Evaluation of safety and efficacy of adipose-derived stem cells in rat myocardial infarction model using hexadecyl-4-[¹²4I]iodobenzoate for cell tracking.

This study was aimed to evaluate the effect of (124)I-labeling with hexadecyl-4-iodobenzoate (HIB) on gene expression related to cell cycle, DNA repair, transcription, proliferation and differentiation of adipose-derived stem cells (ADSCs). [(124)I]HIB showed high labeling efficiency with ADSCs (51.3±1.3%, 0.3-2.0 Bq/cell) and there is no morphological change of ADSCs. In the microarray analysis of gene expression pattern, differences were not observed between non-labeled and [(124)I]HIB-labeled ADSCs. We demonstrated that (124)I-labeling with HIB did not affect the biological properties of ADSCs.

Abdominal adipocyte populations in women with visceral obesity.

BACKGROUND: Visceral obesity is independently related to numerous cardiometabolic alterations, with adipose tissue dysfunction as a central feature. OBJECTIVE: To examine whether omental (OM) and subcutaneous (SC) adipocyte size populations in women relate to visceral obesity, cardiometabolic risk factors and adipocyte lipolysis independent of total adiposity. DESIGN AND METHODS: OM and SC fat samples were obtained during gynecological surgery in 60 women (mean age, 46.1±5.9 years; mean BMI, 27.1±4.5 kg/m² (range, 20.3-41.  kg/m²)). Fresh samples were treated with osmium tetroxide and were analyzed with a Multisizer Coulter. Cell size distributions were computed for each sample with exponential and Gaussian function fits. RESULTS: Computed tomography-measured visceral fat accumulation was the best predictor of larger cell populations as well as the percentage of small cells in both OM and SC fat (P<0.0001 for all). Accordingly, women with visceral obesity had larger cells in the main population and higher proportion of small adipocytes independent of total adiposity (P≤0.05). Using linear regression analysis, we found that women characterized by larger-than-predicted adipocytes in either OM or SC adipose tissue presented higher visceral adipose tissue area, increased percentage of small cells and homeostasis model assessment insulin resistance index as well as higher OM adipocyte isoproterenol-, forskolin- and dbcAMP-stimulated lipolysis compared to women with smaller-than-predicted adipocytes, independent of total adiposity (P≤0.05). CONCLUSION: Excess visceral adipose tissue accumulation is a strong marker of both adipocyte hypertrophy and increased number of small cells in either fat compartment, which relates to higher insulin resistance index and lipolytic response, independent of total adiposity.

Adenovirus-mediated expression of human sodium-iodide symporter gene permits in vivo tracking of adipose tissue-derived stem cells in a canine myocardial infarction model.

INTRODUCTION: In vivo tracking of the transplanted stem cells is important in pre-clinical research of stem cell therapy for myocardial infarction. We examined the feasibility of adenovirus-mediated sodium iodide symporter (NIS) gene to cell tracking imaging of transplanted stem cells in a canine infarcted myocardium by clinical single photon emission computed tomography (SPECT). METHODS: Beagle dogs were injected intramyocardially with NIS-expressing adenovirus-transfected canine stem cells (Ad-hNIS-canine ADSCs) a week after myocardial infarction (MI) development. (99m)Tc-methoxyisobutylisonitrile ((99m)Tc-MIBI) and (99m)Tc-pertechnetate ((99m)TcO4(-)) SPECT imaging were performed for assessment of infarcted myocardium and viable stem cell tracking. Transthoracic echocardiography was performed to monitor any functional cardiac changes. RESULTS: Left ventricular ejection fraction (LVEF) was decreased after LAD ligation. There was no significant difference in EF between the groups with the stem cell or saline injection. (125)I uptake was higher in Ad-hNIS-canine ADSCs than in non-transfected ADSCs. Cell proliferation and differentiation were not affected by hNIS-carrying adenovirus transfection. (99m)Tc-MIBI myocardial SPECT imaging showed decreased radiotracer uptake in the infarcted apex and mid-anterolateral regions. Ad-hNIS-canine ADSCs were identified as a region of focally increased (99m)TcO4(-) uptake at the lateral wall and around the apex of the left ventricle, peaked at 2 days and was observed until day 9. CONCLUSIONS: Combination of adenovirus-mediated NIS gene transfection and clinical nuclear imaging modalities enables to trace the fate of transplanted stem cells in infarcted myocardium for translational in vivo cell tracking study for prolonged duration.

