Dense deposit disease in an adolescent male mimicking acute post-streptococcal glomerulonephritis.

BACKGROUND: Dense deposit disease (DDD), a subtype of complement factor 3 glomerulopathy (C3G), is a rare entity associated with dysregulation of the alternative complement pathway. It usually affects children, with a 50% likelihood of progression to end-stage renal disease within ten years of diagnosis. Description of the case: We report the case of an adolescent male with acute nephritic syndrome and nephrotic range proteinuria, initially diagnosed as acute post-streptococcal glomerulonephritis (APSGN). Despite his spontaneous improvement, renal biopsy, performed due to a persistently low C3 level for over 18 weeks, confirmed the diagnosis of DDD. Complement and genetic studies showed high levels of C3-nephritic factor and risk polymorphisms for developing the disease. He was treated with prednisolone and mycophenolate mofetil (MMF). At the last follow-up, 15 months from onset, the serum creatinine level and 24h-hour total protein excretion were normal. CONCLUSION: C3G (including the DDD subtype) should be suspected in apparent APSGN with atypical clinical features at presentation/follow-up, even in the case of spontaneous improvement. Timely and accurate diagnosis, based on histopathological, complement, and genetic studies, is important to initiate the appropriate treatment aimed at preventing or slowing the disease progression. HIPPOKRATIA 2020, 24(4): 191-193.

Association of Low Ficolin-Lectin Pathway Parameters with Cardiac Syndrome X.

In patients with typical angina pectoris, inducible myocardial ischaemia and macroscopically normal coronaries (cardiac syndrome X (CSX)), a significantly elevated plasma level of terminal complement complex (TCC), the common end product of complement activation, has been observed without accompanying activation of the classical or the alternative pathways. Therefore, our aim was to clarify the role of the ficolin-lectin pathway in CSX. Eighteen patients with CSX, 37 stable angina patients with significant coronary stenosis (CHD) and 54 healthy volunteers (HC) were enrolled. Serum levels of ficolin-2 and ficolin-3, ficolin-3/MASP-2 complex and ficolin-3-mediated TCC deposition (FCN3-TCC) were determined. Plasma level of TCC was significantly higher in the CSX than in the HC or CHD group (5.45 versus 1.30 versus 2.04 AU/ml, P < 0.001). Serum levels of ficolin-2 and ficolin-3 were significantly lower in the CSX compared to the HC or CHD group (3.60 versus 5.80 or 5.20 µg/ml, P < 0.05; 17.80 versus 24.10 or 26.80 µg/ml, P < 0.05). The ficolin-3/MASP-2 complex was significantly lower in the CSX group compared to the HC group (92.90 versus 144.90 AU/ml, P = 0.006). FCN3-TCC deposition was significantly lower in the CSX group compared to the HC and CHD groups (67.8% versus 143.3% or 159.7%, P < 0.05). In the CSX group, a significant correlation was found between TCC and FCN3-TCC level (r = 0.507, P = 0.032) and between ficolin-3/MASP-2 complex level and FCN3-TCC deposition (r = 0.651, P = 0.003). In conclusion, in patients with typical angina and myocardial ischaemia despite macroscopically normal coronary arteries, low levels of several lectin pathway parameters were observed, indicating complement activation and consumption. Complement activation through the ficolin-lectin pathway might play a role in the complex pathomechanism of CSX.

Complement activation and its prognostic role in post-cardiac arrest patients.

