Relationship of pregnancy-associated plasma protein-a to renal function and dialysis modalities.

BACKGROUND: The aim of the study was to determine pregnancy-associated plasma protein-A (PAPP-A), which was recently described as a new marker of cardiovascular events, in patients with chronic renal insufficiency/failure and to find out its relationship to renal function and to prominent markers of oxidative stress (advanced oxidation protein products--AOPP) and inflammation (C-reactive protein--CRP). METHODS: The studied group consisted of 36 chronic hemodialysis patients (HD), 10 patients treated with continuous ambulatory peritoneal dialysis (CAPD) and 38 patients with chronic renal insufficiency (CHRI) not yet dialyzed. PAPP-A was measured by Time Resolved Amplified Cryptate Emission technology. Determination of AOPP is based on a spectrophotometric method. RESULTS: PAPP-A levels are statistically significantly elevated in the both groups of dialyzed patients in comparison with healthy subjects (27.0 +/- 16.5 mIU/l in HD and 14.07 +/- 6.73 mIU/l in CAPD vs. 8.22 +/- 2.7 mIU/l in the control group, p < 0.0001 and p < 0.001, respectively, p < 0.05 HD vs. CAPD). The mean serum PAPP-A levels in the CHRI patients not yet dialyzed were not significantly higher in comparison with the control group (9.72 +/-4.44 vs. 8.22 +/- 2.7 mIU/l, n.s.). In the CHRI not dialyzed patients, we found a significant positive correlation between serum creatinine and PAPP-A levels (r = 0.68, p < 0.05). In comparison with controls, AOPP and CRP levels were significantly higher in HD patients [AOPP 155.0 +/- 37.9 micromol/l, p < 0.0001 vs. controls, CRP 10.0 (4.6- 26.9) mg/l (median, interquartile range), p < 0.0001 vs. controls], CAPD patients [AOPP 118.5 +/- 25.8 micromol/l, p < 0.0001 vs. controls, CRP 7.7 (2.0-18.8) mg/l, p < 0.01 vs. controls] and AOPP levels in chronic renal failure patients not yet dialyzed (98.5 +/- 43.24 micromol/l, p < 0.01 vs. controls). The correlations between PAPP-A and AOPP (r = 0.49, p < 0.05) and PAPP-A and CRP (r = 0.48, p < 0.05) serum concentration were statistically significant in HD patients. In CAPD patients, neither a correlation between PAPP-A and AOPP nor a correlation between PAPP-A and CRP were found. CONCLUSION: We can conclude that serum PAPP-A levels sensitively reflect the changes in renal function, depend on dialysis modality, and may represent a novel marker associated with inflammation and oxidative stress in chronic renal failure patients.

[Levels of advanced oxidation protein products (AOPP) in the first trimester of pregnancy]. / Hladiny produktu pokrocilé oxidace proteinu (AOPP) v prvním trimestru tehotenství.

Advanced oxidation protein products (AOPP) may be sensitive biomarkers for protein damage mediated by reactive oxygen species. AOPP were measured in the serum of 41 pregnant women in the 8th-12th week of pregnancy. Parameters of prenatal screening in the first trimester (pregnancy-associated plasma protein A--PAPP-A and free beta human chorionic gonadotrophin--free beta HCG) and anticardiolipin antibodies (ACA) IgG and IgM were determined as well. A group of healthy blood donors--women and men was used for comparison. AOPP were determined spectrophotometrically according to Witko-Sarsat [24] (absorbance at 340 nm) and were expressed in chloramine units (mumol/l). Other analytes were determined by immunoanalytic methods. AOPP levels in pregnant women in the first trimester are significantly higher in comparison with blood donors--women (89.46 +/- 33.38 mumol/l vs 57.34 +/- 16.31 mumol/l, p < 0.0001) but there is no statistically significant difference between pregnant women and blood donors--men (89.46 +/- 33.38 mumol/l vs 78.60 +/- 44.01 mumol/l). AOPP level does not correlate either with the age of pregnant women or with the parameters of prenatal screening and ACA IgG and IgM. Higher levels of AOPP in the serum of pregnant women in comparison with women--blood donors may reflect an increase of oxidative stress in pregnancy.

