Multicenter, open-label, non-randomized, non-interventional observational study of safety of treatment initiation with a biphasic insulin aspart.

BACKGROUND: Type 2 diabetes mellitus is characterized by hyperglycaemia. Incidence and progression of microvascular complications have been shown to undergo reduction with lowered glucose levels. METHODS: This is an open-label, non-randomized, non-interventional, observational study of safety and efficacy of BiAsp 30, enrolling 2223 patients with type 2 diabetes mellitus. Patients previously treated with oral antidiabetic drugs and/or human insulin were recruited, provided they had been hospitalized for at least 3 days. Data were collected during a single multi-stage visit lasting up to 6 days, during the patient's hospitalization period. A mild hypoglycaemic episode was defined as blood glucose measurement < 56 mg/dL (3.1 mmol/L). A severe hypoglycaemia was hypoglycaemic episode requiring assistance of another person. RESULTS: The total number of hypoglycaemic episodes decreased over time of follow-up. The intensity of severe hypoglycaemic episodes decreased by almost a third from day to day (IRR = 0.64, 95%, p < 0.001). However, intensity of mild hypoglycaemic episodes was the same over the subsequent days of follow-up (IRR = 0.97, p > 0.1). Mean blood glucose concentration decreased from 202.0 mg/dL to 157.8 mg/dL after 3 days and to 138.3 mg/dL after 6 days (p < 0.001). CONCLUSION: This study highlights the safety, efficacy and ease of initiation of insulin BiAsp 30 therapy for treatment of type 2 diabetes mellitus.

The influence of incretin mimetics on cardiovascular risk factors in diabetes.

The authors discuss the strategy of use of incretin hormones in type 2 diabetes treatment in the context of cardiovascular complications. The results of the phase III study on human GLP-1 (Glucagon-like peptide-1) analogue-liraglutide have been presented under common acronym LEAD (Liraglutide-Effect and Action In Diabetes). The liraglutide therapy improved glycemic control with low hypoglycemia risk and decreased glycated hemoglobin by an average 1,13%. Decreases in systolic pressure and significant body weight loss were observed. Not only did the index describing beta cells function HOMA-B improve but also did the ratio of insulin to proinsulin. Summing up, incretin hormones beneficially influence blood glucose level, moreover, their use decreases blood pressure and body weight which might indicate their positive influence on cardiovascular system in diabetic patients.

Anti-TNF rescue CD4+Foxp3+ regulatory T cells in patients with type 1 diabetes from effects mediated by TNF.

The presence of low-grade chronic inflammation is a known feature of long standing diabetes type 1. The association between serum level of several markers of inflammation and severity of DM1 was proven. Serum concentrations of TNF were reported to be elevated in diabetic patients, especially those who developed diabetic complications. Lately, it has been also shown that TNF may impair the subset of naturally arising regulatory T cells, which control autoimmunity. The presented study, for the first time, shows the regulatory T cells in the context of an inflammatory environment that is present in patients with type 1 diabetes. It indicates that TNF reduces the number and frequency of regulatory CD4(+)Foxp3(+) T cells in children with diabetes type 1 and that in vitro treatment with anti-TNF antibody seems to rescue this cell subset from its defective effects.

[Regulatory T lymphocytes expressing L-selectin in children and adolescents with type 1 diabetes mellitus]. / Limfocyty T regulatorowe z ekspresja L-selektyny u dzieci i mlodziezy z przewlekla cukrzyca typu 1.

