Amniotic Fluid Matrix Research Articles

The relationship between the intensity of intra-amniotic inflammation and the presence and severity of acute histologic chorioamnionitis in preterm gestation

Objective: Acute histologic chorioamnionitis (HCA) is associated with an increased risk of perinatal mortality and morbidity. The purpose of this study was to determine the relationship between the intensity of intra-amniotic inflammation (IAI) and the severity of acute HCA in preterm gestation.Methods: The relationship between the intensity of IAI and the presence and severity of acute HCA was examined in 412 patients with singleton gestations who delivered within 120 h of transabdominal amniocentesis. The concentration of amniotic fluid (AF) matrix metalloproteinase (MMP)-8 was assayed to determine the presence and intensity of IAI. Acute HCA was defined as the presence of inflammatory change in any tissue samples according to the criteria previously reported. The total grade of acute HCA was used to determine the severity of HCA.Results: (1) Patients with IAI had a significantly higher rate of acute HCA than those without IAI [76.9% (133/173)] versus 20.9% (50/239), p < 0.001]. The AF MMP-8 concentration was significantly higher in patients with acute HCA than in those without acute HCA (median [range]; 188.3 ng/ml [0.3–6142.6] versus 1.8 ng/ml [0.3–2845.5], p < 0.001); (2) Of 183 patients with acute HCA, the AF MMP-8 concentration was positively correlated with the severity of acute HCA (p < 0.001).Conclusions: AF MMP-8 concentration was not only a predictor of the presence of acute HCA, but its concentration also correlated with the severity of acute HCA. The higher the intensity of IAI, the worse the degree of acute HCA in preterm gestation.

The assessment of the relationship between amniotic fluid matrix metalloproteinase-9 and zinc levels with adverse obstetric outcomes

To examine the relationship between the amniotic fluid MMP-9 and zinc levels during 16-19th gestational weeks and perinatal outcomes. One hundred and seventeen singleton pregnancies that underwent genetic amniocentesis from January 2005 through November 2009 were evaluated. Subjects were divided into two main groups: a control group (group 1) (n: 74), and an adverse obstetric outcomes group (group 2) (n: 43). Group 2 consisted of the following: preterm birth group, gestational hypertension and preeclampsia group, gestational diabetes group, fetal growth restriction group, macrosomia group, and pregnancy loss group. MMP-9 and zinc (Zn) values in the amniocentesis materials sampled between the 16th and 19th gestational weeks were analyzed retrospectively in terms of perinatal outcomes. Any significant difference among the groups was assessed by unpaired samples t test and the Mann-Whitney U test. Statistical significance was defined as p < 0.05. A comparison among groups showed no significant difference in terms of Zn results between the group 1 and 2 (p = 0.879). MMP-9 levels were significantly lower in both the preterm birth group (p = 0.043) and group 1 (p = 0.015). We found that the amniotic fluid MMP-9 levels of patients who delivered preterm were significantly lower between the 16th and 19th gestational weeks.

Fetal, amniotic and maternal inflammatory responses in early stage of ascending intrauterine infection, inflammation restricted to chorio-decidua, in preterm gestation

Objective: No data exist on the frequency and intensity of the fetal, intraamniotic and maternal inflammation in preterm-gestations with inflammation restricted to chorio-decidua, early stage of ascending intrauterine infection. The objective of the study is to examine this issue.Study Design: The frequency and intensity of fetal (cord blood C-reactive protein [CRP] at birth >200 ng/ml), intraamniotic (amniotic fluid matrix metalloproteinase-8 [MMP-8] >23 ng/ml) and maternal (maternal serum CRP >0.7 ng/ml) inflammation were compared in 304 singleton preterm-gestations (<35.4 weeks) delivered within 5 days of amniocentesis. Placental pathology was divided into placenta without any inflammation, inflammation restricted to chorio-decidua and inflammation beyond chorio-decidua.Results: Intraamniotic inflammation, but not fetal or maternal inflammation, was significantly more frequent (43.2% versus 10.5%; p < 0.005) and intense (median amniotic fluid MMP-8; 7.5 ng/ml versus 1.3 ng/ml; p < 0.001) in inflammation restricted to chorio-decidua than in placenta without any inflammation. However, inflammation restriced to chorio-decidua was associated with a significantly lower rate and intensity of fetal, intraamniotic and maternal inflammation than inflammation beyond chorio-decidua (each for p < 0.05).Conclusion: Intraamniotic inflammation was more frequent and intense in patients with inflammation restricted to chorio-decidua than in those without placental inflammation. However, fetal and maternal inflammatory responses were similar between these two groups.

