Effects of Complement Regulators and Chemokine Receptors in Type 2 Diabetes.

CD55 and CD59 are complement regulatory proteins suggested to be related with progression of diabetes and its complications. The stromal cell-derived factor 1 (SDF-1) and C-X-C chemokine receptor type 4 (CXCR-4) are chemokine proteins. We aimed to investigate the relation of CD55 and CD59 expression levels and polymorphisms of SDF-1 and CXCR-4 with type 2 diabetes mellitus (T2DM) and its complications. Seventy-five T2DM patients and 73 controls were enrolled. Expression levels of CD55 and CD59 were measured by FACS Calibur; qRT-PCR was used to determine SDF-1 and CXCR-4 gene polymorphisms. CD55 and CD59 expressions in patients with nephropathy, retinopathy and cardiovascular disease were significantly lower than controls. Frequency of CXCR-4 T allele carrying was high in patients and created 1.6 fold risk for the disease (p = .07). CXCR-4 a allele carriers had decreased nephropathy; although there was no statistical significance in carrying CXCR-4 T allele, presence of nephropathy was approximately 2 times higher (p = .254). The nephropathy risk increased 10-fold in CXCR-4 TT genotype carriers (p = .02). All SDF-1 CC genotype carriers had retinopathy, so, it was considered that the CC genotype was effective in retinopathy development (p = .031). For the presence of cardiovascular disease, significant difference was observed for SDF-1 genotypes. Increased cardiovascular risk of 5- and 1.9-fold in SDF-1 T (p = .007) and CXCR-4 T (p = .216) allele carriers, respectively, was observed. We suggest that CD55 and CD59 protein levels and SDF-1 and CXCR-4 have predictive importance in process, complications and tendency of T2DM.

The synergistic effect of ω3 and Vit D3 on glycemia and TNF-α in islet transplantation.

The current treatment of type 1 diabetes consists of insulin administration. Transplantation of islets of Langerhans is considered very favorable because the full effect of insulin treatment cannot be obtained in severe cases. Although agents such as omega-3 (ω3) and vitamin D3 (Vit D3) are known to contribute to the success of islet allo-transplantation (ITX), in this study we aimed to experimentally determine their effects on glycemia and TNF-α production. Wistar albino rats, which were used as recipients, were given ω3, Vit D3, and islets by gavage, and intraperitoneal- and intraportal injections, respectively. Daclizumab (DAC) was used for immunosuppression. Glycemia levels decreased in rats treated with ω3 and vit D3. TNF-α increased in all groups due to application of STZ. After ITX (day +1), the weakest increase was observed in the ω3 + Vit D3 group. In the ITX+DAC group, compared with that of ITX only, DAC was shown to decrease levels of TNF- α following ITX, only in control group, however, similar levels of TNF-α were observed in other groups. The values in the treated groups were already lower than those of the controls in the ITX group and also remained almost equal in the ITX+DAC group. We suggest that the use of ω3 and Vit D3 together will improve the pro-inflammatory aspect encountered during and after ITXs, and contribute to the reduction of the dose of immunosuppressants in these procedures.

Activity of dental pulp cells in semisolid 3D cultures initiated by transforming growth factor-ß1 and bone morphogenetic protein 2, 4.

The intention of this study was to investigate the effect of modified 3D culture conditions on dental pulp cells (DPCs). DPCs were isolated from extracted primary molar, premolar, and wisdom teeth. Tooth samples were divided into three groups as control group; plated into methyl cellulose medium without any supplementation, growth factor (GF) group; supplemented with bone morphogenetic proteins (BMP2, BMP4), transforming growth factor—β1 (TGF—β1) and growth factor+conditioned medium (GF+CM) group; supplemented with both growth factors and pulp conditioned medium. The DPCs were tested for colony forming ability, proliferation capacity and morphology. The highest colony forming ability was detected in the GF and GF+CM groups of DPCs isolated from wisdom teeth. The proliferation capacity was higher in GF+CM group of DPCs isolated from primary molars, and in GF and GF+CM groups of DPCs isolated from wisdom teeth. Scanning electron microscope (SEM) observation of the wisdom teeth samples showed cell—cell interactions in the GF and GF+CM groups. Our results indicate that growth factors and pulp conditioned medium in methyl cellulose culture created proper environment to follow the behavior of dental cells three—dimensionally.

Organ specific autoantibodies in preclinical and early clinical type 1 diabetes in Turkey.

