bluuebunny/crossref_metadata_2025_split huggingface dataset:
Direct Production of the Fab Fragment Derived From the Sperm Immobilizing Antibody Using Polymerase Chain Reaction and cDNA Expression Vectors
Authors: SAWAI HIDEAKI , YAMASAKI NORIYUKI , SHIGETA MINORU , KOMORI SHINJI , KARASUYAMA HAJIME , KOYAMA KOJI , ISOJIMA SHINZO 
Published: May 2013

PROBLEM: Sperm immobilizing antibodies cause infertility mainly through complement dependent sperm immobilization. To analyze any effect of sperm immobilizing antibody on fertilization, we had already established cell lines that secrete IgM monoclonal antibody (MAb H6‐3C4) and IgG monoclonal antibody (MAb EnBCMGS). The latter was a class‐switched recombinant IgG antibody that shares the same variable region as MAb H6‐3C4. The biological effects of the IgG antibody were also reported previously to eliminate sperm immobilizing or sperm agglutinating activities. However, the method of chemical digestion of IgG had some disadvantage to prepare the purified Fab fragment stably and in large quantities. This time we report a unique method to obtain the recombinant Fab fragments (Fab EnBCMGS) using polymerase chain reaction (PCR) and cDNA expression vectors.METHOD: Two kinds of PCR primers were designed to make a truncated heavy chain (Fd) gene of MAb EnBCMGS. The amplified Fd gene and light chain gene were ligated into cDNA expression vectors and then transfected into mammalian cells.RESULTS: Expression of the Fd gene and light chain gene were confirmed by Northern blotting. Secretion of the recombinant Fab fragment from mammalian cells was also confirmed by Western blotting. The Fab fragment showed biological activity as is expected by FACS analysis.CONCLUSION: This method enables the stable production of genuine Fab fragments of IgG in mammalian cells without any chemical treatment that may be time consuming and affect the quality of the Fab fragments.
Comparative Studies on the Effects of Specific Immunoneutralization of Endogenous FSH or LH on Testicular Germ Cell Transformations in the Adult Bonnet Monkey (Macaca radiata)
Authors: SURESH R. , MEDHAMURTHY R. , MOUDGAL N.R. 
Published: May 2013

PROBLEM: It is yet to be determined clearly whether the two hormones FSH and T act synergistically in the same cell type—the Sertoli cells—to control overall spermatogenesis or influence independently the transformation of specific germ cell types during spermatogenesis in the adult mammal.METHOD: Adult male bonnet monkeys specifically deprived of either FSH or LH using immunoneutralization techniques were monitored for changes in testicular germ cell transformation by DNA flow cytometry.RESULTS: FSH deprivation caused a significant reduction (>40%; P < 0.05) in [3H] thymidine incorporation into DNA of proliferating 2C (spermatogonial) cells, a marked inhibition (>50%) in the transformation of 2C to primary spermatocytes (4C) and a concomitant, belated reduction (50%) in the formation of round spermatids (1C). In contrast, specific LH/T deprivation led to an immediate arrest in the meiotic transformation of 4C to 1C/HC leading to an effective and significant block (<90%; P < 0.01) in sperm production.CONCLUSION: Thus, LH rather than FSH deprivation has a more pronounced and immediate effect as the former primarily blocks meiosis (4C → 1C/HC) which controls production of spermatids. These data provide evidence for LH/T and FSH regulating spermatogenic process in the adult primate by primarily acting at specific germ cell transformation steps.
Immunohistochemical Localization of C9 Neoantigen and the Terminal Complement Inhibitory Protein CD59 in Human Endometrium
Authors: RATNOFF WILLIAM D. , BROCKMAN WENDY W. , HASTY LISA A. 
Published: May 2013

