bluuebunny/crossref_metadata_2025 huggingface dataset:
Three‐phase general border detection method for dermoscopy images using non‐uniform illumination correction
Authors: Norton Kerri‐Ann , Iyatomi Hitoshi , Celebi M. Emre , Ishizaki Sumiko , Sawada Mizuki , Suzaki Reiko , Kobayashi Ken , Tanaka Masaru , Ogawa Koichi
Published: September 2011

Background

Computer‐aided diagnosis of dermoscopy images has shown great promise in developing a quantitative, objective way of classifying skin lesions. An important step in the classification process is lesion segmentation. Many studies have been successful in segmenting melanocytic skin lesions (MSLs), but few have focused on non‐melanocytic skin lesions (NoMSLs), as the wide variety of lesions makes accurate segmentation difficult.

Methods

We developed an automatic segmentation program for detecting borders of skin lesions in dermoscopy images. The method consists of a pre‐processing phase, general lesion segmentation phase, including illumination correction, and bright region segmentation phase.

Results

We tested our method on a set of 107 NoMSLs and a set of 319 MSLs. Our method achieved precision/recall scores of 84.5% and 88.5% for NoMSLs, and 93.9% and 93.8% for MSLs, in comparison with manual extractions from four or five dermatologists.

Conclusion

The accuracy of our method was competitive or better than five recently published methods. Our new method is the first method for detecting borders of both non‐melanocytic and melanocytic skin lesions.
Flow‐Cytometric Characterization of Hematopoietic Cells in Non‐Pregnant Human Endometrium
Authors: Lachapelle M.‐H. , Miron P. , Hemmings R. , Baron C. , Roy D. C.
Published: September 2011

Lachapelle M‐H, Miron P, Hemmings R, Baron C, Roy DC. Flow‐cytometric characterization of hematopoietic cells in non‐pregnant human endometrium. Am J Reprod Immunol 1996; 35:5–13 cP Munksgaard, Copenhagen

PROBLEM: Immunologic evaluation and quantitation of hematopoietic cells in human endometrium has been difficult to perform, particularly in nonpregnant subjects. In this study, we describe a method for the flow‐cytometric characterization of hematopoietic cells present in the endometrium of non‐pregnant women.

METHOD: Endometrial biopsy samples from normal donors were first mechanically disrupted and filtered to generate a single‐cell suspension of leukocyte‐enriched endometrial cells. Cells were labeled with a panel of monoclonal antibodies, stained with propidium iodide (PI), and one‐ or two‐color flow‐cytometric analysis performed on cells excluding PI.

RESULTS: The methodology described in this study was highly reproducible in experiments evaluating the interrun and intrarun variability. We then determined the immunophenotypic profile of endometrial leukocytes from 12 normal females. The majority of leukocytes were T cells (CD3: 47%; CD4: 24%; CD8: 28%) with an important contingent of NK cells (CD56: 32%), the majority of which harbored the unusual CD16‐CD56 bright phenotype, and a minority of B cells (CD20: 6%) and monocytes (CD14: 7%).

CONCLUSIONS: Flow cytometry can be used to assess antigen expression on the surface of endometrial leukocytes from nonpregnant women. In future studies, it will be possible to use this approach to investigate the role of immune cell populations in the endometrium of patient experiencing reproductive failure.
Immunological Prevention of Spontaneous Early Embryo Resorption Is Mediated by Non‐Specific Immunosimulation
Authors: Baines Malcolm G. , Duclos Alain J. , de Fougerolles Antonin R. , Gendron Robert L.
Published: September 2011

PROBLEM: Spontaneous early embryo resorption following implantation occurs in many species, but little is known regarding the causes or the prevention of early pregnancy failure. Embryo and fetal loss have widely been assumed to be due to maternal allospecific immune rejection. Alloimmunization therapy with paternal tissues has been successfully used in human and murine pregnancies to prevent early embryo demise. The mechanisms of this treatment have been assumed to be the induction of antigen specific, fetal “graft” enhancing antibodies and suppressor cells. The purpose of this study was to investigate the validity of this assumption.

METHOD: To investigate these general assumptions, female CBA/J mice were immunized with either specific or nonspecific antigens prior to mating with DBA/2 or Balb/c males. Further, a model system for the study of bacterial lipopolysaccharide (LPS) induced abortion was used to demonstrate the nature of antigen specific immune protection against abortion.

RESULTS: Whereas the administration of 1 μg of LPS to CFW♀ x CFW♂ pregnant mice on day 7 of gestation induced complete fetal resorption, prior immunization with 20 μg of LPS completely prevented LPS induced abortion as long as the anti‐LPS antibody titers remained above a threshold value of about 1/500. Therefore, early embryo loss could be induced by a bacterial infection and could be prevented by appropriate immunity to abortogenic factors. However, due to the short half‐life of IgM antibodies, immunity to LPS was short‐lived and the protective effect of LPS immunization against LPS induced abortion waned after 5 wk. Through the use of the CBA/J♀ x DBA/2 ♂ model system to study spontaneous early embryo loss, previous vaccination of CBA/J female mice with Balb/c spleen cells expressing paternal MHC antigens, complete Freund's adjuvant (CFA) or LPS, all decreased the incidence of spontaneous resorption in subsequent pregnancies. Similarly, a previous mating with a Balb/c ♂ prevented spontaneous embryo loss for a period of up to 6 wk. However, none of the immunotherapeutic vaccinations or matings had a permanent effect on CBA/J ♀ × DBA/2 ♂ embryo survival, which one would have expected if specific immune mediators were involved.

