Precision of the retinal tomograph as a screening device.

PURPOSE: To assess the diagnostic accuracy of the Heidelberg Retinal Tomograph 3 (HRT3) as a screening device in comparison with the reference standard of Octopus standard automated perimetry results (SAP) combined with clinical findings. METHODS: All patients underwent screening examinations and investigations within a single day. Abnormal screening results were classified as follows: The HRT3: Either "borderline" or "outside normal limits" using the global Moorfields classification (MFC); SAP and clinical exam: A mean defect>2.4 dB or "outside normal limits" clear text analysis of SAP; and one of the following i) IOP>21 mmHg, ii) Van Herrick<», iii) cup disc ratio>0.55, iv) optic nerve head abnormality, v) narrow iridocorneal angle or vi) evidence of peripheral anterior synechiae on gonioscopy. RESULTS: The mean age of the participants was 59.9 years (±14.8 [21, 91]). Twenty-three subjects (16%) were classified as abnormal on SAP and clinical exam. The HRT3 classification had a sensitivity of 30% (95% CI [16%, 51%]) with associated specificity of 58% (95% CI [49%, 66%]). Of the sixty subjects classified as borderline or outside normal limits with the HRT MFC global result, seven subjects were also abnormal according to SAP and clinical exam. CONCLUSION: The results suggest that the HRT3 may not be suitable as a sole screening device; however, further investigation is necessary.

Biological effects of conjugated linoleic acids supplementation.

Conjugated linoleic acids (CLA) are a group of polyunsaturated fatty acids (PUFA) with a single pair of conjugated double bonds. The major natural CLA isomer is 18:2 cis-9, trans-11 (c9, t11) linoleic acid, or rumenic acid (RA). Chemically synthesized CLA is also available, mostly as a mixture of RA and 18:2 trans-10, cis-12 (t10, c12) isomers in equal amounts (50:50). Consumption of ruminant meat (beef and lamb) and dairy products (milk and cheese) is the main source of dietary exposure to CLA. Despite numerous studies on animal and human models (tumorigenesis, obesity, immune response) it has not been established whether additional supplementation of CLA is of benefit. Moreover, some studies, conducted both in animals and in humans, reveal that CLA isomers may induce insulin resistance. Presently, balanced diet rich in CLA from natural sources is recommended. The purpose of this review was to sum up the results available in the literature.

OS007. The apoptosis markers are altered in CD4(+)CD25(+)FOXP3(+) T regulatorylymphocytes in pre-eclampsia.

INTRODUCTION: Pre-eclampsia (PE) is a common obstetric syndrome affecting about 5-10% of pregnant women. The etiology and pathogenesis of this syndrome are not fully understood. There are many studies describing alterations in the innate and adaptive immune system which may have an influence on the onset of this disorder. It was suggested that the activation of cell-mediated immunity may play the key role in the etiology of pre-eclampsia. It was proposed that inappropriate activation of the immune system can lead to pre-eclampsia. OBJECTIVES: The aim of our study was to estimate the surface expressions of CD95(APO-1/Fas) antigen and the intracellular expressions of anti-apoptotic proteinBcl-2 and pro-apoptotic proteinBax in CD4(+)CD25(+)FoxP3(+) T regulatory lymphocytes (Tregs) as well as the percentage of CD8(+)CD28(+) T cytotoxic cells in peripheral blood of patients with pre-eclampsia in comparison with healthy pregnant women in the third trimester of physiological pregnancy. METHODS: Twenty-four women with pre-eclampsia and twenty normal third trimester pregnant women were included in the study. The lymphocytes were isolated from peripheral blood samples and labelled with monoclonal antibodies. The expressions of surface antigens and intracellular proteins were estimated using flow cytometry. RESULTS: The population of CD4(+)CD25(+)FoxP3(+) Treg cells was significantly lower in peripheral blood of patients with pre-eclampsia when compared to normal third trimester pregnant women. The percentages of CD4(+)CD25(+)FoxP3(+) Treg cells that express Bcl-2 protein were significantly lower in peripheral blood of patients with pre-eclampsia when compared to healthy pregnant women, whereas the percentages of CD4(+)CD25(+)FoxP3(+) Treg cells with the expressions of Bax protein did not differ in both groups. Moreover, the mean fluorescence intensity (MFI) of Bcl-2 protein in CD4(+)CD25(+)FoxP3(+) Treg cells was significantly lower and MFI of Bax protein significantly higher in pre-eclampsia when compared to the control group. The percentage of CD8(+)CD28(+) T cells did not differ in both studied groups but MFI of CD28 antigen on T CD8(+) cells was significantly higher in pre-eclampsia when compared to the control group. CONCLUSION: The obtained results suggest that the deficit of CD4(+)CD25(+)FoxP3(+) Treg lymphocytes which is observed in pre-eclampsia maybe associated with alterations in apoptosis markers.

