bluuebunny/crossref_metadata_2025 huggingface dataset:
The Carriage of Pro‐Inflammatory Cytokine Gene Polymorphisms in Recurrent Pregnancy Loss
Authors: REID JANET G. , SIMPSON NIGEL A.B. , WALKER ROBERT G. , ECONOMIDOU OLGA , SHILLITO JANE , DUFFY SEAN R. , WALKER JAMES J. , GOOI H.‐C.
Published: December 2003

PROBLEM: Recurrent pregnancy loss (RPL) affects 2–4% of couples, and remains largely unexplained. Recent studies have examined the role of cytokines in the maintenance of normal pregnancy, which is linked with an increased expression of Th2 cytokines. Overexpression of Th1 cytokines is associated with RPL. Knowing that functional polymorphisms exist for certain cytokines, it has therefore been suggested that women with RPL may have a genetic predisposition to overexpress Th1 cytokines.  METHOD OF STUDY: The genes for interleukin‐1 beta (IL‐1β) and tumor necrosis factor alpha (TNF‐α) carry functional gene polymorphisms. In both cases these are biallelic polymorphisms that can be detected by polymerase chain reaction followed by restriction fragment length polymorphism. The aim of this pilot study was to assess whether carriage of the rarer alleles (TNF*2 and IL‐1B*2) could act as independent risk factors in recurrent miscarriage.  RESULTS: We found an increased incidence in the carriage of TNF*2, more pronounced in those women with two or more miscarriages. Carriage of the IL‐1B*2 either alone or in association with TNF*2 was not associated with recurrent miscarriage.  CONCLUSION: There may be a role for these cytokine gene polymorphisms in RPL.
TGFβ2 in Embryos with Inborn Anomalies: Effect of Maternal Immunopotentiation
Authors: IVNITSKY IRENA , TORCHINSKY ARKADY , SAVION SHOSHANA , SHEPSHELOVICH JEANNE , ORENSTEIN HASIDA , FEIN AMOS , TODER VLADIMIR
Published: December 2003

PROBLEM: TGFβs are among the main immunoregulatory molecules contributing to successful embryonic development. Besides, our and other studies revealed that maternal immunopotentiation has a potential to increase the resistance of the embryo to the teratogenic insult. This work was designed to evaluate: (1) whether the formation of teratogen‐induced anomalies is accompanied by an altered pattern of TGFβ2 expression in embryonic cells and (2) whether maternal immunopotentiation modifies the pattern of TGFβ2 expression in embryos responding to the teratogenic insult.  METHOD OF STUDY: Experiments were performed in embryos of ICR mice exposed to 15 and 40 mg/kg of a reference teratogen, cyclophosphamide (CP) on day 12 of gestation. A group of mice was immunopotentiated with xenogeneic rat splenocytes 21 hr before the beginning of mating. Embryos were examined for the occurrence of gross structural anomalies 24 and 72 hr after CP treatment. Then, immunohistohemistry and in situ hybrydization assays were used to evaluate the expression of TGFβ2 protein and mRNA in the brain, face, limbs and liver of these embryos.  RESULTS: No external anomalies were observed in embryos examined 24 hr after CP treatment. Embryos examined 72 hr after CP treatment at 40mg/kg exhibited agnathia, micrognathia, kinky tail, phocomelia, but no signs of dismorphogenesis were observed in the liver at the organ level. A significant increase in the expression of TGFβ2 mRNA was observed in cells, residing in the brain, face and limbs but not in the liver of CP‐exposed embryos tested 24 hr after CP injection in both doses. The level of TGFβ2 protein in these embryos did not differ from that of controls. In embryos tested 72 hr after CP injection in the high dose both TGFβ2 protein and mRNA expression were found to be elevated. Maternal immunopotentiation while enhancing the embryo's resistance to CP practically abolished an elevated expression of the TGFβ2 mRNA detected in tested organ structures of embryos of non‐immunopotentiated CP treated mice 24 hr after CP injection in both the low and the high doses. Also, a significant decrease in the level of TGFβ2 mRNA expression was observed in embryos of immunopotentiated mice examined 72 hr after CP treatment.  CONCLUSIONS: The results of this work show a possible involvement of TGFβ2 in the formation of teratogen‐induced structural anomalies and suggest that the stimulation of the maternal immune system may realize its protective effect by normalizing the level of TGFβ2 expression in teratogen‐targeted embryonic structures.
Inhibition of Murine Sperm‐Oolemma Binding by Antibodies to an Oocyte Membrane (OM) Antigen: Implication in Contraceptive Vaccine Development
Authors: NAZ RAJESH K. , LESLIE MONICA H. , ZHU XIAOLONG
Published: December 2003

PROBLEM: The present study was conducted to investigate the oocyte membrane protein(s) that is involved in sperm binding in the mouse, and whether or not it can be used for the development of a contraceptive vaccine.  METHOD OF STUDY: The zona‐free oocytes were treated with Triton X‐100 and the extract was analyzed for homogeneity in the sodium dodecylsulfate (SDS)‐polyacrylamide gel electrophoresis (PAGE) after staining with silver nitrate. The appropriate band of 50±4 kD, designated as OM antigen, was excised from the gel and used for the immunization. The female rabbits were immunized with the excised band per se and the female mice were immunized with the OM antigen after conjugation to keyhole limpet hemocyanin (KLH). The affinity‐purified antibodies were analyzed in the enzyme‐linked immunosorbent assay (ELISA), immunoprecipitation procedure, western blot procedure, indirect immunofluorescence technique (IFT), and sperm‐oolemma binding assay. Actively‐immunized mice were analyzed for in vivo fertility. 
RESULTS: The Triton X‐100 extract of zona‐free oocytes predominantly showed a single protein band of 50±4 kD in the SDS‐PAGE. Active immunization of female rabbits and of female mice with OM antigen raised high antibody titers (ELISA titer>1:4096) that specifically recognized the OM antigen in the immunoprecipitation and Western blot procedures, and reacted with the oocyte in the IFT. These antibodies demonstrated a significant (P<0.05) up to a complete block of sperm‐oolemma binding in the in vitro binding assay. Binding of both the acrosome‐intact and acrosome‐reacted sperm was inhibited. Mice actively immunized with OM antigen also showed a significant reduction in vivo fertility as seen by the 9‐day implants in uteri. Preliminary data indicate that the antibodies to OM antigen were tissue‐specific and did not react with the specific band in any tissue extract in the western blot procedure.  CONCLUSIONS: These results indicate that the OM antigen is involved in sperm‐oolemma binding and constitute an attractive molecule that needs further investigation for examining its utility in the contraceptive vaccine development.
Hypercoagulable Thrombophilic Defects and Hyperhomocysteinemia in Patients with Recurrent Pregnancy Loss
Authors: RAZIEL A. , FRIEDLER S. , SCHACHTER M. , RON‐EL R. , KORNBERG Y. , SELA B.A.
Published: December 2003

