[The assessment of T regulatory cells in the peripheral blood of children with metabolic syndrome]. / Ocena limfocytów T regulatorowych we krwi obwodowej dzieci z zespolem metabolicznym.

UNLABELLED: The metabolic syndrome is defined as the co-existence of risk factors for the development of cardio-vascular disease: obesity, hypertension, insulin resistance and dyslipidemia. For a few years there has been a growing interest in immune-inflammatory aspects of obesity and metabolic syndrome. According to many authors the disturbances in the number and(or) function of T regulatory cells are responsible for autoimmune diseases. It is possible that they play a role in a pathogenesis of chronic inflammation accompanying obesity. THE AIM OF THE STUDY was to determine the percentages of T regulatory cells in children with metabolic syndrome. MATERIAL AND METHODS: Fourty seven children with metabolic syndrome were prospectively enrolled into the study according to the IDF criteria (central obesity and two of: hypertension, hypertriglycerydemia, low HDL, hyperglycemia/glucose intolerance/diabetes). With the use of five-colour flow cytometry the following percentages of T cells in the peripheral blood were assessed: CD4(+), CD4(+)CD25(high), CD4(+)CD25(high)FoxP3(+), CD4(+)CD25(high)CD127(low), CD4(+)CD25(high)FoxP3(+)CD127(low). RESULTS: In the group of children with metabolic syndrome we noted lower percentages of CD4(+)CD25(high) cells compared to control children: 1.7 vs. 3.7% (p = 0.01). The differences in CD4(+)CD127(low) cells were not statistically significant: 15.7 vs. 17.6% (p = 0.1). We did not observe the differences between examined and control group in the percentages of CD4(+)CD25(high)CD127(low) and CD4(+)CD25(high)FoxP3(+) cells (respectively: 0.54% vs. 0.58%, 0.49 vs. 0.59%, p > 0.05). CONCLUSIONS: Results of our investigation suggest the lower percentages of CD4(+)CD25(high) but not other Tregs subpopulation cells in children with metabolic syndrome. The further research concerning the role of Tregs in the pathogenesis of immunologic disturbances accompanying metabolic syndrome will continue.

[Relationship between bone mineral content and growth disorders in children with juvenile idiopathic arthritis]. / Zaleznosc miedzy masa mineralna kosci a zaburzeniami wzrastania u dzieci chorych na mlodziencze idiopatyczne zapalenie stawów.

UNLABELLED: In chronic inflammatory processes in children efforts are made to evaluate bone mineral content (BMC) with the use of densitometric parameters, which at the same time determine equivalent of lean body mass (LBM). This functional analysis of the musculoskeletal system by using DXA method seems to be particularly useful for the examination of bone mass in children suffering from juvenile idiopathic arthritis (JIA). THE AIM OF THE STUDY was to assess body mass content having regard of the BMC/LBM ratio in JIA children, depending on the degree of growth inhibition, disease advancement phase and the therapy applied. MATERIAL AND METHODS: The study comprised 97 children aged 5-18 (mean age 12.7+/-3.8 years), 45 girls and 52 boys with diagnosed JIA according to ILAR criteria of 1997. The average duration of disease was 4.1+/-3.1 years. Antropometric and densitometric measurement was made in every child. Body height was defined by Standard Deviation Height Velocity Score - SDHVS (SDS). Patients were divided into 2 groups: I - 28-group with SDS ratio < -2.0; 11 - 69 children with normal height. For the evaluation of disease development and advancement of anatomic changes in joints criteria of Steinbrocker were applied: I grade - without joint damage, II - insignificant or moderate joints damage, III-IV - established deformation. The densitometric research was conducted with the use of double-energy X-ray absorptiometry (DXA) method. Bone mineral density (BMD) was assessed in the whole skeleton (TB BMD), in the vertebras L2-L4 (SB MD), Z-score index for SBMD and BMC, LBM, TB BMC/LBM defined by the Z-score index and compared to norms for age and growth. RESULTS: Bone mineral density defined as Z-score index for SBMD <-2.0 was reported in 21 children (21.6%). Muscle-skeletal Z-score index for TB BMC/LBM in relation to norms of gender and growth lower than -1.0 was proved in 23 children (23.6%). Considerably lower Z-score index for TB BMC/LBM (p < 0.01) characterized children with growth inhibition and children with significant joints damage (p < 0.02). There was no significant correlation between densitometric parameters and applied treatment with glucocorticoids and without glucocorticoids. CONCLUSIONS: Decrease of bone mineral mass defined as muscle-skeletal Z-score index for TB BMC/LBM was found in almost quarter of patients within the group of JIA children. In the group of children with growth deficiency and with larger joints damage bone mass was significantly lower.

Bone mineral metabolism in children with juvenile idiopathic arthritis--preliminary report.