An elevated maternal serum C-reactive protein in the context of intra-amniotic inflammation is an indicator that the development of amnionitis, an intense fetal and AF inflammatory response are likely in patients with preterm labor: clinical implications

sObjective: We propose that an elevated maternal serum C-reactive protein (CRP) concentration in the context of intra-amniotic inflammation (IAI) is a predictor for amnionitis development, known to be the most advanced stage of maternal inflammatory response during the progression of acute histologic chorioamnionitis in preterm gestations.Methods: Study population consisted of 53 singleton gestations with IAI, who underwent amniocentesis due to preterm labor and intact membranes (PTL) and delivered preterm-neonates (<34.5 weeks) within 5 days of amniocentesis. The frequency of amnionitis and the intensity of fetal and amniotic fluid (AF) inflammatory response were examined according to the presence or absence of an elevated maternal serum CRP (≥0.7 mg/dL) at the time of amniocentesis. IAI was defined as an elevated AF matrix metalloproteinase-8 (MMP-8) (≥23 ng/mL), and fetal inflammatory response syndrome (FIRS) defined as an elevated umbilical cord plasma CRP (≥200 ng/mL).Results: (1) Patients (73.6%, 39/53) with an elevated maternal serum CRP had a significantly higher rate of amnionitis (59.0% versus 7.1%; p < 0.005), but not funisitis (46.2% versus 28.6%; p > 0.05), and higher median AF MMP-8 and umbilical cord plasma CRP concentration at birth than patients (26.4%,14/53) without that (AF MMP-8 (ng/mL): 373.1 versus 138.6: p = 0.05; umbilical cord plasma CRP (ng/mL): 363.4 versus 15.5: p < 0.05); (2) Multiple logistic regression analysis demonstrated that an elevated maternal serum CRP was a better independent predictor of amnionitis (odds ratio (OR), 12.5: 95% confidence interval (CI), 1.1–141.0; p < 0.05) than FIRS (OR, 3.6: 95% CI, 0.6–20.2; p = 0.150) and any other AF tests.Conclusions: An elevated maternal serum CRP concentration in the context of IAI is an indicator that the development of amnionitis, an intense fetal and AF inflammatory response are likely in patients with PTL.

The frequency and clinical significance of intra-amniotic inflammation defined as an elevated amniotic fluid matrix metalloproteinase-8 in patients with preterm labor and low amniotic fluid white blood cell counts

ObjectiveTo determine the frequency and clinical significance of intra-amniotic inflammation (IAI) defined as an elevated amniotic fluid (AF) matrix metalloproteinase-8 (MMP-8) concentration in patients with preterm labor and intact membranes (PTL) and low AF white blood cell (WBC) counts.MethodsAdverse pregnancy outcomes were compared according to the presence or absence of IAI in 220 singleton gestations who underwent amniocentesis due to PTL (gestational age<35.7 weeks) and had low AF WBC counts (<19 cells/mm3). Adverse pregnancy outcomes included preterm birth within 5 days of amniocentesis, acute histologic chorioamnionitis (acute-HCA) and positive AF culture. IAI was defined as an elevated AF MMP-8 concentration (≥23 ng/mL).ResultsIAI was present in 19% of study population. Adverse pregnancy outcomes were significantly more frequent in patients with IAI than in those without IAI (preterm birth within 5 days of amniocentesis, 88% vs. 41%; acute-HCA, 47% vs. 11%; positive AF culture, 10% vs. 2%; each for P<0.05). Patients with IAI had a significantly shorter median amniocentesis-to-delivery interval than those without IAI (7.8 hours [0.01-3,307.3 hours] vs. 310.3 hours [0.01-2,973.8 hours]; P<0.001 from survival analysis). Multiple logistic regression analysis demonstrated that only an IAI (odds ratio, 3.3; 95% confidence interval, 1.5-7.3; P<0.005) retained a statistical significance in the prediction of acute-HCA after other confounding variables were adjusted.ConclusionApproximately one-fifth of patients with PTL and low AF WBC counts have an evidence of IAI and are at risk for impending preterm delivery and acute-HCA when AF MMP-8 concentration is used.

Which is more important for the intensity of intra-amniotic inflammation between total grade or involved anatomical region in preterm gestations with acute histologic chorioamnionitis?

