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Thyroid hormones are critical for the development of opsins involved in color vision. Hypothyroid mice show delayed M-opsin development and expanded distribution of S-opsin on the retina. However, the effects of maternal hypothyroidism on opsin development remain unknown. This study investigates the effects of congenital central hypothyroidism and maternal hypothyroidism on opsin development in thyrotropin-releasing hormone knockout (TRH−/−) mice. We examined the mRNA expression and protein distribution of S/M-opsin on postnatal days (P)12 and 17, as well as mRNA expression of type 2 and 3 iodothyronine deiodinase (DIO2 and DIO3, respectively) in the retina and type 1 iodothyronine deiodinase (DIO1) in the liver at P12 in TRH+/− mice born to TRH+/− or TRH−/− dams, and conducted S/M-opsin analysis in TRH+/+ or TRH−/− mice born to TRH+/− dams at P12, P17, and P30. M-opsin expression was lower in TRH+/− mice born to TRH−/− dams than in those born to TRH+/− dams, whereas S-opsin expression did not significantly differ between them. DIO1, DIO2, and DIO3 mRNA expression levels were not significantly different between the two groups; therefore, thyroid function in peripheral tissues in the pups was similar. S/M-opsin expression did not significantly differ between the TRH+/+ and TRH−/− mice born to TRH+/− dams on any postnatal day. These results demonstrate that maternal hypothyroidism causes M-opsin developmental delay during the early developmental stages of neonatal mice, and TRH−/− mice, a model of congenital central hypothyroidism, born to a euthyroid dam do not have delayed opsin development.

This study aimed to investigate whether persistently high 1-h postchallenge glucose (PG) levels in a 75 g oral glucose tolerance test (75 g OGTT), as well as persistently low 1-h PG levels, are a risk factor for reclassification from normal glucose tolerance (NGT) into impaired glucose tolerance (IGT) and type 2 diabetes mellitus (T2DM) among participants continually observed for 11 years. This single-center retrospective study used the electronic records of all participants undergoing Ningen Dock (health checkup) at Kiryu Kosei General Hospital between 2008 and 2018. In 2008, 361 of 523 participants who received Ningen Dock had NGT. Of the 361 participants, 109 received 75 g OGTT yearly for 11 years (2008–2018), and 72 of these 109 participants showed either persistently high 1-h PG (> 155 mg/dL) or persistently low 1-h PG (

Resolvins are pro-resolving lipid mediators with highly potent anti-inflammatory effects. Because of their polyunsaturated structures, however, they are unstable to oxygen as a drug prototype. To address this issue, we designed and synthesized CP-RvE3 as oxidatively stable congeners of RvE3 by replacing the

Background: Previously, AF-956, which contains S356 of FAM83G and an N-terminal antenna peptide for entry into colon cancer cells, is markedly antiproliferative compared to a control peptide (AF-859), which lacks the N-terminal antenna peptide, by inducing apoptosis via the inhibition of HSP27 phosphorylation at residues S15 and S82. Objective: Because FAM83G-derived peptides are promising lead compounds for colon cancer treatment, we reanalyzed the effect of AG-066, which contains S356 of FAM83G and an N-terminal antenna peptide for entry into the liver cancer cells. Methods: HepG2 liver cancer cells were incubated with either AF-859 or AG-066 at a concentration of 54 μM at 37 °C for 24, 48, and 72 h. The effects of AF-859 and AG-066 on the cultured HepG2 cells were estimated using an inverted light microscope. Furthermore, the DNA ladder method and the dead cell assay were performed by applying Live/Dead Cell Staining Kit II. Erk phosphorylation was estimated by western blotting. Results: Treatment with AG-066 markedly reduced HepG2 viable cell counts compared to the AF- 859-treated HepG2 cells, as evident from the significantly increased number of dead cells in the culture medium. Additionally, AG-066 treatment increased cellular DNA laddering. We found no difference in Erk phosphorylation status between the AG-066- and AF-859-treated groups. Conclusion: This study illustrated that the peptide with a structure based on FAM83G functions as a spontaneous apoptosis inducer for liver cancer cells. Hence, it is a promising lead compound for the treatment of liver cancer.

Exocyst complex component 3-like 1 (EXOC3L1), which regulates insulin secretion, is ubiquitously present in heart, lung, liver, spleen, kidney, muscle, cerebellum, pituitary, adrenal grand, and pancreatic islets. Its deduced amino acid sequence has 31% identity and 53% similarity with Sec6, so they are considered isoforms. Since Sec6 suppresses apoptosis via HSP27, we investigated the involvement of EXOC3L1 expression in apoptosis. We found that overexpressed EXOC3L1 in Chinese hamster ovary cells significantly reduced cultured cell numbers. It also significantly increased apoptotic DNA ladder, caspase 3 activity, and cleavage of caspase 3 compared with the control. Thus, although Sec6 reduces apoptosis by increasing HSP27 phosphorylation, overexpressed EXOC3L1 alone can spontaneously induce apoptosis without apoptotic stimulators or inducers.

Objective: Because patients with diabetes mellitus (DM) were forced to stay indoors during the state of emergency, resulting in stress and a lack of physical activity, concerns about their glycemic control were raised. Patients and Methods: The 165 patients' glycated hemoglobin (HbA1c) levels were compared during the following periods: the 4 months that were selected as a representative condition 1 year before the COVID-19 pandemic (May 2018, March 2019, June 2019, and July 2019) and the latter 3 months as a 1-year follow-up during the COVID-19 pandemic (May 2019, March 2020, June 2020, and July 2020). Results: The patients' HbA1c levels were 7.32 ± 1.23, 7.44 ± 1.20, 7.16 ± 1.06, 7.01 ± 1.05, 7.23 ± 1.06, 7.45 ± 1.18, 7.15 ± 10.7, and 7.11 ± 1.17 in May 2018, March 2019, June 2019, July 2019, May 2019, March 2020, June 2020, and July 2020, respectively (expressed as mean ± standard deviation). Conclusion: The analysis showed that HbA1c levels did not worsen during the self-restraint period.