Cardiac arrest causes generalized ischaemia/hypoxia, and subsequent resuscitation inflicts reperfusion injury, the pathology of which is not fully understood. Moreover, predicting the prognosis of comatose, post-cardiac arrest patients is a complex clinical challenge. We hypothesized that the extent of complement activation might be a reliable predictor of mortality in this population. Forty-six comatose cardiac arrest patients were enrolled into our prospective cohort study, conducted in a tertiary care university clinic. All subjects were cooled to 32-34 °C body temperature for 24 h and then allowed to rewarm to normothermia. All patients underwent diagnostic coronary angiography. On admission, at 6 and 24 h, blood samples were taken from the arterial catheter. In these, complement products (C3a, C3, C4d, C4, SC5b9 and Bb) were measured by ELISA in blood samples. Patients were followed up for 30 days; 22 patients (47.8%) died by the end of this period. We observed that complement activation (determined as the C3a to C3 ratio) was higher in non-survivors than in survivors at each time point. In the multivariate Cox regression analysis, the C3a/C3 ratio determined 24 h after the initiation of therapeutic hypothermia predicted 30-day mortality regardless of age, sex and the APACHE II score. Complement activation occurs in post-cardiac arrest patients, and its extent correlates with 30-day survival. The C3a/C3 ratio might prove useful for estimating the prognosis of comatose post-cardiac arrest patients.

The level of complement C3 is associated with the severity of atherosclerosis but not with arterial calcification in peripheral artery disease.

AIM: Recent evidences show correlations between atherosclerosis and the serum level of third component of complement (C3). However, there is less data on the connection of C3 and the severity of atherosclerosis. The aim of our study was to evaluate the association of serum C3 levels with atherosclerosis and arterial calcification in patients with chronic lower extremity atherosclerosis. METHODS: In a single centre cross-sectional study 103 patients and 109 healthy controls were examined. Sera were analyzed for C3. To identify the severity of atherosclerosis and calcification, ankle-brachial Doppler index, angiographic Bollinger score, arterial calcification score and carotid intima-media thickness was determined. RESULTS: Serum level of C3 was significantly higher in the lower extremity atherosclerosis group than in healthy patients (P=0.00004). In the patient group, serum level of C3, C4 was significantly and inversely associated with ABI (r=-0.246, P=0.014), (r=-0.259, P=0.011). C3 inversely correlates with Bollinger score (r= -0.357, P=0.028). Among our patients no correlation was found between C3 levels and CS (P=0.672, r=-0.046) or between C3 levels and carotid IMT (r=0.104, P=0.351). The serum levels of different complement components were associated with C-reactive protein, Hba1c, peptide-C and insulin. CONCLUSION: Our results suggest that C3 serum levels are associated with ABI and angiographic parameters of atherosclerosis, but do not relate to the severity of calcification.

Evidence-based hydro- and balneotherapy in Hungary--a systematic review and meta-analysis.

Balneotherapy is appreciated as a traditional treatment modality in medicine. Hungary is rich in thermal mineral waters. Balneotherapy has been in extensive use for centuries and its effects have been studied in detail. Here, we present a systematic review and meta-analysis of clinical trials conducted with Hungarian thermal mineral waters, the findings of which have been published by Hungarian authors in English. The 122 studies identified in different databases include 18 clinical trials. Five of these evaluated the effect of hydro- and balneotherapy on chronic low back pain, four on osteoarthritis of the knee, and two on osteoarthritis of the hand. One of the remaining seven trials evaluated balneotherapy in chronic inflammatory pelvic diseases, while six studies explored its effect on various laboratory parameters. Out of the 18 studies, 9 met the predefined criteria for meta-analysis. The results confirmed the beneficial effect of balneotherapy on pain with weight bearing and at rest in patients with degenerative joint and spinal diseases. A similar effect has been found in chronic pelvic inflammatory disease. The review also revealed that balneotherapy has some beneficial effects on antioxidant status, and on metabolic and inflammatory parameters. Based on the results, we conclude that balneotherapy with Hungarian thermal-mineral waters is an effective remedy for lower back pain, as well as for knee and hand osteoarthritis.

Serum concentration of immunoglobulin G-type antibodies against the whole Epstein-Barr nuclear antigen 1 and its aa35-58 or aa398-404 fragments in the sera of patients with systemic lupus erythematosus and multiple sclerosis.

