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Genetic prothrombotic risk factors in women with unexplained pregnancy loss.

INTRODUCTION: Inherited thrombophilia has been associated with unexplained recurrent pregnancy loss (RPL) and stillbirth. This thrombotic tendency can manifest as thrombotic lesions in the placenta, and may lead to abortion and stillbirth. The aim of our case-control study was to investigate the prevalence of FVL and FII G20210A in women with adverse pregnancy outcome, compared to the prevalence of the same mutations in our health control group. MATERIALS AND METHODS: 102 consecutive women with unexplained pregnancy loss (55 with history of RPL, and 47 with history of stillbirth) were studied for hereditary thrombophilia. The health control group consisted of 217 healthy women from the general population. RESULTS AND CONCLUSIONS: Of the 55 women with recurrent abortions, we found the same prevalence for the FVL and the FII G20210A(9.1%, 5 pts). (p=NS compared to control group). Of the 47 women with stillbirth, 11 (23.4%) had the FVL and 9 (19.1%) had the FII G20210A(p<0.0005 for both mutations). In our experience the prevalence of FVL and the FII G20210Amutations was significantly higher in women with unexplained stillbirth, instead the prevalence of genetic thrombophilia was high but not statistically significant in women with recurrent pregnancy loss.

Abortion, Habitual↗

Purification and characterization of a lectin from seeds of Vatairea macrocarpa Duke.

A lectin from Vatairea macrocarpa Duke seeds (VML) was isolated using affinity chromatography on a guar gum column. The lectin, a glycoprotein without erythrocyte specificity, displays specificity to galactose and some derivatives. On SDS-polyacrylamide gels, V. macrocarpa seed lectin is composed of two major high-Mr bands of 34 and 32 kDa and two minor low-Mr bands of 22 and 13 kDa. N-Terminal sequencing showed that the 34, 32, and 13 kDa products possess identical N-terminal sequence, which display best similarity with the N-terminal portion of Robinia pseudoacacia lectins (RPL). On the other hand, the N-terminal sequence of the 22 kDa band can be aligned with an internal sequence of RPL starting at residue 149 of the cDNA-derived sequence. These data indicate that, like other leguminous lectins, VML is made up of a mixture of one-chain 30-35 kDa glycoforms and of 22 and 13 kDa endogenous C- and N-terminal fragments. Size-exclusion chromatography indicated that, at neutral pH, VML is predominantly a dimeric (70 kDa) protein, although tetramers (115 kDa) and larger aggregates (300 kDa) were also present.

Amino Acids↗

Field studies on imposex and organotin accumulation in the rock shell, Thais clavigera, from the Seto Inland Sea and the Sanriku region, Japan.

Field studies on imposex and organotin (butyltin and phenyltin) contamination in the Japanese molluscs (the rock shell, Thais clavigera, and the Pacific oyster, Crassostrea gigas) were conducted along the coast of both the Seto Inland Sea and the Sanriku region, Japan, during February-March 1996. The percentage occurrence of imposex in the rock shell, Thais clavigera, was still 100% at all sites surveyed (22 and seven sites in the Seto Inland Sea and the Sanriku regions, respectively). Observed values of the Relative Penis Length (RPL) index (%), defined as the ratio of mean penis length in females to that in males, exceeded 50 at many sites. Sterile individuals whose oviducts were blocked by vas deferens formation were also extensively observed. Tissue concentrations of organotin compounds in the rock shell and the Pacific oyster were higher than those from other areas of Japan. The degree of imposex (RPL index) was correlated with the sum of TBT and TPT concentrations in tissue of the rock shell. Less recovery from imposex in the rock shell and less amelioration of TBT pollution were observed, suggesting that organotin use in antifouling paints has still continued for vessels larger than 25 m in length and that it has influenced imposex symptoms in the rock shell.

Animals↗

Cytokine promoter gene polymorphisms and idiopathic recurrent pregnancy loss.