CONCLUSION: The results of this study indicated that the decrease in the incidence of spontaneous embryo resorption following alloimmunization was more likely to be due to nonspecific immunomodulatory effects on the immune system of the female mice, as opposed to specific antipaternal immunity. This may, in part, explain the placebo effects observed for alloimmunization therapy for human habitual pregnancy loss. The relevance of these results to the development of immunotherapy strategies for the prevention of habitual abortion is discussed.
Influence of Histocompatibility Antigens in Recurrent Spontaneous Abortion Couples and on Their Reproductive Performances
Authors: Sbracia Marco , Mastrone Marcello , Scarpellini Fabio , Grasso John A.
Published: September 2011

PROBLEM: To determine if human leukocyte antigens (HLA) play any role in the aetiology of recurrent spontaneous abortion (RSA), a substantial group of RSA couples were studied, and their reproductive performances in a 3‐year follow‐up recorded.

METHODS: HLA typing was performed for HLA‐A, ‐B, and DR antigens in both partners of 75 couples with unexplained RSA, and compared with a control group of 30 fertile couples that never experienced abortion. A further 57 couples of this group were studied for their reproductive performance in a 3‐year follow‐up, and subdivided into three subgroups: 1) couples that achieved successful pregnancy during the follow‐up; 2) couples that experienced abortion and no livebirth during the follow‐up; and 3) couples that experienced infertility during the follow‐up.

RESULTS: There were no significant differences for antigen frequency in all the different HLA loci, and HLA antigen sharing between all the RSA couples and controls. Significant increase of sharing for HLA‐DR locus was observed in the couples that aborted during the follow‐up with respect to the couples that achieved livebirth and controls (P < 0.03 and P < 0.02 respectively), and significantly increased frequency of B44, DR5 antigen combination in the same comparison (P < 0.03). No significant differences were observed in terms of the interval between conceptions in couples without antigen sharing with respect to couples with 1, 2 or more antigens shared, and antigen sharing in Locus A, B or DR.

CONCLUSIONS: The results suggest that gene(s) disadvantageous for reproduction may exist between the HLA‐B and ‐DR chromosomal region which influences the pregnancy outcome in RSA couples, and that HLA‐antigen sharing itself does not influence the outcome.
ZP3 Peptide Vaccine That Induces Antibody and Reversible Infertility Without Autoimmune Oophoritis
Authors: Tung Kenneth S.K. , Ang Jennifer , Lou Yahuan
Published: September 2011

Autoantibodies to ZP3, a major glycoprotein of the zona pel lucida (ZP) with sperm receptor function, can block sperm/oocyte interaction. However, only mice of certain major histocompatibility complex (MHC) haplotype respond to the ZP3 peptide. Moreover, ZP3‐specific T cells can mediate ovarian autoimmune disease.

A chimeric peptide has been designed that induces antibody to native ZP3 regardless of the MHC haplotype of the inbred mice tested. This results in reduction in fertility that is reversible. Infertility correlates well with ZP antibody titer, and the mice do not develop concomitant autoimmune oophoritis. The vaccine contains (1) a promiscuous foreign T‐cell peptide capable of eliciting a T‐cell response regardless of the animals' MHC haplotype, and (2) a modified native B‐cell peptide of ZP3.
Immune Modulators in the Context of the Ovulatory Process: A Role for Interleukin‐1
Authors: Adashi E.Y.
Published: September 2011

Recent studies have established the existence of a complete intraovarian interleukin‐1 system replete with ligends, receptors, and a receptor antagonist. This article examines the hypothesis that intraovarian IL‐1 may play an intermediary role in the ovulatory process. More specifically, it is hypothesized that intraovarian IL‐1β may constitute a mediator of gonadotropins in the induction of ovulation, an effector capable of coordinating and amplifying key components of the ovulatory cascade. The insight derived from this investigation may result the elucidation of some of the molecular events underlying the ovulatory process, in the delineation of the reproductive side effects of systemic anti‐IL‐1 therapy, and in potentially improved strategies for the promotion of fertility or its control.
HLA Antigen Sharing in Italian Couples in Which Women Were Affected by Gestational Trophoblastic Tumors
Authors: Sbracia M. , Scarpellini F. , Mastrone M. , Grasso J.A.
Published: September 2011

PROBLEM: The development of gestational trophoblastic tumors (GTT), in which genetic factors are strongly involved, is a rare event. To test the possibility that gene(s) linked to the Major histocompatibility Complex (MHC) may have a role in both embryo growth and tumor development, the HLA typing was performed on patients affected by GTT and on their partners.

METHOD: The study group of sixteen couples, in which the women were affected by an invasive mole or choriocarcinoma, and the control group of thirty normal fertile couples without history of spontaneous abortion or GTT were typed for class I and class II HLA antigen.

RESULTS: The results showed no differences in single HLA‐A and B antigen frequency between GTT couples and controls. In HLA‐DR, locus an increased frequency of DR‐6 antigen was observed (p<0.05). No differences were observed in the frequency of number of antigens shared. When considering the single locus no differences were found in the sharing of the antigens of the A and B locus, while the frequency of antigenic sharing for DR locus was significantly higher in GTT couples with respect to controls (p<0.025). Furthermore a higher frequency of Bw35‐DR5 antigenic combination was found in GTT partners than in controls (P<0.02).

