bluuebunny/crossref_metadata_2025 huggingface dataset:
Abnormal TGFβ Levels in the Amniotic Fluid of Down Syndrome Pregnancies
Authors: BROMAGE STEPHEN J. , LANG ALLISON K. , ATKINSON IAN , SEARLE ROGER F.
Published: December 2003

PROBLEM: The status of cytokines in amniotic fluid (AF) from chromosomally abnormal pregnancy is largely undefined. TGFβ plays a key role in fetal growth and differentiation and is responsible for the immunoregulatory activity of AF in normal pregnancy, but its status in Down syndrome (DS) pregnancies is unknown. In addition we investigated the IL‐2 status of AF from DS pregnancies.  METHOD OF STUDY: Midtrimester AF from chromosomally normal (n=25)
and abnormal pregnancies with DS (n=15) were assayed for bioactive
and latent TGFβ levels using the mink lung epithelial cell growth
inhibition bioassay and for IL‐2 activity by the CTLL‐2 cell
proliferation bioassay and by ELISA. 
RESULTS: Levels of bioactive TGFβ (mean 4.6±0.6 U/mL) were
significantly increased in DS AF compared with the normal samples (mean
2.8±0.3 U/mL, P<0.003) but latent TGFβ levels did
not differ between DS and normal groups. In addition DS AFs showed
reduced IL‐2 like bioactivity compared with normal samples
(P=0.0006) but IL‐2 immunoactivity was undetectable in DS and normal AFs by ELISA.  CONCLUSIONS: DS AFs display increased TGFβ activity and lacked IL‐2 immunoactivity. The reduced ability of DS AFs to stimulate CTLL‐2 cell proliferation is unrelated to the IL‐2 status of AF. Altered TGFβ levels may prove useful as an additional biochemical index for the detection of DS pregnancies.
Early Pregnancy Factor Activity in Serum of Patients with Surgical Abortion
Authors: CHENG SHU‐JUAN , MA AI‐YING , QIAO CHANG‐XIN , ZHENG ZHEN‐QUN
Published: December 2003

PROBLEM: The presence of early pregnancy factor (EPF) has been repeatedly confirmed as indispensable to successful pregnancy. However, there is as yet little reported about how surgical abortion would affect the EPF activity, owing to the induced embryo loss. The aim of this study was to pursue this among a large number of patients available in the People's Republic of China.  METHOD: Sera from aborters were collected before surgical abortion and again on the 3rd, 5th and 7th day after treatment. EPF activity was detected by rosette inhibition assay.  RESULTS: Before surgical abortion, the mean level of EPF in pregnancy sera was about the same as that of the positive control. After surgical abortion, the EPF level declined rapidly for the first 3 days and then dropped gradually within the negative control range after 5–7 days.  DISCUSSION: Quantitative study of EPF activity along temporal dimensions (duration) due to surgical abortion further promotes the efficiency to take EPF activity and its rate of change as truly index for monitoring embryonic care and development of normal pregnancy.
The Immunosuppressive Cytokines Influence the Fetal Survival in Patients with Pregnancy‐Induced Hypertension
Authors: DEL GOBBO VERA , GIGANTI MARIA GABRIELLA , ZENOBI ROSSELLA , PREMROV MARIA GRAZIA , VILLANI VITTORIO
Published: December 2003

PROBLEM: The present study examines the hypothesis that the elevated levels of transforming growth factor (TGF)‐β1 and
interleukin (IL)‐10 would be protective for the fetus survival during
pregnancy‐induced hypertension (PIH). Moreover, we evaluate the IL‐12 and
IL‐15 serum concentrations and their relationships with PIH. 
METHOD OF STUDY: Serum samples were obtained before the onset of labor
from control and PIH groups. Cytokine concentrations were determined by
Enzyme‐Linked Immunoadsorbent Assay. 
RESULTS: Our data show that PIH women have significantly higher
TGF‐β1 and IL‐10 concentrations with respect to control
groups (P=0.0001). Similarly, macrophages from the PIH placentas
produce in vitro more elevated TGF‐β1 and IL‐10
levels compared to normal pregnant ones (P=0.02), also in the absence of LPS stimulation. IL‐12 and IL‐15 serum concentrations were not detectable in all pregnant groups.  CONCLUSION: We have found that PIH women have elevated concentrations of anti‐inflammatory/immunosuppressive cytokines, suggesting their important role in fetal allograft protection during the normal and pathological pregnancy.
Cervical Epidermal Growth Factor‐Receptor (EGF‐R) and Serum Insulin‐Like Growth Factor II (IGF‐II) Levels are Potential Markers for Cervical Cancer1
Authors: MATHUR SUBBI P. , MATHUR RAJESH S. , YOUNG ROGER C.
Published: December 2003

PROBLEM: To ascertain if cervical epithelial epidermal growth factor receptor (EGF‐R) and serum insulin‐like growth factor II (IGF‐II) levels are potential markers for cervical cancer.  METHOD OF STUDY: We tested cervical biopsies obtained from 18 controls, 3 women with cervical intraepithelial neoplasia (CIN) I, 17 women with CIN II and III, and 12 women with cervical cancer for EGF‐R using a quantitative immunofluorescent antibody assay. We measured serum IGF‐II levels using an enzyme‐linked immunosorbent assay in 20 controls, 26 CIN patients, 12 with cervical cancer before therapy, 5 with cervical cancer for <1 year, and 9 others ≥1 year after therapy.  RESULTS: The levels of cervical EGF‐R in women with CIN and cervical cancer were significantly higher (P<0.05 for CIN I;
P<0.001 for patients with CIN II and III or cervical cancer)
than in controls. Women with cervical cancer (P<0.001 vs.
controls) or advanced CIN (P=0.03) had elevated levels of serum
IGF‐II, while the women with CIN I had levels similar to controls. Women
with cervical cancer in the post‐therapy period had significantly lower
serum IGF‐II levels than the women with cervical cancer before therapy
(P<0.001).  CONCLUSION: Cervical epithelial EGF‐R and serum IGF‐II levels may be used for the diagnosis and prognosis of cervical cancer.
Stem Cell Factor (SCF) Concentrations in Peritoneal Fluid of Women with or without Endometriosis
Authors: OSUGA YUTAKA , KOGA KAORI , TSUTSUMI OSAMU , IGARASHI TOSHIO , OKAGAKI RYUGO , TAKAI YASUSHI , MATSUMI HIROTAKA , HIROI HISAHIKO , FUJIWARA TOSHIHIRO , MOMOEDA MIKIO , YANO TETSU , TAKETANI YUJI , TSUTSUMI OSAMU
Published: December 2003