INTRODUCTION: Quantitative and/or qualitative dysfunctions in a subset of naturally arising regulatory T lymphocytes may have impact on autoimmune disease development, including diabetes type 1. CD62L is a homing receptor that directs T lymphocytes to lymph nodes. Studies conducted on NOD mice showed that depletion of Tregs expressing CD62L results in diabetes and only CD62Lhigh Tregs are able to protect against the disease. AIM OF THE STUDY: The purpose of the paper was to analyze the regulatory T lymphocyte subset with CD62L expression in children with diabetes type 1 in peripheral blood and after in vitro stimulation with anti-TNF antibody. MATERIAL AND METHODS: 55 patients with diabetes type 1 were examined. Mean duration of the disease in the diabetic group was 6.45 (+/-3.7) years. The percentage of Tregs and Tregs carrying CD62L was evaluated using flow cytometry in peripheral blood of diabetic patients as well as after in vitro stimulation with the anti-CD3 (control), the anti-CD3 with the anti-TNF and anti-CD3 with TNF. RESULTS: Diabetic type 1 patients were characterized by a lower percentage of Tregs and Tregs expressing CD62L subset compared to their healthy counterparts. In addition, these cells reacted differently to anti-CD3 antibody stimulation than the cells from the healthy individuals. Anti-TNF induced a significant increase in the percentage of CD4+Foxp3+ and CD4+Foxp3+CD62Lhigh regulatory T cells. After treatment with TNF-alpha the frequency of these cells significantly decreased. In the diabetic group we showed a negative correlation between CD4+CD25highCD62Lhigh T lymphocytes and HbA1c [r=(-0.25)] as well as CRP level [r=(-0.38)]. CONCLUSIONS: The lower percentage of Tregs and CD62Lhigh Tregs may contribute to ineffective suppression of proinflammatory cytokine production during type 1 diabetes. We suggest the protective role of anti-TNF on Treg subset in diabetic type 1 patients and we highlight the importance of proinflammatory cytokine - TNF-alpha, which may be responsible for quantitative abnormalities in Treg subset in these patients.

Immunosenescence increases the rate of acceptance of kidney allotransplants in elderly recipients through exhaustion of CD4+ T-cells.

Compromised immunity is the hallmark of ageing. Paradoxically, it may be "an ally" in facilitating acceptance of allogeneic grafts in the elderly. In this retrospective study we looked for biomarkers of immunosenescence that distinguish elderly recipients less prone to reject kidney allografts. Recruited kidney recipients aged > or = 60 or < 60 were designated 'elderly' and 'young', respectively. Both age-groups were divided according to the history of acute rejection. The phenotype, length of telomeres, expression of FoxP3 and proliferative responses were assessed in CD4(+) and CD8(+) T-cell subsets. In addition, IL6, IL10 and TGFbeta were measured on the level of mRNA and serum protein. Acute-rejection-free history in elderly transplant recipients was associated with short telomeres, a decreased proportion of CD28(+) T-cells associated with CMV-seropositivity and low proliferation of CD4(+) T-cells. In contrast, elderly recipients who experienced acute rejection kept preserved telomere length, had a higher number of functional CD4(+)CD28(+) cells and exhibited vigorous proliferation in vitro. These differences were not found in the young group. The major conclusion of this study is that the impaired condition of CD4(+) T-cells, so-called immunosenescence, renders transplant recipients less responsive to an allogeneic kidney graft, an effect that was limited to transplant recipients of > 60 years of age.

Interleukin-2 as a predictor of early postoperative atrial fibrillation after cardiopulmonary bypass graft (CABG).

Recently, inflammation has been considered as a risk factor of postoperative atrial fibrillation (PAF). The main purpose of this study was to estimate the connections between occurrence of PAF and cytokine release. Thirty-three patients who qualified for cardiopulmonary bypass graft (CABG) were included in the study. Blood was taken from all of them before CABG, then 3 h, 24 h, and 72 h afterwards. Cytokine (IL-6, IL-2, IL-4, IL-10, IFN-gamma, TNF-alpha) concentration was measured at every time point. Eleven patients developed atrial fibrillation after the CABG. Five of them developed PAF until 1 day post-CABG and six of them after 1 day post-CABG. Patients who developed PAF before 1 day post-CABG were characterized by a higher level of IL-2 in sera before 24 h and 72 h post-CABG compared with patients without PAF. Moreover, the PAF before 1 day post-CABG group was also characterized by the higher level of IFN-gamma and IL-10 at 24 h after intervention. Analysis of patients who developed PAF after 1 day post-CABG revealed a higher level of IL-10 and IFN-gamma at 24 h post-CABG compared with patients without PAF. In this study, we have shown for the first time a straightforward connection between IL-2 sera levels and the development of PAF shortly after CABG.