CONCLUSIONS: These data represent a confirmation of the existence of a MHC linked gene(s) influencing the GTT development.
MUC1 as a Cell Surface and Secretory Component of Endometrial Epithelium: Reduced Levels in Recurrent Miscarriage
Authors: Aplin J.D. , Hey N.A. , Li T.C.
Published: September 2011

The mucin MUC1 is a large, highly glycosylated, hormonally regulated product of endometrial glandular and luminal epithelium with both cell surface‐associated and secreted isoforms. The abundance of mRNA coding for MUC1 increases about sixfold from the proliferative to the early secretory phase (Hey et al., J. Clin. Endocrinol. Metab. 78:337–342, 1994). Immunohistochemical studies show intracellular deposits accumulating in the early secretory phase followed by the release of MUC1 into gland lumens. The apical surface of luminal epithelium is strongly immunopositive in the early secretory phase. We have used a two site ELISA to measure MUC1 in uterine flushings as a function of time after the luteinising hormone (LH) peak. Low levels of secretory MUC1 are observed before day LH+7, while values on days LH+7‐LH+13 are much higher. Using semi‐quantitative immunohistochemical methods we have shown that in women suffering recurrent spontaneous miscarriage, mid secretory phase levels of MUC1 core protein and mucin‐associated glycans are reduced (Serie et al., Fertil. Steril. 62:989–996, 1994). Similarly, lower core protein levels are observed in uterine flushings after day LH+7 in these women. Reduced epithelial secretory function and a resultant change in uterine fluid composition are features of endometrium from recurrent miscarriage patients.
Development and Validation of an Assay for Measuring Preimplantation Factor (PIF) of Embryonal Origin
Authors: Roussev Roumen G. , Coulam Carolyn B. , Barnea Eytan R.
Published: September 2011

PROBLEM: Tests to determine presence of embryos prior to implantation are needed. METHODS: Sera from women after embryo transfer were tested for preimplantation factor (PIF) using the lymphocyte/platelet binding assay. Autorosettes were counted using blood type O+ donor lymphocytes and platelets incubated with blinded serum in the presence of antiCD2 antibody and rabbit complement. Human chorion gonadotropin (hCG) concentrations were determined 7 days later and compared with results of the lymphocyte/platelet assay. Implantation was confirmed by ultrasonographic evidence of presence of an intrauterine gestational sac. The roles of platelet activating factor (PAF) and chaperonin 10 in the observed phenomena were studied experimentally.

RESULTS: Significantly more lymphocyte/platelet rosette formations were observed when sera from women who successfully implanted were compared to sera from women who failed to implant. Neither PAF nor chaparonin added to the tested sera controls influenced the percentage of lymphocyte/platelets rosettes.

CONCLUSIONS: PIF is a likely candidate to be the next frontier of diagnosing the presence of viable preimplantation embryos in vivo.
Interleukins‐6 and −8 Levels in Sera and Cervical Mucus of Fertile, Idiopathic Infertile, and Immunoinfertile Women: Implication in Infertility
Authors: Naz Rajesh K. , Butler Avigdor
Published: September 2011

PROBLEM: The purpose of this study was to investigate the levels of interleukin‐6 and −8 (IL‐6 and IL‐8) in sera and cervical mucus of infertile (idiopathic and immunoinfertile) women and to compare the levels with those in age‐matched normally cycling fertile women.

METHODS: Levels of IL‐6 and IL‐8 were measured in the sera and cervical mucus of fertile and infertile women by the enzyme‐linked immunosorbent assay (ELISA). A non‐parametric Mann‐Whitney test was used to evaluate significance between the means. Linear regression analysis of IL‐6 and IL‐8 concentrations in serum versus cervical mucus and with antisperm antibody titers was performed by condition as well as all groups together. P < 0.05 was considered significant.

RESULTS: Both IL‐6 and IL‐8 were detected in sera and cervical mucus of fertile as well as infertile groups. Although serum levels of IL‐6 and IL‐8 were significantly different between fertile and infertile groups, the differences were more pronounced in the cervical mucus samples. Cervical mucus of idiopathic and immunoinfertile women demonstrated significantly (P = 0.002 to < 0.0001) greater concentrations of IL‐6 and IL‐8 compared to fertile controls. In general, there was no significant correlation between the serum and cervical mucus IL‐6 and IL‐8 levels, whether analyzed by condition or all groups together.

CONCLUSIONS: Elevated levels of IL‐6 and IL‐8 in cervical mucus of infertile groups may play a role in etiology of idiopathic and immunologic infertility. These findings suggest that the measurements of cytokines (especially IL‐6 and IL‐8) in cervical mucus may provide a tool for specific diagnosis and possibly treatment of infertility, especially idiopathic infertility.
On the Use of Paramagnetic Beads and Ferrofluids to Assess and Eliminate the Leukocytic Contribution to Oxygen Radical Generation by Human Sperm Suspensions
Authors: Aitken R.J. , Buckingham D.W. , West K. , Brindle J.
Published: September 2011

PROBLEM: To develop a methodology to determine a) the leukocytic contribution to reactive oxygen species generation by human sperm suspensions and b) the therapeutic value of removing these cellular contaminants.

METHODS: Leukocytes were removed with paramagnetic beads or colloidal ferrofluids coated with anti‐CD45 antibody. The sperm suspensions were monitored for oxidant generation by chemiluminescence, leukocyte contamination by immunocytochemistry, and fertilizing potential using zona‐free hamster oocytes.

RESULTS: Percoll®‐prepared human sperm suspensions exhibited a competence for PMA‐induced reactive oxygen species generation which was significantly correlated with leukocyte contamination. However, the purified spermatozoa remaining after paramagnetic bead treatment, also demonstrated an intrinsic capacity for PMA‐responsive reactive oxygen species generation and, freed from the oxidative stress created by the leukocytes, exhibited a significantly enhanced capacity for sperm‐oocyte fusion.