PP069. The expressions of B7-H1 and B7-H4 co-stimulatory molecules on myeloid and lymphoid dendritic cells in pre-eclampsia and normal pregnancy. The expressions of B7-H1 and B7-H4 co-stimulatory moleculeson myeloid and lymphoid dendritic cells in pre-eclampsia and normal pregnancy.

INTRODUCTION: Pre-eclampsia (PE) is a common obstetric syndrome affecting about 5-10% of pregnant women. The syndrome is a leading cause of maternal and fetal morbidity and mortality, even in developed world. The etiology and pathogenesis of this syndrome are not fully understood. There are many studies describing alterations in the innate and adaptive immune system which may have an influence on the onset of this disorder. It was suggested that activation of cell-mediated immunity may play the key role in the etiology of pre-eclampsia. OBJECTIVES: The purpose of our study was to estimate the expressions of B7-H1 and B7-H4 costimulatory molecules on myeloid and lymphoid DCs (CD1c(+), BDCA-2(+)) in the peripheral blood of patients with pre-eclampsia and normal pregnant women in the third trimesters of physiological pregnancy. METHODS: Thirty three patients with pre-eclampsia and 26 normal pregnant women were included in the study. Dendritic cells were isolated from peripheral blood, stained with monoclonal antibodies against blood dendritic cell antigens and B7-H1 and B7-H4 molecules and estimated using flow cytometry. RESULTS: The expressions of B7-H1 molecule on CD1c(+) myeloid DCs and B7-H4 molecule on BDCA-2(+) lymphoid DCs did not differ in pre-eclampsia and healthy third trimester pregnant women. The expressions of B7-H4 molecule on CD1c(+) myeloid DCs and B7-H1 molecule on BDCA-2(+) lymphoid DCs were significantly higher in peripheral blood of patients with pre-eclampsia in comparison with the control group. CONCLUSION: It seems possible that higher expressions of B7-H4 molecule on CD1c(+) myeloid DCs and B7-H1 molecule on lymphoid BDCA-2(+) DCs in pre-eclampsia may be the tolerogenic mechanism secondary to the pro-inflammatory response which is observed in this syndrome.

PP135. Maternal serum levels of endothelial nitric oxide synthase and ADMA, an endogenous ENOS inhibitor in pregnancies complicated by severe preeclampsia.