PROBLEM: Placental perfusion may be compromised by increased thrombosis that leads to pregnancy complications and recurrent pregnancy loss (RPL). Since heritable thrombophilic defects and hyperhomocysteinemia are associated with increased thrombosis, their prevalence was evaluated in RPL patients with special emphasis on combinations of the above pathologies.  METHODS OF STUDY: Evaluation of the prevalence of heritable thrombophilic defects (protein S, protein C, anti‐thrombin III deficiency, and the mutations for factor V Leiden, methylenetetrahydrofolate reductase [MTHFR], and prothrombin gene), hyperhomocysteinemia, and combinations of these pathologies in 36 non‐pregnant recurrent aborters compared with 40 parous women.  RESULTS: We found a relatively high prevalence of deficiencies of plasma coagulation proteins in RPL patients compared with the controls. A non‐significant different increase in factor V Leiden mutation was detected (6/36 [16%] compared with 2/40 [5%] in the control group, P=0.14]. Hyperhomocysteinemia was found in 31% of the RPL patients. MTHFR mutation homozygosity was found in 6/36 (16%) of the aborting patients. Combinations of hyperhomocysteinemia and MTHFR mutation were found in three patients, with folate deficiency in two patients, and with B12 deficiency in three.  CONCLUSIONS: Combinations of gene mutations, plasma protein deficiencies, and hyperhomocysteinemia, which are associated with an increased thrombotic risk, are more common in RPL patients compared with controls. Large‐scale prevalence studies are needed in order to draw conclusions as to the causative relation of such a condition and RPL.
The Association Between Anti‐Thyroid Antibodies and Pregnancy Loss
Authors: MATALON S. T. , BLANK M. , ORNOY A. , SHOENFELD Y.
Published: December 2003

The incidence of thyroid autoantibodies in women with recurrent fetal loss, infertility or women who miscarried appears to be increased compared with controls of reproductive age without previous abortions. There are few working hypotheses concerning the asociation between anti‐thyroid antibodies and the increased risk for pregnancy loss. The first hypothesis suggests that women with high titers of anti thyroid antibodies have underlying very mild thyroid “under function”. Another theory views the anti‐thyroid antibodies as simply secondary markers of a predisposition of autoimmune disease rather than the actual cause of pregnancy loss. An evolutionary explanation suggests that reproductive problems in women with high titers of autoantibodies exist in order to prevent the transmission of autoimmune genes to the next generation. The reason for the association between pregnancy loss and thyroid immunity is still not clear. The working hypotheses above supply multi factorial explanations which could act together, may even be in synergy, as the propulsion for the pregnancy loss. Until today the mechanism by which anti Tg are responsible for pregnancy loss is not clear. Induced animal models with high titier of anti Tg could provide direct evidence for the pathogenic role of anti‐thyroid antibodies and the mechanism that is responsible for the pregnancy loss in autoimmune thyroiditis. In the current presentation we describe data concerning the association between anti thyroid antibodies and pregnancy loss, and hypothesis which explain this association.
Differential Expression and Regulation of Inducible Nitric Oxide Synthase (iNOS) mRNA in Human Trophoblasts in vitro1
Authors: HAMBARTSOUMIAN E. , SRIVASTAVA R.K. , SEIBEL M.M. , HAMBARTSOUMIAN E. , SEIBEL M.M.
Published: December 2003

OBJECTIVES: To determine the expression of inducible nitric oxide synthase (iNOS) in human trophoblast and to examine the possible regulation of iNOS gene by cytokines.  MATERIALS AND METHODS: Total RNA was isolated from: 1) homogenized placental tissue; from 2) isolated and purified cytotrophoblast cells; and 3) cytotrophoblast and syncytiotrophoblast cells treated with cytokines in vitro. RNA was reverse transcribed and amplified by polymerase chain reaction, using specific primers for iNOS. Trophoblast cells were treated in vitro by interferon‐γ (IFN‐γ) in a dose of 10 ng/mL, Interleukin 1β (IL‐1β) (4 ng/mL) and leukemia inhibitory factor (LIF) (1 ng/mL). Trophoblasts were also subjected to immunocytochemistry using iNOS‐specific antibody to detect iNOS protein expression in these cells. 
RESULTS: The expression of iNOS mRNA was found both in placental tissue and isolated cytotrophoblast cells. In culture, the highly differentiated syncytiotrophoblast expressed more mRNA than cytotrophoblast cells. IFN‐γ and LIF, but not IL‐1β, induced iNOS mRNA expression in trophoblast cells in vitro. The effects of these cytokines on iNOS mRNA were only observed in syncytiotrophoblast cells, but not in cytotrophoblast cells. Immunocytochemical staining confirmed the trophoblast cells as a major source of the iNOS synthase production. 
CONCLUSIONS: 1) Human trophoblast cells are able to express the iNOS mRNA, hence suggesting a role for NO in placental growth and function. 2) LIF and IFN‐γ, but not IL‐1β, induce the iNOS mRNA expression in syncytiotrophoblast cells in vitro, suggesting possible similar regulatory mechanisms in vivo. 3) This study, for the first time, demonstrates the stimulating effect of LIF on iNOS gene expression in human tissue.
Comparative Effect of Danazol and a GnRH Agonist on Monocyte Chemotactic Protein‐1 Expression by Endometriotic Cells
Authors: JOLICOEUR CHRISTINE , LEMAY ANDRÉ , AKOUM ALI
Published: December 2003

