bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Use of ELISA to Evaluate Human Antibody Binding to Epididymal Sperm From Different Species
Authors: FICHOROVA RAINA N. , NAKOV LYUDMIL S. 
Published: May 2013

PROBLEM: The impact of antibodies to epididymal sperm antigens in human infertility has been poorly understood. Cross‐reactivity of human antibodies with animal epididymal sperm has been previously observed, however, only by means of qualitative methods. Moreover, it has been always compared to reactivity against human ejaculated rather than human epididymal sperm.METHOD: Following a screening study of 940 infertility patients, sperm agglutinating and immobilizing sera as well as sperm anbitody negative controls were used to standardize an ELISA employing human ejaculated sperm. Nine sera positive in ELISA were further tested against epididymal human, guinea pig, rat, and hamster sperm. Differences among groups were evaluated by factorial analysis of variance (ANOVA).RESULTS: The specificity and sensitivity of ELISA were shown to be 85.1% and 61.18%, respectively. Eight out of nine antisperm antibody‐positive sera from infertile subjects reacted relatively stronger with epididymal than with ejaculated human sperm. All tested infertility sera showed strong although variable cross‐reactivity with sperm from guinea pig, hamster, and rat.CONCLUSION: ELISA has definite potential in sperm antibody research, allowing quantitative assessment of the results and immotile sperm employment. The suggested predominant role of epididymal sperm antigens in immune responses related to fertility needs further investigation. Some of these antigens are obviously phylogenetically perserved, and possibly in a quantitative aspect present differently on epididymal spermatozoa from various mammalian species.
The Effect of Aspirin on Recurrent Fetal Loss in Experimental Antiphospholipid Syndrome
Authors: KRAUSE ILAN , BLANK MIRI , GILBRUT BORIS , SHOENFELD YEHUDA 
Published: May 2013

PURPOSE: To evaluate the effect of aspirin treatment upon fetal loss in mice with experimental antiphospholipid syndrome (APLS).MATERIALS AND METHODS: Experimental APLS was induced in pregnant mice by passive transfer of mouse monoclonal anticardiolipin antibody. The mice were treated with high (100μg/d) or low (10μg/d) does of aspirin, using vitaminC(100μg/d or 10μg/d)as a control. The mice were assessed for the presence of lupus anticoagulants (prolonged aPTT), thrombocytopenia, degree of fetal resorption rate and mean embryo and placental weights.RESULTS: The mice with APLS had a higher fetal resorption rate(45.7± 12.2% vs 2.5 ± 0.4%, P<0.001), reduced placenta mean weight(104 ± 8 mgvs 169 ±7mg, P<0.001), prolonged aPTT (94± 14sec vs 39±4sec, P<0.001), and reduced mean platelet count(597± 186 ± 103/mm3vs 847±51 ± 103/mm3,P<0.001). The groupof mice with APLS, who were treated with low‐dose aspirin, had a lower resorption rate (11.1 ±9.3% vs 45.7±12.2%, P<0.001), a higher placenta mean weight (178 ± 8 mg vs 104 ± 8 mg, P<0.001), a higher mean embryo weight (1042 ± 134 mg vs 721±91 mg, P<0.001), and a lower aPTT (58±15 sec vs 94±14 sec, P, <0.001). Micewho were treated with high‐dose aspirin also had a lower resorption rate, although not as much as in the low‐dose aspirin group (34.2 ± 12.7% vs 45.7 ± 12.2%, P<0.001).CONCLUSION: Aspirin, especially in low dose, has a protective effect against obstetrical complications associated with experimental APLS.
Maternal‐Fetal Histocompatibility in Intrauterine Growth Retarded and Normal Weight Babies
Authors: VERP MARION S. , SIBUL MARK , BILLSTRAND CHRIS , BELEN GARY , HSU MICHAEL , OBER CAROLE 
Published: May 2013

