Vgll2 as an integrative regulator of mitochondrial function and contractility specific to skeletal muscle.

During skeletal muscle adaptation to physiological or pathophysiological signals, contractile apparatus and mitochondrial function are coordinated to alter muscle fiber type. Although recent studies have identified various factors involved in modifying contractile proteins and mitochondrial function, the molecular mechanisms coordinating contractile and metabolic functions during muscle fiber transition are not fully understood. Using a gene-deficient mouse approach, our previous studies uncovered that vestigial-like family member 2 (Vgll2), a skeletal muscle-specific transcription cofactor activated by exercise, is essential for fast-to-slow adaptation of skeletal muscle. The current study provides evidence that Vgll2 plays a role in increasing muscle mitochondrial mass and oxidative capacity. Transgenic Vgll2 overexpression in mice altered muscle fiber composition toward the slow type and enhanced exercise endurance, which contradicted the outcomes observed with Vgll2 deficiency. Vgll2 expression was positively correlated with the expression of genes related to mitochondrial function in skeletal muscle, mitochondrial DNA content, and protein abundance of oxidative phosphorylation complexes. Additionally, Vgll2 overexpression significantly increased the maximal respiration of isolated muscle fibers and enhanced the suppressive effects of endurance training on weight gain. Notably, no additional alteration in expression of myosin heavy chain genes was observed after exercise, suggesting that Vgll2 plays a direct role in regulating mitochondrial function, independent of its effect on contractile components. The observed increase in exercise endurance and metabolic efficiency may be attributed to the acute upregulation of genes promoting fatty acid utilization as a direct consequence of Vgll2 activation facilitated by endurance exercise. Thus, the current study establishes that Vgll2 is an integrative regulator of mitochondrial function and contractility in skeletal muscle.

Transcriptome and secretome profiling of sensory neurons reveals sex differences in pathways relevant to insulin sensing and insulin secretion.

Sensory neurons in the dorsal root ganglia (DRG) convey somatosensory and metabolic cues to the central nervous system and release substances from stimulated terminal endings in peripheral organs. Sex-biased variations driven by the sex chromosome complement (XX and XY) have been implicated in the sensory-islet crosstalk. However, the molecular underpinnings of these male-female differences are not known. Here, we aim to characterize the molecular repertoire and the secretome profile of the lower thoracic spinal sensory neurons and to identify molecules with sex-biased insulin sensing- and/or insulin secretion-modulating activity that are encoded independently of circulating gonadal sex hormones. We used transcriptomics and proteomics to uncover differentially expressed genes and secreted molecules in lower thoracic T5-12 DRG sensory neurons derived from sexually immature 3-week-old male and female C57BL/6J mice. Comparative transcriptome and proteome analyses revealed differential gene expression and protein secretion in DRG neurons in males and females. The transcriptome analysis identified, among others, higher insulin signaling/sensing capabilities in female DRG neurons; secretome screening uncovered several sex-specific candidate molecules with potential regulatory functions in pancreatic ß cells. Together, these data suggest a putative role of sensory interoception of insulin in the DRG-islet crosstalk with implications in sensory feedback loops in the regulation of ß-cell activity in a sex-biased manner. Finally, we provide a valuable resource of molecular and secretory targets that can be leveraged for understanding insulin interoception and insulin secretion and inform the development of novel studies/approaches to fathom the role of the sensory-islet axis in the regulation of energy balance in males and females.

Assembled Structures and Electrical Conductivities of Salts Comprised of Cationic Sandwich Complexes and Polyoxometalates.

Polyoxometalates (POMs) are recognized for their diverse structures and catalytic properties, yet their organometallic salts have not been thoroughly investigated. In this study, we synthesized salts of a Keggin-type POM with cationic sandwich complexes featuring various substituents, [X]2[SMo12O40] [X = Ru(C5H5)(C6H5R); R = H (1a), Me (1b), Bu (1c)], [Co(C5H5)2 (2), and Co(C5Me5)2 (3)]. These salts exhibited similar structural characteristics, with each anion surrounded by 10 cations, forming two-dimensional sheet arrangements through O···O anion-anion contacts, except for the salt of 1c, which exhibited one-dimensional contact owing to the larger cation substituent. Additionally, [2](THA)[Mo6O19], [3]2[Mo6O19], and [3](THA)[SMo12O40], containing the Lindqvist-type POM [Mo6O19] and/or tetrahexylammonium (THA) cation, were obtained, with the packing structure of [3]2[Mo6O19] closely resembling that of [X]2[SMo12O40]. Solvate crystals of [1a]3[PMo12O40] were also prepared. [1a]2[SMo12O40] exhibits an electrical conductivity σ of 1.4 × 10-7 S cm-1 at 373 K, which is 2 orders of magnitude higher than those of (THA)2[SMo12O40] and [1a]PF6.

