Tomomi Aoyama

Elevation of Transforming Growth Factor-β1 Is Associated with Recurrent Miscarriage

To investigate the significance of transforming growth factor-β1 (TGF-β1) in reproduction we have compared plasma levels in normal pregnant women and patients suffering miscarriages. We examined 188 normal pregnant women and 12 pregnant women with miscarriages. Eight women with severe recurrent miscarriages (mean ± SD of previous number of miscarriages; 10.4 ± 2.4 times) were also examined before conception; 34 nonpregnant women served as controls. Plasma TGF-β1 level increased with the gestational week and returned within the normal range 1 month after delivery. The levels among pregnant women with miscarriages (mean ± SD; 2.44 ± 0.83 ng/ml) were significantly higher than those of pregnant controls (1.74 ± 0.95 ng/ml) of matched gestational weeks; levels among nonpregnant women with severe recurrent miscarriages were extremely elevated (4.1 ± 3.04 ng/ml) compared to the control value (1.34 ± 0.59 ng/ml). These data suggest that TGF-β1 may be necessary to maintain pregnancy but also may be a risk factor for recurrent miscarriages.

Low Serum M-CSF Levels Are Associated with Unexplained Recurrent Abortion

PROBLEM: The purpose of this study was to determine whether the serum macrophage‐colony stimulating factor (M‐CSF) level is associated with early pregnancy loss in unexplained recurrent spontaneous abortion (RSA) patients.

Are serum progesterone levels predictive of recurrent miscarriage in future pregnancies

OBJECTIVE To examine the predictive value of midluteal serum P as a marker of a luteal phase defect (LPD) in future pregnancies of recurrent aborters. DESIGN Prospective analysis. SETTING Nagoya City University Hospital. PATIENT(S) One hundred ninety-seven women with a history of two consecutive first trimester abortions, none of whom had any other medical problems or an identifiable cause of recurrent miscarriages, such as uterine anomalies or evidence of antiphospholipid antibodies. None of the study subjects received any medication for miscarriage or infertility. MAIN OUTCOME MEASURE(S) A midluteal phase single serum P level < 10 ng/mL was used as the criterion for a potential LPD: those whose subsequent pregnancy was successful and those in which failure was the end result. RESULT(S) Of the 197 patients, 46 (23.4%) demonstrated LPD without other endocrine abnormalities and 38 (19.3%) recurrent aborters suffered another abortion, with figures for LPD-negative and LPD-positive patients of 20.5% (31/151) and 15.2% (7/46), respectively. There was no statistically significant difference between the two groups. CONCLUSION(S) Progesterone, E2, and the P/E2 ratio may not predict future pregnancy loss in recurrent aborters.

Prevalence of autoantibodies in patients with recurrent miscarriages.

PROBLEM: It is well known that the prevalence of several autoantibodies is higher in patients with recurrent miscarriages than in normal women. However, links between individual autoantibodies are unclear. The present study focuses on the possible association between β2‐glycoprotein I (β2‐GPI)‐dependant anticardiolipin antibody (aCL), lupus anticoagulant (LA), and antinuclear antibody (ANA) in patients with recurrent miscarriages.

Recurrent abortion and moderate or strong antiphospholipid antibody production

Objective: To investigate the treatment outcome for women suffering recurrent miscarriages associated with strong or moderate antiphospholipid antibody (aPL) production. Methods: Sixty‐seven pregnancies in 61 women demonstrating at least one kind of aPL with a history of recurrent miscarriages were treated with: (1) aspirin (ASA) alone; (2) prednisolone (PSL) and ASA; and (3) PSL, ASA, heparin and/or immunoglobulin (IgG). For comparison purposes the aPL‐positive patients were divided into two groups, strongly and moderately‐positive. IgG and IgM antibodies against PE and five negatively‐charged phospholipids were measured by ELISA between 1987 and 1993. β2‐glycoprotein I (β2GPI) dependent anticardiolipin antibodies were measured by ELISA since 1993. Lupus anticoagulant was measured by a diluted aPTT method since 1993. Results: Out of a total of 16 (50%) patients strongly‐positive for aPL and 47 out of 51 (92.2%) moderately‐positive demonstrated a successful outcome. The live birth rate moderate group was significantly higher than in the strongly‐positive cases (P<0.0005). In the cases exhibiting moderate aPL production, 28 out of 30 (93.3%) receiving PSL and ASA and 14 out of 15 (93.3%) treated with ASA alone successfully gave birth. None of the 14 given ASA alone suffered preterm delivery or IUGR. In contrast 12 (36.4%) and 6 (18.2%) of the 33 patients treated with the PSL combination therapy suffered from preterm delivery and IUGR, respectively. Conclusions: The live birth rate in patients strongly positive for aPL is lower than that in patients with moderate aPL production even if treatment is performed during pregnancy. However, ASA is useful to treat cases with moderate aPL so that distinction of the two groups is warranted.

Clinical trial of immunostimulation with a biological response modifier in unexplained recurrent spontaneous abortion patients

We determined clinically whether a killed streptococcal preparation (KSP), a biological response modifier, is as effective as paternal lymphocyte immunotherapy for unexplained recurrent pregnancy abortion (RSA) therapy. The success rate of adverse pregnancy in the study group of 23 RSA cases, who were administered low doses of KSP before and during early pregnancy, was statistically compared with that in a control group of 205 women who received paternal lymphocyte immunotherapy. The association of natural killer (NK) cell activity in the peripheral blood with pregnancy outcome was also assessed. The success rate in the study group was 73.9% (17/23), compared to 75.1% (154/ 205) observed for the controls (not significant). Most of the successful cases exhibited low levels of NK cell activity in the peripheral blood. Immunotherapy with low doses of KSP is as effective as that with paternal lymphocytes, providing a simple and safe alternative therapy for unexplained RSA. Suppression of NK cell activity by some immunoregulatory mechanism was also found to have potential benefit in terms of a successful pregnancy outcome.