PROBLEM: In the quest for possible involvement of stem cell factor (SCF), a cytokine known to have multiple effects, in the pathogenesis of endometriosis, we evaluated concentrations of SCF in peritoneal fluid (PF) of women with or without endometriosis.  METHOD OF STUDY: SCF concentrations in PF collected from women undergoing laparoscopy were measured, using a specific enzyme‐linked immunosorbent assay (ELISA). Reverse transcription‐polymerase chain reaction (RT‐PCR) analysis to detect gene expression of c‐kit, the receptor for SCF, was performed using the endometriotic tissue and the eutopic endometrium collected during the operation.  RESULTS: SCF concentrations in PF of women with endometriosis were significantly higher compared to women without endometriosis. Looking at SCF concentrations in PF of women with endometriosis stratified by disease stage, women with stage I and II exhibited relatively higher SCF levels in PF, whereas SCF levels in PF with stage III and IV were comparable with those without endometriosis. The expression of mRNA for c‐kit was detected in both the endometriotic tissue and the eutopic endometrium.  CONCLUSION: We demonstrated an elevation in SCF levels in PF associated with endometriosis and the presence of its receptor in endometriotic tissues. Given the known pleiotropic properties of SCF, the present results suggest that SCF might play a role in the pathogenesis of endometriosis.
Autoimmune Response in Women with Endometriosis
Authors: IBORRA A. , PALACIO J.R. , MARTÍNEZ P. , ULCOVA‐GALLOVA Z.
Published: December 2003

PROBLEM: The aim of this study was to investigate the humoral immune response to the female reproductive tissues associated with endometriosis (grades I–III) (n=52), compared with a group of healthy
fertile women (n=6).  METHOD OF STUDY: An ELISA with cultured endometrial cell lines in monolayer was used to determine the presence of anti‐endometrial antibodies (AEA). For anti‐zona pellucida antibodies (AZPA) assessment a conventional ELISA was employed. The presence of antibodies to human sperm (ASA) was performed by the tray agglutination test (TAT).  RESULTS: Endometriosis grade III was associated with AEA in serum in the 45.4% of patients. The presence of AEA in serum is correlated to endometriosis severity. The 8.7% of women with endometriosis showed ASA, and the 10.9% of them were positive for AZPA. Antibodies specific for endometrial cells do not show reaction to any gamete antigen (sperm or oocyte), suggesting that they are not cross reactive.  CONCLUSIONS: Severity of endometriosis is correlated with high titers of AEA.
Chaperonin 10 in the Rat Oocytes and Early Embryos: its Expression and Activity for Early Pregnancy Factor
Authors: KAWAMURA KAZUHIRO , FUKUDA JUN , KUMAGAI JIN , KUMAGAI AKIKO , TANAKA TOSHINOBU , ITOH HIDEAKI , ITO KAZUEI , KODAMA HIDEYA
Published: December 2003

PROBLEM: The aim of this study was to investigate the expression of chaperonin (cpn) 10 and cpn 60 mRNA in oocytes or embryos, and to further explore the possibility that early pregnancy factor (EPF) is identical with cpn 10.  METHOD OF STUDY: The expressions of cpn 10 and cpn 60 mRNA in oocytes and embryos at the different stages (1‐cell, 2‐cell, 8‐cell, and morula) were examined by polymerase chain reaction techniques. The EPF activity of native rat cpn 10 isolated from rat livers was evaluated by the rosette inhibition test.  RESULTS: Similar levels of mRNA of cpn 10 and cpn 60 were detected in oocytes and embryos at every stage. There were no detectable EPF activities in the native cpn 10. Immunoprecipitation using polyclonal antibodies against cpn 10 did not affect the activity of EPF in the pregnant rat serum.  CONCLUSION: Our results do not support the hypothesis that cpn 10 is identical with EPF.
Maternal Estrogen Receptor‐β Expression during Mouse Gestation
Authors: SPONG C.Y. , MCCUNE S.K. , STERNBERG E.M. , GUSTAFSSON J.A.
Published: December 2003

PROBLEM: Although estrogen receptor (ER)‐α has been well characterized, the recently identified novel ER‐β has not. In some tissues, there is overlap of the ERs, which allows for rescue in cases of deficiency; in other tissues, the ERs appear to have opposite effects. The objective of this study was to evaluate the expression of ER‐β during pregnancy.  METHOD OF STUDY: Pregnant mouse uteri (embryonic days 6–14, 16, 18) were studied. ER‐α and ER‐β oligonucleotide probes were end‐labeled and in situ hybridization histochemistry was performed.  RESULTS: ER‐β was strongly expressed in maternal ovaries; there was no other evidence of strong expression during gestation. ER‐α was expressed in the uterus throughout gestation, with decreasing intensity as gestation progressed, and in maternal ovarian tissue.  CONCLUSIONS: Differential expression of the two ERs was apparent during pregnancy, with ER‐α playing a dominant role. This may have implications for selective drug treatment targeting estrogen receptors.
Successful Uterine Steroid Therapy in a Case with a History of Ten Miscarriages
Authors: OGASAWARA MAYUMI , AOKI KOJI
Published: December 2003

A woman experienced ten unexplained first trimester miscarriages. Uterine steroid therapy was performed just before her 11th pregnancy. Prednisolone was inserted into the uteral cavity before ovulation and her 11th pregnancy resulted in success. Our experience in this case confirms the potential usefulness of this treatment for patients with unexplained recurrent miscarriages.
Controlled Ovarian Hyperstimulation – A State of Endothelial Activation1
Authors: ORVIETO RAOUL , SCHWARTZ ARIELA , BAR‐HAVA ITAI , ABIR RONIT , ASHKENAZI JACOB , LA‐MARCA ANTONIO , BEN‐RAFAEL ZION
Published: December 2003

