bluuebunny/crossref_metadata_2025 huggingface dataset:
Does Endothelin‐1 Affect Human Spermatozoa Function?
Authors: Kamada Shusaku , Oehninger Sergio , Mahony Mary C. , Blackmore Peter F. , Lanzendorf Susan E. , Hodgen Gary D.
Published: May 2013

PROBLEM: It has been reported that massive amounts of immunoreactive endothelins (ETS) exist in human seminal plasma. However, the physiological role of ETS in seminal plasma remains to be determined. We speculated that ETS might affect sperm function.

METHOD: The present study was designed to investigate the effect of endothelium‐1 (ET‐1) on: (a) sperm motion parameters, (b) hyperactivated motility, (c) sperm‐zona pellucida‐binding capacity, (d) sperm‐oocyte penetration capacity, (e) acrosome reaction and its prerequisite, an increase in intracellular Ca2+ concentration ([Ca2+]i), and to examine (f) the presence of binding sites for ET‐1 in human sperm. Forty‐six semen ejaculates from 14 fertile men were assessed under capacitating conditions after separation of the motile sperm fraction by wash and swim‐up.

RESULTS: ET‐1 (1 (μm) exhibited significant stimulatory effects on sperm velocity at 30 min (P = 0.01), amplitude of lateral head displacement (ALH)max (P = 0.05), and ALHmean at 60 min (P = 0.04) in some samples (n = 10). However, these effects were not observed in experiments using a larger number of samples (n = 39). ET‐1 had no effect on hyperactivated motility of sperm at 30 min to 24 h. Neither ET‐3 nor IRL 1620, a selective ETB receptor agonist, affected sperm motion parameters or hyperactivated motility. ET‐1 did not affect sperm‐zona‐binding capacity, sperm‐oocyte penetration capacity, acrosome reaction, or [Ca2+]i of sperm. Specific binding sites for ET‐1 were not detected on the cell surface of human sperm.

CONCLUSIONS: Although ET‐1 is present in massive amounts in human seminal plasma and may have the capacity to alter the quality of motile sperm in some samples, a physiological role of ET‐1 in the modulation of the function of mature, ejaculated sperm still remains unknown.
Direct Ovarian Effect of Growth Hormone in the Rabbit
Authors: Ando Motomu , Yoshimura Yasunori , Iwashita Mitsutoshi , Oda Takahisa , Karube Masataka , Ubukata Yoshinobu , Jinno Masao , Nakamura Yukio
Published: May 2013

PROBLEM: This study was undertaken to assess whether growth hormone (GH) can stimulate follicle growth and ovarian steroidogenesis via putative GH receptors.

METHOD: In vitro perfused rabbit ovary.

RESULTS: Ovulation occurred in neither the control ovaries nor experimental ovaries treated with 100 ng/ml of GH, whereas all ovaries exposed to 50 IU of human chorionic gonadotropin (hCG) ovulated. The addition of GH to the perfusate significantly stimulated the follicle growth in the absence of gonadotropin. The percent change in follicle diameter in GH‐treated ovaries did not differ significantly from that in hCG‐treated ovaries. Exposure to GH significantly stimulated the meiotic maturation in the follicular oocytes, as compared with the contralateral control ovaries. Although the concentration of progesterone in the perfusate did not differ significantly between GH‐treated and control ovaries, GH stimulated estradiol production by the perfused rabbit ovaries. Rabbit ovary membranes exhibited high affinity binding sites of hGH (Kd = 6.1 × 10−9M).

CONCLUSION: GH acts on the rabbit ovary to stimulate the follicle growth, oocyte maturation, and ovarian estradiol production by interacting with the specific receptors located in ovarian plasma membranes.
The Frequency of Antisperm Antibodies in the Cervical Mucus of Women With Poor Postcoital Tests and Their Effect on Pregnancy Rates
Authors: Check J.H. , Bollendorf A. , Katsoff D. , Kozak J.
Published: May 2013

PROBLEM: To determine the impact of the presence of antisperm antibodies (ASAs) in the cervical mucus of female partners in couples with unexplained poor postcoital tests (PCT). Furthermore, the efficacy of intrauterine insemination (IUI) in these same patients was determined by pregnancy rates (PRs).

METHOD: Pregnancy rates following IUI in patients with infertility and poor postcoital tests, whether the cervical mucus was positive or negative for ASAs, were evaluated.

RESULTS: The 6‐month PRs were similar in the ASA negative (40.5%) versus the positive (42.4%) group.

CONCLUSIONS: It appears that the antifertility effect of ASA may be mainly the immobilization of sperm in the cervical mucus, and thus, performing IUI may effectively correct the problem.
Immunization of Mice Against a Synthetic N‐Terminal Extracellular Domain Gonadotropin‐Releasing Hormone Receptor Peptide: Evidence for a Direct Uterine Effect
Authors: Asirvatham Angela L. , Johnson Gregory A. , Belden E. Lee , Kirk Edward A. Van , Moss Gary E. , Murdoch William J.
Published: May 2013

PROBLEM: Immature male and female mice were immunized with a synthetic peptide corresponding to amino acids 5–17 (ASLEQDPNHCSAI) of the mouse hypophyseal gonadotropin‐releasing hormone (GnRH) receptor.

