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ABSTRACTAim:To substantially increase awareness, treatment and effective control of the metabolic syndrome (MetS) and its components.Subjects and methods:This is a pilot best practice implementation enhancement programme to reduce the estimated cardiovascular disease (CVD) risk in 628 MetS patients with or without diabetes or CVD by improving quality of care. A baseline visit was followed by action to improve adherence to lifestyle advice and drug treatment for CVD risk factors by physicians specifically trained to implement guidelines. Finally, after 6 months, a single-page form was completed, showing if patients were at CVD risk factor target. If not, there was an analysis of the reason why.Results:The programme was effective in improving utilization of evidence-based treatment in 628 MetS patients. There was a substantially greater patient perception of MetS, an enhancement in compliance with lifestyle advice and increased prescription of evidence-based medication, leading to a 48 (p < 0.0001) improvement in estimated CVD risk. There was a substantial increase in the number of subjects on target for specific CVD risk factors.Conclusions:This is the first study to increase adherence to multiple interventions for all MetS components on an outpatient basis, in both primary care and teaching hospital settings. Physician and patient education, distribution of printed guidelines and brochures, and completion of a single-page form motivated both physicians and patients to achieve multiple CVD risk factor guideline goals. The absence of a control group is a limitation of this study. Further work is also needed to establish if the improvements observed are sustained on a long-term basis. [ABSTRACT FROM AUTHOR]

Background: Invariant Natural Killer T (iNKT) cells belong to innate immunity and combine T-cell receptor specificity with Natural Killer surface markers. They can produce cytokines immediately after stimulation and direct immunity to either Th1 or Th2 cytokine production. iNKT cells participate in a variety of immune responses, such as microbial infections, autoimmunity, and cancer. Type 2 Diabetes Mellitus (T2DM) has been associated with activated innate immunity and certain cytokine profile during microbial infections. This study aimed to evaluate whether iNKT cells have a role in the immune response of T2DM patients during infections with gram-negative bacteria. Method: The T2DM group consisted of patients (n =11) who had a diagnosis of T2DM for at least six months and febrile illness for three days, while the control group consisted of patients (n =11) who had not T2DM, but were febrile for three days. All patients were infected by gram-negative bacteria. Physical examination was performed, and peripheral blood was drawn on days three and six of febrile illness. Flow cytometry was utilized for iNKT cell identification with monoclonal antibodies Phycoerythrin (PE) - Cyanin (CY) 5 anti-Human CD3, Fluorescein isothiocyanate (FITC) anti-Human CD4, PE anti-human invariant NKT T-Cell Receptor. For intracellular staining, we used Alexa Fluor anti-Human interferon-? (IFN-?) and Allophycocyanin (APC) anti-human interleukin-4 (IL-4). The variables processed were: CD3+IL-4+iNKT+, CD4+IL-4+iNKT+, CD3+IFN?+iNKT+, CD4+IFN?+i???+, CD3+iNKT+, CD4+iNKT+,CD3+IL4+, CD4+IL-4+, CD3+IFN?+, CD4+IFN?+ on days three and six of febrile illness (CD3+, CD4+: T lymphocyte surface markers, iNKT+: invariant Natural Killer T- Cell Receptor, IL4+: interleukin 4, IFNγ+: interferon γ). Results: Comparisons between T2DM patients and controls revealed no statistically significant difference in any of the study's variable. Regarding within T2DM patients comparisons CD4+IL4+iNKT+, CD3+IL4+iNKT+, CD4+IFN+iNKT+, CD3+IFN+iNKT+, and CD3+iNKT+ decreased, whereas CD3+IL4+ was increased at day six compared to day three. Within control group CD4+IL4+iNKT+, CD3+IL4+iNKT+, and CD3+iNKT+ were decreased, whereas CD4+IFN+, CD3+IFN+ were increased at day six compared to day three. Conclusion: The absence of statistical difference between T2DM patients and controls implies that the role of iNKT cells is virtually the same in both groups of patients during the course of gram-negative infections and that there is no numerical variance of this cell population between the two groups. Despite the small sample size, we notice that all iNKT parameters (both IL4/IFN?) are suppressed in the T2DM group during the later phase, but only those concerning IL4+iNKT+ in the control group, suggesting that IFN? production remains elevated in the controls. A compensatory anti-inflammatory type-response could provide an explanation for the prevalence of IL4 production during the later phase of infection in T2DM and the sustained production of IFN? in controls. [ABSTRACT FROM AUTHOR]