PROBLEM: To aim of the study was to investigate whether controlled ovarian hyperstimulation (COH) causes endothelial activation and whether there is a correlation between endothelial activation and serum sex‐steroid levels.  METHOD OF STUDY: The study population consisted of 14 consecutive patients undergoing our routine IVF long gonadotropin‐releasing hormone‐analog protocol. Blood was drawn three times during the COH cycle: (1) day when adequate suppression was obtained (Day‐S); (2) on the day of or the day prior to hCG administration (Day‐hCG); and (3) on the day of ovum pick‐up (Day‐OPU). The levels of sex steroids and plasma soluble endothelial (E)‐selectin were compared among the time points. Soluble E‐selectin was measured with a commercial sandwich enzyme‐linked immunosorbent assay.  RESULTS: Soluble E‐selectin levels were significantly higher on Day‐OPU than Day‐S and Day‐hCG, whereas no difference was observed between Day‐hCG and Day‐S. No significant correlations were found between soluble E‐selectin level and patient age, number of gonadotropin ampoules used, number of oocytes retrieved, or serum estradiol, progesterone and human chorionic gonadotropin levels.  CONCLUSIONS: Human chorionic gonadotropin administration leads to endothelial activation regardless of the degree of ovarian response. Further studies are required to elucidate the relationship between COH and endothelial activation. These findings may lead to new strategies for predicting and preventing complications of COH, such as severe ovarian hyperstimulation syndrome.
Pregnancy Outcome and Ovarian Antibodies in Infertility Patients Undergoing Controlled Ovarian Hyperstimulation
Authors: LUBORSKY J. , PONG R.
Published: December 2003

PROBLEM: Several endocrine markers are well‐established, but not absolute, predictors of successful outcomes following controlled ovarian hyperstimulation. Another potential predictor for success may be a marker of ovarian autoimmunity. Ovarian antibodies (OVAB) are detected in women with unexplained infertility. We tested the hypothesis that women with OVAB have a poorer pregnancy outcome in in vitro fertilization (IVF). 
METHOD OF STUDY: Serum samples (n=47) were assessed by a previously described immunoassay for OVAB in a cross‐sectional, retrospective study design. 
RESULTS: Women who became pregnant had a lower frequency of OVAB than women who did not become pregnant (25.0% [4/16] vs. 58.1% [18/31], respectively; P=0.03). There was no significant difference in day 3 estradiol, amount of human menopausal gonadotropin given, peak estradiol, the number of follicles observed, or the number of eggs retrieved between women who achieved pregnancy and those who did not.  CONCLUSIONS: Together with other information such as reproductive hormone levels and measures of follicle growth, OVAB may contribute additional information for prediction of successful IVF outcomes.
Immunocontraception of White‐Tailed Deer with GnRH Vaccine
Authors: MILLER LOWELL A. , JOHNS BRAD E. , KILLIAN GARY J.
Published: December 2003

PROBLEM: Reduction of excess numbers of white‐tailed deer (Odocoileus virginianus) is an example of a potential use for immunocontraception as a means of wildlife population management.  METHOD OF STUDY: A 4 year multifaceted study was conducted to determine the long term effects of gonadotropin releasing hormone (GnRH) contraceptive vaccine on the fertility and behavior of female and male white‐tailed deer. Deer were monitored for breeding behavior, hormone levels, pregnancy, fawning and GnRH specific antibody levels.  RESULTS: Treatment lead to reduced fawning rates, altered estrus behavior, reduced concentrations of progesterone, contraception and failure to maintain pregnancy following conception. GnRH immunized does bred to untreated bucks had an 88% reduction in fawning caused by either immunocontraception or immunocontragestion. The vaccine effect is reversible, directly related to the antibody titer. Infertility lasted up to two years without boosting. GnRH immunized bucks demonstrated no interest in sexual activity when paired with control females. Depending on the immunization schedule, antlers either dropped early or remained in velvet.  CONCLUSIONS: The results of this study demonstrate that GnRH vaccine is effective in inducing a reversible infertility in white‐tailed deer, the infertility lasting up to two years without boosting.
Chemotactic Activity for Polymorphonuclear Leukocytes: Meconium Versus Meconium‐Stained Amniotic Fluid
Authors: YAMADA TETSUO , MATSUBARA SHIGEKI , MINAKAMI HISANORI , KOHMURA YASUHIRO , HIRATSUKA MITSUHIRO , SATO IKUO
Published: December 2003

PROBLEM: Our previous study indicated that meconium‐stained amniotic fluid (turbid AF) possessed a potent chemotactic activity for leukocytes, which may be dependent on interleukin‐8. It is not known, however, whether meconium itself possesses this chemotactic activity.  METHOD OF STUDY: Meconium samples were collected from mature neonates with and without turbid AF. A 5% meconium suspension in phosphate buffered saline was prepared and measured for its chemotactic activity for leukocytes using the blind well chamber technique. Concentrations of IL‐8, TNFα and IL‐1β were also measured with ELISA.  RESULTS: The number of leukocytes that migrated to the meconium suspension (35±27) was comparable with that of the clear AF (31±37), but was significantly lower than that of the turbid AF (184±62, P<0.0001). The meconium suspension contained much lower levels of IL‐8, TNFα and IL‐1β than the turbid AF. 
CONCLUSIONS: Meconium itself exhibits a lower chemotactic activity for polymorphonuclear leukocytes than turbid AF in vitro. The leukocyte chemotactic activity of turbid AF does not originate from meconium itself.
Chromosome Studies in Patients with Defective Reproductive Success
Authors: RADOJČIĆ BADOVINAC ANĐELKA , BURETIĆ‐TOMLJANOVIĆ ALENA , STARČEVIĆ NADA , KAPOVIĆ MILJENKO , VLASTELIĆ IVICA , RANDIĆ LJILJANA
Published: December 2003