CONCLUSIONS: Although human spermatozoa can generate reactive oxygen species, sperm function is inhibited by the additional oxidative stress created by contaminating leukocytes. Removal of these cells with paramagnetic beads enhances fertilizing potential.
Enhancement of the Expression of Progesterone Receptor on Progesterone‐Treated Lymphocytes After Immunotherapy in Unexplained Recurrent Spontaneous Abortion
Authors: Chiu Lin , Nishimura Motoko , Ishii Yasuo , Nieda Mie , Maeshima Masaki , Takedani Yuji , Shibata Yoichi , Tadokoro Kenji , Juji Takeo
Published: September 2011

PROBLEM: The immunological mechanism of an effective immunotherapy with paternal lymphocytes for unexplained recurrent spontaneous abortion (RSA) is not yet clear. Previous studies revealed that progesterone plays an important role in maintaining normal pregnancy and lower expression of progesterone receptor (PGR) on lymphocytes was found in RSA. Therefore, it was of interest to investigate whether immunotherapy for RSA would be able to enhance the expression of PGR on lymphocytes of RSA.

METHOD: PGR expression on lymphocytes was analyzed with indirect immunofluorescence using flow cytometry.

RESULTS: There was no change of PGR expression on PBL of RSA between pre‐ and post‐immunotherapy (P > 0.05), while in the presence of 10.0 ug/ml progesterone for 24 h, PGR expressed on PBL on post‐immunotherapy was increased significantly as compared with that of pre‐immunotherapy in successful cases (P < 0.05) and decreased in abortive cases (P < 0.05). Most PGR was expressed on both CD4+ and CD8+ lymphocyte subsets. In successful cases, CD8+PGR+ subset of post‐immunotherapy was found to be increased significantly (P < 0.05) in comparison with that of pre‐immunotherapy.

CONCLUSION: The data in the present study suggest that immunotherapy for RSA induced a higher expression of PGR on progesterone‐treated lymphocytes, which may be involved in successful pregnancy.
Immunoactive Products of Placenta. V. Immunoregulatory Properties of a Low Molecular Weight Compound Obtained From Human Placental Cultures
Authors: Djian Valentine , Menu Elisabeth , Thibault Gilles , Ropert Sylvie , Chaouat Gerard
Published: September 2011

PROBLEM: We have previously shown that supernatants from short‐term cultures of human placental explants (HPS) are immunosuppressive in vitro as well as in vivo. They contain a low M.W. factor endowed with immunoregulatories activities (Filtrate of such with a 5 kDa cut off). In this paper, we wanted to assess whether this low M.W. material accounts for most, if not all, of the immunosuppressive properties of crude HPS and begin to investigate its mode of action.

RESULTS: The filtrate is active across species barrier and inhibits human and murine PHA driven lymphocyte proliferation, Mixed Lymphocyte Reaction, and Natural Killer activity as did crude HPS. It does not affect CTL lytic function at effector stage. Its cross species activity allowed us to study its effects in vivo. It corrects resorbtions in the CBA x DBA/2 murine spontaneous abortion model, and suppresses local and general GVH reactions in a model (A cells into irradiated A x B F1s) relevant to a clinical use, e.g., bone marrow transplantation.

To ensure that such survival of the recipients was due to donor cells in the latter, surviving experimental animals were analysed by FACS for repopulating lymphocytes phenotype, which was indeed of donor origin.

To elucidate the mechanism(s) of action of the active HPS moiety, we first tested various malignant cell lines for the minimal incubation time required for maximal lymphocyte inhibition. In the same vein, we verified that lymphocytes stimulated by PHA and simultaneously treated with filtrate were unresponsive to a second PHA challenge. The effects of the material was reversible if cells were washed out of it early enough before otherwise entering a cycle leading ultimately to cell death in vitro. Finally, we tested several second messenger pathways, none of which were modified.

CONCLUSION: These data suggest that the filtrate contains an entity that represents the main, if not all, the immunosuppressive molecules present in HPS. In addition, they suggest that the material acts only on activated T cells and requires to be present early in the replication activation cycle.

Altogether, the in vitro data strongly suggest that the material is acting by inducing clonal deletion in activated (T) cells.
Human Trophoblast Cell Adhesion to Extracellular Matrix Protein, Entactin
Authors: Yang Yan , Todt Jill C. , Svinarich David M. , Qureshi Faisal , Jacques Suzanne M. , Graham Charles H. , Chung Albert E. , Gonik Bernard , Yelian Frank D.
Published: September 2011

PROBLEM: Trophoblast interaction with endometrial extracellular matrix (ECM) is crucial during human embryo implantation and placentation. Entactin, a ubiquitous basement membrane glycoprotein, plays a central role in ECM assembly, cell attachment, and chemotaxis. The present study was conducted to examine the possible role of entactin in promoting human trophoblast adhesion.

METHODS: Using an extended life span first trimester trophoblast cell line HTR‐8/SVneo (HTR) and a cell adhesion assay, we measured the adherence of human first trimester trophoblasts to recombinant entactin and its domains. Also, we used flow cytometry and indirect immunofluorescence to detect the presence of integrins that may be involved in human trophoblast‐entactin interaction; these methods were used to analyze HTR cells, as well as tissue sections and freshly isolated human trophoblasts from first trimester and term placenta.

RESULTS: We found that first trimester trophoblast cells were highly adherent to entactin and its E and G2 domains but not to G1 or G3 domains. Using indirect immunofluorescence and flow cytometry, we found that both β1 and β3 integrin subunits were expressed on the surface of HTR trophoblast cells adhering to entactin; in contrast, β2 and β4 integrin subunits were not detected. In addition, we found that αvβ3 was expressed on freshly isolated villous cytotrophoblasts and cytotrophoblast and syncytiotrophoblasts in tissue sections from term placenta. The β3 integrin subunit was expressed in cytotrophoblasts and syncytiotrophoblasts in villi of first trimester placental tissue sections.