Triiodothyronine reduces sodium glucose cotransporter 2 expression in the kidney and increased glucose excretion in urine of alloxan-induced diabetic rats. Free thyroxine is also negatively associated with islet beta-cell function in euthyroid subjects. However, the influence of sodium glucose cotransporter 2 inhibitor on thyroid function in patients with type 2 diabetes mellitus has not been established.We investigated thyroid function in patients with type 2 diabetes mellitus in the presence or absence of sodium glucose cotransporter 2 inhibitor in a multicenter retrospective study conducted between 2019 and 2021. All participants visited the hospital monthly for type 2 diabetes mellitus treatment and plasma glucose and glycated hemoglobin level measurements. Furthermore, thyroid-stimulating hormone, free triiodothyronine, and free thyroxine levels were measured annually.Free triiodothyronine level and the free triiodothyronine:free thyroxine ratio in the group treated with sodium glucose cotransporter 2 inhibitor were significantly higher than the levels in the group not treated with sodium glucose cotransporter 2 inhibitor. Free triiodothyronine levels in the group treated with sodium glucose cotransporter 2 inhibitor were significantly higher than the levels in the group not treated with sodium glucose cotransporter 2 inhibitor (p = 0.040). Free thyroxine levels in the group treated with sodium glucose cotransporter 2 inhibitor were significantly lower than the levels in the group not treated with sodium glucose cotransporter 2 inhibitor (p = 0.002). Thyroid-stimulating hormone levels did not differ significantly between the two groups.Our findings show that sodium glucose cotransporter 2 inhibitor affects free triiodothyronine levels free thyroxine levels, and the free triiodothyronine:free thyroxine ratio.

In this study, we compared the efficacy of a dipeptidyl peptidase-4 inhibitor (DPP4i) to improve glucose control in patients with type 2 diabetes mellitus (T2DM) with or without Hashimoto's thyroiditis (HT). First, we compared the change in glycated hemoglobin (HbA1c) between the hypothyroid condition (before levothyroxine sodium hydrate [LT4] treatment) and euthyroid condition (after LT4 treatment when patients had achieved euthyroidism for at least six months) in patients with T2DM and HT. Next, we compared the change in HbA1c levels before and six months of DPP4i treatment in patients with T2DM with and without HT. In hypothyroid condition the change in HbA1c after six months of DPP4i treatment was 0.13% ± 0.86%. The change in HbA1c levels from when patients first achieved euthyroidism to after six months in the euthyroid condition was 0.26% ± 0.90%. DPP4i efficacy in patients with T2DM and HT was reduced compared to patients with T2DM but without HT (-0.40 ± 0.90 vs. -0.99 ± 0.5, p = 0.0032). These data suggest that hypothyroidism does not impact on DPP4i efficacy. However, the effect of DPP4i in patients with T2DM and HT was reduced compared to that in T2DM patients without HT. An estimation of thyroid function before prescribing DPP4i may be useful tool for predicting the efficacy of DPP4i, allowing the ruling out complications from HT.

Free triiodothyronine/free thyroxine (FT3/FT4) ratio is considered as an index of the activities of iodothyronine deiodinase types 1 and 2 (DIO1 and DIO2, respectively) and is reportedly associated with insulin resistance in euthyroid adults. Euthyroid women with polycystic ovary syndrome accompanied with insulin resistance have lesser deiodinase activities. Correspondingly, the serum insulin level in a fasted condition positively correlates with the FT3/FT4 ratio, and insulin depletion decreases the DIO2 activity in mice. Selected genetic variants in DIO1 are also associated with insulin resistance measures. Therefore, if insulin positively regulates DIO1 and DIO2, the FT3/FT4 ratio should decrease under impaired insulin action, and the casual insulin level and FT3/FT4 ratio should be negatively correlated. To evaluate this hypothesis, we conducted a single-center retrospective study between 2018 and 2021. All participants visited the selected hospitals monthly for type 2 diabetes mellitus treatment and casual plasma glucose and HbA1c level measurements. Furthermore, their casual serum insulin levels were measured annually. Meanwhile, we excluded patients treated with insulin injection. Ultimately, we evaluated 71 patients, which all exhibited euthyroid conditions. The FT3/FT4 ratio was independently associated with thyroid-stimulating hormone, casual plasma glucose, and casual insulin levels. In terms of the regression coefficients of the univariate linear regression analysis, the FT3/FT4 ratio negatively correlated with the casual serum insulin levels. Therefore, the risk of FT3/FT4 ratio underestimation should be considered when diagnosing Graves' disease, which is often accompanied with insulin resistance.