Inflammatory response of coronary artery disease postmenopausal women is associated with the IVS1-397T > C estrogen receptor alpha polymorphism.

We analyzed the relationship between the IVS1-397T > C polymorphism of estrogen receptor alpha gene and inflammatory response of eighty postmenopausal women with an established coronary artery disease (CAD). We found that the IVS1-397T allele carriers exhibited an enhanced inflammatory response in vivo and in vitro. These patients had a higher number of total monocytes and their CD14(+)CD16(+) inflammatory subset in relation to CC homozygotes. The CT and TT women's LPS-stimulated whole blood cell cultures produced more IL6 and TNFalpha than did the cultures of CC women. Moreover, significantly more of the T allele patients had major post-coronary artery bypass grafting complications and less of them experienced subjective angina pectoris improvement. Summarizing, the IVS1-397T > C polymorphism allows us to identify a group of postmenopausal women with the strong inflammatory response which is associated with a higher incidence of the major post-CABG adverse cardiovascular complications.

Serum TNF-alpha level predicts nonproliferative diabetic retinopathy in children.

The aim of this study was identification of the immunologic markers of the damage to the eye apparatus at early stages of diabetes mellitus (DM) type 1 children. One hundred and eleven children with DM type 1 were divided into two groups: those with nonproliferative diabetic retinopathy (NPDR) and without retinopathy. All the children had their daily urine albumin excretion, HbA1c, C-peptide measured, 24-hour blood pressure monitoring, and ophthalmologic examination. Levels of TNF-alpha, IL-6, and IL-12 in serum were measured by ELISA tests (Quantikine High Sensitivity Human by R&D Systems, Minneapolis, Minn, USA). The NPDR children demonstrated a significantly longer duration of the disease in addition to higher HbA1c, albumin excretion rate, C-reactive protein, systolic blood pressure, as well as TNF-alpha and IL-6 levels than those without retinopathy. The logistic regression revealed that the risk of NPDR was strongly dependent on TNF-alpha [(OR 4.01; 95%CI 2.01-7.96)]. TNF-alpha appears to be the most significant predictor among the analyzed parameters of damage to the eye apparatus. The early introduction of the TNF-alpha antagonists to the treatment of young patients with DM type 1 who show high serum activity of the TNF-alpha may prevent them from development of diabetic retinopathy.

NK cell compartment in patients with coronary heart disease.

BACKGROUND: Viral and bacterial infections have been considered as a risk factor for Coronary Heart Disease (CHD). NK cells, as a first line of defense against those infections, may play a role in CHD development. Thus, the main aim of our study was to determine NK cell compartment in patients with CHD undergoing coronary artery by-pass grafting. RESULTS: Ninety three patients with CHD were included into the study; the control group consisted of 49 healthy volunteers. As compared to controls, CHD patients had lower NK cytotoxic activity. CHD group had also a decreased absolute number and percentage of total NK cells and CD3-CD56dim cytotoxic NK subset. In addition, we observed tendency toward lower percentage of the CD3-CD56bright regulatory NK subset and CD3-CD56+IFN-gamma+ cells in CHD patients. CONCLUSION: These data indicate that CHD is associated with an impairment of NK cells compartment.

Long-term prognosis after coronary bypass surgery depends on interleukin 6 polymorphism and past acute infections.