CONCLUSION: Recombinant entactin promotes human trophoblast cell adhesion through both its E and G2 domains and these specific adhesive interactions may be mediated by β1 and/or β3 class integrins.
Maternal Antibodies Protect Immunoglobulin Deficient Neonatal Mice From Mouse Hepatitis Virus (MHV)‐Associated Wasting Syndrome
Authors: Gustafsson Erika , Blomqvist Gunilla , Bellman Anna , Holmdahl Rikard , Mattsson Anita , Mattsson Ragnar
Published: September 2011

PROBLEM: Neonatal mice nursed by dams lacking immunoglobulins (Igs) may often suffer from lethal runting if raised under conventional conditions. The present study was performed in order to clarify a) the cause of the wasting syndrome and b) the protective role of antigen‐specific milk antibodies.

METHOD: Ig‐deficient mouse embryos in a conventional environment were embryo‐transferred to specified pathogen free (SPF) dams. Neonatal growth, mortality, and health status of mice from both environments was recorded. Suspected presence of mouse hepatitis virus (MHV) was tested by RT‐PCR. Protective effects on neonatal mortality of milk containing different titers of anti‐MHV antibodies were investigated in cross‐fostering experiments.

RESULTS: The SPF colony of Ig‐deficient mice exhibited no breeding problems, whereas Ig‐deficient neonates in the conventional environment suffered from lethal wasting syndrome. Serological screening of the mice kept in the two environments revealed that mice in the conventional room had high titers of antibodies against mouse hepatitis virus. Presence of MHV in runting neonates was confirmed by pathological examinations and RT‐nested‐PCR using MHV genome specific primers. Milk containing high titers of anti‐MHV antibodies, when provided for 8 days or more, completely prevented Ig‐deficient neonates from developing wasting syndrome in the conventional environment.

CONCLUSION: These findings show that the neonatal wasting syndrome is associated with the presence of MHV and that neonates are efficiently protected by MHV‐specific antibodies in the milk.
Characterization of a Sperm Nuclear Protein
Authors: Batova Iglika N. , Petrov Malcho G. , Kyurkchiev Stanimir D. , Kehayov Ivan R.
Published: September 2011

PROBLEM: The molecular identity of sperm DNA‐binding structural proteins contributing to the integrity of a sperm residual high salt/nuclease resistant nuclear structure is studied by cDNA cloning and monoclonal antibodies to the recombinant polypeptide. This structure, which is likely to be transferred unimpaired in the oocyte and is anticipated as a molecular correlate of the nuclear scaffold (nuclear matrix/envelope) in somatic cells, may be essential with respect to its DNA organization for the recovery and assembly of somatic‐type chromatin in the zygote. Recently, a cDNA encoding one of these proteins has been cloned and the recombinant polypeptide expressed in E. coli as a β‐galactosidase fusion protein. The main objective of the present study is the identification of the native sperm antigen by monoclonal antibodies raised against the recombinant molecule.

METHODS: We evaluated the possibility of immunizing by direct intrasplenic deposition in BALB/c mice of the recombinant fusion protein available as transblotted on nitrocellulose membrane carriers or as nitrocellulose protein‐bearing particles. Isolated sperm DNA/tight binding protein complexes were used in ELISA and Western blotting for selection of monoclonal antibodies specific to self epitopes of the nuclear antigen, as well as immunofluorescence of swollen human spermatozoa subjected to in situ extraction with high salt/β‐mercaptoethanol/DNase I and proteolysis, and of a cultured fibroblast cell line L‐929.

RESULTS: A monoclonal antibody, Mab 2C4, was selected which recognized a 52 kDa protein in the fraction of sperm high salt/urea resistant proteins. The target polypeptide was detected on swollen spermatozoa primarily to the post‐acrosomal and/or equatorial regions whereas in nonextracted sperm cells the epitope was exceedingly unavailable. The somatic cell location of the cognate epitope was confined to the nuclear envelope displaying a caplike pattern of staining, and also in a juxtanuclear cytoplasmic randomly coiled filamentous network and in compact bodies.

CONCLUSIONS: A nuclear protein salt‐stably bound to the sperm residual structure has been identified. The antigen appears localized in sperm exclusively to perinuclear subacrosomal sites that may be anchored at the male nuclear envelope, given the occurrence of the target epitope in somatic cells as well in nuclear and cytoplasmic sites adjacent to the nuclear membrane.
Antiphospholipid Antibodies in Patients With Preeclampsia
Authors: Allen Joseph Y. , Tapia‐Santiago Cecile , Kutteh William H.
Published: September 2011

PROBLEM: To determine the incidence of anticardiolipin and antiphosphatidylserine antibodies in women with preeclampsia.

METHODS: Sera from 100 women with preeclampsia and 100 normotensive pregnant women in the third trimester were assayed for anticardiolipin and antiphosphatidylserine antibodies.

RESULTS: Antiphosphatidylserine antibodies were positive for IgM in 1 patient (1%) and for IgG in 4 patients (4%). IgM antibodies to cardiolipin were positive in two patients (2%) while IgG antibodies to cardiolipin were positive in nine patients (9%). Only 3% of the control women were positive for antiphospholipid antibodies. None of the patients or controls had positive levels of IgA anticardiolipin or antiphosphatidylserine antibodies.

