Publications

Bone marrow-derived mesenchymal stem cells (MSC) are a promising source for cell-based treatment of myocardial infarction (MI), but existing strategies are restricted by low cell survival and engraftment. We examined whether vascular endothelial growth factor (VEGF) improve MSC viability in infarcted hearts. We found long-term culture increased MSC-cellular stress: expressing more cell cycle inhibitors, p16(INK), p21 and p19(ARF). VEGF treatment reduced cellular stress, increased pro-survival factors, phosphorylated-Akt and Bcl-xL expression and cell proliferation. Co-injection of MSCs with VEGF to MI hearts increased cell engraftment and resulted in better improvement of cardiac function than that injected with MSCs or VEGF alone. In conclusion, VEGF protects MSCs from culture-induce cellular stress and improves their viability in ischemic myocardium, which results in improvements of their therapeutic effect for the treatment of MI.

BACKGROUND

Rodents with deficiency of or resistance to the proopiomelanocortin-derived peptide gamma-melanocyte stimulating hormone (gamma-MSH) develop marked salt-sensitive hypertension. We asked whether this hypertension was accompanied by abnormal glucose metabolism.

METHODS

gamma-MSH-deficient Pc2(-/-) mice, and resistant Mc3r(-/-) mice were studied acutely for measurement of blood pressure and glucose and insulin concentrations after > or =1 week of a high-sodium diet (HSD; 8% NaCl) compared to a normal-sodium diet (NSD; 0.4% NaCl). Mc3r(-/-) also underwent glucose tolerance test (GTT) and insulin tolerance test.

RESULTS

Both knockout strains were hypertensive and also exhibited fasting hyperglycemia and hyperinsulinemia on the HSD. Mc3r(-/-) mice on the HSD had impaired glucose tolerance and insulin-mediated glucose disposal compared to wild-type mice on either the HSD or the NSD, or to Mc3r(-/-) mice on the NSD.

CONCLUSIONS

These results indicate an interaction of interrupted gamma-MSH signaling with the HSD to cause hypertension on the one hand and abnormal glucose metabolism, with the characteristics of insulin resistance, on the other. Further study of the nature of this interaction should provide new insight into the mechanisms by which salt-sensitive hypertension and insulin resistance are linked.

INTRODUCTION

Different technologies have been developed for radiofrequency ablation (RFA), which include increasing electrode (tip) size and cooling the tip through irrigation either internally (closed-loop) with D5W or externally (open-loop) with saline. Although these catheters are widely used clinically, the propensity for adverse events and the lesion profiles of each of these catheter technologies have not been directly compared under a wide range of controlled conditions.

METHODS AND RESULTS

Freshly excised canine thigh muscle was placed in a chamber filled with circulating, heparinized blood heated to 37 degrees C. Five different catheters were tested: 4 mm tip, 10 mm tip single thermistor, 10 mm tip multitemperature sensor, 4 mm closed-loop irrigated cooled-tip, and 4 mm open-loop irrigated cooled tip at several different contact and power settings. The catheter and tissue interface was continuously monitored with intracardiac echocardiography (echo) (Acuson). During the RFA, any bubbling generated from the tip and/or popping seen on echo was noted, and after each RFA, the catheter and lesion were examined for the presence of thrombus. For all of the catheters, complications correlated to the electrode tip temperature and power setting. All of the catheters experienced complications at any lesion size except for the open-irrigated catheter, which only had complications at the largest lesions. Overall, the cooled tip catheters experienced an at least sixfold greater odds of popping, bubbling, and impedance rises than the 4 mm, but the majority occurred at power levels greater than 20 W. The open-irrigated catheters created eccentric lesions that extended away from the tissue-catheter interface, in the direction of blood flow. In addition, it produced saline filled blisters at the lesion site in 16.7% of the burns. The 10 mm catheter had an at least twofold greater odds of thrombus, charring, and bubbling, but larger lesions than the 10 mm multitemperature sensor catheter.

CONCLUSIONS

Catheter type, contact conditions, and power settings all play a role in lesion size and in the frequency of complications that occur during an RFA. Cooling the electrode tip, either internally or externally, does not prevent complications from occurring, especially at the higher power control settings. Adding more temperature sensors to the 10 mm seems to reduce the amount of complications that can occur.

ATP-sensitive potassium (K(+)(ATP)) channels regulate cell excitability and are expressed in steroid-responsive brain regions involved in sexual behavior, such as the preoptic area (POA) and medial basal hypothalamus (MBH). We hypothesized that K(+)(ATP) channels serve as a mechanism by which testosterone can control the electrical activity of neurons and consequently elicit male sexual responsiveness. RT-PCR analysis indicated that castration induces, while testosterone inhibits, mRNA expression of the K(+)(ATP) channel subunit Kir6.2 in both the POA and MBH of adult male rats. Intracerebral infusion of the pharmacological K(+)(ATP) channel inhibitor tolbutamide increased the proportion of long-term castrates displaying sexual behavior and restored mount frequency, intromission frequency, and copulatory efficacy to values observed in testes-intact animals. Infusions of tolbutamide, but not vehicle, also decreased latencies to mount and intromit in castrated males. Unilateral tolbutamide infusion directly into the POA significantly reduced mount latency of castrates; however, it did not affect other copulatory measures, suggesting that blockade of K(+)(ATP) channels in additional brain regions may be necessary to recover the full range of sexual behavior. These data indicate that blockade of K(+)(ATP) channels is sufficient to elicit the male sexual response in the absence of testosterone. Our observations are consistent with the hypothesis that testosterone modulates male sexual behavior by regulating K(+)(ATP) channels in the brain. Decreased channel expression or channel blockade may increase the excitability of androgen-target neurons, rendering them more sensitive to the hormonal, chemical, and somatosensory inputs they receive, and potentially increase secretion of neurotransmitters that facilitate sexual behavior.

The oral mucosa is an attractive cell source for autologous transplantation in human patients who require regenerative therapies of various epithelia. However, the time-course of cellular changes in transplanted oral mucosal epithelia at ectopic sites remains poorly understood. By applying a rat model, we analyzed phenotypic changes in oral mucosal epithelial cell sheets after harvest from temperature-responsive culture dishes and subsequent autologous subcutaneous transplantation. We used monoclonal antibodies to identify epithelial-specific cytokeratins 4, 10, 13, and 14, the stem/progenitor cell marker p63, and proliferating cell nuclear antigen, within the regenerated tissues. Transplanted oral mucosal epithelial cell sheets proliferated during the first week after grafting in conjunction with host inflammation, but then began to degenerate afterward with complete disappearance after 3 weeks. Our findings suggest that host subcutaneous tissues support proliferation and differentiation of the oral mucosal epithelial cell sheets, but are unable to promote maintenance of stem and progenitor cells and therefore cannot produce long-term survivability.