Fibronectin, plasminogen activator inhibitor type 1 (PAI-1) and uterine artery Doppler velocimetry as markers of preeclampsia

Objective: The purpose of this study was to examine plasma levels of fibronectin and plasminogen inhibitor type 1 (PAI-1), and alterations in uterine artery (UtA) waveforms throughout normotensive and preeclamptic pregnancies and to analyze its predictive value for the detection of preeclampsia within the second trimester of pregnancy. Material and methods: Blood samples were collected from 102 healthy, nulliparous women between the 24th and 26th gestational week. Preeclampsia developed in 13 patients; 89 normotensive control subjects were matched from the same cohort. Plasma samples were assayed for fibronectin and PAI-1 by enzyme-linked immunosorbent assay. Color pulsed Doppler examinations of UtA were performed after blood sampling. Trends were compared between two groups. Results: Maternal plasma fibronectin and PAI-1 levels and average PI, RI and S/D ratios of patients with preeclampsia were significantly higher (p < 0.05). The best cut-off values for predicting preeclampsia of fibronectin, PAI-1, PI, RI, S/D ratio based on ROC curve analysis were 290 mg/ml, 77.3 ng/ml, 1,0615, 0.605 and 2,59 respectively. The areas under the curve equal to 0.705, 0.753, 0.689, 0.695 and 0.699 for fibronectin, PAI-1 and uterine artery Doppler PI, RI, S/D ratio were determined for the prediction of preeclampsia. Conclusions: Fibronectin, PAI-1 and UtA Doppler are potentially useful predictors of preeclampsia. Maternal plasma PAI-1 combinated with fibronectin had the highest predictive values in our study.     

Soluble Intercellular Adhesion Molecule-1 Serum Profile in Physiologic and Preeclamptic Pregnancy

PROBLEM: The aim of this study was to determine serum levels of soluble intercellular adhesion molecule (sICAM)-1, an adhesion receptor that mediates interactions with the immune system, in physiologic and preeclamptic pregnancies. Moreover, we evaluated whether the release of sICAM-1 during pregnancy correlated to plasma fibronectin concentrations. METHOD OF STUDY: Serum was collected from 18 nonpregnant, control women, from 58 normal pregnant women during the first (n = 13), second (n = 15), and third (n = 30) trimesters, and from 25 preeclamptic patients at 27–39 weeks' gestation. All samples were assayed for sICAM-1 by a specific enzyme-linked immunoassay and for fibronectin by a nephelometric system. Serum sICAM-1 levels in preeclamptic patients were compared to those obtained from gestational-matched normal pregnant women. RESULTS: Levels of sICAM-1 were significantly elevated (P < 0.001) in each of the three trimesters of normal pregnancy (I trimester: 390.4 ± 25.7 ng/ml; II trimester: 386.3 ± 15.4 ng/ml; and III trimester: 367.3 ± 15.8 ng/ml) when compared to those of healthy nonpregnant women (263.3 ± 11.6 ng/ml). No significant difference in sICAM-1 concentrations was observed among the three trimesters. Preeclampsia was associated to a significant decrease (P < 0.01) of sICAM-1 levels (309.8 ± 11.6 ng/ml) relative to those observed in gestational-matched pregnant women (367.3 ± 15.8 ng/ml). Fibronectin and sICAM-1 levels did not correlate. CONCLUSION: The increased levels of sICAM-1 found in physiologic pregnancies and its reduction in preeclampsia may account for some of the immunologic alterations demonstrated to be associated with pregnancy.     

Plasma basic fibroblast growth factor levels in colorectal cancer: a clinically useful assay?

Angiogenic cytokines in the plasma and serum of cancer patients may serve as `surrogate' markers of tumour neoangiogenesis. Serum VEGF correlates with disease stage in colorectal cancer (CRC), but the role of bFGF in CRC is uncertain. This study aimed to assess plasma bFGF levels in CRC patients before treatment, during chemoradiotherapy and at one-year follow-up. Plasma samples were taken from 124 CRC patients, 26 polyp patients and 55 controls, and bFGF levels were measured by ELISA. 19 patients underwent pre-operative chemoradiotherapy. One-year follow-up samples were available from 48 disease-free patients and 18 patients with progressive disease. There were no detectable differences between plasma bFGF levels in polyp, Dukes' A or B patients (4.55, 5.77, 4.25 pg/ml, respectively), but there was a significant increase in metastatic CRC patients [Dukes' C and D (7.42 and 6.6 pg/ml; P=0.004 and 0.048, respectively)], relative to median control levels of 4.14 pg/ml. At follow-up, there was a significant fall in plasma bFGF levels in disease-free patients (pre-op 6.09 and follow-up 3.45 pg/ml, P=0.0004), but a non-significant rise in 18 patients with progressive disease (pre-treatment 5.90 and follow-up 9.99 pg/ml, P=0.33). Pre-treatment plasma bFGF in patients receiving chemo-radiotherapy was similar in those with responsive and non-responsive tumours. There were no detectable changes in plasma bFGF through the adenoma-carcinoma sequence or patient groups with non-metastatic cancers. Elevated plasma bFGF was, however, associated with metastatic spread. The significant fall in bFGF in disease-free patients following therapy suggests that bFGF may be useful in clinical practice. This revised version was published online in July 2006 with corrections to the Cover Date.     

