bluuebunny/crossref_metadata_2025 huggingface dataset:
Vascular Cell Adhesion Molecule‐1 in Normal, Failed, and Ectopic Pregnancy
Authors: DANIEL YAIR , GAMZU RONI , LESSING JOSEPH B. , BAR‐AM AMIRAM , GEVA ELI , AMIT AMI , ESHED‐ENGLENDER TALMA
Published: December 2003

PROBLEM: Vascular cell adhesion molecule‐1 (VCAM‐1) is involved in early pregnancy establishment. This study sought to determine whether soluble VCAM‐1 (sVCAM‐1) serum levels differ among normal, failed, and ectopic pregnancy, its capacity to serve as a marker for pregnancy viability or ectopic pregnancy, and its correlation with serum progesterone and β human chorionic gonadotrophin (βHCG) levels. METHOD OF STUDY: Maternal serum samples were obtained from 20 women with ectopic, 10 with normal, and 10 with failed intra‐uterine pregnancy, all of comparable gestational age. Samples were assayed for sVCAM‐1, progesterone, and βHCG by specific assays. RESULTS: The median serum level of sVCAM‐1 was comparable between the three pregnancy types (normal: 578.3 ng/mL, range 434.4–699.5 ng/mL; failed: 567.8 ng/mL, range 401.9–669.5 ng/mL; and ectopic: 470.7ng/mL range, 328.2–1151.1 ng/mL). Serum levels sVCAM‐1 were not significantly correlated with βHCG or progesterone levels. CONCLUSION: sVCAM‐1 is not appropriate to serve as a marker for pregnancy viability or ectopic pregnancy.
Immunotherapy Prevents Recurrent Abortion Without Influencing Natural Killer Receptor Status
Authors: MIKI AKINORI , FUJII TOMOYUKI , HAMAI YOKO , YAMASHITA TAKAHIRO , KOZUMA SHIRO , TAKETANI YUJI , MIKI AKINORI , ISHIKAWA YOSHIHIDE , TADOKORO KENJI , JUJI TAKEO
Published: December 2003

PROBLEM: We assessed the expression of natural killer (NK) receptors in recurrent aborters before and after immunotherapy using their husbands’ peripheral blood mononuclear cells (PBMCs). METHOD OF STUDY: Using stored PBMCs from recurrent aborters before and after the immunotherapy, the expression of NK receptors, CD158a, CD158b, CD159 and CD94, were analyzed using monoclonal antibodies for respective receptors. The diversity of killer activatory receptors (KARs) and killer inhibitory receptors (KIRs) was also examined using reverse transcriptase‐polymerase chain reaction (RT‐PCR)‐single strand conformation polymorphism (SSCP) method. RESULTS: In recurrent aborters, no apparent changes in NK receptor expression and the balance between KARs and KIRs were found before and after the immunotherapy. CONCLUSION: The allo‐human leukocyte antigen (HLA)–stimulation caused by the immunotherapy for recurrent aborters did not affect the expression of NK receptors and the ratio of KARs to KIRs regardless of the outcome of subsequent pregnancies, suggesting that recurrent aborters may benefit from the immunotherapy through mechanisms unrelated to alteration in NK receptor status.
Effects of Paternal Lymphocyte Immunization on Peripheral Th1/Th2 Balance and TCR Vβ and VΓ Repertoire Usage of Patients with Recurrent Spontaneous Abortions
Authors: HAYAKAWA SATOSHI , KARASAKI‐SUZUKI MIKI , ISHII MAKI , KANAEDA TAKASHI , NAGAI NOBUHISA , TAKAHASHI‐YAMAMOTO NORIKO , TOCHIGI MEIJIN , CHISHIMA FUMIHISA , KIYOSHI FUJII TOM , SATOH KAZUO , ITOH TAKEYOSHI , OYAMA JUNKO , KITANAKA SUSUMU
Published: December 2003

PROBLEM: The mechanism of immunotherapy for patients with recurrent spontaneous abortions is not well understood. In order to investigate the suppressor mechanism of paternal lymphocyte immunization, we examined peripheral blood lymphocyte subpopulations and the repertoire of T‐cell receptor (TCR) gene segments. METHOD OF STUDY: Twelve patients with recurrent miscarriage were treated with immunization with paternal lymphocyte vaccinations three times during 12–14 weeks. Before and 2 weeks after the final inoculation, lymphocyte subsets and intra‐cellular interferon (IFN)‐Γ and/or interleukin (IL)‐4 production were examined by flow cytometry. TCR Vβ and VΓ repertoires were examined by semi‐quantitative reverse transcription‐polymerase chain reaction (RT‐PCR). RESULTS: We found no significant difference in CD4/CD8 ratios, prevalence of CD56+CD3+ or CD57+CD3+
cells (possible extrathymic T cells), ΓΔT cells, and
CD5+CD19+ (B‐1) cells. However, by in vitro
activation with phorbol 12‐mytistate 13‐acetate (PMA) and ionomycin,
peripheral CD4 cells demonstrated a significant decrease of
IFN‐Γ‐producing T helper 1 (Th1) cells and an increase of
IL‐4‐producing T helper 2 (Th2) cells after immunotherapy. Seven of nine
patients who exhibited remarkable decreases in Th1/Th2 ratios became
pregnant within 6 months after three courses of immunotherapy, and four
women delivered healthy babies, while none of the three patients who
exhibited an increased or unchanged Th1/Th2 ratio had full‐term
pregnancies (χ2<0.0001). Further, changes in usage of TCR Vβ and VΓ gene segments were observed after immunotherapy in six patients examined. CONCLUSION: Our findings suggest a shift of Th1‐dominant to Th2‐dominant status by vaccination might play important roles in maintaining successful pregnancies. Induction of some T cells that utilize different TCR repertoires possibly suppresses maternal rejection reactions.
Effect of Testicular Macrophage Conditioned Media from Rats with Autoimmune Orchitis on Leydig Cell Function
Authors: SUESCUN MARÍA O. , SUESCUN MARÍA O. , CALANDRA RICARDO S. , CALANDRA RICARDO S. , LUSTIG LIVIA
Published: December 2003