ObjectiveThere is no data on which is more important for the intensity of intra-amniotic inflammation (IAI) between total grade or involved anatomical region in acute histologic chorioamnionitis (acute-HCA) of preterm-gestations. The objective of current study is to examine this issue.MethodsThe intensity of IAI was measured by amniotic fluid (AF) white blood cell (WBC) count and matrix metalloproteinase-8 (MMP-8) concentration in 225 singleton preterm-gestations (<36 weeks) who had acute-HCA including chorio-decidua involvement and delivered within 5 days of amniocentesis. Acute-HCA was defined in the presence of acute inflammatory changes in each anatomical region (i.e., chorio-decidua, amnion or chorionic plate). Patients were divided into 6 groups according to total grade (i.e., 1-6) and the presence or absence of chorio-decidua restriction (i.e., chorio-decidua restriction vs. extension beyond chorio-decidua) of acute-HCA.ResultsThere was no significant difference in a median AF WBC and MMP-8 between the two groups (group-1, cases with total grade 1 vs. group-2, cases with total grade 2) among cases with chorio-decidua restriction (each for P>0.05) and between the four groups (group-3, cases with total grade 2 vs. group-4, cases with total grade 3 vs. group-5, cases with total grade 4 vs. group-6, cases with total grade 5-6) among cases with extension beyond chorio-decidua (each for P>0.05). However, group-3 (cases with extension beyond chorio-decidua) had a significantly higher median AF WBC and MMP-8 than group-2 (cases with chorio-decidua restriction) among cases with total grade 2 (each for P<0.05).ConclusionInvolved anatomical region is more important than total grade for the intensity of IAI in acute-HCA of preterm-gestations.

Predictive value of intra-amniotic and serum markers for inflammatory lesions of preterm placenta

Objective To compare the relative predictive values of amniotic fluid (AF) matrix metalloproteinase-9 (MMP-9), interleukin-6 (IL-6), and serum C-reactive protein (CRP) for histologic chorioamnionitis and intra-amniotic infection in women with preterm labor or preterm premature rupture of membranes (PROM). Study design This retrospective cohort study included 99 consecutive women with preterm labor or preterm PROM (21–35 weeks’ gestation) who delivered within 72 h of transabdominal amniocentesis. The AF was cultured for aerobic and anaerobic bacteria and for genital mycoplasmas and was assayed for MMP-9 and IL-6 levels. Maternal serum CRP was measured immediately after amniocentesis. The placentas were examined histologically. Main outcome measures histologic chorioamnionitis and intra-amniotic infection. Results The prevalence of histologic chorioamnionitis and a positive AF culture was 44% (44/99) and 28% (28/99), respectively. In predicting intra-amniotic infection, AF MMP-9 had a significantly higher area under the curve (AUC: 0.94 [95% CI, 0.87–0.98]) than AF IL-6 (0.87 [95% CI, 0.78–0.84]; P < 0.05) and serum CRP (0.76 [95% CI, 0.66–0.84]; P < 0.001) and a higher sensitivity and specificity than serum CRP ( P < 0.01, respectively). However, in predicting histologic chorioamnionitis, there were no significant differences in AUCs among the three tests (AF MMP-9: 0.78 [95% CI, 0.68–0.85]; AF IL-6: 0.76 [95% CI, 0.66–0.84]; serum CRP: 0.76 [95% CI, 0.66–0.84]). In a sub-analysis of 71 women without intra-amniotic infection, histologic chorioamnionitis was associated with an elevated serum CRP level ( P < 0.05), but not with the level of AF IL-6 or MMP-9 ( P = 0.232 and P = 0.402, respectively). Conclusions The AF MMP-9 has a better overall diagnostic performance than the AF IL-6 and maternal serum CRP in predicting intra-amniotic infection. However, the serum CRP level obtained up to 72 h before delivery appears to be an important marker for early identification of histologic chorioamnionitis in women without intra-amniotic infection.