Several studies suggest that infection by Epstein-Barr virus (EBV) might be one of the environmental factors which facilitates the development of autoimmune disorders in genetically susceptible individuals. Recent data indicate that high anti-Epstein-Barr nuclear antigen 1 (EBNA)-1 immunoglobulin (Ig)G titre is a strong risk factor for multiple sclerosis (MS) in patients both with and without the main genetic predisposing trait, human leucocyte antigen (HLA)-DRB1*15:01. Because no similar studies have been published in systemic lupus erythematosus (SLE) patients, we determined the HLA-DRB1*15:01 carrier state and the serum titres against the whole EBNA-1 and its small fragments aa35-58 and aa398-404 in 301 SLE patients, 135 MS patients and in 345 healthy controls. The carrier state of the HLA-DRB1*15:01 allele was deduced from genotyping of a tagSNP (rs3135388) by applying a Taqman-based assay. The serum concentrations of antibodies to EBNA-1 and its aa35-58 or aa398-404 fragments were determined using a commercial assay (ETI-EBNA-G) and home-made enzyme-linked immunosorbent assays, respectively. The serum concentration of anti-EBNA-1 antibodies was significantly (P < 0·001) higher both in MS and SLE patients than in controls. Similar significant differences were found both in HLA-DRB1*15:01 carriers and non-carriers. Furthermore, titres of antibodies against the aa35-58 EBNA-1 fragment were elevated both in MS and SLE patients. By contrast, the levels of aa398-404 EBNA-1 antibodies were elevated significantly only in the SLE patients. These findings indicate that high anti-EBNA-1 IgG titres are HLA-DRB1*15:01-independent risk factors not only for MS, but also for SLE, while high antibody titres against the aa398-404 fragment are characteristic for SLE.

Circulating ficolin-2 and ficolin-3 in normal pregnancy and pre-eclampsia.

Ficolins are soluble molecules of the innate immune system that recognize carbohydrate molecules on microbial pathogens, apoptotic and necrotic cells. They act through two distinct routes: initiating the lectin pathway of complement activation and mediating a primitive opsonophagocytosis. In this study, we measured plasma levels of ficolin-2 and ficolin-3 in 60 pre-eclamptic patients, 60 healthy pregnant women and 59 healthy non-pregnant women by enzyme-linked immunosorbent assay (ELISA). Circulating levels of complement activation products (C4d, C3a, SC5b9), angiogenic factors (soluble fms-like tyrosine kinase-1, placental growth factor) and markers of endothelial activation (von Willebrand factor antigen), endothelial injury (fibronectin) and trophoblast debris (cell-free fetal DNA) were also determined. Plasma levels of ficolin-2 were significantly lower in healthy pregnant than in healthy non-pregnant women, while ficolin-3 levels did not differ significantly between the two groups. Furthermore, pre-eclamptic patients had significantly lower ficolin-2 and ficolin-3 concentrations than healthy non-pregnant and pregnant women. In the pre-eclamptic group, plasma ficolin-2 levels showed a significant positive correlation with serum placental growth factor (PlGF) concentrations and significant inverse correlations with serum levels of soluble fms-like tyrosine kinase-1 (sFlt-1), blood urea nitrogen and creatinine, serum lactate dehydrogenase activities, as well as with plasma VWF:antigen, fibronectin and cell-free fetal DNA concentrations. In conclusion, circulating levels of ficolin-2 are decreased in the third trimester of normal pregnancy. There is a further decrease in plasma ficolin-2 concentrations in pre-eclampsia, which might contribute to the development of the maternal syndrome of the disease through impaired removal of the trophoblast-derived material released into the maternal circulation by the hypoxic and oxidatively stressed pre-eclamptic placenta.

Complement activation in thrombotic thrombocytopenic purpura.