Antibodies against beta 2-glycoprotein I (beta 2-GP I) and their relationship to fetoplacental antigens.

Binding of beta 2-GP I to anionic phospholipids is thought to be the major antigen required in the reaction of anticardiolipin antibodies to phospholipids. The aim of this study was to investigate the changes of anti-beta 2-GP I IgG during the first and second trimester of pregnancy and the relationship between the levels of anti-beta 2-GP I and fetoplacental antigens and the correlation between anti-beta 2-GP I IgG and antibodies against oxidized low-density lipoprotein IgG (oLAb) in serum of pregnant women. We determined anticardiolipin antibodies (ACA) IgG and maternal serum levels of alpha 1-fetoprotein (AFP), human chorionic gonadotrophin (HCG) and trophoblast-specific beta 1-glycoprotein (SP1) in 204 pregnant women in the first and second trimester. From this group we selected 52 serum samples positive for ACA IgG and 16 samples negative for ACA IgG. In the samples of selected patients, the levels of anti-beta 2-GP I IgG and oLAb IgG were determined. Anti-beta 2-GP I IgG levels significantly decreased in the second trimester (6.2+/-9.3 U/ml, mean +/- S.D.) in comparison with the first trimester (8.3+/-10.4 U/ml) (p=0.05). Multiple of median (MoM) AFP correlated negatively but not significantly in the first trimester with anti-beta 2-GP I (r = -0.261, p = 0.12). In the second trimester this correlation was significantly negative (r = -0.278, p = 0.04). The Spearman correlation coefficients for MoM HCG and anti-beta 2-GP I were 0.158 for the first trimester and 0.174 for the second trimester. MoM SP1 also did not correlate significantly with anti-beta 2-GP I in both trimesters. The correlation between anti-beta 2-GP I IgG and oLAb IgG was not significant (r = -0.06). In the first trimester 40 % serum samples were positive for anti-beta 2-GP I IgG and negative for oLAb IgG or vice versa, while 60 % samples in the second trimester were positive only for one determined autoantibody. We can conclude that the levels of anti-beta 2-GP I IgG decrease during the second trimester probably as the result of the effects of some immunosuppressive agents associated with pregnancy. The finding of negative correlation between AFP and anti-beta 2-GP I suggests that anti-beta 2-GP I has an influence on fetus development.

Antibodies against oxidized low density lipoproteins in pregnant women.

Oxidized low density lipoproteins (oxLDL) formed in vivo induce a humoral immune response. Oxidative modification of LDL renders it immunogenic and a heterogeneous population of specific anti-oxLDL antibodies is produced. These antibodies could represent a biological marker of oxidative stress and serve as markers of atherosclerosis. Autoantibodies against oxLDL (oLAb) have been detected in human subjects practically of every age. oLAb also appear in the blood of pregnant women. Some studies have shown that the levels of antibodies to oxLDL were elevated in women with established preeclampsia. The present study was aimed to estimate the oLAb IgG levels in the first and second trimester of pregnancy. Furthermore, we estimated the correlation between maternal serum (MS) levels of oLAb and alpha-1-fetoprotein (MS AFP), human chorionic gonadotrophin (MS HCG) and trophoblast-specific-beta-1-glycoprotein (MS SP1), because these proteins are determined as a part of prenatal biochemical screening for fetal congenital abnormalities. Our study deals with the oLAb changes in women with pregnancy-induced hypertension. We also investigated the correlation between oLAb IgG and anticardiolipin antibodies IgG (ACA) in the serum of pregnant women. We examined 40 pregnant women attending Institute for Mother and Child Care for their antenatal care as outpatients. Routine blood samplings between the 9-13th week of pregnancy and 16-18th week of pregnancy were performed as a part of biochemical prenatal screening for fetal congenital abnormalities (Group 1). Their mean age was 27 +/- 4.1 years. Furthermore, we examined 26 women in the second or third trimester with pregnancy-induced hypertension (Group 2). Group 2 was compared with 49 pregnant women in the second or third trimester who were normotensive (Group 3). We used commercial standardized ELISA kits for determination of oLAb IgG, ACA IgG, MS AFP and MS HCG, MS SP1 was analyzed by single radial immunodiffusion. We did not find any differences in the levels of oLAb IgG in the first and second trimester in the women of Group 1. The correlation between oLAb and ACA IgG was not statistically significant (Spearman coefficient r=0.22, p=0.1). The correlation between oLAb IgG with MS AFP, MS HCG and MS SP1 was not statistically significant. Weak negative correlation for AFP and HCG was suggested both in the first and in the second trimester. The levels of oLAb IgG in the group of women with pregnancy-induced hypertension were significantly lower than in the group of normotensive women (348 +/- 388 U/ml v.s. 579 +/- 400 mU/ml, p<0.01). We can conclude that the levels of oLAb do not differ in the first and second trimester of gravidity. However, we cannot exclude the possible influence of an inverse relationship between oLAb IgG titers and the synthesis of fetoplacental antigens. This finding is important especially in the context of the results of prenatal biochemical screening. Pregnancy-induced hypertension is associated with lower levels of oLAb. Weak cross-reactivity between oLAb and anticardiolipin antibodies may exist but there is a possibility that there are two different populations of antibodies reacting with various antigens.

