a b s t r a c tInduced pluripotent stem (iPS) cells were recently established from human fibroblasts. In the present study we investigated the adipogenic differentiation properties of four human iPS cell lines and compared them with those of two human embryonic stem (ES) cell lines. After 12 days of embryoid body formation and an additional 10 days of differentiation on Poly-L-ornithine and fibronectincoated dishes with adipogenic differentiation medium, human iPS cells exhibited lipid accumulation and transcription of adipogenesis-related molecules such as C/EBPa, PPARc2, leptin and aP2. These results demonstrate that human iPS cells have an adipogenic potential comparable to human ES cells.

Objective: To elucidate early change of intra-abdominal fat in response to calorie restriction in patients with obesity by weekly evaluation using a dual bioelectrical impedance analysis (Dual BIA) instrument. Design and Methods: For 67 Japanese patients with obesity, diabetes, or metabolic syndrome, intraabdominal fat area (IAFA), initially with both Dual BIA and computed tomography (CT), and in subsequent weeks of calorie restriction, with Dual BIA were measured. Results: IAFA by Dual BIA (Dual BIA-IAFA) correlated well with IAFA by CT (CT-IAFA) in obese patients (r ¼ 0.821, P < .0001, n ¼ 67). Ten males and 9 females (age 49.0 6 14.4 years, BMI 33.2 6 7.3 kg/m 2 ) lost more than 5% of baseline body weight (BW) in 3 weeks, and their Dual BIA-IAFA, BW, and WC decreased by 18.9%, 5.3%, and 3.8%, respectively (P < .05, ANCOVA). Conclusion: Dual BIA instrument could detect the weekly change of Dual BIA-IAFA under calorie restriction in obese patients and demonstrated a substantially larger change of IAFA compared with changes of BW and WC in early weeks. This observation corroborates the significance of evaluating IAFA as a biomarker for obesity, and indicates the clinical usefulness of the Dual BIA instrument.

Aims/hypothesis The aim of this study was to generate induced pluripotent stem (iPS) cells from patients with mitochondrial DNA (mtDNA) mutation. Methods Skin biopsies were obtained from two diabetic patients with mtDNA A3243G mutation. The fibroblasts thus obtained were infected with retroviruses encoding OCT4 (also known as POU5F1), SOX2, c-MYC (also known as MYC) and KLF4. The stem cell characteristics were investigated and the mtDNA mutation frequencies evaluated by Invader assay.Results From the two diabetic patients we isolated four and ten putative mitochondrial disease-specific iPS (Mt-iPS) clones, respectively. Mt-iPS cells were cytogenetically normal and positive for alkaline phosphatase activity, with the pluripotent stem cell markers being detectable by immunocytochemistry. The cytosine guanine dinucleotide islands in the promoter regions of OCT4 and NANOG were highly unmethylated, indicating epigenetic reprogramming to pluripotency. Mt-iPS clones were able to differentiate into derivatives of all three germ layers in vitro and in vivo. The Mt-iPS cells exhibited a bimodal degree of mutation heteroplasmy. The mutation frequencies decreased to an undetectable level in six of 14 clones, while the others showed several-fold increases in mutation frequencies (51-87%) compared with those in the original fibroblasts (18-24%). During serial cell culture passage and after differentiation, no recurrence of the mutation or no significant changes in the levels of heteroplasmy were seen. Conclusions/interpretation iPS cells were successfully generated from patients with the mtDNA A3243G mutation. Mutation-rich, stable Mt-iPS cells may be a suitable source of cells for human mitochondrial disease modelling in vitro. Mutation-free iPS cells could provide an unlimited, diseasefree supply of cells for autologous transplantation therapy.

Five components of neutral horseradish peroxidase were isolated and purified by means of column chromatography, and designated B1, B2, B3, C1, and C2, respectively. All the components contained 2 atoms of calcium and 16.8-to-21.0% as much carbohydrate as in the enzyme molecule. They were very similar to one another with respect to physicochemical and chemical properties such as molecular weight, molar absorption coefficient, rate constants of the catalytic reaction and dissociation of cyanide compound, but were dissimilar with respect to isoelectric point. Values of the isoelectric points determined from column isoelectric focusing at 20 degrees C were 5.75 (B1), 7.15 (B2), 7.10 (B3), 9.40 (C1), and 9.63 (C2). However, these values varied significantly depending upon the method and conditions of the focusing. The acid-alkaline titration curves of components B2 and C1 were flat in the pH region of 6 to 9. The facts suggest that a slight difference in the number of ionized groups of the components causes a large difference in the isoelectric points.

Notch signaling regulates cell fate determination in various tissues. We have reported the generation of mice with a pancreas-specific knockout of Rbp-j using Pdx.cre mice. Those mice exhibited premature endocrine and ductal differentiation. We now generated mice in which the Rbp-j gene was inactivated in Ptf1a-expressing cells using Ptf1a.cre mice. The timing of the Cre-mediated deletion in Rbp-j f/f Ptf1a.cre mice is 1 day later than that in Rbp-j f/f Pdx.cre mice. In Rbp-j f/f Ptf1a.cre mouse pancreases, at E13.5, the reduced Hes1 expression was accompanied by reduced epithelial growth, but premature endocrine cell differentiation was minimal. At E15.5, Pdx1 expression was repressed and acinar cell differentiation was reduced, but an increase in acinar cell proliferation was observed during the perinatal period. Our study indicates that, in addition to its role in preventing premature differentiation of early endocrine cells, Rbp-j regulates epithelial growth, Pdx1 expression, and acinar cell differentiation during mid-pancreatic development. Developmental Dynamics 236:2779 -2791, 2007.