The Abbott FreeStyle Libre flash glucose monitoring system (FGM) is a recently introduced, but widespread continuous glucose monitoring system. While its mean absolute relative difference (MARD) value indicating its accuracy is acceptable with reference to the self-monitoring of blood glucose (SMBG) levels, few reports have examined the MARD in sensor glucose values of FGM (FGM-SG) with reference to plasma glucose (PG) levels and the factors determining it. We performed oral glucose tolerance tests (OGTTs) in 25 Japanese subjects without diabetes. Parkes error grid analyses showed that FGM-SG with either SMBG or PG levels as a reference met International Organization for Standardization criteria. The MARD in FGM-SG with reference to SMBG levels was 10.9 ± 4.1% during OGTTs. Surprisingly, the MARD in FGM-SG with reference to PG levels was 20.3 ± 10.3% during OGTTs, revealing a discrepancy in the accuracy of FGM-SG compared with that of PG levels; moreover, the MARD showed negative correlations with fasting blood sugar level, homeostasis model assessment insulin resistance index, and body mass index (BMI). Multiple regression analyses revealed that BMI contributed the most to the MARD when FGM-SG and PG level were compared, as lean individuals have a greater MARD regardless of glucose levels. Inaccurate FGM data could potentially increase the risk of inappropriate treatment; consideration of such factors is critical to ensure reliable FGM values.

Obesity-associated type 2 diabetes mellitus is associated with the development of insulin resistance. Among several metabolites, resolvins that are metabolites of eicosapentaenoic acid have been shown to exert insulin-sensitizing effects; however, the role of resolvin E3 (RvE3) in glucose metabolism has not been studied. In this study, the effect of RvE3 on glucose metabolism in mice with high-fat diet-induced obesity and 3T3L1 adipocytes was studied. C57BL/6 mice fed a high-fat diet were administered RvE3, for which insulin tolerance, oral glucose tolerance tests, and the homeostasis model assessment of insulin resistance, were performed. RvE3 treatment significantly improved insulin sensitivity and glucose tolerance and regulated protein kinase B (Akt) phosphorylation in the adipose tissue. Moreover, RvE3 treatment enhanced the insulin-stimulated glucose transporter 4 (Glut4) translocation, glucose uptake, phosphatidylinositol-3-kinase (PI3K) activity, and Akt phosphorylation in 3T3L1 adipocytes, whereas a PI3K inhibitor inhibited the enhanced insulin-stimulated glucose uptake induced by RvE3. These findings indicate that RvE3 likely improves insulin sensitivity, resulting in the upregulation of glucose uptake in adipocytes by activating the PI3K/Akt signaling pathways. Collectively, the findings of this study show that RvE3 may play a role in glucose homeostasis and could be used as a potential therapeutic target for developing treatments for obesity-associated diabetes.

Overexpressed TBC1D8B, a GTPase-activating protein, significantly reduced cultured HCT116 human colon cancer cell number. We tested N-terminal TBC1D8B, which is identical to wild type TBC1D8B from amino acid positions 1 to 427 and possesses a modified sequence from position 428 to 435 (ECGGLFLL) because of the introduction of a premature stop codon at position 436 to narrow down the minimum requirement element. The N-terminal TBC1D8B contains two GRAM domains but not the TBC domain essential for Rab-GTPase activity. The N-terminal TBC1D8B overexpression significantly reduced the cultured HCT116 cell number. When we tested C-terminal TBC1D8B, containing the portion of TBC1D8B absent in the N-terminal TBC1D8B, the cell number reduction was not observed. The N-terminal TBC1D8B overexpression significantly increased the coronin 1B expression and reduced the phosphorylation of serine 51 in eIF2α, respective markers of apoptosis and cell death/survival. Also, caspase 3 and poly ADP-ribose polymerase increased cleavage in suspended cells overexpressing the N-terminal TBC1D8B. Taken together, it is not the TBC domain for Rab-GTPase activity, but amino acids 1 to 435, including the two GRAM domains, that is enough for TBC1D8B to cause spontaneous apoptosis. TBC1D8B could be a potential anticancer therapeutic molecule.

Introduction Previously, we reported that the renal threshold for glucose reabsorption can be measured as the lowest plasma glucose level that correlates with the first detectable appearance of urine glucose. These data revealed significant variations among patients with type 2 diabetes mellitus (T2DM), and there was a significant negative correlation between the renal threshold for glucose reabsorption and HbA1c levels following treatment with the sodium-glucose co-transporter 2 (SGLT2) inhibitor ipragliflozin. Recently approved SGLT inhibitors may not show the same efficacy in patients with T1DM as in those with T2DM unless the renal threshold for glucose reabsorption shows similar levels between the two groups. SGLT2 inhibitors improve plasma glucose control in patients with T2DM by reducing glucose reabsorption via the epithelial cells of the proximal tubule. Methods The renal threshold for glucose reabsorption was defined as the minimum blood glucose concentration that results in the presence of measurable glycosuria in at least 12 measurements. Results The renal threshold for glucose reabsorption in patients with T2DM [n = 64; 201.8 ± 33.6 (range 121–268) mg/dL] was significantly higher than that in patients with T1DM [n = 33; 171.0 ± 33.0 (range 76–259) mg/dL; p = 0.00022]. Conclusion The renal threshold for glucose reabsorption in patients with T1DM was near the normal range and significantly lower than that in patients with T2DM. The efficacy of the SGLT2 inhibitor was better in patients with a higher renal threshold for glucose reabsorption. Thus, these results indicate that it is advisable to estimate the renal threshold for glucose reabsorption prior to initiating SGLT2 inhibitor therapy in patients with T1DM.