Background: Invariant Natural Killer T (iNKT) cells belong to innate immunity and combine T-cell receptor specificity with Natural Killer surface markers. They can produce cytokines immediately after stimulation and direct immunity to either Th1 or Th2 cytokine production. iNKT cells participate in a variety of immune responses, such as microbial infections, autoimmunity, and cancer. Type 2 Diabetes Mellitus (T2DM) has been associated with activated innate immunity and certain cytokine profile during microbial infections. This study aimed to evaluate whether iNKT cells have a role in the immune response of T2DM patients during infections with gram-negative bacteria., Method: The T2DM group consisted of patients (n =11) who had a diagnosis of T2DM for at least six months and febrile illness for three days, while the control group consisted of patients (n =11) who had not T2DM, but were febrile for three days. All patients were infected by gram-negative bacteria. Physical examination was performed, and peripheral blood was drawn on days three and six of febrile illness. Flow cytometry was utilized for iNKT cell identification with monoclonal antibodies Phycoerythrin (PE) - Cyanin (CY) 5 anti-Human CD3, Fluorescein isothiocyanate (FITC) anti-Human CD4, PE anti-human invariant NKT T-Cell Receptor. For intracellular staining, we used Alexa Fluor anti-Human interferon-γ (IFN-γ) and Allophycocyanin (APC) anti-human interleukin-4 (IL-4). The variables processed were: CD3+IL-4+iNKT+ , CD4+IL-4+iNKT+, CD3+IFNγ+iNKT+, CD4+IFNγ+iΝΚΤ+, CD3+iNKT+, CD4+iNKT+ ,CD3+IL4+, CD4+IL-4+, CD3+IFNγ+, CD4+IFNγ+ on days three and six of febrile illness (CD3+, CD4+: T lymphocyte surface markers, iNKT+: invariant Natural Killer T- Cell Receptor, IL4+: interleukin 4, IFNγ+: interferon γ)., Results: Comparisons between T2DM patients and controls revealed no statistically significant difference in any of the study's variable. Regarding within T2DM patients comparisons CD4+IL4+iNKT+, CD3+IL4+iNKT+, CD4+IFN+iNKT+, CD3+IFN+iNKT+, and CD3+iNKT+  decreased, whereas CD3+IL4+ was increased at day six compared to day three. Within control group CD4+IL4+iNKT+, CD3+IL4+iNKT+, and CD3+iNKT+ were decreased, whereas CD4+IFN+, CD3+IFN+ were increased at day six compared to day three., Conclusion: The absence of statistical difference between T2DM patients and controls implies that the role of iNKT cells is virtually the same in both groups of patients during the course of gram-negative infections and that there is no numerical variance of this cell population between the two groups. Despite the small sample size, we notice that all iNKT parameters (both IL4/IFNγ) are suppressed in the T2DM group during the later phase, but only those concerning IL4+iNKT+ in the control group, suggesting that IFNγ production remains elevated in the controls. A compensatory anti-inflammatory type-response could provide an explanation for the prevalence of IL4 production during the later phase of infection in T2DM and the sustained production of IFNγ in controls. Hippokratia 2015; 19 (3): 231-234.

Melanocortin-4 receptor (MC4R) loss-of-function mutations are the commonest genetic cause of human monogenic obesity, so far. The contribution of MC4R coding mutations to severe obesity in the high-obesity prone Greek population has not been investigated to date. We determined the MC4R coding sequence of 510 obese and 469 lean control subjects of Greek origin, and we estimated the prevalence and the penetrance on obesity of MC4R loss-of-function mutations. The functional impact of novel nonsynonymous variants detected was investigated in vitro. We found two novel synonymous mutations (L23L and I102I), four nonsynonymous mutations (T112M, S127L, N274S, and S295L), and two polymorphisms (V103I and I251L) previously described in literature. We also detected a novel mutation (L207V) in a severely obese 69-year-old female patient, although the mutation did not cosegregate with obesity in the corresponding pedigree and had no functional consequences on MC4R protein function. Loss-of-function mutations represented 75% of all nonsynonymous rare mutations identified among lean carriers and only 25% among obese subjects (P = 0.0001). The prevalence of loss-of-function mutations was lower in the obese group than in lean control subjects (0.20 vs. 0.64%) but this difference was not significant. Therefore, the estimated penetrance of deleterious MC4R mutations was very low (6.3%) in heterozygous Greek carriers of MC4R loss-of-function mutations. Our data suggest that MC4R loss-of-function mutations are rare in the Greek population. MC4R genetic deficiency is unlikely to explain the high propensity to develop severe obesity in this specific population.

Objective: To assess the efficacy and safety of dipeptidyl peptidase-4 (DPP-4) inhibitors compared with metformin as monotherapy, or with other commonly used hypoglycaemic drugs combined with metformin, in adults with type 2 diabetes mellitus., Design: Systematic review and meta-analysis of randomised controlled trials., Data Sources: Medline, Embase, the Cochrane Library, conference proceedings, trial registers, and drug manufacturers' websites., Eligibility Criteria: Randomised controlled trials of adults with type 2 diabetes mellitus that compared a DPP-4 with metformin as monotherapy or with a sulfonylurea, pioglitazone, a glucagon-like peptide-1 (GLP-1) agonist, or basal insulin combined with metformin on the change from baseline in glycated haemoglobin (HbA(1c))., Data Extraction: The primary outcome was the change in HbA(1c). Secondary outcomes included the proportion of patients achieving the goal of HbA(1c) <7%, the change in body weight, discontinuation rate because of any adverse event, occurrence of any serious adverse event, all cause mortality, and incidence of hypoglycaemia, nasopharyngitis, urinary tract infection, upper respiratory infection, nausea, vomiting, and diarrhoea., Results: 27 reports of 19 studies including 7136 patients randomised to a DPP-4 inhibitor and 6745 patients randomised to another hypoglycaemic drug were eligible for the systematic review and meta-analysis. Overall risk of bias for the primary outcome was low in three reports, unclear in nine, and high in 14. Compared with metformin as monotherapy, DPP-4 inhibitors were associated with a smaller decline in HbA(1c) (weighted mean difference 0.20, 95% confidence interval 0.08 to 0.32) and in body weight (1.5, 0.9 to 2.11). As a second line treatment, DPP-4 inhibitors were inferior to GLP-1 agonists (0.49, 0.31 to 0.67) and similar to pioglitazone (0.09, -0.07 to 0.24) in reducing HbA(1c) and had no advantage over sulfonylureas in the attainment of the HbA(1c) goal (risk ratio in favour of sulfonylureas 1.06, 0.98 to 1.14). DPP-4 inhibitors had a favourable weight profile compared with sulfonylureas (weighted mean difference -1.92, -2.34 to -1.49) or pioglitazone (-2.96, -4.13 to -1.78), but not compared with GLP-1 agonists (1.56, 0.94 to 2.18). Only a minimal number of hypoglycaemias were observed in any treatment arm in trials comparing a DPP-4 inhibitor with metformin as monotherapy or with pioglitazone or a GLP-1 agonist as second line treatment. In most trials comparing a DPP-4 inhibitor with sulfonylureas combined with metformin, the risk for hypoglycaemia was higher in the group treated with a sulfonylurea. Incidence of any serious adverse event was lower with DPP-4 inhibitors than with pioglitazone. Incidence of nausea, diarrhoea, and vomiting was higher in patients receiving metformin or a GLP-1 agonist than in those receiving a DPP-4 inhibitor. Risk for nasopharyngitis, upper respiratory tract infection, or urinary tract infection did not differ between DPP-4 inhibitors and any of the active comparators., Conclusion: In patients with type 2 diabetes who do not achieve the glycaemic targets with metformin alone, DPP-4 inhibitors can lower HbA(1c), in a similar way to sulfonylureas or pioglitazone, with neutral effects on body weight. Increased unit cost, which largely exceeds that of the older drugs, and uncertainty about their long term safety, however, should also be considered.