PROBLEM: To determine whether risk for intrauterine growth retardation (IUGR) is increased in HLA‐DQA1 compatible pregnancies.METHOD: Paired maternal and cord blood samples were obtained from 30 IUGR and 31 non‐IUGR pregnancies delivered at a university hospital. Samples were typed for eight HLA‐DQA1 alleles using 10 sequence‐specific oligonucleotide probes. Associations between IUGR and HLA‐DQ compatibility status, and other risk factors were examined using logistic regression analysis.RESULTS: HLA‐DQ compatibility and history of spontaneous abortion were not individually significant risk factors for IUGR; however, there was an interactive effect between these two factors and IUGR (P = 0.085) that improved the overall fit of the logistic model (P < 0.001). No individual allele was more common in IUGR pregnancies.CONCLUSION: HLA compatibility per se is not associated with sufficient inhibition of fetal growth to result in IUGR as defined. However, in women with a history of spontaneous abortion, HLA compatibility may have an effect.
Murine Trophoblast Failure and Spontaneous Abortion
Authors: CLARK DAVID A. , BANWATT DALJEET , CROY B. ANNE 
Published: May 2013

PROBLEM: Infection has been proposed to initiate abortion, and the role of viruses in spontaneous resorption in mice has not been tested.METHOD: The anti‐viral drug ribavirin (1‐β‐D‐ribofuranosyl‐1,2,4‐triazole‐3‐carboxamide) was fed to CBA/J and C3H/HeJ female mice beginning on the morning after mating with DBA/2J males.RESULTS: Ribavirin treatment increased the rate of abortion (resorption) on day 13.5, and this was associated with retardation of the rate of embryo development and hypoplasia of the trophoblast. There was a reduction in trophoblast‐dependent decidua‐associated soluble suppressor activity, but there was no maternal mononuclear cell infiltrate of the type reported in association with resorption of semiallogeneic and xenogeneic mouse embryos. This may be due to an immunosuppressive effect of ribavirin. Ribavirin was able to potently suppress proliferation of mouse trophoblast and mastocytoma cell lines in vitro.CONCLUSIONS: There are several drug‐induced murine abortion models that provide useful insights into potential mechanisms underlying spontaneous pregnancy failure, but in the ribavirin model, a direct impairment of trophoblast development appears to be responsible.
In Vitro Effect of Anticardiolipin Autoantibodies Upon Total and Pulsatile Placental hCG Secretion During Early Pregnancy
Authors: SHURTZ‐SWIRSKI R. , INBAR O. , BLANK M. , COHEN J. , BAKIMER R. , BARNEA E.R. , SHOENFELD Y. 
Published: May 2013

BACKGROUND: The anticardiolipin syndrome is characterized among other features by recurrent thromboembolic events, thrombocytopenia, and recurrent fetal loss associated with high IgG titers of anticardiolipin antibodies and/or the presence of lupus anticoagulant. AIMS: The mechanisms for the fetal loss in this syndrome have not yet been clearly elucidated, although several hypotheses based on experimental data have been put forward. We wanted to evaluated the effect in vitro of anticardiolipin antibodies on the secretion of human chorionic gonadotropin.METHODS: Employing our previous experience with placental explants, we studied the effect of several mouse monoclonal and human polyclonal purified anticardiolipin antibodies (ACA), which were shown by us to induce experimental antiphospholipid syndrome (APLS), to affect the pulsatile secretion of beta human choriogonadotropin.RESULTS: The mouse monoclonal ACA antibodies caused an increase in the pulsatility of beta human choriogonadotropin, while human polyclonal ACA derived from patients with ACA had an inhibitory effect.CONCLUSION: These studies with placental explants show that ACA may have an additional effect on placental hormone secretion and thus affect the fate of the embryo.
Mode of Inheritance of HLA Haplotypes Locus A,B in Siblings of Different Sexes
Authors: KOMLOS L. , LIVNI E. , KLEIN T. , NOTMANN J. , LEVINSKY H. , HALBRECHT I. , HART J. , ZAIZOV R. 
Published: May 2013