[Advanced oxidation protein products in pregnancy]. / Produkty pokrocilé oxidace proteinu v tehotenství.

OBJECTIVE: Pregnancy and mainly its complications are associated with increased oxidative stress. Advanced oxidation protein products (AOPP) can serve as one of its markers. SETTING: First Institute of Medical Chemistry and Biochemistry and Institute for Clinical Biochemistry, First Medical Faculty, Charles University; Institute for Care of Mother and Child, Prague. METHODS: Together with parameters of prenatal screening, AOPP were measured in the serum of 23 pregnant women in the 2nd trimester of pregnancy. A group of healthy blood donors--women and men was used for comparison. AOPP were determined spectrophotometrically according to Witko-Sarsat (absorbance at 340 nm) and are expressed in chloramin units (mumol/l). RESULTS: Serum AOPP concentrations in pregnant women are significantly higher in comparison with blood donors--women (85.90 +/- 18.70 mumol/l vs 57.34 +/- 16.31 mumol/l, P < 0.0001) but there is no statistically significant difference between pregnant women and blood donors--men (85.90 +/- 18.70 mumol/l vs 78.60 +/- 44.01 mumol/l). AOPP level does not correlate either with the age of pregnant women or with the parameters of prenatal screening (human chorionic gonadotrophin--HCG, alpha-1-fetoprotein--AFP and trophoblast-specific--beta-1-glycoproteion--SP1). CONCLUSION: AOPP as a marker of oxidative stress is increased in the serum of pregnant women in comparison with women--blood donors but is similar as in men--blood donors which supports the hypothesis of hormonal influence. Nevertheless, AOPP do not correlate with the parameters of prenatal screening (HCG, AFP and SP1).

[PAPP-A in the first trimester of pregnancy]. / PAPP-A v prvním trimestru tehotenství.