PROBLEM: The aim of this study was to investigate the influence of immune‐activated testicular macrophages obtained from rats with autoimmune orchitis (EAO) on Leydig cell steroidogenesis. METHOD OF STUDY: EAO was induced in rats by active immunization with testis homogenate and adjuvants. Testicular and peritoneal macrophages from rats with EAO were isolated and cultured for 24 hr. Testosterone (T) production by purified Leydig cells incubated in vitro with
macrophage‐conditioned media (CM) from rats with EAO or control rats was
measured.
RESULTS: An increase in T production by Leydig cells incubated with CM
from testicular, but not peritoneal, macrophages of rats with EAO was
observed. This increase was dose‐dependent up to a concentration of 30%
CM; proportions higher than 35% exhibited an inhibitory effect.
CONCLUSIONS: Immune‐activated testicular macrophages obtained from rats
with EAO induced both stimulatory and inhibiting steroidogenic effects on
Leydig cells in vitro and not the exclusively inhibitory action that has widely been attributed to activated macrophages. This dual effect probably depends on the ability of these cells to synthesize different molecules that may exert opposite effects.
Luteinizing Hormone Up‐Regulates the Expression of Interleukin‐1β mRNA in Human Granulosa‐Luteal Cells
Authors: CHEN HSIN‐FU , CHAO KUANG‐HAN , CHANG LI‐JUNG , HO HONG‐NERNG , YANG YU‐SHIH , SHEW JIN‐YUH
Published: December 2003

PROBLEM: Previously, we observed that follicular fluid obtained from patients with premature luteinization contained elevated interleukin‐1β (IL‐1β) levels. In this study, we aimed to examine the effects of luteinizing hormone (LH) on IL‐1β expression and IL‐1β on steroidogenesis in human granulosa‐luteal cells.  METHOD OF STUDY: Human granulosa‐luteal cells were obtained during oocyte retrieval. The cells were treated with either LH or IL‐1β and subsequently were examined for the level of IL‐1β transcript. The conditioned media were examined for IL‐1β protein and steroid hormone levels.  RESULTS: LH (250–500 mIU/mL) up‐regulated the expression of IL‐1β mRNA (up to a 4‐fold increase over control; P<0.05)
in the granulosa‐luteal cells. IL‐1β (5–50 ng/mL) increased
the basal, but not LH‐dependent, progesterone production from these cells
in a dose‐dependent manner after 96 and 144 hr of culture
(P<0.05). However, an inhibitory effect of IL‐1β on
LH‐dependent estradiol production was observed (up to 20% decrease,
P<0.05).  CONCLUSIONS: LH is capable of stimulating IL‐1β transcript expression in human granulosa‐luteal cells and may regulate ovarian steroidogenesis, at least partly through the activation of IL‐1β.
Anti‐Sperm Antibodies from Infertile Patients and their Cognate Sperm Antigens: A Review. Identity Between SAGA‐1, the H6‐3C4 Antigen, and CD52
Authors: DIEKMAN ALAN B. , NORTON ELIZABETH J. , WESTBROOK V. ANNE , KLOTZ KENNETH L. , NAABY‐HANSEN SØREN , HERR JOHN C.
Published: December 2003

PROBLEM: The correlation of anti‐sperm antibodies (ASA) with some instances of unexplained infertility implicates a role for these antibodies in blocking fertilization. Improved diagnosis and treatment of immunologic infertility, as well as a more complete understanding of the mechanism behind this phenomenon, are dependent on the identification and characterization of relevant sperm antigens.  METHOD OF STUDY: In this article, we review literature on methods employed to identify sperm antigens using anti‐sperm polyclonal and monoclonal antibodies from infertile patients and vasectomized men. Particular focus is given to approaches using human and mouse monoclonal antibodies to define the SAGA‐1 human sperm antigen.  RESULTS: ASA present in sera and genital tract secretions from infertile patients and vasectomized men have been employed in a variety of methods to identify sperm antigens. In an alternate approach, a monoclonal antibody (mAb), H6‐3C4, was immortalized from the lymphocytes of an infertile woman who exhibited sperm‐immobilizing titers. Subsequently, the sperm‐agglutinating, murine S19 mAb was shown to react with the H6‐3C4 cognate antigen. The H6‐3C4/S19 cognate antigen, designated Sperm Agglutination Antigen‐1 (SAGA‐1), was characterized as a polymorphic, highly acidic, GPI‐anchored glycoprotein on the surface of human spermatozoa. Purification with the S19 mAb followed by microsequencing demonstrated that the SAGA‐1 core peptide is identical to CD52, a glycoprotein on the surface of human lymphocytes. Immunoblot analysis demonstrated that these two glycoproteins differed in carbohydrate composition. Thus, sperm SAGA‐1 and lymphocyte CD52 represent glycoforms, glycoproteins with the same core peptide but with different carbohydrate structures.  CONCLUSIONS: Autoimmunity to the SAGA‐1 and/or CD52 glycoforms may lead to infertility. Structural and immunologic differences between these glycoproteins may be important factors in the etiology of immunologic infertility and other autoimmune disorders.
Polyherbal Formulations with Wide Spectrum Antimicrobial Activity Against Reproductive Tract Infections and Sexually Transmitted Pathogens
Authors: TALWAR G.P. , RAGHUVANSHI POONAM , MISHRA RACHNA , BANERJEE UMA , RATTAN ASHOK , WHALEY KEVIN J. , ACHILLES SHARON L. , ZEITLIN LARRY , BARRÉ‐SINOUSSI FRANÇOISE , DAVID ANNIE , DONCEL GUSTAVO F.
Published: December 2003

PROBLEM: Recent reports indicate high incidence of genital infections, most of which are sexually transmitted. Although specific drugs and antibiotics are available for some, a safe spermicidal formulation with wide spectrum antimicrobial action would be a desirable addition to the presently available spermicides.  METHODS: Formulations at different dilutions were tested in culture systems on standard strains and clinical isolates including some isolates resistant to drugs. The effect on (HSV)‐2 and Chlamydia trachomatis
was determined in vivo in progestin sensitized mice. The effect
on HIV‐1 was investigated in two standardized systems. 
RESULTS: Polyherbal cream inhibited the growth in culture of clinical
isolates of Candida albicans, Candida krusei and Candida
tropicalis. Both the polyherbal cream and the Praneem polyherbal
pessary inhibited urinary tract Escherichia coli (including
multidrug resistant strains), and Neisseria gonorrhoeae (including 2 strains resistant to penicillin). Both formulations manifested virucidal activity against HIV‐1 at>2 and 50% dilutions (in two different test systems) on contact for 1
–2 min. Intravaginal inoculation of the cream and the pessary
suspensions before inoculation of the pathogen prevented lesions and
vaginal transmission of HSV‐2 and C. trachomatis in progestin sensitized mice.  CONCLUSIONS: Polyherbal formulations have wide spectrum antibacterial, antifungal and antiviral effect against the tested sexually transmitted pathogens.
Interleukin‐1 Stimulates Human Uterine Prostaglandin Production Through Induction of Cyclooxygenase‐2 Expression
Authors: RAUK PHILLIP N. , CHIAO JYE‐PING
Published: December 2003