PROBLEM: Human endometrium expresses complement components, receptors, and regulatory proteins, many of which appear to be expressed in a hormone‐dependent manner. Whether terminal complement components are also present in the endometrium is unknown. CD59, a broadly expressed protein that blocks association of C9 with C8 in the membrane attack complex, is localized in reproductive tissue to human spermatozoa, seminal plasma, amniotic fluid, and placenta. The present study examines human endometrium for the presence of CD59 and terminal complement proteins.METHOD: Endometrial biopsies were obtained from six normal women from various phases of the menstrual cycle and analyzed by immunohistochemistry, using MEM‐43 anti‐human CD59 and anti‐human SC5b‐9 murine monoclonal antibodies and the immunoperoxidase technique.RESULTS: Both CD59 protein and SC5b‐9 (C9 neoantigen) were demonstrated to be present in endometrial glandular epithelium throughout the menstrual cycle. No specific staining was demonstrated in the stromal compartment.CONCLUSION: CD59 protein and terminal complement proteins are expressed in glandular epithelial cells of normal human endometrium, in both proliferative and luteal phases, suggesting that expression is not hormonally dependent. These analyses further support the presence of a functionally active complement system in normal human endometrium.
Up‐Regulated Expression of CD56+, CD56+/ CD16+, and CD19+ Cells in Peripheral Blood Lymphocytes in Pregnant Women With Recurrent Pregnancy Losses
Authors: KWAK JOANNE Y.H. , BEAMAN KENNETH D. , GILMAN‐SACHS ALICE , RUIZ JAIME E. , SCHEWITZ DAVID , BEER ALAN E. 
Published: May 2013

PROBLEM: To analyze immunophenotypic profiles of peripheral blood and humoral autoimmune responses in women with a history of recurrent spontaneous abortions (RSA).METHOD: Peripheral blood lymphocyte subsets by flow cytometry and autoantibodies to phospholipids and nuclear components by ELISA were measured in non pregnant and pregnant women with RSA of unknown etiology. Thirty‐five pregnant and eighty‐one nonpregnant women with RSA were studied. Seventeen nonpregnant and twenty‐two pregnant normal controls were included.RESULTS: Natural killer (NK) cells (CD56+) were significantly elevated in nonpregnant women with RSA as compared with nonpregnant controls. Pregnant women with RSA demonstrated significantly increased NK (CD56+, CD56+/CD16+) and B cells (CD19+) as compared with pregnant controls. Women who miscarried the index pregnancy demonstrated significantly lower CD3+ cells in comparison with normal controls. Women with RSA and antiphospholipid antibodies showed significantly elevated NK cells when compared with women without antiphospholipid antibodies. Women with autoantibodies to nuclear components demonstrated significantly elevated CD19+/CD5+ cells when compared to women without autoantibodies to nuclear components.CONCLUSIONS: Women with RSA demonstrate an abnormal cellular immune response by increasing peripheral natural killer cells and B cells as compared with normal controls.
Anamnestic Development of Lymphocytic Infiltration in Murine Experimental Autoimmune Oophoritis Is Primarily Localized in the Stroma and Theca
Authors: RAJ SHAILAJA , NAIR SHARMILA , MASTORAKOS GEORGE , NELSON LAWRENCE M. 
Published: May 2013

PROBLEM: Neonatal thymectomy performed on day 3 of life (NTX3) induces autoimmune oophoritis and ovarian failure in B6A mice. These mice develop high‐titer autoantibodies specific to oocytes, and ultimately the ovaries become fibrotic and devoid of primordial follicles. These findings implicate the oocyte as a primary target of the autoimmune process. However, in previous work we demonstrated that in developing disease the lymphocytic infiltration was confined to the stroma and theca, and not found involving oocytes. Here, we investigate the possibility that lymphocytic infiltration involving oocytes develops as part of end‐stage disease.METHOD: We transplanted normal syngeneic ovaries to B6A mice with confirmed autoimmune ovarian failure, and, as a control, to normal oophorectomized mice. We then defined the time course and histologic distribution of lymphocytic infiltration in the transplanted ovaries. Lymphocytes were identified by morphology with the aid of an immunohistochemical leukocyte marker (CD45).RESULTS: Autoimmune oophoritis developed by 7 days after transplantation to the NTX3 mice. Compared to control mice, in these mice we found significantly increased stromal and thecal lymphocytic infiltration. In no case did we observe lymphocytic infiltration involving oocytes.CONCLUSIONS: Our findings agree with our previous report and suggest that the ovarian failure in this model is not mediated by a direct lymphocytic attack against intact oocytes. Other immune‐mediated mechanisms are responsible. The paradoxical development of high‐titer oocyte‐specific antibodies despite the stromal and thecal location of the lymphocytic infiltration remains to be explained.
Murine Experimental Autoimmune Oophoritis Develops Independently of Gonadotropin Stimulation and is Primarily Localized in the Stroma and Theca
Authors: NAIR SHARMILA , MASTORAKOS GEORGE , RAJ SHAILAJA , NELSON LAWRENCE M. 
Published: May 2013