OBJECTIVE: Pregnancy-associated plasma protein A has been reported to be low in Down syndrome affected pregnancies during the first trimester of pregnancy. The aim of this study was to determine preliminary the medians of pregnancy-associated plasma protein A (PAPP-A) in the first trimester of pregnancy and to compare PAPP-A with other biochemical markers used for biochemical prenatal screening. DESIGN: Retrospective study. SETTING: First Institute of Medical Chemistry and Biochemistry and Institute for Clinical Biochemistry, First Medical Faculty, Charles University. Institute for Care of Mother and Child, Prague. PATIENTS: One hundred forty one pregnant women, who undergo biochemical prenatal screening for chromosomal disorders between 7th and 13th week were studied. In addition six women in the second trimester and five women with twin pregnancies, two cases of trisomy 21 and one case of trisomy 18 in second trimester were available for study. METHODS: Maternal serum levels of PAPP-A, human chorionic gonadotropin (hCG) and alfa-1-feto-protein (AFP) were measured using ELISA methods. A single radial immunodiffusion was used to determine trophoblast-specific-beta-1-glycoprotein (SP1). RESULTS: PAPP-A levels increased throughout the first trimester with median 1.8 mg/l in the 7th week to 23.0 mg/l in the 13th week of pregnancy. PAPP-A serum levels from 3 women with twin pregnancies were higher than in women with singleton pregnancies. Serum levels of PAPP A in two women with fetus affected by chromosomal disorders did not differ from normal pregnancies. Correlation coefficients between PAPP-A and AFP and between PAPP-A and SP1 were statistically significant (r = 0.42, P < 0.001, respectively r = 0.54, P < 0.001). The levels of PAPP-A and HCG did not correlate significantly (r = 0.019, P = 0.82). CONCLUSION: We established first trimester medians for PAPP-A, which are necessary for evaluation of the pathological values. We found statistically significant correlation between PAPP-A and SP1 and PAPP-A and AFP.

Ethanol and oxidative stress.

This article represents the proceedings of a workshop at the 2000 ISBRA Meeting in Yokohama, Japan. The chair was Albert Y. Sun. The presentations were (1) Ethanol-inducible cytochrome P-4502E1 in alcoholic liver disease, by Magnus Ingelman-Sundberg and Etienne Neve; (2) Regulation of NF-kappaB by ethanol, by H. Matsumoto, Y. Nishitani, Y. Minowa, and Y. Fukui; (3) Chronic ethanol consumption increases concentration of oxidized proteins in rat liver, by Shannon M. Bailey, Vinood B. Patel, and Carol C. Cunningham; (4) Antiphospholipids antibodies and oxidized modified low-density lipoprotein in chronic alcoholic patients, by Tomas Zima, Lenka Fialova, Ludmila Mikulikova, Ptr Popov, Ivan Malbohan, Marta Janebova, and Karel Nespor; and (5) Amelioration of ethanol-induced damage by polyphenols, by Albert Y. Sun and Grace Y. Sun.

Oxidative stress, metabolism of ethanol and alcohol-related diseases.

Alcohol-induced oxidative stress is linked to the metabolism of ethanol. Three metabolic pathways of ethanol have been described in the human body so far. They involve the following enzymes: alcohol dehydrogenase, microsomal ethanol oxidation system (MEOS) and catalase. Each of these pathways could produce free radicals which affect the antioxidant system. Ethanol per se, hyperlactacidemia and elevated NADH increase xanthine oxidase activity, which results in the production of superoxide. Lipid peroxidation and superoxide production correlate with the amount of cytochrome P450 2E1. MEOS aggravates the oxidative stress directly as well as indirectly by impairing the defense systems. Hydroxyethyl radicals are probably involved in the alkylation of hepatic proteins. Nitric oxide (NO) is one of the key factors contributing to the vessel wall homeostasis, an important mediator of the vascular tone and neuronal transduction, and has cytotoxic effects. Stable metabolites--nitrites and nitrates--were increased in alcoholics (34.3 +/- 2.6 vs. 22.7 +/- 1.2 micromol/l, p < 0.001). High NO concentration could be discussed for its excitotoxicity and may be linked to cytotoxicity in neurons, glia and myelin. Formation of NO has been linked to an increased preference for and tolerance to alcohol in recent studies. Increased NO biosynthesis also via inducible NO synthase (NOS, chronic stimulation) may contribute to platelet and endothelial dysfunctions. Comparison of chronically ethanol-fed rats and controls demonstrates that exposure to ethanol causes a decrease in NADPH diaphorase activity (neuronal NOS) in neurons and fibers of the cerebellar cortex and superior colliculus (stratum griseum superficiale and intermedium) in rats. These changes in the highly organized structure contribute to the motor disturbances, which are associated with alcohol abuse. Antiphospholipid antibodies (APA) in alcoholic patients seem to reflect membrane lesions, impairment of immunological reactivity, liver disease progression, and they correlate significantly with the disease severity. The low-density lipoprotein (LDL) oxidation is supposed to be one of the most important pathogenic mechanisms of atherogenesis, and antibodies against oxidized LDL (oxLDL) are some kind of epiphenomenon of this process. We studied IgG oxLDL and four APA (anticardiolipin, antiphosphatidylserine, antiphosphatidylethanolamine and antiphosphatidylcholine antibodies). The IgG oxLDL (406.4 +/- 52.5 vs. 499.9 +/- 52.5 mU/ml) was not affected in alcoholic patients, but oxLDL was higher (71.6 +/- 4.1 vs. 44.2 +/- 2.7 micromol/l, p < 0.001). The prevalence of studied APA in alcoholics with mildly affected liver function was higher than in controls, but not significantly. On the contrary, changes of autoantibodies to IgG oxLDL revealed a wide range of IgG oxLDL titers in a healthy population. These parameters do not appear to be very promising for the evaluation of the risk of atherosclerosis. Free radicals increase the oxidative modification of LDL. This is one of the most important mechanisms, which increases cardiovascular risk in chronic alcoholic patients. Important enzymatic antioxidant systems - superoxide dismutase and glutathione peroxidase - are decreased in alcoholics. We did not find any changes of serum retinol and tocopherol concentrations in alcoholics, and blood and plasma selenium and copper levels were unchanged as well. Only the zinc concentration was decreased in plasma. It could be related to the impairment of the immune system in alcoholics. Measurement of these parameters in blood compartments does not seem to indicate a possible organ, e.g. liver deficiency.