INTRODUCTION: There are very few reports assessing bone mineral mass and its metabolism in the course of juvenile idiopathic arthritis (JIA). STUDY OBJECTIVE: To assess the levels of selected serum markers of bone formation (OCN) and resorption (CTx) in JIA children. MATERIAL AND METHODS: The study involved 52 children with JIA diagnosed according to the EULAR criteria of 1997, aged 6-18 years. All patients underwent densitometric measurements using dual-energy X-ray absorptiometry (DXA) to assess TBBMD (g/cm2), Spine BMD (g/cm2), Z-score for SBMD, TBBMC (g), and LBM (g). The following parameters were determined in blood serum: the level of osteocalcin (OCN) and C-terminal type I alpha-collagen chain telopeptide (CTx) using the Elecsys 2010 system (N-MID Osteocalcin, Beta-CrossLaps). A gender- and age-matched control group consisted of 16 healthy children. RESULTS: The mean concentrations of both osteocalcin (p<0.001) and CTx (p<0.005) were significantly higher in JIA patients as compared to the healthy controls (OCN 113.2+/-54.9 ng/ml vs. 70.2+/-48.3 ng/ml; CTx 1.4+/-0.5 mug/l vs. 1.2 +/-0.45 microg/l). The concentrations of the bone turnover markers were significantly reduced in children with higher degrees of joint destruction compared to those with anatomically normal joints (p<0.05). The mean concentration of CTx showed a significant negative correlation with the TB BMC/LBM Z-score (p<0.05). Reduced bone mass (Z-score for SBMD< -2.0) was found in 23.6% of the affected children. CONCLUSIONS: The JIA patients had elevated levels of OCN and CTx compared to the healthy controls. Reduced bone turnover was observed in children with higher degrees of joint destruction.

Elevated matrix metalloproteinase 9 and tissue inhibitor of metalloproteinase 1 in obese children and adolescents.

Matrix metalloproteinases (MMPs) have been implicated in the atherosclerotic process and risk factors for the disease such as hypertension, hyperlipidemia, or diabetes mellitus in adults. So far, circulating levels of MMPs and their tissue inhibitors (TIMPs) have not been assessed in children and adolescents with obesity, a known risk factor for cardiovascular disease. Plasma levels of MMP-9 and TIMP-1 were measured immunoenzymatically in 45 obese children and adolescents, aged 15 +/- 1.8 years. The control group consisted of 28 healthy children, aged 14.5 +/- 2.5 years. MMP-9 and TIMP-1 concentrations were higher in obese children than in the control group (MMP-9: 553.5 +/- 311 vs 400.4 +/- 204 ng/mL, respectively; P = .02; TIMP-1: 161.2 +/- 32 vs 143.1 +/- 20.1 ng/mL, respectively; P = .03). We found significantly higher levels of MMP-9 in obese children with coexisting hypertension than in obese normotensive patients (635 +/- 308 vs 450 +/- 289 ng/mL, respectively; P = .04). MMP-9 correlated with body mass index (BMI) (r = 0.33, P = .005) and fasting insulin (r = 0.3, P = .013); TIMP-1 correlated with BMI (r = 0.35, P = .006). In the group of obese hypertensive children (n = 25), MMP-9 correlated with BMI (r = 0.41, P = .001), systolic blood pressure (r = 0.41, P = .002), fasting insulin (r = 0.37, P = .006), and homeostasis model assessment index of insulin resistance (r = 0.27, P = .03). TIMP-1 correlated with BMI (r = 0.33, P = .025) and systolic (r = 0.38, P = .008) and diastolic (r = 0.47, P = .001) blood pressure. In the regression models, MMP-9 was found to be dependent on fasting insulin (R(2) = 0.16, P = .04), and TIMP-1 on BMI (R(2) = 0.14, P = .04). In the obese hypertensive group, TIMP-1 was dependent on diastolic blood pressure (R(2) = 0.18, P = .04). Obese children and adolescents have elevated plasma concentrations of MMP-9 and TIMP-1. Coexistence of hypertension may exacerbate alterations of extracellular matrix turnover in these patients. It might be hypothesized that elevated MMP and TIMP concentrations may be related to increased cardiovascular risk in obese and particularly in obese hypertensive children and adolescents.

Relationship between endothelial dysfunction, carotid artery intima media thickness and circulating markers of vascular inflammation in obese hypertensive children and adolescents.

BACKGROUND: Adhesion molecules released by dysfunctional endothelium are considered as markers of vascular inflammation in early atherosclerosis. Non-invasive ultrasound methods are now available to detect first preclinical signs of the disease. AIM: To investigate the relationship between selected adhesion molecules and ultrasound indicators of early atherosclerosis: endothelial function measured by flow-mediated dilatation (FMD) and intima media thickness (IMT). PATIENTS: The study group consisted of 85 children, mean age 14.6 years, of whom 22 were obese, 31 were hypertensive, and 32 obese and hypertensive. The control group included 26 healthy children. METHODS: Adhesin concentrations were determined by ELISA. FMD and IMT were evaluated by ultrasound. RESULTS: A positive correlation was found between sICAM-1 (soluble intercellular adhesion molecule 1) and IMT (r = 0.32, p = 0.013, 95% CI: 0.11 to 0.49) and a negative correlation between IMT and FMD (r = -0.26, p = 0.04, 95% CI: -0.43 to -0.04) in the whole study group. In the particular groups, we found significant correlations only in obese hypertensive children. sICAM-1 correlated positively with IMT (r = 0.52, p = 0.001, 95% CI: 0.2 to 0.72) and negatively with FMD (r = -0.31, p = 0.027, 95% CI: -0.6 to -0.2). sE-selectin correlated positively with IMT (r = 0.41, p = 0.012). In regression models, IMT correlated with sICAM-1 (beta = 0.37, p = 0.03) and body mass index (beta = 0.55, p = 0.02), and FMD correlated negatively with sICAM-1 (beta = -0.47, p = 0.04). CONCLUSIONS: The association between inflammatory markers of the endothelium with impaired vasodilatation activity and the first atherosclerotic structural changes in the common carotid arteries were found in obese hypertensive children and adolescents. The coexistence of obesity and hypertension predisposes these young patients to closely related disturbances connected with early atherosclerosis.