BACKGROUND: Ultra-large von Willebrand factor and deficiency of its cleaving protease are important factors in the events leading to thrombotic microangiopathy; however, the mechanisms involved are only partly understood. Whereas pathological activation of the alternative complement pathway is linked to atypical hemolytic uremic syndrome, the role of complement activation in thrombotic thrombocytopenic purpura (TTP) is unknown. The aim of this study was to investigate whether signs of complement activation are characteristic of TTP. PATIENTS AND METHODS: Twenty-three patients with TTP (18 women, median age 38 years) and 17 healthy controls (13 women, median age 38 years) were included. Complement parameters (C3, Factors H, I, B and total alternative pathway activity) together with complement activation fragments (C3a) or complexes (C1rs-INH, C3bBbP, sC5b9) were measured by ELISA or RID. ADAMTS13 activity and anti-ADAMTS13 inhibitory antibodies were measured by the VWF-FRET73 assay. RESULTS: Increased levels of C3a, and SC5b9 were observed in TTP during acute episodes, as compared with healthy controls. Decreased complement C3 levels indicative of complement consumption occurred in 15% of acute TTP patients. Significant decrease of complement activation products C3a and SC5b9 was observed during plasma exchange (PEX). The sustained presence of anti-ADAMTS13 inhibitory antibodies in complete remission was associated with increased complement activation. CONCLUSION: These data document in an observational study the presence of complement activation in TTP. Further investigation is needed to determine its potential pathogenetic significance.

OS004. A link between the complement system and angiogenic imbalance inpreeclampsia: ficolin-2 deficiency.

INTRODUCTION: An excessive maternal systemic inflammatory response to pregnancy, as well as an imbalance between circulating angiogenic factors and their antagonists plays a central role in the pathogenesis of preeclampsia. The complement system, as part of innate immunity, is fundamental to the host's immune defense against microbial pathogens, apoptotic and necrotic cells. Both of its excessive activation and deficiencies can lead to various disorders. OBJECTIVES: The aim of this study was to determine circulating levels of components of the complement system and their relationship to those of angiogenic factors in normal pregnancy and preeclampsia. METHODS: Sixty preeclamptic patients, 60 healthy pregnant women and 59 healthy non-pregnant women were involved in this case-control study. Circulating levels of C1rC1sC1-inh, C3bBbP, C4d, C3a, SC5b9, ficolin-2, ficolin-3, soluble fms-like tyrosine kinase-1 (sFlt-1) and placental growth factor (PlGF), as well as activity of the complex of mannose-binding lectin and mannose-binding lectin-associated serine protease 2 (MBL-MASP2 complex) were measured. For statistical analyses, non-parametric methods were applied. RESULTS: Circulating levels of C3bBbP, C4d, C3a, SC5b9, sFlt-1, PlGF, as well as MBL-MASP2 activity were significantly higher, while ficolin-2 concentrations were significantly lower in healthy pregnant than in healthy non-pregnant women. Furthermore, preeclamptic patients had significantly higher C1rC1sC1-inh, C3bBbP, C4d, C3a, SC5b9 and sFlt-1 levels and significantly lower ficolin-2, ficolin-3 and PlGF concentrations than healthy pregnant women. In the groups of healthy pregnant women and preeclamptic patients, plasma ficolin-2 levels showed a significant positive correlation with serum PlGF concentrations and a significant inverse correlation with serum levels of sFlt-1. There was no other relationship between complement components and angiogenic factors in either study group. CONCLUSION: Elevated levels of activation products in the systemic circulation indicate complement activation with increased terminal complex formation in preeclampsia, which seems to be independent from alterations in circulating angiogenic factors. Nevertheless, low ficolin-2 concentrations might contribute to the angiogenic imbalance in preeclampsia by impaired removal of the sFlt-1-containing trophoblast-derived material released into the maternal circulation by the hypoxic and oxidatively stressed preeclamptic placenta.

Serum fetuin-A levels inversely correlate with the severity of arterial calcification in patients with chronic lower extremity atherosclerosis without renal disease.