Isolation, proliferation, cytogenetic, and molecular characterization and in vitro differentiation potency of canine stem cells from foetal adnexa: A comparative study of amniotic fluid, amnion, and umbilical cord matrix

The possibility to isolate canine mesenchymal stem cells (MSCs) from foetal adnexa is interesting since several canine genetic disorders are reported to resemble similar dysfunctions in humans. In this study, we successfully isolated, cytogenetically and molecularly characterized, and followed the differentiation potency of canine MSCs from foetal adnexa, such as amniotic fluid (AF), amniotic membrane (AM), and umbilical cord matrix (UCM). In the three types of cell lines, the morphology of proliferating cells typically appeared fibroblast-like, and the population doubling time (DT) significantly increased with passage number. For AF- and AM-MSCs, cell viability did not change with passages. In UCM-MSCs, cell viability remained at approximately constant levels up to P6 and significantly decreased from P7 (P < 0.05). Amnion and UCM-MSCs expressed embryonic and MSC markers, such as Oct-4 CD44, CD184, and CD29, whereas AF-MSCs expressed Oct-4, CD44. Expression of the hematopoietic markers CD34 and CD45 was not found. Dog leucocyte antigens (DLA-DRA1 and DLA-79) were expressed only in AF-MSCs at P1. Isolated cells of the three cell lines at P3 showed multipotent capacity, and differentiated in vitro into neurocyte, adipocyte, osteocyte, and chondrocyte, as demonstrated by specific stains and expression of molecular markers. Cells at P4 showed normal chromosomal number, structure, and telomerase activity. These results demonstrate that, in dog, MSCs can be successfully isolated from foetal adnexa and grown in vitro. Their proven stemness and chromosomal stability indicated that MSCs could be used as a model to study stem cell biology and have an application in therapeutic programs.

Comparison of equine bone marrow-, umbilical cord matrix and amniotic fluid-derived progenitor cells

The aim of the study was to compare in vitro the stemness features of horse progenitor cells derived from bone marrow (BM-MSCs), amniotic fluid (AF-MSCs) and umbilical cord matrix (EUC-MSCs). It has been suggested that there may be a stem cell population within both umbilical cord matrix and amniotic fluid. However, little knowledge exists about the characteristics of these progenitor cells within these sources in the equine species. This study wanted to investigate an alternative and non-invasive stem cell source for the equine tissue engineering and to learn more about the properties of these cells for future cell banking. Bone marrow, umbilical cord and amniotic fluid samples were harvested from different horses. Cells were analyzed for proliferation, immunocytochemical, stem cell gene expression and multilineage plasticity. BM- and AF-MSCs took similar time to reach confluence and showed comparable plating efficiency. All cell lines expressed identical stem cell markers and capability to differentiate towards osteogenic lineage. Almost all cell lines differentiated into the adipogenic lineage as demonstrated by cytochemical staining, even if no adipose gene expression was detectable for AF-MSCs. AF- and EUC-MSCs showed a limited chondrogenic differentiation compared with BM-MSCs as demonstrated by histological and biochemical analyses. These findings suggest that AF-MSCs appeared to be a readily obtainable and highly proliferative cell line from an uninvasive source that may represent a good model system for stem cell biology. More studies are needed to investigate their multilineage potential. EUC-MSCs need to be further investigated regarding their particular behavior in vitro represented by spheroid formation.

312 ISOLATION, PROLIFERATION, AND CHARACTERIZATION OF MESENCHYMAL STEM CELLS FROM AMNIOTIC FLUID, AMNION, AND UMBILICAL CORD MATRIX IN THE DOG

Mesenchymal stem cells (MSC) are defined as multipotent stem cells that can differentiate into various cell types in vivo and in vitro under controlled conditions. These cells express specific markers detectable by analysis at the mRNA or protein level. Important sources of MSC could be fetal adnexa, such as amniotic fluid (AF), amnion (AM), and umbilical cord matrix (UCM). Canine MSC should be of use for cell-based therapies and tissue engineering improving treatment of several diseases. Moreover, the dog has been considered an attractive animal model to study human diseases. In the present study, we successfully isolated and molecularly characterised AF-MSC, AM-MSC, and UCM-MSC from dogs. Chromosomal stability and telomerase activity were also investigated. Samples were recovered after elective ovariohysterectomy in 3 bitches 25 to 40 days of gestational age. After isolation, cells were maintained in culture (Bossolasco et al. 2006 Cell Res. 16, 329–336) for different passages to perform growth and doubling time (DT) studies. Expression analyses of embryonic (Oct-4, Nanog), mesenchymal (CD44, CD184, CD29), and haematopoietic (CD34, CD45) markers were carried out by RT-PCR. Karyotype analysis was performed by Q banding. Telomerase activity was analysed by TRAPeze Telomerase Detection Kit. In all 3 cell types, the morphology of proliferating cells appeared typically fibroblast-like. In the growth study, cells isolated from AF and AM were cultured until P3, and cells isolated from UCM were maintained until P7. The population DT in AF-MSC was significantly increased (Student’s t-test: P &lt; 0.05) when comparing P1 v. P4. In AM-MSC, DT increased significantly in P1 v. P2 (P &lt; 0.001), and in UCM-MSC, DT significantly increased in P1 v. P4 (P &lt; 0.001). In AF-MSC, cell viability did not change with passages. In AM-MSC, cell viability significantly decreased (P &lt; 0.001) between P1 and P4. In UCM-MSC, cell viability remained at approximately constant levels up to P6 and significantly decreased at P7 (P &lt; 0.001). Amnion and UCM-MSC expressed Oct-4 and CD44, CD184, and CD29, whereas AF-MSC expressed only Oct-4 and CD44. Nanog, CD34, and CD45 were never found to be expressed in any cell line at any passage. In all cell lines, analysed metaphases at P4 showed normal chromosomal number and structure. Telomerase activity was observed in UCM-MSC, whereas tests on AF and AM-MSC are still on going. We first reported data on isolation, in vitro culture, and characterisation of MSC from AM and UCM in the dog. Cells expressed embryonic and MSC markers beginning at P1 and showed normal karyotype. These data indicated that canine MSC from fetal adnexa could be used to study stem cell biology and their application in therapeutic programs. Financial support was provided by Fondi di Ateneo 2009. University of Bari Aldo Moro (COD. ORBA09UDWX) (Resp. Sci. Maria Elena Dell’Aquila).