Twenty-four single-nucleotide polymorphisms (SNPs) have been reproducibly associated with obesity. We performed a follow-up study for obesity in the Greek adult population. A total of 510 obese and 469 lean adults were genotyped for 24 SNPs. We tested the association with obesity status using logistic regression and we evaluated the combined genetic risk of 24 SNPs by calculating the area under the receiver-operating characteristic (ROC) curves. We nominally replicated the association with obesity (BMI ≥30 kg/m(2)) of six SNPs in or near the FTO, MC4R, TMEM18, PRL, AIF1, and PCSK1 loci (1.28 ≤ odds ratio (OR) ≤ 1.35; 0.004 ≤ P ≤ 0.043). The discrimination ability for obesity was slightly stronger (P = 9.59 × 10(-6)) when the genetic information of the 24 SNPs was added to nongenetic risk factors (area under the curve (AUC) = 0.722) in comparison with nongenetic factors analyzed alone (AUC = 0.685). Our data suggest that SNPs in or near the FTO, MC4R, TMEM18, PRL, AIF1, and PCSK1 loci contribute to obesity risk in the Greek population.

Aim: To assess the reduction in estimated cardiovascular disease (e-CVD) risk after multifactorial treatment for 6 months and follow this change during the next 3-years., Patients-Methods: This prospective, randomized, target driven study included 1,123 subjects (512/611 men/women, aged 45-65 years) with metabolic syndrome (MetS) without diabetes or CVD referred to specialist outpatient clinics. Patients were randomized to two treatment groups: group A with low density lipoprotein cholesterol (LDL-C) target of < 100 mg/dl and group B with a target of < 130 mg/dl. Atorvastatin was used in both groups on top of optimal multifactorial treatment, (quinapril, amlodipine, hydrochlorothiazide for hypertension, metformin for impaired fasting glucose, and orlistat for obesity). The e-CVD risk was calculated using the Framingham, the PROCAM and Reynold's equations., Results: Reductions in e-CVD risk at 6 months were > 50%in all patients, but were superior in group A and in women. Reductions were even greater during the next 3-years and were mainly attributed to changes in lipid profile. Actual CVD events were 1 in group A and 13 in group B; p=0.0012., Conclusions: Attaining the treatment target of LDL-C < 100 mg/dl within multifactorial treatment of MetS by expert clinics, is achievable and beneficial even in patients without diabetes or known CVD. This induces a considerable e-CVD risk reduction in MetS patients. Actual CVD events were negligible, suggesting that e-CVD risk overestimates actual CVD risk in MetS, at least in patients achieving LDL-C < 100 mg/dl [ClinicalTrials.gov ID: NCT00416741].

Aim: To assess the effects of long-term multifactorial intervention on renal function and serum uric acid (SUA) levels and their association with estimated cardiovascular disease (eCVD) risk and actual CVD events., Methods: This prospective, randomized, target-driven study included 1123 subjects (45.6% men, age 45-65 years) with metabolic syndrome (MetS) but without diabetes or CVD. Patients were randomized to multifactorial treatment. Atorvastatin was titrated from 10-80 mg/day aiming at a low density lipoprotein cholesterol (LDL-C) target of <100 mg/dl (group A) or an LDL-C target of <130 mg/dl (group B). Changes in estimated glomerular filtration rate (eGFR) and SUA levels were recorded in all patients and in the subgroup with stage 3 chronic kidney disease (CKD; eGFR = 30-59 ml/min/1.73 m(2); n = 349). We used ANOVA to compare changes within the same group, unpaired Student t-test to compare results between groups at specific time points, and log-rank test to compare event free survival., Results: The eCVD-risk reduction was greater in group A. In the overall study population, eGFR increased by 3.5% (p < 0.001) and SUA levels fell by 5.6% (p < 0.001). In patients from group A with stage 3 CKD (group A1; n = 172), eGFR increased by 11.1% (p < 0.001) from baseline and by 7.5% (p < 0.001) in group B1 (n = 177; p < 0.001 vs. the change in group A1). The corresponding fall in SUA levels was 10.7% in group A1 (p < 0.001 vs. baseline) and 8.3% in group B1 (p < 0.001 vs. baseline and group A1). These changes were mainly attributed to atorvastatin treatment. Among the CKD stage 3 patients there were no CVD events in group A1, while 6 events occurred in group B1 (p = 0.014)., Conclusions: Multifactorial intervention in patients with MetS without established CVD improved renal function and reduced SUA levels. These changes were more prominent in stage 3 CKD patients and might have contributed to the reduction in eCVD risk and clinical events. Original study registration number [ClinicalTrials.gov ID: NCT00416741].