INTRODUCTION: Preeclampsia is a major cause of maternal and fetal mortality and morbidity. Preeclampsia remains amongst the biggest challenges in obstetrics, but its aetiopathogenesis is still unclear. Recent observations support the hypothesis that imbalance between maternal angiogenic/anti-angiogenic factors concentrations is responsible for the clinical manifestations of the preeclampsia. It was also suggested that clinical manifestations of preeclampsia caused by endothelial malfunction including insufficient production of nitric oxide and that abnormality in nitric oxide synthesis or NO bioavailability may contribute to the development of preeclampsia. But the mechanism associated with alteration of nitric oxide formation in pregnancies complicated by preeclampsia is not well understood. OBJECTIVES: The aim of this study was to determine the maternal serum concentrations of endothelial nitric oxide synthase (eNOS) and its endogenous inhibitor, asymmetric dimethylarginine (ADMA) in pregnancies complicated by severe preeclampsia in comparison with healthy normotensive pregnant women. METHODS: A study was conducted including 29 uncomplicated women (the Control group) and 29 preeclamptic women (the Pre group). Preeclampsia was diagnosed by the increased blood pressure of >140mmHg systolic and >90mmHg diastolic in women who were normotensive before 20 weeks of gestation accompanied by proteinuria, defined as the urinary excretion of >0.3g protein in 24h specimen. Severe preeclampsia was defined as blood pressure >160/110mmHg on at least 2 occasions 6h apart with proteinuria >5g in a 24h urinary protein excretion. The maternal serum eNOS and ADMA concentrations were determined using a sandwich enzyme-linked immunosorbent assays. RESULTS: There were no statistically significant differences in gravidity, parity, maternal age and height in patient profiles between groups. Creatinine and urea levels were normal in all patients. Systolic and diastolic blood pressures were higher in preeclamptic pregnant women than in the control group. The mean systolic blood pressure values were 166.39±15.47mmHg in the Pre group and 115.72±10.34mmHg in the control group. The mean diastolic blood pressure values were 108.89±10.45mmHg in the Pre group and 75.34±6.22mmHg in the healthy controls. These differences were statistically significant (p<0.000001). Our study revealed increased maternal serum levels of asymmetric dimethylarginine in preeclamptic patients (p=0.007223). The mean values of maternal serum ADMA were 0.597±0.161mmol/L in the Pre group versus0.503±0.081mmol/L in the healthy pregnant patients. The levels of serum endothelial nitric oxide synthase were lower in women with pregnancies complicated by severe preeclampsia than in the healthy women from the control group, but these differences were not statistically significant (p=0.118770). The mean values of serum eNOS were 134.06±76.73U/ml in the Pre group and 187.70±165.41U/ml in the Control group. CONCLUSION: Elevated levels of ADMA and the unchanged levels of eNOS in pregnancies complicated by severe preeclampsia allow the conclusion that the nitric oxide deficiency in this pregnancy disorder result not from a reduced level or activity of eNOS, but from elevated levels of asymmetric dimethylarginine, an endogenous eNOS inhibitor.

The expression of B7-H1 and B7-H4 molecules on immature myeloid and lymphoid dendritic cells in cord blood of healthy neonates.

The aim of our study was to estimate both B7-H1 and B7-H4 molecules on immature myeloid and lymphoid dendritic cells in umbilical cord blood of healthy neonates in comparison with peripheral blood of healthy adults. Thirty nine healthy full-term neonates from physiological single pregnancies and 27 healthy adults were included in the study. The expression of B7-H1 and B7-H4 was revealed using the immunofluorescence method. Statistical analysis was performed using a non-parametric test (Mann-Whitney U-Test). The percentages of BDCA-1+ dendritic cells with B7-H1 and B7-H4 expressions were significantly higher in peripheral blood of healthy adults (p<0.00003). It was either observed that the percentage of BDCA-2+ dendritic cells with the expression of B7-H4 molecules was significantly higher in peripheral blood of healthy adults in comparison with umbilical cord blood (p<0.02). Decreased percentages of dendritic cells and co-stimulatory molecules indicate that neonates have immature immune system. Depletion of co-stimulatory B7-H1 and B7-H4 molecules enable appropriate development of immune response.

The expression and concentration of CD40 ligand in normal pregnancy and pre-eclampsia.

This study investigated the surface expression of the CD154 antigen (CD40L) on peripheral blood CD4+ T lymphocytes and the sera concentrations of soluble CD40L antigens (sCD40L) in non-pregnant women, normal pregnant women, and patients with pre-eclampsia. Twenty-five patients with pre-eclampsia, 18 healthy pregnant women, and 10 healthy non-pregnant women were included in the study. The expression of the CD154 antigen on CD4+ T lymphocytes was determined using flow cytometry. The serum concentration of sCD40L was measured using an ELISA. Statistical analysis was performed using the Mann-Whitney U and ANOVA tests. The expression of the CD154 antigen on CD4+ T lymphocytes and the serum concentration of soluble CD40L were significantly higher in pre-eclampsia than in normal pregnant women. In normal pregnancy the expression of the CD154 antigen on CD4+ T lymphocytes and the concentration of sCD40L were significantly lower than in non-pregnant women. We conclude that during normal pregnancy the levels of these inflammatory mediators are lower than in non-pregnant women. In pre-eclampsia the levels are higher than those in normal pregnancy, but stable compared with the non-pregnant state. These results suggest that in pre-eclampsia, there are disturbances in the mechanisms responsible for the decrease in innate immunity which occurs in normal pregnancy.