AIM: Fetuin-A is a hepatic glycoprotein that inhibits extraosseous calcification. Lower serum fetuin-A concentration was associated with severe arterial calcification in patients with end stage renal disease. We evaluated the association of serum fetuin-A levels and the severity of atherosclerosis in patients with peripheral vascular disease having normal renal function. METHODS: In this cross-sectional study among 93 chronic atherosclerotic patients with lower extremity vascular disease, systemic atherosclerosis and calcification was assessed by ultrasound (carotid intima-media thickness/IMT/, calcification at the abdominal aorta, carotid and femoral bifurcations, aortic and mitral valves) and angiography (Bollinger score). Standard serum markers of inflammation, diabetes, renal function, ankle-brachial indexes and traditional risk factors for atherosclerosis were noted and Fontaine classification was applied for the severity of symptoms. RESULTS: The patients mean (SD) age was 59.95 (7.61) years, 78% were men, 35% had diabetes. Serum fetuin-A level showed significant negative correlation with ultrasound calcification score (P=0.018, r=-0.257) and Bollinger angiographic score (P=0.035, r=-0.347). Fetuin-A did not correlate with IMT or Fontaine classification. Fetuin-A also showed significant correlation with albumin, transferrin and hemoglobin A1c (r=0.287, 0.305 and 0.219, respectively at P<0.05). Logistic regression analysis confirmed the association between fetuin-A and calcification score (OR: 3.03, CI: 1.05-8.7), P=0.039) independent of traditional risk factors. CONCLUSION: Our data show that serum fetuin-A levels inversely correlate with the severity of atherosclerosis in nonuremic patients with symptomatic chronic lower limb ischemia. These data support a putative protective role for fetuin-A in the development of arterial calcification.

Diagnosis and classification of hemolytic uremic syndrome: the Hungarian experience.

BACKGROUND: Hemolytic uremic syndrome (HUS) is a rare disease with various etiologies, making the identification of the specific forms and appropriate treatment difficult. Therefore, clinical and laboratory data from these patients need to be analyzed in national and international registries. Herein we have described 47 Hungarian HUS patients with detailed laboratory and clinical data obtained between 2008 and 2010. METHODS: Blood samples and clinical data of 47 patients with HUS diagnosed according to characteristic clinical signs were submitted for diagnostic evaluation, including complement protein and genetic analysis, measurement of ADAMTS13 activity and antibody analysis against O157LPS and factor H. RESULTS: There were 8 patients with typical diarrhea-positive HUS; 13 with atypical HUS (aHUS) and 26 with secondary HUS/thrombotic thrombocytopenic purpura group characterized by signs of complement consumption and decreased ADAMTS13 activity. Thus, decreased total alternative pathway activity is a promising diagnostic parameter with good sensitivity for aHUS. CONCLUSIONS: These observations highlight the requirement for multiple diagnostic tests together with clinical data to identify the specific cause of HUS. Because the long-term prognosis of aHUS, eg, graft survival after renal transplantation, may vary according to the molecular etiology, it is important for all affected patients to undergo a detailed molecular diagnosis of the disease. There is a clear clinical need for the development and application of novel assay in this field to allow more rapid efficient diagnosis of patients who undergo a first episode of HUS.

Human anti-60 kD heat shock protein autoantibodies are characterized by basic features of natural autoantibodies.

Anti-human Hsp60 autoantibodies--known risk factor of atherosclerosis--were investigated in a mouse model and in samples of healthy subjects: polyreactivity, presence in cord blood samples of healthy newborns and life-long stability were tested. In IgM hybridoma panel from mouse spleens, polyreactivity of anti-Hsp60 autoantibodies was studied. In healthy pregnant women, umbilical vein and maternal blood samples were collected after childbirth, anti-Hsp-60 and -65 IgM and IgG levels were measured. Life-long stability of anti-Hsp-60 levels was studied on healthy patients during 5 years. ELISA was used in all studies. Polyreactivity of IgM clones of newborn mice and lifelong stability of these autoantibodies in healthy adults were established. IgM anti-Hsp60 autoantibodies in cord blood of healthy human infants were present, however, there was no correlation between maternal and cord blood IgM anti-Hsp60 concentrations. It is proposed that presence of anti-Hsp60 autoantibodies--as part of the natural autoantibody repertoire--may be an inherited trait. Level of anti-Hsp60 autoantibodies may be an independent, innate risk factor of atherosclerosis for the adulthood.

