Podiatric care from diabetologists point of view.

Scoring for the risk of Diabetic foot syndrome (DFS) should be performed regularly in each patient with diabetes mellitus (DM). Patients at risk for DFS should be followed by diabetologists, those with moderate and severe risk for the development of DFS or those with DFS in remission should be already followed by podiatrists. The aim of our study was to determine the extent of DFS risk screening procedures, dispensary care of patients at risk for DFS and treatment of patients with newly developed DFS in diabetes clinics in the Czech Republic. METHODS: To find out the study data, we prepared in cooperation with the ČDS ČLS JEP Committee a questionnaire survey for outpatient diabetology specialists. RESULTS: The questionnaire was completed by 57% (76/135) of diabetologists. Most of them dispensary approximately 1000- 2000 patients with DM. Their feet are checked by 98.7% of diabetologists (1.6 ± 0.8 times a year on average). Screening for the risk of DFS (13024) is performing in less than 100 patients by 74.3% of diabetologists, in 100-200 patients by 14.9% and in more than 200 patients by 10.8% of diabetologists. 77% of respondents are able to examine neuropathy, the rest send their patients to neurologists, peripheral arterial disease is evaluated by only 47.3% of diabetologists (35.3% of them use some form of instrumental examination), others (48.6%) send patients to angiologists, 4.1% of diabetologists do not examine PAD at all). Based on the assessed findings, more than half of the respondents (50.7%) perform scoring for the risk of DFS, but 1/5 of outpatient diabetologists do not know how the scoring is performed. If colleagues find a patient at a risk for DFS, they usually follow him/her by themselves (64.4%), in 24.6% of cases they send the patient immediately to podiatry or surgery (11%). If a patient with a new DFS comes at diabetology clinic, 72.6% of diabetologists are able to prescribe off-loading, 60.3% antibiotics, 47.9% local therapy. Only 52.1% of diabetologists send a patient with a new DFS to outpatient foot clinic, 39.7% to surgery, the rest of them elsewhere. CONCLUSION: Based on the questionnaire survey results, the screening of DFS is currently severely undersized in outpatient diabetology clinics, it is sufficiently performed only by 11% of diabetologists. Only 16% of diabetologists perform some form of non-invasive diagnostic procedures detecting peripheral arterial disease, neuropathy examinations are more common. If a diabetologist meet a patient with newly developed DFS, he/she is able to prescribe off-loading or antibiotics, but only half of the diabetologists send the patient to outpatient foot clinic, probably due to a lack of them or their overload.

Stimulation TcPO2 Testing Improves Diagnosis of Peripheral Arterial Disease in Patients With Diabetic Foot.

Background: All diagnostic procedures of peripheral arterial disease (PAD) in diabetic foot (DF) are complicated due to diabetes mellitus and its late complications.The aim of our study is to enhance diagnosis of PAD using a novel transcutaneous oximetry (TcPO2) stimulation test. Methods: The study comprised patients with mild-to-moderate PAD(WIfI-I 1 or 2) and baseline TcPO2 values of 30-50 mmHg.TcPO2 was measured across 107 different angiosomes. Stimulation examination involved a modification of the Ratschow test. All patients underwent PAD assessment (systolic blood pressures (SBP), toe pressures (TP), the ankle-brachial indexes (ABI) and toe-brachial indexes (TBI), duplex ultrasound of circulation). Angiosomes were divided into two groups based on ultrasound findings: group M(n=60) with monophasic flow; group T(n=47) with triphasic flow. Large vessel parameters and TcPO2 at rest and after exercise (minimal TcPO2, changes in TcPO2 from baseline (Δ,%), TcPO2 recovery time) measured during the stimulation test were compared between study groups. Results: During the TcPO2 stimulation exercise test, group M exhibited significantly lower minimal TcPO2 (26.2 ± 11.1 vs. 31.4 ± 9.4 mmHg; p<0.01), greater Δ and percentage decreases from resting TcPO2 (p=0.014 and p=0.007, respectively) and longer TcPO2 recovery times (446 ± 134 vs. 370 ± 81ms;p=0.0005) compared to group T. SBPs, TPs and indexes were significantly lower in group M compared to group T. Sensitivity and specificity of TcPO2 stimulation parameters during PAD detection increased significantly to the level of SBP, ABI, TP and TBI. Conclusion: Compared to resting TcPO2, TcPO2 measured during stimulation improves detection of latent forms of PAD and restenosis/obliterations of previously treated arteries in diabetic foot patients. Clinical Trial Registration: ClinicalTrials.gov [https://register.clinicaltrials.gov/prs/app/action/SelectProtocol?sid=S0009V7W&selectaction=Edit&uid=U0005381&ts=2&cx=3j24u2], identifier NCT04404699.