[Early atherosclerosis changes in children with juvenile idiopathic arthritis]. / Wczesne zmiany miazdzycowe u dzieci chorych na mlodziencze idiopatyczne zapalenie stawów.

UNLABELLED: Pathogenesis of the two diseases: juvenile idiopathic arthritis (JIA) and atherosclerosis are similar-they include increased inflammation markers, homocysteine and lipids levels, inflammatory cytokines. It seems to be important to look for new diagnostic methods to find atherosclerosis in early stage- ultrasonography of the carotid artery with intima-media thickness (IMT) measurement and serum homocysteine level. The aim of this study was assessment of preclinical markers of atherosclerosis--concentration of homocysteine and IMT MATERIAL AND METHODS: The study group consists of 40 children with JIA (20 girls and 20 boys) aged 4-16 years; 32 children with oligoarthritis JIA and 8--polyarthritis JIA. Control group consists of 23 healthy children aged 3-17 years. We investigated serum levels of homocysteine, CRP, lipids and also IMT of the carotid artery. Children with JIA had statistically significant increase of IMT compare to control group (0.43 mm vs. 0.40 mm) and higher IMT in polyarthritis group compare to oligoarthritis children (0.46 mm vs. 0.43 mm). We also find statistically significant correlation between IMT and disease duration (p = 0.003, r = 0.45). RESULTS: Children with JIA had also statistically significant increase of homocysteine level compare to control group (8.2 micromol/l vs. 6.05 micromol/l) and higher homocysteine level in polyarthritis group compare to oligoarthritis children (8.58 micromol/l vs. 7.88 micromol/l). We also find statistically significant correlation between IMT and homocysteine level (p = 0.02, r = 0.36). CONCLUSIONS: There is accelerated atherosclerosis in JIA children that's why we need to find new methods to evaluate it in very early stage so we could help these patients to prevent its complication such as heart disease, stroke etc in the future.

[Analysis of Fas, FasL and Caspase-8 expression in thyroid gland in young patients with immune and non-immune thyroid diseases]. / Analiza ekspresji czasteczek Fas, FasL oraz kaspazy 8 w tkance gruczolu tarczowego u mlodych pacjentów z chorobami immunologicznymi i nieimmunologicznymi gruczolu tarczowego.

INTRODUCTION: Apoptosis, programmed cell death is a regulating mechanism enabling the removal of superabundantly produced and unnecessary at the certain moment cells. Disturbances of the apoptosis regulation contribute to the pathogenesis of many diseases, including autoimmune thyroid disorders. The aim of this study was to estimate expression of proapoptotic Fas/FasL and caspase-8 in thyroid tissues in patients with Graves' disease (GD), non-toxic nodular goiter (NTNG) and Hashimoto's thyroiditis (HT). MATERIAL AND METHODS: Inclusion criteria of Graves' patients were: large goiter, ophthalmopathy, TRAb > 5 U/L, positive titre of anti-TPO and anti-TG antibodies and concentration of TSH < 0.45 microIU/mL for more the 2-3 months from an onset of the disease. Isolated thyrocytes were identified by indirect method: in the first stage mouse monoclonal antibodies (mAbs) anti-TPO were bound to rabbit anti-mouse antibodies IgG (Fab')2 labeled FITC. To obtained cellular suspension mAbs directed against apoptotic Fas/FasL molecules labeled with PE (Phycoerythrin) was added. All investigations were performed on Coulter EPICS XL flow cytometer. Detection of apoptotic proteins was confirmed by Western Blot and immunohistochemistry methods using mAbs in DAB chromogene visuality and marked by Mayer's haematoxylin. Evaluation of caspase-8 expression in thyroid follicular cells was performed by Western Blot test. RESULTS: The analysis of Fas and FasL expression on surface of thyroid follicular cells was higher in patients with Hashimoto's thyroiditis (38%, 26%) in comparison with patients with Graves' disease (18%, 14%). In case of patients with Hashimoto's thyroiditis significantly lower percentage of thyroid tissue infiltrating immune Fas+ (13%) and FasL+ (22%) T cells in comparison with Graves' patients (33%, 43% respectively) was observed . Identification of proapoptotic Fas and FasL molecules in the thyroid follicular cells revealed higher expression of both proteins in patients with GD (++,++) and HT (+++; +++, respectively) in comparison with NTNG patients (+/0; +/0). Caspase-8 expression was detected in band 55 kDa using Western Blot test in patients with thyroid autoimmune diseases. CONCLUSIONS: We conclude that alteration in the expression of proapoptotic proteins in thyroid follicular cells may play a role in pathogenesis of thyroid autoimmune disorders. In addition, suppression of apoptosis in Graves' disease led to increased proliferation of thyroid follicular cells which is responsible for goiter formation.

[Evaluation of central corneal thickness in children and adolescents with type I diabetes mellitus]. / Analiza centralnej grubosci rogówki u dzieci mlodziezy z cukrzyca insulinozalezna.