Objective: To estimate the accuracy of Neuropad for the diagnosis and staging of distal symmetric polyneuropathy (DPN) across different stages of neuropathy, using multiple-level likelihood ratios (LRs) to interpret the time necessary to complete the color change of the test., Research Design and Methods: We conducted a cross-sectional, cohort-type diagnostic accuracy study in 251 consecutive adult type 2 diabetic patients with no peripheral arterial disease or other potential causes of neuropathy, who were recruited between January 2005 and December 2008 from the diabetes outpatient clinics in Alexandroupolis Hospital, Greece. Patients were tested for DPN by means of the neuropathy disability score (NDS) and Neuropad. Multiple-level LRs for time to complete color change were calculated across different stages of neuropathy., Results: The areas under the curve for the diagnosis of any (NDS of ≥3), at least moderate (NDS of ≥6), or severe (NDS of ≥9) DPN were 0.91, 0.96, and 0.97, respectively. The calculation of multiple-level LRs showed that time to complete color change <360 s suggested the absence of neuropathy. Values between 360 and 1,000 s were indicative of mild neuropathy. Finally, values between 1,000 and 1,200 or >1,200 s were strongly suggestive of moderate or severe DPN, respectively., Conclusions: Neuropad could be used as a triage test for the diagnosis and staging of DPN in patients with type 2 diabetes, prompting referral to specialized care setting.

In the present study the effect of high glucose concentrations, insulin, PPARγ activators (rosiglitazone) and NHE-1 inhibitors (cariporide) in atherosclerosis-related functions of human monocytes was investigated. Monocyte adhesion to laminin-1, collagen type IV and endothelial cells, as well as monocyte migration through the same substrates were studied. Incubation of the monocyte suspension with high glucose concentrations, insulin and rosiglitazone induced all the studied atherosclerosis-related functions of the monocytes. In all these functions the addition of cariporide counteracted the activity of glucose, insulin and rosiglitazone. The use of antigen for β1 integrin also counteracted the activity of the above in monocyte adhesion in all three substrates. The data of the present study suggests that PPARγ activation in monocytes induces atherosclerosis, and that NHE-1 and β1 integrin play an important role in the beginning of atherosclerosis.

We assessed the effect of glucose and insulin on human monocytes. Monocytes were isolated from 16 healthy obese and 10 lean healthy participants. Insulin sensitivity was assessed by euglycemic hyperinsulinemic clamp. Obese participants were subdivided into 2 subgroups: insulin sensitive (IS) and insulin resistant (IR). Monocyte oxidized low-density lipoprotein (oxLDL) phagocytosis was assessed pre and poststimulation in vitro with glucose or insulin. Experiments were repeated after incubation with a Na(+)/H(+) exchanger-1 inhibitor ([NHE-1]; cariporide) or rosiglitazone. Glucose increased oxLDL phagocytosis in all groups studied (at 1 or 3 hours incubation; P = .037-.002). Insulin increased oxLDL phagocytosis in all groups studied after 1-hour incubation (P = .027-.015) but not at 3 hours. Incubation with cariporide attenuated oxLDL phagocytosis except in the obese IS group. Rosiglitazone eliminated glucose- and insulin-induced increase in oxLDL phagocytosis in all studied groups. Glucose and insulin induce oxLDL phagocytosis.

We assessed the effect of epinephrine on human monocytes. Monocytes were isolated from 16 healthy obese and 10 lean healthy subjects. Insulin sensitivity was assessed by euglycemic hyperinsulinemic clamp. Obese subjects were subdivided into 2 sub-groups, insulin sensitive (IS) and insulin resistant (IR). Monocyte properties [attachment to laminin 1, migration through laminin 1, oxidized-low density lipoprotein (oxLDL) phagocytosis] were assessed pre- and post-stimulation in vitro with epinephrine. Experiments were repeated after incubation with a Na(+)/H( +) exchanger-1 inhibitor (NHE-1) (cariporide). Epinephrine increased monocyte attachment to laminin in lean and obese IR subjects through involvement of NHE-1, PKC, NO synthase, NADPH oxidase and actin polymerization. In contrast, epinephrine did not affect monocyte migration. Epinephrine increased oxLDL phagocytosis in all groups studied. Incubation with cariporide attenuated oxLDL phagocytosis. Epinephrine induces monocyte dysfunction which may be atherogenic.

Importance of the Field: The sodium/hydrogen exchanger-1 (NHE-1/SLC9A1) is a ubiquitous plasma membrane protein whose main role is maintenance of intracellular pH and volume. NHE-1 plays a role in atherogenesis; however, its clinical relevance has not yet been established., Areas Covered in This Review: We herein review the contribution of NHE-1 in atherogenesis (namely its effect on endothelial cells, monocytes, smooth muscle cells and platelets)., What the Reader Will Gain: Studies have shown that NHE is involved in atherogenesis-related properties of isolated monocytes. We also consider the relationship between NHE-1 and vascular risk factors such as obesity, diabetes mellitus, hypertension, dyslipidemia and inflammation., Take Home Message: Even though clinical trials with certain NHE-1 inhibitors have had discouraging results, NHE-1 cannot be excluded as a potential future therapeutic target for the prevention and/or treatment of atherosclerosis.

We assessed the effect of leptin on human monocytes. Monocytes were isolated from 16 healthy obese and 10 lean healthy participants. Insulin sensitivity was assessed by euglycemic hyperinsulinemic clamp. Obese participants were subdivided into 2 subgroups, insulin sensitive (IS) and insulin resistant (IR). Monocyte properties (attachment to laminin 1, migration through laminin 1, surface expression of CD36, oxidized low-density lipoprotein [oxLDL] phagocytosis) were assessed pre- and poststimulation in vitro with leptin. Experiments were repeated after incubation with rosiglitazone and a Na(+)/H(+) exchanger-1 inhibitor (cariporide). We found a significant correlation between insulin resistance and monocyte attachment to laminin and oxLDL phagocytosis. Leptin increased the atherosclerosis-related properties of monocytes in all groups, apart from surface expression of CD36 in IS obese participants. Incubation with rosiglitazone or cariporide attenuated the observed effects. Leptin induces monocyte dysfunction which may be atherogenic. Correlation with insulin resistance suggests that atherosclerosis might be accelerated before the onset of diabetes.