Context In patients with aldosterone-producing adenomas (APAs), adrenalectomy causes a rapid decrease in blood pressure and increase in blood potassium levels; however, the effects of these intensive metabolic changes on kidney function with age have not yet been examined in Japan. Objective To investigate factors related to the progression of kidney dysfunction after adrenalectomy in different age groups. Participants Fifty Japanese patients with APAs and 27,572 health checkup patients as controls were examined. Main Outcome Measures We investigated changes in estimated glomerular filtration rate (eGFR) after adrenalectomy and characterized patients who progressed to chronic kidney disease (CKD). Results The postoperative cutoff age of CKD is 50 years and age is a unique factor for the progression of CKD after adrenalectomy. Among preoperative patients, CKD was 6% for those

Anemia due to angiotensin receptor blocker (ARB) therapy has been previously reported in patients with diabetes mellitus with glomerular filtration rates of

Introduction: Retinal cone photoreceptor cells contain short (S) and medium (M) wavelength opsins, which are light-sensitive substances involved in color vision and visual acuity by sensing lights of different wavelengths. Thyroid hormones promote M-opsin expression and suppress S-opsin expression during the differentiation of cone photoreceptors. It was previously reported that M-opsin expression was delayed and S-opsin expression increased in TSH receptor-deficient mice and methimazole-induced hypothyroid mice. In addition, no M-opsin expression and increased S-opsin expression were observed in thyroid hormone receptor (TR) β2-deficient mice (Ng L et al, Nature Genetics. 2001; 27(1): 94-98.). This suggested that impaired thyroid function affects opsin development. We therefore examined retinal development in TRH-deficient mice, which are a model of central hypothyroidism established in our laboratory. Methods: We performed HE staining of the retina at postnatal 30 days and electroretinography at postnatal 10 weeks using TRH-/- and wild-type (WT) mice. We also examined expression levels of S/M opsin mRNA in WT, TRH-/- and TRH-/- pups born from TRH-/- dams at postnatal 12,17 and 30 days, and TRβΔ337T knock-in mice (TRβmut/mut) at postnatal 30 days. Furthermore, we performed immunohistochemistry to examine S/M opsin protein expression in these mice. Results: The retinal structures by HE staining and retinal functions by electroretinography in TRH-/- mice were unchanged compared with those in WT mice. Although M-opsin expression was not detected and S-opsin expression was higher in TRβmut/mut mice than in WT mice, the mRNA and protein expression levels of S/M-opsin did not significantly differ between TRH-/- pups born from TRH+/- dams and WT pups at all postnatal days. TRH-/- pups born from TRH-/- dams exposed to maternal hypothyroidism had similar serum total T4 levels to TRH-/- pups born from TRH+/- with normal maternal thyroid function. In contrast, the mRNA expression level of M-opsin was significantly lower (1.00±0.06 vs 0.64±0.05: mean ± SE, p

To evaluate retrospectively the satisfaction of continuous subcutaneous insulin infusion (CSII) compared to multiple daily infusions in Japanese women with diabetes mellitus (DM) during pregnancy, we examined 27 women with type 1 (n = 20) or type 2 (n = 6) diabetes mellitus or gestational diabetes (n = 1) who previously used CSII. Among these patients, 19 had used CSII during pregnancy, which accounted for 30 births. Questionnaires were retrospectively administered. The mean age of patients was 36.5 ± 7.0 years. The average birth weight was 3.1 ± 0.6 kg, with 62% of babies exhibiting complications. The satisfaction level of CSII for patients during pregnancy was 3.95 ± 0.26, whereas it was 1.84 ± 0.26 for multiple daily infusions (P

Previously, we reported that short-term continuous glucose monitoring (CGM) with the professional iPro2© CGM device is a good clinical indicator of glycated hemoglobin (HbA1c) levels. However, there was no significant correlation between CGM and HbA1c levels when HbA1c levels were >8.0%. To further investigate this issue, we performed a similar study using the FreeStyle Libre Pro©, a newer device that does not require glucose calibration and allows patients to be examined for up to 14 days. Fifty-nine patients (68% women, 32% men) were examined. Twenty-eight and 31 patients presented with type 1 and type 2 diabetes, respectively. Clinically assessed HbA1c levels were compared to blood glucose levels determined by the FreeStyle Libre Pro© for up to 14 days (10.7 ± 3.7 days). We found a significant correlation between HbA1c and CGM levels even when HbA1c levels were >8.0%. Additionally, the correlation between HbA1c and average glucose was identified with the modern CGM and was found to deviate substantially from the new suggested formula. More importantly, we found a more robust correlation between HbA1c and CGM levels in patients with type 2 diabetes. Overestimation or underestimation of blood glucose levels through CGM might increase the risks of inappropriate clinical treatment of diabetes patients. Our results indicate the need for proper CGM data interpretation individualized for each patient to better assist the determination of customized treatments for patients.

Background Although generally considered part of a healthy diet, coffee consumption has been suspected to be associated with elevated epinephrine levels and increasing insulin resistance. Objectives We studied the effects of the intake of 3 different types of coffee (Tanzanian, Ethiopian, and Kenyan) on postprandial interstitial glucose levels. Method Interstitial glucose levels were measured every 15 minutes using the FreeStyle Libre glucose monitoring system (Abbott Diabetes Care Ltd, Witney, United Kingdom) in each individual after drinking coffee compared with when not consuming coffee. Results Unlike Tanzanian and Ethiopian coffees, Kenyan coffee suppressed the increase of postprandial interstitial glucose levels. Kenyan coffee beans contain less anhydrous caffeine and more chlorogenic acid than Tanzanian and Ethiopian coffee beans. These findings may explain the different effects of these coffee types on postprandial interstitial glucose levels. Furthermore, Kenyan coffee beans inhibited α-glucosidase activity, which may partially explain why Kenyan coffee reduces postprandial interstitial glucose levels. Conclusions Coffee is widely consumed as a beverage worldwide, and our findings suggest that patients with diabetes mellitus may benefit from drinking Kenyan coffee because of its ability to reduce postprandial interstitial glucose levels. (Curr Ther Res Clin Exp. 2020; 81:XXX–XXX)

