Probiotic reduces bacterial translocation in type 2 diabetes mellitus: A randomised controlled study.

Gut bacterial translocation to the blood may play an important role in the development of insulin resistance in type 2 diabetes. Here, we performed an interventional randomised control study to investigate whether probiotics could reduce bacterial translocation and cause changes in the gut microbiota. Seventy Japanese patients with type 2 diabetes were randomised to two groups: the probiotic group drank Lactobacillus casei strain Shirota-fermented milk, while the control group ingested no probiotics. The trial was conducted for 16 weeks. At baseline, 8 and 16 weeks, the gut microbiota composition in feces and blood, fecal organic acids, and other biochemical parameters were measured. At the end of the study, the fecal counts of the Clostridium coccoides group and Clostridium leptum subgroup in the probiotic group were significantly higher than in the control group. As expected, the fecal counts of total Lactobacillus were significantly higher in the probiotic group. Intriguingly, the total count of blood bacteria was significantly lower in the probiotic group. However, fecal organic acids were comparable between the two groups. Our results showed that probiotic administration reduced bacterial translocation and altered the gut microbiota in Japanese patients with type 2 diabetes mellitus.

Comparison of effects of azelnidipine and trichlormethiazide in combination with olmesartan on blood pressure and metabolic parameters in hypertensive type 2 diabetic patients.

UNLABELLED: Aims/Introduction: Angiotensin II type 1 receptor blockers (ARB) are regarded as first-line treatment for type 2 diabetes with hypertension. However, lowering blood pressure to the target level often requires more than one antihypertensive agent as recommended by the guideline. In this open-label, prospective, crossover clinical trial, we compared the effects of combination treatment of ARB with a calcium channel blocker (CCB) or with a low-dose thiazide diuretic on blood pressure (BP) and various metabolic parameters in hypertensive patients with type 2 diabetes. MATERIALS AND METHODS: A total of 39 Japanese type 2 diabetics with hypertension treated with olmesartan (20 mg/day) for at least 8 weeks were recruited to this study. At study entry, treatment was switched to either olmesartan (20 mg/day)/azelnidipine (16 mg/day) or olmesartan (20 mg/day)/trichlormethiazide (1 mg/day) and continued for 12 weeks. Then, the drugs were switched and treatment was continued for another 12 weeks. We measured clinical blood pressure and various metabolic parameters before and at the end of each study arm. RESULTS: Compared with the olmesartan/trichlormethiazide treatment, treatment with olmesartan/azelnidipine achieved superior clinical blood pressure and pulse rate control. In contrast, the treatment with olmesartan/trichlormethiazide resulted in increased HbA1c, serum uric acid and worsening of estimated glomerular filtration rate, though there were no differences in other metabolic parameters including urine 8-hydroxy-2'-deoxyguanosine, C-reactive protein and adiponectin between the two treatments. CONCLUSIONS: Our results show that the combination of ARB with azelnidipine is more beneficial with regard to blood pressure control and metabolic outcome than the combination of olmesartan with low dose trichlormethiazide. This trial was registered with UMIN clinical trial registry (no. UMIN000005064). (J Diabetes Invest, doi: 10.1111/j.2040-1124.2011.00135.x, 2011).

Superior thyroid artery mean peak systolic velocity for the diagnosis of thyrotoxicosis in Japanese patients.

Thyrotoxicosis with diffuse thyroid disease can be caused by Graves' disease (GD) or destructive thyroiditis (DT). Optimal treatment of the underlying condition requires a prompt and accurate method for the diagnosis of thyrotoxicosis. This study evaluated measurement of the mean peak systolic velocity of the superior thyroid artery (STA-PSV) by ultrasonography in detecting thyrotoxicosis in Japanese patients. We recruited 44 patients with untreated GD, 13 with DT, 55 with treated GD, and 49 subjects without thyroid disease. Blood samples were taken to analyze thyroid function and STA-PSV was measured by ultrasonography. The mean STA-PSV was the highest in the untreated GD group, followed by treated GD patients and then those with DT. Receiver operating characteristic curves of the STA-PSV values demonstrated that the area under the curve required discriminating untreated GD from DT was 0.941. The optimal sensitivity and specificity were 83.7% and 92.3%, respectively, using 45 cm/sec as the cutoff value. In conclusion measurement of STA-PSV by ultrasonography is useful for the diagnosis of thyrotoxicosis in Japanese patients.

Superior thyroid artery mean peak systolic velocity for the diagnosis of thyrotoxicosis in Japanese patients.