PROBLEM: The objective of this study was to evaluate the contribution of chromosomal anomalies to decreased fertility in humans.  METHOD OF STUDY: In order to investigate the aetiology of infertility in our population and to assess the karyotype in a group of infertile couples and individuals with fertility problems, 782 persons (259 couples, 158 male and 106 female) with different clinical diagnoses of sterility and infertility were analysed cytogenetically.  RESULTS: The overall frequency of major chromosomal aberration was 13.1% (103/783), which suggests that fertility or sterility problems in this population are due to chromosomal aberrations. Couples experiencing repeated spontaneous abortions, having malformed children or having sterility problems had chromosomal abnormalities in 18.0% (47/259 couples) of the population studied, and constituted chromosomal disorders occured in couples seeking IVF and ICSI with prevalence of 22.2% (8/38 couples), especially minor mosaicism of sex chromosomes in the female partners. The prevalence of chromosome abnormalities in infertile men was 17.7% (28/158), and in subfertile females, it was 26.4% (28/106).  CONCLUSIONS: These results could indicate an increased tendency to miotic sex chromosome non‐disjuction in humans.
Anti‐Paternal Antibodies by Flow Cytometry in the Management of Alloimmunization on Recurrent Miscarriages
Authors: MATSUBAYASHI HIDEHIKO , IZUMI SHUN‐ICHIRO , SUGI TOSHITAKA , YOSHIKATA KIKUO , MAKINO TSUNEHISA , MARUYAMA TETSUO , OZAWA NOBUAKI , YOSHIMURA YASUNORI
Published: December 2003

PROBLEM: There is no reliable laboratory test available to diagnose immunologically mediated miscarriages. We investigated the clinical significance of maternal anti‐paternal leukocyte antibodies by flow cytometry after alloimmunization.  METHOD OF STUDY: The flow cytometry crossmatch (FCXM) was performed in 158 patients with a history of three or more unexplained first‐trimester miscarriages without live birth. After negative FCXM patients were immunized, subsequent pregnancy outcomes and FCXM results were followed.  RESULTS: Of 112 subsequent pregnancies, 83 of 100 (83.0%) FCXM‐positive patients after immunotherapy had successful pregnancy outcomes, whereas seven of 10 (70.0%) FCXM‐negative patients had miscarriages (P=0.0001). The percent live birth ratio was 2.77 (CI, 1.07–7.16; P=0.0001) for FCXM‐positive patients compared to FCXM‐negative patients. The calculated predictive value showed that 75.6% of FCXM‐negative patients would have subsequent miscarriages.  CONCLUSIONS: Positive FCXM is closely associated with successful pregnancy outcome following immunotherapy. We propose that FCXM might be included in the routine laboratory tests for the management of recurrent miscarriages.
Adverse Influence of Numbers of Previous Miscarriages on Results of Paternal Lymphocyte Immunization in Patients with Recurrent Spontaneous Abortions
Authors: KATANO KINUE , SUGIURA OGASAWARA MAYUMI , SUZUMORI KAORU , AOKI KOJI
Published: December 2003

PROBLEM: To determine whether an increase in the number of previous miscarriages in recurrent spontaneous abortion patients is a risk factor in subsequent pregnancies with paternal lymphocyte immunotherapy.  METHOD OF STUDY: Live birth rates with reference to previous abortion numbers in recurrent spontaneous abortion patients were statistically compared between paternal lymphocyte immunotherapy and control groups, the latter retrospectively researched using historical data before 1981 in our clinic.  RESULTS: The overall live birth rate was 73% (169/232) in the immunotherapy group, and 48% (47/97) in controls (P<0.05). According to previous abortion numbers, the rates were 77% (114/148) versus 55% (36/65) (P<0.05) for three previous abortions, 70% (40/57) versus 38% (8/21) (P<0.05) for four and 56% (15/27) versus 27% (3/11) (not significant) for five, in the study and control groups, respectively.  CONCLUSIONS: The results confirm the efficacy of paternal lymphocyte immunotherapy, but demonstrate that the success rate deteriorates with the number of previous miscarriages.
Ciprofloxacin Affects Pregnancy Loss in CBA/JxDBA/2J Mice Possibly via Elevation of Interleukin‐3 and Granulocyte Macrophage‐Colony Stimulating Factor Production
Authors: SAVION SHOSHANA , SHEPSHELOVICH JEANNE , TODER VLADIMIR , BLANK MIRI , SHOENFELD YEHUDA , FISHMAN PNINA
Published: December 2003

PROBLEM: The mechanisms mediating pregnancy loss are far from being understood, but it is believed that modulation of the maternal immune system, that is known to support pregnancy, might serve as a means for the treatment of habitual abortions. Thus, we examined the effect of the anti bacterial agent ciprofloxacin, which was shown to affect maternal immunoreactivity, on pregnancy loss in the resorption‐prone CBA/JxDBA/2J mouse combination as well as associated changes in Interleukin (IL)‐3 and Granulocyte macrophage‐colony stimulating factor (GM‐CSF) production.  METHOD OF STUDY: CBA/J females mated to DBA/2J males were treated with ciprofloxacin for 5 consecutive days. On day 15 of pregnancy, the number of resorbed embryos was recorded and IL‐3 mRNA expression as well as IL‐3 and GM‐CSF protein production by splenocytes were assayed by northern blotting and ELISA, respectively.  RESULTS: Ciprofloxacin treatment resulted in a significant reduction in the resorption rate as compared to the effect of the control antibiotic ceftazidime or PBS only, while not affecting the number of implantation sites/mouse. Also, splenocytes from ciprofloxacin‐treated CBA/J mice exhibited an increased level of IL‐3 mRNA transcripts as well as an elevation in IL‐3 and GM‐CSF protein production.  CONCLUSIONS: Our data demonstrate the ability of ciprofloxacin to reduce pregnancy loss in the CBA/JxDBA/2J mouse model, possibly via elevation of IL‐3 and GM‐CSF production.
Psychosocial Factors Regulating Natural‐Killer Cell Activity in Recurrent Spontaneous Abortions
Authors: HORI SHIRO , NAKANO YUMI , FURUKAWA TOSHIAKI A. , OGASAWARA MAYUMI , KATANO KINUE , AOKI KOJI , KITAMURA TOSHINORI
Published: December 2003