Low-intensity pulsed ultrasound induced enhanced adipogenesis of adipose-derived stem cells.

OBJECTIVE: The aim of this study was to investigate effects of low-intensity pulsed ultrasound (LIPUS) on differentiation of adipose-derived stem cells (ASCs), in vitro. MATERIALS AND METHODS: Murine ASCs were treated with LIPUS for either three or five days, immediately after adipogenic induction, or delayed for 2 days. Expression of adipogenic genes PPAR-γ1, and APN, was examined by real-time PCR. Immunofluorescence (IF) staining was performed to test for PPAR-γ at the protein level. RESULTS: Our data revealed that specific patterns of LIPUS up-regulated levels of both PPAR-γ1 and APN mRNA, and PPAR-γ protein. CONCLUSIONS: In culture medium containing adipogenic reagents, LIPUS enhanced ASC adipogenesis.

Evaluation of a novel high-intensity focused ultrasound device for ablating subcutaneous adipose tissue for noninvasive body contouring: safety studies in human volunteers.

BACKGROUND: High-intensity focused ultrasound (HIFU) is an energy-based medical technology with many clinical applications. A device under clinical investigation in the United States (LipoSonix; Medicis Technologies Corporation, Bothell, Washington) uses HIFU to reduce localized deposits of abdominal adipose tissue. OBJECTIVES: The authors describe the results from their clinical trial investigating the safety of this HIFU device in human patients. METHODS: Over the course of three studies evaluating the safety of the HIFU device for ablating human subcutaneous adipose tissue (SAT), 152 healthy patients were treated with total HIFU energy doses of 47 to 331 J/cm(2)), including patients who presented for elective abdominoplasty and underwent treatment to areas identified for subsequent excision. The safety of each treatment regimen was confirmed before the energy levels were raised. Abdominoplasty was performed up to 14 weeks following the HIFU procedure, and a pathologist performed histopathological analyses of excised tissues. Safety evaluations included an assessment of clinical chemistry and hematology profiles, physical examinations, and adverse events. RESULTS: Posttreatment ultrasound confirmed that the HIFU effects were limited to targeted SAT layers. Histopathology revealed well-demarcated disruption of adipocytes within the targeted SAT. Phagocytosis of released lipids and cellular debris occurred after 14 to 28 days. Phagocytized lipids underwent normal hepatic metabolism. Healing progressed normally and was 95% complete after eight to 14 weeks. Adverse events consisted primarily of temporary treatment discomfort, edema, erythema, dysesthesia, and ecchymosis. There were no changes in clinical laboratory parameters, and no serious device-related adverse events occurred. Optimal clinical outcomes were achieved with lower energy levels, which provided beneficial effects with the least amount of discomfort. CONCLUSIONS: HIFU appears to provide a safe means for removing and remodeling unwanted deposits of abdominal SAT.

Multiplex RARE: a simultaneous multislice spin-echo sequence that fulfils CPMG conditions.

This work presents a new imaging sequence in which multiple slices are simultaneously excited and refocused in a spin-echo train. The multiple spin-echo trains are interleaved in such a manner that (i) the Carr-Purcell-Meiboom-Gill conditions are fulfilled at all times, and (ii) the signals from slices can be separated, preventing aliasing. This paper also demonstrates how the sequence may be used in a novel fat-water Dixon method that enables fast volume coverage. The technique is demonstrated in phantoms and in vivo.

Intracoronary administration of autologous adipose tissue-derived stem cells improves left ventricular function, perfusion, and remodelling after acute myocardial infarction.