Thyrotoxicosis with diffuse thyroid disease can be caused by Graves' disease (GD) or destructive thyroiditis (DT). Optimal treatment of the underlying condition requires a prompt and accurate method for the diagnosis of thyrotoxicosis. This study evaluated measurement of the mean peak systolic velocity of the superior thyroid artery (STA-PSV) by ultrasonography in detecting thyrotoxicosis in Japanese patients. We recruited 44 patients with untreated GD, 13 with DT, 55 with treated GD, and 49 subjects without thyroid disease. Blood samples were taken to analyze thyroid function and STA-PSV was measured by ultrasonography. The mean STA-PSV was the highest in the untreated GD group, followed by treated GD patients and then those with DT. Receiver operating characteristic curves of the STA-PSV values demonstrated that the area under the curve required discriminating untreated GD from DT was 0.941. The optimal sensitivity and specificity were 83.7% and 92.3%, respectively, using 45 cm/sec as the cutoff value. In conclusion measurement of STA-PSV by ultrasonography is useful for the diagnosis of thyrotoxicosis in Japanese patients.

Combined expression of transcription factors induces AR42J-B13 cells to differentiate into insulin-producing cells.

Neurogenin 3 (Ngn3) is a transcription factor that regulates an initial step of differentiation from uncommitted pancreatic progenitors into endocrine cells. Additional transcription factors are required for complete differentiation into mature pancreatic beta cells. In this study, we established an in vitro model system of beta-cell differentiation by adenovirus-mediated expression of several transcription factors in AR42J-B13 cells, a pancreatic progenitor-like cell line derived from exocrine pancreas. Exogenous expression of Ngn3 in AR42J-B13 cells induced expression of Nkx2.2, Pax4, and Pax6, which are all essential for beta-cell differentiation in mouse embryos. However, Ngn3 did not induce more downstream regulators of beta-cell differentiation, Nkx6.1 and Maf A. Coexpression of Nkx6.1 and Ngn3 induced endogenous expression of the insulin 2 gene, while coexpression of Maf A and Ngn3 induced both insulin 1 and 2 genes in AR42J-B13 cells. Our data demonstrated that Ngn3 expressed together with Nkx6.1 or MafA induces AR42J-B13 cells to differentiate into insulin-producing cells, supporting the use of these cells as a model system for studying beta-cell differentiation in vitro.

Activin A-induced expression of PAX4 in AR42J-B13 cells involves the increase in transactivation of E47/E12.

Pax4 is a paired-homeodomain containing transcriptional factor that controls the differentiation of pancreatic beta cells. The aim of this study was to investigate the mechanism of PAX4 expression by activin A. By reporter gene analysis using AR42J-B13 cells, in which treatment with activin A induced PAX4 mRNA expression, we identified that a short sequence located approximately 1930 bp upstream of the transcriptional start site is essential for activin A induced PAX4 promoter activation. This region contains an E box and binding sites for hepatocyte nuclear factor (HNF)-1alpha. Mutation introduced in each binding site markedly reduced activin A responsiveness. It has been reported that HNF-1alpha synergizes with basic helix-loop-helix (bHLH) proteins in activating the PAX4 promoter, and we demonstrated that activin A strongly enhanced the functional activity of E47/E12 without the increase in its binding ability. In addition, suppression of E47/E12 expression in AR42J-B13 cells using siRNA oligonucleotides results in the significant decrease in the intrinsic activin A-induced PAX4 expression. Our results suggest that activin A enhances PAX4 expression by enhanced transactivation of E47/E12 proteins and might result in a cumulative transactivation of the promoter.

Therapeutic efficacy of mitiglinide combined with once daily insulin glargine after switching from multiple daily insulin regimen of aspart insulin and glargine in patients with type 2 diabetes mellitus.