Prevalence of various antiphospholipid antibodies in pregnant women.

Antiphospholipid antibodies (APAs) are characterized as a heterogeneous population of autoantibodies directed against different target antigens, predominantly anionic phospholipids or phospholipid-containing structures. The presence of APAs has been strongly associated with a variety of clinical disorders including adverse pregnancy complications such as spontaneous abortions, pregnancy-induced hypertension, preeclampsia and intrauterine growth retardation. The purpose of this study was to compare the prevalence of anticardiolipin antibodies (ACAs), which are routinely examined, with APAs directed against phosphatidylserine (APS), phosphatidylinositol (API), phosphatidylethanolamine (APE) and phosphatidylcholine (APC) in the sera of pregnant women. We examined 410 serum samples of pregnant women hospitalized in the department for pathological pregnancies. They underwent prenatal biochemical screening of fetal congenital abnormalities in the first and the second trimester of gravidity. Anticardiolipin IgG and IgM were measured using commercial ELISA kits (ImmuLisa Anti-Cardiolipin Antibody), whereas APS, APE, API and APC were determined by our modified ELISA kit. Among 410 pregnant women we found 21 patients (5.1%) positive for ACA IgG (>20 GPL) and 30 patients (7.3%) positive for ACA IgM (>10 MPL). It was found that 7.8% of pregnant women had at least one high-titer APA IgG and 9.8% high-titer APA IgM. One third of ACA IgG or IgM positive sera contained polyspecific autoantibodies reactive to at least two various phospholipids. In the group of IgG ACA positive women, 28.6% patients were positive for APS, 28.6% were positive or moderately positive for API, 23.8% for APC and 19% for APE. In the group of IgM ACA positive women, 33.3% were also positive for APS, 26.7% for APE, 26.7% for API and 23.3% for APC were present. IgG and IgM ACA negative patients exhibited a significantly lower incidence of other APA than the group of ACA positive pregnant women. It still remains to clarify if the routine examination of APA reacting with other anionic and zwitterionic antigens other than cardiolipin would improve the probability of identifying women liable to adverse pregnancy complications.

[Comparative study of the occurrence of beta-2-glycoprotein I antibodies (anti-beta-2-GPI) and anticardiolipin antibodies (ACA) in the blood of pregnant women]. / Srovnávací studie výskytu protilátek proti beta-2-glykoproteinu I (anti-beta-2-GPI) a antikardiolipinových protilátek (ACA) v séru tehotných zen.