Adrenomedullin and endothelin-1 are related to inflammation in chronic heart failure.

BACKGROUND: Adrenomedullin (ADM) and endothelin-1 (ET-1) are novel promising peptide biomarkers in chronic heart failure (CHF). According to recent studies among their pleiotropic effect they play roles in the regulation of inflammation. The aim of the study was to measure the above mentioned two vasoactive peptides in parallel in a well characterized population of patients with CHF, and study their associations with inflammatory markers. MATERIALS AND METHODS: A total of 186 patients (138 male, 48 female) with <45% left ventricular ejection fraction (LVEF), and without acute inflammatory disease, were enrolled. Plasma midregional-proADM (MR-proADM) and C-terminal-proET-1 (CT-proET-1) were determined by a novel sandwich immunoluminometric assay. RESULTS: Increased MR-proADM and CT-proET-1 plasma levels were measured in patients with severe CHF (NYHA III-IV) as compared to the group of NYHA I-II (p<0.0001). MR-proADM and CT-proET-1 levels showed significant negative correlation with serum albumin and prealbumin levels (p

Inflammatory- and immune responses in relation to bacterial replication in mice following re-infections with Chlamydophila pneumoniae.

OBJECTIVE: Investigation of chronic infections with Chlamydophila pneumoniae. METHODS: BALB/c mice were repeatedly infected with C. pneumoniae and tested during a 1-year period. Production of histamine, IFN-gamma, IL-6 and antibodies was monitored by ELISA. Live bacteria were cultured and DNA was detected by PCR. Cellular immunity was tested by ELISPOT. RESULTS: After re-infections, culture positivity and persistence of DNA in lungs and blood were shorter. Detection of DNA at late time points indicated persistent infection in a few mice. Histamine was produced after primary and re-infections, and the level correlated with the number of viable bacteria in lung. IFN-gamma, IL-6 levels, IgG2/IgG1 ratio, IgA titres, and level of chlamydial heat-shock protein antibodies were higher after re-infections. IgM antibodies were demonstrated even after re-infections. High number of IFN-gamma-producing splenocytes was observed after the third inoculation. CONCLUSION: These results promote an understanding of the patho- and immune mechanisms after C. pneumoniae re-infections.

Effects of vaccination with heat shock proteins on streptozotocin induced diabetes in histidine decarboxylase knockout mice.

RATIONALE: Type 1 diabetes mellitus (T1D) is an immune mediated disease in which heat shock proteins (hsps) may be involved in the development of the disease. Furthermore, vaccination with different hsps prevented the development of multiple low-dose streptozotocin (STZ) induced autoimmune diabetes in C57BL/KSJ mice. Histamine influences many aspects of the immune response, including Th1/Th2 balance and antibody production. Therefore, a study of diabetes-related immune processes was considered of interest in histidine decarboxylase knockout (HDC-KO) mice. AIM OF THE STUDY: The aim of our study was i) to characterize antibody production in response to vaccination with p277 or hsp65 in wild type (WT) BALB/c and HDC-KO mice, and ii) to establish a possible correlation between vaccination and the changes in the pattern of STZ diabetes. MATERIALS AND METHODS: An ELISA was employed to measure the hsp65- and p277-specific antibody levels. To induce diabetes multiple low-dose of STZ was used. RESULTS: Vaccination with p277 and hsp65 altered the pattern of STZ diabetes both in HDC-KO and WT animals, characterized by a transient increase followed by sustained reduction of blood sugar levels as compared to controls. However, vaccination with hsp65 and p277 caused a significant anti-p277 and anti-hsp65 antibody level increase only in WT animals. CONCLUSION: Multiple low-doses of STZ were able to induce diabetes in HDC-KO mice and the development of diabetes was prevented by vaccination with hsps. This protection developed in spite of the fact that vaccination caused a significant antibody level increase only in WT animals. To explain the therapeutic effect of vaccination we need further examination of the HDC KO strain.