PROBLEM: Uterine infection occurs in as much as 20% of preterm labor and results in increased decidual cytokines. The objective of this study was to examine the effect of interleukin‐1 (IL‐1) and the cyclooxygenase‐2 (COX‐2) inhibitor, NS‐398, on myometrial prostaglandin (PG) production and COX‐2 expression.  METHOD OF STUDY: Human uterine myocytes were stimulated with IL‐1 (0–50 ng/mL) over 24 hr. PGE2, PGF2α,
and 6‐keto F1α were measured by enzyme‐linked immunosorbent assay. Both COX‐1 and COX‐2 proteins and mRNA were measured by western and northern blot, respectively.  RESULTS: IL‐1 increased PG production beginning at 6 hr. COX‐2 protein increased beginning at 4 hr and continued to increase at 24 hr. COX‐2 mRNA increased at 2 hr and peaked at 4 hr. NS‐398 blocked PG production but had no effect on COX‐2 protein or mRNA.  CONCLUSIONS: IL‐1 increases PG production by myometrium by increased COX‐2 expression. NS‐398 completely blocks IL‐1‐induced PG production. With intrauterine infection, IL‐1 may induce labor through the autocrine production of uterotonic PGs.
Do Soluble Cell Adhesion Molecules Play a Role in Endometriosis?
Authors: DANIEL YAIR , BARAM AMIRAM , FAIT GIDEON , LESSING JOSEPH B. , GEVA ELI , AMIT AMI , ESHED‐ENGLENDER TALMA
Published: December 2003

PROBLEM: Endometriosis is a chronic inflammatory disease associated with diverse immunologic disturbances. Cell adhesion molecules are essential for the development of immune and inflammatory reactions. This study was conducted to investigate whether or not serum and peritoneal levels of soluble cell adhesion molecules are altered in women with endometriosis.  METHOD OF STUDY: The study group comprised five women with moderate‐to‐severe endometriosis. Eight healthy women with a normal diagnostic laparoscopy served as controls. Serum and peritoneal fluid samples from both groups were analyzed for the soluble isoform of intercellular cell adhesion molecule‐1 (sICAM‐1), vascular cell adhesion molecule‐1 (sVCAM‐1), endothelial selectin (sES), and platelet selectin (sPS).  RESULTS: Serum levels of sICAM‐1 were significantly increased in women with endometriosis (median levels: 410.4 ng/mL; range: 233.9 ng/mL–598.4 ng/mL vs. 235.7 ng/mL; range: 187.4 ng/mL–323.7 ng/mL; P=0.02). Although the levels of sVCAM‐1, sES, and sPS in both samples were higher in the study group, the differences did not reach significance.  CONCLUSIONS: Our results suggest a role of ICAM‐1 in the pathophysiology of endometriosis. However, the role of other investigated cell adhesion molecules should be confirmed by further studies.
Establishment and Epitope Analysis of Allo‐Specific Cytotoxic T Lymphocytes at a Tumor Site Recognizing a Spouse's HLA‐A0206 Molecule
Authors: SEKI NAOKO , YAMADA AKIRA , SUEFUJI YUICHI , MINE TAKASHI , TANAKA SHOKO , GOMI SHIN‐YA , KAWAGOE NOBUTOSHI , KOUFUJI KIKUO , ITOH KYOGO
Published: December 2003

PROBLEM: The molecular basis of allo‐reactivity in reproductive immunity has not been fully clarified.  METHOD OF STUDY: Cytotoxic T lymphocytes (CTLs) were established from tumor‐infiltrating lymphocytes (TILs). The allo‐reactivity of the CTLs against various tumor cell lines or human leukocyte antigen (HLA)‐A allele‐transfected COS‐7 cells was measured by Cr‐release or interferon‐Γ production assay.  RESULTS AND CONCLUSIONS: We have established CTLs reacting to an HLA‐A0206 molecule that matched a spouse's HLA‐A allele from the TILs of a 68‐year‐old multiparous patient with gastric cancer. The amino acids at positions 66 and 88 in the α1 domain of HLA‐A0206, both of which were common in the other HLA‐A2 subtypes, were involved in the recognition by the CTLs. Endogenous peptides in the groove were not involved in the recognition. These results suggest the presence of long‐lasting memory CTLs raised by the reproduction process, and may facilitate a better understanding of the molecular basis of allo‐recognition during reproduction.
Study of Early Pregnancy Factor (EPF) in Equine (Equus caballus)
Authors: OHNUMA KATSUHIKO , YOKOO MASAKI , ITO KAZUEI , TAKAHASHI JUTARO , NAMBO YASUO , MIYAKE YOH‐ICHI , KOMATSU MAMORU
Published: December 2003

PROBLEM: Early pregnancy factor (EPF) is an immunosuppressive protein detected in the early pregnancy serum. We have already reported that we developed the rosette inhibition test for mare EPF and detected EPF in thoroughbreds. The aim of this study was to determine whether or not our method could be used clinically.  METHODS OF STUDY: The rosette inhibition test for equine EPF was carried out on serum from six nonpregnant and six pregnant Shetland ponies, a female and a male Chinese pony, and four nonpregnant and 13 pregnant thoroughbred mares. In the thoroughbreds, sera were collected during the pregnancy period. Furthermore, we measured progesterone and detected pregnant mare serum gonadotrophin (PMSG) in order to confirm pregnancy of the Chinese pony 3 and 6 months after mating.  RESULTS: In the nonpregnant Shetland ponies, the rosette inhibition titre (RIT) was 6.0±1.0 and EPF was negative. In contrast, in the pregnant ponies, the RIT was 9.2±0.4 and EPF was positive. Based on these results, we diagnosed pregnancy of the Chinese pony. The RIT of the female Chinese pony (3 months after mating) was above 10 and EPF was positive. Furthermore, we detected PMSG and progesterone in the serum of this pony. EPF appeared in the maternal blood circulation at 24–72 hr after mating, it was detected until the second trimester, and after that it disappeared from the maternal serum.  CONCLUSIONS: The pony's EPF was detected by using the same rosette inhibition test as in the thoroughbred and was present from 24 to 72 hr after mating until the second trimester. The results indicated that our method was useful for pregnancy diagnosis of Equine.
Preventive Effect of Monoclonal Antibodies to Intercellular Adhesion Molecule‐1 and Leukocyte Function‐Associate Antigen‐1 on Murine Spontaneous Fetal Resorption
Authors: TAKESHITA TOSHIYUKI , SATOMI MISAO , AKIRA SHIGEO , ARAKI TSUTOMU , NAKAGAWA YOKO , TAKAHASHI HIDEMI
Published: December 2003