PROBLEM: Neonatal thymectomy performed on day 3 of life (NTX3) induces experimental autoimmune oophoritis in certain strains of mice. The disease has its onset around the time of the first estrous, suggesting the process may be gonadotropin dependent. Furthermore, one study reported that gonadotropin stimulation exacerbated the ovarian lymphocytic infiltration in NTX3 mice. Here we examine the possibility that gonadotropin stimulation of the ovary plays a role in the development of post‐thymectomy autoimmune oophoritis.METHOD: Using immunohistochemistry we defined the time course and histologic distribution of the post‐thymectomy ovarian lymphocytic infiltration that develops in B6A mice ([C57BL6 × A/J]F1). We detected ovarian leukocytes using a monoclonal antibody against mouse CD45/T200 and counted those positive staining cells that had the morphologic appearance of lymphocytes. We then treated NTX3 mice to determine if gonadotropin stimulation could exacerbate the disease or cause the disease to appear earlier. We also treated NTX3 mice to determine if gonadotropin suppression could reduce the severity of the disease.RESULTS: Ovarian lymphocytic infiltration was observed as early as 3 weeks after thymectomy, and, during the course of the disease, was primarily located in the stroma and theca. Gonadotropin stimulation did not exacerbate existing disease or induce an earlier onset of severe disease. Furthermore, gonadotropin suppression did not reduce the degree of lymphocytic infiltration or oocyte destruction.CONCLUSIONS: Our findings suggest that murine experimental autoimmune oophoritis develops independently of gonadotropin stimulation of the ovary.
Low‐Dose Aspirin Therapy and Placental Pathology in Women With Poor Prior Pregnancy Outcomes
Authors: CUSICK WILLIAM , SALAFIA CAROLYN M. , ERNST LINDA , RODIS JOHN F. , CAMPBELL WINSTON A. , VINTZILEOS ANTHONY M. 
Published: May 2013

PROBLEM: To determine if low dose aspirin therapy improves placental histology in women with a prior complicated pregnancy demonstrating uterine vascular pathology.METHOD: A retrospective chart review identified patients with a prior complicated pregnancy with placental changes showing uterine vascular pathology (control pregnancy, CP). In the treated pregnancy (TP), 81 mg/day of ASA was started prior to 10 weeks. Placental reports from the CP and TP were reviewed. Pregnancy outcomes and placental histology from the CP were compared to the TP for each patient.RESULTS: Thirteen patients were enrolled. The majority of patients (8/13, 61.5%) exhibited recurrent, histologic evidence of uterine vascular pathology in the TP. The TP was more likely to be uncomplicated (P < 0.05), delivered after 36 weeks (P < 0.05), and result in the delivery of a viable infant (P < 0.05) compared to the CP.CONCLUSIONS: Despite an improvement in outcomes in the aspirin treated pregnancy, histologic evidence of uterine vascular pathology persisted in the majority of women with a prior complicated pregnancy demonstrating similar placental lesions. Abnormal placental histology may be useful in identifying a group of women with poor obstetrical histories who could benefit from low‐dose aspirin therapy.
Amniochorion: A Source of Interleukin‐8
Authors: FORTUNATO STEPHEN J. , MENON RAMKUMAR , SWAN KENNETH F. 
Published: May 2013