METHOD: Effect of immunization (postpuberal) was restricted to the uterus. Pituitary‐go‐nadal functions were not altered.

RESULTS: The endometrial lining of immunized females was thin and lacked glandular development. These observations were corroborated in actively immunized and passively immunized adult females.

CONCLUSIONS: Apparently endometrial cells express a unique surface antigen, though reactive with antipeptide antibodies, that differs from the prototype pituitary GnRH receptor. Antibodies that selectively inhibit endometrial maturation might be used to treat proliferative diseases of the uterus.
Interleukin‐6 Does Not Stimulate Rat Myometrial Contractions in an In Vitro Model
Authors: Dajani Nafisa , Idriss Edo , Collins Patricia L.
Published: May 2013

PROBLEM: Interleukin‐6 (IL‐6) is increased in culture‐positive amniotic fluid in women with preterm labor. IL‐6 stimulates the production of prostaglandins leading to increased uterine activity.

METHODS: We tested the hypothesis that IL‐6 increases myometrial activity through release of uterotonic mediators. We studied the effect of IL‐6 on uterine contractions in the absence and presence of fetal membranes to determine if the effect was on myometrium alone or was mediated through fetal membranes/decidua. IL‐6 in concentrations of 100, 10, 0.1 or 0 ng/ml was added to the maternal side of the dual chamber‐fetal membrane‐uterine muscle in vitro model.

RESULTS: We found that 10 ng/ml of IL‐6 alone, without fetal membranes, caused a significant decrease in uterine contractions over time (P≤0.01). This decrease was not observed with the addition of term, nonlabored fetal membranes.

CONCLUSIONS: IL‐6 in the presence or absence of membranes, over a four log fold dose range, did not stimulate uterine contractions.
Alternative Treatment to Lymphocyte Immunization for Treatment of Recurrent Spontaneous Abortion: Immunotherapy With Intravenous Immunoglobulin for Prevention of Recurrent Pregnancy Loss: European Experience
Authors: Mueller‐Eckhardt Gertrud
Published: May 2013

PROBLEM: Due to its strong “immunomodulating” effect in several well established disorders, high‐dose intravenous immunoglobulins (IVIG) has been proposed as an alternative for immunotherapy with allogeneic leucocytes in patients with unexplained recurrent spontaneous abortion. This paper is intended to provide an overview on the European experience in this field.

METHOD: Five European pilot studies with a total of 172 patients as well as one controlled double‐blind multicenter study including 64 patients were considered. In the latter, 5% human albumin was used as placebo.

RESULTS: Success rates of the pilot studies varied from 68 to 87%. In the German controlled study, a significant specific effect of IVIG could not be verified. However, success rates for both IVIG and albumin were in the same range as for allogeneic leucocytes.

CONCLUSION: At present, it is not sufficiently proven that IVIG is an appropriate tool for immunotherapy of recurrent spontaneous abortions. It is suggested that success rates of both IVIG and albumin are due to a placebo effect. However, we cannot exclude that albumin itself provides immunomodulating capacity.
Inhibition of Induced Lymphocyte Proliferation by Lipid and Protein Components of the Syncytiotrophoblast Plasma Membrane
Authors: GIRARD ANNE‐CHRISTINE LALMANACH , GUILLAUMIN JEAN‐MAURICE , THIBAULT GILLES , RUPIN ALAIN , DE RUSSÉ JEANNE , BARDOS PIERRE
Published: May 2013

PROBLEM: The aim of this work was to define the respective responsibilities of the lipid and protein components of syncytiotrophoblast plasma membranes on the inhibition of lymphocyte proliferation induced in vitro.

METHOD: A fractionation method using octyl‐β‐D‐glucopyranoside enabled lipoprotein, lipid, and protein fractions to be isolated from the membrane. RESULTS: The lipid fraction was shown nonspecifically to inhibit lymphocyte proliferation, to a lower extent compared with the native membrane. Alternatively, the protein fraction used as a proteoliposome contained the totality of the cytostatic effect of the native fraction.

CONCLUSION: These results are discussed generally in the context of the immunoregulatory role of membrane lipids and proteins and in relation to the local properties of syncytiotrophoblast plasma membrane components in fetal graft tolerance.
Alloimmunization Against Well Defined Polymorphic Major Histocompatibility or Class I MHC Transfected L Cells Antigens Can Prevent Poly IC Induced Fetal Death in Mice
Authors: MENU ELIZABETH , CHAOUAT GÉRARD , KINSKY RADOSLAV , DELAGE GENEVIÉVE , KAPOVIC MILJENCO , THANG M.N. , JAULIN CHRISTIAN , KOURILSKY PHILIPPE , WEGMANN THOMAS G.
Published: May 2013

METHOD: It is possible to induce increased fetal resorption in a number of inbred murine matings by injecting Poly (I) Poly (C12U) 3.5 days postconception, a maneuver associated with natural killer‐mediated damage to the feto placental unit such as occurs in spontaneous fetal resorptions.