PROBLEM: Endometriosis is associated with a chronic inflammatory process, and the increased number of activated peritoneal macrophages is one of the major hallmarks of this process. The medical treatment of the disease, which is based on the creation of an hypoestrogenic milieu unfavorable to the growth of endometriotic lesions, is often associated with a reduced peritoneal inflammation. The aim of this study was to investigate the ability of current therapeutic agents to modulate, through a direct mechanism, the expression by endometriotic cells of monocyte chemotactic protein‐1 (MCP‐1), a chemokine endowed with the potent faculty of recruiting and activating macrophages.  METHOD OF STUDY: Cells were stimulated with interleukin‐1 beta (IL‐1β) to induce MCP‐1 expression. MCP‐1 protein secretion and mRNA steady‐state levels were evaluated by ELISA and northern blot, respectively.  RESULTS: Our results show that danazol concentrations (10−7–10−5 M), taking into account the therapeutic levels found in the plasma of treated patients, inhibited MCP‐1 protein and mRNA steady‐state levels in endometriotic cells, whereas buserelin acetate (0.1–10 ng/mL), a GnRH agonist, had no significant effect. Dexamethasone, an anti‐inflammatory glucocorticoid, used at concentrations varying between 10−12 and 10−6 M, also displayed a dose‐dependent inhibitory action.  CONCLUSIONS: These results put into prominence the capability of danazol to directly inhibit the expression of a potent monocyte chemotactic and activating factor by ectopic endometrial cells shedding more light on the mechanisms underlying the clinical effects of hormonal therapeutic agents used in the treatment of endometriosis.
Human Leukocyte Antigen‐G‐Expressing Cells Differently Modulate the Release of Cytokines from Mononuclear Cells Present in the Decidua Versus Peripheral Blood
Authors: KANAI TAKAO , FUJII TOMOYUKI , UNNO NOBUYA , YAMASHITA TAKAHIRO , HYODO HIRONOBU , MIKI AKINORI , HAMAI YOKO , KOZUMA SHIRO , TAKETANI YUJI
Published: December 2003

PROBLEM: To better understand the role of human leukocyte antigen (HLA)‐G in regulating the T helper (Th)1/Th2 cytokine balance, one of key conditions in determining the fate of pregnancy, we asked whether the presence of HLA‐G protein altered the release of cytokines from both decidual mononuclear cells and peripheral blood mononuclear cells (PBMCs).  METHOD OF STUDY: The amounts of cytokines released from decidual mononuclear cells and PBMCs were compared in the presence or absence of HLA‐G‐expressing cells.  RESULTS: When cocultured with HLA‐G‐expressing cells, the amounts of tumor necrosis factor‐α and interferon‐γ released from decidual mononuclear cells and PBMCs were decreased, while the amounts of interleukin (IL)‐4 from PBMCs was increased, with IL‐4 release from decidual mononuclear cells being unchanged.  CONCLUSIONS: Upon contact with HLA‐G, decidual mononuclear cells, and PBMCs as well, modulate their ability to release cytokines in a way that may shift the Th1/Th2 balance towards relative Th2 dominance, suggesting a role for HLA‐G in maintaining pregnancy.
Influence of Breast‐feeding on the Production of Cytokines
Authors: SHIMAOKA YUKI , HIDAKA YOH , TADA HISATO , TAKEOKA KEIKO , AMINO NOBUYUKI , MORIMOTO YASUHIKO
Published: December 2003

PROBLEM: Recently, we reported increases in the production of interferon‐γ (IFN‐γ), interleukin‐2 (IL‐2), and IL‐4 during the postpartum period. The present study was undertaken to investigate whether these increases might be explained by increased prolactin while breast‐feeding.  METHOD: Whole blood from 41 women who were breast‐feeding, 13 women not breast‐feeding, and 31 healthy non‐pregnant women was stimulated with phorbol 12‐myristate 13‐acetate and ionomycin, and the levels of cytokines in the supernatant were measured by enzyme‐linked immunosorbent assay. Their serum levels of prolactin were measured by enzyme immunoassay.  RESULTS: Increases in IFN‐γ, IL‐2, IL‐4, and IL‐10 production were observed in women who were breast‐feeding but not in women who were not breast‐feeding. Serum levels of prolactin correlated with the levels of IFN‐γ in culture supernatant.  CONCLUSIONS: These results suggest that breast‐feeding induces production of cytokines and that IFN‐γ production is enhanced by physiological concentrations of prolactin.
HLA‐G Expression by Tumors
Authors: DAVIES B. , HIBY S. , GARDNER L. , LOKE Y.W. , KING A.
Published: December 2003

PROBLEM: It has been proposed that the expression of the non‐classical MHC class I antigen, HLA‐G, by trophoblast is one mechanism by which the placenta evades attack by maternal uterine NK cells. A similar mechanism is thought to be operative in the escape from immunosurveillance by tumor cells. However, data on the expression of HLA‐G by tumor cells are highly conflicting.  METHOD OF STUDY: In the present study, we have examined tissue sections from a wide variety of tumors by immunohistology and also several cell lines by flow cytometry and RT‐PCR. Whilst very faint bands were detected in three cell lines (hepG2, Mead, CaSki) by RT‐PCR, no tumors or cell lines were observed to express HLA‐G protein. Furthermore, we found that tumor deposits are not usually infiltrated by NK cells.  CONCLUSION: Our observations, therefore, do not support the proposal that tumor cells express HLA‐G in order to evade host NK cell cytolysis.
Immunologic Detection of Placental Lactogen During Pregnancy in a Mouse Model: A Preliminary Report
Authors: PATTEN KATHRYN F. , MATHUR SUBBI PILLAI , ROUDEBUSH WILLIAM E.
Published: December 2003