PROBLEM: Preterm labor and premature rupture of the membranes are major complications of pregnancy. We have reported the possible role of amniochorionic membrane in the production of inflammatory cytokines and the early onset of labor. This study was conducted to detect the expression of IL‐8 mRNA and peptide production in cultured fetal membranes.METHOD: Amniochorionic membranes were collected from women undergoing elective cesarean section at term. Membranes were cultured in an organ explant system and the expression of IL‐8 was studied over a 10‐day period by RT‐PCR and in situ hybridization. IL‐8 peptide localization was accomplished using immunocytochemistry.RESULTS: Constitutive expression of IL‐8 mRNA in cultured fetal membranes was demonstrated in both amniotic and chorionic leave cells. mRNA and peptide for IL‐8 was homogeneously distributed throughout the amniotic and chorionic cells.CONCLUSION: Human amniochorionic membrane is a source of IL‐8 mRNA and peptide.
γδ T‐Cell Subset Distribution in Human Semen From Fertile Donors
Authors: BERTOTTO A. , SPINOZZI F. , GERLI R. , BASSOTTI G. , FABIETTI G.M. , CASTELLUCCI G. , VAGLIASINDI C. , VACCARO R. 
Published: May 2013

PROBLEM: γδ T‐cell subset distribution has not been fully investigated in normal human semen.METHODS: We therefore carried out experiments by using a direct immunofluorescence staining technique followed by two‐color cytofluorimetric analysis on mononuclear cell (MC) suspensions from ejaculates of ten healthy, fertile volunteers. Autologous peripheral blood MC were simultaneously analyzed and the results used for statistical comparison.RESULTS: The proportion of normal human semen lymphocytes bearing the γδ T‐cell receptor for antigen was greatly increased compared with autologous circulating counterparts. Interestingly, the rise was mainly due to an overexpansion of cells expressing Vδl gene‐encoded determinants on their surface. This contrasts with the normal blood picture, where most γδ T cells express Vδ2 conformational epitopes.CONCLUSIONS: The numerical and phenotypical differences in semen γδ T lymphocytes provide further evidence of a defined migrating lymphocyte subset balance in anatomically and physiologically distinct areas of the body. Their functional role, in terms of both helper and suppressor‐cytotoxic activities in the nonsterile proximal portions of the male genital tract, now needs to be explored in detail.
Immunohistochemical Localization of Endothelin‐1 in Placenta and Fetal Membranes in Term and Preterm Human Pregnancy
Authors: MARINONI EMANUELA , PICCA ARIANNA , SCUCCHI LUIGI , COSMI ERMELANDO V. , DI IORIO ROMOLO 
Published: May 2013

PROBLEM: The aim of the study was to determine the ET‐1 localization on human placenta and fetal membranes and to compare its distribution between term and preterm pregnancies in laboring and non‐laboring tissues.METHODS: Tissues obtained from nine term elective cesarean section, eight spontaneous vaginal term delivery, and 13 preterm delivery from both cesarean section (N = 6) and vaginal delivery (N = 7) were studied by immunohistochemistry.RESULTS: Immunoreactive ET‐1 (IR‐ET‐1) was detected in villous and nonvillous trophoblast in all groups, although laboring tissues showed strong staining in the syncytiotrophoblast of the villi. ET‐1 immunostaining of endothelial cells was observed in all placental villous vessels with a considerable variability within groups. In the fetal membranes, intensive immunopositive staining was observed in the chorionic trophoblast following vaginal deliveries in term and preterm tissues.CONCLUSIONS: This is the first study to report the localization of IR‐ET‐1 in human fetal membranes and placenta, and suggests that amnion and trophoblast represents a source of ET‐1 production or, alternatively, a site for ET‐1 binding.
Normal Uterine Cervix: Characterization of Isolated Lymphocyte Phenotypes and Immunoglobulin Secretion
Authors: CROWLEY‐NOWICK PEGGY A. , BELL MARIA , EDWARDS ROBERT P. , MCCALLISTER DANIEL , GORE HAZEL , KANBOUR‐SHAKIR AMAL , MESTECKY JIRI , PARTRIDGE EDWARD E. 
Published: May 2013