RESULTS: We show here that alloimmunization can block this effect. In addition, maternal immune responses induced by alloimmunization against isolated mutant class I or class II, as well as by immunization with class I MHC alloantigens (Kd) transfected L cells are sufficient to restore normal fetal viability. It is not necessary that the maternal immune response be specifically directed against paternal alloantigens fr the fetal protecton to ensue, since the effect occurs in inbred matings when the mother is immunized against unrelated class I or class II alloantigens. As in previous studies conducted in the murine species, not all MHC alloimmunizations are protective. In addition, as control, immunization with a monomorphic class I MHC molecular (37), transfected L cells, sheep red blood cells or hen egg lysozyme is without effect.

CONCLUSION: These results indicate that defined MHC antigens can mediate fetal protection from induced fetal resorption, and suggest that one driving force in promoting MHC antigen polymorphism in mammals is their capacity to confer protection from NK mediated fetal demise.
Embryo‐Associated Immunosuppressor Factor is Produced at the Maternal‐Fetal Interface in Human Pregnancy
Authors: BOSE RATNA , LACSON ATILANO G.
Published: May 2013

PROBLEM: To study whether embryo associated immunosuppressor factor (EASF) is synthesized at the maternal‐fetal interface.

METHOD: Anti‐EASF monoclonal antibody H5D12 was used to identify EASF. Paraffin‐embedded sections were prepared from placental and fetal tissues and immunohistochemistry was done by the avidin‐biotin‐peroxidase technique. EASF was affinity purified using H5D12‐Sepharose 4B from culture media of placental villi and analyzed for immunosuppressive activity (by Concanavalin A‐induced lymphocyte proliferation assay) and molecular weight identity (by metabolic labeling studies with 35S‐methionine followed by immunoprecipitation and SDS‐PAGE).

RESULTS: Immunohistochemical studies demonstrates intense immunostaining of villous syncytiotrophoblast and cytotrophoblast cells of first trimester placental tissues. Hoffbauer cells and decidual cells stained positive. The same cells in second and third trimester placental tissues stained weakly. However, the endothelium and smooth muscle cells of fetal blood vessels, fetal ovarian stroma and primordial follicles, kidney epithelium, cerebral neurons, and glial cells all stained negative. The affinity‐purified EASF from the conditioned media of placental villi (less than 12 wk gestational age) was identified as a 37‐kDa molecule with immunosuppressive activity. Metabolic labeling studies revealed that placental villi from early gestational age secretes a major factor of 37‐kDa and minor factors of 41‐kDa and 47‐kDa molecular weight.

CONCLUSIONS: Monoclonal antibody H5D12 identifies a factor that is produced by the pre‐implantation embryo and also synthesized by decidua and trophoblast cells.
Sperm Immobilizing Antibodies Interfere With Sperm Migration From the Uterine Cavity through the Fallopian Tubes
Authors: SHIBAHARA HIROAKI , SHIGETA MINORU , TOJI HIDEAKI , KOYAMA KOJI
Published: May 2013

PROBLEM: It is well known that sperm migration in cervical mucus is impaired by sperm immobilizing antibodies secreted in the mucus. However, it is not clear yet whether sperm migration from the uterine cavity through the fallopian tubes to the peritoneal cavity is impaired by sperm immobilizing antibodies. To test the possible impairment of sperm migration in the tubes, laparoscopic examinations were carried out and the presence of motile sperm in the peritoneal fluid after intra‐uterine insemination was investigated.

METHOD: Peritoneal sperm recovery tests were performed in 28 infertile women with sperm immobilizing antibodies in their sera, and the results were compared with those in 322 infertile women without the antibodies. Both the sperm immobilizing antibody titers (SI50) and complement activities (C'H50) in peritoneal fluid were compared with those in patients' sera. In some experiments, the supernatant of the peritoneal fluid was used as a source of complement for the sperm immobilization tests instead of guinea pig serum.

RESULTS: Among couples with normal semen characteristics by the criteria of WHO, sperm recovery in the peritoneal fluid was observed in only 3 (11.1%) of 27 patients with sperm immobilizing antibodies, compared with 72 (34.0%) of 212 patients without the antibodies (P<0.025). The antibody titers of the patients with the sperm recovery were very low by the quantitative sperm immobilization test. In most patients, a similar amount of sperm immobilizing antibodies was present in the peritoneal fluid and the sera. Though the complement activities in the peritoneal fluid were less than those in sera, the former were still found to be sufficient to immobilize sperm in vivo.