Patients with an asymptomatic short cervix (≤15 mm) have a high rate of subclinical intraamniotic inflammation: implications for patient counseling

The objective of the study was to determine the frequency and clinical significance of intraamniotic inflammation in asymptomatic women with a sonographic short cervix (SCX) in the midtrimester. This cohort study included 47 asymptomatic women (14-24 weeks) with an SCX (<or=15 mm) who underwent amniocentesis. Women with multiple gestation, cerclage, or cervical dilatation greater than 2 cm were excluded. Intraamniotic inflammation was defined as an elevated amniotic fluid (AF) matrix metalloproteinase-8 concentration (>23 ng/mL). (1) intraamniotic infection was found in 4.3% of patients; (2) among patients with a negative AF culture, the prevalence of intraamniotic inflammation was 22.2%; and (3) patients with a negative AF culture, but with intraamniotic inflammation, had a higher rate of delivery within 7 days (40% vs 5.7%; P=.016) and a shorter median diagnosis-to-delivery interval than those without intraamniotic inflammation (18 vs 42 days; P=.01). Twenty-two percent of patients with a midtrimester SCX have intraamniotic inflammation. The risk of preterm delivery within 7 days for these patients is 40%.

Amniotic fluid volume in intra-amniotic inflammation with and without culture-proven amniotic fluid infection in preterm premature rupture of membranes

Previous studies reported that the clinical significance of intra-amniotic inflammation with a negative amniotic fluid (AF) culture is similar to that of intra-amniotic inflammation with microbiologically-proven AF infection. However, the magnitude of the fetal inflammatory response in these two conditions is different as gauged by umbilical cord C-reactive protein (CRP) concentrations. We undertook this study to determine if the frequency of oligohydramnios is different in these two conditions. The amniotic fluid index (AFI) was measured in 205 patients with preterm premature rupture of membranes (PROM) (<or=35 weeks). AF was cultured for aerobic and anaerobic bacteria and genital mycoplasmas. Intra-amniotic inflammation was defined as an elevated AF matrix metalloproteinase-8 (MMP-8) concentration (>23 ng/mL). Patients were divided into three groups according to the results of AF culture and the presence or absence of intra-amniotic inflammation: 1) without intra-amniotic inflammation and a negative culture (n=109); 2) with intra-amniotic inflammation and a negative culture (n=44); and 3) a positive culture (n=52). Patients with a positive culture had a higher frequency of oligohydramnios and a lower median AFI than those with a negative culture but with intra-amniotic inflammation (P<0.01). However, there was no significant difference in the median AFI or in the frequency of oligo-hydramnios according to the presence or absence of intra-amniotic inflammation among patients with a negative culture (P>0.1). Oligohydramnios was more frequent in patients with culture-proven AF infection than in those with intra-amniotic inflammation and a negative AF culture.

532: Amniotic fluid matrix metalloproteinase-9, interleukin-6, and serum C-reactive protein in predicting histologic chorioamnionitis and intraamniotic infection

To compare the diagnostic performance of amniotic fluid (AF) matrix metalloproteinase-9 (MMP-9), interleukin-6 (IL-6), and serum C-reactive protein (CRP) in the identification of histologic chorioamnionitis and a positive AF culture.