Activated T lymphocytes in pre-eclampsia.

PROBLEM: The aim of our study was to investigate the activation markets of T CD3(+), T helper CD4(+) and T cytotoxic CD8(+) cells, as well as, the populations of T naïve CD4(+) CD45RA(+), T memory CD4(+) CD45RO(+) and T regulatory lymphocytes in PE and healthy pregnant women. METHOD OF STUDY: Twenty-five patients with PE and thirty healthy third trimester pregnant women were included in the study. Peripheral blood mononuclear cells were isolated from peripheral blood, stained with monoclonal antibodies and estimated using the flow cytometric method. RESULTS: The percentages of CD4(+)CD25(+), CD4(+)CD25(dim), CD3(+)HLA-DR(+), CD4(+)HLA-DR(+) and CD8(+)HLA-DR(+) cells did not differ between study groups. The population of T regulatory CD4(+)CD25(bright) lymphocytes was significantly lower in the group of patients with PE when compared with the controls (P < 0.01). The percentages of CD3(+)CD25(+) (P < 0.05), CD8(+)CD25(+) (P < 0.05), CD4(+)45RO(+) (P < 0.01) lymphocytes were significantly higher, while CD4(+)CD45RA(+) (P < 0.01) cells--significantly lower in peripheral blood of patients with PE when compared with the control group. CONCLUSION: The increased levels of T CD4(+)45RO(+) and T CD8(+) CD25(+) cells can suggest the activation of CD4(+) and CD8(+) T lymphocytes in pre-eclampsia. It seems possible that the activation of T lymphocytes is associated with the deficiency of T regulatory cells in PE.

Apoptosis of HeLa cell lines incubated with retinol.

PURPOSE: The aim of the study was to evaluate a soluble form of APO-1/Fas antigen in supernatants from HeLa cell line culture after 24 and 72 h of incubation with selected retinoic acid concentrations. MATERIALS AND METHODS: HPV18-positive cell lines were cultivated with All-trans-retinoic acid (ATRA) at concentrations of 1 x 10(-6) and 1 x 10(-9) M. The cultures were incubated for 24 and 72 h. A control culture with 3 microl of DMSO was incubated under identical conditions. The concentrations of soluble APO-1/Fas antigen in cell culture supernatants were estimated using an ELISA method. RESULTS: The culture with 72-h incubation with retinoic acid proved to be toxic to cells and was excluded from the analysis. The results obtained showed a significantly lower concentrations of soluble APO-1/Fas antigen in supernatants from cell lines incubated with retinol for 24 h than in the controls. CONCLUSIONS: The higher concentrations of soluble APO-1/Fas antigen in supernatants from the HeLa cell line without retinol may constitute a protective mechanism of the cells infected with the virus before undergoing Fas/FasL-dependent apoptosis. Lower concentrations of sAPO-1/Fas antigen in the supernatant from HeLa cell culture incubated with retinol may suggest that mechanisms protecting infected cells against Fas/FasL-mediated apoptosis become defective under the influence of retinol. Our studies confirm that Vitamin A and its analogues inhibit the proliferation of cells associated with HPV infection and suggest promising effects of retinoid therapy in inhibiting the progression of early cervical lesions to cancer.

Evaluation of placental angiotensin type 1 receptors in women with hypertension during pregnancy.