PROBLEM: Isolation of viable cervical lymphocyte populations and characterization of their function in healthy tissue is necessary to understand immunity in the genital tract.METHODS: Normal, cervical tissue was digested using a multi‐enzymatic digestion procedure. Lymphocytes were characterized using FACS analysis and ELISPOT analysis for immunoglobulin secreting cells.RESULTS: Following the digestion procedure, 0.16 × 106 ± 0.8 cells/g of tissue with a viability of 90–98% were isolated from normal cervical tissue. FACS analysis determined that B lymphocytes were the predominant cell type in normal cervical tissue representing a significantly higher percentage than that found in peripheral blood (P=0.015). T lymphocytes and NK cells represented a significantly lower percentage than that found in peripheral blood (P=0.0001 and 0.026, respectively). The largest percentage of immunoglobulin secreting cells isolated were secreting IgG followed by IgA. A limited number of IgM secreting cells were detected. IgA2 secreting cells represented 34.46 ± 4.6% of the total number of IgA plasma cells.CONCLUSION: These studies represent the first analysis of viable mononuclear cells isolated from normal cervical tissue. The results form a baseline from which it will now be possible to compare changes that occur at the cervical squamocolumnar junction in response to infection or neoplasia.
Anticardiolipin Antibodies in Patients With Pregnancy Loss Induce Factor Xa Production in the Presence of (β2‐Glycoprotein I
Authors: OGASAWARA MAYUMI , AOKI KOJI , MATSUURA EIJI , KUNIMATSU MITOSHI , OHKUBO IWAO , GALLI MONICA , SASAKI MAKOTO , YAGAMI YOSHIAKI 
Published: May 2013

PROBLEM: Anticardiolipin antibodies (aCL) are commonly associated with recurrent pregnancy loss, though the mechanism is uncertain. Some investigators have indicated that aCL may be directed at a complex made up of cardiolipin and a blood anticoagulant, β2‐glycoprotein I (β2GPI). We therefore investigated the effects of β2GPI‐dependent aCL IgG enriched fractions, isolated from sera of patients with pregnancy losses, on blood coagulation.METHOD: β2GPI‐dependent aCL were prepared from sera of three women with second trimester pregnancy losses, by cardiolipin affinity column chromography, following by anti‐β2GPI affinity column chromatography. The effects of β2GPI and β2GPI‐dependent aCL on the activation of factor X in vitro were examined.RESULTS: β2GPI inhibited the activation of factor X and β2GPI‐dependent aCL blocked this inhibitory effect in a dose dependent manner.CONCLUSION: These results imply the possibility of β2GPI‐dependent aCL induce hypercoagulation or thrombus by blocking the inhibitory effect of β2GPI on activation of factor X, which may result in pregnancy loss.
Immunoteratology: I. MHC Involvement in the Embryo Response to Teratogens in Mice
Authors: TORCHINSKY A. , FEIN A. , TODER V. 
Published: May 2013