METHOD: Forty‐eight parents and 172 children were typed for class I HLA antigens, locus A,B.RESULTS: Although the number of cases is small, we observed: (1) a significantly decreased number of sons born after a first delivery of a son, as compared to a first delivery of a daughter; (2) significantly increased sharing of maternal class I HLA antigens between the firstborn son and his brothers from higher birth orders, as compared to his sisters; and (3) HLA‐A2 antigen, which is known to be involved in HLA restricted cytotoxic reactions in the recognition of minor histocompatibility antigens, was inherited in subsequent deliveries of sons as compared to daughters in a significantly higher frequency from the paternal than from maternal HLA haplotype. The results suggest that sharing of identical maternal HLA haplotypes between brothers may aid to decrease the degree of maternal sensitization to fetal antigens, and lack of HLA‐2 antigen in maternal cells from sons as compared to daughters may avoid maternal HLA‐A2 restricted cytotoxic reactions toward the male fetus.
Assay for Recombinant and Native Human Intraacrosomal Antigen SP‐10
Authors: SHEN MINCHENG , WRIGHT RICHARD M. , CARTA GIORGIO , HERR JOHN C. 
Published: May 2013

PROBLEM: To develop a method to measure a recombinant sperm protein, SP‐10, during scale up and purification for a contraceptive vaccine formulation.METHOD: A quantitative assay method for the human intraacrosomal protein SP‐10 was developed utilizing the format of indirect capture enzyme‐linked immunosorbent assay (ELISA). A SP‐10 specific monoclonal antibody mAb, MHS‐10, was used as the capture antibody. Two recognition reagents, a rabbit polyclonal anti‐SP‐10 antisera (pAb) and a biotin‐labeled mAb, MHS‐10, were used as the recognition antibodies, respectively. A SP‐10 recombinant fusion protein consisting of 125 SP‐10 amino acids linked to glutathione transferase was used as a working SP‐10 standard. The coefficient of variance for the assay system using the rabbit pAb was in the range of 0.099 to 0.157, and for the assay system using the biotinylated mAb MHS‐10 was in the range of 0.081 to 0.084.RESULTS: Employing biotinylated MHS‐10 as the recognition antibody, it was found that the native SP‐10 molecule had more than one MHS‐10 epitope. The concentration of SP‐10 in a pool of human sperm extracts was found to be approximately 1% of the total proteins, assayed by both of the recognition antibody systems.CONCLUSIONS: The assay system described is useful to monitor the yield of recombinant SP‐10 during scale‐up production of the SP‐10 vaccine.
Transforming Growth Factor β as an Immunosuppressive Protein in Human Seminal Plasma
Authors: NOCERA MARYANN , CHU T. MING 
Published: May 2013

PROBLEM: Human seminal plasma is known to exhibit immunosuppressive activity in seminal plasma.PURPOSE: The purpose was to characterize immunosuppressive proteins in seminal plasma.METHOD: Gel filtration fractions of 100 to >440 kDa were identified that inhibited DNA synthesis and killing activity of interleukin‐2 stimulated lymphocytes.RESULTS: The fractions exhibiting immunosuppression also inhibited DNA synthesis in a mink lung cell bioassay commonly used to measure the activity for transforming growth factor β (TGF‐β). The negative growth activity was diminished by aTGF‐β neutralizing monoclonal antibody. TGF‐β was further detected in the active fractions by Western immunoblot.CONCLUSIONS: These results identified TGF‐β as an immunosuppressive protein in human seminal plasma and may provide insight into the role of immunosuppression played by seminal plasma, such as in reproduction and neoplasia.
Lack of Expression of HLA Antigens on Immature Germ Cells From Ejaculates With Antisperm Antibodies
Authors: CASTILLA JOSE A. , GIL TERESA , RODRIGUEZ FERNANDO , MOLINA JORGE , SAMANIEGO FRANCISCO , VERGARA FRANCISCO , HERRUZO ALFONSO J. 
Published: May 2013