PROBLEM: To establish a rapid test differentiating ectopic from failed intrauterine gestation, using placental lactogen (PL) as a marker for placental cells.  METHOD OF STUDY: Sixteen Swiss Webster mice had synchronized ovulation and were mated. Eight mice were unmated controls. Study and control mice were sacrificed at 5, 7, 9, and 11 days gestation. Uterine sections were tested for PL by immunofluorescent antibody assay, enzyme‐linked immunosorbent assay, and dot blot analysis. Human endometrial samples from a missed abortion and a nonpregnant woman were also tested.  RESULTS: Placental lactogen was detectable only in pregnant uterine samples (placental cells only) by all assays and was absent in the endometial glandular cells of nonpregnant uterine samples.  CONCLUSION: Three methods detected placental lactogen in the pregnant mouse and human tissue. This is the first step towards developing a reliable clinical test for human endometrial samples from early pregnancy to differentiate early abortion from ectopic pregnancy.
Localization and Distribution of Platelet Activating Factor Receptors in the Mouse Ovary and Oviduct During the Estrous Cycle and Early Pregnancy
Authors: LASH GENDIE E. , LEGGE MICHAEL
Published: December 2003

PROBLEM: This study determined platelet activating factor (PAF) binding to ovaries and oviduct tissues to identify the location of PAF–tissue interaction.  METHOD: Mouse ovaries and oviducts taken during the estrous cycle and days 1, 4 and 7 post‐conception (pc) were analyzed by [
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H]PAF binding using frozen sections and autoradiography. 
RESULTS: For the outer epithelium of estrous and metestrous ovaries there was no significant difference in PAF binding (P>0.05); however, both stages were significantly different from proestrous (P<0.001). Ovarian stroma PAF binding was significantly higher (P<0.001) at estrous than metestrous, with no PAF binding at proestrous. Binding of PAF to estrous ovarian follicles was significantly greater (P<0.001) than at proestrous and metestrous. Estrous oviduct binding of PAF was significantly increased for stroma, inner epithelium and lumen (P<0.001 in all cases). At days 1 and 7 pc, all ovarian tissues had the greatest PAF binding with day 4 pc failing to bind PAF except for a significant decrease in corpora luteal binding (P<0.001). Oviduct binding of PAF was greatest at day 1 pc.  CONCLUSION: Ovarian and oviduct PAF receptor expression corresponds to peak embryo PAF synthesis and establishes the basis for PAF–PAF receptor mediated early pregnancy signaling system.
Cytokine Production by Natural Killer Lymphocytes in Follicular and Luteal Phase of the Ovarian Cycle in Humans
Authors: BOUMAN A. , MOES H. , HEINEMAN M.J. , DE LEIJ L.F.M.H. , FAAS M.M.
Published: December 2003

PROBLEM: The aim of this study was to test the hypothesis that, during luteal phase of the ovarian cycle, as compared with follicular phase, the cytokine productive capacity of peripheral natural killer (NK)‐lymphocytes in humans is shifted towards a “Th2‐type”‐like response.  METHOD OF STUDY: Intracellular Th1 and Th2 cytokine production by in vitro activated peripheral NK‐lymphocytes in a whole blood preparation of the follicular and the luteal phase of the ovarian cycle was measured by flow cytometry.  RESULTS: There was no difference in interferon (IFN)(‐γ, interleukin (IL)‐2, IL‐4, and IL‐10 cytokine production in activated NK‐lymphocytes when comparing luteal phase with follicular phase of the ovarian cycle. However, there was a significant increase in peripheral NK‐lymphocyte number in luteal phase compared with follicular phase.  CONCLUSION: The cytokine productive capacity of peripheral NK‐lymphocytes is not shifted towards a “Th2‐type”‐like response in the luteal phase as compared with the follicular phase of the ovarian cycle in humans.
Immunohistochemical Staining of IL‐1 Alpha and IL‐1 Receptor Antagonist but not IL‐1 Beta in Cultures of Sertoli Cells
Authors: HULEIHEL M. , ZEYSE D. , BECK M. , HULEIHEL M. , ZEYSE D. , LUNENFELD E. , POTASHNIK G. , MAZOR M. , PRINSLOO I. , HULEIHEL M. , ZEYSE D. , LUNENFELD E. , BECK M. , POTASHNIK G. , MAZOR M. , ZEYSE D. , BECK M.
Published: December 2003

The interleukin‐1 (IL‐1) system has been suggested to be involved in the cell–cell cross talk within the testis. To identify a testicular cell source of IL‐1 alpha, IL‐1 beta and IL‐1 receptor antagonist (IL‐1ra), immature mouse Sertoli cells were isolated, purified, cultured and examined for the cellular compartment localization of these cytokines by immunohistochemical staining. Our results show that both Germ cells and Sertoli cells in unpurified Sertoli cell cultures (before hypotonic shock) and purified culture of Sertoli cells (after hypotonic shock) were stained for IL‐1 alpha. The levels of this cytokine were increased in Sertoli cells when the purified cultures were stimulated with lipopolysaccharide (LPS) (5 μg/mL). However, we could not identify a positive staining for IL‐1 beta when Sertoli cell cultures were stained for this cytokine, even after stimulation with various concentrations of LPS (0.1–10 μg/mL). On the other hand, immunohistochemical staining of isolated Sertoli cells without treatment with hypotonic shock (cultures containing Sertoli cells and Germ cells) for IL‐1ra showed constitutive positive staining of both cell types (Sertoli cells and Germ cells). Our results, using immunohistochemical staining, may indicate the different expression of IL‐1 alpha, IL‐1 beta and IL‐1ra in Sertoli cells. These results may suggest the involvement of IL‐1 system in the autocrine and paracrine regulation of testicular cell functions.
Interleukin‐1β‐induced Prostaglandin E2 Production in Human Myometrial Cells: Role of a Pertussis Toxin‐sensitive Component
Authors: HERTELENDY FRANK , RASTOGI PRERNA , MOLNÁR MIKLÓS , ROMERO ROBERTO
Published: December 2003