COVID-19 pandemic poses problems that not only concern the economy but also the health of people all over the world. In Japan, despite the declaration of a “state of emergency”, no lockdown was implemented, and a request for self-restraint and avoidance of non-essential trips was instead issued. After a month, the state of emergency was lifted. Because patients with diabetes mellitus (DM) were forced to stay during the state of emergency, resulting in a lack of physical activity, concerns about their glycemic control were raised. Therefore, glycated hemoglobin (HbA1c) levels during different time periods were compared (May 2018, March 2019, June 2019, July 2019, May 2019, March 2020, June 2020, July 2020). We analyzed 165 patients with DM. The mean age of subjects was 67.8 + 11.5 years. Male comprised 67.3% of the participants. The mean body weight was 65.6 + 14.6 kg on July 2019 and 66.1 + 15.2 kg on July 2020. The mean body mass index (BMI) was 24.4 + 3.6 kg/m2 on July 2019 and 24.4 + 3.6 on July 2020. Patients with Type 2 DM (T2DM) comprised 90% of the participants, while the rest had T1DM. Mean duration of DM was 12.0 + 7.4 years. In order to assess the effect of the self-restraint on plasma glucose control, HbA1c levels during these periods were compared: May 2018, March 2019, June 2019, July 2019 (one year before COVID-19 pandemic.), and May 2019, March 2020, June 2020, July 2020 (The last three months during COVID-19). March 2020 is corresponded to a period before the request for self-restraint, while June and July 2020 corresponded to the periods right after the end of self-restraint. We also compared HbA1c levels between May 2019 and July 2020 using the Self-Monitoring of Blood Glucose (SMBG) to assess whether SMBG affected plasma glucose control during the period of self-restraint. HbA1c levels in May 2018, March 2019, June 2019, July 2019, May 2019, March 2020, June 2020, July 2020, were 7.32 + 1.23, 7.44 + 1.20, 7.16 + 1.06, 7.01 + 1.05, 7.23 + 1.06, 7.45 + 1.18, 7.15 + 10.7, and 7.11 + 1.17, respectively. Similarly, HbA1c levels between May 2019 without SMBG and May 2019 with SMBG were not statistically different. In this clinical study, we found that the request to avoid non-essential trips as a form of self-restraint during the country’s state of emergency did not affect plasma glucose control of patients with DM. We noted that the patients did not have signs of insulin resistance as their BMI on July 2019 and July 2020 were 24.4 + 3.6 and 24.4 + 3.6, respectively. Unexpectedly, the HbA1c levels were not affected by the absence or presence of SMBG. This could explain why HbA1c levels were not elevated, despite a temporarily sedentary lifestyle and a lack of exercise for a month. In addition, due to the self-restraint, the frequency of dining outside the house decreased, which could have contributed to the non-elevation of HbA1c levels.

Nucleobindin-2 (NUCB2) is a 396-amino acid protein, cleaved into the N-terminal nesfatin-11-82, nesfatin-285-163 and the C-terminal nesfatin-3166-396. NUCB2 contains a signal peptide, a leucine zipper structure, two Ca2+ binding EF-hand domains, and has a wide variety of basic cellular functions. NUCB2 is also a precursor protein of nesfatin-1, which was originally identified in hypothalamic nuclei, and which is a regulatory factor involved in the central control of food intake and energy balance. There are several reports indicating that NUCB2 is also expressed in various human peripheral tissues. Moreover, recent studies have reported that high levels of NUCB2 mRNA and protein are a potent prognostic factor for prostate cancer, endometrial carcinoma, and breast cancer. NUCB2 was also identified as a potential tumor antigen eliciting autoantibody responses in 5.4% of gastric cancer patients but not in the healthy individuals. However, theclinicopathological significance of NUCB2 expression in gastric cancer has still not been elucidated. Therefore, we examined NUCB2 expression in a large number of gastric cancer patients, using immunohistochemistry, to explore its clinicopathological significance. To explore this, we aimed to investigate the NUCB2 expression in gastric cancer tissues and adjacent non-tumor tissues and its potential relevance to clinicopathological factors and prognosis using immunohistochemistry analysis. In our study, NUCB2 level in gastric cancer tissues was higher than in non-tumor tissues. A high expression of NUCB2 is significantly associated with tumor depth, lymph node metastasis, lymphatic invasion, venous invasion and clinical stage. Furthermore, the expression level of NUCB2 protein was independent predictor of progression-free survival. In summary, NUCB2 might play a crucial role in gastric cancer development and could serve as an independent predictor of prognosis of gastric cancer patients.

Background: Establishing an optimal insulin regimen is crucial for maintaining glycemic control in patients with type 1 diabetes (T1D). The aim of the present study was to determine the insulin dose required to achieve an HbA1c concentration ≤7.5% in Japanese patients with T1D. Methods: The present multicenter cross-sectional study was performed at three institutes in Japan. Information was collected regarding patient age, sex, body weight, body mass index (BMI), HbA1c, total daily insulin dose (TDD), and total basal insulin dose (TBD), and the effects of these factors on achieving HbA1c ≤7.5% were investigated. Results: Of 107 patients with T1D, 92 had no detectable endogenous insulin secretion: 39 had HbA1c ≤7.5% (well-controlled group) and 53 had HbA1c >7.5% (poorly controlled group). No significant differences in age, sex, height, body weight, BMI, diabetes duration, stage of diabetic kidney disease, treatment, or TDD were noted between the poorly and well-controlled groups. The TBD as a percentage of TDD (%TBD) was lower in patients with well-controlled diabetes ( P < 0.05) after adjustment for age, gender, and diabetes duration. In the well-controlled group, TDD was correlated with body weight ( R = 0.51), BMI ( R = 0.44), body surface area ( R = 0.41), and TBD ( R = 0.73; P < 0.01 for all), but TBD was not correlated with BMI or body surface area. In our population, a %TBD of approximately 30% was appropriate, without considering BMI. Conclusions: To achieve HbA1c ≤7.5 in patients with T1D, TDD should be calculated based on body weight, and the %TBD should be set at 30% in the Japanese population.