Insulin Management of Patients with Inadequately Controlled Type 2 Diabetes Admitted to Hospital: Titration Patterns and Frequency of Hypoglycemia as Results of a Prospective Observational Study (Hospital Study).

INTRODUCTION: Despite the continuously growing number of therapeutic options for type 2 diabetes mellitus (T2DM) including insulins, a large percentage of patients fail to achieve HbA1c targets. Several real-world studies focused on patients with T2DM receiving insulin treatment in outpatient settings were conducted, but information about real-world in-hospital insulin management is lacking. The aim of this study was to describe the management of insulin therapy with a focus on basal-bolus and premixed insulin regimens in patients with T2DM under routine in-hospital medical practice in the Czech Republic. METHODS: This non-interventional prospective study was conducted from June 2014 to December 2017 in 22 centers in the Czech Republic under routine clinical practice conditions. Adult patients admitted to hospital with metabolically uncontrolled T2DM [HbA1c ≥ 60 mmol/mol; > 7.6% Diabetes Control and Complications Trial (DCCT)] and there treated with basal-bolus and premixed insulin regimens were documented during hospitalization. RESULTS: Overall, 369 patients with T2DM (54.7% male, mean age 64.44 ± 13.84 years, BMI 31.10 ± 6.00 kg/m2, duration of diabetes 8.11 ± 9.93 years, HbA1c 95.90 ± 24.38 mmol/mol, length of stay was 7.94 ± 4.53 days) were included. The percentage of glucose values under 10 mmol/l at time of randomization (the group with basal-bolus insulin regimen vs. the premix insulin regimen group) was 24.2% vs. 33.5% (p = 0.053), at time of first insulin dose adjustment it was 43.1% vs. 50.0% (p = 0.330), and 1 day before hospital discharge it was 61.7% vs. 61.4% (p = 0.107). A hypoglycemic event occurred in a total of 15 patients in the basal-bolus regimen group, and no hypoglycemic event occurred in the premixed insulin regimen group. CONCLUSION: In-hospital insulin management regarding basal-bolus and premixed insulin regimens is safe and in concordance with current international recommendations.

Direct Versus Iterated Multi-Step Forecasting of Glycaemia in Type 1 Diabetics Using Autoregressive Models.

The paper compares two approaches to multi-step ahead glycaemia forecasting. While the direct approach uses a different model for each number of steps ahead, the iterative approach applies one one-step ahead model iteratively. Although it is well known that the iterative approach suffers from the error accumulation problem, there are no clear outcomes supporting a proper choice between those two methods. This paper provides such comparison for different ARX models and shows that the iterative approach outperformed the direct method for one-hour ahead (12-steps ahead) forecasting. Moreover, the classical linear ARX model outperformed more complex non-linear versions for training data covering one-month period.

Ensemble methods in combination with compartment models for blood glucose level prediction in type 1 diabetes mellitus.

Backgroung: Type 1 diabetes is a disease that adversely affects the daily life of a large percentage of people worldwide. Daily glucose levels regulation and useful advices provided to patients regarding their diet are essential for diabetes treatment. For this reason, the interest of the academic community has focused on developing innovative systems, such as decision support systems, based on glucose prediction algorithms. The present work presents the predictive capabilities of ensemble methods compared to individual algorithms while combining each method with compartment models for fast acting insulin absorption simulation. Methods: An approach of combining widely used glycemia prediction algorithms is proposed and three different ensemble methods (Linear, Bagging and Boosting metaregressor) are applied and evaluated on their ability to provide accurate predictions for 30, 45 and 60 minutes ahead prediction horizon. Moreover, glycemia levels, long and short acting insulin dosages and consumed carbohydrates from six type one people with diabetes are used as input data and the results are evaluated in terms of root-mean square error and Clarke error grid analysis. Results: According to results, ensemble methods can provide more accurate glucose concentration in comparison to individual algorithms. Bagging metaregressor, specifically, performed better than individual algorithms in all prediction horizons for small datasets. Bagging ensemble method improved the percentage in zone A according to Clarkes error grid analysis by 4% and in some cases by 9%. Moreover, compartment models are proved to improve results in combination with any method at any prediction horizon. This strengthen the potential practical usefulness of the ensemble methods and the importance of building accurate compartment models.