PROBLEM: Are cell adhesion molecules involved in the murine model of immunologically‐mediated spontaneous abortion?  METHOD OF STUDY: Pregnant CBA/J female mice mated with DBA/2 male mice were injected with monoclonal antibodies (MAbs) to intercellular adhesion molecule‐1 (ICAM‐1) and leukocyte function‐associate antigen‐1 (LFA‐1). On day 13 of gestation, viable and resorbed embryos were counted. Natural killer (NK) cell activity in the spleen, mixed lymphocyte reactions (MLR), mixed lymphocyte‐placenta reactions (MLPR), and levels of interferon (IFN)‐Γ were assayed.  RESULTS: Significant suppression of fetal resorption was observed by the injection of MAb to ICAM‐1 and LFA‐1. NK cell activity and the MLR anti‐(CBA/J×DBA/2)F1 were reduced in the antibody‐treated CBA/J spleen. Moreover, the level of IFN‐Γ was significantly lower in the MLPR supernatants from the antibody‐treated group than those of the control group.  CONCLUSIONS: One mechanism in the murine model of spontaneous abortion may be through the interaction of cell adhesion molecules, which may modulate NK cell activities and cytokine production.
Evidence for Superantigen Involvement in Preeclampsia
Authors: LUPPI PATRIZIA , MATHIE THOMAS , FAAS SUSAN , RUDERT WILLIAM A. , TRUCCO MASSIMO , MCKNIGHT CRAIG , STEWART‐AKERS ANN M. , DELOIA JULIE A.
Published: December 2003

PROBLEM: Preeclampsia is the leading cause of maternal morbidity and premature fetal delivery in the United States, most likely involving the immune system in disease genesis. In this report, we tested the hypothesis that a superantigen phenomenon is an important factor in the pathogenesis of the disease.  METHOD OF STUDY: A semi‐quantitative polymerase chain reaction (PCR) was used to assess T‐cell receptor (TCR) β chain variable
(Vβ) regions as an indicator of T‐cell expansion in both
peripheral blood and basal plate of preeclamptic patients. All the
subjects were also molecularly typed to identify their HLA‐class II
alleles. 
RESULTS: In peripheral blood of the majority of the patients, there was a
high abundance of the Vβ4 gene family, which was not observed
in the control group. Polyclonality of this Vβ gene family
was confirmed by analysis of the Vα chain and the complementary determining region 3 (CDR3). The majority of patients carried the Human Leukocyte Antigens (HLA)‐DRB1*13 allele.  CONCLUSION: We present evidence for the existence of a superantigen‐like effect in at least a subset of patients with preeclampsia.
Fas Antigen Expression on the Decidual Lymphocytes of Pre‐Eclamptic Patients
Authors: DARMOCHWAL‐KOLARZ DOROTA , ROLINSKI JACEK , LESZCZYNSKA‐GORZELAK BOZENA , OLESZCZUK JAN
Published: December 2003

PROBLEM: Apoptosis has been proposed as a mechanism for maintaining the homeostasis in the immune system. Activated lymphocytes are removed by a programmed cell death process Fas/FasL‐mediated called activation induced cell death. The aim of the study was to investigate Fas antigen expression on decidual cells (T CD4+ lymphocytes, T
CD8+ lymphocytes and Natural Killer (NK) cells) of
pre‐eclamptic patients and healthy pregnant women. 
METHOD OF STUDY: 12 pre‐eclamptic patients and 10 healthy pregnant women
were studied. Lymphocytes were isolated from decidual tissues
mechanically, labeled by direct staining with monoclonal antibodies, and
analyzed using the flow cytometric method. 
RESULTS: We found Fas antigen expression on decidual NK cells and T
lymphocytes. CD 95 molecule expression and fluorescence intensity on NK
cells of pre‐eclamptic patients were lower when compared with controls
(P<0.05).  CONCLUSIONS: These findings suggest that decidual NK cells and T lymphocytes are able to undergo Fas/FasL‐mediated apoptosis. It seems that NK cells’ ability to undergo Fas/FasL‐mediated apoptosis in pre‐eclamptic patients can be altered because of lower CD95 molecule expression.
Mild Thyroid Abnormalities and Recurrent Spontaneous Abortion: Diagnostic and Therapeutical Approach1
Authors: VAQUERO ELENA , DE CAROLIS CATERINA , VALENSISE HERBERT , ROMANINI CARLO , LAZZARIN NATALIA , MORETTI COSTANZO
Published: December 2003

PROBLEM: The aim of this study is to evaluate the role of mild thyroid abnormalities in recurrent spontaneous abortion, and to assess the effects of two different therapeutical protocols.  METHOD: A prospective study in the population of recurrent aborters with mild thyroid abnormalities, evaluating the obstetric outcome in 42 patients. Sixteen thyroid autoantibodies positive patients were treated with thyroid replacement therapy, while 11 patients received intravenous immunoglobulins (IVIG). Fifteen patients, characterized by negative antithyroid antibodies, and having underlying thyroid pathology, were treated with thyroid replacement therapy.  RESULTS: Among patients with thyroid antibodies, 6 out of the 11 pregnancies (54.5%) treated with IVIG ended in live birth. In the thyroid supplementation group, 13 out of 16 pregnancies (81.2%) ended in live birth. Only one pregnancy loss occurred among patients with a mild underlying thyroid pathology treated with thyroid replacement therapy.  CONCLUSIONS: Mild thyroid abnormalities are associated with an increased rate of miscarriage. This poor obstetrical prognosis seems to be related to an impaired thyroid adaptation to pregnancy. Thyroid replacement therapy appears to be more effective than IVIG in preventing a new miscarriage.
Menstrual Cycle Dependent Fluctuations in NK and T‐Lymphocyte Subsets from Non‐Pregnant Human Endometrium
Authors: FLYNN L. , BYRNE B. , CARTON J. , O'FARRELLY C. , KELEHAN P. , O'HERLIHY C.
Published: December 2003