CONCLUSIONS: These results suggest that the complement‐dependent sperm immobilizing antibodies could interfere with sperm migration in the female genital tract at the level of the fallopian tubes.
Standardization of Flow Cytometric Crossmatch (FCXM) for Investigation of Unexplained Habitual Abortion
Authors: Stephenson Mary D. , Wu Vivian , Mackinnon Murray , Sadoway Janet , Keown Paul A.
Published: May 2013

PROBLEM: To define a positive flow cytometric crossmatch (FCXM), in terms of channel shift, for maternal IgG and IgM (n=28) against paternal T and B lymphocytes.

METHOD: A reference range study. Mononuclear cells were obtained from 28 healthy volunteers using density gradient separation of heparinized blood, followed by pre‐incubation with goat immunoglobulin. A total of twelve tubes were prepared for each volunteer. Primary incubation was with negative control serum, positive control sera (either IgG or IgM) and individual AB sera. Secondary incubation was with four combinations of fluorochromes; CD3 PE/IgG‐Fc F(ab′)2FITC, CD3 PE/IgM F(ab′)2FITC, CD20 PE/IgG‐Fc F(ab′)2FITC and CD20 PE/IgM F(ab′)2FITC. The cells were then analyzed with an EPICS Profile flow cytometer, using 256‐channels and a four decade log scale.

RESULTS: The linear mean channel fluorescence of the negative control serum was subtracted from the individual AB sera (channel shift) for each of the four combinations of fluorochromes. By determining the 95% one‐sided upper reference limits of the negative control serum for each of the four trimmed data sets, we clinically defined a positive FCXM for bound IgG or IgM to T lymphocytes as a shift of 10 or more channels, and for bound IgG or IgM to B lymphocytes as a shift of 25 or more channels, above the linear mean channel shift of the negative control serum.

CONCLUSION: Positive FCXMs were defined for maternal IgG and IgM against T and B lymphocytes, in terms of channel shift above the linear mean channel fluorescence of the negative control serum. By standardizing the dual‐color FCXM methodology, the clinical significance of alloantibodies in the maintenance of pregnancy could be addressed in a collaborative manner.
A Comparison of Flow Cytometry and Microcytotoxicity for the Evaluation of Alloimmune Therapy in Patients With Recurrent Spontaneous Abortions
Authors: Matzner W. , Chong P. , Xu G. , Ching W.
Published: May 2013

PROBLEM: There is substantial data that support the efficacy of paternal leukocyte immunization (PLI) for the treatment of alloimmune mediated miscarriage; however, there is confusion regarding the laboratory test that should be performed to determine levels of maternal anti‐paternal leukocyte antibodies (MAPLA).

METHOD: Popular methodologies employed include: 1) microcytotoxicity (MCX), 2) mixed lymphocyte culture (MLC), and 3) cell flow cytometry crossmatch (FCXM). Cell flow cytometry crossmatch correlates well with the more difficult MLC assay although the former proves the more sensitive study. This work compares the MCX assays with FCXM. The study group consisted of ten women who had a history of three or more spontaneous abortions (SABs). All ten had very low levels (<10%) of MAPLA as measured by FCXM. Following PLI all subjects demonstrated elevated levels (>50%) of MAPLA by FCXM. At 12 weeks gestation, sera were simultaneously measured for MAPLA by MCX and FCXM.

RESULTS: Although all ten patients had very high levels of MAPLA by FCXM during pregnancy, five of ten had antibodies to HLA Class I and two of ten had antibodies to HLA Class II paternal antigens by MCX. Furthermore, all patients who were positive by MCX to paternal Class I antigens were also positive to Class I antigens not seen in either parent. Both patients who were positive by MCX to paternal Class II antigens were also positive to maternal Class II antigens. Notable is that all ten women eventually delivered healthy infants.

CONCLUSION: Based on this preliminary study, the MCX assay is neither sensitive or reliable enough to determine the need and/or to monitor the effectiveness of PLI. Flow cytometry should be the modality of choice when determining the need for alloimmunotherapy and to monitor the effectiveness of treatment.
Alloantibody Responses to Antigens Recognized by Rabbit Antitrophoblast Antisera in Trophoblast and Mononuclear Cell (MNC) Membranes by Primary Aborting Women Before and After Paternal Leukocyte Immunization
Authors: Goldberg Janet S. , Haynes Mark K. , Cowchock F. Susan , Mcintyre John A. , Smith J. Bruce
Published: May 2013

PROBLEM: Because shared allogeneic antigens are expressed on peripheral blood lymphocytes, as well as trophoblasts, it has been proposed that lymphocyte transfusions may appropriately sensitize recurrent spontaneous aborters (RSA) to trophoblast and lead to pregnancy conservation. The degree to which these responses are affected by this treatment, however, has not been defined.

METHOD: SDS‐polyacrylamide gel electrophoresis (SDS‐PAGE) and Western blot analyses were used to study the alloantibody responses in RSA patients both before and after immunization with paternal leukocytes (MNC) against membrane proteins isolated from peripheral blood mononuclear cells (MNC), first trimester and full term chorionic villi (CV).