Ethanol and risk of hypoglycemia in patients with type 1 diabetes mellitus: a brief overview for clinical practice.

Alcoholic drinks are one of the risk factors for hypoglycemia. Ethanol inhibits gluconeogenesis, decreases a level of growth hormone and impairs hypoglycemia awareness. The risk of hypoglycemia while drinking alcohol can be reduced by parallel ingestion of food (saccharides). Some recommendations also mention the change of insulin doses.

Comprehensive Analysis of the Real Lifestyles of T1D Patients for the Purpose of Designing a Personalized Counselor for Prandial Insulin Dosing.

Post-prandial hyperglycemia is still a challenging issue in intensified insulin therapy. Data of 35 T1D patients during a four-week period were analyzed: RT-CGM (real time continuous glucose monitoring) record, insulin doses, diet (including meal photos), energy expenditure, and other relevant conditions. Patients made significant errors in carbohydrate counting (in 56% of cooked and 44% of noncooked meals), which resulted in inadequate insulin doses. Subsequently, a mobile application was programmed to provide individualized advice on prandial insulin dose. When using the application, a patient chooses only the type of categorized situation (e.g., meals with other relevant data) without carbohydrates counting. The application significantly improved postprandial glycemia as normoglycemia was reached in 95/105 testing sessions. Other important findings of the study include: A high intake of saturated fat (median: 162% of recommended intake); a low intake of fiber and vitamin C (median: 42% and 37%, respectively, of recommended intake); an increase in overweight/obesity status (according to body fat measurement), especially in women (median of body fat: 30%); and low physical activity (in 16/35 patients). The proposed individualized approach without carbohydrate counting may help reach postprandial normoglycemia but it is necessary to pay attention to the lifestyle habits of T1D patients too.

Analysis of Postprandial Glycemia in Relation to Metabolic Compensation and Other Observed Parameters of Outpatients with Type 2 Diabetes Mellitus in the Czech Republic.

INTRODUCTION: The goal of the study was to determine the level of metabolic compensation expressed by glycosylated hemoglobin, fasting plasma glucose, and postprandial glucose as determined after a standardized breakfast; further, to evaluate interrelationships between the studied parameters and postprandial glucose levels. METHODS: The study included 1055 patients with type 2 diabetes mellitus. Their fasting plasma glucose and postprandial glucose were measured before and after a standardized breakfast. Attending diabetologists completed a uniform questionnaire that included demographic data, type of antidiabetic treatment, duration of diabetes, latest glycosylated hemoglobin value, presence of dyslipidemia, and organic complications. RESULTS: Glycosylated hemoglobin < 53 mmol/mol was achieved in 363 (34.2%), postprandial glucose < 7.5 mmol/l in 211 (19.9%), and fasting plasma glucose < 6 mmol/l in 251 (23.7%) patients. Excellent metabolic compensation, indicated by all the above mentioned glycosylated hemoglobin, fasting plasma glucose, and postprandial glucose values simultaneously, was achieved in only 71 (6.7%) patients. Comparable to fasting plasma glucose and postprandial glucose values, correlation with glycosylated hemoglobin levels is statistically significant; however, there is no difference at different glycosylated hemoglobin levels. There was a significant correlation between dyslipidemia and postprandial glycemia (p = 0.013). CONCLUSION: The objective of care for patients with diabetes mellitus is to improve their long-term metabolic compensation; to that end, both fasting plasma glucose and postprandial glucose deserve equal attention.

Cardiac autonomic neuropathy may play a role in pathogenesis of atherosclerosis in type 1 diabetes mellitus.