PURPOSE: To evaluate corneal thickness in diabetic and nondiabetic patients. MATERIALS AND METHODS: The central corneal thickness (CCT) was investigated in 100 eyes of 100 patients with diabetes type I and in 99 nondiabetic patients (99 eyes). The mean diabetic patients age was 15.31 +/- 3.18 years. The mean age in control group was 14.3 +/- 2.2 years. Corneal thickness was measured by non-contact microscope Topcon SP-2000P Statistical analysis was performed to assess systemic factors (patient age, sex, duration of diabetes mellitus, hemoglobin A1c value, diabetes control) related to CCT. SAS STAT (Release 8.2) program, the independent t-test, Kolmogorow-Smirnow test and Bartlett test were used to compare differences between the diabetic and control group. RESULTS: In our study the mean CCT in diabetic eyes was 0.54 +/- 0.03 mm and was significantly increased, compared to control group (0.525 +/- 0.037 mm). None of systemic factors was correlated with CCT. CONCLUSIONS: Our findings indicate that diabetes mellitus affects thickness of cornea in adolescents. Evaluation of endothelium in specular microscope should be performed in the diabetic patients.

[Correlation analysis between diabetic retinopathy and early atherosclerotic changes in adolescents with type 1 diabetes -- preliminary report]. / Zwiazek miedzy rozwojem retinopatii cukrzycowej a wystepowaniem wczesnych zmian miazdzycowych u mlodziezy z cukrzyca typu 1 -- doniesienie wstepne.

OBJECTIVE: Recent studies have shown a correlation between advanced diabetic retinopathy and late stages of atherosclerosis. There are no findings on a possible relation between diabetic retinopathy and diseases of the cardiovascular system at their earliest stage in young people with diabetes type 1. The purpose of the study was to analyze a correlation between diabetic retinopathy and early atherosclerotic changes in adolescents with type 1 diabetes. RESEARCH DESIGN AND METHODS: The study included 28 adolescents aged 17.6+/-1.4 years suffering from type 1 diabetes mellitus for 7.9+/-3.1 years, the mean age of the disease onset - 9.5+/-3.7 years, a mean level of HbA1c - 8.6+/-1.9%. Eight patients with developing simple retinopathy, were separated from the whole group of young people. First control group consisted of the remaining patients with type 1 diabetes chosen with regard to age and sex, without disease complications. Second control group consisted of 11 healthy young people. The function of endothelium by measuring the brachial artery dilatation -- FMD and the intima-media complex thickness of the common carotid arteries were evaluated ultrasonographically. RESULTS: Young people with retinopathy had higher systolic pressure: 133+/-19 mmHg in comparison with patients without complications: 117+/-14 mmHg (p<0.05) and healthy people: 115+/-8 mmHg (p<0.05). All patients with diabetes showed significantly lower FMD (7.6+/-5.1%, p<0.05). In the group with retinopathy, FMD equaled 7.8+/-4.1% (p=0.04) and in the group without retinopathy - 7.6+/-5.5% (p<0.05) in comparison with 12.1+/-5.1% in healthy volunteers. Significantly higher IMT was found in all patients with diabetes in comparison with healthy young people: 0.49+/-0.06 vs. 0.42+/-0.03 mm (p<0.001). Patients with retinopathy had a significantly higher value of IMT in comparison not only with controls but also with patients without complications: 0.56+/-0.06 vs. 0.47+/-0.03 mm (p<0.001). CONCLUSIONS: 1. Young people with type 1 diabetes had a significantly impaired function of endothelium and higher IMT in comparison with healthy young people. 2. Adolescents with retinopathy were characterized by significantly higher values of systolic arterial blood pressure when compared to patients without complications 3. Higher IMT was found in patients with diabetic retinopathy in comparison with patients without complications, which may suggest that macrovascular changes are more advanced in case of complications than in patients without retinopathy.

[Serum levels of matrix metaloproteinase 2 (MMP-2), matrix metaloproteinase 9 (MMP-9) and tissue inhibitor of metaloproteinase 2 (TIMP-2) in children and adolescents with growth hormone deficiency]. / Ocena stezenia metaloproteinazy macierzy pozakomorkowej 2 (MMP-2), metaloproteinazy macierzy pozakomorkowej 9 (MMP-9) i tkankowego inhibitora metaloproteinazy 2 (TIMP-2) u dzieci i mlodziezy z niedoborem hormonu wzrostu.

INTRODUCTION: Matrix metaloproteinases (MMPs) have been implicated in various pathological processes including inflammatory response, atherosclerosis and cardiovascular disease. Growth hormone deficiency (GHD) is associated with prematury atherosclerosis and cardiovascular disease. Circulating levels of matrix metaloproteinases and their tissue inhibitors (TIMPs) so far have not been assessed in children and adolescents with GHD. MATERIAL AND METHODS: Serum levels of matrix metaloproteinase 2 (MMP-2), matrix metaloproteinase 9 (MMP-9) and tissue inhibitor of metaloproteinase 2 (TIMP-2) were measured in 44 (11 girls and 33 boys) children and adolescents with newly diagnosed GHD [age (mean+/-SD) 12.5+/-2.7 years, height 1.3+/-0.1 m, body surface area (BSA) 1.1+/-0.2 m(2) and body mass index (BMI) 17.4+/-2.2 kg/m(2)] and in 32 (11 girls and 21 boys) healthy children and adolescents (age 12.4+/-2.9 years, height 1.6+/-0.2 m, BSA 1.4+/-0.3 m(2) and BMI 18.7+/-2.6 kg/m(2)). Human MMP-2, MMP-9 and TIMP-2 measurements were carried out with the use of ELISA kits. RESULTS: Patients with GHD had significantly higher concentrations of MMP-2 (287.2+/-60.5 vs. 235.8+/-41.3 ng/ml, p<0.0001) and TIMP-2 (81.4+/-14.9 vs. 62.7+/-15.9 ng/ml, p<0.0001) levels than the control healthy group. There was no difference in MMP-9 levels (338.5+/-197.9 vs. 276.3+/-121.7 ng/ml, p=0.12) between patients with GHD and controls. CONCLUSIONS: Children and adolescents with GHD have elevated serum concentrations of MMPs-2 and TIMP-2.