Organometallic Ionic Plastic Crystals Incorporating Cationic Half-Sandwich Complexes.

Ionic plastic crystals (IPCs), characterized by nearly spherical molecular ions, exhibit remarkable solid-state characteristics including high ionic conductivity. However, most IPCs are organic onium salts. Incorporating organometallic half-sandwich complexes into IPCs is challenging owing to their low-symmetry structures. This paper introduces a novel series of IPCs composed of salts derived from half-sandwich organometallic complexes. We synthesized five salts of [Ru(Cp)(tmeda)(CO)]X (tmeda = N,N,N',N'-tetramethyl-1,2-ethanediamine, X = anion) with different anions and examined their phase behavior, crystal structures, and molecular motion in the solid-state. Salts featuring the CPFSA (= 1,1,2,2,3,3-hexafluoropropane-1,3-disulfonimide), B(CN)4-, and FSA- (= (FSO2)2N-) anions underwent phase transitions to an IPC phase with a CsCl-type structure in the temperature range of 327-364 K. Employing smaller anions led to an increase in the transition temperature. In each salt, the coordination number, representing the number of anions surrounding one cation, remained eight in IPC and low-temperature phases. However, salts containing smaller anions (CF3BF3- and PF6-) displayed a rotator phase rather than the IPC phase. In these cases, the coordination numbers were six at low temperatures.

What Happens 20 to 30 years After Radial Keratotomy? Case Series.

ABSTRACT: Radial keratotomy (RK) was commonly performed in the 1980s and 1990s. We aimed to clarify the current status of post-RK refractive correction and treatment. We retrospectively reviewed the charts of 70 patients with a history of RK. Of the 70 patients, 44 were identified for clinical outcomes. Refractive or therapeutic intervention (rigid gas-permeable contact lens fit, spectacle prescription, corneal surgery, and use of pilocarpine hydrochloride for photophobia) was possible in 59% of patients with postoperative visual deterioration after RK; in the remaining 41%, therapeutic intervention was not possible. Rigid gas-permeable contact lens fit for corneal irregular astigmatism was the most common refractive intervention and was effective in 36% of cases in the university hospital.

A Comprehensive Wavefront Assessment of Keratoconus Using an Integrated Scheimpflug Corneal Tomographer/Hartmann-Shack Wavefront Aberrometer.

OBJECTIVES: To characterize higher-order aberrations (HOAs) in different severities of keratoconus (KC) from the anterior and posterior corneal surfaces and whole eye using an integrated Scheimpflug corneal tomographer/Hartmann-Shack wavefront aberrometer. METHODS: This study included eyes with clinical KC, topographic KC (no clinical signs), fellow eyes with very asymmetric ectasia with normal topography and no clinical signs (VAE-NT), and control eyes. Corneal and ocular wavefront aberrations were obtained using an integrated Scheimpflug tomographer/Hartmann-Shack wavefront aberrometer. The diagnostic capability of distinguishing VAE-NT from the control was also tested. RESULTS: This study included 68 eyes with clinical KC, 44 with topographic KC, 26 with VAE-NT, and 45 controls. Clinical KC had significantly greater total HOAs and coma from the anterior and posterior corneal surfaces and whole eye than the other groups ( P <0.05). Although topographic KC had significantly greater values in all wavefront parameters than the control ( P <0.05), ocular and corneal HOAs did not differ between the VAE-NT and control groups. The coma from the anterior cornea in topographic KC was significantly greater than that in VAE-NT ( P <0.05); the coma from the posterior cornea and whole eye did not differ. Total HOAs from the anterior corneal surface exhibited the highest area under the receiver operating characteristic curve value of 0.774 (sensitivity, 73%; specificity, 78%). CONCLUSION: A comprehensive wavefront assessment can be used to quantitatively evaluate corneal and ocular HOAs across various severity of KC. Total HOAs from the anterior corneal surface exhibited the potential ability in distinguishing VAE-NT from the control eyes.