Smoking and a complement gene polymorphism interact in promoting cardiovascular disease morbidity and mortality.

We have demonstrated previously that carriers of a genotype called C4B*Q0 (silent allele of the C4B gene) have a substantially increased risk to suffer from myocardial infarction or stroke, and are selected out from the healthy elderly population. Because smoking carries a major risk for cardiovascular disease (CVD), it seemed worthwhile to study if these two factors interact. Study 1 involved 74 patients with angina pectoris (AP), 85 patients with recent acute myocardial infarction (AMI) and 112 survivors of a previous AMI and 382 controls from Iceland. Study 2 involved 233 patients with severe CVD and 274 controls from Hungary. Smoking habits were registered for each subject. The number of C4A and C4B genes was determined by phenotyping or genotyping. Compared to controls, C4B*Q0 carrier frequency was significantly higher at diagnosis in Icelandic smokers with AP (P = 0.005) and AMI (P = 0.0003) and Hungarian smokers with severe coronary artery disease (P = 0.023), while no such difference was observed in non-smoking subjects. Age-associated decrease in C4B*Q0 observed previously in two remote Caucasian populations was found, in the present study, to be associated strongly with smoking, and to already occur in smokers after age 50 years both in Iceland and Hungary. Our findings indicate that the C4B*Q0 genotype can be considered as a major covariate of smoking in precipitating the risk for AMI and associated deaths.

Association of elevated serum heat-shock protein 70 concentration with transient hypertension of pregnancy, preeclampsia and superimposed preeclampsia: a case-control study.

Our aim was to investigate the association between serum heat-shock protein (Hsp) 70 concentration and hypertensive disorders of pregnancy. One hundred and forty-two pregnant women with hypertensive disorders (93 with preeclampsia, 29 with transient hypertension of pregnancy and 20 with superimposed preeclampsia) and 127 normotensive, healthy pregnant women were included in the study. Serum Hsp70 concentration was measured using enzyme-linked immunosorbent assay. The serum Hsp70 concentration was significantly higher in patients with transient hypertension of pregnancy, in preeclamptic patients and in patients with superimposed preeclampsia than in the control group (median (25-75 percentile): 0.66 (0.52-0.84), 0.55 (0.42-0.80), 0.61 (0.42-0.91) ng/ml vs 0.31 (0.27-0.39) ng/ml, respectively; P<0.001). Multivariate logistic regression analysis showed independent association of elevated serum Hsp70 level with transient hypertension of pregnancy, preeclampsia and superimposed preeclampsia. The difference in serum Hsp70 concentration between preeclamptic patients and the control group was statistically significant in each gestational age category. In the groups of preeclamptic and superimposed preeclamptic patients, there was no significant difference in serum Hsp70 concentration between mild and severe preeclamptic patients, between patients with late and early onset of the disease, as well as between preeclamptic patients without and with foetal growth restriction. In conclusion, serum Hsp70 concentration is elevated in transient hypertension of pregnancy, in preeclampsia and in superimposed preeclampsia. Circulating Hsp70 may not only be a marker for these conditions, but might also play a role in their pathogenesis. However, further studies are needed to explore its role in the pathogenesis of hypertensive disorders of pregnancy.

Th1 and Th2 cell responses of type 1 diabetes patients and healthy controls to human heat-shock protein 60 peptides AA437-460 and AA394-408.

