide 3-kinase association with IRS-1 was 2.3-fold higher, at 5 mM vs. 25 mM glucose. These effects of glucose were not caused by alterations in IRS-1 mass or cell-surface insulin binding. In preadipose cells at 5 mM glucose, expression of the leukocyte antigen-related (LAR) protein tyrosine phosphatase (negative regulator of insulin signaling) was 63% of the level at 25 mM glucose. Our data demonstrate that glucose concentration affects insulin-induced 3T3-L1 adipose cell differentiation as well as differentiation-directed insulin signaling pathways. Alterations in LAR expression potentially may be involved in modulating these responses.

3T3 Cells↗
PubMed1998

Endoscopic laser coagulation in the management of severe twin-to-twin transfusion syndrome.

OBJECTIVE: To assess the clinical effectiveness of endoscopic laser coagulation of placental vessels in the treatment of severe transfusion syndrome. DESIGN: Prospective study. SETTING: Three referral centres for the management of twin-to-twin transfusion syndrome. POPULATION: One hundred and thirty-two pregnancies complicated by severe twin-to-twin transfusion syndrome, reflected by polyhydramnios and enlarged bladder of one twin and oligoanhydramnios and collapsed bladder of the other twin, presenting before 28 weeks of gestation. METHODS: Prospective collection of data on pre-procedure assessment, the procedure and the follow up were collected prospectively. Laser coagulation of placental vessels crossing the intertwin membrane on the chorionic surface under sono-endoscopic guidance, followed by amniodrainage. MAIN OUTCOME MEASURES: Maternal and pregnancy complications, perinatal death and morbidity were assessed over the last five years with follow up of survivors. RESULTS: Endoscopic laser was carried out at a median gestation of 21 weeks. The total number of surviving infants was 144 (55%) and there was at least one survivor in 97 cases (73%). At a minimum age of one year neurological handicap was suspected in six survivors (4.2%). CONCLUSIONS: The results of this multicentre study are similar to those in our original report on the first 45 cases. In comparison with serial amniodrainage, the survival rate may be similar, but the handicap rate in survivors appears much lower. This study stresses the need for a prospective study comparing these two techniques.

Endoscopy↗
PubMed1998

Expression and regulation of neuronal apoptosis inhibitory protein during adipocyte differentiation.

We have used the 3T3-L1 and 3T3-F442A preadipocyte cell lines to examine the expression and regulation of neuronal apoptosis inhibitory protein (NAIP) during adipocyte differentiation. When 3T3-L1 preadipocytes differentiated into adipocytes, they developed resistance to apoptosis induced by growth factor deprivation, as assessed by terminal deoxynucleotide transferase (TdT)-mediated dUTP nick end labeling. Protein expression of NAIP was markedly elevated in 3T3-L1 and 3T3-F442A adipocytes compared with that in their fibroblast-like precursors. NAIP was also present in rat white adipocytes. In 3T3-L1 cells, the increase in NAIP occurred by day 4 of the 8-day differentiation protocol, which includes exposure of confluent preadipocytes to insulin, dexamethasone, and isobutylmethylxanthine. Incubation of confluent 3T3-L1 preadipocytes with any of these components alone had no effect on NAIP expression. When 3T3-C2 cells, a control cell line that does not differentiate, were subjected to the differentiation protocol, the low NAIP levels remained unaltered. Addition of rapamycin, a p70 S6 kinase inhibitor that blocks adipocyte differentiation, to the 3T3-L1 differentiation medium prevented the rise in NAIP expression. These data demonstrate for the first time that NAIP is expressed in adipocyte cell lines and primary adipocytes. The differentiation-dependent augmentation of NAIP protein levels in 3T3-L1 adipocytes is closely correlated with the development of resistance to apoptosis induced by growth factor deprivation, suggesting a potential role for NAIP in these cells.

3T3 Cells↗
PubMed1998

Fasting and nonfasting iodine-123-idophenylpentadecanoic acid myocardial SPECT imaging in coronary artery disease.