Comparison of Ocular Wavefront Aberration Measurements Obtained Using Two Hartmann-Shack Wavefront Aberrometers.

OBJECTIVES: To assess agreement between measurements of ocular wavefront aberrations obtained using the Pentacam AXL Wave (Oculus Optikgeräte GmbH) (Aberrometer A) and KR-1W (Topcon Corp) (Aberrometer B), both of which are based on the Hartmann-Shack principle. METHODS: In this prospective case-control study, ocular wavefront aberrations measurements were obtained using both aberrometers in patients with keratoconus (KC) and control participants. Ocular wavefront aberrations were measured through the natural pupil without dilation using both devices in a dark room. For both aberrometers, accommodation was inhibited by automatically adding fogging. The individual Zernike coefficients from the second to fourth order were compared between the two aberrometers for a 4-mm pupil diameter. RESULTS: Twenty-six KC and 29 control eyes were assessed. Statistically significant correlations ( P <0.05) were observed for all Zernike coefficients, except for Z 4-2 in the control group. Bland-Altman analysis indicated good agreement between aberrometers and no statistically significant differences in the control group. However, in the KC group, patterns of proportional error were observed in vertical coma Z 3-1 (r=0.338, P =0.008), trefoil Z 4-4 (r=0.701, P =0.003), secondary astigmatism Z 4-2 (r=0.348, P =0.025), and spherical aberrations Z 40 (r=0.407, P =0.012). CONCLUSIONS: The Zernike coefficient values measured by the two aberrometers were well correlated in the control and KC groups. However, in eyes with KC, Aberrometer B tended to present greater values in several Zernike coefficients than Aberrometer A, suggesting that wavefront measurements obtained using the two aberrometers are not interchangeable in patients with KC.

Long-Term Corneal Refractive Power Changes Two Decades After Radial Keratotomy With Microperforations.

ABSTRACT: We retrospectively examined corneal refractive power in three patients who had been followed up for more than 20 years after radial keratotomy (RK) with microperforations (MPs). All patients underwent RK in both eyes and were referred to our clinic because of postoperative decreased vision. MP was observed in five of the six eyes at the initial visit. The corneal refractive power of the anterior and posterior surfaces of the 6-mm-diameter cornea was examined using Fourier analysis based on corneal shape analysis using anterior segment optical coherence tomography. The spherical components decreased in all three cases. The asymmetry and higher-order irregularity components and fluctuations in corneal refractive power were markedly greater in the two cases with MP in both eyes. Fluctuations in corneal refractive power were observed at more than 20 years after RK with MP. Therefore, careful observation is necessary, even after a long-term postoperative follow-up period.

The multifaceted role of ATF4 in regulating glucose-stimulated insulin secretion.

Stress-inducible transcription factor ATF4 is essential for survival and identity of ß-cell during stress conditions. However, the physiological role of ATF4 in ß-cell function is not yet completely understood. To understand the role of ATF4 in glucose-stimulated insulin secretion (GSIS), ß-cell-specific Atf4 knockout (ßAtf4KO) mice were phenotypically characterized. Insulin secretion and mechanistic analyses were performed using islets from control Atf4f/f and ßAtf4KO mice to assess key regulators for triggering and amplifying signals for GSIS. ßAtf4KO mice displayed glucose intolerance due to reduced insulin secretion. Moreover, ßAtf4KO islets exhibited a decrease in both the insulin content and first-phase insulin secretion. The analysis of ßAtf4KO islets showed that ATF4 is required for insulin production and glucose-stimulated ATP and cAMP production. The results demonstrate that ATF4 contributes to the multifaceted regulatory process in GSIS even under stress-free conditions.

Characteristics of Higher-Order Aberrations in Different Stages of Keratoconus.