GROα in the Fetomaternal and Amniotic Fluid Compartments During Pregnancy and Parturition

PROBLEM: The use of monoclonal antibodies for CD45RA and CD45RO antigens is important in defining maturational and functional stages on lymphocytes. METHOD: To characterize distribution of two isoforms of CD45 antigen CD45RA and CD45RO on CD3⁺, CD4⁺, CD8⁺ and CD56⁺ lymphocyte subsets from first trimester human decidua, two-color flow cytometry were used. RESULTS: In decidua, there were much higher levels of CD45RO^a+ and much lower levels of CD45RA⁺ cells among CD3⁺, CD4⁺, and CD8⁺ cells as compared with peripheral blood samples of the same pregnant women. Only ?40% of CD56⁺ cells in decidua expressed CD45RA. Unlike peripheral blood, ?30% of decidual natural killer (NK) cells weakly stained with anti-CD45RO antibodies. Double-negative CD45RA^? CD45RO^? NK cells were also present in decidua. CONCLUSIONS: The significantly raised percentage of intradecidual T cells expressing CD45RO suggest decidual accumulation of antigen-committed memory cells. The patterns of CD45 isoforms expression on decidual CD56⁺ cells are consistent with hypothesis that uterine CD56⁺ lymphocytes are terminally differentiated cells of NK lineage. PROBLEM: GROα/MGSA is a new member of the chemokine superfamily CXC(α) and is produced by a variety of cells including macrophages, fibroblasts, epithelial, and endothelial cells, and keratinocytes. This chemokine has chemoattractant activity and may participate in neutrophil recruitment and activation during the course of intrauterine infection. This study was conducted to investigate the effect of labor and microbial invasion of the amniotic cavity (MIAC) on amniotic fluid, fetal, and maternal plasma GROα concentrations. METHOD: A cross-sectional study was designed using parameters that included gestational age, results of amniotic fluid (AF) cultures, and labor status at the time of amniocentesis. Fluid was retrieved by transabdominal amniocentesis. MIAC was defined as a positive amniotic fluid culture for bacteria. Umbilical cord blood was retrieved at the time of delivery. Amniotic fluid, maternal and fetal plasma GROα concentrations were measured with a sensitive and specific ELISA (Quantikine, R&D Systems, Minneapolis, MN). RESULTS: 1) GROα was detectable in amniotic fluid, umbilical cord, and maternal plasma samples; 2) GROα concentrations in amniotic fluid increased with advancing gestational age; 3) Both term and preterm gestations with MIAC were associated with higher amniotic fluid GROα concentrations than those with sterile amniotic fluid, independent of the labor status (term, MIAC, labor: median 2.7 ng/ml, range 1.4–12.7 vs. term, no MIAC, labor: median 2.1 ng/ml, range 0.7-3.4, vs term, no MIAC, no labor: median 1.9 ng/ml, range 1.8-4.2; P <0.005; preterm: MIAC median 5 ng/ml, range 0.6–47.9 vs. no MIAC: median 2.3 ng/ml, range 0.5–10; P <0.008); 4) A strong correlation was found between umbilical cord plasma GROα concentrations and neonatal neutrophil count, and between GROα concentrations and white blood cell count in the amniotic fluid (r = 0.67, P < 0.0005 and r ? 0.38, P < 0.001, respectively). CONCLUSION: GROα is a physiologic constituent of amniotic fluid and cord blood. Amniotic fluid GROα concentrations increase with gestational age. Intrauterine infection both preterm and at term is associated with an increase in GROα concentrations of amniotic fluid, suggesting that GROα may play an important role in recruitment of neutrophils into the amniotic cavity.     