OBJECTIVE: It is accepted now that most antiphospholipid antibodies (APA) are not directed against phospholipids alone but complexes of phospholipid-binding proteins and phospholipids. One of the phospholipid-binding proteins is beta 2-glycoprotein I (beta 2-GPI), a normal plasma glycoprotein, which under physiological conditions binds with negative charged phospholipids. Measurement of anti-beta 2-GPI antibodies requires specific tests, since ELISA for determination of anticardiolipin antibodies (ACA) may detect both antibodies directly binding cardiolipin and antibodies against cardiolipin-binding proteins. In this study a comparison of APAs against cardiolipin (CL), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylethanolamine (PE) v.s. anti-beta 2-GPI were compared. SETTING: First Institute of Medical Chemistry and Biochemistry, First Medical Faculty, Charles University and Institute for Care of Mother and Child. DESIGN: Retrospective clinical study using stored sera for determination of APA. METHODS: One hundred twenty four women in the first and the second trimester, who undergo biochemical prenatal screening for chromosomal disorders by maternal serum alpha-fetoprotein (MS AFP), human chorionic gonadotrophin (MS HCG) and trophoblast-specific beta 1-glycoprotein (MS SP1) were studied. In serum samples antibodies against CL, PS, PE, PI (isotype IgG and IgM) were examined by solid ELISA. 19 women who were positive at least for one type of APAs were selected and in the serum samples the levels of anti-beta 2-GPI were measured. RESULTS: No pregnant woman with anti-beta 2-GPI had positive antibodies against other phospholipids except CL. There was no significant correlation between levels of ACA IgG and IgM v.s. anti-beta 2-GPI IgG and IgM. Eight percent of serum samples were positive for both anti-beta 2-GPI IgG and ACA IgG. In the second trimester a statistically significant decrease of ACA IgG was found (p = 0.014). The difference of ACA IgM and anti-beta 2-GPI IgG and IgM was not statistically significant. Levels of foetoplacental antigens in anti-beta 2-GPI positive pregnant women were normal. CONCLUSION: A correlation between anti-beta 2-GPI antibodies and ACA was not found in our study. Anti-beta 2-GPI antibodies and ACA may be two different subpopulations of APA.

Antibodies against phospholipids and oxidized LDL in alcoholic patients.

Antiphospholipid antibodies (APA) are a generic term describing antibodies that recognize various phospholipids. Hepatocyte damage is a cardinal event in the course of alcoholic liver injury and autoantibodies against phospholipids could play an important role in this process. APA in alcoholic patients seem to reflect membrane lesions, impairment of immunological reactivity, liver disease progression and they correlate significantly with disease severity. LDL oxidation is supposed to be one of the most important pathogenic mechanisms of atherosclerosis and antibodies against oxidized low-density lipoprotein (oxLDL) are some kind of an epiphenomenon of this process. The scope of our study was to determine some autoantibodies (IgG-oxLDL and antiphospholipid antibodies) and their possible changes in alcoholic patients. We studied IgG-oxLDL and four APA - anticardiolipin antibodies (ACA), antiphosphatidylserine antibodies (APSA) antiphosphatidylethanolamine antibodies (APE) and antiphosphatidylcholine antibodies (APCA) in 35 alcoholic patients with mildly affected liver function at the beginning of the abuse treatment. The control group consisted of 60 healthy blood donors. In the studied group, we obtained positive results concerning total ACA in 17.1 % of alcoholic patients (8.3 % in the control group), 11.4 % IgG-ACA (6.7 %), 8.6 % IgM-ACA (3.3 %), 14.3 % total APE (6.7 %), 14.3 % total APCA (8.3 %) and 20 % total APSA (8.3 % in the control group). The IgG-oxLDL (406.4+/-52.5 vs 499.9+/-52.5 mU/ml) was not affected in alcoholic patients. We conclude that the autoantibodies against oxLDL are present in sera of alcoholics and healthy blood donors. Based on our results which revealed a wide range of IgG-oxLDL titres in the healthy population, this parameter does not appear to be very promising for the evaluation of the risk of atherosclerosis. Alcoholics with only mild affection of liver functions did not exhibit a significantly higher prevalence of all studied antiphospholipid antibodies (ACA, APSA, APE, APCA) which could lead to membrane lesions in these patients.

