Isolation of Positive Modulator of Glucagon-like Peptide-1 Signaling from Trigonella foenum-graecum (Fenugreek) Seed.

The glucagon-like peptide-1 receptor (GLP-1R) is expressed in many tissues and has been implicated in diverse physiological functions, such as energy homeostasis and cognition. GLP-1 analogs are approved for treatment of type 2 diabetes and are undergoing clinical trials for other disorders, including neurodegenerative diseases. GLP-1 analog therapies maintain chronically high plasma levels of the analog and can lead to loss of spatiotemporal control of GLP-1R activation. To avoid adverse effects associated with current therapies, we characterized positive modulators of GLP-1R signaling. We screened extracts from edible plants using an intracellular cAMP biosensor and GLP-1R endocytosis assays. Ethanol extracts from fenugreek seeds enhanced GLP-1 signaling. These seeds have previously been found to reduce glucose and glycated hemoglobin levels in humans. An active compound (N55) with a new N-linoleoyl-2-amino-γ-butyrolactone structure was purified from fenugreek seeds. N55 promoted GLP-1-dependent cAMP production and GLP-1R endocytosis in a dose-dependent and saturable manner. N55 specifically enhanced GLP-1 potency more than 40-fold, but not that of exendin 4, to stimulate cAMP production. In contrast to the current allosteric modulators that bind to GLP-1R, N55 binds to GLP-1 peptide and facilitates trypsin-mediated GLP-1 inactivation. These findings identify a new class of modulators of GLP-1R signaling and suggest that GLP-1 might be a viable target for drug discovery. Our results also highlight a feasible approach for screening bioactive activity of plant extracts.

Genetic alterations of KRAS and TP53 in intrahepatic cholangiocarcinoma associated with poor prognosis.

The aim of this study is to investigate certain genetic features of intrahepatic cholangiocarcinoma (ICCA). A total of 12 eligible ICCA patients were enrolled, and tumor tissues from the patients were subjected to next-generation sequencing of a multi-genes panel. Tumor mutation burden (TMB), mutated genes, copy number variants (CNVs), and pathway enrichment analysis were performed. The median TMB was 2.76 Mutation/Mb (range, 0-36.62 Mutation/Mb) in ICCA patients. The top two most commonly mutated genes in ICCA were KRAS (33%) and TP53 (25%). The co-mutations of KRAS and TP53 were 16.7% (2/12) in ICCA patients. Notably, patient P6 with the highest TMB did not have KRAS and TP53 mutations. Additionally, TP53 and/or KRAS alterations were significantly associated with poor progression-free survival than those with wild type (1.4 months vs 18 months). DNA damage repair and homologs recombinant repair deficiencies were significantly associated with high TMB in ICCA cases. In conclusion, we found that certain genetic mutations of TP53 and KRAS could predict poor prognosis in ICCA patients.

Extension of shelf life of semi-dry longan pulp with gaseous chlorine dioxide generating film.

A packaging system using gaseous chlorine dioxide generating film (CDGF) in a sealed container was developed to extend the shelf life of semi-dry longan pulp (moisture content 38.8 wt%; aw0.8). The antimicrobial properties, formation of chloroxyanion residues and effects of CDGF on the quality of semi-dry longan pulp were investigated. CDGF was triggered by the moisture vapor from semi-dry longan pulp in the sealed container and released gaseous ClO2 into the headspace of the container. The antifungal test showed that CDGF significantly inactivated artificially inoculated molds in semi-dry longan pulp and achieved reductions of over 3 log CFU/g after 28 days storage at room temperature (25 °C). CDGF reduced total aerobic bacterial populations by over 6.4 log CFU/g and maintained these population levels at around 2.0 log CFU/g throughout the 180-day storage period at room temperature. The residual concentrations of chloride, chlorate and perchlorate in longan pulp increased and then decreased during the 180-day storage. Residual chloride levels were maintained at 1.5 mg/g after Day 120 and residual chlorate and perchlorate levels were not detected after Day 120 and Day 180, respectively, in CDGF-treated samples. CDGF treatments reduced total polyphenol content but didn't have any significant impact on the levels of polysaccharides in samples. There were no significant differences between CDGF-treated and control samples in color changes during storage. The content of 5-hydroymethylfurfural (5-HMF) in both samples increased during storage, suggesting that the Maillard reaction occurred. This study demonstrated an effective approach to develop a new antimicrobial packaging system for semi-dry longan pulp.

miR-140-3p inhibits breast cancer proliferation and migration by directly regulating the expression of tripartite motif 28.

The present study aimed to determine the expression profile and significance of microRNA-140-3p (miR-140-3p) in breast cancer (BC). miR-140-3p expression in BC tumor tissues and cell lines was measured by reverse transcription-quantitative polymerase chain reaction. Luciferase activity reporter assay and western blotting were used to assess the effect of miR-140-3p expression on tripartite motif 28 (TRIM28). Cell growth, migration, and invasion were assessed by Cell Counting Kit-8 assay, wound-healing assay and Transwell invasion assay, respectively. miR-140-3p expression was significantly reduced in BC tumor tissues compared with in adjacent normal tissues. Additionally, low miR-140-3p expression was found to predict poor prognosis of patients with BC. TRIM28 expression was significantly reduced by miR-140-3p overexpression in BC cell lines, and was inversely correlated with miR-140-3p in BC tissues. Overexpression of miR-140-3p also inhibited cell proliferation, migration and invasion compared with in the control group. In conclusion, the present study revealed that miR-140-3p inhibited the progression of BC partially by regulating TRIM28.