Pharmacokinetic studies of high-dose metoclopramide with and without forced diuresis

Summary The pharmacokinetics of high-dose metoclopramide (10 mg/kg body wt. in five infusions of 2 mg/kg body wt. each) was studied in 11 patients (5 females, 6 males) in two groups: group A with and group B (consisting of five patients) without forced diuresis. When the drug was infused, forced diuresis had no influence on the pharmacokinetics of metoclopramide (serum level after the 1st infusion was 851±361 ng/ml in group A versus 840±348 ng/ml in group B; after the 5th infusion it was 2,005±588 ng/ml in group A versus 2,463±1,350 ng/ml in group B). There were significant differences in the 24-h serum levels (582±308 ng/ml in group A versus 379±170 ng/ml in group B;P<0.05) and in the elimination half life (8.5±2.6 h in group A versus 6.1±1.1 h in group B;P<0.05). The results demonstrate that the dosage regimen originally suggested by Gralla for cytostatic drugs, with forced diuresis for high-dose metoclopramide therapy, may also be applied, with no dosage reduction, with to other cytostatic drugs which do not require forced diuresis.     

25-hydroxyvitamin D, 24, 25-dihydroxyvitamin D and 1,25-dihydroxyvitamin D in human cerebrospinal fluid

Summary Samples of CSF and plasma were obtained simultaneously from 46 adult patients who had no endocrine disorders and were undergoing routine diagnostic lumbar puncture because of suspected or proved prolapse of a disc. Concentrations of 25-OHD, 24,25(OH)₂D and 1,25(OH)₂D were measured. The samples were purified by column chromatography and fractionated by HPLC. In the appropriate fractions the vitamin D metabolites were measured by PBA, and cytoreceptor assay. The results were as follows (median, range in brackets): 25-OHD in CSF 8.3 ng/ml (2.0–24.8), in plasma 14.5 ng/ml (7.0–36.0). 24,25(OH)₂D in CSF 1.8 ng/ml (0.3–4.6) and 2.5 ng/ml (0.4–4.7) in plasma. 1.25(OH)₂ D in CSF 25.0 pg/ml (2.2–39.0) and 31.0 pg/ml (10.1–55.0) in plasma. The correlations between plasma and CSF concentrations were as follows: 25-OHDr=0.479 (P<0.001); 24,25(OH)₂Dr=0.815 (P<0.001) and for 1.25(OH)₂Dr=0.497 (P<0.001).Our findings showed vitamin D metabolites to be present in human CSF.Abbreviations Ca Calcium - CSF Cerebrospinal fluid - Vitamin D₃ Cholecalciferol - CPM Counts per min - 24, 25 (OH)₂D 24, 25-dihydroxyvitamin D₃ - 1,25(OH)₂D 1,25-dihydroxyvitamin D₃ - Vitamin D₂ Ergocalciferol - HPLC High-pressure liquid chromatography - 25OHD 25-hydroxyvitamin D₃ - PTH Parathyroid hormone - PBA Protein binding assay - RIA Radioimmunoassay - D-CaBP Vitamin D dependent calcium-binding protein     

Plasma Interleukin-12 Is Elevated in Patients with Preeclampsia

PROBLEM: Abnormal immune activation has been suggested as a contributor to the development of preeclampsia. We hypothesized that intact interleukin (IL)-12 directly, or through its main mediator, interferon (IFN)-γ, contributes to the altered immune response observed in preeclampsia. METHOD OF STUDY: Plasma samples were collected from 20 patients with preeclampsia and 20 normotensive patients with uncomplicated pregnancies who were matched with the preeclamptic patients by age, gestational age, and parity. Samples were collected before the onset of labor, induction, or medical intervention. The samples were assayed for IL-12 and IFN-γ by specific enzyme-linked immunoassays. RESULTS: IL-12 was detected in 35% of the preeclamptic patients and in 5% of the patients with normal pregnancies (P < 0.01). The detection rate and mean concentration of IFN-γ were comparable in both groups. CONCLUSION: Intact plasma IL-12 is detected more frequently in preeclamptic patients, suggesting the involvement of this cytokine in the enhanced immune response observed in preeclampsia.     