Approximately one in 300 women experience recurrent pregnancy loss (RPL), the aetiology of which is unknown in at least 40% of cases. Previously, some studies have shown increased production of pro-inflammatory cytokines (tumour necrosis factor-alpha and interferon-gamma) and reduced production of anti-inflammatory cytokines (interleukin-10) by circulating blood lymphocytes isolated from these patients when compared with controls. The reasons for this are unclear. The production of these cytokines are partly under genetic control. This study investigated whether polymorphisms in these three cytokine genes known to be associated with either high or low production, are associated with idiopathic RPL. No association was found. It may be that genetic factors are not a major determinant of cytokine production during pregnancy, or alternatively it may be that the observed differences in cytokine production by peripheral lymphocytes do not accurately indicate what is occurring at the local maternofoetal interface during successful and abortive pregnancies.

Abortion, Habitual↗

Cytotoxic T cell induction with ratchet peptide libraries.

Immunization with synthetic peptides are used to induce cytotoxic T cell (CTL) responses in vivo. However, CTL peptide vaccines require the use of multiple peptides to overcome genetic diversity associated with MHC restriction, and prior epitope identification from the chosen protein template. We describe here a method whereby all nonamer sequences from a longer template can be synthesized simultaneously in a ratchet peptide library (RPL) covering all potential epitopes within a protein. We synthesized an RPL based on a template sequence from the Plasmodium berghei circumsporozoite (CS) protein (CSRPL). Using a lipopeptide formulation we immunized mice i.p. with the CSRPL and elicited CS specific CTL, which recognized the CS252-260 H-2Kd restricted CTL epitope.

Amino Acid Sequence↗

Imposex in the commercial snail Bolinus brandaris in the northwestern Mediterranean.

A survey of the degree of imposex in populations of the commercial muricid Bolinus brandaris (L.) was carried out at six locations along the Catalan Coast (NW Mediterranean). Imposex was found to be a widespread phenomenon; the occurrence of a penis was observed in all females studied with the exception of those from St. Carles (South of Catalunya) where only 37% were affected. Other indices such as Relative Penis Length (RPL) and Vas Deferens Sequence (VDS) were applied; RPL varied from 4.5% in St. Carles--the least affected area--to 29.8% in the vicinity of Barcelona; similar results were obtained for VDS. Tissue organotin residues ranged from 6 to 140 ng g(-1) w.w. as Sn. Despite legislation restricting the use of tributyltin (TBT) in antifouling paints, this paper evidences the occurrence of organotin pollution far from the source, with levels of both TBT and triphenyltin (TPhT) high enough to cause environmental concern.

Journal Article↗

Interaction of RNA with phage display selected peptides analyzed by capillary electrophoresis mobility shift assay.

A sensitive capillary electrophoresis mobility shift assay (CEMSA) to analyze RNA/peptide interactions has been developed. Capillary electrophoresis (CE) has been adapted for investigating the interaction between variously methylated 17-nt analogs of the yeast tRNAPhe anticodon stem and loop domain (ASL(Phe)) and 15-amino-acid peptides selected from a random phage display library (RPL). A peptide-concentration-dependent formation of RNA/peptide complex was clearly visible during CEMSA. In the presence of peptide, the UV-monitored CE peak for ASLPhe with three of the five naturally occurring modifications (2'-O-methylcytidine (Cm32), 2'-O-methylguanine (Gm34) and 5-methylcytidine (m5C40) shifted from 18.16 to 20.90 min. The mobility shift was observed only for methylated RNA. The negative effects of diffusion, electroosmotic flow and adhesion of molecules to the capillary internal wall were suppressed by using a buffer containing a sieving polymer and a polyacrylamide-coated capillary. Under these conditions, well-shaped peaks and resolution of RNA free and bound to peptide were achieved. Peptide tF2, the most populated ligand in the RPL, specifically bound triply methylated ASLPhe in a methylated nucleoside-dependent manner. CE was found to be an efficient and sensitive method for the qualitative analysis of RNA-peptide interaction and should be generally applicable to the study of RNA-peptide (protein) interactions.

Amino Acid Sequence↗

Genetics of recurrent pregnancy loss.

Recurrent pregnancy loss (RPL) is a devastating reproductive problem affecting approximately 5% of couples trying to conceive. Genetic factors appear to be highly associated with reproductive loss. In this article, genetic factors are reviewed in terms of random numerical chromosome errors in miscarriage specimens and carriers of structural chromosome rearrangements that may result in unbalanced chromosome errors in pregnancies. Recently, research has generated interest in genetic markers for recurrent loss such as skewed X-chromosome inactivation and human leukocyte antigen-G polymorphisms. Assisted reproductive technologies (specifically, preimplantation genetic diagnosis) have been offered to couples with recurrent pregnancy loss; however, more data need to be evaluated before routine use can be advocated. Management of genetic factors in RPL should include therapy based on the highest level of evidence, genetic counseling, and close monitoring of subsequent pregnancies.