AIMS: Cardiac autonomic neuropathy (CAN) is a frequent and severe complication of type 1 diabetes mellitus (T1DM). CAN diagnosis is associated with increased cardiovascular morbidity and mortality, often due to progressive atherosclerosis. Carotid intima media thickness (CIMT) is a surrogate marker of the atherosclerosis. The aim of our study was to evaluate the relationship between CIMT and CAN in T1DM patients. METHODS: Total of 49 T1DM patients and 45 healthy controls were examined for CAN presence and CIMT. CAN was diagnosed based on the results of Ewing test battery and spectral analysis of heart rate variability. CIMT was measured by two-dimensional ultrasound. Biochemical, anthropometric and anamnestic risk markers of atherosclerosis were evaluated. We used logistic types of generalized additive models (GAM) for statistical analysis. RESULTS: CAN was detected in 22 out of 49 T1DM patients (45%). All 45 healthy controls had normal cardiovascular autonomic tests results. CIMT was significantly positively associated with T1DM diagnosis (p=0.0251), CAN diagnosis (p=0.007), age (p<0.0001), BMI (p=0.0435) and systolic blood pressure (p=0.0098). CAN effect on CIMT interacted with the effect of T1DM. The combination of both factors significantly increased CIMT more than the sum of the individual T1DM and CAN status. CONCLUSIONS: CAN is significantly associated with higher CIMT in T1DM patients. CAN may play a role in pathogenesis of atherosclerosis in type 1 diabetes mellitus.

Not Only Glycaemic But Also Other Metabolic Factors Affect T Regulatory Cell Counts and Proinflammatory Cytokine Levels in Women with Type 1 Diabetes.

Type 1 diabetic (T1D) patients suffer from insulinopenia and hyperglycaemia. Studies have shown that if a patient's hyperglycaemic environment is not compensated, it leads to complex immune dysfunctions. Similarly, T1D mothers with poor glycaemic control exert a negative impact on the immune responses of their newborns. However, questions concerning the impact of other metabolic disturbances on the immune system of T1D mothers (and their newborns) have been raised. To address these questions, we examined 28 T1D women in reproductive age for the relationship between various metabolic, clinical, and immune parameters. Our study revealed several unexpected correlations which are indicative of a much more complex relationship between glucose and lipid factors (namely, glycosylated haemoglobin Hb1Ac, the presence of one but not multiple chronic diabetic complications, and atherogenic indexes) and proinflammatory cytokines (IL-1alpha and TNF-alpha). Regulatory T cell counts correlated with HbA1c, diabetic neuropathy, lipid spectra parameters, and IL-6 levels. Total T-helper cell count was interconnected with BMI and glycaemia variability correlated with lipid spectra parameters, insulin dose, and vitamin D levels. These and other correlations revealed in this study provide broader insight into the association of various metabolic abnormalities with immune parameters that may impact T1D mothers or their developing child.

[Impaired hypoglycemia awareness in diabetes mellitus]. / Syndrom poruseného vnímání hypoglykemie u diabetes mellitus.

UNLABELLED: Impaired hypoglycemia awareness is defined at the onset of neuroglycopenia without the appearance of autonomic warning symptoms. Impaired hypoglycemia awareness is disorder which affects aprox. one third of patients with type 1 diabetes mellitus and 8-10 % of patients with type 2 diabetes mellitus who are treated with insulin. The most dangerous consequence is 6 times higher frequency of severe hypoglycemia in patients with Type 1 diabetes mellitus and 17 times higher in Type 2 diabetic patients treated with insulin. Treatment of impaired hypoglycemia awareness is complex, based on a multifactorial intervention of clinical care and structured patient education. KEY WORDS: diabetes mellitus - hypoglycemia- impaired hypoglycemia awareness.

[Incidence of amputations among patients with diabetes mellitus in the Czech Republic from 2010 to 2014]. / Incidence amputací u pacientu s diabetes mellitus v Ceské republice v letech 2010-2014.

INTRODUCTION: Information about the incidence of organ-affecting complications of diabetes, including the diabetic foot syndrome, can be obtained from the documents of the Institute of Health Information in the Czech Republic. GOAL: Assessment of the development of high amputations and minor surgical procedures on the lower limb from 2010 to 2014 in a representative sample of the population of patients with DM kept in the General Health Insurance Company of the Czech Republic database. METHODOLOGY: We identified all individuals in the VZP database who had a record of DM diagnosis (E10-E16 based on ICD 10) or any antidiabetic therapy prescribed (ATC group A10) in the period of 2010-2014. A set of patients who had an agent from A10 group prescribed at least once in the given year was extracted for analysis. In the next step we identified individuals, who in the period of 2010-2014 also underwent a surgical procedure on the lower limb due to diabetic foot. RESULTS: An absolute number of lower limb amputations remains at a stationary level. CONCLUSION: The submitted analysis presents the first assessment of the development of surgical treatment of diabetic foot in the Czech Republic. The amount of surgical procedures on the diabetic foot remains stable, regarding both high amputations and lower limb minor surgical procedures. In the context of an absolute increase of patients treated for diabetes mellitus, the stationary state is an indication of a relative decrease, which is favourable in particular with regard to the amputation of long bones.