PROBLEM: The preconceptional natural‐killer cell (NK) activity predicts subsequent miscarriage among women with unexplained recurrent spontaneous abortion (RSA). Psycho‐neuro‐immuno‐endocrine network has recently been proposed as a mechanism for abortions. We therefore examined which psychosocial factors influenced the NK activity among women with RSA.  METHOD OF STUDY: We measured the preconceptional NK activity of 61 women with a history two consecutive unexplained first‐trimester miscarriages and no live births. We also administered semi‐structured interviews and a battery of self‐report questionnaires to assess their social support, personality, self‐esteem and psychiatric symptoms.  RESULTS: The preconceptional NK activity was negatively correlated with the women's neuroticism personality trait (r=−0.32, P=0.01) and current depressive symptoms (r=−0.26, P=0.05), and positively correlated with their self‐esteem (r=0.34, P=0.01).  CONCLUSIONS: In addition to several substances such as transforming‐growth‐factor beta and granulocyte‐macrophase colony‐stimulating factor, we found that low neuroticism, low depression scale score and high self‐esteem contributed to high NK activity among women with RSA.
Newly Exposed Immunochemically Cross‐Reactive Epitopes in Sperm‐Specific LDH After Glucosylation and Gossypol Interaction
Authors: GUPTA G.S. , SYAL NIDHI
Published: December 2003

PROBLEM: In a previous study, mouse lactate dehydrogenase‐C4 (LDH‐C4) after chemical modifications with gossypol (Gossy‐LDH‐C4) and glucosylation with lactose (Glu‐LDH‐C4) was found to induce high immunological infertility in allogenic mice. In the present study, the characterization of antibodies and cross reactivity of the antisera produced against Gossy‐ and Glu‐LDH‐C4 with purified somatic isozymes are being reported.  METHODS: Allo‐antisera generated in Balb/c mice (i.r. route) against one primary (50 μg) and two secondary doses (30×2 μg) in Al(OH)3 were tested for cross‐reactivity by ELISA and antibody avidity using Scatchard plot and Sip's plot.  RESULTS: Results suggested that IgG against native LDH‐C4 failed to recognize somatic isozyme, while antisera against chemically modified LDH‐C4 consistently reacted with purified LDH from kidney and placenta. Scatchard plots and antibody saturation curves of native and complexed LDH supported the presence of heterogenous antibodies with a mean association constant (Ka) of the order of 106−107 M−1, whereas diversity of heterogeneity, defined by diversity constant (a), ranged between 0.89 and 1.23. In general, anti‐Glu‐LDH‐C4 antiserum and native LDH‐C4. reacted with higher Ka (low affinity) with a diversity constant of 0.89 compared with interaction between native LDH‐C4 and it's antibodies.  CONCLUSIONS: It is concluded that LDH‐C4 is not an immunochemically sperm‐specific protein, in which crossreactive epitopes are hidden within its conformation. Due to the large intake of cotton seed (a source of gossypol) by cattle, its unrefined oil by humans in various parts of the world, and the prevelance of diabetic state all the world over, the present study warns of immunological consequences in situ following gossypol interaction and glucosylation of LDH and conformationally related proteins in circulation.
Female Sex Hormones Modulate the Function of LPS‐treated Macrophages
Authors: CHAO TZU‐CHIEH , CHAO HSIAO‐HSIANG , CHEN MIIN‐FU , GREAGER JOHN A. , WALTER ROBERT J.
Published: December 2003

PROBLEM: To study the effects of estradiol (E2) or progesterone on macrophage function in the presence of lipopolysaccharide (LPS).  METHOD OF STUDY: Male rat peritoneal macrophages were treated in vitro with 0.1 μg/mL of LPS and E2 or progesterone. 
RESULTS: At 10−2 ng/mL, E2 significantly (P<0.05; n=6) enhanced tumor necrosis factor (TNF) release by LPS‐treated macrophages. TNF release was significantly (P<0.05; n=6) inhibited by 102 ng/mL or 103 ng/mL of E2 and by progesterone at less than 10−3 ng/mL or greater than 10−1 ng/mL. E2 (10−4 and 10 ng/mL) and progesterone (10−6–10−4 ng/mL and 102 ng/mL) each significantly (P<0.05, n=8) enhanced H2O2 release by LPS‐treated macrophages. E2 (<10−2 and >10 ng/mL) and progesterone (10−7–104 ng/mL) each significantly inhibited (P<0.05; n=6) NO2− release by LPS‐treated macrophages.  CONCLUSIONS: Exposure to LPS tended to diminish the effects of E2 and to enhance the effects of progesterone on the parameters determined here. Such LPS‐associated alterations in the dose–response profile of macrophages to female sex hormones may contribute to gender‐related differences in the immune response under normal and pathological conditions.
Inhibition of the Antibody Responses to Rat Blood Transfusion Antigens in Mice by Boar Seminal Immunosuppressive Fraction
Authors: VESELSKÝ L. , JONÁKOVÁ V. , HOLÁN V. , DRAHORÁD J. , MAN ÁSKOVÁ P. , ŽELEZNÁ B. , DOSTÁL J. , VOBURKA Z.
Published: December 2003

PROBLEM: Immunosuppressive fraction of boar seminal vesicle fluid (ISF) was tested to muffle primary and secondary antibody responses to xenotranfusions. Contemporaneously, heparin non‐binding fraction of seminal plasma (H− fraction), presumed to be identical to ISF, was used to support the results.  METHOD: To study their similarity, ISF and H− fraction were analyzed by high‐performance liquid chromatography and the separated proteins by N‐terminal sequencing. In sera of mice treated with ISF or H− fraction, the productions of antibodies against rat erythrocytes and blood serum were evaluated by enzyme‐linked immunosorbent assay (ELISA). The productions of IgM, IgA, and IgG subclasses were followed by sandwich ELISA.  RESULTS: ISF and H− fraction were proved to be equal complexes of porcine seminal plasma (PSP) proteins PSP I and PSP II. Both inhibited antibody responses to rat erythrocytes and serum and the concentrations of IgM, IgG, IgG1 and IgG2 after the first transfusion with a long‐lasting effect. Both suppressed the secondary antibody production if applied before the second transfusion. IgA and IgG3 stayed uninfluenced. ISF and H− fraction had an equal immunosuppressive effect.  CONCLUSIONS: ISF was characterized biochemically, found to be identical to H− fraction, and determined to be powerful in overcoming unwanted exaggerated antibody responses to xenotransfusion.
Intravenous Immunoglobulin Therapy Influences T cell Adhesion to Extracellular Matrix in Women with a History of Recurrent Spontaneous Abortions
Authors: JERZAK MAL GORZATA , GÓRSKI ANDRZEJ , JERZAK MAL GORZATA , RECHBERGER TOMASZ , GÓRSKI ANDRZEJ
Published: December 2003