UNLABELLED: Iodine-123-labeled idophenylpentadecanoic acid (IPPA) metabolic imaging has been shown to be clinically useful for the identification of myocardial viability in patients with coronary artery disease and left ventricular dysfunction. Imaging is usually performed under fasting conditions since nonfasting conditions may affect myocardial uptake of 123I-IPPA. The purpose of this study was to examine the impact of dietary condition on 123I-IPPA metabolic imaging. METHODS: Forty patients with stable coronary artery disease underwent, in randomized order and on separate days, 123I-IPPA SPECT myocardial imaging under fasting and nonfasting conditions. Patients were injected with 123I-IPPA (4-5 mCi) at rest with imaging performed at 4 (initial) and 30 (delay) min. For each image (initial and delay images), 10 segments were analyzed by three experienced observers without knowledge of patient identity or dietary condition using a 5-point grading system (O = no uptake to 4 = normal uptake). A summed global score was obtained for each image by adding the scores for all 10 segments. Image quality was assessed using a 3-point grading system. RESULTS: Visual agreement for normal and abnormal segments between fasting and nonfasting conditions was 82% (kappa = 0.63). There were no significant differences in the summed global scores for both conditions. Image quality was equivalent for both conditions in 65% of cases and superior under the nonfasting condition in 25% of cases. CONCLUSION: Image quality as well as the presence, location and severity of defects are similar under fasting and nonfasting conditions with 123I-IPPA. Therefore, fasting is not necessary before 123I-IPPA SPECT imaging for the assessment of myocardial viability.

Adult↗
PubMed1997

Modulation and phosphorylation of calbindin-D28K correlates with protein kinase C activation1.

Calbindin-D28K is a vitamin D3 dependent calcium-binding protein expressed in renal distal tubules. We previously reported that 24 h treatment with 1 alpha, 25-dihydroxyvitamin D3 (1,25-D3), the active form of vitamin D3, induces calbindin-D28K and activates protein kinase C (PKC) in MDBK (Madin-Darby bovine kidney) cells. In contrast, 24 h treatment with the phorbol ester 12-O-tetradecanoylphorbol-3-acetate (TPA) downregulates calbindin-D28K and PKC activity. In the present studies, we demonstrate that TPA rapidly enhances calbindin-D28K expression in MDBK cells in the absence of 1,25-D3. The enhancement of calbindin-D28K expression is preceded by activation and translocation of PKC alpha. Further, we show that PKC directly phosphorylates calbindin-D28K in a calcium- and phospholipid-dependent manner in vitro. In MDBK cells, the calbindin-D28K antibody immunoprecipitates a 28 kDa protein for which phosphorylation is enhanced after treatment for 1 h with TPA or 24 h with 1,25-D3. Consistent with amino acid sequence analysis of calbindin-D28K indicating two threonine residues that fit the consensus for PKC phosphorylation, TPA-treated MDBK cells exhibit enhanced expression of a phosphothreonine-containing protein that co-migrates with calbindin-D28K. These studies offer the first report that calbindin-D28K is a phosphoprotein and implicate the PKC signal transduction pathway in its regulation.

Animals↗
PubMed1997

[Surgical treatment of twin-to-twin transfusion syndrome: coagulation of anastomoses with a Nd:YAG laser, under endosonographic control. Forty four cases].

Twin to twin transfusion syndrome occurs in 15% of monozygotic twin pregnancies and is characterized by a hemodynamic imbalance between the two fetuses. Survival is about 10% with expectant management. Our aim is to coagulate the placental vascular anastomoses joining the two fetal circulations on the placenta in order to prevent the consequences on the survivor when one dies in utero. Between November 1993 and April 1996, 44 patients were referred with severe twin to twin transfusion syndrome at less than 28 weeks' gestation. Nd:YAG laser coagulation of the placental vessels crossing the interamniotic membrane was performed under sono-endoscopic control. Forty-one patients have completed their pregnancy and 3 are continuing uneventfully in the third trimester. Fifteen patients (36.5%) delivered 2 healthy infants who survived the neonatal period and are developing normally. Sixteen patients (41.5%) delivered one infant developing, normally after the death of the co-twin in the perinatal period. In ten cases (22%) both twins died in the perinatal period. Among the survivors, 3 (9%) present with neurological handicap at a median age of 9 months. Around 75% (31/41) pregnancies complicated by twin to twin transfusion and treated by fetoscopy delivered at least one healthy baby. These data confirm the feasibility of the technique and that morbidity in-the survivors is relatively low.

Endosonography↗
PubMed1996

Evidence of adipocyte differentiation in human orbital fibroblasts in primary culture.

Graves' ophthalmopathy (GO), in which orbital tissues are infiltrated with activated T lymphocytes and hyaluronan, can manifest an overabundance of adipose tissue in the human orbit. Little is known about adipogenesis in this anatomic region. We have investigated whether orbital fibroblasts in culture possess the capacity to undergo adipocytic differentiation. Orbital tissue from patients with or without GO was placed in primary culture, and proliferating fibroblasts were sub-passaged. Confluent fibroblasts were subjected to a differentiation protocol, involving a serum-free defined medium supplemented with insulin, triiodothyronine, carbaprostacyclin, thyrotropin, dexamethasone, and isobutylmethylxanthine. Control cells were maintained in serum-free medium supplemented only with insulin. After approximately 14 days, light microscopy revealed characteristic morphologic changes of adipocyte differentiation, including cell rounding and lipid droplet accumulation. Oil Red O staining could be demonstrated in those cells. Scanning electron microscopy revealed that the cells undergoing adipogenesis contain multiple, discrete lipid droplets. The overall percentage of fibroblasts undergoing differentiation was somewhat variable, but no more than 5-10%. The adipocytes arise in close proximity to each other, and often at the periphery of the culture surface. In contrast, dermal fibroblasts and perimysial fibroblasts from extraocular muscle fail to differentiate. We conclude that a subpopulation of orbital fibroblasts is capable of adipocyte differentiation. These in vitro observations may represent the in vivo process that contributes to excess orbital adipose tissue volume in GO.