Somatic mutations in KCNJ5 gene have been identified in patients with adrenal aldosterone-producing adenomas (APAs). We previously reported that Japanese patients with APAs had distinct characteristics from patients in Western countries; i.e. they had a high frequency of KCNJ5 mutations and exhibited a frequent association with cortisol co-secretion. Therefore, APAs among Japanese patients may have different features from those in Western countries. We added recent cases, examined 47 cases (43% male) of APAs, including clinicopathological features, KCNJ5 mutations, and the mRNA levels of several steroidogenic enzymes, and compared the results obtained to those reported in other countries. While the prevalence of KCNJ5 mutations is approximately 40% in Western countries, 37 APA cases (78.7%) showed mutations: 26 with p.G151R and 11 with p.L168R. Although a significant gender difference has been reported in the frequency of KCNJ5 mutations in Europe, we did not find any gender difference. However, the phenotypes of Japanese patients with mutations were similar to those of patients in Western countries; patients were younger and had higher plasma aldosterone levels, lower potassium levels, and higher diastolic blood pressure. Reflecting these phenotypes, APAs with mutations had higher CYP11B2 mRNA levels. However, in contrast to APAs in Western countries, Japanese APAs with mutations showed lower CYP11B1, CYP17A1, and CYP11A1 mRNA levels. These findings demonstrated that Japanese APA patients may have distinct features including a higher prevalence of KCNJ5 mutations, no gender difference in the frequency of these mutations, and characteristics similar to the zona glomerulosa.

Aims Optimum therapy for patients with diabetes depends on both acute and long-term changes in plasma glucose, generally assessed by HbA1c levels. However, the correlation between HbA1c and circulating glucose has not been fully determined. Therefore, we carefully examined this correlation when glucose levels were assessed by continuous glucose monitoring (CGM). Methods 51 patients (70 % women, 30% male) were examined; among them were 28 type 1 patients with diabetes and 23 type 2 patients with diabetes. Clinically determined HbA1c levels were compared with blood glucose determined by CGM during a short time period. Results: Changes of HbA1c levels up to 8.0% showed a clear and statistically strong correlation (R=0.6713, P Results Changes of HbA1c levels up to 8.0% showed a clear and statistically strong correlation (R=0.6713, P Conclusions Short term CGM appears to be a good clinical indicator of long-term glucose control (HbA1c levels), however cautions should be taken while interpreting CGM data from patients with HbA1c levels above 8.0%. Over or under estimation of the actual mean glucose from CGM data could potentially increase the risks of inappropriate treatment. As such, our results indicate that a more accurate analysis of CGM data might be useful to adequately tailor clinical treatments.

Expression of adaptor protein, phosphotyrosine interaction, pleckstrin homology domain, and leucine zipper containing 1 (APPL1) promoted glucose transporter 4 (GLUT4) translocation and glucose uptake in adipose and muscle tissues in response to stimulation with insulin, adiponectin, or exercise. In response to mechanical stretch, knockdown of APPL1 in C2C12 myotubes suppressed glucose uptake. APPL1-induced increased glucose uptake was mediated by protein kinase C (PKC) ζ but not AKT, AMPK, or calmodulin-dependent protein kinase. In myotubes overexpressing APPL1, PKCζ was phosphorylated and translocated to the plasma membrane (PM) in response to mechanical stretch. Phosphorylated PKCζ co-immunoprecipitated with protein phosphatase 2A (PP2A) under basal conditions, but dissociated upon myotube stretching. Moreover, stretch-induced phosphorylated PKCζ co-immunoprecipitated with non-muscle myosin IIa. Blebbistatin, an inhibitor of myosin II ATPase activity, suppressed APPL1-mediated stretch-induced glucose uptake and PKCζ translocation. Taken together these data demonstrate that in response to mechanical stretch, APPL1 enhances glucose uptake by modulating the activation and localization of PKCζ, as well as its functional interaction with both PP2A and myosin IIa. These findings support a new function for non-muscle myosin IIa in differentiated myotubes.

Sodium glucose co-transporter-2 (SGLT2) inhibitors are used to reduce glycated hemoglobin (HbA1c) levels. Although they have the potential to reduce body weight in patients with type 2 diabetes mellitus (T2DM), some patients may gain weight. We found that the effects of the SGLT2 inhibitor ipragliflozin were maintained for 48 h when used in combination with nifedipine, so we considered that a daily SGLT2 inhibitor might be unnecessary. Therefore, we investigated whether as-required single doses of an SGLT2 inhibitor would be clinically relevant in three patients having difficulty maintaining blood glucose control when they were unable to exercise or during social situations. All three patients demonstrated improved HbA1c levels with as-required single doses of an SGLT2 inhibitor. These patients experienced no adverse effects or weight gain relative to taking an SGLT2 inhibitor daily. Thus, as-required single-dose SGLT2 inhibitors could be one of the many options to improve blood glucose control in T2DM.

FAM19A2/TAFA-2, a member of the chemokine CC family, shares 31% sequence identity with MIP-1α, which is known to elevate body temperature and reduce food intake. A single administration of 250 pM of FAM19A2/TAFA-2 to the third ventricle of mice just before the initiation of dark period increased food intake and meal number significantly, but reduced meal size during the dark period. The respiratory exchange rate and energy expenditure were increased significantly during the dark period, while the ambulatory count and vertical activity were not affected. These data suggest that FAM19A2/TAFA-2 participates in the regulation of food intake and metabolic activities.