Abortion, Habitual↗

Comparison of gene expression at the feto-maternal interface between normal and recurrent pregnancy loss patients.

Normal pregnancy requires a series of immunological, metabolic, vascular and endocrine regulating processes. However, the specific genes and proteins involved in these processes are not well defined. Aberration of these processes may lead to problems in pregnancy. One of these problems may be recurrent pregnancy loss (RPL). Little information is available on the level of expression of genes that may play a role in normal pregnancy. Therefore, this study determined whether different levels of gene expression at the feto-maternal interface could be associated with factors for RPL. The expression patterns of genes isolated from subtractive hybridization analysis performed with chorionic villi from normal and abnormal pregnancies were investigated. Eight genes classified into groups, including immunosuppression-related, embryo attachment-related and angiogenesis-related, were isolated.

Abortion, Habitual↗

Assessment of periodontal healing by seeding of fibroblast-like cells derived from regenerated periodontal ligament in artificial furcation defects in a dog: a pilot study.

The regeneration of periodontal supporting tissues lost as a result of disease could be accomplished by repopulating the exposed root surfaces with cells originating from periodontal ligament. Thus, we aimed to assess the seeding of cells derived from regenerated periodontal ligament (RPL) to promote the regeneration in artificial furcation defects of a dog. The fibroblast-like cells were obtained by incubating the explants of RPL tissue taken under a teflon (E-PTFE) membrane. Class II furcation defects were induced on the second and fourth mandibular premolars. Control defects were also included on the contralateral side. A suspension of the fourth passage cells (2 x 10(5) cells) in 0.5 mL of autologous blood coagulum was placed over each furcation area. The healing was histomorphometrically evaluated at the 42nd day postoperatively and expressed as percentage. The healing by new connective tissue attachment with cementum formation was found 75% in the cell-seeding defects whereas, it was 71% in controls. Bone formation was found to fill 51% of furcation defects; however, it was 35% of the defects in the control sites. In this pilot study, we suggested that regeneration of furcation defects by cell-seeding technique may be useful, but further studies are needed to determine the outcome of the procedure.

Alveolar Process↗

Types of pregnancy loss in recurrent miscarriage: implications for research and clinical practice.

BACKGROUND: In recurrent pregnancy loss, there is much debate about cause and association, as exact pathophysiological mechanisms have not been elucidated. The aim of this study was to assess whether recurrent pregnancy loss (RPL) patterns differ according to causal/associated conditions, suggesting differing disease processes. METHODS: Following investigation, 427 women with RPL were classified into the following 'diagnostic' groups: idiopathic, oligomenorrhoea, antiphospholipid syndrome (APS) and 'possible' APS. A total of 323 subsequently conceived, and underwent serial ultrasonography in early pregnancy; of these, 87 (26.9%) miscarried, and the types of pregnancy loss for the four diagnostic groups were allocated to either embryo loss (fetal heart never seen) and fetal loss (fetal heart seen prior to pregnancy loss). RESULTS: Overall, there were 75 embryonic losses and 12 fetal losses. The fetal loss rates in each group were similar: idiopathic 5.1%, oligomenorrhoea 3.4%, 'possible' APS 4.9% and APS 4.8%. CONCLUSIONS: Serial ultrasound helps to discriminate type of pregnancy loss and demonstrates that embryo loss is more common than fetal loss. More importantly, pregnancy loss patterns do not seem to differ between diagnostic groups in a treated population. The fetal loss rate in each of the diagnostic groups is similar to that in other reported populations.

Abortion, Habitual↗

Homing and in situ differentiation of resident pulmonary lymphocytes.