We were seeking for a mutual link between the -174G>C IL6 promoter polymorphism, history of the past acute respiratory infections and the long-term post-coronary artery bypass grafting (CABG) major adverse cardiovascular events (MACE) incidence. Two hundred thirty seven post-CABG patients have been followed up for a median period of 36 months. We found that past acute infections, influenza-like illness and lack of vaccination against influenza confer a significant risk of the post-CABG MACE incidence in the -174G allele carrying patients.

Interleukin 6 polymorphism corresponds to the number of severely stenosed coronary arteries.

IL6 gene promoter polymorphisms may influence the outcome of cardiovascular diseases. The aim of our study was to find out whether the -174G>C polymorphism, as well as the IL6 secretory profile, may be linked to the number of severely (> or = 75%) occluded coronary arteries in patients with advanced coronary heart disease (CHD). Three hundred and twenty patients awaiting elective coronary artery bypass grafting were enrolled into the study. Blood was taken the day before surgery. The PCR-RFLP method was used for IL6 gene polymorphism analysis. Spontaneous IL6 release was measured by bioassay in supernatants of whole blood cell cultures (WBCC) incubated for 24 h and 48 h. We found that significantly more patients with triple vessel disease were found within the -174GG group as compared to the -174GC and CC genotype carriers. The highest IL6 serum levels were found in the -174GG and the lowest in the -174CC genotype patients. Spontaneous in vitro IL6 secretion appeared to be significantly higher at all time points in the -174GG as compared to the CC and GC genotype carriers. The serum concentration of IL6 and the spontaneous IL6 secretion were directly related to the number of obstructed coronary vessels. Our results emphasize the role of IL6 as an important, non-classical risk factor predicting the number of severely affected coronary vessels.

Effects of intranasal 17beta-estradiol administration on serum bioactive interleukin-6 and C-reactive protein levels in healthy postmenopausal women.

OBJECTIVE: Oral estrogen increases the levels of C-reactive protein (CRP), which is an independent risk factor for coronary heart disease in healthy individuals. The aim of our study was to investigate the effects of intranasal 17beta-estradiol (E2) on serum CRP and its most potent stimulant, interleukin-6 in healthy postmenopausal women. DESIGN: Thirty-six healthy postmenopausal women (45-54 y) were enrolled. According to their individual preferences, they were assigned to intranasal (n = 10), transdermal (n = 14), or oral (n = 12) continuous E2 treatment with a sequential progestin (10-14 d in a 28-d cycle). Blood samples were drawn at baseline and after 3, 6, and 12 months during the estrogen-only phase to adjust for the progestin effect. RESULTS: In women taking intranasal or transdermal E2, there were no significant changes in median serum CRP levels during the 12-month treatment period. In women taking oral E2 preparations, serum median CRP levels were significantly higher compared to baseline after 6 and 12 months of the therapy (P < 0.05). Conversely, serum median bioactive interleukin-6 levels were significantly lower after 6 and 12 months in women taking E2 intranasally or orally and after 12 months in women taking E2 transdermally (P < 0.05). CONCLUSIONS: The results of our study show that intranasal, similarly to transdermal, E2 administration does not increase serum CRP levels in postmenopausal women. They also support the hypothesis that CRP increase during oral estrogen treatment is not mediated by the enhancement of interleukin-6 production by the immune cells but is rather caused by the hepatic first-pass metabolism effect.

High serum interleukin-18 concentrations in patients with coronary artery disease and type 2 diabetes mellitus.