[Anti-phosphatidylserine antibodies in women with reproductive disorders]. / Antifosfatidylserinové protilátky u zen s poruchami reprodukce.

Antiphosphatidylserine antibodies (APSA) belong to the heterogeneous population of antiphospholipid antibodies (APA) which are oriented above all against negatively charged phospholipids. The presence of APA in women is closely associated with repeated miscarriages and other complications during pregnancy. The most frequently detected specific antibodies in these patients are autoantibodies against cardiolipin and phosphatidylserine (PS). In a group of 84 pregnant women where within the framework of biochemical prenatal screening of inborn developmental defects serum levels of alpha-1-fetoprotein, choriogonadotropin and trophoblast specific beta-1-glycoprotein were examined as well as in 22 women treated for primary sterility and 22 blood donors the authors assessed, using the ELISA method, antiphosphatidylserine and cardiolipin antibodies (ACA). They found an increased prevalence of APSA in all examined groups as compared with the control group of blood donors. In pregnant women the prevalence of APSA and ACA did not differ and at least one type of antibodies was detected in 20.1%. In pregnant women with positive APSA in the case-records spontaneous abortions were recorded, or imminent abortions during the present gestation or treatment on account of sterility, and in some instances also changes of foetoplacental antigen serum levels were found. It is therefore likely that the presence of APA in women may be one of the factors participating in reproductive disorders and that assessment of APSA together with APA may extend the spectrum of immunological examinations, in particular in sterile and infertile women.

[ELISA in the determination of antiphosphatidylserine antibodies]. / Metoda ELISA pro stanovení antifosfatidylserinových protilátek.

Antiphospholipid antibodies (APA) are a group of antibodies against various phospholipid antigens. In order to extend the spectrum of examined specificities of antiphospholipid antibodies the authors elaborated an ELISA method for assessment of antiphosphatidyl serine antibodies (APSA). As antigen they used phosphatidyl serine isolated from the white matter of cattle brain. The ELISA method was tested by examining APSA in 12 patients with rheumatic diseases, 24 women with reproductive disorders and 50 patients with testicular tumours and the results were compared with examinations of anticardiolipin antibodies. The concurrent presence of both types of antibodies was recorded in 20.8% women with reproductive disorders and in 14% of the patients with testicular tumours. In these groups antiphosphatidyl serine antibodies were found more frequently.

[Anti-phosphatidylinositol antibodies and their determination using our ELISA method]. / Antifosfatidylinositolové protilátky a jejich stanovení vlastní ELISA metodou.

Antiphospholipid antibodies (APA) are autoantibodies to negatively charged phospholipids such as a cardiolipin, phosphatidylserine and phosphatidylinositol. The occurrence of APA is associated with arterial and venous thromboembolic manifestations, thrombocytopenia and recurrent fetal loss. To date most studies have concentrated on antibodies to cardiolipin specifically. In this study we present own modification of the ELISA method for assessment of antiphosphatidylinositol antibodies (API). The phosphatidylinositol isolated from small green peas was used as an antigen. To test ELISA method 151 serum samples of patients with various diseases and 22 serum samples of blood donors were examined and the results were compared with the results of anticardiolipin antibodies (ACA). The positivity of API was 22.5% and ACA 20.5% in a group of patients. The simultaneous of both types of APA was found in 15.2%.

[Antiphospholipid antibodies in patients with neurologic diseases]. / Antifosfolipidové protilátky u pacientu s vybranými neurologickými chorobami.

In a series of patients with neuroinfection, Lyme disease, Guillain Barré syndrome, demyelinization, partial or generalized, epilepsy, we have investigated antiphospholipid antibodies of IgG and IgM subtypes, together with anticoagulant factors, member of thrombocytes, sedimentation rate of erythrocytes. Coagulant factor disorder in primary acute inflammatory processes (attack of demyelinization, neuroinfection, acute viral respiratory infection) and primary thrombocyte disorder (focal epilepsies) may be a result of cross reaction of antiphospholipid antibodies with negative polarized phospholipids in membranes of central nervous system, endothelium of cerebral and extracranial vessels and in the inner thrombocyte membranes connected with prostaglandin production disorder.

