d-Allulose Supplementation Prevents Diet-Induced Hepatic Lipid Accumulation via miR-130-Mediated Regulation in C57BL/6 Mice.

SCOPE: d-allulose is a low-calorie rare sugar. It has been reported that d-allulose supplementation significantly inhibits diet-induced hepatic fat accumulation. However, the underlying molecular mechanisms remain unclear. This study elucidates the mechanism underlying the suppressive effect of d-allulose on hepatic fat accumulation in terms of miRNA regulation. METHODS AND RESULTS: Male C57BL/6 mice are divided into three experimental groups-normal diet and distilled water (CC group), high-fat diet (HFD) and distilled water (HC group), and HFD and 5% d-allulose solution (HA group)-and fed the respective diets for 8 weeks. Weight gain is significantly lower in the HA group than that in the HC group, although the caloric intake is the same in both. Histological analysis of liver tissues reveals excessive lipid accumulation in the HC group; this is greatly attenuated in the HA group. Real-time PCR and western blot analyses demonstrate that, compared to the HC group, the HA group exhibits decreased hepatic PPARγ and CD36 expression. Hepatic miR-130 expression levels are higher in the HA group than those in the CC and HC groups. CONCLUSIONS: These results indicate that miRNA changes associated with PPARγ may underlie the suppression of hepatic lipid accumulation induced by d-allulose intake.

Pyrosequencing assay to measure LINE-1 methylation level in esophageal squamous cell carcinoma.

BACKGROUND: Genome-wide DNA hypomethylation plays a role in genomic instability and carcinogenesis. DNA methylation in the long interspersed nucleotide element 1 L1 (LINE-1) repetitive element is a good indicator of global DNA methylation level. LINE-1 methylation is a useful marker for predicting cancer prognosis and monitoring efficacy of adjuvant therapy. Nonetheless, no study has examined LINE-1 methylation in esophageal squamous cell carcinoma (ESCC). The aim of this study is to assess the precision of sodium bisulfite conversion and polymerase chain reaction (PCR) pyrosequencing assay for evaluating LINE-1 methylation in ESCC. METHODS: To measure assay precision, we performed bisulfite conversion on 5 different DNA specimen aliquots (bisulfite-to-bisulfite) and repeated PCR pyrosequencing five times (run to run). Second, to assess heterogeneity of LINE-1 methylation levels within tumor, we made 5 different tissue sections from one tumor and examined LINE-1 methylation level of each section (section to section). Third, to evaluate LINE-1 methylation status in ESCC, we applied this assay to 30 ESCCs and 30 matched normal esophageal mucosa. RESULTS: Bisulfite-to-bisulfite standard deviation (SD) ranged from 1.44 to 2.90 (median 2.32) in ESCCs; and 0.57 to 4.02 (median 1.23) in normal esophagus. Run-to-run SD ranged from 0.63 to 3.25 (median 1.54) in ESCCs. Section-to-section SD ranged from 1.37 to 3.31 (median 1.94). ESCC tissues showed significantly lower levels of LINE-1 methylation than matched normal mucosa (P < .0001; n = 30). There was no significant relationship between LINE-1 methylation level and tumor stage (P = 0.14). CONCLUSIONS: Bisulfite conversion and PCR pyrosequencing assay can measure LINE-1 methylation in ESCC, and may be useful in clinical and research settings.

Use of 11C-methionine positron emission tomography in basal germinoma: assessment of treatment response and residual tumor.

INTRODUCTION: Prognosis of germinoma arising in the basal ganglia or thalamus is worse compared to that in the pineal or suprasellar region. One of the reasons for poor prognosis is the difficulty in evaluating the efficacy of treatment by conventional neuroimaging tools. PET STUDIES: The usefulness of (11)C-methionine (MET) positron emission tomography (PET) in monitoring the biological nature of brain tumors has been proved in glioma patients. CASE REPORTS: Herein, we describe MET-PET findings in three cases of germinomas in the basal ganglia or thalamus and discuss the use of MET-PET in the assessment of treatment response and residual tumor for the next treatment strategy. The patients showed transient increase of MET uptake in the lesions after the initial treatment. Although we did not perform histological verification, MET- PET findings suggested that active tumor cells were still alive in the lesions after the initial treatment. MET uptake gradually decreased during the course of intensive therapy in these patients. MET-PET also revealed germinoma invasion in the brain before discernible signal abnormality or mass lesion in conventional magnetic resonance images in two patients. DISCUSSION: Further studies including histological verification and long-term follow-up might validate the use of MET-PET in monitoring the treatment efficacy and evaluation of active residual tumor after the treatment. CONCLUSION: Until we understand what MET uptake truly represents, treatment strategy based on MET uptake must be carefully designed to prevent overtreatment and resultant complications.