AIMS: The aim of our study was to analyse the serum level of interleukin 18 (IL-18) in coronary artery disease (CAD) patients with type 2 diabetes mellitus (DM), and to relate this to clinical findings. METHODS: The IL-18 level was measured by ELISA in serum samples from 130 CAD patients prior to their first, elective, coronary artery bypass surgery. Forty-three of them had been diabetic for several years. A control group consisted of 31 healthy people matched according to age, BMI, lipid and smoking status. RESULTS: The CAD patients with DM were similar to the non-diabetic CAD patients with respect to age, BMI, grade of heart failure, ejection fraction. There were no differences in the duration of CAD, history of myocardial infarction and PTCA or instability of angina. The serum level of IL-18 was higher in the CAD patients than in the control group. The CAD patients with DM had a higher concentration of IL-18 compared to the non-diabetic CAD group. The diabetic patients with triple-vessel disease were characterized by a higher concentration of IL-18 than the non-diabetic patients with the same grade of CAD. Smoking affected the IL-18 concentration, particularly in the diabetic patients. CONCLUSION: Type 2 DM predisposes patients, especially those with multi-vessel CAD who were smokers, to a higher serum level of IL-18, which may help explain their vulnerability to fatal, secondary cardiovascular events. These patients should be in the first line for stringent, secondary cardiovascular prevention.

Long-term therapy with recombinant human erythropoietin increases CD8+ T-cell apoptosis in haemodialysis patients.

BACKGROUND: We intended to assess the intensity of apoptosis in the CD4+ and CD8+ T-lymphocytes of haemodialysis (HD) patients on recombinant human erythropoietin (rHuEpo). METHODS: The expression of Fas, tumour necrosis factor-alpha receptors (TNFRI and TNFRII) and the CD28 molecule on lymphocytes was evaluated in 15 HD patients before and during treatment with rHuEpo. In cultures of peripheral blood mononuclear cells (PBMCs) stimulated with rHuEpo, phytohaemagglutinin and camptothecin, our measures of apoptosis were the percentages of cells with subdiploid DNA content and of annexin V-stained cells. Results, Therapy with rHuEpo did not affect CD4+ T cells but decreased the percentage of CD8+ T cells in peripheral blood. The intensity of apoptosis in both CD4+ and CD8+ T cells at baseline was lower in HD patients than in healthy volunteers, and increased in those treated with rHuEpo. In vitro, rHuEpo did not induce apoptosis in PBMCs. The percentage of CD8+Fas+ T cells was constant, while that of CD8+TNFRI+ cells declined during follow-up. There was an increase in the percentage of CD28+ T cells, mainly in the CD8+ compartment, as early as 1 month after the introduction of rHuEpo. CONCLUSIONS: Treatment with rHupo caused a decline of CD8+ T cells in HD patients, which most probably was mediated via the TNFRI-related apoptotic pathway and was independent of Fas expression. Apoptosis in vitro was not directly influenced by rHuEpo, suggesting that the process in vivo was only initiated by rHuEpo supplementation.

No effects of ozonated autohemotherapy on inflammation response in hemodialyzed patients.

BACKGROUND: Ozone as a strong oxidant may induce an inflammatory response. AIM: The hypothesis was verified as to whether ozonated autohemotherapy using an ozone dose in therapeutic range changes the plasma concentration of C-reactive protein and interleukin-6, markers of inflammation. METHODS: In a controlled, single-blind, cross-over study, 12 chronically hemodialyzed patients with peripheral arterial disease were exposed to nine sessions of autohemotherapy with blood exposure to oxygen as a control followed by nine sessions of ozonated autohemotherapy with an ozone concentration of 50 microg/ml. RESULTS: There was no statistical difference between C-reactive protein levels at baseline (1.53 +/- 1.01 mg/l), after nine sessions of control autohemotherapy (1.48 +/- 0.96 mg/l), and after nine sessions of ozonated autohemotherapy (1.55 +/- 0.84 mg/l). There was also no statistical difference between the interleukin-6 serum concentration at baseline (438 +/- 118 pg/ml), after nine sessions of control autohemotherapy (444 +/- 120 pg/ml), and after nine sessions of ozonated autohemotherapy (466 +/- 152 pg/ml). CONCLUSION: The results of this study suggest that ozonated autohemotherapy using an ozone concentration of 50 microg/ml does not induce an inflammatory response.