PROBLEM: The objective of this study was to evaluate the possible role of pertussis toxin (PTX)‐sensitive G‐protein(s) in interleukin‐1β (IL‐1) signaling in human myometrial cells (HMC).  METHOD: Primary cultures of HMC were stimulated with human recombinant IL‐1 alone or in combination with PTX. Prostaglandin (PG) E2 in the medium was measured by radioimmunoassay, cyclooxygenase type 2 (Cox‐2) and IκB by western analysis, and the activities of two members of the mitogen‐activated protein kinase (MAPK) family of enzymes, ERK‐2 and JNK, by the phosphorylation of appropriate substrates.  RESULTS: IL‐1 increased PGE2 output during an 18‐hr long incubation by 21.7‐fold (n=5 experiments). This increase was inhibited by 57% after pretreatment overnight with PTX. IL‐1‐induced expression of Cox‐2 protein was also suppressed to a similar degree in PTX‐treated HMC cultures. Degradation of the nuclear factor kappa B (NF‐κB)‐inhibiting protein (IκB), a critical step in IL‐1 signaling to the nucleus, was significantly inhibited by PTX, as was IL‐1‐induced activation of ERK‐2 and JNK.  CONCLUSIONS: It is suggested that the occupied IL‐1 receptor‐generated signal in HMC is transmitted by multiple pathways. One is coupled to a PTX‐sensitive G‐protein upstream from the MAPK phosphorylation cascade. This, in turn, may interact with another signaling pathway, the activation of NF‐κB, via the phosphorylation of the IκB kinase complex.
Interleukin‐6 Up‐Regulates the Oxytocin Receptor in Cultured Uterine Smooth Muscle Cells
Authors: RAUK PHILLIP N. , FRIEBE‐HOFFMANN ULRIKE , CHIAO JYE‐PING , RAUK PHILLIP N. , WINEBRENNER LIZA D.
Published: December 2003

PROBLEM: Intra‐uterine infection results in the production of inflammatory cytokines, including interleukin‐6 (IL‐6). Increased oxytocin‐receptor (OTR) concentrations are associated with the onset of preterm labor. We hypothesize that infection up‐regulates OTR expression through IL‐6‐induced transcription factors.  METHOD OF STUDY: Primary cultures of human myometrium were treated for various time periods or with different concentrations of IL‐6 and OTR mRNA as well as OTR binding were measured by means of reverse transcription polymerase chain reaction and
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I‐ornithine‐vasotocin‐binding assay. To study underlying mechanisms of OTR changes with IL‐6 treated, cells were also incubated with genistein or H7 (tyrosine and serine phosphorylation inhibitors), respectively.  RESULTS: OTR mRNA increased 2.5‐fold after 4 hr of IL‐6 treatment and OTR binding 1.4‐fold after 8 hr of cytokine stimulation. The IL‐6‐induced increase in binding was blocked by genistein and H7.  CONCLUSIONS: IL‐6 up‐regulates uterine OTR mRNA expression and binding capacity in cultured human myocytes most likely through tyrosine and serine phosphorylation pathways involving the nuclear factor STAT‐3.
Soluble Endothelial and Platelet Selectins in Serum and Ascitic Fluid of Women with Ovarian Hyperstimulation Syndrome
Authors: DANIEL YAIR , GEVA ELI , BAR‐AM AMIRAM , LESSING JOSEPH B. , AMIT AMI , GEVA ELI , ESHED‐ENGLENDER TALMA
Published: December 2003

PROBLEM: The selectins, a group of cell adhesion molecules, are major mediators of inflammatory, immunologic, and angiogenic reactions. Their possible involvement and levels of the soluble isoform in serum and ascitic fluid of women with ovarian hyperstimulation syndrome (OHSS) were not previously evaluated.  METHOD OF STUDY: This prospective, case‐control study involved 16 women with OHSS. Ten matched women treated by controlled ovarian stimulation and eight healthy women with normal diagnostic laparoscopy served as controls. Serum and peritoneal fluid samples obtained from all subjects were assayed for soluble endothelial selectin (sES) and soluble platelet selectin (sPS) by specific enzyme‐linked immunosorbent assay.  RESULTS: Significantly higher levels of sES (median, 17.1 [9–41] vs 9.3 [2.4–21.3] ng/mL [P=0.03]) and sPS (median, 23 [13–277] vs 6.5 [1.6–28.7] ng/mL [P=0.001]) were observed in the peritoneal fluid of women with OHSS than the basal levels of healthy, non‐stimulated women. Women with OHSS had significantly lower sES serum levels than those treated by controlled ovarian hyperstimulation without OHSS, while their sPS serum levels were comparable.  CONCLUSIONS: Asitic fluid of women with OHSS contains appreciable amounts of soluble selectins, suggesting their ovarian origin and possible involvement in the syndrome.
Association between Interleukin Concentration in Follicular Fluid and Intracytoplasmic Sperm Injection (ICSI) Outcome
Authors: HAMMADEH M.E. , ERTAN A.K. , BALTES S. , BRAEMERT B. , ROSENBAUM P. , SCHMIDT W. , GEORG T.
Published: December 2003

PROBLEM: The aim of this study was to determine the presence and concentration of interleukin IL‐6, IL‐8, and granulocyte‐macrophage‐colony‐stimulating factor (GM‐CSF) in pre‐ovulatory ovarian follicular fluid (FF) of patients undergoing controlled ovarian hyperstimulation for intracytoplasmic sperm injection (ICSI) therapy on one hand, and to find out whether these cytotokine concentrations could be used as a predictive parameter for ICSI outcome.  DESIGN: The levels of IL‐6, IL‐8, and GM‐CSF were measured from women that underwent ICSI therapy and the results were compared between the patients who became pregnant after ICSI and those who did not.  MATERIAL AND METHODS: FF was obtained from 84 women participating in our ICSI program. The patients underwent ovarian hyperstimulation with human menopausal gonadotropin after they had been treated with gonadotropin‐releasing hormone agonist (GnRH‐a) for pituitary down‐regulation. The concentration of IL‐6, IL‐8, and GM‐CSF was measured by the highly sensitive enzyme‐linked immunosorbent assay method.  RESULTS: Twenty‐eight women became pregnant (group I) and the other 56 (group II) did not. The main concentrations of IL‐6, IL‐8, and GM‐CSF in FF of patients who became pregnant were 5.4±2.9, 207.9±118.3, and 1.01±0.8 pg/mL, respectively, and the corresponding values in patients who did not become pregnant were 7.0±5.1, 230.8±183, and 1.34±0.09 pg/mL, respectively. No statistical difference was found between the two groups with regard to the age of patients, IL concentration in FF, mean number of retrieved oocytes, mean number of mature oocytes (metaphase II), and the fertilization rate.  CONCLUSION: IL‐6, IL‐8, and GM‐CSF were found in most FF investigated either from patients who became pregnant after ICSI treatment or from those who did not. Furthermore, no significant differences were found between the IL‐6, IL‐8, and GM‐CSF concentrations in both groups. The mean oocyte retrieval, fertilization, and cleavage rates were not significant between the patients who became pregnant and those who did not. Therefore, the IL‐6, IL‐8, and GM‐CSF concentrations in pre‐ovulatory FF cannot be used as a reliable prognostic index for fertilization ability, developing potential, and implantation probability.
Polymorphism of Human Leukocyte Antigen‐E Gene in the Japanese Population with or without Recurrent Abortion
Authors: KANAI TAKAO , FUJII TOMOYUKI , YAMASHITA TAKAHIRO , HYODO HIRONOBU , MIKI AKINORI , UNNO NOBUYA , KOZUMA SHIRO , TAKETANI YUJI , KEICHO NAOTO , TOKUNAGA KATSUSHI
Published: December 2003