Adipocytes↗
PubMed1994

1,25-Dihydroxyvitamin D3 translocates protein kinase C beta to nucleus and enhances plasma membrane association of protein kinase C alpha in renal epithelial cells.

1,25-Dihydroxycholecalciferol (1,25-(OH)2-D3) increases membrane-associated protein kinase C (PKC) activity and immunoreactivity in renal epithelial (Madin Darby bovine kidney, MDBK) cells (Simboli-Campbell, M., Franks, D. J., and Welsh, J. E. (1992) Cell Signalling 4, 99-109). We have now characterized the effects of 1,25-(OH)2-D3 on the subcellular localization of three individual isozymes by immunofluorescence and immunoblotting. Although the total amount of PKC alpha, PKC beta, and PKC zeta are unaffected by 1,25-(OH)2-D3, this steroid hormone induces subcellular redistribution of both PKC alpha and PKC beta. Treatment with 1,25-(OH)2-D3 (100 nM, 24 h) enhances plasma membrane association of PKC alpha and induces translocation of PKC beta to the nuclear membrane. The effects of 1,25-(OH)2-D3 appear to be limited to the calcium-dependent PKC isozymes, since 1,25-(OH)2-D3 has no effect on the calcium independent isozyme, PKC zeta. In contrast to rapid transient PKC translocation seen in response to agents which interact with membrane receptors to induce phospholipid hydrolysis, modulation of PKC alpha and PKC beta is observed after 24 h treatment with 1,25-(OH)2-D3. In MDBK cells, the phorbol ester 12-0-tetradecanoylphorbol-13-acetate (TPA) (100 nM, 24 h) down-regulates PKC alpha and, to a lesser extent, PKC zeta, without altering their subcellular distribution. TPA also induces translocation of PKC beta to the nuclear membrane. MDBK cells treated with 1,25-(OH)2-D3, but not TPA, exhibit enhanced phosphorylation of endogenous nuclear proteins. In addition to the distinct effects of 1,25-(OH)2-D3 and TPA on PKC isozyme patterns, 1,25-(OH)2-D3 up-regulates both the vitamin D receptor and calbindin D-28K, whereas TPA down-regulates the expression of both proteins. These data support the involvement of PKC in the mechanism of action of 1,25-(OH)2-D3 and specifically implicate PKC beta in 1,25-(OH)2-D3-mediated nuclear events.

Animals↗
PubMed1994

Neutropenic fever in patients after high-dose chemotherapy followed by autologous haematopoietic progenitor cell transplantation and human recombinant granulocyte-macrophage colony stimulating factor.

Neutropenic fever has been one of the most difficult complications in the recovery period following high-dose chemotherapy and autologous haematopoietic progenitor cell transplantation. The differentiation between human recombinant GM-CSF (sargramostim)-related fever and active infection can be difficult during this observation time. In 7 of 17 patients treated for metastatic breast cancer with HDCT and PBPC within 6 consecutive months, neutropenic fever without signs of infection was observed, which may be sargramostim-related fever. The typical presentation must fulfil the following criteria: cyclical elevation in body temperature that happens at the predicted time after sargramostim administration; absence of other signs or symptoms of infections; quick resolution of the fever after onset acetaminophen administration. Having met these criteria, none of these patients has been treated with intravenous antibiotics for active infections. At the time of haematological recovery (at a median time of 13 days from PBPC reinfusion to absolute neutrophil counts of > or = 0.5/nl) the febrile episode gradually resolved. No serious complications or other side-effects were observed. No toxic deaths occurred. Only if specific symptoms or signs of infection develop, would intravenous empiric antibiotic therapy be started.

Breast Neoplasms↗
PubMed1991

Dipyridamole-thallium imaging: minimum dose requirements.

A total of 372 consecutive dipyridamole-thallium studies were reviewed to determine if a relationship existed between the dose of dipyridamole administered and the likelihood that thallium images would show reversible defects. Men who received a dose of less than 31 mg were less likely to have thallium redistribution (P = 0.0001). Until the ideal study involving repeat testing with incremental dipyridamole doses on different days is carried out, the authors recommend that a minimal dose of 31 mg be administered for dipyridamole-thallium imaging.