We reported a positive correlation between brachial-ankle pulse wave velocity (baPWV) and one-hour post-challenge glucose (1-h PG). Furthermore, the subpopulation defined by 1-hour PG beyond 183 mg/dL (corresponded to Impaired glucose tolerance range of 2-h PG) in normal glucose tolerant (NGT) subjects. These findings suggested that if NGT subjects convert to higher 1-h PG levels from a normal 1-h PG range they will develop arterial stiffness. To address this issue, we initiated a clinical study that included 40 NGT subjects (23 men and 17 women, mean age 60.9 ±9.7 years), with a comprehensive medical check-up was performed on each subject at Kan-etsu Chuo Hospital every year. In addition, over the 9-year period these individuals were annually assessed by a 75-g oral glucose tolerance test (OGTT) and for atrial stiffness by baPWV. Based on the 75-g OGTT, NGT subjects alone were selected according to the 2006 WHO criteria. NGT subjects in this study were identified as individuals having a normal range of all measured parameters. In this population, there was no significant correlation between baPVW and all measured parameters. One-h post challenge glucose (1-h PG) in 28 subjects was below 154 mg/dL and their baPWV was not changed (1266.4 ±125.42 cm/sec from 1243.62 ±108.1) after nine years. On the other hand, 1-h PG in the rest of 12 subjects was significantly increased to 160.33 ±4.73 from 124.67 ±20.13 and their baPVW was also significantly elevated to 1489.33 ±41.05 from 1255.01 ±64.08 after nine years. We found that rapid blood glucose change in NGT could be a risk factor for arterial stiffness and we would like to point out the possibility for physicians to select high-risk subjects by 75g-OGTT with the evaluation of 1h-PG. Taken together, we found that rapid blood glucose change could be a risk factor for arterial stiffness. Disclosure S. Okada: None. J. Okada: None. E. Yamada: None. T. Saito: None. M. Yamada: None.

Background: The Abbott FreeStyle Libre Flash glucose monitoring system (FGM) is one of the recently introduced personal continuous glucose monitoring systems. While FGM use has spread widely, few reports have examined the factors that determine its accuracy. Methods: We performed oral glucose tolerance tests (OGTTs) in 25 Japanese subjects without diabetes. Results: The mean absolute relative difference (MARD) in sensor glucose values of FGM (FGM-SG) with reference to self-monitoring of blood glucose (SMBG) levels was 10.9±4.1% during OGTTs. However, surprisingly, the MARD in FGM-SG with reference to plasma glucose (PG) levels was 20.3±10.3% during OGTTs, demonstrating the problems in accuracy of FGM-SG compared with that of PG levels in our study setting; however, this issue was not observed on comparison with accuracy of SMBG levels. Parkes error grid analyses showed that FGM-SG with either SMBG or PG levels as reference met the criteria of the International Organization for Standardization. Moreover, the MARD showed negative correlations with fasting blood sugar, homeostasis model assessment insulin resistance index, area under curve (AUC)-PG, and body mass index (BMI); multiple regression analyses revealed that BMI was the most contributing factor to the MARD between FGM-SG and PG levels. In other words, lean people showed more MARDs. Conclusions: Inaccurate FGM data could potentially increase the risk of inappropriate treatment. Consideration of such factors might be critical while using FGM. Disclosure S. Yoshino: None. E. Yamada: None. S. Okada: None. T. Saito: None. M. Yamada: None.

We have reported that a patient with painless thyroiditis induced by the cessation of dipeptidyl peptidase-4 inhibitor was accompanied with type 2 diabetes mellitus (T2DM). Given the lack of clarity regarding the possibility whether dipeptidyl peptidase-4 inhibitors (DPP-4is) attenuates thyroid hormone levels, we compared thyroid-stimulating hormone (TSH) and free thyroxine (FT4) levels between patients with T2DM who were not prescribed DPP-4is and patients with T2DM who were prescribed DPP-4is. 12 patients were prescribed DPP-4is (DPP-4is group) and 33 patients were not prescribed DPP-4is (w/o DPP-4is group). TSH and FT4 levels were measured in all of these patients and TSH and FT4 levels were annually measured in all of patients in the current population for more than three years. The TSH levels in the DPP-4is group was significantly higher than those of w/o DPP-4is group (2.11 ±0.67 vs1.58 ±0.64 μIU/mL, mean ±standard deviation, reference range: 0.33~4.05, p= 0.0189). The FT4 levels in the DPP-4is group was not significantly different from those of w/o DPP-4is group (1.06 ±0.086 vs1.01 ±0.12 ng/dL, mean ±standard deviation, reference range: 0.97~1.69, p= 0.0189). The measured TSH and FT4 levels were normal range in all of subjects in this clinical study. Nevertheless, TSH levels in patients with T2DM prescribed DPP-4is was significantly higher than those of the patients with T2DM not prescribed DPP-4is although we did not see the difference in FT4 levels between two groups. Despite of the small sample size, this clinical study provides important information. One strength of the current study is that the TSH and FT4 levels were annually measured in all patients with T2DM in the current population for more than three years. DPP-4is might modulate TSH levels in the absence of Metformin and further investigates with large population were necessary whether we need to monitor thyroid function regularly when we prescribe DPP-4is for the patients with T2DM. Disclosure Y. Shimoda: None. J. Okada: None. E. Yamada: None. T. Saito: None. S. Okada: None. M. Yamada: None.