RESULTS: A distinct set of antigens with apparent molecular weights of 32, 36, 41, 63, 65, and 85 kDa was identified in both MNC and trophoblast membranes by rabbit anti‐trophoblast antisera. In addition, a 55 kDa protein was recognized by MNC membranes. Thirty‐eight percent of primary RSA sera recognized this 55 kDa protein. After paternal MNC immunization, all primary RSA samples displayed increased reactivity or produced antibodies to this 55 kDa protein when compared with preimmunization sera. The protein was identified as MCP using a rabbit polyclonal anti‐MCP antisera. In contrast, reactivity to the other antigens present in the membrane preparations decreased after paternal MNC immunization.

CONCLUSION: Changes in immune reactivity in RSA patients after paternal MNC immunization suggest that immunization alters serum immune reactivity to MNC and trophoblast shared antigens.
Lymphocyte Subsets and Autoantibodies in Pregnancies Complicated by Placental Disorders
Authors: Matthiesen Leif , Berg Göran , Ernerudh Jan , Skogh Thomas
Published: May 2013

PROBLEM: The survival of the fetoplacental unit might partly depend on down‐regulation of rejection reactions. Pathological maternal cellular immune response mechanisms could therefore be of pathogenic importance in pregnancies complicated by placental disorders.

METHOD: A flow‐cytometric analysis of T‐cell subsets and B‐cell, as well as serological tests for anticardiolipin antibodies (aCL), antinuclear antibodies (ANA), and rheumatoid factor (RF) were done on 90 women with complicated pregnancies. The results were compared with that of nonpregnant women (n=5), and normal pregnant women (n=5) in the third trimester.

RESULTS: Two women, suffering from severe preeclampsia and eclampsia respectively, had aCL on 12 and 13 units, respectively. ANA occurred in 11 patients with moderate and severe preeclampsia and intrauterine growth retardation. All women had negative RF tests. Within the CD4 T‐helper subpopulations, the proportion of suppressor/inducer T‐cell population (CD4+CD45RA+) significantly increased, while the memory and helper/inducer T‐cells (CD4+CD45RO+ and CD4+CD29+) significantly decreased during normal pregnancy compared to nonpregnant controls. This deviation of CD4 subpopulations was not found, or was less pronounced, in complicated pregnancies. The proportion of cytotoxic T cells (CD8+S6F1+) was significantly reduced during normal pregnancy. This reduction was less pronounced in complicated pregnancies.

CONCLUSION: Systemic immunological deviations toward suppression or decreased activity of the immunological response as seen in normal pregnancies, was not observed in preeclampsia, intrauterine growth retardation, intrauterine fetal death, and abruptio placentae. This lack of suppression or increased T‐cell activity may have a primary pathogenic role in some women with a complicated pregnancy, or it may be secondary to the placental disorders.
Systemic CD56+ Cells Can Predict Pregnancy Outcome
Authors: Coulam C.B. , Goodman C. , Roussev R.G. , Thomason E.J. , Beaman K.D.
Published: May 2013

PROBLEM: To evaluate differences in circulating CD56+ cells between successful and unsuccessful pregnancies, 114 pregnant women were studied prospectively.

METHOD: Seventy women had a history of infertility (INF) and 44 had two or more previous spontaneous abortions (RSA). Among the infertile women, 12 were donor egg recipients (DER) and 15 underwent intracytoplasmic sperm injection (ICSI) for treatment of male factor infertility. Nineteen women were carrying multiple gestations (MG) and 55 had singleton gestations (SG). Thirteen additional women were receiving intravenous immunoglobulin (IVIg).

RESULTS: The percentage of CD56+ cells was determined in 310 blood samples from 114 pregnant women by flow cytometry. The prevalence of women with persistent elevation of percent of 56+ cells (>12%) was 58% among DER, 73% among ICSI, 37% among MG, 22% among SG, 18% among RSA, and 39% among INF. Thirteen women with SG received IVIG, 10 had CD56+ cells greater than 12% and all 13 experienced live births. Women with percentage CD56+ cells persistently greater than 12% who were not DER, not ICSI, not receiving IVIg, and not carrying MG had a live birth rate of 11%. Women with greater than 12% CD56+ cells had normal karyotype in 78% of concepti studied in contrast to women less than 12% CD56+ cells who had 68% abnormal karyotypes (P=0.04).