Coronary Vessels↗
PubMed1990

Transient left ventricular cavitary dilation during dipyridamole-thallium imaging as an indicator of severe coronary artery disease.

Transient left ventricular (LV) cavitary dilation during dipyridamole-thallium imaging was reported in 45 of 510 (9%) consecutive patients referred for dipyridamole-thallium imaging. Clinical and hemodynamic effects observed during dipyridamole infusion were not predictive of transient cavitary dilation on the thallium images. Coronary angiography was performed in 32 of the 45 patients: 75% had either left main, 3-vessel or "high-risk" 2-vessel coronary artery disease. Although 25 of 45 patients (56%) with transient cavitary dilation were either asymptomatic or had only grade 1/4 effort angina, 16 of 25 patients (64%) not referred for coronary revascularization sustained a cardiac event during a mean follow-up of 12 months. Most events were cardiac deaths (75%) and 87% of events occurred within 4 months of the test. Noncardiac surgery was performed in 187 of the 510 patients. The postoperative cardiac event rate was 2% in the 101 patients with normal scans or fixed defects, 19% in 75 patients with reversible perfusion defects and 58% in 12 patients with reversible cavitary dilation (p less than 0.0001). Thus, transient LV dilation during dipyridamole-thallium imaging is a marker of severe underlying coronary artery disease, denotes a poor prognosis and predicts a high risk of postoperative cardiac complications in patients who undergo noncardiac surgery.

Coronary Angiography↗
PubMed1989

Usefulness of the severity and extent of reversible perfusion defects during thallium-dipyridamole imaging for cardiac risk assessment before noncardiac surgery.

Thallium-dipyridamole imaging is a very sensitive test for predicting cardiac events after noncardiac surgery, but it lacks specificity. To improve specificity, a semiquantitative scoring system was developed that combined dipyridamole-induced reversible left ventricular dilatation with scintigraphic indexes for severity and extent of reversible perfusion defects. Using this scoring system, patients were classified into low, intermediate and high risk subgroups. Thallium-dipyridamole imaging was performed in 66 patients before major general and vascular surgery. Thirty-nine patients classified as low risk (30 with normal scans and 9 with fixed defects) underwent surgery uneventfully. Surgery was cancelled in 6 patients with extensive thallium redistribution and coronary angiography was performed because of severe coronary artery disease in 5 and idiopathic dilated cardiomyopathy in 1. In the remaining 21 patients with thallium redistribution, a positive statistical correlation (p = 0.001) between scintigraphic indexes of severity and extent, and cardiac events was noted. Using cutoff values for the scintigraphic indexes, patients with reversible defects could be classified into intermediate and high risk subgroups. Only 1 of 11 patients at intermediate risk developed a complication, whereas 8 of 10 patients at high risk had a postoperative event (7 deaths and 1 myocardial infarction). Thus, using scintigraphic indexes for severity and extent, patients with reversible defects can be stratified into an intermediate risk subgroup that can safely undergo surgery and a high risk subgroup that requires coronary angiography.

Angiography↗
PubMed1989

Same day injections of Tc-99m methoxy isobutyl isonitrile (hexamibi) for myocardial tomographic imaging: comparison between rest-stress and stress-rest injection sequences.

It has been shown that both rest and stress 99mTc-hexamibi myocardial perfusion imaging can be performed on the same day using two different doses injected within few hours (the first one at rest followed by a second at stress). In order to evaluate and compare 2 sequences (rest-stress and stress-rest) of 99mTc-hexamibi injections performed the same day, 18 patients with either abnormal 201Tl myocardial scan or abnormal coronary angiography were studied with 2 99mTc-hexamibi injections protocols. The rest-stress study was performed as follows: 7 mCi 99mTc-hexamibi was injected at rest. Single photon emission computed tomography (SPECT) was performed 60 min later. Immediately after the rest study, patients were injected at peak stress with 25 mCi 99mTc-hexamibi. Tomographic imaging was repeated 1 h later. Patients were submitted to the stress-rest protocol within 3 days. Tomographic imaging was done 1 h after a 7 mCi injection at stress. This study was followed by an injection of 25 mCi 99mTc-hexamibi at rest, a tomographic study was performed 60 min later. Myocardial sections were reconstructed in horizontal long, vertical long, and short axes. Data analysis also included polar map representation. A total of 324 segments were interpreted blind by 3 observers, there was an agreement in 283/324 (87.3%) segments between the 2 protocols. However, 24 segments (7.4%) judged ischemic on rest-stress were called scars on stress-rest.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