OBJECTIVE: The aim of this study was to examine and compare the localization and immunoreactivity of the angiotensin type-1 receptor (AT1R) in human placental tissue in patients with pregnancy complicated by pre-eclampsia with and without intrauterine growth restriction (IUGR). METHODS: Immunohistochemistry was used to examine the localization of the AT1R in human placental tissue. Semiquantitative immunohistochemical H-score values for each placental cell type were calculated. RESULTS: Elevated H-score index values for decidual cells, syncytiotrophoblast and cytotrophoblast in pre-eclamptic patients were found to be higher in comparison with values in the control group. Decreased H-score index values for syncytiotrophoblast and cytotrophoblast were found in patients with pregnancy complicated by IUGR in the course of pre-eclampsia. CONCLUSIONS: The results obtained in this study point out the significant role of the AT1R and suggest that normal AT1R activity is of fundamental significance for the normal course of pregnancy and proper fetal growth.

CD1c(+) immature myeloid dendritic cells are predominant in cord blood of healthy neonates.

It has been suggested lately that some types of antigen presenting cells-myeloid dendritic (DC-1) cells can differentiate the immune response towards Th1 type immunity, whereas lymphoid cells (DC-2) can stimulate Th2 type immunity. It has been observed that neonates are deficient in Th1 response. The purpose of our study was to estimate the proportions of immature myeloid (CD1c(+)) and lymphoid (BDCA-2(+), BDCA-4(+)) dendritic cells and the CD1c(+):BDCA-2(+) cell ratio in cord blood of healthy neonates in comparison with dendritic cells of healthy adults. Thirty healthy neonates born from normal pregnancies and 30 healthy adults were included in the study. The dendritic cells were isolated from cord and peripheral blood, stained with anti-CD1c, anti-BDCA-2, anti-BDCA-4, anti-CD123 and anti-CD19 monoclonal antibodies and estimated using flow cytometry. The percentage of CD1c(+) dendritic cells in cord blood of healthy newborns did not differ significantly when compared to those in peripheral blood of healthy adults. The percentages of cord blood BDCA-2(+) and BDCA-4(+) dendritic cells of neonates were significantly lower when compared to lymphoid dendritic cells in peripheral blood of adults. The CD1c(+):BDCA-2(+) ratio was significantly higher in cord blood of neonates in comparison with CD1c(+):BDCA-2(+) ratio in adult's blood. Myeloid and lymphoid dendritic cells may be involved in the immune regulation during fetal development. Immature myeloid dendritic cells are predominant in cord blood of healthy neonates. Immature lymphoid dendritic cells are not the major population of dendritic cells in cord blood.

Blood myeloid and lymphoid dendritic cells are stable during the menstrual cycle but deficient during mid-gestation.

The aim of this study was to estimate the populations of peripheral blood myeloid and lymphoid dendritic cells (CD1c(+), BDCA-2(+), BDCA-4(+)) and the CD1c(+):BDCA-2(+) ratio in phases of the ovarian cycle and in normal pregnant patients. 18 non-pregnant women and 17 normal pregnant women were included. Dendritic cells were isolated from peripheral blood, stained with monoclonal antibodies (mAbs) against blood dendritic cell antigens (anti-BDCA-1, BDCA-2, BDCA-4) and estimated using flow cytometry. CD1c(+), BDCA-2(+) and BDCA-4(+) dendritic cells were present in the follicular and luteal phases of the ovarian cycle and in all trimesters of normal pregnancy. The percentages of CD1c(+) dendritic cells did not differ between the follicular and luteal phases of the ovarian cycle. The percentage of BDCA-2(+) dendritic cells was lower in the luteal phase of the ovarian cycle compared with the follicular phase, but the differences were not statistically significant. The CD1c(+):BDCA-2(+) cell ratio was significantly lower in the luteal phase compared with the follicular phase of the ovarian cycle. The numbers of dendritic cells were significantly lower in the second trimester when compared with the first and third trimesters of normal pregnancy. Furthermore, in the second trimester, the CD1c(+):BDCA-2(+) ratio was higher than in the other trimesters of normal pregnancy. All populations of dendritic cells and the CD1c(+):BDCA-2(+) ratio did not differ in the first and third trimesters of physiological pregnancy. Our results suggest that myeloid and lymphoid dendritic cells are not affected by steroid hormones during the menstrual cycle. The deficiency of peripheral blood dendritic cells observed during the second trimester of normal pregnancy can be associated with their migration to the uterus during the second physiological invasion by cytotrophoblast.