PROBLEM: To establish the relative proportions of endometrial lymphocyte subpopulations during the menstrual cycle.  METHOD OF STUDY: Lymphocytes were investigated by flow cytometry, during the early proliferative (EP), late proliferative (LP), early secretory (ES) and late secretory (LS) phases of the menstrual cycle.  RESULTS: Between the LP and LS phases, there was an increase in NK cells from 26.4% to 83.2% (P=0.0017) of the CD45+ population. T cells
decreased from 55.06% to 6.7% (P=0.0017). Within the T‐cell
population, CD8+ cells decreased significantly from 63.0% in the LP phase
to 54.2% in the LS phase (P=0.04). In contrast, potential
regulatory populations such as double negative (DN) T cells
CD3+CD4−CD8− and natrual T cells (NT) CD3+CD56+, increased
significantly in the LS phase (P=0.05; P=0.03). ΓΔ T cells, predominantly of the DN and NT phenotypes remained consistently low at all stages of the cycle (2.34%).  CONCLUSION: Endometrial lymphocyte fluctuations during the menstrual cycle may reflect hormonal regulation of maternal immunity, thereby promoting tolerance at the time of implantation.
Detection of Chlamydial Antigenic Material in Ovarian, Prostatic, Ectopic Pregnancy and Semen Samples of Culture‐Negative Subjects
Authors: TOTH M. , PATTON D.L. , CAMPBELL L.A. , CARRETTA E.I. , MOURADIAN J. , TOTH A. , SHEVCHUK M. , BAERGEN R. , LEDGER W.
Published: December 2003

PROBLEM: The pathogenesis of long‐term sequelae in Chlamydia
trachomatis infection is poorly understood. While serology indicates
previous chlamydial infection, culture studies are frequently negative.
We wanted to know whether in chronic cases the bacterium is absent or
persists in a dormant state where it evades detection. 
METHODS OF STUDY: Using immunoperoxidase (IP) staining and in situ
hybridization (ISH), we examined tissues of culture‐negative
subjects. 
Ovarian biopsy specimens from 19 culture‐negative women with pelvic
adhesions and/or tubal infertility were analyzed by both methods. Samples
of prostates from 10 culture‐negative men undergoing prostatectomy for
benign hypertrophy, two sets of semen samples from culture‐negative
sexual partners of 28 women with PID and/or bacterial vaginosis (BV), and
ten endometrium‐tube sample‐pairs from ectopic pregnancies (EPs) were
examined by IP only. 
RESULTS: Seven of the nineteen ovarian specimens tested positive for
Chlamydia antigen or deoxyribonucleic acid (DNA) (36%). Of the 10
hypertrophic prostates examined, 4 (40%) were positive. Of the 28 semen
samples examined, 10 (35%) tested positive. Tissue samples of 3 cases of
EP were positive by IP. 
CONCLUSIONS: 1. C. trachomatis antigen and nucleic acid can be frequently demonstrated in asymptomatic, culture‐negative men and women with chronic infection. 2. Chlamydia antigens may have an etiologic role in benign prostate hypertrophy and EP. 3. Antigenic material may be sexually transmissible. 4. IP and ISH identify temporarily inactive bacteria that may continue to act as immunostimulants and potentially reactivate as Chlamydia infection.
Early Pregnancy Factor is Required at Two Important Stages of Embryonic Development in the Mouse
Authors: ATHANASAS‐PLATSIS S. , CAVANAGH A.C. , MORTON H. , CORCORAN C.M. , KAYE P.L.
Published: December 2003

PROBLEM: The importance of early pregnancy factor (EPF) at the pre‐implantation stage of development (days 1–3 post‐coitum [p.c.]) has been previously established in this laboratory. However, the role of EPF at the implantation stage (days 4.5–5 p.c.) has not been determined. This present study therefore investigates the role of EPF at this important developmental stage, both in vivo and in
vitro. 
METHOD OF STUDY: Mated mice were passively immunized with anti‐EPF
antibodies at the peri‐implantation stage (days 3.5–4 p.c.) and
embryo implantation recorded. Parallel studies were conducted in
vitro, where the effect of anti‐EPF antibodies on trophoblast
outgrowth of blastocysts was determined. 
RESULTS: Administration of anti‐EPF antibodies in vivo at the peri‐implantation stage of development resulted in failure of embryos to implant. Similarly, trophoblastic outgrowth of blastocysts was adversely affected in the presence of anti‐EPF antibodies.  CONCLUSIONS: These results, together with previous findings that anti‐EPF antibodies retard embryonic development when administered at the early pre‐implantation stage, clearly demonstrate that EPF is required by the embryo at two important developmental stages – the one–two‐cell stage and the peri‐implantation stage.
Analysis of mRNA Levels for the MHC Class I‐Like Molecules CD1 and FcRn in Preimplantation Mouse Embryos
Authors: WARNER CAROL M. , PASCHETTO MIRIAM G.
Published: December 2003