OBJECTIVES: To characterize higher-order aberrations (HOAs) in clinical and subclinical keratoconus (KC). METHODS: The study included 33, 36, and 26 patients with clinical, topographic (no clinical signs), and pretopographic (normal topography and no clinical signs) KC and 30 controls. Ocular and corneal HOAs for the 4-mm pupils were measured using a wavefront sensor and expanded up to the sixth order of Zernike polynomials. The magnitudes of trefoil, coma, tetrafoil, secondary astigmatism, and spherical aberration were calculated via Zernike vector analysis and used as HOA parameters along with total HOAs. Area under the receiver operating characteristic curve (AUROC) values for each wavefront parameter for pretopographic KC were compared. RESULTS: Control eyes and eyes with pretopographic KC had significantly lower ocular or corneal total HOAs and Zernike vector terms than those with clinical KC and topographic KC, except for ocular tetrafoil between topographic KC and pretopographic KC and spherical aberration among all groups. The AUROCs for corneal total HOAs and corneal coma for pretopographic KC and control eyes were 0.781 (100% sensitivity and 47% specificity) and 0.735 (73% sensitivity and 73% specificity), respectively. CONCLUSION: Corneal total HOAs and corneal coma exhibited a potential ability to discriminate pretopographic KC from normal control eyes.

The Characteristics and Risk Factors of Very Asymmetric Keratoconus.

PURPOSE: To characterize very asymmetric keratoconus (KC) in terms of clinical demographics and risk factors in the Japanese population. METHODS: We retrospectively reviewed the clinical records of patients with very asymmetric KC attending a university hospital. Patients with very asymmetric KC had defined clinical KC in one eye and normal topography in the fellow eye. All patients completed a questionnaire on potential risk factors (family history of KC, atopy, asthma, allergy, heart disease, sleep apnea, Down syndrome, eye rubbing, prone sleep position, and dominant hand). These data were compared with those of patients with clinical or topographic KC in both eyes. Subgroup analysis of very asymmetric KC eyes were performed based on the KC status: mild and severe. RESULTS: We retrospectively investigated 66 patients with very asymmetric KC and 505 patients with KC. Patients reported eye rubbing (53%-56%), allergy (62%-63%), and atopy (28%-29%) in both groups. There was no significant difference in terms of risk factors between the two groups. Approximately half of the 66 patients with very asymmetric KC had KC eyes ipsilateral to the dominant hand. Subgroup analysis of very asymmetric KC showed that atopy and asthma were more frequent in the mild KC group than in the severe KC group. Forty-four of 60 patients (73.3%) were using corneal, rigid, gas-permeable contact lenses (corneal GPs) for the KC eye, of which 30 patients (68.2%) were also using a corneal GP for the fellow eye. CONCLUSIONS: The patient-reported frequency of eye rubbing, allergy, and atopy were similar between KC and very asymmetric KC. Furthermore, we found no association between hand dominance and KC laterality.

Linagliptin Ameliorates Hepatic Steatosis via Non-Canonical Mechanisms in Mice Treated with a Dual Inhibitor of Insulin Receptor and IGF-1 Receptor.

Abnormal hepatic insulin signaling is a cause or consequence of hepatic steatosis. DPP-4 inhibitors might be protective against fatty liver. We previously reported that the systemic inhibition of insulin receptor (IR) and IGF-1 receptor (IGF1R) by the administration of OSI-906 (linsitinib), a dual IR/IGF1R inhibitor, induced glucose intolerance, hepatic steatosis, and lipoatrophy in mice. In the present study, we investigated the effects of a DPP-4 inhibitor, linagliptin, on hepatic steatosis in OSI-906-treated mice. Unlike high-fat diet-induced hepatic steatosis, OSI-906-induced hepatic steatosis is not characterized by elevations in inflammatory responses or oxidative stress levels. Linagliptin improved OSI-906-induced hepatic steatosis via an insulin-signaling-independent pathway, without altering glucose levels, free fatty acid levels, gluconeogenic gene expressions in the liver, or visceral fat atrophy. Hepatic quantitative proteomic and phosphoproteomic analyses revealed that perilipin-2 (PLIN2), major urinary protein 20 (MUP20), cytochrome P450 2b10 (CYP2B10), and nicotinamide N-methyltransferase (NNMT) are possibly involved in the process of the amelioration of hepatic steatosis by linagliptin. Thus, linagliptin improved hepatic steatosis induced by IR and IGF1R inhibition via a previously unknown mechanism that did not involve gluconeogenesis, lipogenesis, or inflammation, suggesting the non-canonical actions of DPP-4 inhibitors in the treatment of hepatic steatosis under insulin-resistant conditions.

Efficient synthesis, structure, and complexation studies of electron-donating thiacalix[n]dithienothiophene.