At birth, T lymphocytes which colonize the lung are mainly of the gamma delta subset, while alpha beta T cells predominate in the spleen. Thus, the lung is a preferred site for the homing of gamma delta T cells in the perinatal period. However, after birth, the pattern of V gamma gene usage among resident pulmonary lymphocytes (RPL) changes with age, from a predominance of V gamma 6 at birth to a predominance of V gamma 4 in older mice. The generation of the V gamma 6 fraction appears to be thymus dependent, since in athymic nude mice, the V gamma 6 population present at birth is replaced by V gamma 4 T cells. In the postnatal period, both RAG-1 and RAG-2 genes are expressed at high levels in the RPL population. TCR bearing cells are among those that express RAG genes, indicating that maturation of T cells takes place in this organ. In addition, transfer experiments reveal that lymphoid precursors are present in the lung. The stage of differentiation of these precursors will be characterized in future studies. The data presented here indicate that pulmonary T lymphocytes are derived from both migrants of thymic origin and from precursors which have undergone differentiation and selection in the lung. The population that is generated in situ and that has not been selected in the thymus may include cells that are typical for the pulmonary environment.

Actins↗

Aberrant gene expression associated with recurrent pregnancy loss.

Recent studies indicate that a number of factors including chromosomal abnormalities, immunological feto-maternal rejection, hormonal irregulation and anatomical factors are involved in provoking recurrent pregnancy loss (RPL). This indicates that normal cellular regulation of these factors is required for maintaining normal pregnancy. In addition, it is expected that biological processes for maintaining normal pregnancy require a series of differential gene expression. As expected, our previous investigations revealed that there are >/=30 genes showing different levels of expression between normal and RPL patients. In addition, other research groups have also identified a number of genes that are expressed aberrantly in pregnancy failure. In this review, recent study on aberrant expression levels of genes, which are grouped as immunity-related, angiogenesis-related, apoptosis-related and other groups of genes, will be discussed.

Abortion, Habitual↗

Impaired expression of endometrial differentiation markers and complement regulatory proteins in patients with recurrent pregnancy loss associated with antiphospholipid syndrome.

Antiphospholipid syndrome (APS), characterized by circulating antiphospholipid (aPL) antibodies, is a major cause of early pregnancy failure and placental insufficiency. In this study, we examined whether impaired endometrial differentiation before conception contributes to the high incidence of pregnancy complications in APS. Timed secretory endometrial biopsies were obtained from a cohort of women with recurrent pregnancy loss (RPL). Real-time quantitative (RTQ)-PCR was used to determine the expression levels of transcripts that encode for decidual markers, proinflammatory cytokines and complement regulatory proteins. Expression of decidual markers such as prolactin (PRL), tissue factor (TF) and signal transducer and activator of transcription 5 (Stat5), but not insulin-like growth factor-binding protein 1 (IGFBP-1), was significantly lower in samples obtained from aPL(+) patients (n = 24) when compared with aPL(-) group (n = 58) (P < 0.05). The abundance of transcripts encoding for interferon gamma (IFNgamma), tumour necrosis factor alpha (TNFalpha) or Stat1 did not differ significantly between both groups (P >/= 0.05). However, analysis of transcripts that encode for complement regulatory proteins showed a marked decrease in decay-accelerating factor (DAF/CD55) levels in aPL(+) patients (P = 0.005), which was mimicked at protein level as demonstrated by immunohistochemistry. In summary, patients with RPL have distinct endometrial gene expression profiles depending on the presence or absence of circulating aPL antibodies. In APS, impaired endometrial differentiation and lower DAF/CD55 expression before conception may compromise implantation and predispose to complement-mediated pregnancy failure.

Abortion, Habitual↗

Temporal concentrations of rat placental lactogen in the peripheral circulation.

Placental lactogen (PL) is an abundant placental protein identified in several species, but to date little is known of mode or control of PL secretion. In the present study we sought to define the minute-to-minute variation in rat PL (rPL) secretion in late pregnancy by repeated blood sampling over a 3-h period. The results indicate that rPL may be secreted in an episodic pulsatile manner similar to the pulsatile secretion of pituitary prolactin (Prl) and growth hormone (GH). This evidence suggests a highly regulated dynamic control of secretion of a placental hormone.

Animals↗

Hormonal regulation of copper-zinc superoxide dismutase and manganese superoxide dismutase messenger ribonucleic acid in the rat corpus luteum: induction by prolactin and placental lactogens.