Myeloid and lymphoid dendritic cells in normal pregnancy and pre-eclampsia.

The aim of our study was to estimate the populations of peripheral blood myeloid and lymphoid dendritic cells (CD1c+, BDCA-2+) and the CD1c+ : BDCA-2+ ratio in normal pregnant women and in patients with pre-eclampsia. Fifteen women in the first, second and third trimesters of normal pregnancy, and 25 patients with pre-eclampsia were included in the study. The dendritic cells were isolated from peripheral blood, stained with monoclonal antibodies against blood dendritic cell antigens (anti-CD1c, anti-BDCA-2) and estimated using the flow cytometric method. CD1c+ and BDCA-2+ dendritic cells were present in women during all trimesters of physiological pregnancy and in pre-eclamptic patients. It was observed that the numbers of dendritic cells were significantly lower in the second trimester when compared with the first and third trimesters of normal pregnancy. Furthermore, in the second trimester, CD1c+ : BDCA-2+ ratio was higher than in the other trimesters of physiological pregnancy. All populations of dendritic cells and CD1c+ : BDCA-2+ ratio did not differ in the first and third trimesters of normal pregnancy. The percentage of BDCA-2+ dendritic cells was significantly lower in pre-eclampsia in comparison with healthy women in the third trimester of physiological pregnancy, while CD1c+ : BDCA-2+ ratio was significantly higher in pre-eclamptic patients when compared with control groups. We concluded that dendritic cells may be involved in the immune regulation during physiological pregnancy. CD1c+ and BDCA-2+ cells can influence the Th2 phenomenon which is observed during physiological pregnancy. Furthermore, it seems possible that lower BDCA-2+ cells percentage and higher CD1c+ : BDCA-2+ ratio can be associated with increased Th1-type immunity in patients with pre-eclampsia.

Intrapartum cardiotocography and fetal pulse oximetry in assessing fetal hypoxia.

OBJECTIVES: A retrospective analysis of short-term variability (STV), a cardiotocography (CTG) parameter, in relation to fetal blood saturation values (FSpO(2)) obtained by fetal pulse oximetry. METHODS: The study included 26 healthy pregnant women monitored continuously during delivery with both cardiotocography and fetal pulse oximetry. RESULTS: Lower FSpO(2) values were observed in the group showing STV levels 6.0 ms (34.4+/-2.9% vs. 43+/-7.2%; P<0.001). A positive correlation was found between STV levels

Circadian rhythm chaos: a new breast cancer marker.

The most disappointing aspect of breast cancer treatment as a public health issue has been the failure of screening to improve mortality figures. Since treatment of late-stage cancer has indeed advanced, mortality can only be decreased by improving the rate of early diagnosis. From the mid-1950s to the mid-1970s, it was expected that thermography would hold the key to breast cancer detection, as surface temperature increases overlying malignant tumors had been demonstrated by thermographic imaging. Unfortunately, detection of the 1-3 degrees C thermal differences failed to bear out its promise in early identification of cancer. In the intervening two-and-a-half decades, three new factors have emerged: it is now apparent that breast cancer has a lengthy genesis; a long-established tumor-even one of a certain minimum size-induces increased arterial/capillary vascularity in its vicinity; and thermal variations that characterize tissue metabolism are circadian ("about 24 hours") in periodicity. This paper reviews the evidence for a connection between disturbances of circadian rhythms and breast cancer. Furthermore, a scheme is proposed in which circadian rhythm "chaos" is taken as a signal of high risk for breast cancer even in the absence of mammographic evidence of neoplasm or a palpable tumor. Recent studies along this line suggest that an abnormal thermal sign, in the light of our present knowledge of breast cancer, is ten times as important an indication as is family history data.