CONCLUSION: Elevated CD56+ cells in pregnant women who are not DER, not ICSI, not receiving IVIg, and not carrying MG predicts loss of a karyotypically normal conceptus with a specificity of 87% and positive predictive value of 78%. While the specificity value of this test is high in both infertile and RSA populations, the sensitivity is 86% in RSA and only 54% in INF suggesting this test does not identify all losses among INF. It may identify a subset of pregnancies at risk for loss of a karyotypically normal embryo that may respond to treatment with IVIg.
Production of Mouse Monoclonal Antibodies Directed Against the Oolemma of Human and Hamster Oocytes by Intra‐splenic Injection of Oocytes
Authors: Fusi F.M. , Gasparri A.M. , Pelagi M. , Tamburini C. , Santis L. De , Grieco S.E. , Siccardi A.G. , Ferrari A.
Published: May 2013

PROBLEM: To develop an additional approach for the study of oolemmal surface moieties involved in gamete interactions, we decided to obtain monoclonal antibodies by intrasplenic injection of human and hamster oocytes in Balb/c mice.

METHOD: Two Balb/c males were injected three times intrasplenically at 15‐day intervals with approximately 40 zona‐free hamster and 3–5 zona‐free human oocytes. After the third injection, spleen cells were fused and hybridomas developed. We used a novel screening system based upon the use of sections of frozen human and hamster eggs, tested by means of indirect immunofluorescence. The antibodies that we produced were evaluated for their ability to interfere with the zona‐free hamster eggs penetration by human spermatozoa. The B2B5 antibody was also developed as ascitic fluid and further characterized.

RESULTS: Seven antibodies reactive with hamster oocytes were produced. Six of them also reacted with human oolemmas. The binding was confined to the oolemma, and no staining of the zona nor the cytoplasm was present. One of these antibodies reduced the penetration of zona‐free hamster eggs by human spermatozoa. This antibody, B2B5, an IgM kappa, was confirmed to interact with the oolemma by means of indirect immunofluorescence of fresh eggs and Covasphere binding. B2B5 did not react with other human or hamster tissues except capacitated human spermatozoa. The reactivity with the oolemma of hamster oocytes was not lost after egg penetration by human sperm.

CONCLUSIONS: Intrasplenic immunization using zona‐free human and hamster oocytes allows the production of anti‐oolemma antibodies. A system of screening based upon the use of sections of frozen eggs also allows an easy and quick scoring of many supernatants. B2B5 monoclonal anti‐oolemma antibody deserves further studies in that is able to interfere with fertilization and its antigen appears to be confined to the gametes surface.
In Vivo CD3+CD25+ Lymphocyte Subpopulation Is Down‐regulated Without Increased Serum‐Soluble Interleukin‐2 Receptor (sIL‐2R) by Gonadotropin Releasing Hormone Agonist (GnRH‐a)
Authors: Ho Hong‐Nerng , Wu Ming‐Yih , Chen Hsin‐Fu , Chao Kuang‐Han , Yang Yu‐Shih , Huang Su‐Cheng , Lee Tzu‐Yao , Gill Thomas J.
Published: May 2013

PROBLEM: To test further whether the suppression of the CD3+CD25+ lymphocyte sub‐population by gonadotropin‐releasing hormone agonist (GnRH‐a) is related to the change in levels of cytokines and soluble interleukin‐2 receptor (sIL‐2R).

METHOD: Twenty‐seven infertile patients were enrolled under the long protocol of GnRH‐a agonist (buserelin acetate) and superovulation with gonadotropin from our IVF‐ET program. Peripheral B cells, NK cells, CD4+ and CD8+ T cells and the expression of CD69, CD25, HLA‐DR, and CD71 antigens on the T cells were serially examined by dual‐color flow cytometry. Serum levels of cytokines and sIL‐2R were measured.

RESULTS: The B cells, NK cells, T cells, CD4+, CD8+ T cells, CD3+DR+, and CD3+CD71+ lymphocyte subpopulations were not changed after the use of GnRH‐a. The CD25+ T cell subpopulation was significantly down‐regulated, but the CD69+ T cell subpopulation was increased when the GnRH‐a was used for approximately 2 wk. The serum levels of interleukin‐lp (IL‐1β), interleukin‐2 (IL‐2), interleukin‐4 (IL‐4), interferon‐γ (IFN‐γ), and sIL‐2R were not changed.

CONCLUSION: The GnRH‐a had a transiently suppressive effect on CD25+ T cells, but a stimulatory effect on CD69+ T cells. However, the serum level of cytokines or sIL‐2R did not change. These immunological modulations seems to be the result of interaction between GnRH‐a and estrogen.
Variation in Antisperm Antibody Results Using Different Assays
Authors: Peters Albert J. , Ivanovic Milica , Jeyendran Rajasingam S.
Published: May 2013

PROBLEM: The detection of various types of antisperm antibodies (ASA) in the serum varies among different assays. This variation may influence the diagnosis and management of infertile couples who are tested for such immunologic factors. This prospective study was conducted to determine the variation in the results of ASA as measured by the sperm immobilization (SI), sperm agglutination (SA), and the indirect immunobead (IB) assays.

METHOD: The sera of 79 patients that tested positive for ASA by at least one of the assays listed above were concurrently tested with all three assays.

RESULTS: Using an individual ASA assay, 66 (84%), 26 (33%) or 36 (46%) of sera tested positive by the SA, SI, or IB assays, respectively. However, using a combination of assays, 67 (85%), 78 (99%) or 40 (51%) of sera tested positive using either the SA+SI, SA+IB or SI+IB assays, respectively.