[The PROROK project results after 6 months of intervention (Prospective observation project focusing on the relevance of the difference between fasting blood glucose levels and postprandial blood glucose for estimation of success of type 2 diabetes therapy)]. / Výsledky projektu PROROK po 6 mesících intervence (Prospektivní observacní projekt významu diference glykemie nalacno a postprandiální glykemie pro odhad úspesnosti terapie diabetu 2. typu).

INTRODUCTION: The PROROK project (Prospective observation project focusing on the relevance of the difference between fasting and postprandial blood glucose levels for the estimation of success of type 2 diabetes therapy) had a character of non-interventional, prospective, multicentric observation study lasting 6 months, whose goal was to quantify the relevance of the difference between fasting and postprandial blood glucose levels to the success of the treatment with GLP1 receptor agonists, resp. the treatment with basal, premixed insulin, or a combination of basal-bolus insulin. Physicians chose a therapy for patients with insufficiently compensated problems as they considered appropriate; 4,972 patients were included. GOAL: Evaluation of the intervention results for the patients included in the PROROK observation project with a focus on the choice of therapy by the treating diabetologist after 6 months of observation. RESULTS: An average improvement of the glycated hemoglobin values in the whole cohort reached 1.6%, the median of the resulting glycated hemoglobin reached 5.9% and 5.8% resp. (basal insulin). Statistically significant was the change in the median weight in the cohort treated with GLP-1 receptor agonists, from 105 kg to 100 kg; this did not significantly change in the other cohorts. The change of waist circumference over time in all patients and in the individual cohorts was consistent with the change of weight. The median change of fasting blood glucose levels in the whole cohort was -1.7 mmol/l after 3 months and -2.4 mmol/l (p<0.001) after 6 months. The greatest absolute decrease was recorded in the cohort treated with basal insulin (-2.8 mmol/l). The median change of postprandial blood glucose levels was -2.4 mmol/l after 3 months and -3.3 mmol/l (p<0.001) after 6 months. The greatest absolute decrease was recorded in the branch treated with a combination of prandial and basal insulin (-3.9 mmol/l). All differences p<0.001. CONCLUSION: The choice of therapy in the PROROK project is in agreement with the basic findings in pathophysiology of type 2 diabetes and with the options of an individually chosen targeted intervention involving antidiabetic therapy. The results of the six-month observation have proven the individual choice of therapy correct. In the cohort of diabetic patients differing at the beginning in weight, waist circumference, fasting blood glucose and the difference between fasting and postprandial glucose levels, an individually chosen therapy led to the same final result, while an absolute change in the followed parameters differed in the individual groups.

[Do diabetologists choose a therapy rationally? Basic results of the PROROK project (a prospective observation project to assess the relevance of the difference between fasting glycemia and postprandial glycemia to estimation of success of type 2 diabetes therapy)]. / Volí diabetologové terapii racionálne? Základní výsledky projektu PROROK (Prospektivní observacní projekt významu diference glykemie nalacno a postprandiální glykemie pro odhad úspesnosti terapie diabetu 2. typu).