Background: Sodium/hydrogen exchanger-1 (NHE-1) contributes to maintaining intracellular pH (pHi). We assessed the effect of glucose, insulin, leptin and adrenaline on NHE-1 activity in human monocytes in vitro. These cells play a role in atherogenesis and disturbances in the hormones evaluated are associated with obesity and diabetes., Methods and Results: Monocytes were isolated from 16 healthy obese and 10 lean healthy subjects. NHE-1 activity was estimated by measuring pHi with a fluorescent dye. pHi was assessed pre- and post-incubation with glucose, insulin, leptin and adrenaline. Experiments were repeated after adding a NHE-1 inhibitor (cariporide) or an inhibitor of protein kinase C (PKC), nitric oxide synthase (NOS), nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, phosphoinositide 3-kinases (PI3K) or actin polymerization. Within the whole study population, glucose enhanced NHE-1 activity by a processes involving PKC, NOS, PI3K and actin polymerization (p = 0.0006 to 0.01). Insulin-mediated activation of NHE-1 (p = <0.0001 to 0.02) required the classical isoforms of PKC, NOS, NADPH oxidase and PI3K. Leptin increased NHE-1 activity (p = 0.0004 to 0.04) through the involvement of PKC and actin polymerization. Adrenaline activated NHE-1 (p = <0.0001 to 0.01) by a process involving the classical isoforms of PKC, NOS and actin polymerization. There were also some differences in responses when lean and obese subjects were compared. Incubation with cariporide attenuated the observed increase in NHE-1 activity., Conclusions: Selective inhibition of NHE-1 in monocytes could become a target for drug action in atherosclerotic vascular disease.

Gestational diabetes mellitus (GDM) is defined as any degree of glucose intolerance with onset or first recognition during pregnancy. Women with GDM and their offspring have an increased risk of developing type 2 diabetes mellitus in the future. The global incidence of GDM is difficult to estimate, due to lack of uniform diagnostic criteria. Various diagnostic criteria have been proposed. The benefit of treating GDM has also been controversial. The clinical significance of treating maternal hyperglycemia was made evident in the Hyperglycemia and Adverse Pregnancy Outcomes (HAPO) study. The HAPO study demonstrated that there is a continuous association of maternal glucose levels with adverse pregnancy outcomes and served as the basis for a new set of diagnostic criteria, proposed in 2010 by the International Association of Diabetes and Pregnancy Groups (IADPSG). According to these criteria the diagnosis of GDM is made if there is at least one abnormal value (≥92, 180 and 153 mg/dl for fasting, one-hour and two-hour plasma glucose concentration respectively), after a 75 g oral glucose tolerance test (OGTT).

Unlabelled: The aim of this study was to examine the potential correlation of WHO-5 well-being index with glycaemic control and chronic complications in subjects with type 2 diabetes. The study included 156 subjects (73 men, mean age 64.05+/-9.11 years, mean diabetes duration 12.22+/-5.61 years). Well-being was assessed by the WHO-5 score via a validated questionnaire comprising 5 questions (Q1-Q5). HbA (1c) showed a significant negative correlation with overall WHO-5 score (r (s)=-0.248, p=0.002) and individual Q1-Q4 scores (r (s)=-0.262, p=0.001; r (s)=-0.248, p=0.002; r (s)=-0.207, p=0.009 and r (s)=-0.169, p=0.035 respectively). Subjects with adequate glycaemic control (HbA (1c) < 7%, n=67) had a significantly higher WHO-5 score in comparison to those with inadequate glycaemic control (HbA (1c) >or= 7%, n=89) (mean+/-SD: 19.69+/-5.47 vs. mean+/-SD: 17.11+/-6.38, p=0.011). Finally, WHO-5 score was significantly (p=0.013) lower in subjects with neuropathic pain than in those without neuropathic pain., Conclusions: In type 2 diabetic subjects, glycaemic control shows a significant correlation with well-being, while neuropathic pain is associated with lower well-being score., ((c) J. A. Barth Verlag in Georg Thieme Verlag KG Stuttgart, New York.)

Background: Obesity is a rapidly expanding epidemic in Western societies, with rates of more than 30% across Europe, and it is associated with an increased risk of metabolic disturbances. Previous reports have documented an association of reduced physical activity and abstinence from the traditional Mediterranean diet (MD) with increased mortality rate and prevalence of obesity in a population of Greek subjects., Objective: The aim of the present study was to evaluate and analyze the dietary habits in a population of Greek overweight and obese subjects and to investigate the potential associations between those patterns and the prevalence of metabolic syndrome components., Methods: The study recruited 226 consecutive adult (30 men, 169 women) overweight or obese (body mass index >25 kg/m(2)) individuals attending the Metabolic Diseases Unit. Medical history, dietary history, and anthropometric parameters were recorded during the first visit. Fasting blood samples were collected for biochemistry assaying., Results: According to the nutrient intake history and Mediterranean Diet Scale (MDS), participants were divided into 3 groups: those adhering to the MD and those not following the MD, who were further subdivided into the high-carbohydrate (HC) and high-fat (HF) diet groups according to the source of maximum energy intake. Adherence to the MD was associated with a lower prevalence of metabolic syndrome (27.3%, 69.2%, and 60.4% in MD, HC, and HF respectively, p = 0.006), lower low-density lipoprotein cholesterol (p = 0.009, MD vs. HF), and lower postchallenge glucose values (p = 0.028, MD vs. HF)., Conclusions: Adherence to the MD seems to be declining among Greek overweight and obese subjects, a phenomenon that is associated with an increase in the prevalence of the metabolic syndrome.