Type 1 diabetes mellitus (T1D) patients (G1; n=73) and first degree relatives with islet cell antibody (ICA) values of >or=10 JDF u twice or >or=20 JDF u one and loss of FPIR (G2; n=18) were screened for two other autoantibodies, anti-glutamic acid decarboxylase (GADA) and insulin autoantibodies (IAA), and for other organ-specific autoantibodies, anti-gastric parietal cell (anti-PCA) and anti-thyroid peroxidase (anti-TPO) as well. The two control groups consisted of healthy subjects (G3; n:55 and G4; n:13). In G1, positivity of ICA, GADA, IAA, anti-TPO and anti-PCA were 63%, 75.1%, 27.4%, 17.8% and 8.2%, respectively. In G2, positivity for GADA, IAA, anti-TPO and anti-PCA were 55.6%, 11.1%, 16.7% and 11.1%, respectively. None of the anti-TPO or anti-PCA positive cases had clinical or laboratory thyroid disease or pernicious anemia. Other organ specific antibodies, in case they accompany GADAand/or IAA in high risk individuals, result in higher risk for T1D. Moreover, this condition may indicate future potential for developing thyrogastric autoimmune diseases. In conclusion; autoantibodies are markers for autoimmune destruction in T1D, and for identification of subjects at risk for disease. Even at the time of diagnosis of T1D, screening for thyrogastric autoimmunity might be recommended for early detection of the relevant diseases.

Peritransplant and long-term secretion of interleukin-1beta in cyclosporine treated syngeneic rats allografted with islets of langerhans.

Interleukin-1beta (IL-1beta) is one of the proinflammatory cytokines that may mediate primary nonfunction of islets of Langerhans, limiting the success of allogeneic transplantation. The aim of this study was to assess differences between the secretion of IL-1beta as well as glycemia in peri- and long-term periods of intraportal islet allo-transplantation with or without cyclosporine (CyA) immunosuppression. Inbred Wistar albino rats were transplanted intraportally with rat islets isolated by collagenase digestion. The two recipient groups (6 rats/group) were: group 1, control, islet transplantation (ITX) without any treatment and group 2, CyA-treated via the femoral muscle on days -1, 0, +1, and +2. Serum IL-1beta (pg/mL) levels were measured by ELISA on days 0 (pre-ITX), +1, +2, and +195. Tail vein blood was used to evaluate glycemia (mg/dL). No major differences were observed in IL-1beta secretion on days 0, +1, or +195 between the groups. Immunosuppressive treatment produced significantly lower secretion in group 2 (P < .002) on day +2. Significantly greater secretions were detected at days +195, +1, and +195 compared to days 0, +2, and +2, respectively (P < .002; P < .008; P < .002). Positive correlations were observed between IL-1beta levels on days +1 and +2 (r = 0.845, P < .034). The mean values in groups 1 and 2 on days 0, +1, and +2 were 140.6 +/- 4.62 vs 119.1 +/- 12.12, 73.1 +/- 19.59 vs 88.3 +/- 14.08, 106.5 +/- 13.79 vs 92.5 +/- 15.8, respectively. No animal in group 1 displayed glycemia while three group 2 animals did at day +195. However, a negative correlation was found between IL-1beta on day 0 and glycemia on day +195 (r = -0.999, P < .026). Our results suggest that IL-1beta secretion, which is detrimental for islet engraftment, decreases at peritransplant day +2, but is upregulated during long-term graft survival both in controls and in CyA-treated recipients.

A modified automated method for isolation of viable pancreatic islets in laboratory animals.

To achieve successful islet transplantation, a high viability is required. For this reason an automated method including two chambers: one for islets isolation and one for recirculation and collection was developed. Recently, we produced a modified version of this work by building a similar system of glass where marbles were not used for disaggregation, and the pancreatic tissue had to be disrupted mechanically before the digestion phase. By using the reconfigured system, we obtained 260 +/- 20 islets from each Wistar albino rat (weighing 220 to 240 g) pancreas. Islets were observed at 35 minutes after the start of perfusion (closed circuit) and the optimum time to stop the isolation determined to be 40 minutes based upon islets viability. Although the present system is configured for islet isolation from small laboratory animals (rat, mouse), we have also obtained thousands of islets at 25 minutes after treatment of a 0.5-g piece of pig pancreas. Compared to the time-consuming manual method usually used for islet isolation from small laboratory animals, the new technique is economic, easy to use, and does not reduce islets viability.

Effect of human somatotropin hormone on cultured rat islets.