CONCLUSIONS: Elevated levels of IgG or IgM antibodies to cardiolipin and phosphatidylserine were detected in 11/100 (11%) of women diagnosed with preeclampsia in the third trimester compared to only 3/100 (3%) positive in controls (P < 0.05). These findings suggest that antiphospholipid antibodies may play a pathogenic role in some women with preeclampsia.
Production of Embryotoxic IgG Antibodies During IFN‐γ Treatment of Pregnant Mice
Authors: Athanassakis Irene , Aifantis Yiannis , Ranella Anthi , Vassiliadis Simon
Published: September 2011

PROBLEM: Administration of IFN‐γ during pregnancy in mice is deleterious not only to fetal survival but also to maternal physiology. Thus, injection of recombinant IFN‐γ from days 6–11 of gestation results in significant increase of fetal abortion, decrease of fetal weight accompanied by morphological defects of the embryo, and induction of class II MHC antigens on the spongiotrophoblast zone of the placenta. At the maternal level, this treatment causes splenomegaly, decrease of hematocrit levels, and increase of IgG production. In an attempt to dissect out the different phenomena observed, we examined the properties of polyclonal IgG antibodies contained in the animals' serum as to their ability to recognize antigenic determinants on IFN‐γ‐induced placentae and isolated trophoblasts.

METHOD: Serum from IFN‐γ‐treated pregnant mice was tested in vitro for its ability to recognize specific structures on primary trophoblasts and placental sections induced by IFN‐γ. In vivo this serum was injected in pregnant mice, and the outcome of pregnancy was evaluated. Monoclonal antibodies, resulting from the fusion of spleen cells from IFN‐γ‐treated pregnant mice to a myeloma cell line, were used to certify the IgG‐dependent embryotoxic effects observed with the polyclonal serum.

RESULTS: It was demonstrated that both the polyclonal serum and the monoclonal antibodies recognize antigenic determinants only on the IFN‐γ‐induced trophoblasts, placentae, and embryos, reduce fetal size, and cause splenomegaly in the mother, but do not affect the percentage of abortions as compared to controls.

CONCLUSIONS: IFN‐γ induces specific protein(s) on trophoblasts, which are responsible for embryotoxic antibody production in the mother. Since human abortion has been correlated with the production of embryotoxic IgG antibodies, this animal model may prove to be a useful tool in the analysis of events leading to pregnancy loss.
Human Autoantibodies Recognizing Human and Mouse Preimplantation Stages
Authors: Nilsson B. Ove , Jin Meishan , Larsson Anders , Sundström Per
Published: September 2011

PROBLEM: To find out whether autoantibodies against human preimplantation stages are present in some human sera and, if so, whether the antibodies could be capable to affect the egg development and/or to trigger an activation of the complement system at the procedures of assisted conception.

METHODS: 1. Immunohistochemistry on blots of human preimplantation stages. 2. Immunohistochemistry on paraffin sections of human and mouse preimplantation stages. 3. Culture of mouse morulae to analyze complement activation.

RESULTS: 1. Some human sera contained autoantibodies against human preimplantation stages. 2. Human‐mouse cross‐reacting antibodies against preimplantation stages occurred. 3. Immune complexes, formed on mouse preimplantation stages, activated the complement systems in egg cultures, resulting in a damaging of the eggs.

CONCLUSION: The presence of natural autoantibodies to preimplantation stages may be associated with reproduction failure, caused by a direct effect by the autoantibodies and/or an activation of the uterine complement system by the immune complexes formed.
Effect of Immunization With Human SP‐10 in Male Rodents
Authors: Sehgal Dr. Shobha , Koul Dimpy , Verma Seema
Published: September 2011

PROBLEM: Over the years immunocontraception has emerged as a promising modality. Theoretically it is possible to intercept fertility using a panel of antigens expressed along the entire pituitary, hypothalamus and gonadal axis. One such sperm antigen designated as SP‐10 is in advanced stages of development. Its gene has been cloned by Wright et al. (Biol Reprod 1990; 42:693–701). It is envisaged that immunization with SP‐10 would induce anti‐sperm antibodies in females and hence interrupt fertility. However, the in vivo effect of SP‐10 immunization has been assessed in male rodents with particular reference to spermatogenesis and effect of such immunization in various tissues. The SP‐10 antigen was a generous gift from Dr. John Herr.

METHODS: A group of male Wistar rats and BALB/c mice were immunized with a total dose of 175 μg of SP‐10 glutathione transferase fusion protein. Anti‐SP‐10 antibodies generated were detected by ELISA, immunofluorescence, flow cytometry and western blot assay. Histopathology of all the organs was conducted with particular reference to assess their effect on sperm maturation in testicular sections.

RESULTS: All the animals immunized with SP‐10 depicted a significant antibody response. DNA analysis by flow cytometry did not reveal any arrest of spermatogenesis, which was confirmed by histological studies.

CONCLUSION: It is concluded that conventional immunization generating significant antibody titers does not induce arrest of spermatogenesis in male rodents.
Anti‐Seminal Plasma Antibodies Associated With Infertility: I. Serum Antibodies Against Normozoospermic Seminal Plasma in Patients With Unexplained Infertility
Authors: Fichorova R.N. , Boulanov I.D.
Published: September 2011

PROBLEM: To determine whether patients whose infertility remains unexplained (by routine physical and laboratory investigations including sperm antibody testing) exhibit significantly elevated humoral anti‐seminal plasma immune responses in comparison to fertile individuals.

METHOD: Sera from 72 infertile couples were examined by ELISA for the presence of antibodies against normozoospermic seminal plasma (NSP). A group of 36 fertile individuals was used to determine the level of physiological reactivity. Kibrick's and Friberg's sperm‐agglutination tests were employed to distinguish patients routinely diagnosed as immunologically infertile.