[Selected matrix metalloproteinases (MMP-2, MMP-9) in obese children and adolescents]. / Ocena stezenia wybranych metaloproteinaz macierzy zewnatrzkomórkowej (MMP-2, MMP-9) u dzieci i mlodziezy z otyloscia prosta.

INTRODUCTION: In the last years it has been proved that matrix metalloproteinases participate in the development of all stages of atherosclerotic process. It has been suggested that plasma levels of metalloproteinases can be a novel, inflammatory marker of atherosclerosis. THE AIM OF THE STUDY was to evaluate plasma levels of selected matrix metalloproteinases (MMP-2 and MMP-9) in obese children and adolescents. MATERIAL AND METHODS: We studied 45 children and adolescents with simple obesity aged 15+/-1.8 years. The control group comprised 28 healthy, slim children aged 14.5 years. The obese children were studied according to coexistence of hypertension, hyperlipidemia, positive family history of cardiovascular diseases and insulin resistance. Levels of matrix metalloproteinases were assessed by use of ready ELISA kits (R&D Systems). RESULTS: MMP-9 level in the study group equaled 553.5+/-311 ng/ml and was significantly higher compared to the control group: 400.4+/-204 ng/ml, p=0.02. MMP-2 level in the study group was significantly lower compared to the control group: 211+/-37 ng/ml vs. 258+/-56 ng/ml, p<0,0001. In obese children with coexisting hypertension we found higher levels of MMP-9 when compared to children with obesity only: 635+/-308 ng/ml vs. 450+/-289 ng/ml (p=0,04), MMP-2 levels remained similar in both groups. CONCLUSIONS: Different levels of plasma matrix metalloproteinases in obese and hypertensive children compared to healthy, slim controls can indicate an altered metabolism of the extracellular matrix (ECM) of vessels and heart muscle. Changed metabolism of ECM may be of significant importance in enhancing the atherosclerotic process in this young patients.

[Metalloproteinases MMP-2 and MMP-9 and their inhibitors TIMP-1 and TIMP-2 levels in children and adolescents with type 1 diabetes]. / Aktywnosc metaloproteinaz MMP-2 i MMP-9 oraz ich inhibitorów TIMP-1 i TIMP-2 u dzieci i mlodziezy z cukrzyca typu 1.

INTRODUCTION: Abnormal activation of the matrix metalloproteinases (MMPs) in diabetes mellitus leads to extracellular matrix changes through the structural protein composition changes. The metalloproteinases inhibitors (TIMPs) are regulatory factors in this activity. Not all regulating mechanisms are completely known, especially in patients with type 1 diabetes. THE AIM OF STUDY: evaluation of MMP-2, MMP-9 and TIMP-1, TIMP-2 levels in children and adolescents with type 1 diabetes. MATERIAL AND METHODS: 74 patients in the mean age 15 years (+/-3.0) suffering from type 1 diabetes for mean 6.6 years (+/-3.6) took part in the study. Patients were treated with flexible multiple daily insulin (n=54) and with continuous subcutaneous insulin infusion - personal insulin pump (n=20). MMP-2, MMP-9, TIMP-1 and TIMP-2 blood serum levels were measured in all patients. 45 healthy persons matched for age, without atherosclerosis risk factors, with proper BMI and lipids levels were in the control group. RESULTS: MMP-2 level as well as TIMP-2 and TIMP-2 levels were significantly higher in patients with type 1 diabetes in comparison to the control group (p respectively <0,01; <0,02; <0,001). We observed higher MMP-9 level in obese patients than in patients with BMI value below 90 pc for sex and age (p<0,02). We noted lower MMP-2 level in patients with chronic complications and/or arterial hypertension (n=24) in comparison to patients without that kind of complications (p<0,05). Positive correlation between TIMP-1 level and HbA1c level was noted. Age of patients as well as BMI negatively correlated with MMP-2 and TIMP-2 and positively with TIMP-1. We observed a correlation between MMP-2, TIMP-2 and especially TIMP-1 with lipid levels. Strong positive correlation was noted between MMP-2 and TIMP-2 (r=0.8; p<0,0001). CONCLUSIONS: 1. MMP-2, TIMP-1 and TIMP-2 levels are higher in patients with type 1 diabetes than in the control group. 2. Age, diabetes duration, metabolic control, BMI and lipids levels have influence on the MMPs/TIMPs system. 3. TIMP-1 is supposed to be the key marker of metabolic disturbances in type 1 diabetes.

[Role of apoptosis in autoimmune thyroid disorders]. / Rola apoptozy w chorobach autoimmunologicznych tarczycy.