AIMS: This study was designed to assess whether intracoronary application of adipose tissue-derived stem cells (ADSCs) compared with bone marrow-derived stem cells (BMSCs) and control could improve cardiac function after 30 days in a porcine acute myocardial infarction/reperfusion model. METHODS AND RESULTS: An acute transmural porcine myocardial infarction was induced by inflating an angioplasty balloon for 180 min in the mid-left anterior descending artery. Two million cultured autologous stem cells were intracoronary injected through the central lumen of the inflated balloon catheter. Analysis of scintigraphic data obtained after 28 +/- 3 days showed that both absolute and relative perfusion defect decreased significantly after intracoronary administration of ADSCs or BMSCs (relative 30 or 31%, respectively), compared with carrier administration alone (12%, P = 0.048). Left ventricular ejection fraction after 4 weeks increased significantly more after ADSC and BMSC administration than after carrier administration: 11.39 +/- 4.62 and 9.59 +/- 7.95%, respectively vs. 1.95 +/- 4.7%, P = 0.02). The relative thickness of the ventricular wall in the infarction area after cell administration was significantly greater than that after carrier administration. The vascular density of the border zone also improved. The grafted cells co-localized with von Willebrand factor and alpha-smooth muscle actin and incorporated into newly formed vessels. CONCLUSION: This is the first study to show that not only bone marrow-derived cells but also ADSCs engrafted in the infarct region 4 weeks after intracoronary cell transplantation and improved cardiac function and perfusion via angiogenesis.

[Function of PAT family proteins in the lipid metabolism].

Triglycerides are stored in intracellular lipid droplets in mammalian cells. Current evidences suggest that the intracellular lipid droplets are active participants in a variety of metabolic processes, and thus are considered as functional organelles in cells. The lipid droplet is composed of a triglyceride core surrounded by a phospholipid monolayer in which several proteins are embedded. Four of these proteins belong to a family of structurally related PAT protein, which includes perilipin, ADRP, TIP47 and S3-12. In this review, we summarize the functions of these proteins in the modulation of lipolysis and lipid droplets formation.

Biological studies of radiolabeled glucose analogues iodinated in positions 3, 4 or 6.

The aim of this study was to assess the biological behavior of new radiolabeled glucose analogues proposed as tracers of glucose uptake in vivo and iodinated in position 3, 4, or 6. Biological results obtained in vitro on adipocytes and erythrocytes and in vivo in mice were compared to those obtained with the gold-standard tracer of glucose uptake, 2-deoxy-D-glucose. None of these molecules had the same biological behavior than 2-deoxy-D-glucose. Therefore, these compounds cannot be considered as tracers of glucose uptake.

Ultrasonographic assessment of coronary flow reserve and abdominal fat in obesity.

Recent technological advances in transthoracic Doppler echocardiography (TTDE) have provided noninvasive measurement of coronary flow velocity reserve (CFVR). We aimed to quantitate a correlation between endothelial dysfunction and fat distribution. In 36 patients with obesity, 16 with noninsulin-dependent diabetes mellitus (DM) and 12 healthy volunteers, coronary flow velocity was measured at the distal site of the left anterior descending branch. CFVR was defined as the ratio of hyperemic (IV infusion of 0.15 mg/kg/min adenosine) to basal peak diastolic flow velocity. Abdominal wall fat index (AWFI) was estimated by ultrasonography. Insulin resistance was quantified by the euglycemic hyperinsulinemic clump method. AWFI was significantly related to CFVR (r = -0.46, p = 0.011) and insulin resistance (r = -0.71, p < 0.0001). CFVR could be noninvasively evaluated using TTDE. Coronary endothelial dysfunction indicated as CFVR, body fat distribution and insulin resistance was quantitatively correlated in obesity.

Glucocorticoid-induced insulin resistance associates with activation of protein kinase C isoforms.

We studied glucocorticoid-induced insulin resistance and possible role of protein kinase C (PKC). Pretreatment with dexamethasone, prednisolone and corticosterone for 60 min decreased insulin-induced [3H] 2-deoxyglucose (DOG) uptake in isolated rat adipocytes. Preincubation with Go6976, LY379196 or myristoylated PKC pseudosubstrate, conventional PKC inhibitor, but not cycloheximide or RU38486, recovered dexamethasone-induced insulin resistance. Dexamethasone activated immunoprecipitates with anti-PKC alpha, beta, and zeta antibodies. PKC zeta activity in adipocytes increased to 163%, and 264% from basal level (100%) with dexamethasone and insulin treatment, respectively. Dexamethasone provoked redistribution of both PKC beta and zeta from the cytosol to the membrane. These results indicate that dexamethasone activates both conventional and atypical PKC. However, conventional PKC is more important in glucocorticoid-induced insulin resistance.