Urinary vascular endothelial growth factor concentrations in women undergoing gonadotrophin treatment

A recently identified cytokine, vascular endothelial growthfactor (VEGF, vascular permeability factor) has been implicatedin ovarian hyperstimulation syndrome in women undergoing assistedreproduction. We postulate that circulating and urinary VEGFvalues increase following gonadotrophin stimulation, in parallelwith the increased ovarian vascularity. A VEGF radioimmunoassaywas developed using iodinated VEGF as tracer, a goat anti-VEGFserum as antiserum and recombinant human VEGF as standard. Thespecificity of the assay was confirmed by comparing the reversephase high-performance liquid chromatography (HPLC) patternof VEGF immunoactivity in urine and urine spiked with recombinantVEGF. Urine was concentrated 5-fold prior to measurement bythe radioimmunoassay. VEGF:creatinine ratios in early morningurine samples were used to monitor daily urinary VEGF concentrationsbased on its high correlation (r = 0.77, P < 0.001) withVEGF concentrations in 24 h urine collections. No diurnal variationin VEGF:creatinine ratios was detected. VEGF:creatinine ratioswere determined daily from nine women undergoing gonadotrophin-releasinghormone (GnRH) agonist/gonadotrophin treatment. In a further16 women, early morning urine samples were collected in theperi-ovulatory period. A significant increase (P < 0.005,n = 25) was observed in VEGF:creatinine ratios following humanchorionic gonadotrophin (HCG) administration. VEGF:creatinineratios correlated poorly ((r < 0.34) with plasma oestradiol,follicle number and size. It is concluded that urinary VEGF/creatinineratios increase following HCG stimulation. assisted reproduction/gonadotrophins/in-vitro fertilization/ovarian hyperstimulation syndrome/vascular permeability factor     

Endothelin-1 immunoreactivity in plasma is elevated in HIV-1 infected patients with retinal microangiopathic syndrome

Endothelin-1 is a recently identified cytokine with potent vasoconstrictor activity which is associated with various diseases involving blood vessels. HIV-1 related retinal microangiopathic syndrome is a frequent finding in patients with AIDS or AIDS-related complex, presenting predominantly with retinal cotton-wool spots. We investigated 55 HIV-1 infected patients by ophthalmoscopy and for endothelin-I immunoreactivity in plasma and an additional 76 HIV-1 infected patients only for endothelin-1 levels. For reference values 13 age-matched healthy subjects were studied. In 18 of 55 patients (33%) investigated ophthalmoscopically we found evidence of microangiopathic syndrome. Overall, the mean endothelin-1 immunoreactivity in plasma of HIV-1 infected patients was significantly elevated as compared to controls (4.28 ± 3.62 versus 2.72 ± 0.67 fmol/ml, P < 0.0001). HIV-1 infected patients with retinal microangiopathic syndrome had significantly higher plasma levels of endothelin-1 immunoreactivity (4.59 ± 1.38 fmol/ ml) compared to HIV-1 infected patients without microangiopathic syndrome (3.18 ± 1.64 fmol/ml, P = 0.003). Correlation analysis revealed that endothelin-1 immunoreactivity in plasma had no significant association with disease progression, CD4 cell count, ₂-mi-croglobulin, neopterin, or age. Endothelin-1 immunoreactivity in plasma was correlated exclusively with retinal microangiopathic syndrome in one or both eyes (r = 0.45, P = 0.0006) and with the number of cotton-wool spots (r = 0.50, P = 0.0001). In conlusion, HIV-1 related retinal microangiopathic syndrome is associated with elevated plasma levels of endothelin-1. By virtue of its potent vasoconstrictor activity endothelin-1 may be involved in the pathogenesis of HIV-1 related vascular disease.Abbreviations AIDS aquired immunodeficiency syndrome - ET-1 endothelin-1 - HIV human immunodeficiency virus - IR immunoreactivity - WR Walter Reed classification Correspondence to: B. Rolinski     

Plasma soluble endothelial selectin is elevated in women with pre- eclampsia

The study was conducted to determine whether altered plasma concentrations of soluble selectins are involved in the pathogenesis of pre-eclampsia. Maternal plasma samples were collected from 20 patients with pre-eclampsia, and from 20 matched normotensive patients with uncomplicated pregnancies. Samples were assayed for soluble endothelial selectin (sES), platelet selectin (sPS) and leukocyte selectin (sLS) by specific enzyme-linked immunosorbent assay. The three soluble selectins were detectable in the plasma of all pre-eclamptic and control patients. The mean plasma concentrations of sPS and sLS were comparable between the groups. However, the mean plasma concentration of sES was significantly higher in the pre-eclamptic group compared with the control group (61 ng/ml +/- 30 ng/ml compared with 40 ng/ml +/- 17 ng/ml; P < 0.01). The selective increased plasma concentrations of sES in patients with pre-eclampsia provide specific evidence for endothelial activation and may reflect distinct pathways for neutrophil activation in pre-eclampsia.      