CONCLUSION: These findings suggest that the utilization of different assays to detect ASA may detect sera that are positive for ASA with more reliability than single assay testing.
Testosterone replacement therapy can increase circulating endothelial progenitor cell number in men with late onset hypogonadism
Authors: Liao C.‐H. , Wu Y.‐N. , Lin F.‐Y. , Tsai W.‐K. , Liu S.‐P. , Chiang H.‐S.
Published: May 2013

Circulating endothelial progenitor cells (EPCs) are bone marrow‐derived cells required for endothelial repair. A low EPC number can be considered as an independent predictor of endothelial dysfunction and future cardiovascular events. Recent evidence shows that patients with hypogonadal symptoms without other confounding risk factors have a low number of circulating progenitor cells (PCs) and EPCs, thus highlighting the role of testosterone in the proliferation and differentiation of EPCs. Here, we investigate if testosterone replacement therapy (TRT) can increase circulating EPC number in men with late onset hypogonadism. Forty‐six men (age range, 40–73 years; mean age, 58.3 years) with hypogonadal symptoms were recruited, and 29 men with serum total testosterone (TT) levels less than 350 ng/dL received TRT using transdermal testosterone gel (Androgel; 1% testosterone at 5 g/day) for 12 months. Circulating EPC numbers (per 100 000 monocytes) were calculated using flow cytometry. There was no significant association between serum TT levels and the number of circulating EPCs before TRT. Compared with the number of mean circulating EPCs at baseline (9.5 ± 6.2), the number was significantly higher after 3 months (16.6 ± 11.1, p = 0.027), 6 months (20.3 ± 15.3, p = 0.006) and 12 months (27.2 ± 15.5, p = 0.017) of TRT. Thus, we conclude that serum TT levels before TRT are not significantly associated with the number of circulating EPCs in men with late onset hypogonadism. However, TRT can increase the number of circulating EPCs, which implies the benefit of TRT on endothelial function in hypogonadal men.
Adapted Physical Activity Programme and Self‐Perception in Obese Adolescents with Intellectual Disability: Between Morphological Awareness and Positive Illusory Bias
Authors: Salaun Laureline , Reynes Eric , Berthouze‐Aranda Sophie E.
Published: May 2013

Background

In adolescent with intellectual disability, the management of obesity is a crucial issue, yet also quite complex because of their particular perception of themselves. This study investigated the relationship between self‐perception variables and morphological variables and their changes after a 9‐month Adapted Physical Activity (APA) programme.

Materials and Methods

Twenty‐three adolescents with intellectual disability responded to an adapted questionnaire, including thePSI‐VSF‐IDand a nine‐drawing body silhouette scale. Anthropometric and body composition indicators were measured before and after theAPAprogramme.

Results

The main predictor of the adolescents' self‐perceptions was the inclination towards positive illusory bias before the intervention; obesity awareness ranked second. Morphological measurements did not contribute in the same way to self‐perceptions in the initial and final data.

Conclusions

This study confirms the interest of weight management programmes for adolescents with intellectual disability and points to the need to take positive illusory bias more fully into account in the study of self‐perception.
Three ileus cases associated with the use of dipeptidyl peptidase‐4 inhibitors in diabetic patients
Authors: Kanasaki Keizo , Konishi Kazunori , Hayashi Ranji , Shiroeda Hisakazu , Nomura Tomoe , Nakagawa Atsushi , Nagai Takako , Takeda‐Watanabe Ai , Ito Hiroki , Tsuda Shin‐ichi , Kitada Munehiro , Fujii Mizue , Kanasaki Megumi , Nishizawa Makoto , Nakano Yasuharu , Tomita Yasuto , Ueda Nobuhiko , Kosaka Takeo , Koya Daisuke
Published: May 2013

Dipeptidyl peptidase (DPP)‐4 inhibitors are a new class of antidiabetic drugs that increase incretin hormone levels to enhance blood sugar level‐dependent insulinotropic effects, suppress glucagon action, and reduce bowel motility. These incretin effects are ideal for blood sugar control. However, the safety profile of DPP‐4 inhibitors is not yet established. Herein, we present three cases of ileus, considered to be closely related to the use of DPP‐4 inhibitors, in diabetic patients. Each of the three patients exhibited some risk of a deficiency in bowel movement; the onset of ileus was within 40 days after strengthened inhibition of DPP‐4. The use of a DPP‐4 inhibitor could be safe, although the cases presented herein enable us to inform the scientific community to some of the potential adverse effects of the use of DPP‐4 inhibitors in select populations.
Treatment of severe autoimmune blistering skin diseases with combination of protein A immunoadsorption and rituximab: a protocol without initial high dose or pulse steroid medication
Authors: Kolesnik M. , Becker E. , Reinhold D. , Ambach A. , Heim M.U. , Gollnick H. , Bonnekoh B.
Published: May 2013

Background

Skin blistering diseases due to autoantibodies are typically treated with high dose systemic corticosteroids and other conventional immunosuppressants. However, in severe cases, this treatment may not be sufficient to achieve disease control or contraindicated because of comorbidity.