A series of thiacalix[n]dithiothiophenes (n=4-10) was prepared by a facile method and X-ray analysis was used to determine the molecular structures of square- (4-mer) and pentagonal-shaped macrocycles (5-mer). In the cyclic voltammograms, reversible multielectron redox processes, which are due to electronic delocalization, were observed at low oxidation potentials. The cyclic 4-mer acted as a "Janus-head" cavitand for two C60 molecules, whereas the 5- and 6-mer formed stable 1:1 complexes with C60 .

Solvent-free transformation of protic ionic liquids into zwitterions.

We synthesized several protic ionic liquids (ILs) composed of onium cations and the (trifluoromethylsulfonyl)(vinylsulfonyl)amide anion. The addition of a base catalytically facilitated their transformation into zwitterions (ZIs) under solvent-free conditions, which is a convenient method for synthesizing ZIs from ILs.

Controlling Ionic Conductivity in Organometallic Ionic Liquids through Light-Induced Coordination Polymer Formation and Thermal Reversion.

Owing to their high ionic conductivity and negligible vapor pressure, ionic liquids (ILs) find applications in various electronic devices. However, fabricating IL-based photocontrollable devices remains a challenge. In this study, we developed organometallic ILs with reversible light- and heat-controlled ionic conductivities for potential use in tunable devices. The physical properties and stimulus responses of ILs containing a cationic sandwich Ru complex with two coordinating substituents were investigated. UV photoirradiation of these ILs triggered cation photodissociation, resulting in their transformation into viscoelastic coordination polymers wherein the coordinating substituents bridged the Ru centers. Owing to the anion coordination, salts with coordinating anions such as CF3SO2NCN-, B(CN)4-, and BF2(CN)2- exhibited faster response and higher conversion than those with noncoordinating anions including (FSO2)2N- and (CF3SO2)2N-. All photoproducts reverted to their original ILs upon heating, except for the photoproduct of the BF2(CN)2 salt, which decomposed upon heating. Light- and heat-induced reversible changes occur in most cases between the high-ionic-conductive IL state and low-ionic-conductive coordination polymer state. Unlike previously reported ILs with three or one cation substituent, the current ILs exhibited both high reactivity and large ionic conductivity changes.

Immediate Impact of Switching from Dipeptidyl Peptidase 4 (DPP4) Inhibitors to Low-Dose (0.3 mg) Liraglutide on Glucose Profiles: A Retrospective Observational Study.

INTRODUCTION: As treatment agents for diabetes, liraglutide is a long-acting glucagon-like peptide 1 receptor agonist, and dipeptidyl peptidase 4 (DPP4) inhibitors are widely used because of their safety and tolerability. Regular treatment with liraglutide has been reported to significantly reduce blood glucose levels, but the impact of low-dose (0.3 mg) liraglutide on blood glucose levels immediately after treatment switching from a DPP4 inhibitor remains unknown. METHODS: We conducted a single-arm, retrospective, observational study in 55 inpatients with type 2 diabetes (T2D) to investigate the changes (Δ) in their blood glucose levels at six time points (6-point) from the day before (day -1) to the day after (day 1) by switching the antidiabetic treatment from a DPP4 inhibitor to liraglutide 0.3 mg (low-dose liraglutide) once daily. We also attempted to identify factors associated with the blood glucose-lowering effects of liraglutide. RESULTS: The median values of the changes in fasting, preprandial, and postprandial blood glucose levels and the fluctuations in the blood glucose levels expressed as the standard deviation of the 6-point blood glucose levels were significantly lower on day 1 than on day -1 (P < 0.05, P < 0.0001, P < 0.0001, P < 0.01, respectively); there were no cases of severe hypoglycemia. The Δ blood glucose levels were not associated with the baseline serum hemoglobin A1c values or with any markers of the insulin secreting capacity. There were no associations between the previously used blood glucose-lowering drug and the Δ blood glucose levels. CONCLUSION: Switching from a DPP4 inhibitor to low-dose (0.3 mg) liraglutide once daily significantly reduced the blood glucose levels and excursions of the blood glucose levels even from the very day after the treatment switch, with no serious adverse events.

Preparation and evaluation of sustained release formulation of PLGA using a new injection system based on ink-jet injection technology.