PROBLEM: The aim of this study was to investigate the gene frequencies and shared alleles of human leukocyte antigen (HLA)‐E gene in Japanese couples with or without recurrent abortion.  METHOD OF STUDY: Polymerase chain reaction (PCR)‐single strand conformation polymorphism (SSCP) analysis was carried out to detect polymorphism in exon 3 of the HLA‐E gene in 30 Japanese couples with recurrent abortion and 38 normal Japanese couples with proven fertility.  RESULTS: No point mutation was detected in exon 3 of HLA‐E in both recurrent aborters and normal controls. HLA‐EG and HLA‐ER alleles were detected with frequencies of 66.7% and 33.3% in couples with recurrent abortion and 69.2% and 30.8% in normal couples, respectively. The gene frequency of HLA‐EG was higher than that of HLA‐ER, which is contrary to that found in Caucasian, African–American and Hispanic people but similar to Chinese people. The frequency of each allele was not significantly different between recurrent aborters and normal controls. The number of shared alleles between each couple with recurrent abortion is not significantly different from that with normal controls.  CONCLUSION: Allele frequencies of HLA‐E were suggested to be different in Asian people from those in other ethnic people. In light of no specific distribution pattern in recurrent aborters, HLA‐E polymorphism does not seem to play a role in the pathogenesis of recurrent abortion.
Pregnancy Outcome in Recurrent Spontaneous Abortion Associated with Antiphospholipid Antibodies: A Comparative Study of Intravenous Immunoglobulin Versus Prednisone Plus Low‐Dose Aspirin
Authors: VAQUERO ELENA , VALENSISE HERBERT , MENGHINI SIMONA , DI PIERRO GIUSEPPE , ROMANINI CARLO , LAZZARIN NATALIA , CESA FABRIZIO
Published: December 2003

PROBLEM: To compare the use of intravenous immunoglobulins (IVIG) with prednisone plus low‐dose aspirin (LDA) in treating pregnant women with a history of recurrent fetal loss having the antiphospholipid antibody (aPL), in terms of live‐birth rate and maternal and perinatal morbidity.  METHOD: A prospective, two‐centers trial study included 82 recurrent aborters with aPL syndrome. Twenty‐nine were treated with prednisone and LDA in one center, 53 received IVIG in the other center. Maternal and fetal outcomes and pregnancy complications were compared between groups.  RESULTS: Live‐birth rates were equivalent between groups (78 vs 76%). Mean birth weight was higher in the IVIG group than in the prednisone plus LDA group. In the prednisone‐ plus LDA‐treated patients, gestational hypertension and gestational diabetes were found significantly more often than in the IVIG‐treated group (14 vs 5% and 14 vs 5%, respectively).  CONCLUSION: In patients with aPL syndrome, IVIG treatment improved pregnancy outcome, with significantly lower pregnancy complication rates, when compared with prednisone plus LDA therapy.
Serum Levels of Inhibin B, Unlike Inhibin A and Activin A, Are Not Altered in Women with Preeclampsia
Authors: YAIR DANIEL , ESHED‐ENGLENDER TALMA , KUPFERMINC MICHAEL J. , GEVA ELI , FRENKEL JENNY , SHERMAN DAN
Published: December 2003

PROBLEM: To investigate whether inhibin A, inhibin B, and activin A serum levels are altered in women with preeclampsia.  METHOD OF STUDY: Serum samples of 20 women with preeclampsia (study group) and 20 normotensive pregnant women, matched for maternal and gestational age and parity, were assayed for inhibin A, inhibin B and activin A by specific enzyme‐linked immunosorbent assay.  RESULTS: Median serum concentrations of inhibin A and activin A were significantly higher among women with preeclampsia than in women with normotensive pregnancies, while inhibin B levels were comparable in both groups. Activin A levels were positively correlated with those of inhibins A and B, and inhibin A levels were positively correlated with diastolic blood pressure and inhibin B concentration in the study group.  CONCLUSIONS: Inhibin A and activin A, but not inhibin B, serum levels are markedly increased in women with preeclampsia. These hormones might serve as an endocrine marker for preeclampsia.
Correlation Between Decreased Type‐II Interleukin‐1 Receptor and Increased Monocyte Chemotactic Protein‐1 Expression in the Endometrium of Women with Endometriosis
Authors: KHARFI ABDELAZIZ , AKOUM ALI
Published: December 2003