PROBLEM: To determine activated T cell adhesion level to extracellular matrix (ECM) in non‐pregnant women with a history of recurrent spontaneous abortions (RSA). In addition, to evaluate a small‐dose intravenous immunoglobulin (IVIG) therapy influences on T cell adhesion to ECM.  METHOD OF STUDY: Phytohemaglutinin (PHA) or phorbol myristate acetate (PMA) activated T cell adhesion to the following extracellular matrix proteins: collagen IV, fibronectin and elastin were studied in women with the history of RSA. In addition, IVIG immunotherapy influence on T cell adhesion was studied. Normal T cells adhesion values were established in non‐pregnant healthy women with the previous successful pregnancy outcome and in normal healthy pregnant women.  RESULTS: PHA activated T cell adhesion to collagen IV (P=0.04), fibronectin (P=0.0003) nad elastin (P=0.02) were significantly higher in women with the history of RSA when compared to non‐pregnant healthy women wit the previous successful pregancy outcome. IVIG immunotherapy normalized the T cell adhesion level and favored a successful pregnancy.  CONCLUSIONS: Increased T cell adhesion to collagen IV, fibronectin and elastin characterize women with the history of RSA. Decreased T cell adhesion to the main extracellular matrix components of human placenta may underlie possible effect of IVIG action.
Theoretical Basis for Herbal Medicines, Tokishakuyaku‐San and Sairei‐To, in the Treatment of Autoimmunity‐Related Recurrent Abortion by Correcting T Helper‐1/T Helper‐2 Balance
Authors: FUJII TOMOYUKI , KANAI TAKAO , KOZUMA SHIRO , HAMAI YOKO , HYODO HIRONOBU , YAMASHITA TAKAHIRO , MIKI AKINORI , UNNO NOBUYA , TAKETANI YUJI
Published: December 2003

PROBLEM AND METHOD OF STUDY: To get insight into the basis for the empirical usage of herbal medicines in the treatment of recurrent abortion, we examined whether Tokishakuyaku‐san (Toki) and Sairei‐to (Sai) modulate T helper‐1 (Th1) and T helper‐2 (Th2) cytokine release from peripheral blood mononuclear cells (PBMCs). The effects of these medicines were investigated as related to human leukocyte antigen (HLA)‐G, a non‐classical HLA class I antigen expressed on trophoblasts and a putative crucial player involved in fetomaternal immune interplay.  RESULTS: Toki and Sai increased the release of Th1 group cytokines, tumor necrosis factor (TNF)‐α and interferon (IFN)‐γ while preserving the inhibitory effect of HLA‐G on the release of these cytokines. As for Th2 group cytokine release, Toki was without effect in modulating interleukin (IL)‐4 release, regardless of the presence of HLA‐G, whereas Sai nullified the effect of the presence of HLA‐G to stimulate the release of IL‐4 without affecting its release in the absence of HLA‐G.  CONCLUSION: Toki and Sai may have therapeutic potential, particularly in autoimmunity‐related recurrent abortion where Th2 response is pathologically enhanced, but not in recurrent abortion involving alloimmune fetomaternal derangement, a condition of, rather, an enhanced Th1 response.
Enhanced Expression of Fas‐Associated Proteins in Decidual and Trophoblastic Tissues in Pregnancy‐Induced Hypertension
Authors: KOENIG JOYCE M. , CHEGINI NASSER
Published: December 2003

PROBLEM: To determine if feto‐placental tissues from gestations complicated by pregnancy‐induced hypertension (PIH) have altered expression of Fas‐associated proteins.  METHOD OF STUDY: The expression of several Fas‐related proteins was determined in fetal membranes, decidua, and placentas obtained from PIH‐affected (n=12, age range 32–36 weeks) and normal (n=6, age range 37–41 weeks) gestations. Paraffin‐embedded tissue sections were stained with specific monoclonal antibodies to Fas, Fas ligand (FasL), caspase‐3, and bax.  RESULTS: We observed greater expression of Fas and FasL in amnion and decidua from PIH‐affected gestations than in normal controls. Intense staining was observed only in the perivascular endothelium (caspase‐3) and in decidual cells (bax) from PIH gestations.  CONCLUSION: Differential expression of Fas‐related proteins in fetal membranes, decidua, and placentas from PIH‐affected gestations is consistent with increased apoptosis, and suggests activation of the Fas/FasL pathway in a tissue‐specific manner.
Identification of Epididymis‐Specific Antigens Using Neonatal Tolerization
Authors: KHOLE VRINDA , JOSHI SAURABH , SINGH SAPNA
Published: December 2003

PROBLEM: Conventional immunization using whole sperm containing multiple antigens as the immunogen followed by hybridoma technology usually gives antibodies to antigens invariably of testicular origin, probably because of the strong immunogenic nature of these antigens. Therefore, an alternate approach of neonatal tolerization or subtractive immunization has been utilized to raise antibodies specific to epididymis by suppressing immune response to testicular antigens.  METHOD OF STUDY: Neonatal mice were tolerized with testicular sperm proteins on days 0 and 5. These animals were then immunized with epididymal sperm proteins on day 21, followed by two boosters at biweekly intervals. Sera from these mice were used to localize epididymis‐specific antigens.  RESULTS: Sera from mice that were tolerized to testicular sperm proteins and later immunized with epididymal sperm proteins reacted only with epididymal proteins.  CONCLUSION: The results of this study demonstrate that neonatal tolerization with testicular sperm proteins, followed by immunization with epididymal sperm proteins, enhances the production of antibodies to proteins exclusively of epididymal origin.
Nitric Oxide (NO) and Interleukin‐1β (IL‐1β) in Follicular Fluid and Their Correlation With Fertilization and Embryo Cleavage
Authors: BARRIONUEVO MARCELO J. , SCHWANDT RENEE A. , RAO PAPINENI S. , GRAHAM LLOYD B. , MAISEL LAURA P. , YEKO TIMOTHY R.
Published: December 2003