The Frequency and Clinical Significance of Intra-Uterine Infection and Inflammation in Patients with Placenta Previa and Preterm Labor and Intact Membranes

Objective Histologic placental and/or intra-amniotic inflammation is frequently documented during ascending intra-uterine infections in patients with preterm labor and intact membranes. Placenta previa can be a clinical situation that shows the successive schema of histologic placental and intra-amniotic inflammation during the process of ascending intra-uterine infections. However, a paucity of information exists about the frequency and clinical significance of intra-uterine infections and inflammation in patients with placenta previa and preterm labor and intact membranes. The purpose of this study was to examine this issue. Study design Amniocentesis was performed on 42 patients with placenta previa and preterm labor and intact membranes (gestational age < 37 weeks). Amniotic fluid (AF) was cultured for aerobic and anaerobic bacteria and genital mycoplasmas, and AF white blood cell (WBC) count and matrix metalloproteinase-8 (MMP-8) concentrations were determined. The diagnosis of intra-amniotic inflammation was made in patients with an elevated AF MMP-8 (≥23 ng/ml). Non-parametric statistics were used for analysis. Results 1) Intra-amniotic inflammation was present in 16.7% (7/42), proven AF infection in 4.9% (2/41), and histologic chorioamnionitis in 19.0% (8/42) of patients with placenta previa and preterm labor; 2) Patients with intra-amniotic inflammation had significantly higher rates of a positive AF culture, histologic chorioamnionitis, funisitis, and a shorter interval-to-delivery than those without intra-amniotic inflammation ( p < 0.05 for each); 3) Among patients with histologic chorioamnionitis, inflammation of the choriodecidua, which was exposed to the cervical canal, existed in all cases (8/8), but inflammation of the chorionic plate existed in 63% of patients (5/8); 4) Patients with inflammation of the chorionic plate had significantly higher median AF MMP-8 concentrations and WBC counts, and higher rates of intra-amniotic inflammation than those in whom inflammation was restricted to choriodecidua ( p < 0.05 for each). Conclusions Placental inflammation was present in 19.0% and intra-amniotic inflammation was present in 16.7% of patients with placenta previa and preterm labor and intact membranes. The intra-amniotic inflammatory response was stronger when inflammation was present in the chorionic plate and choriodecidua, than when it was restricted to the choriodecidua only, which was exposed to the cervical canal in placenta previa.

Effects of Differential Glycosylation of Glycodelins on Lymphocyte Survival

Glycodelin is a human glycoprotein with four reported glycoforms, namely glycodelin-A (GdA), glycodelin-F (GdF), glycodelin-C (GdC), and glycodelin-S (GdS). These glycoforms have the same protein core and appear to differ in their N-glycosylation. The glycosylation of GdA is completely different from that of GdS. GdA inhibits proliferation and induces cell death of T cells. However, the glycosylation and immunomodulating activities of GdF and GdC are not known. This study aimed to use ultra-high sensitivity mass spectrometry to compare the glycomes of GdA, GdC, and GdF and to study the relationship between the immunological activity and glycosylation pattern among glycodelin glycoforms. Using MALDI-TOF strategies, the glycoforms were shown to contain an enormous diversity of bi-, tri-, and tetra-antennary complex-type glycans carrying Galbeta1-4GlcNAc (lacNAc) and/or GalNAcbeta1-4GlcNAc (lacdiNAc) antennae backbones with varying levels of fucose and sialic acid substitution. Interestingly, they all carried a family of Sda (NeuAcalpha2-3(GalNAcbeta1-4)Gal)-containing glycans, which were not identified in the earlier study because of less sensitive methodologies used. Among the three glycodelins, GdA is the most heavily sialylated. Virtually all the sialic acid on GdC is located on the Sda antennae. With the exception of the Sda epitope, the GdC N-glycome appears to be the asialylated counterpart of the GdA/GdF glycomes. Sialidase activity, which may be responsible for transforming GdA/GdF to GdC, was detected in cumulus cells. Both GdA and GdF inhibited the proliferation, induced cell death, and suppressed interleukin-2 secretion of Jurkat cells and peripheral blood mononuclear cells. In contrast, no immunosuppressive effect was observed for GdS and GdC.