We developed a method for the preparation of PLGA particles exhibiting long-term sustained-release of entrapped drugs. The fine droplet drying (FDD) technology using a new injection system based on ink-jet injection technology was adapted as the preparation method. PLGA microspheres containing TRITC-dextran, acetaminophen, and albumin as model drugs were prepared by the FDD technology. The resultant microspheres were uniform in size, with average particle sizes ranging from 16.3 to 33.0 µm and SPAN factors ranging from 0.49 to 0.77. The encapsulation efficiency of drugs showed high values ranging from 75 to 99 wt% of the total amount of water-soluble drug contained in the particles. In an investigation of the optimal operation conditions of the FDD technology, the dew point temperature of the dryer air stream was found to be an important factor for controlling the initial burst of the prepared particles. The TRITC-dextran-containing PLGA microspheres were confirmed to exhibit long-term sustained release for about 90 days, and the mechanism was found to be PLGA degradation rate-limiting. Based on these results, we concluded that long-term sustained-released PLGA particles can be prepared by using FDD technology under a suitable drying condition for controlling the initial burst.

A Disproportionality Analysis of the Adverse Effect Profiles of Methimazole and Propylthiouracil in Patients with Hyperthyroidism Using the Japanese Adverse Drug Event Report Database.

Background: Antithyroid drugs (ATDs) are frequently used to achieve euthyroidism in patients with hyperthyroidism. ATDs cause characteristic common and rare adverse events; however, comprehensive comparisons between methimazole (MMI) and propylthiouracil (PTU) in terms of adverse events are limited. Methods: In this study, we thoroughly explored adverse events in association with MMI and PTU use with a disproportionality analysis using the Japanese Adverse Drug Event Report (JADER) database and evaluated the prevalence of MMI and PTU prescriptions using the National Database of Health Insurance Claims and Specific Health Checkups (NDB) Open Data Japan. We analyzed 3271 cases of MMI use and 1029 cases of PTU use with respect to 9789 preferred terms (PTs) for adverse events registered in the JADER database by calculating and comparing reporting odds ratios (RORs). Results: We found that 8 PTs, including agranulocytosis (p < 0.0001, 4.01-fold), aplasia cutis congenita (p < 0.0001, 123.22-fold), and exomphalos (p = 0.0002, 22.17-fold), demonstrated significantly higher RORs (more than 4-fold) for MMI use than for PTU use. Nineteen PTs, including anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (p < 0.0001, 29.84), rapidly progressive glomerulonephritis (p < 0.0001, 6.44), and pulmonary alveolar hemorrhage (p < 0.0001, 7.77), had RORs for PTU use more than four times those for MMI use. NDB Open Data Japan showed more frequent PTU prescriptions than MMI prescriptions for women of reproductive age. Conclusions: This large-scale study confirmed that a variety of congenital malformations were identified as having significantly high RORs for MMI use, while diseases related to ANCA-associated vasculitis were specific to PTU.

Protective Effects of Imeglimin and Metformin Combination Therapy on ß-Cells in db/db Male Mice.

Imeglimin and metformin act in metabolic organs, including ß-cells, via different mechanisms. In the present study, we investigated the impacts of imeglimin, metformin, or their combination (Imeg + Met) on ß-cells, the liver, and adipose tissues in db/db mice. Imeglimin, metformin, or Imeg + Met treatment had no significant effects on glucose tolerance, insulin sensitivity, respiratory exchange ratio, or locomotor activity in db/db mice. The responsiveness of insulin secretion to glucose was recovered by Imeg + Met treatment. Furthermore, Imeg + Met treatment increased ß-cell mass by enhancing ß-cell proliferation and ameliorating ß-cell apoptosis in db/db mice. Hepatic steatosis, the morphology of adipocytes, adiposity assessed by computed tomography, and the expression of genes related to glucose or lipid metabolism and inflammation in the liver and fat tissues showed no notable differences in db/db mice. Global gene expression analysis of isolated islets indicated that the genes related to regulation of cell population proliferation and negative regulation of cell death were enriched by Imeg + Met treatment in db/db islets. In vitro culture experiments confirmed the protective effects of Imeg + Met against ß-cell apoptosis. The expression of Snai1, Tnfrsf18, Pdcd1, Mmp9, Ccr7, Egr3, and Cxcl12, some of which have been linked to apoptosis, in db/db islets was attenuated by Imeg + Met. Treatment of a ß-cell line with Imeg + Met prevented apoptosis induced by hydrogen peroxide or palmitate. Thus, the combination of imeglimin and metformin is beneficial for the maintenance of ß-cell mass in db/db mice, probably through direct action on ß-cells, suggesting a potential strategy for protecting ß-cells in the treatment of type 2 diabetes.