Leptin, a 16-kDa cytokine produced mainly by the adipose tissue, is known to increase energy expenditure while at the same time lowering food intake by acting directly on the hypothalamus. ObRb, the leptin receptor mostly involved in intracellular signaling, is expressed in a wide range of tissues, thus allowing leptin to affect a much broader diversity of biological processes. High concentrations of leptin are encountered in patients with hyperleptinemia, a condition which very often accompanies obesity and which is a direct result of leptin resistance. In the present study, moderate and high concentrations of leptin (16 and 160 ng/ml) were mostly utilized in order to investigate the role of this cytokine in oxidative stress levels in human monocytes. Leptin was found to increase oxidative species production as measured with 2',7'-dichlorodihydrofluorescein diacetate (general marker of oxidative species, but not O2-*) and dihydroethidium (marker of O2-*). Surprisingly, it also augmented superoxide dismutase activity. Inhibition of the Na+-H+ exchanger isoform 1 (NHE1) also inhibited leptin-induced superoxide anion production but at the same time amplified leptin-induced production of other oxidative species. Signaling proteins such as phosphoinositide 3 kinase and conventional isoforms of protein kinase C (alpha-, beta(i)-, beta(ii)-), as well as NADPH oxidase, also participated in leptin signaling. Finally, leptin was found to increase glutathionylation levels of NHE1-bound heat shock protein 70 kDa (Hsp70) but not Hsp70 binding to NHE1.

Monocyte-extracellular matrix interactions have been implicated in atherosclerosis pathophysiology. Laminin, the main basement membrane protein contains cell binding domains that can be cryptic, presented only after protein modification. In the present study we evaluated monocyte attachment to laminin-1 in the presence of ATP. Monocytes were derived from either healthy volunteers or patients with diabetes mellitus type II. For the estimation of monocyte attachment to laminin the myeloperoxidase assay was used. Monocytes derived from diabetic patients, showed an increased ability to attach to laminin (p = 0.0055). The presence of ATP increased the attachment of control monocytes to laminin (p = 0.0022). On the contrary, the presence of ATP did not affect the attachment of monocytes derived from diabetic patients to laminin. Our results indicate a modified interaction between monocytes and laminin-1 in diabetes mellitus.

Aim: To substantially increase awareness, treatment and effective control of the metabolic syndrome (MetS) and its components., Subjects and Methods: This is a pilot best practice implementation enhancement programme to reduce the estimated cardiovascular disease (CVD) risk in 628 MetS patients with or without diabetes or CVD by improving quality of care. A baseline visit was followed by action to improve adherence to lifestyle advice and drug treatment for CVD risk factors by physicians specifically trained to implement guidelines. Finally, after 6 months, a single-page form was completed, showing if patients were at CVD risk factor target. If not, there was an analysis of the reason why., Results: The programme was effective in improving utilization of evidence-based treatment in 628 MetS patients. There was a substantially greater patient perception of MetS, an enhancement in compliance with lifestyle advice and increased prescription of evidence-based medication, leading to a 48% (p < 0.0001) improvement in estimated CVD risk. There was a substantial increase in the number of subjects on target for specific CVD risk factors., Conclusions: This is the first study to increase adherence to multiple interventions for all MetS components on an outpatient basis, in both primary care and teaching hospital settings. Physician and patient education, distribution of printed guidelines and brochures, and completion of a single-page form motivated both physicians and patients to achieve multiple CVD risk factor guideline goals. The absence of a control group is a limitation of this study. Further work is also needed to establish if the improvements observed are sustained on a long-term basis.

Background/aims: Leptin, a 16-kDa cytokine that is released mainly by the adipose tissue, is known to affect a wide assortment of processes, ranging from energy homeostasis to angiogenesis and the immune response. In the present study, the effect of leptin on atherosclerosis-related properties of human monocytes was investigated. methods: Monocytes were isolated from whole blood obtained from healthy donors who had normal body mass index values. Pharmacological inhibition of specific signaling proteins was implemented. Fluorescence spectrometry and immunofluorescence techniques, as well as ELISA methods, were utilized. Leptin dose response curves were determined for each type of experiment., Results: Leptin (160 ng/ml) was found to augment monocyte adhesion to laminin-1 and its migration through this glycoprotein, which is one of the main components of the extracellular matrix. Additionally, leptin increased CD36-receptor surface expression, as well as moderately oxidized low-density lipoprotein (oxLDL(3)) uptake levels., Conclusion: Leptin amplifies the pro-atheromatic properties of human monocytes through a complex signaling net which involves the Na(+)/H(+) exchanger isoform-1, the actin cytoskeleton, phosphoinositide 3-kinase, certain conventional isoforms of protein kinase C and NADPH oxidase., (Copyright (c) 2008 S. Karger AG, Basel.)

Non-alcoholic fatty liver disease (NAFLD) is a clinicopathologic entity increasingly recognized as a major health burden in developed countries. It includes a spectrum of liver damage ranging from simple steatosis to nonalcoholic steatohepatitis (NASH), advanced fibrosis, and rarely, progression to cirrhosis. Recent studies emphasize the role of insulin resistance, oxidative stress and subsequent lipid peroxidation, proinflammatory cytokines, adipokines and mitochondrial dysfunction in the development and progression of NAFLD. Furthermore, accumulating evidence supports an association between NAFLD and metabolic syndrome. Although the data are mainly epidemiological, the pathogenesis of NAFLD and metabolic syndrome seems to have common pathophysiological mechanisms, with focus on insulin resistance as a key factor. This review summarizes the current knowledge on the epidemiology, pathophysiology and diagnosis of both NAFLD and metabolic syndrome and the findings that strongly support the association of nonalcoholic fatty liver disease as a possible component in the cluster of metabolic syndrome.