RESULTS: Anti‐NSP reactivity was elevated in 18% of male and 26% of female patients. In the female patients, the prevalence rate for elevated anti‐NSP reactivity was significantly higher among partners of positive men. In only 4% the positive anti‐NSP results coincided with sperm‐agglutinating antibodies.

CONCLUSION: NSP constituents other than sperm‐coating antigens might interfere with infertility related autoimmune imbalance. The identification of the corresponding NSP antigens might help to resolve more cases of unexplained infertility.
Use of Chemiluminescent Method for Evaluation of Postoperative Adhesion Formation After Reproductive Pelvic Surgery: An Experimental Study
Authors: Myinbayev Ospan A. , Rubliova Ksenia I.
Published: September 2011

Surgery on the uterine horns was performed on 160 animals with application of catgut (CG) and prolene (PN), as well as the assisting application of interceed (TC 7) and fibrin glue (FG). The data of chemiluminescence (CL) of peritoneal phagocytes (PP) were compared with the results of evaluation of reparative wound regeneration. It was determined that the initial level and spontaneous CL activity of phagocytes in healthy animals was low or there were none of them that testified to a quiet cellular state. The maximal activating effect on PP was produced by CG, less so by FG, even less by TC 7, and the least by PN.
Evolution of Maternofetal Transport of Immunoglobulins During Human Pregnancy
Authors: Malek Antoine , Sager Ruth , Kuhn Peter , Nicolaides Kypros H. , Schneider Henning
Published: September 2011

PROBLEM: We determined the evolution of the maternal‐fetal transport of immunoglobulins during human pregnancy.

METHOD: Paired blood samples were collected between 17–41 weeks of gestation (WG) by puncture of a peripheral maternal vein and by cordocentesis (17–36 WG, n=91) or directly at delivery (37–41 WG, n=16) from the umbilical vein. Additional maternal samples were collected from the same individual (n=16) at 10,20,30 WG, and at term. The concentration of IgG and its four subclasses and of IgA were determined in the sera using ELISA method.

RESULTS: The mean level of IgG and IgA in maternal sera at 9–16 WG was 13.72±2.53 g/L and 3.95±1.23 g/L, respectively. Both, IgG and IgA throughout pregnancy decreased to a level of 60–70% (37–41 WG) of the initial concentration in early pregnancy. The ratio of IgG1:IgG2 in the maternal circulation was 2–3 and remained constant throughout pregnancy (17–41 WG). IgG3 and IgG4 levels remained constant and together were less than 10% of total IgG. In the fetal circulation a continuous rise in the level of both IgG and IgA was observed between 17 and 41 WG. Fetal level of IgG at 17–22 WG was only 5–10% of the maternal level and at term exceeded the maternal level reaching a value of 11.98±2.18 g/L. IgG1 at 17–22 WG was 0.93±0.42 g/L, which is approximately three times higher than IgG2. IgG, showed an exponential rise and at 37–41 WG its concentration was seven times higher than IgG2. IgG3 and IgG4 also showed an exponential rise and at term reached a similar level as in the maternal circulation. Striking was the difference in results for IgG2 with a slow linear rise throughout gestation. The fetal IgG2 level at term remained significantly below the maternal concentration. The IgG subclasses when characterized according to the differences in transport capacity gave the following sequence: IgG1>IgG4>IgG3>IgG2. Fetal IgA showed a slow linear rise with fetal levels at term remaining approximately 1,000 times lower than the concentration in the maternal circulation.

CONCLUSIONS: Comparison of fetal and maternal levels of Immunglobulines indicate that the human placenta during pregnancy develops a specific transport mechanism for IgG. There are differences for the four subclasses with preferential transfer of IgG1 while the slowest transfer is seen for IgG2.
Sperm, Nuclear, Phospholipid, and Red Blood Cell Antibodies and Isotype RF in Infertile Couples and Patients With Autoimmune Rheumatic Diseases
Authors: Fichorova R. , Nakov L. , Baleva M. , Nikolov K. , Gegova I.
Published: September 2011

PROBLEM: To determine if measuring of nonorgan‐specific autoantibodies is useful for better understanding and management of unexplained infertility.

METHODS: Sera were obtained from 70 infertile couples, 57 rheumatic patients, and 76 fertile donors. Sperm antibodies (SA) were detected by the tests of Kibrick and Friberg, anti‐histones, anti‐cardiolipin antibodies, and RF isotypes by ELISA, antinuclear antibodies by indirect immunofluorescence, and anti‐red blood cell antibodies by Capture‐R.

RESULTS: Multiple autoimmune reactivity (both partners positive and/or more than one type of autoantibody involved), higher than naturally occurring in fertile individuals, was found in 55% of the idiopathically infertile couples. IgA‐RF was the dominant autoimmune marker. SA revealed similar rates in patients with rheumatic diseases and in infertiles with or without other autoantibodies.

CONCLUSION: Although no single autoimmunity marker could predict occurrence of SA, the coincidence of enhanced polyclonal autoimmunity in both partners of infertile couples might potentiate their negative effect on reproduction.
Immunotherapy for Recurrent Pregnancy Loss: Analysis of Results From Clinical Trials
Authors: Coulam Carolyn B. , Stephenson Mary , Stern J. Jaros , Clark David A.
Published: September 2011

PROBLEM: Up to 80% of unexplained recurrent spontaneous abortions (RS A) are thought to have an immunologic mechanism. Yet clinical trials using immunotherapy to treat women experiencing RSA have low treatment effects. The present study was undertaken to explain the low treatment effects.