Involvement of μ‐Calpain in Human Sperm Capacitation for Fertilization

PROBLEM: The distribution and physiological role of calpains in human sperm were investigated.
 METHOD OF STUDY: Semen collected manually from healthy donors was liquefied then centrifuged by percoll gradient centrifugation. After exposure to different concentrations of Ca2+ ionophore A23187, the samples were used for immunostaining sodium dodecyl sulfate polyacrylamide gel electrophoresis, and western blot analysis. It was speculated from the results of the study using calpain‐specific inhibitors that calpain contributes to the sperm motility and acrosome reaction.
 RESULTS: With the anti‐pro μ‐calpain antibody, sperm were immunostained, whereas all were negative for anti‐pro m‐calpain antibody binding. Stained sperm were classified into four types according to the staining pattern: acrosome type, equatorial segment type, whole head type, and neck and tail segments type. Western blot analysis of sperm homogenate revealed a single 80‐kDa band using the anti‐pro μ‐calpain antibody, its dose‐dependent reduction with Ca2+ ionophore A23187 suggesting activation by this treatment. In the presence of membrane permeable calpain inhibitors, sperm motility and acrosome reaction were significantly suppressed.
 CONCLUSION: These results indicate that μ‐calpain may play pivotal roles in the process of fertilization of human sperm.

Activated partial thromboplastin time is a predictive parameter for further miscarriages in cases of recurrent fetal loss

OBJECTIVE To determine whether clinically routine clotting tests such as activated partial thromboplastin time (aPTT), prothrombin time (PT), or fibrinogen can be used to predict further miscarriages. DESIGN Prospective study. SETTING Nagoya City University Hospital, Nagoya, Japan. PATIENT(S) A total of 261 patients with a history of two consecutive first-trimester spontaneous abortions who had no antiphospholipid antibodies or other autoimmune diseases and no anatomic anomalies were examined for aPTT, PT, and fibrinogen before becoming pregnant again. INTERVENTION(S) Blood tests were performed before pregnancy. Patients then were followed up during subsequent pregnancy and their outcomes were compared with their previous blood test results. MAIN OUTCOME MEASURE(S) Activated partial thromboplastin time, PT, and fibrinogen were measured by coagulation time methods. RESULT(S) Fifty-eight of 261 patients (22.2%) had a subsequent miscarriage. Mean (+/-SD) values for preconception aPTT in individuals whose subsequent pregnancies ended in success and failure were 88.2%+/-23.4% and 99.3%+/-26.4%, respectively. The difference was statistically significant. Respective values were 106.8%+/-22.8% and 106.3%+/-21.4% for PT and 245+/-61.1 mg/dL and 259.1+/-57 mg/dL for fibrinogen. These findings were not significantly different. CONCLUSION(S) A shortened aPTT before conception is associated with further miscarriages in patients with a history of recurrent spontaneous abortions who have no antiphospholipid antibodies.

Clinical Significance of β2 Glycoprotein I Dependent Anticardiolipin Antibody, Lupus Anticoagulant and Antinuclear Antibodies in Patients with Recurrent Miscarriages

Antiphospholipid antibodies (aPL) are common as a predictive variable for autoimmunity related miscarriages. However, it is controversial whether antinuclear antibody (ANA)-positive patients should be treated with any medication, because it is unclear whether ANAs are in fact pathogenic. The present study focuses on the possible association betweenβ2 glycoprotein IQ32GPI) dependent anticardiolipin antibody (aCL), lupus anticoagulant (LA) and ANA and the clinical significance of ANA in patients with recurrent miscarriages. We have studied 301 patients with a history of two or more miscarriages. β2GPI dependent aCL were measured by the ELISA method. LA was detected by the dilute aPTT method. ANAs were measured by indirect immunofluorescence on Hep-2 cell slides. We also studied the subsequent pregnancy outcome in 225 patients with a history of two consecutive first-trimester abortions, focusing on ANA. None of the 225 patients had aPL and received any medication or immunotherapy. The prevalence of β2GPI dependent aCL, LA and ANA were 3.3% (10 of 301), 10.0% (30 of 301) and 25.2% (76 of 301), respectively. Four of 301 patients had all antibodies. Regarding ANA in 225 patients with a history of two miscarriages, 39 of the 225 (17.3%) patients had ANAs, as compared with 33 of 740 (4.5%) control women. The prevalence of ANA in patients was significantly higher than that of controls (p < 0.001, Odds Ratio = 4.49). Forty-nine of the 225 (21.8%) recurrent aborters suffered a further abortion, the figures for ANA-negative and ANA-positive patients being 23.1% (43 of 186) and 15.4% (6 of 39) respectively (no significant difference).

Clinical Significance of Anti-GM3 Antibodies in Recurrent Pregnancy Loss With Elevated Level of Antiphospholipid Antibodies

PROBLEM: The ganglioside‐GM3 neutralizes the reactivity of antiphospholipid antibodies (APLs) to phospholipids in vitro. The question of whether anti‐GM3 antibodies might exert influence in APLs‐positive recurrent pregnancy loss patients who are undergoing prednisolone and aspirin (PSL/ASA) treatment was investigated.