[Anticardiolipin antibodies and oncofetal antigens in pregnant women]. / Antikardiolipinové protilátky a onkofetální antigeny u tehotných zen.

The presence of antiphospholipid antibodies (APA) in the blood stream is associated with some reproductive disorders in women. In the serum of 84 pregnant women, using the Elisa method, anticardiolipin antibodies (ACA) were examined. The first group of 53 women was formed by women with pathological levels of at least one of the oncofoetal antigens (alpha-1-fetoprotein-AFP, human chorionic gonadotropin-HCG and trophoblast-specific beta-1-glycoprotein- SP1). The second group of 31 pregnant women comprised women hospitalized on account of a risk pregnancy. In the first group women with reduced HCG were most frequently represented, where in 8.8% positive ACA were recorded. Of 11 patients with elevated HCG only in one the presence of ACA was detected. In nine pregnant women with reduced HCG and SP1 or AFP ACA positivity was found in 44.4%. In the second group ACA were detected in the serum of six pregnant women who had also pathological SP1 or HCG levels. From the results ensues that ACA could interfere with transduction signalizing processes in the cell and thus influence the synthesis of some proteins.

[Antiphospholipid antibodies in women with reproductive disorders]. / Antifosfolipidové protilátky u zen s nekterými poruchami reprodukce.

The authors examined in serum of 121 women, using the ELISA method, anticardiolipine antibodies (ACA) class IgG and IgM. The examined women were divided into a group of women with a normal pregnancy (n = 14), a group of women with imminent abortion during the first trimester (n = 10) and a group of women treated for prolonged periods on account of sterility (n = 97). In women with a normal pregnancy only in one a positive titre in the IgG class was recorded. In the group with imminent abortion a positive ACA titre was found in three patients (30%); in women with negative results of the ACA examination, pregnancy continued. In sterile women positive ACA tests were recorded in 14.5%.

[Determination of antiphospholipid antibodies in serum using ELISA]. / Stanovení antifosfolipidových protilátek v séru metodami ELISA.

The authors compare two ELISA methods for the assessment of antiphospholipid antibodies, classes IgG and IgM, in serum: ELISA Pin Plate System ALPHA DIALAB Co. and the ELISA method developed in the Research Institute of Rheumatic Diseases. Both methods use cardiolipin as antigen. In the Pin Plate test the immunochemical reaction antigen/antibody does not take place at the surface of the pits of the microtitration plates but on the tip of the next plate. The results of examinations of antiphospholipid antibodies obtained by the tested methods are comparable, the Pin Plate test is quicker and more sensitive, but its price limits routine use.

Prenatal biochemical diagnostics of inborn developmental defects.

Screening of pathological pregnancies with the use of biochemical markers determined in maternal serum is now widely accepted as a useful procedure. In our experience, the main contribution is a finding of abnormal values of one or more of the markers, which will advise gynecologist upon a possibility of a risk pregnancy.

[SP1 serum levels in pregnancy with Rh isoimmunization]. / Sérové hladiny SP1 u tehotenství s Rh izoimunizací.

The authors examined in 37 pregnant women hospitalized on account of Rh incompatibility during the second and third trimester 71 serum specimens for levels of trophoblast specific beta 1-glycoprotein (SP1), using the method of simple radial immunodiffusion. In the group of 16 women with Rh incompatibility whose pregnancy terminated by delivery of a normal neonate, in 22% the SP1 levels were higher than 1.5 x the median value for the given gestation week (MoM). In 17 pregnancies where the foetus suffered from haemolytic disease the SP1 levels were higher than 1.5 MoM in 45% of the examined foetuses. Markedly elevated SP1 levels of 2.3 and 2.4 MoM were recorded in two pregnancies with hydrops of the foetus.