Changes of placental syndecan-1 expression in preeclampsia and HELLP syndrome

Preeclampsia is characterized by maternal systemic anti-angiogenic and pro-inflammatory states. Syndecan-1 is a cell surface proteoglycan expressed by the syncytiotrophoblast, which plays an important role in angiogenesis and resolution of inflammation. Our aim was to examine placental syndecan-1 expression in preeclampsia with or without hemolysis, elevated liver enzymes, and low platelet count (HELLP) syndrome. Placentas were obtained from women in the following groups: (1) late-onset preeclampsia (n?=?8); (2) early-onset preeclampsia without (n?=?7) and (3) with HELLP syndrome (n?=?8); (4) preterm controls (n?=?5); and (5) term controls (n?=?9). Tissue microarrays (TMAs) were constructed from paraffin-embedded placentas. TMA slides were immunostained for syndecan-1 and evaluated using microscopy, virtual microscopy, and semi-automated image analysis. Maternal sera from patients with preeclampsia (n?=?49) and controls (n?=?32) were immunoassayed for syndecan-1. BeWo cells were treated with Forskolin or Latrunculin B or kept in ischemic conditions. SDC1 expression and syndecan-1 production were investigated with qRT-PCR, confocal microscopy, and immunoassays. Syndecan-1 was localized to the syncytiotrophoblast apical membrane in normal placentas. Syndecan-1 immunoscores were higher in late-onset preeclampsia (p?=?0.0001) and early-onset preeclampsia with or without HELLP syndrome (p?=?0.02 for both) than in controls. Maternal serum syndecan-1 concentration was lower in preeclampsia (median, 673 ng/ml; interquartile range, 459–1,161 ng/ml) than in controls (1,158 ng/ml; 622–1,480 ng/ml). SDC1 expression and syndecan-1 immunostainings in BeWo cells and syndecan-1 concentrations in supernatants increased during cell differentiation. Disruption of the actin cytoskeleton with Latrunculin B decreased syndecan-1 release, while ischemic conditions increased it. Syncytiotrophoblastic syndecan-1 expression depends on the differentiation of villous trophoblasts, and trophoblastic syndecan-1 release is decreased in preeclampsia and HELLP syndrome. This phenomenon may be related to the disturbed syncytiotrophoblastic cortical actin cytoskeleton and associated with maternal anti-angiogenic and pro-inflammatory states in these syndromes.     

A selective increase in plasma soluble vascular cell adhesion molecule-1 levels in preeclampsia.

PROBLEM: The study was conducted to determine whether altered plasma levels of soluble intercellular adhesion molecule (ICAM)-1 and soluble vascular cell adhesion molecule (VCAM)-1 are involved in the pathogenesis of preeclampsia. METHOD OF STUDY: Maternal plasma samples were collected from 20 patients with preeclampsia, 20 matched normotensive patients with uncomplicated pregnancies. and ten healthy nonpregnant women. Samples were assayed for soluble VCAM-1 and soluble ICAM-1 by specific enzyme-linked immunosorbent assay. RESULTS: Both soluble VCAM-1 and soluble ICAM-1 were detectable in the plasma of all preeclamptic, normotensive pregnant, and nonpregnant women. The mean plasma level of soluble VCAM-1 was significantly higher in preeclamptic women compared to normotensive pregnant women (1831 ng/mL +/- 534 ng/mL vs. 1254 ng/mL +/- 386 ng/mL, respectively; P < 0.05). However, the plasma level of soluble VCAM-1 was unchanged during the third-trimester of normal pregnancy compared to nonpregnant women. The mean plasma level of soluble ICAM-1 in preeclamptic and normotensive pregnant women were increased when compared to nonpregnant women. However, the mean plasma level of soluble ICAM-1 was comparable in women with preeclampsia and normotensive pregnancy. CONCLUSIONS: The selective increased plasma levels of soluble VCAM-1 in patients with preeclampsia provide evidence for endothelial activation and suggest distinct pathways for neutrophil and endothelial activation in preeclampsia.     