METHODS: Results of clinical trials using allogeneic leukocyte immunization and intravenous (IV) immunoglobulin (Ig) are compared. The mechanisms of pregnancy loss are reviewed in light of data on frequency of karyotype abnormalities in trophoblast of failing pregnancies.

RESULTS: Results of two independent analyses using allogeneic leukocyte immunization as immunotherapy for all women with RSA revealed live birth ratios of 1.16 (P = 0.03) and 1.21 (P = 0.02). When the analysis was limited to primary aborters, the live birth ratio increased to 1.46 (P = 0.006). Live birth ratio after immunotherapy for all RSA using IVIg was 1.88 (P = 0.04). Because of low treatment effects, confounders to treatment success of maternal age and number of previous abortions were studied. Chromosomal abnormalities have been identified in 55% of concepti from RSA. The frequency of chromosomal abnormalities remained constant for up to six pregnancy losses. Women with a history of primary compared to secondary RSA had a higher frequency of karyotypically abnormal concepti (χ2 = 4.54, P < 0.05). Risk factors for RSA also include number of previous losses.

CONCLUSION: Chromosomal abnormalities are a significant confounder when evaluating efficacy of immunotherapy for treatment of RSA. Some women with RSA have a high risk of recurrent chromosomal problems.
Intravenous Immunoglobulin in Women With Five or More Abortions
Authors: Carp H.J.A. , Ahiron R. , Mashiach S. , Schonfeld Y. , Gazit E. , Toder V.
Published: September 2011

PROBLEM: Treatment for recurrent miscarriage has usually been given to all women with three or more abortions of unknown cause. As these patients have a 50–60% subsequent live birth rate, no treatment has been shown to unequivocally improve the live birth rate. Immunoglobulin is the latest treatment to be applied. In order to determine if immunoglobulin improves the live birth rate, we analyzed the results of patients expected to have a poor outcome in the subsequent pregnancy if left untreated, i.e., women with five or more abortions, who have aborted after paternal leucocyte immunization or who continue to abort despite possessing anti‐paternal complement dependent antibody (APCA).

METHODS: A preliminary trial was carried out using immunoglobulin (Sandoglobulin, Sandoz, Switzerland). It was infused at a dose of 400mg/Kg body weight, in the follicular phase of a cycle in which pregnancy was planned. A booster dose was administered as soon as pregnancy was diagnosed.

RESULTS: Twelve patients were treated, ten conceived. Five have had subsequent live births. Two infants were premature but their size was appropriate for gestational age. The other three infants delivered at term.

CONCLUSIONS: This is still too small a group from which to draw definite conclusions about the efficacy of immunoglobulin to prevent abortion. However, five live births in ten patients is an encouraging result, especially when the expected poor obstetric outcome is considered. Hence the efficacy of immunoglobulin should be evaluated further in high risk patients.
Intravenous Immunoglobulin Inhibits Natural Killer Cell Activity In Vivo in Women With Recurrent Spontaneous Abortion
Authors: Ruiz Jaime E. , Kwak Joanne Y.H. , Baum Linda , Gilman‐Sachs Alice , Beaman Kenneth D. , Kim Yoon B. , Beer Alan E.
Published: September 2011

We previously reported elevation of natural killer (NK) cells in women with recurrent spontaneous abortion (RSA) of immune etiology. In this study, we investigated the effect of intravenous immunoglobulin G (IVIg) on peripheral blood NK activity in vivo in women with RSA. Blood was drawn prior to and 7–11 days after IVIg therapy in eight women with RSA. NK activity was measured using K562 as target cells for 51Cr‐release assays. Serum IgG concentrations were also measured. All received 400 mg/kg/day of IVIg for 3 consecutive days. 1) Seven of eight women became pregnant. Five delivered a live born infant. Three out of five women (60%) who delivered a live born infant showed a significant inhibition of NK cytotoxicity post IVIg and the rest did not show any changes; 2) NK cytotoxicity was significantly increased in a woman who miscarried again; 3) A woman who miscarried a chromosomally abnormal fetus showed a significant inhibition of NK cytotoxicity after IVIg; and 4) Serum IgG concentration increased significantly from 9.3 ± 3.0 mg/ml to 23.5 ± 5.1 mg/ml post IVIg therapy. IVIg effectively inhibits peripheral blood NK activity in vivo. These results are consistent with our previous finding showing that IVIg inhibits NK cell activity in vitro. Women with RSA and elevated NK cells may benefit from IVIg treatment.
A Clinical Trial for the Treatment of Antiphospholipid Antibody‐Associated Recurrent Pregnancy Loss With Lower Dose Heparin and Aspirin
Authors: Kutteh William H. , Ermel LeAnn D.
Published: September 2011

This study was conducted to determine if lower dose heparin (LD Heparin) combined with aspirin is as efficacious as higher dose heparin (HD Heparin) for the treatment of the antiphospholipid antibody syndrome in women seeking pregnancy. The method of the study was a prospective, single center trial including 50 patients who were consecutively assigned to treatment. Each patient had at least three consecutive, spontaneous pregnancy losses, positive antiphospholipid antibodies on two occasions, and a complete evaluation. Data were compared using Fisher's exact test. Viable infants were delivered from 20/25 (80%) women treated with higher dose heparin vs. 19/25 (76%) of women treated with lower dose heparin. There were no significant differences between groups with respect to gestational age at the time of delivery (37.2 ± 3.4 versus 37.7 ± 1.6 weeks), maternal complications, or fetal complications. A lower dose of heparin plus aspirin was as effective as higher dose heparin for the treatment of antiphospholipid antibody‐associated recurrent pregnancy loss.