PROBLEM: Major histocompatibility complex (MHC) antigens expressed on preimplantation embryos are important for the control of development, reproduction, and allo‐recognition of the embryo by the mother. Four types of MHC class I and MHC class I‐like antigens have recently been defined: class Ia, class Ib, class Ic, and class Id, based on their similar three‐dimensional protein structures. Class Ia and class Ib antigens are encoded in the MHC, whereas class Ic and class Id antigens are encoded by genes on other chromosomes. Both class Ia and class Ib MHC antigens are expressed on preimplantation mouse embryos. The function of the class Ia antigens on embryos is unknown, but the function of one class Ib antigen, Qa‐2, the product of the Ped gene, has been
found to control the rate of early cleavage division and subsequent
embryo survival. The expression of class Ic and class Id antigens on
preimplantation embryos has not yet been evaluated. In the present study,
we report the analysis of mRNA expression of two class Id genes,
CD1 and FcRn, in preimplantation mouse embryos. 
METHOD OF STUDY: A reverse transcription‐polymerase chain reaction
(RT‐PCR) assay was performed to analyze mRNA levels for CD1 and
FcRn in 1‐cell, 2‐cell, 8‐cell, and blastocyst stage embryos from
C57BL/6 mice. 
RESULTS: No expression of CD1 mRNA was found in any of the
preimplantation embryos tested. As a by‐product of this study, we found a
mistake in the published sequence of the mouse CD1 gene:
nucleotide 746 in the cDNA is a G not a C. This base change is in a site
recognized by the restriction enzyme PstI, thereby eliminating a
PstI cleavage site. Expression of mRNA for FcRn was found
in all preimplantation stages tested. Higher levels of mRNA for
FcRn were detectable in 2‐cell and 8‐cell embryos compared to
1‐cell and blastocyst stage embryos. 
CONCLUSION: This study shows that mRNA for FcRn but not for
CD1 is found in preimplantation mouse embryos.
Fas‐Mediated Apoptosis of Mouse Embryo Stem Cells: Its Role During Embryonic Development
Authors: ZOU GANG‐MING , REZNIKOFF‐ETIÉVANT MARIE‐FRANÇOISE , MILLIEZ JACQUES , LÉON ANNE , VERGÉ VÉRONIQUE , HIRSCH FRANÇOIS
Published: December 2003

PROBLEM: Fas antigen (APO‐1/CD95) can regulate the activity of various cells during adulthood. This study aimed at determining whether Fas may also be involved in the regulation of very early events such as the embryo preimplantation stage.  METHOD OF STUDY: We used mouse embryo stem (ES) cell line as a model for testing the effect of Fas crosslinking upon anti‐Fas monoclonal antibody (MoAb) treatment. In addition, this treatment was also applied to in‐vitro mouse‐embryo culture. 
RESULTS: Flow‐cytometry analysis of cultured ES cells demonstrated an
increase in Fas expression, unchanged in the presence of mouse
interleukin‐2, while greatly upregulated in the presence of
lipopolysaccharide (LPS). As determined by various means, ES cells may
undergo a Fas‐mediated apoptosis, slightly but significantly intensified
by the addition of LPS to cell cultures. We also report that anti‐Fas
MoAb directly inhibited two‐cell stage mouse‐embryo (preimplantation)
development in in‐vitro culture conditions.  CONCLUSION: These data suggest a novel mechanism controlling the regulation of physiological cell turnover as well as blastocyst implantation in early embryo development.
The Role of Amniotic Fluid Interleukin‐6, and Cell Adhesion Molecules, Intercellular Adhesion Molecule‐1 and Leukocyte Adhesion Molecule‐1, in Intra‐Amniotic Infection
Authors: HSU CHAUR‐DONG , AVERSA KRISTEN , MEADDOUGH ERIKA
Published: December 2003

PROBLEM: To determine amniotic fluid concentrations and correlations of interleukin‐6 (IL‐6), intercellular adhesion molecule‐1 (ICAM‐1), and leukocyte adhesion molecule‐1 (LAM‐1) in patients with and without intra‐amniotic infection.  METHOD OF STUDY: Fourteen specimens with intra‐amniotic infection and 45 without intra‐amniotic infection were studied. Intra‐amniotic infection was defined as the presence of a positive amniotic fluid culture. Amniotic fluid IL‐6, ICAM‐1, and LAM‐1 levels were determined by an enzyme‐linked immunoassay, and normalized by amniotic fluid creatinine levels.  RESULTS: Amniotic fluid concentrations of IL‐6 and LAM‐1 were significantly higher in patients with than without intra‐amniotic infection. However, amniotic fluid ICAM‐1 concentrations were not significantly different between two groups. Amniotic fluid IL‐6, LAM‐1, and ICAM‐1 were positively correlated.  CONCLUSIONS: Our data indicate that amniotic fluid IL‐6 is significantly associated with an increased adhesion molecule expression in intra‐amniotic infection. However, LAM‐1 plays a more important role than ICAM‐1 in intra‐amniotic infection.
Increased Apoptosis in Human Amnion is Associated with Labor at Term
Authors: HSU CHAUR‐DONG , MEADDOUGH ERIKA , BASHERRA HASAN , HARIRAH HASSAN , LU LI‐CHENG
Published: December 2003

PROBLEM: To characterize whether increased apoptosis in human amnion was associated with labor at term.  METHOD OF STUDY: Human amnion were obtained from term patients with vaginal delivery (n=5) or who underwent elective Cesarean section
(C/S) without labor (n=5). Apoptosis was performed by the TUNEL
(Terminal dUTP Nuclear End Labeling) assay. All nucleated cells stained
with propidium iodide in the amnion epithelial cells were identified in
red fluorescence. TUNEL positive apoptotic nuclei were identified in
green fluorescence. Five random fields of each specimen were blindly
counted by investigators. The percentage of apoptotic nuclei of total
nuclei (apoptotic index) was calculated and compared between the two
groups (25 microscopic fields for each group, respectively). 
RESULTS: Patients with term labor had a significantly higher mean
apoptotic index in amnion epithelial cells than that with elective C/S
without labor (27.3±4.1% versus 3.6±1.6%,
P<0.001).  CONCLUSIONS: Our data indicate that apoptosis in human amnion is significantly increased and associated with labor at term.
Fetal Endothelial Cells Express Vascular Cell Adhesion Molecule in the Setting of Chorioamnionitis
Authors: CRAVEN CATHERINE M. , WARD KEN , WARD KEN
Published: December 2003