Validation of a sensitive and specific real‐time PCR for detection and quantitation of hepatitis B virus covalently closed circular DNA in plasma of chronic hepatitis B patients

Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) serves as a template for viral replication and plays a role in persistence of HBV infection. The origin and significance of cccDNA in plasma however, is not well understood. A sensitive, specific, and reproducible real‐time PCR for detection and quantitation of cccDNA in plasma of chronic hepatitis B patients was developed and validated. Four HBV DNA reference panels, and 96 plasma samples of chronic hepatitis B patients were analyzed. Results were compared with total HBV DNA levels, individual ALT levels and the Histology Activity Index (HAI). This cccDNA assay had a lower limit of detection at 15 copies/PCR, a lower limit of quantitation at 91 copies/PCR and a correlation coefficient (R) of 0.98 (P < 0.0001). cccDNA was detected in two of four international panels. Significant correlation was found between cccDNA and total HBV DNA levels in both panels (R = 0.96, and R = 0.43) and in samples of the chronic hepatitis B patients (R = 0.88, P < 0.0001). In 57% of these samples cccDNA was detectable. Mean level of cccDNA was 0.16% of total HBV load. Plasma cccDNA levels were higher in HBeAg positive samples than in HBeAg negative samples (4.91 log copies/ml vs. 3.88 log copies/ml, P < 0.0001). Levels of total HBV DNA and HBV genotype did not influence cccDNA detection. ALT levels and HAI‐score were not correlated with plasma cccDNA levels. These findings suggest that cccDNA levels in plasma are not the result of increased hepatocyte degeneration, but indicate that other mechanisms might be responsible. J. Med. Virol. 81:988–995, 2009. © 2009 Wiley‐Liss, Inc.     

Macrophage Inflammatory Protein-1α in Term and Preterm Parturtition: Effect of Microbial Invasion of the Amniotic Cavity

PROBLEM: This study was conducted to determine whether: (1) gestational age, parturition, and microbial invasion of the amniotic cavity (MIAC) are associated with changes in amniotic fluid concentrations of immunoreactive macrophage inflammatory protein-1α; (2) amniotic fluid concentrations of macrophage inflammatory protein-1α are correlated with the white blood cell count and the concentrations of interleukin-8 in amniotic fluid. METHOD: Amniotic fluid was retrieved by amniocentesis from 126 patients; 54 women with preterm labor and intact membranes (no MIAC-delivery at term, N = 21; no MIAC-preterm delivery, N = 16; MIAC-preterm delivery, N = 17); 62 patients at term (no labor, N = 19; labor-no MIAC, N = 20; labor-MIAC, N = 23); and 10 patients in the midtrimester of pregnancy. Amniotic fluid was cultured for aerobic, anaerobic and Mycoplasma species. Determinations of amniotic fluid macrophage inflammatory protein-1α and interleukin-8 were performed with immunoassays validated for amniotic fluid (sensitivity: 14.2 pg/ml and 0.3 ng/ml, respectively). Kruskal-Wallis analysis of variance (ANOVA) for censored data, Mann-Whitney U test and Spearman's rank correlation were performed for analysis. RESULTS: 1) Amniotic fluid macrophage inflammatory protein-1α was present in only 31.0% (9/29) of patients not in labor (midtrimester and term). 2) Patients with preterm labor and MIAC had higher amniotic fluid concentrations of macrophage inflammatory protein-1α than those without MIAC (no MIAC-delivery at term: median 0.0 pg/ml, range 0.0–221.2; no MIAC-preterm delivery: median 37.4 pg/ml, range 0.0–494.6; MIAC-preterm delivery: median 7171.0 pg/ml, range 402.5–37994.0; P<0.00001). 3) Among patients at term, MIAC was associated with higher concentrations of amniotic fluid macrophage inflammatory protein-1α than patients without MIAC (no labor: median 0.0 pg/ml, range 0.0–25.6; labor-no MIAC: median 16.7 pg/ml, range 0.0–161.6; labor-MIAC: median 103.8 pg/ ml, range 0.0–4349.0, P<0.001). 4) Among patients in preterm labor, a strong correlation was found between amniotic fluid concentrations of macrophage inflammatory protein-1α and interleukin-8 (r = 0.9, P<0.00001) and between amniotic fluid macrophage inflammatory protein-1α concentrations and amniotic fluid white blood cell count (r = 0.6, P<0.0001). CONCLUSIONS: (1) Macrophage inflammatory protein-1α is undetectable in most amniotic fluid samples from patients in the midtrimester of pregnancy and at term not in labor. (2) Microbial invasion of the amniotic cavity is associated with increased concentrations of immunoreactive amniotic fluid macrophage inflammatory protein-1α in both term and preterm gestations. (3) Amniotic fluid macrophage inflammatory protein-1α concentrations significantly correlate with interleukin-8 levels and white blood cell count in amniotic fluid. Our data strongly suggest a role for macrophage inflammatory protein-1α in the mechanisms responsible for the recruitment of leukocytes into the amniotic cavity during the course of intrauterine infection.     