PROBLEM: Using an IVF model, the goal of the study was to investigate the relationship between follicular fluid (ff) NO and IL‐1β levels, as well as their correlation with fertilization of mature oocytes and embryo cleavage rates.  METHOD OF STUDY: Follicular fluid was collected from 17 patients at the time of transvaginal oocyte retrieval following controlled ovarian stimulation. Oocytes harvested from these follicles were followed through fertilization and embryo cleavage. The NO metabolites nitrate/nitrite (NO3/NO2) were measured using the Griess reaction as an indirect assessment of NO activity. IL‐1β was measured using a high sensitivity ELISA system (Amersham, UK). The Student's t‐test was utilized for unpaired data with the means considered significantly different when P≤0.05. 
RESULTS: Follicular fluid NO3/NO2 levels were significantly lower in follicles containing mature oocytes that fertilized (n=30; 9.7±1.0 μM), versus those that did not fertilize (n=23; 15.4±2.4 μM; P<0.05). Follicles that contained oocytes that fertilized and went on to divide beyond the 6 cell stage had significantly lower ff levels of NO3/NO2 (n=18; 7.5±0.9 μM), as compared to ff that contained oocytes that did not fertilize or failed to develop beyond the 5 cell stage (n=35; 14.6±1.7 μM; P<0.01). No correlation was found between ff NO3/NO2 levels (n=28; 13.8±2.0 μM) and ff IL‐1β levels (n=28; 0.5±0.08 pg/mL). An analysis of ff IL‐1β levels in relation to fertilization and embryo cleavage rates revealed no correlation.  CONCLUSIONS: Lower ff NO3/NO2 levels at the time of oocyte retrieval are associated with adequate fertilization and embryo cleavage rates. In our IVF model, no correlation was found between ff IL‐1β levels and ff NO3/NO2, fertilization, or embryo cleavage rates.
Expression of Functional Chemokine Receptors of Human Placental Cells
Authors: ISHII MAKI , HAYAKAWA SATOSHI , ‐SUZUKI MIKI‐KARASAKI , CHISHIMA FUMIHISA , NAGAISHI MASAJI , SATOH KAZUO , HAYAKAWA SATOSHI , YOSHINO NAOTO , HONDA MITSUO , NISHINARITA SUSUMU
Published: December 2003

PROBLEM: Chemokine receptors of placental trophoblasts possibly act as co‐receptors or alternative receptors of maternal–fetal infection by HIV. To clarify their possible expression and the physiological roles of chemokines on human placentae, we studied chemokine/chemokine receptor expression and the effects of exogenous chemokines on choriocarcinoma cell lines.  MATERIALS AND METHODS: Placental samples were obtained from 13 placentae of various gestational ages. Villous tissue was mechanically dissected from samples. Trophoblasts were enriched by anti‐human chorionic gonadotropin (hCG)‐coated magnetic beads. Human choriocarcinoma cell lines (JAR, BeWo, JEG‐3) were maintained in RPMI 1640 media supplemented with 10% FCS. Expression of chemokine receptors was studied by RT‐PCR. The effects of MIP‐1α, RANTES, MCP‐1 on hCG production were estimated by EIA. Effects of chemokines on proliferation of choriocarcinoma cell lines were examined by MTT assay.  RESULTS: We observed mRNA expression of CCR‐1, 2, 3, 4, 5 and CXCR‐1, 2, 4 in 1st trimester placental villi, CCR‐1, 2, 4 and CXCR‐1, 2, 4 in 2nd trimester placental villi, CCR‐1, 2, 4 and CXCR‐4 in 3rd trimester placental villi. Using MACS enriched trophoblasts, we observed identical results. A choriocarcinoma cell line BeWo expressed CCR‐1, 3, 4 and CXCR‐1, 2, 4 while JEG‐3 and JAR expressed CCR‐1, 3, 4, 5 and CXCR‐1, 2, 4. Expression of the CCR‐5 and CXCR‐4 protein in choriocarcinoma cell lines and MACS‐enriched trophoblats were confirmed by flow cytometry. Chemokine MCP‐3, MIP‐1α, RANTES mRNA were expressed by the 1st, 2nd and 3rd trimester placental samples and the three choriocarcinoma cell lines examined. MCP‐1 was expressed by 1st and 2nd trimester placental villi. Administration of chemokines up‐regulated proliferation (10−1–10 ng/mL) and hCG production (10−1–10−2ng/mL) of the three choriocarcinoma cell lines examined.  CONCLUSIONS: Our results suggest possible roles of chemokines/chemokine receptors on placental physiology and their involvement in HIV transmission as alternative receptors.
The Production and Clinical Evaluation of Macrophage Colony‐Stimulating Factor and Macrophage Chemoattractant Protein‐1 in Human Follicular Fluids
Authors: KAWANO YASUSHI , KAWASAKI FUMIKO , NAKAMURA SATOMI , MATSUI NAOHIKO , NARAHARA HISASHI , MIYAKAWA ISAO
Published: December 2003

