Individuals with Type 2 Diabetes Mellitus Tend to Select Low-Carbohydrate, Low-Calorie Food Menus at Home on Diet Application

(1) Background: From the perspective of patient-centered care, it is important for medical professionals involved in diabetes care to know the role of choice behavior when individuals with type 2 diabetes mellitus select their meals at home. In Japan, online meal management applications are widely used to help individuals to prepare healthy, colorful, and tasty meals. (2) Objective: To assess menu selection from an online diet management application in individuals with type 2 diabetes mellitus over a period of 24 months. (3) Method: The saved data of the selected food menus on the online diet management application were analyzed. We identified specific nutritional groups of the food menus, called nutritional clusters, by clustering the multidimensional data of the nutrients after de-dimensioning them. Then, we analyzed the constitutional nutrients of each nutritional cluster with the highest and lowest frequencies of selection by the users of the application. (4) Results: In all, 9674 food menus made by 3164 people were included in the analysis, and 12 nutritional clusters were identified. Low-carbohydrate and low-calorie food clusters showed the highest selection frequency. The average caloric value of 149.7 kcal and average carbohydrate ratio of 47% in the cluster with the highest selection frequency were significantly lower than the average caloric value of 435.2 kcal and carbohydrate ratio of 63% in the cluster with the lowest selection frequency (p < 0.001, respectively). (5) Conclusion: Individuals with type 2 diabetes in this population preferred to select lower-carbohydrate and lower-calorie food menus at home using online diet management applications. To improve sustained self-management and quality of life, medical professionals may consider incorporating preferred dietary behaviors into medical management of type 2 diabetes mellitus

Augmented Growth Hormone Secretion and Stat3 Phosphorylation in an Aryl Hydrocarbon Receptor Interacting Protein (AIP)-Disrupted Somatotroph Cell Line

<div><p>Aryl hydrocarbon receptor interacting protein (<i>AIP</i>) is thought to be a tumor suppressor gene, as indicated by a mutational analysis of pituitary somatotroph adenomas. However, the physiological significance of <i>AIP</i> inactivation in somatotroph cells remains unclear. Using CRISPR/Cas9, we identified a GH3 cell clone (termed GH3-FTY) in which <i>Aip</i> was genetically disrupted, and subsequently investigated its character with respect to growth hormone (Gh) synthesis and proliferation. Compared with GH3, GH3-FTY cells showed remarkably increased Gh production and a slight increase in cell proliferation. Gh-induced Stat3 phosphorylation is known to be a mechanism of Gh oversecretion in GH3. Interestingly, phosphorylated-Stat3 expression in GH3-FTY cells was increased more compared with GH3 cells, suggesting a stronger drive for this mechanism in GH3-FTY. The phenotypes of GH3-FTY concerning Gh overproduction, cell proliferation, and increased Stat3 phosphorylation were significantly reversed by the exogenous expression of <i>Aip</i>. GH3-FTY cells were less sensitive to somatostatin than GH3 cells in the suppression of cell proliferation, which might be associated with the reduced expression of somatostatin receptor type 2. GH3-FTY xenografts in BALB/c nude mice (GH3-FTY mice) formed more mitotic somatotroph tumors than GH3 xenografts (GH3 mice), as also evidenced by increased Ki67 scores. GH3-FTY mice were also much larger and had significantly higher plasma Gh levels than GH3 mice. Furthermore, GH3-FTY mice showed relative insulin resistance compared with GH3 mice. In conclusion, we established a somatotroph cell line, GH3-FTY, which possessed prominent Gh secretion and mitotic features associated with the disruption of <i>Aip</i>.</p></div

Selective androgen receptor modulator, S42 has anabolic and anti-catabolic effects on cultured myotubes

We previously identified a novel selective androgen receptor modulator, S42, that does not stimulate prostate growth but has a beneficial effect on lipid metabolism. S42 also increased muscle weight of the levator ani in orchiectomized Sprague–Dawley rats. These findings prompted us to investigate whether S42 has a direct effect on cultured C2C12 myotubes. S42 significantly lowered expression levels of the skeletal muscle ubiquitin ligase (muscle atrophy-related gene), atrogin1 and Muscle RING-Finger Protein 1(MuRF1) in C2C12 myotubes, as determined by real time PCR. Phosphorylation of p70 S6 kinase (p70S6K), an essential factor for promoting protein synthesis in skeletal muscle, was significantly increased by S42 to almost the same extent as by insulin, but this was significantly prevented by treatment with rapamycin, an inhibitor of mechanistic target of rapamycin complex 1 (mTORC1). However, phosphorylation of Akt, upstream regulator of mTORC1, was not changed by S42. S42 did not increase insulin-like growth factor 1 (Igf1) mRNA levels in C2C12 myotubes. These results suggest that S42 may have an anabolic effect through activation of mTORC1–p70S6K signaling, independent of IGF-1-Akt signaling and may exert an anti-catabolic effect through inhibition of the degradation pathway in cultured C2C12 myotubes. Keywords: SARM, C2C12, Myotube, Muscl

Screening of <i>Aip</i>-disrupted GH3 clones generated by CRISPR/Cas9.