FcγRIIB‐nt645+25A/G gene polymorphism and periodontitis in Japanese women with preeclampsia

FcγRIIB contains a unique immunoreceptor tyrosine‐based inhibition motif (ITIM) and functions as a negative feedback regulator of leucocyte activation and antibody production. We have previously reported FcγRIIB‐nt645+25A/G gene polymorphism to be associated with prevalence and severity of periodontitis, FcγRIIB expression level on peripheral B lymphocytes and the serum IgG level against periodontopathic bacteria. Previous studies have reported maternal periodontal disease to be associated with an increased risk for preeclampsia. Therefore, FcγRIIB‐nt645+25A/G gene polymorphism may be associated with preeclampsia by affecting immune response to periodontopathic bacteria in pregnant women. To elucidate whether FcγRIIB‐nt645+25A/G gene polymorphism has associations with preeclampsia and/or periodontitis in pregnant Japanese women, a case–control study was carried out on women with preeclampsia (n = 13) and without preeclampsia (n = 106). Maternal periodontal parameters and bacterial data of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in subgingival plaque were collected within 5 days of delivery. FcγR genotypes of each woman were determined using the genomic DNA isolated from peripheral blood. Serum IgG levels specific for each bacteria were determined. There was a significant association between FcγRIIB‐nt645+25A/G polymorphism and preeclampsia (P = 0.013). The frequency of the FcγRIIB‐nt645+25AA genotype was higher in the preeclampsia group compared with the nonpreeclampsia group (P = 0.007). The DNA level of A. actinomycetemcomitans from subgingival plaque was shown to be higher in the preeclampsia group (P = 0.017). In conclusion, maternal FcγRIIB‐nt645+25A/G polymorphism and subgingival DNA level of A. actinomycetemcomitans were significantly associated with the prevalence of preeclampsia in a limited number of Japanese women independently with periodontal infection. Further investigations should be performed to confirm this association in a larger population and to determine the biological process of the association.     

Relationship between serum complement and different lipid disorders

Inflammatory and lipid factors share an important role in atherosclerosis. Recent studies showed the concomitant presence and increase of complement components and lipid both in the atherosclerotic plaque and the circulating blood. The aim of this study was to examine the relationship between the complement system and lipid disorders. We evaluated the circulating complement terminal complex C5b-9, a clear sign of complement activation, in three groups of 30 patients the first with hypercholesterolemia, the second with hypertriglyceridemia (associated with low values of HDL-cholesterol), the third with low levels of HDL-cholesterol compared with an equivalent group of matched normolipemic subjects. We found a significant increase of sC5b-9 in each group of patients compared with controls. The mean sC5b-9 level in the hypercholesterolemic population was 366.2±141.2 ng/ml (P<0.01), 395.4±118.2 ng/ml in the hypertrygliceridemic group (P<0.01), 414.8±126.4 ng/ml in the low HDL-chol subjects (P<0.01), and 182.0±40.8. ng/ml in the control group. Regression analysis showed a significant direct correlation between sC5b-9 and triglycerides (r=0.64), and a significant inverse correlation between sC5b-9, HDL-chol (r=0.74), and apo-A1 (r=0.68); no significant relationship was found between sC5b-9 and cholesterol. We suggest that complement activation is associated with the various lipid disorders and is more important in those dyslipidemic conditions in which other factors may be involved. In particular, hypertriglyceridemia may be associated with endothelial and fibrinolytic disturbances, and the decrease of HDL may induce the failure of the regulatory proteins transported by the same HDL. Received: 31 March 2001 / Accepted: 3 February 2002     

Hormonal responses to training and its tapering off in competitive swimmers: relationships with performance

During a winter training season, the effects of 12 weeks of intense training and 4 weeks of tapering off (taper) on plasma hormone concentrations and competition performance were investigated in a group of highly trained swimmers (n = 8). Blood samples were collected and the swimmers performed their speciality in competition at weeks 10 (mid-season), 22 (pre-taper) and 26 (post-taper). No statistically significant changes were observed in the concentrations of total testosterone (TT), non-sex hormone binding globulin-boundtestosterone (NSBT), cortisol (C), luteinising hormone, thyroid stimulating hormone, triiodothyronine, thyroxine plasma catecholamines, creatine kinase and ammonia during training and taper. Mid-season NSBT: C ratio and the amount of training were statistically related (r = 0.82,P < 0.05). Competition performance slightly declined during intense training [0.52 (SD 2.51) %, NS] and improved during taper [2.32 (SD 1.69)%,P < 0.01]. Changes in performance during training and taper correlated with changes in ratios TT: C (r = 0.86,P < 0.01andr = 0.81,P < 0.05, respectively) and NSBT: C (r = 0.77,P < 0.05 andr = 0.76,P < 0.05, respectively). In summary, these results showed that the monitored plasma hormones and metabolic indices were unaltered by 12 weeks of intense training and 4 weeks of taper. The TT: C and NSBT: C ratios, however, appeared to be effective markers of the swimmers' performance capacities throughout the training season.     