RATIONALE: Type 1 diabetes mellitus (T1) is considered to be an immune mediated disease. Based on previous findings it might be suggested that heat shock protein 60 (Hsp60) could be involved in the mediation of the development of the disease. Furthermore a bias toward Th1 immune response was observed in T1D patients where the level of Th1 cytokines was elevated, while the level of Th2 was decreased. AIM OF THE STUDY: To determine Th1 (IFN-gamma) and Th2 (IL-13) cytokine levels in T1 diabetic and control subjects as well as to determine whether there is a shift towards Th1 or Th2 immune response. MATERIALS AND METHODS: ELISPOT (Enzyme-linked ImmunoSPOT) analysis was employed to differentiate antigen specific T-cell responses of a Th1 (IFN-gamma) or Th2 (IL-13) type. 11 T1 diabetic patients and 9 healthy controls were investigated. For T-cell stimulation, we used a polyclonal mitogen or Tetanus toxoid (TT) as positive controls and two peptide antigens Hsp60 AA394-408 and Hsp60 AA437-460. RESULTS: In case of Hsp60 AA437-460 we found significantly decreased Th2 response in patients, although there was no significant difference in Th1 response. In case of Hsp60 AA394-408 and positive controls there was no significant difference. CONCLUSION: Comparing the control and diabetic subjects a significant shift towards Th1 response in T1 diabetes mellitus for Hsp60 AA437-460 was observed.

Changes in the plasma concentration of soluble thrombomodulin in patients with severe carotid artery stenosis after eversion endarterectomy.

OBJECTIVE AND DESIGN: The purpose of the study was to investigate the putative role of soluble thrombomodulin (sTM) in severe carotid artery stenosis. MATERIALS AND METHODS: We prospectively studied 64 patients who were undergoing carotid endarterectomy (2001-2003). Plasma sTM concentration was determined in each patient before surgery and at 14 months postsurgery. -308 TNF-alpha promoter polymorphism was also determined. RESULTS: Strong negative correlation was found between the preoperative duplex scan values and the plasma sTM concentrations (R = -0.418, p = 0.0006). Patients with 308 A TNF-alpha genotype had significantly lower (p = 0.0415) preoperative sTM values than their counterparts with no such polymorphism. Soluble TM concentrations measured in plasma samples taken at the end of the postsurgical follow-up period of 14 months duration were significantly higher compared to the preoperative values (p < 0.0001). CONCLUSIONS: Our present findings indicate that sTM may be adsorbed to the atherosclerotic plaques or inflamed endothelium in carotid arteries. The pathological significance of this adsorption remains to be determined.

High levels of C-reactive protein with low total cholesterol concentrations additively predict all-cause mortality in patients with coronary artery disease.

BACKGROUND: This study aimed to investigate independent and additive predictive effects of raised C-reactive protein (CRP) levels and decreased total cholesterol levels on mortality in patients with chronic coronary artery disease (CAD). Low total cholesterol (TC) levels are associated with worsened survival in chronic and acute diseases. Elevated CRP level is an important predictor of vascular events and mortality in patients with CAD. Potential inhibition of immune activation by circulating lipoproteins could be a link between cholesterol and inflammatory markers. MATERIALS AND METHODS: A group of 387 patients (median age 59 years) with CAD and with or without severe heart failure (HF) were followed for a median of 5.06 years. Serum total cholesterol and CRP concentrations were measured at enrollment. RESULTS: The relationship between lipoproteins, CRP and survival was explored. High CRP concentrations were in significant association with severity of HF and predicted worsened survival in patients with CAD (hazard ratio 5.214, 95% CI 1.762-15.427). The association between CRP levels and mortality was independent of potential confounding factors such as age, body-mass index, severity of HF, smoking habits, hypertension and TC levels. The prediction of mortality by low TC levels was significant (hazard ratio 2.932, 95% CI 1.021-8.422). Furthermore, patients with increased CRP and decreased TC (additive predictive effect) phenotype had 11.714-times higher risk (95% CI 2.619-52.385) of being nonsurvivors than patients with low CRP/high TC. CONCLUSIONS: High CRP levels and low TC concentrations are independent and additive predictors of mortality in patients with CAD. Our data indicate that joint analysis of circulating lipoproteins and inflammatory biomarkers may improve prediction of survival in patients with CAD.