PROBLEM: Monocyte chemotactic protein‐1 (MCP‐1), a potent inducer of macrophage recruitment and activation, is overexpressed in the eutopic endometrium of women with endometriosis. Eutopic endometrial cells of women with endometriosis secrete higher levels of MCP‐1 than those of normal women, following stimulation with interleukin‐1 (IL‐1). The aim of this study was to examine whether there is any correlation between the expression of IL‐1 receptor type II (IL‐1RII), a specific downregulator of IL‐1 activity, and that of MCP‐1 in the eutopic endometrium of women with endometriosis.  METHOD OF STUDY: Endometrial biopsies of 46 women with endometriosis and 22 healthy women were evaluated simultaneously for IL‐1RII and MCP‐1 expression by immunohistochemistry.  RESULTS: Our study revealed a highly significant correlation between the decreased expression of IL‐1RII and the increased expression of MCP‐1 in the endometrial tissue of women with endometriosis (Spearman correlation coefficient r=−0.377, P=0.002), particularly in the initial stages of the disease (stages I and II; r=−0.368, P=0.020 and r=−0.480, P=0.002, respectively). Furthermore, this correlation was observed in the proliferative (r=−0.366, P=0.047) and the secretory phases (r=−0.321, P=0.049) of the menstrual cycle.  CONCLUSIONS: These results suggest that the reduced capability of endometrial tissue to downregulate IL‐1 proinflammatory effects may be involved in the increased expression of MCP‐1 in the endometrium of women with endometriosis and the establishment of an inflammatory state. The results also indicate a sustained process of cell activation throughout the menstrual cycle.
Analysis of the CD4 Protein on Human Vaginal T Lymphocytes
Authors: FIDEL PAUL L. , WORMLEY JR. FLOYD L. , CHAIBAN JOSEPH , LOUNEV VITALI , FIDEL PAUL L. , CHESSON RALPH R.
Published: December 2003

PROBLEM: Although T lymphocytes at the human vaginal mucosa have been partially characterized, there remains a paucity of information regarding cell‐mediated immune mechanisms at this mucosal site. In mice and humans, there are several phenotypic distinctions between vaginal T lymphocytes and those in the peripheral circulation. Recently, we observed as well that the N‐terminus of the CD4 protein on murine vaginal T lymphocytes is atypically expressed compared to its systemic counterpart, and that the atypical expression extends to the mRNA level.  METHOD OF STUDY: The purpose of this study was to evaluate the CD4 protein on human vaginal T lymphocytes by flow cytometry and RT‐PCR.  RESULTS: Results showed that, in contrast to mice, the CD4 protein on human vaginal and peripheral blood T lymphocytes are similar at both the molecular and protein levels.  CONCLUSIONS: These results indicate that based on several differences between human and mouse vaginal T cells, caution is urged when using mice as a model to study human vaginal immunity.
Pregnancy‐Specific Glycoproteins Function as Immunomodulators by Inducing Secretion of IL‐10, IL‐6 and TGF‐β1 by Human Monocytes
Authors: SNYDER SARA K. , WESSELLS JENNIFER L. , WATERHOUSE ROSEANN M. , DVEKSLER GABRIELA S. , WESSNER DAVID H. , WAHL LARRY M. , ZIMMERMANN WOLFGANG
Published: December 2003

PROBLEM: Low levels of pregnancy‐specific glycoproteins (PSGs) in maternal serum have been correlated with complications of pregnancy. We investigated the ability of human PSGs to regulate in vitro production of cytokines.  METHOD OF STUDY: Human monocytes and murine RAW 264.7 cells were treated with recombinant PSG1, PSG6, PSG11, or a truncated PSG6 consisting of only the N‐terminal domain (PSG6N). Cytokine production in response to PSG‐treatment was measured by ELISA and/or reverse transcriptase‐PCR.  RESULTS: All PSGs tested induced secretion of interleukin (IL)‐10, IL‐6 and transforming growth factor (TGF)‐β1 by both human and murine cells, but not IL‐1β, tumor necrosis factor (TNF)‐α or IL‐12. The N‐terminal domain of PSG6 was sufficient for induction of monocyte cytokine secretion. Induction of IL‐10 and IL‐6 was preceded by an increase in the specific mRNAs.  CONCLUSIONS: PSG1, PSG6, PSG6N, and PSG11 induce dose‐dependent secretion of anti‐inflammatory cytokines by human monocytes. Human and murine PSGs exhibit cross‐species activity. Our results are consistent with a role for PSGs in modulation of the innate immune system.
Plasma Antiviral Activity and Interferon‐γ Production by Superantigen‐stimulated Lymphocytes during Normal Human Pregnancy
Authors: GRASSO GIOVANNI , MASSAI LAURETTA , MIGLIACCIO PIERLUIGI , PICCIOLINI ENRICO , MUSCETTOLA MICHELA
Published: December 2003

PROBLEM: Plasma interferon (IFN)‐γ levels, lymphocyte responsiveness, and evaluation of the relationship between circulating antiviral activity (AA) and IFN‐γ production were studied in pregnant women and nonpregnant age‐matched controls with the objective of elucidating the downregulation of IFN‐γ production in successful pregnancy.

METHOD OF STUDY: In plasma and supernatants of peripheral blood mononuclear cell (PBMC) cultures, stimulated with staphylococcal enterotoxin A (SEA) superantigen, from 43 pregnant women with a history of normal pregnancy and 30 healthy nonpregnant age‐matched controls, levels of AA were measured in a micromethod by inhibition of the cytopathic effect (CPE) caused by vesicular stomatitis virus (VSV) in the human amnionic cell line (WISH).

RESULTS: Significantly higher plasma AA (60% was IFN‐γ and residual activity was acid‐labile IFN‐like) was present in pregnant women than controls. On the other hand, SEA‐activated PBMCs from pregnant women produced significantly lower IFN‐γ levels than those of nonpregnant women. Furthermore, maternal plasma AA levels correlated negatively with IFN‐γ production by SEA‐stimulated PBMCs.