Elevated procalcitonin as a diagnostic marker in meningococcal disease

Patients with meningococcal disease who seek medical attention can create a diagnostic dilemma for clinicians due to the nonspecific nature of the disease’s presentation. This study assesses the diagnostic accuracy of procalcitonin levels in the setting of meningococcal disease. Two emergency department cohorts (A and B) were studied between 2002 and 2005, during the current epidemic of serogroup B meningococcal disease in New Zealand. Cohort A consisted of 171 patients, all with confirmed meningococcal disease (84 children, 87 adults). Cohort B consisted of a large (n=1,524) consecutively recruited population of febrile patients who presented to the emergency department, 28 of whom had confirmed meningococcal disease. Within the meningococcal disease cohort (cohort A), the geometric mean procalcitonin level was 9.9 ng/ml, with levels being higher in children than in adults (21.6 vs. 4.6 ng/ml, p=0.01). The overall sensitivity of elevated procalcitonin, using a cutoff of 2.0 ng/ml in children and 0.5 ng/ml in adults, was 0.93 (95%CI: 0.88–0.96). Despite the higher cutoff level for paediatric patients, a trend towards greater sensitivity existed in children (0.96 vs. 0.90; p=0.08). Elevated procalcitonin was correlated with whole blood meningococcal load (r=0.50) and Glasgow Meningococcal Sepsis Prognostic Score (r=0.40). Within the cohort of patients who were febrile on presentation (cohort B), the specificity of elevated procalcitonin in meningococcal disease was 0.85 (95% CI: 0.83–0.87), the positive and negative likelihood ratios were 6.1 and 0.08, respectively, and the sensitivity of elevated procalcitonin (0.93; 95% CI: 0.76–0.99) was corroborated. Measurement of procalcitonin is a useful tool in patients with nonspecific febrile illnesses when the possibility of meningococcal disease is present. The diagnostic accuracy surpasses that of current early laboratory markers, allowing results to be used to guide decisions about patient management.     

ESTABLISHMENT OF A HIGHLY SENSITIVE LEPTIN RADIOIMMUNOASSAY AND DETECTION OF INCREASED LEPTIN LEVELS IN HYPERLIPIDEMIA AND PREGNANCY

ABSTRACT

The highly effective antibody has been obtained by immunizing rabbits with recombinant leptin many times. The leptin is iodinated with the chloramine-T method and purified with a Sephadex-G25 chromatography column. The reaction between antigen and antibody is carried out by a one-step balance method and cultured at 4°C for 24 h; the binding and free antigen was then separated by PR reagent. The determining range of this method is about 0.5–24 ng/mL; limited detection level is 0.45 ng/mL, relative standard deviation in a group, and among groups, are less than 5.4% and 8%, respectively. The level of blood leptin in 277 samples of normal persons, in 112 samples of overweight persons (weight/hieght m² ≥ 25) and 224 samples of hyperlipidemic patients have been measured by this method. It is demonstrated that the level of blood leptin in males is much lower than that of the females, and becomes elevated with increased age. Serum leptin level in overweight persons and hyperlipidemic patients is also much higher than that of normal groups (P<0.01). Serum leptin of 21 workers in our lab at 8:00AM and 4:00PM has been tested. It was found that there are no differences between the two time points. The same results are obtained within age groups. Leptin levels of pregnant women's serum is higher than those of the control group (P<0.001). Leptin in newborn's serum is significantly lower than those of mothers (P<0.01). There is no obvious correlation between leptin level of mother and newborns by correlation analysis (r = 0.19, P>0.05). The body weight and body weight index of pregnant women are well correlated with their serum leptin levels (r = 0.33 and 0.35, P<0.05). The body weight and body weight index of newborns are well correlated with their serum leptin levels (r = 0.54 and 0.49, P<0.001). The serum leptin level of pregnant women is not correlated with newborn's body weight (r = 0.10). These results have shown that the proposed method is stable, simple, and specific, being sensitive enough to determine leptin levels in human serum or plasma.