Objective: To investigate the relationship between MKKS gene variations, obesity-related traits and features of the metabolic syndrome (MS) in the Greek population., Design and Subjects: Genotype and haplotype analysis was carried out for six known MKKS gene polymorphisms (534C>T, 985+16T>G, 985+33C>G, 986-29A>T, 1161+58A>G and 1595G>T) in 220 obese subjects (body mass index > or =30 kg/m(2)) and 330 non-obese controls., Results: Genotype frequencies of the 985+16T>G, 986-29A>T and 1595G>T SNPs were significantly different between obese and non-obese individuals (P=0.0016, 0.0196 and 0.0069, respectively). Obese carriers of the risk alleles of the above three polymorphisms had a significantly increased prevalence of arterial hypertension. Furthermore, obese carriers of the G allele for the 985+16T>G polymorphism had an increased prevalence of type 2 diabetes mellitus and of MS component traits. A new polymorphism was detected, namely a C to T substitution at position 1129 (1129C>T or N377N). Frequency of the T allele for the 1129C>T polymorphism was significantly higher in control individuals than in obese subjects (P=0.0253). Haplotype TGTGT was more prevalent in obese than in controls (P=0.0002) and was associated with increased prevalence of the MS in obese subjects (P<0.0001)., Conclusion: Our results suggest that genetic variation in the MKKS gene may play a role in the development of obesity and the metabolic syndrome.

The interactions between monocytes and extracellular matrix proteins have been implicated in atherosclerosis pathophysiology. In the present study we evaluated monocyte attachment and migration through oxidized and non-oxidized collagen IV. Monocyte attachment was tested on microwells coated with either native or oxidized collagen IV. Monocyte migration through collagen IV was examined on transwells. Monocytes derived from patients with diabetes mellitus showed an increased ability to attach and migrate through collagen IV as compared to those derived from healthy volunteers. Moreover, control monocytes attached to oxidized collagen at a higher degree, while they migrated through oxidized collagen at a lower degree, as compared to the native protein. Our results also showed the involvement of the alpha2 integrin subunit in the above phenomena suggesting a modified interaction between monocytes and collagen IV in diabetes mellitus.

Angiotensin II, a potent vasoconstrictor, has been demonstrated to be involved in the pathogenesis of atherosclerosis, and its complications. Na(+)/H(+) exchanger isoform-1 (NHE1) is related to hypertension activation and can augment atherosclerosis-related functions in human monocytes. The purpose of this study is to investigate in monocytes the signal transduction pathway induced by angiotensin II, in which the Na(+)/H(+) exchanger (NHE1) takes part. Monocytes were isolated, and intracellular pH (pHi) was measured by the use of Bis-(carboxyethyl)-5(6)-carboxy-fluorescein acetoxymethylester. Superoxide anions were measured by nitroblue tetrazolium. Monocyte binding to laminin-1 was quantified using the myeloperoxidase assay. Angiotensin II caused a significant increase in pHi of monocytes, which indicates NHE activation. Cariporide, an NHE1 inhibitor, GF109203X, and Gö6976, inhibitors of isoforms of protein kinase C (PKC), diphenyleneiodonium chloride, the inhibitor of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, and N(omega)-Nitro-L-arginine methyl ester hydrochloride, the inhibitor of nitric oxide (NO) synthase, reversed this effect. Moreover, it was shown that angiotensin II caused an increase in superoxide ion (O(2)(-.)) levels as well as an increase in monocytes' adhesion to laminin-1, in relation to controls. The use of cariporide inhibited these effects. Furthermore, angiotensin II caused an increase in pHi, which was reversed by cariporide in monocytes derived from hypertensive patients. Consequently, in human monocytes angiotensin II caused NHE1 activation through pathways involving isoforms of PKC with the participation of O(2)(-.) and NO. In addition, a link between angiotensin II and the atherogenic properties of monocytes was shown, where NHE1 plays a central role.

Objectives: Determination of the prooxidant-antioxidant balance (PAB) in patients with angiographically defined coronary artery disease (CAD+) by a modified PAB assay and presentation of PAB value as a novel cardiovascular risk factor., Design and Methods: For 61 patients with CAD+ and 63 healthy volunteers, the PAB were measured and its correlation was determined with anthropological and clinical parameters., Results: A significant increase of the PAB value was observed in patients in comparison to control group. A correlation, which is not quite significant, was noted between angiographic finding (number of diseased vessel) and the PAB values in patients. A significant positive correlation was established between the PAB value and systolic blood pressure, diastolic blood pressure, smoking, fasting blood sugar and serum urea concentration; and a significant negative correlation was established between PAB value and serum creatinine and bilirubin., Conclusions: This study shows that the PAB value may be considered as a cardiovascular risk factor. Further clinical research is needed to substantiate the potency of the PAB value as a cardiovascular risk factor.

The activation of sodium/hydrogen exchanger (NHE) is associated with a variety of cell functions like cell adhesion, migration, proliferation, and apoptosis. Since its discovery, 9 NHE isoforms have been identified, but the most widely spread and the most important for the cellular functions is NHE-1. This ubiquitously expressed sodium/hydrogen exchanger (NHE-1) plays a central housekeeping role in all cells regulating cell volume and internal pH (pHi). At physiological pHi, NHE-1 is essentially inactive but it is extremely sensitive to pHi changes, being rapidly activated by small intracellular hydrogen concentration increases. NHE-1 activity can be stimulated via a series of cell surface receptors, including tyrosine kinase, G-protein-coupled, and integrin receptors. These signals converge, regulating the affinity of the internal hydrogen-binding site. NHE-1 also is a plasma membrane-anchoring protein for the cytoskeleton. Cytoskeleton anchoring of NHE-1 is important for cell adhesion to extracellular matrix proteins and cell migration. Moreover, NHE-1 plays the role of a "scaffold" for the building of various intracellular signaling molecule clusters.

Drugs are known to be a cause of pulmonary eosinophilia and several case reports of acute eosinophilic pneumonia associated with the use of cocaine have been reported. The changing pattern of heroin use, with a shift from intravenous use to smoking/inhalation of the substance, may lead to increased prevalence of heroin-induced pulmonary eosinophilia. We report on a case of a patient who had been inhaling heroin for about ten years. He presented with fever, cough, dyspnea and pleuritic chest pain. Chest radiograph showed unilateral pleural effusion with segmental atelectasis. Examination of pleuritic fluid aspirate and bronchoalveolar lavage fluid revealed significant eosinophilia. He was diagnosed with acute eosinophilic pneumonia. Rapid remission was achieved after heroin abstinence and initiation of corticosteroid treatment.

