Application of Digital Image Analysis to Determine Pancreatic Islet Mass and Purity in Clinical Islet Isolation and Transplantation.

Pancreatic islet mass, represented by islet equivalent (IEQ), is the most important parameter in decision making for clinical islet transplantation. To obtain IEQ, the sample of islets is routinely counted under a microscope and discarded thereafter. Islet purity, another parameter in islet processing, is routinely assessed by estimation only. In this study, we validated our digital image analysis (DIA) system by using the software of Image Pro Plus and a custom-designed Excel template to assess islet mass and purity to better comply with current good manufacturing practice (cGMP) standards. Human islet samples (60 collected from a single isolation and 24 collected from 12 isolations) were captured as calibrated digital images for the permanent record. Seven trained technicians participated in determination of IEQ and purity by the manual counting method (manual image counting, Manual I) and DIA. IEQ count showed statistically significant correlations between the Manual I and DIA in all sample comparisons (r > 0.819 and p < 0.0001). A statistically significant difference in IEQ between Manual I and DIA was not found in all sample groups (p > 0.05). In terms of purity determination, statistically significant differences between assessment and DIA measurement were found in high-purity 100-µl samples (p < 0.005) and low-purity 100-µl samples (p < 0.001) of the single isolation. In addition, islet particle number (IPN) and the IEQ/IPN ratio did not differ statistically between Manual I and DIA. In conclusion, the DIA used in this study is a reliable technique to determine IEQ and purity. Islet sample preserved as a digital image and results produced by DIA can be permanently stored for verification, technical training, and information exchange among islet centers. Therefore, DIA complies better with cGMP requirements than the manual counting method. We propose DIA as a quality control tool to supplement the established standard manual method for islet counting and purity estimation.

Bariatric surgery to treat obesity.

Obesity has become a global health problem that contributes to numerous life-threatening and disabling diseases, such as type 2 diabetes mellitus and coronary artery disease. The long-term results of traditional weight loss therapies, including diet, exercise, and medications, are relatively poor. Bariatric surgery is the most effective treatment of morbidly obese patients to allow substantial, sustained weight loss and to improve or resolve obesity-associated comorbidities, thereby reducing mortality. According to US practice guidelines, patients qualify for bariatric surgery with a body mass index of 35 kg/m(2) and associated comorbidities, or a body mass index of 40 kg/m(2), after failure of conservative weight loss measures. Currently, the established procedures in the United States are the laparoscopic Roux-en-Y gastric bypass, adjustable gastric banding, sleeve gastrectomy, and biliopancreatic diversion with duodenal switch. The surgeries vary substantially in their postoperative amount of weight loss, resolution of comorbidities, nutritional requirements, and nature and severity of complications. There is no perfect bariatric surgery, an informed risk and benefit assessment should be made by each patient. The procedures are safe, with a mortality rate of 0.3%. Sustained weight loss depends finally on patient re-education in terms of diet, need for regular exercise, and careful bariatric follow-up evaluation.

Glucose and lipid metabolism after liver transplantation in inbred rats: consequences of hepatic denervation.

The liver plays a central role in glucose and lipid homeostasis. Because liver transplantation severs the hepatic nerves which influence this function, we hypothesized that insulin resistance and hyperlipidemia develop after liver transplantation, thus increasing the atherosclerotic risk. Therefore, we studied inbred rats 8 months after orthotopic liver transplantation (Tx, n = 39) or laparotomy (sham, n = 37) by either oral glucose tolerance test (Tx, n = 13; sham, n = 8), meal tolerance test (Tx, n = 9; sham, n = 13), or euglycemic hyperinsulinemic clamp with tritiated glucose infusion (Tx, n = 17; sham, n = 16). We found that liver transplantation significantly increased basal hepatic glucose production (HGP) in the clamp study by 20% (37.3 +/- 2.2 vs 31.0 +/- 2.1 micromol kg -1 .min -1 , P < .05) and fasting plasma low-density lipoprotein (LDL) cholesterol by 36% (0.79 +/- 0.06 vs 0.58 +/- 0.05 mmol/L, P < .05). However, it did not affect HGP, total glucose uptake, metabolic clearance rate of insulin, and suppression of plasma nonesterified fatty acids, which were all normal in response to rising plasma insulin concentrations in the dose-response clamp studies. The oral glucose tolerance test and meal tolerance test also showed normal glucose and nonesterified fatty acids homeostasis with adequate pancreatic insulin secretion and hepatic insulin clearance after liver transplantation. The only consequences of liver transplantation are increased basal HGP and plasma LDL cholesterol, which may be caused by persistent vagal denervation of the liver. Although insulin resistance is absent, elevated plasma LDL cholesterol increases the atherosclerotic risk.