Amniotic fluid prostaglandin F2 increases even in sterile amniotic fluid and is an independent predictor of impending delivery in preterm premature rupture of membranes

Objective. To determine whether amniotic fluid (AF) concentration of prostaglandins (PGs) increases in patients with intra-amniotic inflammation and/or proven AF infection in preterm PROM, and can predict impending delivery.Methods. AF PGF2a concentrations were determined by ELISA in 140 singleton pregnancies with preterm premature rupture of membranes (PROM) (≤35 weeks). AF was cultured for aerobic and anaerobic bacteria, and genital mycoplasmas. Intra-amniotic inflammation was defined as an elevated AF matrix metalloproteinase-8 concentration (>23 ng/ml).Results. (1) Patients with intra-amniotic inflammation and a negative AF culture had a significantly higher median AF PGF2a than those without intra-amniotic inflammation and with a negative culture (p < 0.001); (2) However, there was no difference in the median AF PGF2a between patients with intra-amniotic inflammation with a negative culture and those with culture-proven AF infection (p > 0.1); (3) Patients with an elevated AF PGF2a had a significantly shorter interval-to-delivery than those with a low AF PGF2a (≤170 pg/mL) (p < 0.001); (4) An elevated AF PGF2a (≤170 pg/mL) concentration was a significant predictor of the duration of pregnancy after adjusting for gestational age and AF inflammation/infection (p < 0.005).Conclusions. AF PGF2a (≥170 pg/mL) concentration increased in patients with intra-amniotic inflammation regardless of AF culture results. Moreover, an elevated AF PGF2a concentration was an independent predictor of impending delivery in preterm PROM.

The frequency and significance of intraamniotic inflammation in patients with cervical insufficiency

The purpose of this study was to determine the frequency and clinical significance of intraamniotic inflammation in patients with acute cervical insufficiency. Amniocentesis was performed in 52 patients with acute cervical insufficiency (cervical dilation, > or =1.5 cm) and intact membranes and without regular uterine contractions (gestational age, 17-29 weeks). Amniotic fluid (AF) was cultured for aerobic and anaerobic bacteria and genital mycoplasmas and assayed for matrix metalloproteinase-8. Intraamniotic inflammation was defined as an elevated AF matrix metalloproteinase-8 concentration (>23 ng/mL). Nonparametric statistics and survival techniques were used for analysis. The prevalence of intraamniotic inflammation was 81% (42/52); the prevalence of a positive AF culture was 8% (4/52). Intraamniotic inflammation was present in all cases with a positive AF culture. Preterm delivery within 7 days occurred in 50% of cases (19/38), and delivery before 34 weeks of gestation occurred in 84% of cases (32/38) with intraamniotic inflammation but without AF infection. Fifty-five percent of newborn infants (21/38) who were born to mothers with intraamniotic inflammation but without AF infection died immediately after birth (<1 day). The amniocentesis-to-delivery interval was shorter in patients with intraamniotic inflammation than in those without inflammation (P < .05). There were no differences in the interval-to-delivery or the rate of adverse outcome between patients with intraamniotic inflammation and a negative culture and patients with proven AF infection. Intraamniotic inflammation, regardless of AF culture result, is present in approximately 80% of patients with acute cervical insufficiency and is a risk factor for impending preterm delivery and adverse outcomes.

Immunomodulators plus antibiotics delay preterm delivery after experimental intraamniotic infection in a nonhuman primate model

The purpose of this study was to determine whether treatment with ampicillin together with dexamethasone and indomethacin delays preterm birth that is induced by intraamniotic group B Streptococcus in a nonhuman primate model. After contraction onset that was induced by group B Streptococcus (10(6) colony-forming units/mL), chronically instrumented rhesus macaques received either no treatment (controls; n = 6); ampicillin (n = 4); or ampicillin + dexamethasone + indomethacin (n = 5). Outcomes included the interval from contraction onset until delivery and concentrations of amniotic fluid inflammatory mediators. Mean interval from contraction onset until delivery was 33 +/- 8.7 hours in controls, 82 +/- 28.0 hours with ampicillin (P = .18, vs controls), and 213 +/- 50.8 hours with ampicillin + dexamethasone + indomethacin (P = .004, vs controls). Ampicillin eradicated group B Streptococcus; however, uterine activity, amniotic fluid cytokines, prostaglandins, and matrix metalloproteinase-9 remained elevated. Ampicillin + dexamethasone + indomethacin suppressed interleukin-1beta, tumor necrosis factor-alpha, and prostaglandins E2 and F2alpha but did not alter matrix metalloproteinase expression or chorioamnionitis. The combination of ampicillin + dexamethasone + indomethacin suppressed inflammation and significantly prolonged gestation.