Because growth hormone (GH) improves the insulin secretion capacity of isolated human fetal islets in vitro, we sought to show that it positively influences isolated rat islets. Islets isolated from Wistar albino rats by a modified automated system were cultured in media containing 87% RPMI 1640, 10% FCS, 2% antibiotic-antimycotic, and 1% L-glutamine for 12 +/- 2 days. The cultured islets were divided into two groups: growth hormone negative (Group I) and growth hormone positive (Group II). On the 5th day we observed a decrease in the islet cell counts in both groups (Group I 28% versus Group II 45%). On the 10th day, the decrease continued in the GH-negative group (59%), while the count remained stable in the GH-positive group. The viability of rat islets was determined by fluorescein diacetate (FDA) plus propidium iodide (PI) staining. In comparison to the peripheral green, central orange-red staining pattern of Group I islets upon fluorescent microscopy, Group II showed more compact islets. Cultured islets seemed to be brighter than those without GH in the cultured islets. In conclusion, we observed that 2 weeks of incubation in the presence of GH acts positively on cultured rat islets for both their amount and their viability.

Doxazosin, but not amlodipine decreases insulin resistance in patients with chronic renal failure: a prospective, randomized-controlled study.

Insulin resistance (IR) in chronic renal failure (CRF) is well-known. In this randomized-controlled study, we aimed to compare the effect of doxazosin and amlodipine on IR in patients with CRF. Fifteen patients with CRF (male/female: 5/10, mean age: 46 +/- 13 years) and 9 controls (male/female: 3/6, mean age: 35 +/- 8 years) were included. Patients and controls had no family history of diabetes mellitus. Homeostasis model assessment (HOMA) was calculated as a marker of IR. Patients were grouped randomly to doxazosin (n = 8; 2-4 mg/day) and amlodipine (n = 7; 5-10 mg/day) arms. Baseline biochemical analysis (fasting serum glucose, BUN, creatinine, uric acid, cholesterol and cholesterol subgroups) and parameters related with insulin metabolism (insulin, C peptide, HOMA) were similar between amlodipine and doxazosin groups. There was no difference in age, gender and body mass index among study groups. The follow-up time was 12 weeks. Patients with CRF had higher HOMA (1.83 +/- 0.55 vs 1.00 +/- 0.36, p = 0.001), fasting insulin (8.06 +/- 1.98 vs 4.46 +/- 1.31 IU/l, p < 0.001) and serum triglyceride levels (197 +/- 136 vs 112 +/- 67 mg/dl, p = 0.04) as compared to controls. Serum HDL cholesterol levels were significantly lower in patients with CRF than controls (40 +/- 10 vs 57 +/- 14 mg/dl, p = 0.02). HOMA significantly decreased after doxazosin (1.91 +/- 0.45 vs 1.41 +/- 0.21, p = 0.02), however, no difference was found after amlodipine. Also, fasting insulin levels were decreased after a 12-week doxazosin therapy from 8.17 +/- 1.22 vs 6.58 +/- 0.84 IU/l, p = 0.02), but no change was seen after amlodipine. Lipid parameters did not significantly change during the study period in 2 groups. No adverse effect requiring drug discontinuation was observed during the 12-week period in the study groups. In conclusion, doxazosin decreases IR in patients with CRF, whereas amlodipine has no effect. This may be of advantage in the treatment of hypertension in this group of patients for preventing some long-term complication of IR.

Influence of insulin resistance on total renin level in normotensive women with polycystic ovary syndrome.

OBJECTIVE: To evaluate the influence of insulin resistance on the plasma total renin level in normotensive women with polycystic ovary syndrome (PCOS). DESIGN: Prospective, controlled study. SETTING: University hospital. PATIENT(S): Twenty-five normotensive women with PCOS were compared with 11 normotensive control women with regular cycles and no features of PCOS. INTERVENTION(S): Clinical, ultrasonographic, and hormonal findings were used to define PCOS. Insulin resistance was estimated by continuous infusion of glucose with model assessment in the early follicular phase. MAIN OUTCOME MEASURE(S): Plasma levels of total renin and angiotensin II and serum levels of gonadotropins, DHEAS, total T, free T, 17 alpha-hydroxyprogesterone, and PRL were determined. RESULT(S): Plasma concentrations of angiotensin II were similar in the PCOS group and the control group. The concentration of total renin in plasma was higher in women with PCOS than in healthy women independent of insulin resistance. The sensitivity and specificity of the plasma total renin level to diagnose women with PCOS were calculated as 80% and 71.4%, respectively. CONCLUSION(S): The plasma total renin level is higher in normotensive women with PCOS than in healthy women independent of insulin resistance.