PROBLEM: The present study was carried out to evaluate an involvement of MHC‐associated maternal immunoreactivity in response to environmental teratogens.METHODS: Two chemicals, cyclophosphamide (CP) and 2,3‐quinoxalinedimetanol, 1,4‐dioxide (QD) were used as the reference teratogens (RT). The response to these RT was investigated in syngeneically and allogeneically mated CBA/J and C57B1/6 mice. In part of C57B1/6 female mice, paraaortic lymph nodes were extirpated 14 days before mating to allogeneic or syngeneic males. Twenty or 40 mg/kg of CP or 300 or 600 mg/kg of QD were injected on day 12 and 9 of pregnancy, accordingly (vaginal plug indicates day 1 of pregnancy). On day 19 of pregnancy implantation sites, resorptions, live and dead fetuses were recorded and live fetuses were examined with methods routinely used in applied teratology.RESULTS: Both mice strains showed equal response to teratogens but the RT‐induced effect was significantly weaker in allogeneic than syngeneic mouse combinations. Extirpation of draining lymph nodes dramatically increased the sensitivity to RT in allogeneically mated females but failed to alter that of syngeneically mated ones.CONCLUSION: The results of this study suggest that fetomaternal MHC incompatibility exerts the favourable influence on teratological resistance of the embryo and MHC‐associated immunoreactivity of “mother‐fetus” axis is possibly responsible for this effect.
Detection of Monocyte Chemotactic and Activating Factor (MCAF) and Interleukin (IL)‐6 in Human Seminal Plasma and Effect of Leukospermia on These Cytokine Levels
Authors: SHIMOYA KOUICHIRO , MATSUZAKI NOBORU , IDA NOBUO , OKADA TAKAYOSHI , TANIGUCHI TAKESHI , SAWAI KEISUKE , ITOH SHINICHI , OHASHI KAZUTOMO , SAJI FUMITAKA , TANIZAWA OSAMU 
Published: May 2013

PROBLEM: To demonstrate whether monocyte chemotactic and activating factor (MCAF) and interleukin‐6 (IL‐6) are present in the seminal plasma, and whether these presence is modulated by leukospermia.METHODS: Semen samples from 53 men were obtained by masturbation and examined for the presence of MCAF and IL‐6 by enzyme immunoassay (EIA). Semen samples were obtained from 28 infertile men without leukospermia, 16 infertile men with leukospermia, and nine proven‐fertile men. The correlation between the amount of MCAF in the seminal plasma with some spermiogram parameters and other cytokines such as IL‐6 and IL‐8 was statistically evaluated.RESULTS: Immunoreactive MCAF was detected in the seminal plasmas of all 53 subjects. The MCAF titer in the seminal plasma of patients with leukospermia (11.19 ± 2.75 μg/1) was significantly higher than that in the seminal plasma of the patients without leukospermia (3.24 ± 0.53 μg/1) and the fertile men (2.78 ± 0.35 μg/1) (P < 0.001). The IL‐6 titer in the seminal plasma of the patients with leukospermia (21.05 ± 4.49 ng/1) was also significantly higher than that in the seminal plasma of the patients without leukospermia (8.77 ± 1.92 ng/1) and the fertile men (6.94 ± 1.27 ng/1) (P < 0.01). There was a high degree of correlation among the levels of MCAF, IL‐6 and IL‐8 in the seminal plasma.CONCLUSIONS: These findings demonstrated the presence of MCAF and IL‐6 in the seminal plasma, and that the levels of these cytokines were elevated in the seminal plasma of the infertile patients with leukospermia.
Prevalence of Embryotoxic Factor in Sera From Women With Unexplained Recurrent Abortion
Authors: THOMASON EDWARD J. , ROUSSEV ROUMEN G. , STERN J. JAROSLAV , COULAM CAROLYN B. 
Published: May 2013

PROBLEM: The presence of embryotoxic factors in sera from women with recurrent spontaneous abortion (RSA) has been proposed as a basis for classification of unexplained RSA. To determine the prevalence of circulating embryotoxins among women with idiopathic RSA, sera from 160 women were studied using the mouse blastocyst assay.METHODS: Two‐cell embryos were collected from superovulated mated CB6F1/J mice and cultured in media supplemented with fetal bovine serum (FBS) or 10% serum at 37°C with 5% CO2 and high humidity. Each assay was run in triplicate using three mice with at least five embryos from each mouse. Results were determined by calculating the average percentage atresia for each mouse. FBS, known to support embryo proliferation, was used to control in each assay.RESULTS: The prevalence of embryotoxic factors among women experiencing RSA was 24.4% (39/160). There is no correlation found between the presence of embryotoxicity and phospholipid antibodies, lupus anticoagulant, and thyroglobulin/microsomal antibodies.CONCLUSION: The embryotoxicity assay can serve as a basis for a new approach for classification of unexplained recurrent spontaneous abortion.