Apoptosis, programmed cell death, is a physiological phenomenon, necessary for normal function of every organism. This is an active process, per-current with a participation of the cellular metabolism embracing the activation of genes and the synthesis of proteins. The signal to apoptosis can be started practically in every cell of our organism. Disturbances of the apoptosis regulation determine the essential link of the pathogenesis of many diseases, including autoimmune thyroid disorders. Such molecules as FasL, TNF (tumor necrosis factor), TRAIL (the ligand inducing apoptosis), inducing different apoptotic pathway can play the key-role in the pathogenesis of Graves' disease or Hashimoto's thyroiditis. Besides in the destructive thyroiditis an important role is also played by proteins from the bcl-2 family and the proinflammatory cytokines. The aim of this publication is to present the influence of different factors on the apoptosis and the role of programmed cells death in autoimmune thyroid diseases.

[Composition of linoleic and arachidonic acids in phosphatidylcholine of erythrocytes membrane and IL-6 and TNF-alpha in serum and C-reactive protein concentration in children with juvenile idiopathic arthritis]. / Zawartosc kwasu linolowego i arachidonowego w fosfatydylocholinie blon erytrocytarnych oraz IL-6 i TNF-alpha w surowicy a stezenie bialka C-reaktywnego u dzieci chorych na mlodziencze idiopatyczne zapalenie stawów.

UNLABELLED: The study objective was to determine the relationship, if any, between the levels of arachidonic and linoleic acids in erythrocyte membrane phosphatidylcholine (PCH), serum interleukin 6 (IL-6) and TNFalpha levels and C-reactive protein (CRP) in juvenile idiopathic arthritis (JIA). MATERIALS AND METHODS: The study involved 49 children with JIA, aged 3-18 years (mean 11.3 +/- 3.9), and 29 healthy subjects. The JIA children were divided into 2 groups: group I--24 children (in exacerbation period) with mean CRP level of 15.6 mg/L +/- 13.3 and group II--25 children (in remission period--joint swelling-free) with CRP of 7.8 mg/L +/- 5.8 on average. Lipids were extracted according to a modified Folch's method. Fatty acids in erythrocyte membrane PCH were identified using gas chromatography (Hewlett-Packard 5890). The levels of IL-6 and TNFalpha were determined by ELISA, using Quantikine sets: R&D System (USA), while CRP was measured by nephelometric method on a BN II apparatus (Behring). RESULTS: We found a significant decrease in the level of linoleic acid (p < 0.05) and a statistically insignificant reduction in arachidonic acid in JIA patients as compared to the controls. The decrease in linoleic acid was more pronounced in the active phase of JIA (p < 0.001. The higher serum CRP level was accompanied by a significantly elevated level of IL 6 (p < 0.05). The concentration of TNFa was elevated, but the difference had no statistical significance. CONCLUSIONS: The levels of linoleic and arachidonic acids in erythrocyte PCH decreased and the concentrations of IL-6 and TNF-alpha increased in JIA children in the active phase of the disease. The differences intensified with a rise in CRP.

[The assessment of 24-hour ambulatory blood pressure monitoring (ABPM), microalbuminuria and diabetic autonomous neuropathy in children with type 1 diabetes and hypertension]. / Ocena 24-godzinnego monitorowania cisnienia tetniczego, mikroalbuminurii oraz autonomicznej neuropatii cukrzycowej u dzieci z cukrzyca typu 1 i nadcisnieniem tetniczym.

INTRODUCTION: The late complications of diabetes consisted of autonomic neuropathy, nephropathy, which more often coexist with hypertension in children with type 1 diabetes mellitus. THE AIM OF THE STUDY was to assess the connections between changes in the autonomous nervous system, 24-hour ABPM and daily albumin excrection in children with hypertension and type 1 diabetes mellitus. MATERIAL: The group consisted of 72 patients with diabetes (diabetes duration time 6.5+/-1.5 years). 34 patients of that group have hypertension. The control group consisted of 30 healthy children matched according to age and sex. RESULTS: In children with hypertension we found significantly often occurrence of microalbuminuria (13/34 i 1/38, p<0.001). In 17 patients from the group with hypertension and 17 patients without hypertension we affirm signs of autonomic neuropathy. The values of heart rate variability (HRV) were significantly decreased in the group with hypertension as compared to the control group. A stepwise multiple regression analysis with hypertension as a dependent variable and diabetes duration time, microalbuminuria, HbA1c level, HRV parameters and a presence of autonomous neuropathy as predictors proved that hypertension is associated with higher HbA1c level (b=0.35), the presence of autonomous neuropathy (b=0.28), and lower HF values (b=0.41) (p<0.01). CONCLUSIONS: Hypertension in children with type 1 diabetes mellitus is correlated with the presence of autonomous neuropathy, higher HbA1c level and lowered values of heart rate variability parameters.

[Ultrasonographic evaluation of selected parameters of the endothelial function in brachial arteries and IMT measurements in carotid arteries in children with diabetes type 1 using personal insulin pumps--preliminary report]. / Ultrasonograficzna ocena wplywu zastosowania osobistych pomp insulinowych na stopien zaawansowania procesu miazdzycowego w tetnicach szyjnych wspólnych i ramiennych u dzieci z cukrzyca typu 1--doniesienie wstepne.