PROBLEM: The lack of expression of HLA antigen on immature germ cells from ejaculates with antisperm antibodies has been reported.METHOD: The expression of human leukocyte antigens on immature germ cells from ejaculates with antisperm antibodies (ASA) was investigated by indirect immunofluorescence using a panel of monoclonal antibodies (MAb) and automated flow cytometry. Patients were divided into two groups: fertile (prevasectomic; N= 10), and ejaculates with ASA (10 samples with IgG and IgA ASA, and five semen samples with only IgG ASA). ASA were detected on sperm using the direct immunobead test. After centrifuging semen samples on a Ficoll‐Hypaque gradient, round cells obtained at the gradient interface were gated by a flow cytometer. The “immature germ cell window” was defined in terms of cellular volume and granularity.RESULTS: The percentage of gated round cells from semen samples that reacted with anti‐CD45 was always less than 5%, and with anti‐CD44 less than 3%. This lack of reactivity of gated round cells with MAb specific for leukocytes and epithelial cells suggests that they were immature germ cells. Immature germ cells were unreactive with W6/32 and anti‐β‐2‐microglobulin MAb, which suggests that these cells do not express HLA class I molecules. Similarly, no reactivity of the immature germ cells with the MAb that recognize HLA class II molecules was found. No significant differences were observed in the expression of HLA molecules on immature germ cells between the different semen samples studied: fertile, and ejaculates with ASA.CONCLUSION: The presence of ASA in ejaculate is not associated with abnormal HLA antigen expression on immature germ cells.
GnRH Agonist Induces Suppression of Lymphocyte Subpopulations in Secondary Lymphoid Tissues of Prepubertal Female Mice
Authors: RAO L.V. , CLEVELAND R.P. , ATAYA K.M. 
Published: May 2013

PROBLEM: Gonadotropin‐releasing hormone (GnRH) agonists are playing an increasing role in the medical management of a variety of diseases. Recent evidence also indicates GnRH immune system interactions.METHOD: The present study investigated the sequential changes in lymphocyte subpopulations in secondary lymphoid tissues of prepubertal female mice in vivo following Lupron depot administration. A direct two‐color immunofluorescence staining followed by flow cytometric analysis was employed.RESULTS: Following agonist administration, white blood cell counts decreased significantly with decreases in both granulocyte and lymphocyte counts. Blood T‐cell and B‐cell subsets were also reduced although B cells decreased more markedly. In the spleen, B cells were again reduced more than T cells. There was no selective loss of either CD4 or CD8 subpopulations at any time point, in both spleen and blood. There were no differences in the percentage of lymph node subsets except that B cells decreased in the second week.CONCLUSION: These data indicate that GnRH agonist alters specific lymphocyte subpopulations and, therefore, have the potential for affecting immune system function in vivo.
No Detection of Interferon‐γ Activity During Early Pregnancy in the Mouse
Authors: LA BONNARDIERE CLAUDE 
Published: May 2013

PROBLEM: Interferons (IFN) have been shown to be secreted by the trophectoderm of implanting embryos in different species, in particular ungulates. In the pig, a clear‐cut IFN‐γ production, the role of which is unknown, was found in the trophoblast at implantation. A murine counterpart to these IFNs has not yet been identified.METHOD: Two sets of experiments were conducted to test the presence of IFN‐γ in the mouse conceptus. First, day 4 blastocysts were collected from Swiss mice and their antiviral activity measured in a microassay using mouse L 929 cells and vesicular stomatitis virus (VSV) in the presence or absence of anti‐IFN‐γ antibodies. In a second set of experiments, uteri from Swiss mice on days 5.5 and 6.5 of pregnancy were flushed and the resulting fluids assayed in a specific and sensitive ELISA test.RESULTS: In the antiviral assay, no consistent IFN‐like activity was found. The viral challenge also revealed a high susceptibility of mouse blastocysts to VSV infection. By ELISA, all but two samples (N = 75), whether on 5.5 or 6.5, were found negative.CONCLUSION: We conclude that in this rodent species IFN‐γ is most probably not involved in early maternal‐fetal interactions.
Estrogen Induces Expression of Endometrial Epidermal Growth Factor Receptors Before Implantation
Authors: KLEINSTEIN J. , WESTERMANN W. , MENNENGA K. , GUSTMANN CH. , GIPS H. 
Published: May 2013