Context We previously identified factors affecting thyroid status, including sex, age, and smoking. Objective In the current study, we increased the number of subjects examined and investigated the effects of these factors, particularly smoking and the thyroid peroxidase antibody (TPO-Ab), in Japanese patients with euthyroxinemia and serum free T4 levels within the normal range. Participants A total of 12,289 subjects who underwent health checkups were analyzed in a cross-sectional and longitudinal study. Results The mean age of subjects was 50 ± 10 years (age range: 21 to 88 years). Serum TSH levels and the prevalence of positivity for TPO-Ab increased with age in Japanese subjects with euthyroxinemia. Mean serum TSH levels were significantly lower in the smoking group than in the nonsmoking group except for women older than 50 years. Serum TSH levels were significantly higher in subjects with positivity for TPO-Ab than in those with negativity at all ages and in both sexes; however, smoking did not affect free T4 levels or positivity for TPO-Ab. Among men, the rate of smokers was significantly higher in patients with subclinical hyperthyroidism (25%) than in those with subclinical hypothyroidism (10%; P < 0.05). Furthermore, the results of the longitudinal study revealed a significant decrease in serum TSH levels 1 year after the start of smoking in men (P < 0.05). Conclusion Because smoking appeared to lower serum TSH levels in Japanese subjects with euthyroxinemia, their smoking status warrants careful consideration when evaluating subclinical thyroid function.

Aim: Thyroid hormone affects the whole-body metabolism, including that of lipids and glucose, and its involvement in NAFLD / NASH has recently been reported. Accurate evaluation of thyroid function is therefore important. In this study, seasonal changes in thyroid function in approximately 10,000 health check participants were investigated, and the effects of age and smoking were also examined.Subjects and methods: The subjects were participants of medical checkups at Hidaka Hospital from 2007 to 2013. Exclusion criteria were a past history of thyroid disease, steroid use, renal failure and liver cirrhosis. To exclude the effects of menstruation, we examined 8,489 men whose blood TSH and FT4 values were within the reference values for monthly changes in thyroid function, and the effects of age and smoking. All blood TSH and FT4 levels were measured at 8 ~ 9:00 AM after over night fasting.Results: The average age was 49 ± 10 years old. The number of participants varied each month, with a minimum of 346 in April and a maximum of 961 in November. The highest median serum TSH level was 1.6 μU/ml in January (winter), and the lowest was 1.3 μU/ml in June, July, August and September (summer). These values were significantly different from the yearly median value of 1.4 μU/ml in April and November. On the other hand, the FT4 level was significantly lower in August than in the other months except for February. The extents of seasonal changes in TSH and FT4 levels were reduced in elderly subjects >60 years old. Furthermore, the median blood TSH level of 2,262 smokers was lower than that of 4,709 non-smokers (median: smoker, 1.2μU/ml vs. non-smoker, 1.5μU/ml). However, seasonal changes were also observed in smokers. Conclusions: This is the first report in a large healthy cohort demonstrating that there are significant seasonal changes in thyroid function including freeT4 levels. Several factors, such as season, age, smoking and gender, need to be taken into account for accurate evaluation of thyroid function.

Brief refeeding times (~60 min) enhanced hepatic Angptl8 expression in fasted mice. We cloned the mouse Angptl8 promoter region to characterise this rapid refeeding-induced increase in hepatic Angptl8 expression. Deletion of the −309/−60 promoter region significantly attenuated basal promoter activity in hepatocytes. A computational motif search revealed a potential binding motif for hepatocyte nuclear factor 1α/1β (HNF-1α/β) at −84/−68 bp of the promoter. Mutation of the HNF-1 binding site significantly decreased the promoter activity in hepatocytes, and the promoter carrying the mutated HNF-1 site was not transactivated by co-transfection of HNF-1 in a non-hepatic cell line. Silencing Hnf-1 in hepatoma cells and mouse primary hepatocytes reduced Angptl8 protein levels. Electrophoretic mobility-shift assays confirmed direct binding of Hnf-1 to its Angptl8 promoter binding motif. Hnf-1α expression levels increased after short-term refeeding, paralleling the enhanced in vivo expression of the Angptl8 protein. Chromatin immunoprecipitation (ChIP) confirmed the recruitment of endogenous Hnf-1 to the Angptl8 promoter region. Insulin-treated primary hepatocytes showed increased expression of Angptl8 protein, but knockdown of Hnf-1 completely abolished this enhancement. HNF-1 appears to play essential roles in the rapid refeeding-induced increases in Angptl8 expression. HNF-1α may therefore represent a primary medical target for ANGPTL8-related metabolic abnormalities. The study revealed the transcriptional regulation of the mouse hepatic Angptl8 gene by HNF-1.

In the present study we examined the efficacy of sodium‐glucose cotransporter 2 inhibitors on improvement of glycated hemoglobin (HbA1c) in comparison with the renal threshold for glucose reabsorption in patients with type 2 diabetes mellitus. Patients visited the hospital once a month for a regular follow‐up examination with the determination of blood glucose and HbA1c levels, and urinary glucose concentration from spot urine samples. Patient samples were compared before and after ipragliflozin administration. We defined the renal threshold for glucose reabsorption as the lowest blood glucose level that correlated with the first detectable appearance of urine glucose. These data showed a significant negative correlation between improvement of HbA1c level and renal threshold for glucose reabsorption in patients treated with the sodium‐glucose cotransporter 2 inhibitor. These findings show that patients who have a higher renal threshold for glucose reabsorption can be expected to more effectively respond to sodium‐glucose cotransporter 2 inhibitor therapy in terms of lowering HbA1c levels.