INTRODUCTION: Type 1 diabetes is a known risk factor for arterial atherosclerosis. The first symptoms can be found even in childhood. The ultrasonographic measurements of intimal plus medial thickness in carotid arteries (IMT) and flow mediated dilatation (FMD) evaluated in brachial arteries, play a known role in the detection in these cases. The diabetes treatment intensification is an important factor in delaying early atherosclerotic changes. Currently, intensive treatment of children's diabetes with use of continuous subcutaneous insulin infusion with personal insulin pumps is gaining more and more popularity. THE AIM OF THIS STUDY was the evaluation of IMT and FMD indexes in children suffering from type 1 diabetes in the context of treatment intensification (multidose insulin injections v. personal insulin pumps). MATERIAL AND METHODS: We examined 64 children (29 boys and 35 girls) in the mean age 15.5 years treated with the multidose insulin injections method and 10 children using personal insulin pumps (4 girls and 6 boys) in the mean age 14.5 years. Using high resolution ultrasonography we evaluated IMT values in carotid arteries and FMD parameters in brachial arteries. In our analysis we estimated the blood concentration of lipid parameters, values of systolic and diastolic blood pressure, the age of diabetes onset, duration time of the illness and the values of HbA1c as a marker of metabolic control. RESULTS: We noticed significantly higher FMD values in patients treated with personal insulin pumps (13.7 vs. 5.5%, p=0.001). IMT values were similar in both groups (0.52 vs. 0.5 mm, p=0. 41). The level of HDL cholesterol was higher and triglycerides lower in the group with treatment intensification. The metabolic control was the same in both groups. In patients treated by the multidose insulin injections IMT correlated with systolic blood pressure values. We didn't notice any correlation between IMT and FMD in any group. CONCLUSIONS: 1. Treatment intensification (personal insulin pumps) influences better vascular endothelial function in type 1 diabetic children and seems to be a significant tool in delaying the atherosclerotic process. 2. We need more examinations to explain the role of treatment intensification in common carotid arteries wall morphology in type 1 diabetic children. 3. The ultrasonographic detection of atherosclerotic changes in arterial vessels can help in the evaluation of the changes due to different methods of diabetes treatment.

[Bone mineral density in children and adolescents with idiopathic chronic musculoskeletal pain syndromes]. / Masa kostna u dzieci i mlodziezy z idiopatycznymi bólami miesniowo-szkieletowymi.

PURPOSE: There is a limited information regarding associations between bone mineral density (BMD) and idiopathic musculoskeletal pain in pediatric subjects. The aim of this cross-sectional study was to assess whether children and adolescents with pain syndrome have lower BMD than healthy subjects. MATERIAL AND METHODS: Eighty-nine subjects (49 girls, 40 boys) aged 5-18 years with chronic non-rheumatic musculoskeletal pain and daily calcium consumption below 500 mg were involved in the study. The subjects were divided in three groups: I--prepubertal (5-9 years), II--pubertal (9-15 years) and III--adolescents (15-18 years). Bone mineral density in the total skeleton (total BMD) and lumbar spine (spine BMD) was examined using dual energy x-ray absorptiometry (DXA) and compared to reference data. Serum calcium, ionized calcium, phosphate, alkaline phosphatase and its bone-fraction were estimated. RESULTS: Low BMD (below the 5th percentile) was found in about 50% of the participants with pain syndromes in each measurement site. A significantly decreased Spine BMD was observed in those children who reported pain symptoms in the spine region and lower limbs (Z-score for Spine BMD was = -1.55; -1.41) compared to subjects with non-localized pain. The lowest mean Z-score (-1.85) for Spine BMD was found in the postpubertal group (III) compared with groups I and II. CONCLUSIONS: As one-half of children and adolescents reporting musculoskeletal pain had low BMD it is possible that the symptoms are associated with an inadequate bone mass accrual during growth. Pain localized in the spine region and in lower extremities may be a selective and site-specific symptom of juvenile osteoporosis without fractures. The chronic musculoskeletal pain suggests a rationale for bone density testing during growth.

[Cytofluorometric analysis of chosen markers of apoptosis CD95/CD95L (Fas/FasL) in thyroid tissues from young patients with Graves' disease and Hashimoto's thyroiditis]. / Cytofluorymetryczna analiza wybranych markerów apoptozy Fas/FasL (CD95/CD95L) w tkance gruczolu tarczowego u mlodocianych pacjentów z choroba Gravesa-Basedowa oraz zapaleniem tarczycy typu Hashimoto.

BACKGROUND: Apoptosis, one of the forms of programmed cell death, is a physiologic process of cell death that is central to normal development and occurs in response to a variety of physiologic and pathophysiologic stimuli. In the thyroid, abnormal apoptotic activity may be involved in a variety of diseases such as Hashimoto thyroiditis and Graves disease. The aim of this study was to estimate the expression of chosen apoptotic molecules CD95 (Fas) and CD95L (FasL) on the surface of thyroid follicular cells in application of mouse monoclonal antibodies #64 which recognized B antigen regions of thyroid peroxidase (TPO) and infiltrating inflammatory cells. MATERIAL AND METHODS: The investigation was performed on thyroid cells isolated from surgically treated thyroid tissues of 15 patients with Graves' disease (GD), 15 patients with a nontoxic multinodular goiter (NTMG) and 15 aspirates obtained by FNAB from patients with Hashimoto thyroiditis (HT). The thyrocytes were identified by an indirect method: in the first stage we added mouse monoclonal autoantibodies specific for TPO (mAb #64) regions and in the second stage we conjugated this complex with rabbit anti-mouse antibodies IgG (Fab')2 with FITC. In the next step the cellular suspension was completed with suitably well-chosen two-colour monoclonal antibodies marked (PE or PerCP) (Becton Dickinson) directed against suitable apoptotic (Fas/FasL) molecules. All investigations were performed by flow cytometry using Coulter EPICS XL apparatus. RESULTS: The percentages of thyroid cells were estimated with expression of region B antigenic TPO in reference to individual apoptotic molecules. The analysis of Fas and FasL expression in thyroid tissues revealed significantly increased percentage of intrathyroidal T cells with CD95+ (p<0.005, p<0.001), CD95L+ (p<0.02, p<0.01) and both CD95/CD95L (ns, p<0.05) expression in comparison to percentages of T cells in patients with HT and NTMG. In addition, on the surface of thyroid follicular cells in patients with GD (p<0.01, p<0.01) and NTMG (p<0.001, p<0.004) we observed a lower percentage of thyrocytes with CD95 and CD95L molecules than in cases with HT. The expression of both apoptotic molecules on thyroid cells was higher (18%) in patients with HT in comparison to the percentages of positive cells in patients with GD (p<0.02, p<0.002) and NTMG, 8% and 1%, respectively. CONCLUSIONS: We conclude that alterations in the expression of death receptors and their ligands on the surface of thyroid follicular cells may play a role in the regulation of apoptosis in thyroid autoimmune disorders.