PROBLEM: In order to investigate the role of macrophage colony‐stimulating factor (M‐CSF) and macrophage chemoattractant protein‐1 (MCP‐1) in human ovulation, we measured the concentrations of M‐CSF and MCP‐1 in human follicular fluids (FFs) and correlated them with oocyte maturation.  METHOD OF STUDY: The oocytes were obtained from the FFs of 46 women undergoing in vitro fertilization and embryo transfer (IVF–ET). The concentrations of M‐CSF and MCP‐1 in the FFs were measured by enzyme‐linked immunosorbent assay. In addition, granulosa cells obtained from the FFs of IVF patients were cultured and treated with forskolin and 12‐O‐tetradecanoylphorbol 13‐acetate (TPA) for 24–48 hr. 
RESULTS: Concentrations of M‐CSF and MCP‐1 were significantly higher in the FFs than in the serum (P<0.01). M‐CSF concentrations tended to be higher, while MCP‐1 concentrations were significantly higher in the FFs containing mature oocytes than in FFs containing immature oocytes (P<0.05). The production of M‐CSF was markedly increased over the basal level after treatment with forskolin (10 μM) for 24 (P<0.02) and 48 hr (P<0.01); however, the production of MCP‐1 was unchanged.  CONCLUSIONS: Our data suggest that M‐CSF and MCP‐1 may play an important role in human preovulatory processes and that M‐CSF, in particular, may be regulated by cyclic adenosine monophosphate. M‐CSF and MCP‐1 may also be valuable biochemical markers in the evaluation of oocyte maturation.
TCRγδ+ T Lymphocytes in Unexplained Recurrent Spontaneous Abortions
Authors: PSARRA KATERINA , KAPSIMALI VIOLETTA , TARASSI KATERINA , ATHANASIADIS THEOFILOS , PAPASTERIADES CHRYSSA , DENDRINOS SPYROS , BOTSIS DEMETRIOS , KREATSAS GEORGIOS
Published: December 2003

PROBLEM: It is generally accepted that the immune system and cellular immunity in particular are involved in the mechanisms affecting the outcome of gestation. In order to evaluate a putative role of lymphocytes in the immunological mechanisms of unexplained recurrent spontaneous abortions (URSA), we studied peripheral blood lymphocyte subpopulations in 244 women with URSA and 44 controls.  METHOD OF STUDY: Direct immunofluorescence in whole blood with the appropriate combinations of monoclonal antibodies and flow cytometry was used.  RESULTS: The study showed: a) a statistically significant increase of the mean CD4/CD8 ratio (2.12±0.84 vs 1.85±0.63, P=0,039); b) a statistically significant decrease of the mean value of the percentage of CD5+CD19+ lymphocytes (0.4±0.6 vs 1.4±0.78, P<0.0001); and c) a statistically significant increase of the percentage of T lymphocytes expressing TCRγδ (4.68±3.19 vs 2.61±1.14, P<0.0001). It should be noted that a statistically significant high number of women with URSA (72/195, 36.9%) showed an increased percentage of TCRγδ T cells (≥5%, where 5 equals the mean value +2 standard deviations (SD) of the mean value of controls), whereas such a high percentage was not found in any control subject.  CONCLUSIONS: It seems that women who experienced URSA comprise a heterogeneous population, as far as immunological parameters are concerned. At least in a subgroup of them, TCRγδ+ T cells could be considered to play a role in the immune pathogenesis of fetal loss.
Involvement of μ‐Calpain in Human Sperm Capacitation for Fertilization
Authors: AOYAMA TOMOMI , OZAKI YASUHIKO , SUZUMORI KAORU , OZAKI YASUHIKO , KUNIMATSU MITOSHI , SASAKI MAKOTO , AOKI KOJI , TADA TOYOHIRO
Published: December 2003

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.
Integrin Pattern and Effect on Contraction in Cultured Testicular Peritubular Myoid Cells
Authors: MAGNANTI MASSIMO , GISMONDI ANGELA , GANDINI ORIETTA , MORRONE STEFANIA , ROSSI FABIO M. , SANTIEMMA VITTORIO , MICHETTI PAOLO M.
Published: December 2003

PROBLEM: Neither the integrin pattern nor the biological functions of integrins have been extensively documented in human cultured testicular peritubular myoid cells (TPMC). The integrin pattern and the presence of some proteins of the immunoglobulin superfamily on human TPMC as well as the role of integrins in TPMC contraction were examined.  METHOD OF STUDY: Integrin expression was evaluated by immunofluorescence and FACS analysis. To assess the role of integrin in TPMC contraction, human and rat cells were added to a collagen gel system and exposed to contractile stimuli.  RESULTS: The immunofluorescence and cytofluorimetric analysis showed that human cultured TPMC express α1, α2, α3, α5, α6, αv, β1, β3, and β4 integrin subunits, and significant amounts of intercellular adhesion molecule‐1 (ICAM‐1), whereas they do not present α4, β2, β7 subunits, nor intercellular adhesion molecule‐2 (ICAM‐2) and neural cell adhesion molecule (NCAM). The preincubation of human cells with an anti‐β1 mAb and of rat cells with a polyclonal anti‐β1 antibody inhibited TPMC contraction induced by different contractile stimuli.  CONCLUSION: Our investigation documented a broad integrin pattern on human cultured TPMC as well as a role for integrins in human and rat TPMC contraction.
Characterization of 80 kDa Human Sperm Antigen Responsible for Immunoinfertility
Authors: BANDIVDEKAR A.H. , VERNEKAR V.J. , MOODBIDRI S.B. , KOIDE S.S.
Published: December 2003

PROBLEM: An 80 kDa human sperm antigen (80 kDa HSA) has been identified by western blot technique using serum of an immunoinfertile woman as a probe. The 80 kDa HSA has been subsequently purified from sperm extract and investigated for antifertility effects.  METHOD OF STUDY: The purified 80 kDa HSA was used to immunize adult male and female rats. Rabbit anti 80 kDa HSA antibodies were used for immunofluorescent and immunohistochemical staining to demonstrate the presence of 80 kDa HSA on the sperm and to investigate its tissue distribution. The N‐terminal sequence of native 80 kDa HSA and the peptides obtained by its endoproteinase Lys‐C was determined.  RESULTS: Active immunization of male and female rats with 80 kDa HSA caused infertility in all the immunized animals. Immunofluorescent staining showed its localization on the head region of the human and rat spermatozoa. While immunohistochemical studies showed its localization in the testes and epididymis but not in other somatic tissues. Partial amino‐acid sequence analysis showed no sequence homology with any of the known protein in the database.  CONCLUSIONS: 80 kDa HSA is a promising candidate antigen for immunocontraception as it is characterized and found to cause infertility upon active immunization, specific to spermatozoa and is conserved.