The corpus luteum expresses two enzymes that scavenge superoxide radicals and protect the cells from their toxic activities: cytosolic copper, zinc-superoxide dismutase (Cu,Zn-SOD) and mitochondrial manganese-SOD (Mn-SOD). The present study was undertaken to investigate whether the mRNA expression of each of these enzymes is regulated by luteotropic hormones. Cu,Zn-SOD and Mn-SOD mRNA levels were determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). We first examined the effects of prolactin (PRL) on Cu,Zn-SOD and Mn-SOD mRNA expression in the corpus luteum. Hypophysectomy of Day 3 pregnant rats caused a sharp decline in both Cu,Zn-SOD and Mn-SOD mRNA levels, which was completely reversed by PRL administration. To further examine the effects of PRL and rat placental lactogen (rPL) on the expression of these enzymes, either primary luteinized granulosa cells or temperature-sensitive simian virus-40 transformed luteal cells (GG-CL) were cultured with different doses of PRL or rPL. These hormones induced a remarkable increase in Cu,Zn-SOD and Mn-SOD mRNA levels in both primary luteinized granulosa cells and GG-CL cells. Interestingly, whereas PRL up-regulated the expression of the SOD in luteal cells, other luteotropic hormones such as estradiol and dexamethasone inhibited both SOD mRNA expression while progesterone had no effect. In conclusion, PRL and PRL-like hormones induce a protective ability against toxic oxygen radicals by stimulating the expression of SODs, a phenomenon that may play an important role in maintaining luteal cell integrity and steroidogenic capacity.

Animals↗

Antiphosphatidylethanolamine antibodies in recurrent early pregnancy loss and mid-to-late pregnancy loss.

AIM: Associations have been reported between antiphospholipid antibodies (aPL), mainly anticardiolipin antibodies (aCL) and/or the lupus anticoagulant, and recurrent pregnancy losses (RPL). However, relatively few studies describing antiphosphatidylethanolamine antibodies (aPE) have been reported. We describe the prevalence of aPL to both cardiolipin and phosphatidylethanolamine in patients with RPL. METHODS: Patients with recurrent early pregnancy losses (n = 145) and mid-to-late pregnancy loss(es) (n = 26) were screened for aPE and aCL. RESULTS: In patients with recurrent early pregnancy losses, prevalence of immunoglobulin G (IgG) aPE (17.9%, P = 0.001) and immunoglobulin M (IgM) aPE (12.4%, P = 0.01) was significantly higher than in the control group. In patients with mid-to-late pregnancy loss(es), prevalence of IgM aPE (19.2%, P = 0.008) and IgG aCL (23.1%, P = 0.02) was significantly higher than in the control group. CONCLUSION: Our data suggest that aPE may be a risk factor in patients with mid-to-late pregnancy loss(es) as well as recurrent early pregnancy losses.

Abortion, Habitual↗

Laboratory evaluation of women experiencing reproductive failure.

Reproductive life table analysis indicates that the majority of reproductive failures result from post fertilization failures, whether before or after implantation. It is important to have a set of tests to clarify the diagnosis of the reproductive failure so that appropriate therapy can be instituted. To determine the frequency of abnormal immunologic tests among women experiencing reproductive failure, 108 patients were evaluated for the presence of antiphospholipid antibodies (APA); lupus anticoagulant (LA); thyroid-thyroglobulin and microsomal antibodies (TGT); embryotoxic factor (ETA); and systemic CD56+/CD16- cells. The frequency of abnormal results obtained from testing for APA, LA, TGT, ETA, and CD56+/CD16- cells among 108 patients with diagnoses of recurrent pregnancy loss (RPL)(n = 45), unexplained infertility (n = 45) including IVF failure (n = 10), endometriosis (n = 10), premature ovarian failure (n = 5), and polycystic ovaries (n = 3) were compared with 15 normal controls. Seventy of one hundred eight (65%) women experiencing reproductive failure had at least one positive test, compared to 1 of 15 (7%) controls (P = 0.0001). Presence of phospholipid antibodies was the most frequently abnormal result followed by elevated CD56+/CD 16 cells. The prevalence of a particular abnormal test varied among the diagnoses. The most frequent abnormal test among women with RPL was an increased percentage of CD56+/CD16- cells (40%), followed by APAs (29%), TGT (9%), and ETA (7%). The most frequent abnormal result among women with unexplained infertility was the presence of APAs (42%), followed by CD56+/CD16- cells (16%), ETA (16%), and TGT (9%). APA, CD56+/CD16- cells, ETA, and TGT are useful tools to assist in the diagnosis of reproductive failure.

Abortion, Habitual↗