PROBLEM: In intrauterine infection, inflammatory mediators may be released into the fetal circulation prior to fetal infection. We hypothesize that, in chorioamnionitis, inflammation alters fetal blood vessels. To test this, fetal endothelial cells were examined for vascular cell adhesion molecule (VCAM).  METHOD OF STUDY: Umbilical cords (n=9) from placentas with
chorioamnionitis were immunostained for VCAM. Controls from preterm
preeclamptic pregnancies (n=7) without histologic inflammation
were selected, and matched for gestational age and method of delivery.
VCAM sections were reviewed by a pathologist blinded to clinical
diagnoses. 
RESULTS: All endothelial cells from each of the nine cords from placentas
with chorioamnionitis had strong VCAM staining. Two of nine samples also
had acute cord vasculitis. No cord endothelial cells from preeclamptic
placentas demonstrated similar VCAM staining (p<0.01).  CONCLUSION: Histologic chorioamnionitis was associated with VCAM expression of the umbilical cord vessels. In chorioamnionitis, inflammatory mediators may have entered the fetal circulation to activate endothelial cells. Intrauterine inflammation was not restricted to the chorioamnion, but also involved the fetal circulation.
Intercellular Adhesion Molecule‐1 (ICAM‐1) in Cervicovaginal Fluid of Women Presenting with Preterm Labor: Predictive Value for Preterm Delivery
Authors: MARVIN KEITH W. , KEELAN JEFFREY A. , ZHOU REN LI , MITCHELL MURRAY D. , COLEMAN* MATTHEW A.G. , MCCOWAN LESLEY M.E. , COLEMAN MATTHEW A.G.
Published: December 2003

PROBLEM: Clinically useful tests for the prediction and diagnosis of preterm labor and delivery remain to be established. We have hypothesized that soluble intercellular adhesion molecule‐1 (sICAM‐1) in the cervicovaginal fluid of women with preterm labor may be a useful diagnostic tool.  METHOD OF STUDY: The cervicovaginal fluid of 103 women between 240 and 336 weeks gestation with preterm contractions and intact membranes was assayed for sICAM‐1.  RESULTS: Elevated sICAM‐1 concentrations predicted short intervals to delivery (area under receiver–operator characteristic (ROC) curves, 0.70–0.72 for delivery within 3, 7 and 10 days), with high specificity. Characteristics for delivery within 3 days at a 3 ng/mL threshold for a positive test were sensitivity 33.3%, specificity 98.9%, and positive and negative predictive values of 75.0% and 93.9%, respectively. Predictive ability was independent of and complementary to that of fetal fibronectin (fFN).  CONCLUSIONS: Measurement of sICAM‐1 in cervicovaginal fluid has potential as a predictor of preterm delivery in women with symptoms of preterm labor, particularly in conjunction with fFN testing.
Interleukin 8 Expression in Human Myometrium: Changes in Relation to Labor Onset and with Gestational Age
Authors: C.L. ELLIOTT , D.M. SLATER , W. DENNES , POSTON L. , P.R. BENNETT , L. POSTON
Published: December 2003

PROBLEM: Preterm labor remains the major cause of perinatal mortality and morbidity in normally formed babies. The mechanisms involved in the onset of preterm labor are poorly understood, mainly because the mechanisms initiating term labor remain ill‐defined.  METHOD OF STUDY: Human myometrial samples were collected at cesarean delivery from preterm (26–36 weeks gestation) and term (37–41 weeks) women. Women at term were either non‐laboring or laboring. The expressions of interleukin‐8 (IL‐8) mRNA and protein were measured by reverse transcriptase‐polymerase chain reaction and enzyme‐linked immunosorbent assay, respectively.  RESULTS: The expression of both IL‐8 mRNA and protein significantly increased in the term laboring group, compared with either the term non‐laboring or preterm groups. Levels of IL‐8 expression did not alter with advancing gestational age.  CONCLUSIONS: The increased expression of IL‐8 in laboring myometria at term supports the hypothesis that up‐regulation of IL‐8 has a role in the initiation of labor in association with an influx of neutrophils and the release of their collagenolytic enzymes into uterine tissues.
Production of Protein Hormones by Cultured Trophoblast Cells Isolated from Term and Early Placentae
Authors: MALEK ANTOINE , SAGER RUTH , WILLI AMARA , MÜLLER JÜRG , HÄNGGI WILLY , BERSINGER NIKLAUS , LEISER RUDOLF
Published: December 2003

PROBLEM: To compare the capacity of de novo hormone synthesis
by cultured trophoblast cells isolated from early and term placenta as
cytotrophoblast, and to determine the ability of these cells to
proliferate in culture. 
METHOD OF STUDY: Cytotrophoblast cells were isolated from term (TP,
38–42 weeks) and early placentae (EP, 8–13 weeks) by
enzymatic digestion and subsequent purification on a percoll gradient.
The net synthesis of the hormones human placental lactogen (hPL) and
human chorionic gonadotropin (hCG) was determined as the release during
culture+cell content after culture−cell content before culture.
Proliferation was determined using a dedicated colorimetric reagent
(CellTiter 96TM). 
RESULTS: Using a percoll gradient we were able to isolate three cell
bands with densities of 1.051, 1.058, and 1.063 g/mL, which were
predominantly cytotrophoblast cells as shown by immunocytochemical
analysis. The cytotrophoblast cells with the highest density (1.063 g/mL)
were used because they were found to release the highest amount of
hormones and have shown the lowest rate of cell death after 6 days in
culture. Both hCG and hPL showed different patterns of release during the
first 2–3 days of culture between TP and EP. While the release by
EP cytotrophoblast cells continued during 6 days of culture (n=4),
the concentrations for TP cytotrophoblast (n=4) reached a plateau
between 4 and 6 days. Net de novo synthesis calculated for
3×104 TP trophoblast cells cultured for 6 days
(mean±SD, n=4) was 8.65±9.05 mU for hCG and
0.95±0.45 ng for hPL. For EP, it was 395.5±265.5 mU for hCG
and 148.8±84.2 ng for hPL. Net synthesis of hCG was>10‐fold (TP) and>70‐fold (EP) higher than the initial cell content. While at term, hPL synthesis was only a fraction of the initial cell content, production by EP cytotrophoblast was 106 times the initial cell content. The extent of cell death after 6 days in culture was significantly (
P<0.02) higher for term (30–40%) than for early
trophoblast (10–20%). Using a proliferation detection agent during
the first 3 days of culture with first trimester cytotrophoblast cells,
we did not find any changes in the proliferative activity. 
CONCLUSIONS: There are differences in the functional activity between
trophoblast cells obtained from first and third trimester. The in
vitro findings are difficult to reconcile with the different patterns
of plasma concentrations of the two hormones observed in vivo during the course of pregnancy.
Transforming Growth Factor Beta Isoforms Production by Human Peritoneal Mesothelial Cells after Exposure to Hypoxia
Authors: SAED GHASSAN M. , ZHANG WENDY , DIAMOND MICHAEL P. , CHEGINI NASER , HOLMDAHL LENA
Published: December 2003