INTRODUCTION: The PROROK project (A prospective observation project to assess the relevance of the difference between fasting glycemia and postprandial glycemia to estimation of success of type 2 diabetes therapy) had a character of a non-interventional, prospective, multicentric observation project conducted for a period of 6 months, whose aim was to quantify the relevance of the difference between fasting and postprandial glycemia to the success of GLP1 receptor agonist treatment, or insulin therapy with basal or premixed insulin, or a combination of basal and bolus insulin. Physicians chose therapy for inadequately compensated patients at their own discretion, with 4 972 patients included. AIM: The study aimed at the assessment of the differences in basic anthropometric and biochemical parameters between the patient cohorts included in the PROROK project with regard to the therapy selected by the treating diabetologist. METHODOLOGY AND RESULTS: The patients treated with GLP1 receptor agonists were quite young, they have suffered from diabetes for a shorter period of time and at the same time were more obese and had the highest concentration of triacylglycerols. The patients who underwent basal insulin therapy, had the highest fasting glycemia. The patients for whom premixed insulin therapy or basal/bolus insulin regimen were chosen, manifested the highest postprandial glycemia, those with basal/bolus insulin regimen had the highest initial glycated haemoglobin. The difference between fasting and postprandial glycemia was the smallest in the cohort for which basal insulin therapy was chosen and the greatest in the cohort chosen for the therapy with premixed insulin, or with the basal/bolus insulin combination. Average improvement in glycated haemoglobin values reached 1.6 % within the whole cohort, a median of the resulting glycated haemoglobin reached 5.9 % or 5.8 % (GLP1 receptor agonist treatment). All the differences amounted to p < 0.001. CONCLUSION: Bearing in mind that the differences established in the parameters describing the cohorts, although statistically relevant, are of smaller clinical relevance, we regard as an important finding that the choice of therapy is in accordance with the basic knowledge about the pathophysiology of type 2 diabetes and possibilities of an individually chosen targeted intervention with antidiabetic therapy. We may conclude that most of the physicians participating in the PROROK project choose their therapy in a rational manner.

The effect of diabetes-associated autoantigens on cell processes in human PBMCs and their relevance to autoimmune diabetes development.

Type 1 Diabetes (T1D) is considered to be a T-helper- (Th-) 1 autoimmune disease; however, T1D pathogenesis likely involves many factors, and sufficient tools for autoreactive T cell detection for the study of this disease are currently lacking. In this study, using gene expression microarrays, we analysed the effect of diabetes-associated autoantigens on peripheral blood mononuclear cells (PBMCs) with the purpose of identifying (pre)diabetes-associated cell processes. Twelve patients with recent onset T1D, 18 first-degree relatives of the TD1 patients (DRL; 9/18 autoantibody positive), and 13 healthy controls (DV) were tested. PBMCs from these individuals were stimulated with a cocktail of diabetes-associated autoantigens (proinsulin, IA-2, and GAD65-derived peptides). After 72 hours, gene expression was evaluated by high-density gene microarray. The greatest number of functional differences was observed between relatives and controls (69 pathways), from which 15% of the pathways belonged to "immune response-related" processes. In the T1D versus controls comparison, more pathways (24%) were classified as "immune response-related." Important pathways that were identified using data from the T1D versus controls comparison were pathways involving antigen presentation by MHCII, the activation of Th17 and Th22 responses, and cytoskeleton rearrangement-related processes. Genes involved in Th17 and TGF-beta cascades may represent novel, promising (pre)diabetes biomarkers.

Protein microarray analysis as a tool for monitoring cellular autoreactivity in type 1 diabetes patients and their relatives.

BACKGROUND: Autoreactive T cells have a crucial role in type 1 diabetes (T1D) pathogenesis. OBJECTIVES: The aim of our study was to monitor the in vitro production of cytokines by peripheral blood mononuclear cells (PBMCs) after stimulation with diabetogenic autoantigens. SUBJECTS: Ten T1D patients (tested at the time of diagnosis and 6 and 12 months later), 10 first-degree relatives of the T1D patients, and 10 controls underwent the study. METHODS: PBMCs were stimulated with glutamic acid decarboxylase 65 (GAD65) amino acids (a.a.) 247-279, 509-528, and 524-543; proinsulin a.a. 9-23; and tyrosine phosphatase (islet antigen-2)/R2 a.a. 853-872. Interleukin (IL)-2, IL-4, IL-5, IL-6, IL-10, IL-13, interferon (IFN)-gamma, tumor necrosis factor beta, transforming growth factor beta1, and granulocyte colony-stimulating factor (GCSF) were analyzed by protein microarray. RESULTS: Differences in cytokine(s) poststimulatory and mainly in basal production were observed in all groups. The most prominent findings were in controls, the higher basal levels of IL-2, IL-4, IL-5, IL-13, and GCSF were observed when compared with relatives (p < 0.05, for all). After stimulation in controls, there was a significant decrease in IL-2, IL-13, GCSF, and IFN-gamma (p < 0.05, for all). The group of relatives was the most variable in poststimulatory production. A strong correlation between cytokines production was found but groups differed in this aspect. CONCLUSION: By multiplex analysis, it may be possible, for example, to define the risk immunological response pattern among relatives or to monitor the immune response in patients on immune modulation therapy.