Mitiglinide is novel class of rapid-acting insulin secretagogues, which have been widely used alone or in combination with other oral hypoglycemic drugs to improve postprandial hyperglycemia in early type 2 diabetes. While mitiglinide enhances postprandial requirement of insulin, the efficacy of mitiglinide combined with insulin has yet to be established. We investigated the efficacy of mitiglinide combined with insulin glargine, the first soluble insulin analog that has a flat and prolonged effect. After control with the intensive regimen (daily aspart insulin and glargine), 30 inpatients with type 2 diabetes were switched to premeal mitiglinide combined with once daily insulin glargine (mitiglinide regimen), and daily profiles of blood glucose level were compared under each regimen. Fifteen patients showed similar control of hyperglycemia with mitiglinide regimen and intensive insulin regimen, assessed by M value (<32), while the remaining 15 showed worsening under the mitiglinide regimen. The patients who were well controlled with mitiglinide regimen were significantly younger (51.9 +/- 16.0 years, p<0.005) and heavier (body mass index: 25.7 +/- 3.3 kg/m(2), p<0.05) than those who were not (67.9 +/- 8.7 and 23.0 +/- 3.1, respectively). Moreover, insulin doses of aspart per body weight were significantly fewer in effective group than in ineffective group. Duration of diabetes was shorter in the effective group, albeit insignificantly. Previous treatment before starting intensive insulin regimen, such as insulin and sulfonylurea, was not different between the two groups. Our results suggest that mitiglinide plus insulin glargine combination therapy is useful for lowering both fasting and postprandial hyperglycemia in a subpopulation of type 2 diabetes. The long-term effects of such treatment need to be established in future studies.

p38 MAPK is involved in activin A- and hepatocyte growth factor-mediated expression of pro-endocrine gene neurogenin 3 in AR42J-B13 cells.

Neurogenin3 (ngn3) is a transcription factor that is essential for the differentiation of pancreatic endocrine cells. To investigate the signaling pathway that regulates ngn3 expression, we used AR42J-B13 cells as a model of the differentiation of pancreatic islets. In these cells, treatment with activin A and hepatocyte growth factor (HGF) induced the expression of ngn3. Reporter gene analysis using human ngn3 gene (NEUROG3) promoter fragments of various lengths identified the region between -402 and -327 bp of NEUROG3 as an activin A- and HGF-responsive DNA sequence. This DNA sequence normally functions as a repressor in AR42J-B13 cells, but treatment with activin A and HGF negates the repressor activity. Interestingly, function of the activin A- and HGF-responsive sequence was not influenced by the overexpression of the Smad inhibitory factor, Smad7. Instead, activin A and HGF activation was inhibited by overexpression of a dominant-negative mutant of transforming growth factor-beta-activated kinase 1 (TAK1), or mitogen-activated protein kinase kinase 3 (MKK3), or by treatment with a p38 MAPK-specific inhibitor, SB203580. Activin A and HGF function through the TAK1-MKK3-p38 MAPK pathway to relieve transcription repressors located between -402 and -326 bp on the NEUROG3 promoter, and consequently activate ngn3 expression and endocrine differentiation of AR42J-B13 cells.

RANTES promoter genotype is associated with diabetic nephropathy in type 2 diabetic subjects.

OBJECTIVE: To evaluate the effect of RANTES gene promoter polymorphism and RANTES receptor (CCR5 gene) promoter polymorphism on diabetic nephropathy in Japanese type 2 diabetic subjects. RESEARCH DESIGN AND METHODS: A total 616 Japanese subjects with type 2 diabetes were recruited. Polymorphisms of -28 C/G and -403 G/A in the RANTES gene promoter region, and of 59029 G/A in the CCR5 gene promoter region were detected by PCR-RFLP (restriction fragment length polymorphism). The association of these genotypes with nephropathy was analyzed. RESULTS: While the RANTES -403 genotype showed no association with nephropathy, the frequency of the -28G allele was significantly higher in the DN2 group (urinary albuminuria-to-creatinine ratio [ACR] >or=300 mg/g creatinine, serum creatinine <2.0 mg/dl) than in the DN0 (ACR <30 mg/g creatinine) and DN1 (ACR >or=30 mg/g creatinine and <300 mg/g creatinine) groups. The frequency of a RANTES -28G-positive genotype (C/G or G/G) was higher in the DN2 group than in the DN0 and DN1 groups (34% vs. 25 and 20%, P = 0.0268, chi(2) = 4.905), and the frequency of a CCR5 59029 A-positive genotype (G/A or A/A) was higher in the DN1 and DN2 groups than in the DN0 group (84 and 85% vs. 76%, P = 0.0123, chi(2) = 6.269). Discriminant analysis showed that the RANTES -28G-positive genotype and CCR5 59029A-positive genotype were independently associated with nephropathy. The percentage of macroalbuminuria was twofold higher in the subjects having -28G or 59029A and threefold higher in the subjects having -28G and 59029A than in the subjects without -28G and 59029A. CONCLUSIONS: The RANTES promoter -28G genotype and CCR5 promoter 59029A genotype may be independent risk factors for diabetic nephropathy in patients with type 2 diabetes and may have an additive effect on nephropathy.