Objective: To assess insulin sensitivity in young adult normoglycemic beta-thalassaemia major patients., Methods: We measured insulin sensitivity with the euglycemic insulin clamp in 10 young adult (mean age 24.85 +/- 2.45 yrs) normoglycemic beta-thalassaemia major patients and 10 sex- & age-matched controls. Liver iron accumulation was assessed by magnetic resonance imaging (MRI)., Results: Glucose infusion rate (M) required to maintain euglycemia was significantly reduced in thalassaemic patients compared to controls (261.5 +/- 63.5 mg/m2 x min vs. 355.6 +/- 35.3 mg/m2 x min, P = 0.008). Consequently, significantly reduced in the thalassaemic group were also tissue sensitivity to insulin (M/I(s-s)) and glucose metabolic clearance rate (M/G(s-s)). There was significant negative correlation between ferritin levels and glucose infusion rate (r = -0.918 P = 0.004). No significant correlations were observed between age, body mass index, daily transfusional iron accumulation, liver iron content and any of the euglycemic clamp parameters. Fasting insulin levels were significantly increased in patients with beta-thalassaemia major compared to controls (P = 0.01), and had significant negative correlation to MRI-derived liver iron content (r = -0.733, P = 0.03)., Conclusions: Our data indicate that reduced insulin sensitivity resulting in hyperinsulinaemia precedes the manifestation of glucose intolerance in patients with beta-thalassaemia major. Insulin resistance seems to correlate with increased serum ferritin levels.

Objectives: Monocyte-extracellular matrix interactions have been implicated in atherosclerosis pathophysiology. In the present study we evaluated the oxidation of laminin by monocytes derived from either diabetic patients or healthy volunteers. Moreover, reactive oxygen species production was measured. Monocyte attachment and migration through oxidized and non-oxidized laminin were also studied., Design and Methods: Laminin oxidation was tested by a sensitive ELISA assay in isolated monocytes. ROS production was measured with fluorescent indicators. 35S-methionine was used for evaluating monocyte attachment. Monocyte migration through laminin was examined on transwells., Results: Monocytes derived from patients with diabetes mellitus showed an increased ability to carbonylate and attach to laminin. Diabetic monocytes produced increased levels of ROS as compared to controls. Our results showed the involvement of the alpha2 integrin subunit in monocyte attachment to both native and oxidized laminin in control and diabetic monocytes., Conclusions: The results indicate a modified interaction between monocytes and laminin in diabetes.

Objectives: The application of a novel assay for the direct measurement of prooxidant-antioxidant balance (PAB) in type II diabetes and the evaluation of antioxidant therapy., Design and Methods: The assay is based on 3,3',5,5'-tetramethylbenzidine and its cation, used as a redox indicator participating in two simultaneous reactions. PAB was determined in the sera of healthy volunteers and type II diabetes patients. The results were compared with clinical and biological parameters, protein oxidation markers, as well as the results of antioxidant and prooxidant assays. PAB, after administration of vitamins C and E for 1 day, 1 month and 2 months was also determined., Results: Increased PAB was found in the patients' group and correlated with parameters involved in diabetic complications, protein oxidation markers, antioxidant and prooxidant assays. One day after vitamin administration, a significant shift of PAB towards antioxidants was observed. PAB remained unchanged after 1 month and changed marginally in favor of prooxidants in the second month of the therapy., Conclusions: These results indicate that the measurement of PAB may be useful to identify and follow-up patients who need antioxidant therapy.

The objective of this article is to investigate the influence of endothelin-1 (ET-1) on human monocyte Na(+)/H(+) exchanger (NHE) activity and on the atherosclerosis-related monocyte functions. ET-1 caused an increase in pHi and in (22)Na influx of monocytes. A reversal of ET-1 effect on pHi was observed in the presence of the NHE1 inhibitor, cariporide. In addition, the activation of NHE1 by ET-1 was mediated via protein kinase C (PKC), mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase (PI3K), and NADPH oxidase. Also, a link between ET-1 and nitric oxide (NO) was observed. Furthermore, after ET-1 treatment, an increase of the adhesive capacity, the migration ability on laminin and CD36 expression of monocytes, was observed; using cariporide this increase was abolished. Our results showed that ET-1 induces a signaling pathway with the involvement of PKC, MAPK, PI3K, and NADPH oxidase where NHE1 plays a key role. ET-1 also plays a significant role in atherosclerosis-related functions of human monocytes, via NHE1 activation.

The oxidative modification of proteins has been shown to play a major role in a number of pathological processes. One such modification is the addition of the carbonyl groups to the amino acid residue in proteins. For the measurement of the carbonyl groups in low concentration protein samples, we have modified the ELISA (enzyme-linked immunosorbent assay) method that was developed by Buss et al. [Buss, I. H; Chan, T. P.; Sluis, K. B.; Domigan, N. M.; Winterbourn, C. C. Protein carbonyl measurement by a sensitive ELISA method. Free Radic. Biol. Med.23:361-366; 1997 ]. In the modified method, protein samples diluted in phosphate-buffered saline were adsorbed to wells of an ELISA plate and then reacted with dinitrophenylhydrazine (DNPH). The protein-conjugated DNPH was probed by a commercial anti-DNPH antibody, and then a second antibody conjugated with horseradish peroxidase was added for quantification. The method was calibrated using oxidized albumin, and required only 5 mug protein. This obviated the need to concentrate protein in experimental and clinical samples with low amounts of protein. In addition the effect of TCA on carbonyl measurement is eliminated. The standard curve was linear in the range of 0-3.36 nmol carbonyls/mg protein, which is the range within which clinical samples fell. The results correlated well with the colorimetric carbonyl assay. The method was used to analyze the amount of protein carbonyl in aqueous humor and diluted plasma samples.