PROBLEM: Although human mesothelial cells (HMC) line nearly the entire abdominal cavity, little is known about their role in adhesion formation. This study determines the effect of hypoxia and transforming growth factor (TGF)‐β1 on the ability of HMC to produce TGF‐β1‐3, which have been implicated as mediators of the healing process.  METHOD OF STUDY: HMC were cultured under normal and hypoxic conditions, and treated with and without TGF‐β1 for 24 hr. RNA from each group was subjected to multiplex reverse transcription‐polymerase chain reaction to quantitate TGF‐β1‐3 mRNA levels.  RESULTS: Hypoxia resulted in 2‐ and 3.3‐fold increase, while TGF‐β1 treatment resulted in 1.4‐ and 1.2‐fold increase (normoxia) and 0‐ and 4.8‐fold increase (hypoxia) in TGF‐β1 and TGF‐β2 mRNA levels, respectively. There was no detectable TGF‐β3 mRNA in HMC before or after treatments.  CONCLUSION: TGF‐β1 treatment under hypoxia further extenuates endogenous TGF‐β2 but blocks TGF‐β1 production, thereby decreasing the TGF‐β1/TGF‐β2 ratio, which may result in the reduction of scarring and fibrosis.
Does Leptin Exhibit Cytokine‐Like Properties in Tissues of Pregnancy?
Authors: SOH E.B.E. , MITCHELL M.D. , KEELAN J.A.
Published: December 2003

PROBLEM: To determine whether leptin exhibits cytokine‐like properties in gestational tissues in light of its homologies with the class I family of cytokines.  METHOD OF STUDY: WISH and JEG3 cells, and amnion and choriodecidua explants, were treated inflammatory modulators (interleukin‐1β [IL‐1β], tumor necrosis factor‐α [TNF‐α] and bacterial lipopolysaccharide [LPS]) and leptin production was measured by immunoassay. Other agents known to regulate adipocyte leptin production were also tested for comparative purposes. In addition, WISH cells, JAR cells and placental explants were treated with leptin to assess its effects on production of IL‐8, IL‐6 and prostaglandin E2
(PGE2). 
RESULTS: Leptin production by all cells and tissues studied was
unaffected by treatment with IL‐1β (2.5 ng/mL), TNF‐α (25
ng/mL) and LPS (2.5 μg/mL). Dexamethasone stimulated leptin production
over two‐fold by WISH and JEG3 cells, whereas insulin also stimulated a
two‐fold increase in leptin production in JEG3 cells. IL‐6 production by
JAR cells and placental explants was stimulated (two‐ to three‐fold) by
leptin (300 ng/mL). PGE2 production was unaffected.  CONCLUSIONS: Leptin derived from gestational tissues is unlikely to play a role in inflammatory reactions within the placenta, but may regulate placental cytokine production. The physiological significance of amnion‐derived leptin remains to be established.
Altered Expressions of VEGF mRNA Splice Variants During Progression of Uterine‐Peritoneal Adhesions in the Rat
Authors: ROUT UJJWAL K. , OOMMEN KOSHY , DIAMOND MICHAEL P.
Published: December 2003

PROBLEM: Postoperative pelvic adhesions contribute to infertility, pelvic pain, bowel obstruction, and difficult reoperative procedures.  METHOD OF STUDY: In the present study, a rat uterine‐peritoneal adhesion model was developed to study the progression of adhesion formation during a course of 7 days following pelvic surgery. The distal 1 cm of each uterine horn and its adjacent peritoneum was abraded by six scratches with a scalpel blade, producing punctate bleeding. The scratched portion of uterine horn and the peritoneum was then held with Vicryl 3‐0 to promote adhesion. The uterine tissue and the portion of peritoneum, held with suture, were then excised from a group of four rats, each at 6, 12, 24, 48, 72 hr and 5 and 7 days following surgery. Total RNA was isolated from these tissues and the expression pattern of different splice variants of vascular endothelial growth factors (VEGF) was examined using relative abundance reverse transcriptase polymerase chain reaction (RA‐RT‐PCR) method.  RESULTS: Three known splice variants of VEGF mRNA (VEGF120,
VEGF164 and VEGF188), as well as an additional band
(∼510 bp), were amplified from these tissues. The relative abundance
of known VEGF isoforms demonstrated altered expression during adhesion
progression. When compared with noninjured uterine tissues,
VEGF120 and VEGF188 demonstrated up‐regulation
during early stages of adhesion formation, whereas VEGF164 rather demonstrated down‐regulation 24 and 48 hr following surgery.  CONCLUSIONS: The up‐regulation of VEGF isoforms during the progression of uterine‐peritoneal adhesion may be a compensatory mechanism regulating angiogenesis in order to provide nutrients and oxygen to the injured tissues.
Acute Intrauterine Infection Results in an Imbalance between Pro‐ and Anti‐Inflammatory Cytokines in the Pregnant Rabbit
Authors: K. LESLIE KIMBERLY , L. LEE SCOTT , M. WOODCOCK SUSAN , K. DAVIES JILL , L. ESKENS JOAN , S. GIBBS RONALD , HIRSCH EMMET , P. SHERMAN MICHAEL , S. MCDUFFIE JR ROBERT
Published: December 2003

PROBLEM: Intrauterine infection results in an increase in cytokines. This study compared the time courses for the pro‐ and anti‐inflammatory cytokine responses in 33 pregnant rabbits at 70% gestation. Pro‐inflammatory markers were activated nuclear factor‐kappa B (NF‐κB) in placenta and tumor necrosis factor‐alpha (TNF‐α) in amniotic fluid. These were compared to the anti‐inflammatory cytokine, interleukin‐1 receptor antagonist (IL‐1ra), in placenta and uterus.  METHOD OF STUDY: Does were endoscopically inoculated with Escherichia
coli through their cervices and sacrificed at six intervals between 0
and 30 hr post‐inoculation. 
RESULTS: Activated NF‐κB, determined by electromobility gel shift
assay, increased significantly 16 hr after bacterial inoculation (P≤0.05). This was directly mirrored by TNF‐α concentrations, determined by bioassay, in the amniotic fluid. However, IL‐1ra levels, determined by enzyme‐linked immunosorbent assay, did not increase in response to infection.  CONCLUSION: Intrauterine infection results in an imbalance between pro‐ and anti‐inflammatory cytokines that may potentiate infection‐induced preterm delivery.