The intensity of the fetal inflammatory response in intraamniotic inflammation with and without microbial invasion of the amniotic cavity

Intraamniotic inflammation is a risk factor for adverse pregnancy and neonatal outcome, regardless of the presence or absence of a positive amniotic fluid (AF) culture. The purpose of this study was to determine whether the intensity of a fetal inflammatory response (FIR) differs between cases of intraamniotic inflammation with microbiologically proven infection and cases with negative AF cultures. The FIR was examined in 89 cases of women with preterm premature rupture of membranes who delivered singleton preterm newborn infants within 48 hours of amniocentesis. AF was cultured for aerobic and anaerobic bacteria and for genital mycoplasmas. AF white blood cell (WBC) count and matrix metalloproteinase-8 (MMP-8) determinations were performed to assess the presence of intraamniotic inflammation. Intraamniotic inflammation was defined as an elevated AF MMP-8 concentration (>23 ng/mL). The intensity of the FIR was determined by the umbilical cord plasma concentrations of C-reactive protein (CRP). Patients were divided into 3 groups according to the presence or absence of intraamniotic inflammation and AF culture results: group 1, without intraamniotic inflammation and with a negative AF culture (n = 28); group 2, with intraamniotic inflammation and with a negative AF culture (n = 26); group 3, with a positive AF culture (n = 35). Neonates who were born to mothers with intraamniotic inflammation and negative AF cultures had a significantly higher median umbilical cord plasma CRP concentration than did those without intraamniotic inflammation and a negative AF culture (P < .005) but a significantly lower median cord plasma CRP concentration than did those with proven AF infection (P < .05). Patients with intraamniotic inflammation and a negative AF culture had significantly higher median AF MMP-8 concentrations and WBC count than did those without intraamniotic inflammation and a negative AF culture (P < .001). However, there was no significant difference in the median AF MMP-8 and WBC count between patients with intraamniotic inflammation and a negative AF culture and those with proven AF infection (MMP-8, P > .1, and WBC, P = .09). Intraamniotic inflammation without documented AF infection is a risk factor for a systemic FIR. However, the magnitude of the FIR in those cases was lower than in those with documented AF infection.

C-reactive protein concentration in vaginal fluid as a marker for intra-amniotic inflammation/infection in preterm premature rupture of membranes

Objective. The purpose of this study was to determine whether C-reactive protein (CRP) concentrations in vaginal fluid can identify patients with intra-amniotic inflammation/infection (IAI) and predict adverse outcome in preterm premature rupture of membranes (PROM).Methods. The study population consisted of 121 singleton pregnant women with preterm PROM (⩽36 weeks of gestation) who had an amniocentesis and vaginal fluid collection. A Dacron polyester-tipped applicator was soaked with vaginal fluid for 10 seconds and diluted with 1 mL buffer solution. Amniotic fluid was cultured for aerobic and anaerobic bacteria, as well as mycoplasmas. Vaginal fluid CRP and amniotic fluid matrix metalloproteinase-8 (MMP-8) were determined by specific immunoassays. IAI was defined as an amniotic fluid MMP-8 concentration >23 ng/mL and/or a positive amniotic fluid culture. Nonparametric tests and survival techniques were used for statistical analysis.Results. Patients with IAI had a significantly higher median vaginal fluid CRP concentration than those without IAI (median (range), 7.8 (0.1–1310.1) ng/mL vs. 1.0 (0.1–319.4) ng/mL, p < 0.005). The median amniotic fluid white blood cell (WBC) count was significantly higher in patients with a vaginal fluid CRP concentration of >10 ng/mL than in those with a lower concentration (median (range), 82.5 (0–8640) cells/mm3 vs. 2 (0–>1000) cells/mm3, p < 0.001). Patients with vaginal fluid CRP concentration of >10 ng/mL had a significantly shorter sampling-to-delivery interval and higher rates of preterm delivery within five days, funisitis, and histologic chorioamnionitis than did those with a vaginal fluid CRP concentration below this cut-off. A vaginal fluid CRP cut-off of 10 ng/mL had a specificity of 89% and a sensitivity of 45% in the identification of IAI.Conclusion. An elevated CRP concentration in vaginal fluid collected by polyester-tipped applicator is a risk factor for intra-amniotic inflammation/infection and impending preterm delivery in preterm PROM.