PROBLEM: Pharmacological doses of 17β‐estradiol increase epidermal growth factor binding to uterine membranes.METHODS: To determine whether physiological elevated E2 concentrations in the preimplantation phase of rabbits and preovulatory phase of women cause epidermal growth factor (EGF) receptor expression, we performed radioligand binding studies of endometrial membranes with [125I]‐EGF.RESULTS: Unmated rabbits revealed a binding maximum (Bmax) of 514 ± 197 fmol/mg protein for [125I]‐EGF. The Bmax values increased significantly to 1424 ± 430 fmol/mg on day I and to 2244 ± 224 fmol/mg on day 2 after mating. On day 3 the Bmax values decreased to 1409 ± 238 fmol/mg and on days 4 to 8 the binding maxima were not significantly different from the Bmax values of unmated rabbits. In ovarieetomized rabbits a significant increase of [125I]‐EGF‐binding could be mimicked 12 and 18 h after the intramuscular injection of estradiol (40 μg/kg). The estradiol‐induced increase of [125I]‐EGF Bmax values.was blocked by the parallel application of the antiestrogen tamoxifen. Progesterone (2 mg/kg) did not influence [125I]‐EGF‐binding to endometrial membranes. Membrane preparations of the human endometrium from the periovulatory phase bound significantly more [125I]‐EGF compared to endometrium of the early proliferative phase. The expression of EGF receptor mRNA was localized in the epithelial cells of the human endometrium by in situ hybridization technique.CONCLUSIONS: These results provide insight into the regulation of the autocrine and paracrine factor EGF in the endometrium during the preimplantation period.
Enhanced Expression of Resident Leukocyte Interferon γ mRNA in Endometriosis
Authors: KLEIN NANCY A. , PÉRGOLA GABRIELA M. , RAO‐TEKMAL RAJESHWAR , DEY TAMMY D. , SCHENKEN ROBERT S. 
Published: May 2013

PROBLEM: Previous studies have shown that the endometrial epithelial/stromal cell proliferative activity of endometriosis is significantly less than that of normal endometrium and that the concentration of resident stromal leukocytes is significantly greater in ectopic than in eutopic endometrium. Other work has shown that interferon γ (IFNγ), secreted by resident leukocytes, inhibits endometrial cell proliferation in vitro. Accordingly, we hypothesized that the lower proliferative activity of endometriosis may be related to enhanced resident leukocyte IFN‐y production. This study was designed to assess whether resident leukocytes in endometriosis express IFNγ mRNA and to compare this expression to that of normal endometrium.METHODS: Biopsies of ectopic endometrium (N = 16) from women in the follicular phase and normal proliferative (N = 9) and secretory (N = 8) endometria were examined for IFNγ expression. Using monoclonal antibodies specific for CD45 (leukocyte common antigen), CD3 (a T‐cell marker) and CDI lc (a macrophage marker), leukocyte types were identified immunocytochemically, followed by in situ hybridization to examine expression of IFNγ mRNA.RESULTS: Results demonstrated that (1) the overall concentration of T cells and macrophages expressing IFN‐γ mRNA is significantly greater in endometriosis as compared to eutopic endometrium, and (2) the percent of each leukocyte type expressing IFNγ mRNA is greater in endometriosis than in normal endometrium.CONCLUSIONS: These findings support a possible paracrine role for resident leukocytes in regulating cell proliferation in endometriosis.
Human Embryo Associated Immunosuppressor Factor and Serum Estradiol Synergism in Ovarian Follicle Development
Authors: BOSE RATNA , KREDENTSER J. V. , MCCOSHEN JOHN A. 
Published: May 2013