Soluble adhesion molecules (sICAM-1, sVCAM-1) and selectins (sE selectin, sP selectin, sL selectin) levels in children and adolescents with obesity, hypertension, and diabetes.

The attachment of monocytes and lymphocytes to endothelial cells, which initiates atherosclerosis, arises under the influence of adhesion molecules. The preclinical phase of this disease lasts many decades, and this provides an opportunity for the presymptomatic detection of high-risk subjects. We evaluated levels of the adhesion molecules: sICAM-1 (soluble intercellular adhesion molecule 1), sVCAM-1 (soluble vascular adhesion molecule 1), sE selectin, sP selectin, and sL selectin in children with atherosclerosis risk factors (n = 123, mean age 15.1 years) (obese [n = 17], hypertensive [n = 25], obese with hypertension [n = 30], type 1 diabetic [n = 51]). Twenty-seven healthy children formed the control group, mean age 15.2 years. sICAM-1 was higher in the study group compared with control (314.1 +/- 61 vs 264.9 +/- 55 ng/mL, P < .01). The same was found for sVCAM-1 (513.7 +/- 187 vs 407.9 +/- 76 ng/mL, P < .05) and E selectin (86.04 +/- 33.6 vs 62.1 +/- 20.3 ng/mL, P < .01). sP-selectin and sL-selectin levels were not different compared with controls. E selectin correlated with body mass index (BMI; r = 0.18, P = .03), total cholesterol (r = 0.2, P = .016), and triglycerides (r = 0.22, P = .008). sICAM-1 correlated with BMI (r = 0.19, P = .019) and systolic blood pressure (r = 0.13, P = .045). In multiple linear regression analysis, sE selectin was found to be associated with triglycerides (R2 = 0.29, P = .045), sICAM-1 dependent on BMI (R2 = 0.58, P = .047), and sVCAM-1 dependent on total cholesterol (R2 = 0.51, P = .006). Elevated concentrations of sICAM-1, sVCAM-1, and E selectin were found in obese, hypertensive, and diabetic children. We conclude that endothelial activation appears in these children, and adhesion molecules are related to the earliest stages of atherosclerosis.

Analysis of costimulatory molecules OX40/4-1BB (CD134/CD137) detection on chosen mononuclear cells in children and adolescents with Graves' disease during methimazole therapy.

Antibody synthesis follows interactions between the T cell receptor (TCR) on activated T lymphocytes and the main histocompatibility complex (MHC) present on APC cells, resulting in lymphocyte proliferation, as well as cytokine synthesis and release. The involvement of costimulatory markers OX40/4-1BB/4-1BBL leads to the enhancement of signals which are necessary for lymphocyte activation in addition to the antigen-specific signal and may prevent anergy. The aim of this study was to estimate the expression of OX40 and 4-1BB molecules on peripheral blood cells in patients with Graves' disease (GD) (n = 35, mean age 16.5 +/- 6.1 years) and non-toxic nodular goiter (NTNG) (n = 35, mean age 16.2 +/- 4.7 years), in comparison with sex- and age-matched healthy controls (n = 35, mean age 16.2 +/- 2.1 years). Expression of the costimulatory molecules on mononuclear cells was analyzed by three-color flow cytometry using a Coulter EPICS XL cytometer. Stimulating and blocking antibodies to the TSH-receptor using JPO9 CHO cells in unfractionated serum were measured by a highly sensitive commercial radioimmunoassay. The analysis of OX40/4-1BB expression in patients with newly recognized Graves' disease revealed a statistically significant increase in the percentage of CD134+ T cells (7% vs 1.4%, p <0.001) and CD137+ T cells (3.2% vs 0.8%, p <0.04) compared to the control group. After 2-6 months of methimazole therapy, the percentage of these cells in the peripheral blood of hyperthyroid patients returned to normal values. In addition, the expression of 4-1BBL (CD137L) was detected only on the surface of active monocytes in patients with untreated GD (3.8%), while in the group with nodular goiter and controls the values were trace (0.6% and 0.2%, respectively). We conclude that the changes of expression of costimulatory molecules on the surface of peripheral blood T cells and their significant relationship with the level of antithyroid antibodies indicate an involvement of these molecules in the pathogenesis of Graves' disease. A marked increase in the percentage of CD134/ CD137+ T cells at disease onset may indicate the need for more aggressive therapy in Graves' disease and for a greater duration than the standard 3-year period.