CONCLUSION: The hypothesis that successful pregnancy requires downregulation of IFN‐γ is only partially sustained, suggesting that the immunology of pregnancy is more complex and that murine and human pregnancy have different cytokine profiles.
Blocking Antibodies in Blood from Patients with Recurrent Spontaneous Abortion in Relation to Pregnancy Outcome and Intravenous Immunoglobulin Treatment
Authors: JABLONOWSKA BARBARA , KJELLBERG SVANTE , SELBING ANDERS , PALFI MIODRAG , ERNERUDH JAN
Published: December 2003

PROBLEM: To study whether the occurrence of mixed lymphocyte culture (MLC) blocking antibodies is associated with pregnancy outcome in women with unexplained recurrent spontaneous abortion (RSA) and the in vivo effect of intravenous immunoglobulin (IVIG) treatment on MLC blocking effect.  METHOD OF STUDY: Blood samples from 41 RSA patients were obtained before and after pregnancy, and blocking antibodies were estimated by one‐way MLC assay. The patients received IVIG or placebo (saline) during pregnancy. Additionally, pre‐pregnancy blood samples from 31 RSA women and 10 controls were obtained.  RESULTS: We found no correlation between blocking antibodies before pregnancy and the pregnancy outcome. The occurrence of blocking antibodies was not affected by pregnancy or IVIG treatment.  CONCLUSIONS: Blocking antibodies have no predictive value for the pregnancy outcome in RSA patients, and their production seems not to be affected by IVIG.
IgG Anti‐laminin‐1 Autoantibody and Recurrent Miscarriages
Authors: INAGAKI JUNKO , MATSUURA EIJI , KAIHARA KEIKO , KOBAYASHI KAZUKO , YASUDA TATSUJI , NOMIZU MOTOYOSHI , SUGIURA‐OGASAWARA MAYUMI , KATANO KINUE , AOKI KOJI
Published: December 2003

PROBLEM: The present study assesses the clinical significance of anti‐laminin‐1 auto‐antibodies (auto‐Abs) in recurrent miscarriages.  METHOD OF STUDY: A total of 207 recurrent aborters with a history of two or more consecutive first‐trimester miscarriages were tested for the presence of anti‐laminin‐1 Abs, β2‐glycoprotein I‐dependent anticardiolipin Abs, lupus anticoagulants, anti‐DNA Abs, and anti‐nuclear Abs, before they had conceived again. Recurrent aborters then were followed up during subsequent pregnancies and their outcomes were evaluated relative to their blood test results prior to pregnancy. 
RESULTS: Fifty‐five (31.1%) women out of 177 recurrent aborters were positive for IgG anti‐laminin‐1 auto‐Abs. The levels of IgG anti‐laminin‐1 auto‐Abs in recurrent aborters were significantly higher than those in healthy pregnant women and in healthy non‐pregnant women (P=0.0043 and 0.0073, respectively). The live birth rate of subsequent pregnancies in IgG anti‐laminin‐1 auto‐Abs‐positive recurrent aborters was significantly lower than the IgG anti‐laminin‐1 auto‐Abs‐negative recurrent aborters (P=0.0320). There were no specifically significant relationships observed between IgG anti‐laminin‐1 auto‐Abs and other tested auto‐Abs.  CONCLUSION: IgG anti‐laminin‐1 auto‐Abs are associated with recurrent miscarriages and the subsequent pregnancy outcome of recurrent aborters.
Involvement of Calpain for Apoptosis in Dysfunction of the Unaffected Testis in Rats with Experimental Testicular Torsion
Authors: UMEMOTO YUKIHIRO , SASAKI SHOICHI , TATSURA HIROYUKI , KUBOTA HIROKI , KUBOTA YASUE , KOHRI KENJIRO , OZAKI YASUHIKO , SASAKI MAKOTO , OZAKI YASUHIKO
Published: December 2003

PROBLEM: The dynamics of calpain and involvement of apoptosis in sperm formation disorder of the unaffected testis in rats with experimental testicular torsion were investigated.  METHODS OF STUDY: Using 6‐week‐old Wistar rats, an experimental unilateral testicular torsion model was prepared. The bilateral testes were excised 1, 3, 5, 7, 14, 35, and 70 days after the left testis was twisted, and the unaffected testes were subjected to immunohistological staining, sodium dodecyl sulfate polyacrilamide gel electrophoresis and Western blotting using anti‐calpain antibodies. Apoptosis was detected by the TdT‐mediated dUTP–biotin nick end‐labeling (TUNEL) method.  RESULTS: By immunohistological staining, positive immunostaining by anti‐pro μ‐calpain antibody was observed in the spermatocyte nucleus, but not with anti‐pro m‐calpain antibody. The staining was increased until 7 days after testicular torsion, then decreased with progression of sperm formation disorder. By Western blotting, the intensity of staining with anti‐pro μ‐calpain antibody was increased until 7 days after torsion of the testis. Apoptosis expression in the unaffected testis was significantly inhibited by addition of a calpain inhibitor.  CONCLUSIONS: It was suggested that μ‐calpain may be involved in apoptosis expression in sperm formation disorder of the unaffected testis in unilateral testicular torsion.
Image Analysis of the Effects of Gonadectomy on Colloidal Carbon uptake In Vivo by Sinusoidal Cells in Liver and Spleen of the Mouse
Authors: OOGA TAKUJI , ITOH MASAHIRO , MIYAMOTO KENSAKU , TAKEUCHI YOSHIKI , NISHIOKA MIKIO
Published: December 2003

PROBLEM: The presence of androgen and estrogen receptors in macrophages was recently described. This suggests that both sex hormones directly act on reticuloendothelial system (RES) activity. Studies of the effect of gonadectomy on uptake and clearance of foreign materials make clear a role of the testis and ovary in RES regulation. However, there is still some disagreement as to the gonadal influence on RES.  METHOD OF STUDY: We applied a computed image analysis system to the investigation of the effects of gonadectomy on colloidal carbon uptake by the liver and spleen sinusoidal cells in mice. Mice were gonadectomized or sham‐operated. After a few weeks, they received colloidal carbon with or without pretreatment with lipopolysaccharide (LPS). Thereafter, livers and spleens were taken for quantitative determination of the amounts of endocytosed carbon in the sinusoidal cells by means of digital image analysis.  RESULTS: Ovariectomy did not significantly affect hepatic and splenic RES activities compared with sham‐operated controls. Treatment of mice with LPS after ovariectomy also caused no significant changes of the RES activities compared with sham‐operated and LPS‐injected controls. In contrast, orchidectomy resulted in significant enhancement of splenic RES activity compared with sham‐operated males, while hepatic RES activity did not significantly change with orchidectomy alone. However, LPS treatment to orchidectomized mice significantly augmented both hepatic and splenic RES activities compared with sham‐operated and LPS‐injected controls.  CONCLUSION: The significant augmentation of RES activity by orchidectomy, but not by ovariectomy, indicates that male RES activity is physiologically downregulated in the presence of the testes. Furthermore, splenic RES is more sensitive to orchidectomy than hepatic RES.