<p>The detailed mutations of clones 1 and 2 are described in the Results section and <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0164131#pone.0164131.s001" target="_blank">S1 Fig</a></b>. Clone 1 corresponds to GH3-FTY cells intensively investigated in the present study. (<b>A</b>) Western blot analysis of Aip protein in GH3 and mutant GH3 clones (clones 1 and 2). Fifteen μg of protein from cell lysates was subjected to SDS-PAGE and immunoblotted with antibodies against Aip and beta-actin. Each target protein was visualized using VersaDoc 5000 (BIO-RAD, Tokyo, Japan). (<b>B, C</b>) The relative expression of <i>Gh</i> mRNA and <i>Prl</i> mRNA to <i>Actb</i> mRNA is shown, respectively. Data were compared using the unpaired two-tailed <i>t</i>-test. **<i>P</i><0.01 (clone 1 or 2) vs GH3. (<b>D)</b> The relative expression of <i>Aip</i> mRNA to <i>Actb mRNA</i> in cultured clone 1 (GH3-FTY cells) and clone 2 is shown. Data were compared using the unpaired two-tailed <i>t</i>-test. * <i>P</i><0.05 (clone 1 or 2) vs GH3.</p

Plasma Gh and Igf-1 levels in GH3 and GH3-FTY mice.

<p>BALB/c-nu mice were inoculated with GH3 or GH3-FTY cells. Control mice were inoculated by the medium only. Plasma levels of Gh (<b>A</b>) and Igf-1 (<b>B</b>) at 8 weeks after inoculation. Data were compared using the one-way ANOVA with Tukey’s post-hoc test. *<i>P</i><0.05, ***<i>P</i><0.001. ns, not significant.</p

Gh secretion in the cultured medium, <i>Gh</i> mRNA levels before and after exogenous <i>Aip</i> expression, and intracellular cAMP content in cultured GH3 and GH3-FTY cells.

<p>(<b>A</b>) Gh secretion in the media from cultured GH3 and GH3-FTY cells was measured and expressed as a fold increase. (<b>B</b>) <i>Gh</i> mRNA levels in cultured GH3 and GH3-GTY cells were calculated relative to <i>Actb</i> (internal control) using the ∆Ct method. (<b>C</b>) The effect of lentivirus (LV)-mediated forced expression of exogenous <i>gfp</i> as a control or <i>Aip</i> on <i>Gh</i> mRNA levels. GFP and Aip indicate LV-GFP and LV-Aip, respectively. (<b>D</b>) The intracellular cAMP content of both cultured cells was measured as described in the Methods. Data were compared using the unpaired two-tailed <i>t</i>-test. **<i>P</i><0.01. ns, not significant.</p

Western blot analysis of p-Stat3 (Tyr705) and Stat3 in cultured GH3 and GH3-FTY cells.

<p>(<b>A</b>) Western blot analysis of p-Stat3 (Tyr705) and Stat3. Twenty μg protein from cell lysates was separated by SDS-PAGE and immunoblotted with antibodies against p-Stat3, Stat3, and beta-actin. (<b>B</b>) Statistical evaluation of Stat3/ beta-actin expression between cultured GH3 and GH3-FTY cells. Data were compared using the unpaired two-tailed <i>t</i>-test. ns, not significant. The intensity of each detected band was analyzed using the image analysis software Quantity One (BIO-RAD), and the Stat3/ beta-actin ratio was calculated. (<b>C</b>) Statistical evaluation of p-Stat3/ beta-actin expression between cultured GH3 and GH3-FTY cells. Data were compared using the unpaired two-tailed <i>t</i>-test. **<i>P</i><0.01. (<b>D)</b> Western blot analysis of p-Stat3 and Stat3. The effect of lentivirus (LV)-mediated forced expression of exogenous <i>Aip</i> (LV-AIP) or <i>gfp</i> (LV-GFP) as a control on p-Stat3 expression was examined. Twenty μg protein from cell lysates was subjected to SDS-PAGE and immunoblotted with antibodies against p-Stat3 and beta-actin. (<b>E)</b> Statistical evaluation of p-Stat3. GFP and Aip indicate LV-GFP and LV-Aip, respectively. Data were compared using the unpaired two-tailed <i>t</i>-test. **<i>P</i><0.01. ns, not significant. (<b>F</b>) <i>Il6r</i> mRNA levels relative to <i>Actb</i> mRNA levels as determined by qPCR. (<b>G</b>) <i>Il6r</i> mRNA levels relative to <i>Actb</i> mRNA levels as determined by qPCR before and after lentivirus (LV)-mediated exogenous expression of <i>Aip</i> (LV-Aip) or <i>gfp</i> (LV-GFP) as a control. GFP and Aip indicate LV-GFP and LV-Aip, respectively. Data were compared using the unpaired two-tailed <i>t</i>-test. *<i>P</i><0.05, **<i>P</i><0.01. ns, not significant.</p