PROBLEM: The purpose of this study was to test whether the level of serum immunosuppressor factor (EASF) correlates with ovarian follicle development and with serum hormones during the ovarian cycle.METHOD: One hundred seventy‐three serum samples were collected starting at the time of ovulation and 1 wk after insemination from 29 women attempting pregnancy by ovarian hyperstimulation and intrauterine insemination. After ovarian stimulation, the patients were given human chorionic gonadotropin (hCG) 38 h before insemination. Number and size of follicles were measured by ultrasound. Serum samples were tested for hCG. estradiol (E2). luteinizing hormone, follicle‐stimulating hormone (FSH), and progesterone levels. The level of EASF in patients' sera was assayed using a two‐site sandwich enzyme‐linked immunosorbent assay system. The levels of different hormones and EASF were then correlated among themselves and with the number of ovarian follicles.RESULTS: Eleven out of 29 patients had detectable levels of serum EASF‐Patients positive for EASF had a significantly higher number ovarian follicles ≥ 13 mm and E2 levels after hCG treatment as compared to patients with no detectable EASF. The levels of EASF correlated with number of follicles (P = .001). blood E2 (P < .02) and FSH levels (P < .02). Number of follicles ≥ 13 mm correlated with E2 (P = .01) and FSH levels (P < .01) only in patients positive for EASF.CONCLUSIONS: The levels of EASF, E2, and FSH may act synergistically in the development of ovarian follicles.
Mapping of Immunogenic Domains on Porcine Zona Pellucida 3 α and β Glycoproteins by Murine Monoclonal Antibodies
Authors: GUPTA S.K. , BAGAVANT HARINI , CHADHA K. , GUPTA M. , YUREWICZ E.C. , SACCO A.G. 
Published: May 2013

PROBLEM: Immunization with zona pellucida (ZP) leads to block in fertility with variable degree of ovarian dysfunctions. To design an immunocontraceptive vaccine based on synthetic peptides of zona pellucida, it is imperative to identify and define epitopes involved in sperm binding.METHOD: Epitopic domains recognized by monoclonal antibodies (MAbs) specific to either porcine ZP3α or ZP3β glycoproteins were delineated in an enzyme‐linked immunosorbent assay (ELISA) based on the ability of a MAb in solution to inhibit the binding of biotinylated ZP3 to another MAb coated on a microtitration plate. Immunoblot studies were carried out to determine the nature of reactive determinants. Porcine oocytes preincubated with MAbs were tested for sperm binding in vitro.RESULTS: Out of 23 MAbs generated, 10 had specificity for ZP3α and 13 for ZP3β. By using these antibodies, eight epitopic domains on both ZP3α and ZP3β were discernible. On ZP3β, epitopic domain DI partially overlaps with DII and DV with DVI, whereas on ZP3α domains DI to DV were in close proximity with a partial overlap, suggesting the dominance of this region. All 10 MAbs against ZP3α, and 10 out of 13 against ZP3β recognized deglycosylated forms of antigens. Seven antibodies having specificities for ZP3α and ZP3β respectively recognized linear epitopes. MA‐30, having specificity for ZP3β and MA‐420 for ZP3α and recognizing linear epitopes significantly inhibit the binding of boar sperm to porcine oocytes in vitro.CONCLUSIONS: Collectively, these studies indicate the value of utilizing MAbs for identifying and characterizing functionally significant ZP determinants. MAbs recognizing sequential epitopes will help in the elucidation of the amino acid sequence of the epitopes, which will subsequently help in design of synthetic immunocontraceptive vaccines.