Methods

We describe 15 patients (pts.) with such diseases: 6 pts. with pemphigus vulgaris, 3 pts. with bullous pemphigoid, 3 pts. with mucous membrane pemphigoid (MMP), one being anti‐laminin‐332‐MMP (AL332‐MMP), 2 pts. with pemphigus foliaceus and 1 pt. with epidermolysis bullosa acquisita (EBA). Patients were treated with a combination of protein A immunoadsorption (PAIA, 3–21 treatments) and rituximab (3–6 treatments) in addition to low dose conventional immunosuppression.

Results

All patients showed rapid clinical improvement starting within the first 4 weeks and decline of circulating autoantibody levels. Complete/partial remission was 88%/12% in pemphigus and 71%/29% in subepidermal blistering diseases. Overall relapse rate was 13% with an average follow‐up of 22 months. In the AL332‐MMP pt. the PAIA/rituximab treatment was stopped because of an oesophagus cancer considered as the paraneoplastic cause of the skin disease.

Conclusion

Combined treatment with PAIA and rituximab showed rapid and long‐lasting response, thereby allowing substantial reduction of dosage of concomitant immunosuppressive medication. We hereby confirm data from other investigators that PAIA/rituximab treatment is a promising therapeutical modality for pemphigus, pemphigoids and EBA, characterized by a favourable ratio of beneficial efficacy and minimized long‐term adverse effects.
Reasons for recall following conditional discharge: explanations given by male patients suffering from dual diagnosis in a London Forensic Unit
Authors: Chiringa J. , Robinson J. E. , Clancy C.
Published: May 2013

Accessible summary

Patients who have been discharged from forensic services often have conditions they have to abide by as part of their discharge, and failure to do so leads to recall. We interviewed six men who had been conditionally discharged from forensic services and then been recalled into hospital to find out what they thought went wrong.

The reasons they gave for why things went wrong included feeling that the system was unfair and made them feel like criminals even though they did not feel they had put anyone at risk. Some of them were not fully aware of the conditions they needed to adhere to, and some of them had breached the conditions but did not take responsibility for what had happened.

In addition, supervision was felt to be very controlling and disruptive rather than supportive when patients were often lonely, bored and needing support. Most participants reported that they experienced poor standards of aftercare in hostels they were required to reside in.

In the future, care of patients after conditional discharge should include better communication between patients and their supervisory team, recognition of the need for more support and improvements in the standards of care in hostels, as well as a collaborative approach to risk assessment that might reduce the frequency of relapse and readmission.

This study explores how male patients suffering from dual diagnosis in a forensic unit perceive being recalled and readmitted following conditional discharge and their views about how services might be improved. A qualitative approach was used drawing on grounded theory techniques. Audiotaped semistructured interviews collected data from a purposefully selected sample of six participants who had been recalled and met the inclusion criteria of the study. Data were analysed using the constant comparative method. Most participants perceived the recall system as unfair, inappropriately criminalized their behaviour and was based on an assessment of risk that they did not understand or accept. Participants were not fully aware of the conditions of their discharge, and most did not accept responsibility for their role in being recalled and blamed the system. Care following discharge was rarely seen as positive, and poor standards in hostels were reported by most participants. Supervision was often seen as disruptive and controlling, and focused more on surveillance rather than support. Better communication might have helped them understand and adhere to the conditions of their discharge. Participants identified the importance of family and friends to their recovery, the importance of having their own accommodation, and the need to be more independent.
A systematic review of parent role modeling of healthy eating and physical activity for their young African American children
Authors: Webber Kashica J. , Loescher Lois J.
Published: May 2013

Purpose

We systematically reviewed current literature on parent role modeling of healthy eating and physical activity as a strategy to influence healthy weight of middle school‐age African American children.

Conclusions

The evidence for parent role modeling is low to average. Most studies did not target African American participants or middle school‐age children.

Practice Implications

Nurses can begin to remedy deficient knowledge of parent role modeling by assessing parent attitudes, beliefs, and healthy behaviors, as well as the children's. Existing dietary and physical activity guidelines are not specific to African Americans but can be a starting point for educating parents.
Flotillins functionally organize the bacterial membrane
Authors: Bach Juri Niño , Bramkamp Marc
Published: May 2013

Proteins and lipids are heterogeneously distributed in biological membranes. The correct function of membrane proteins depends on spatiotemporal organization into defined membrane areas, called lipid domains or rafts. Lipid microdomains are therefore thought to assist compartmentalization of membranes. However, how lipid and protein assemblies are organized and whether proteins are actively involved in these processes remains poorly understood. We now have identified flotillins to be responsible for lateral segregation of defined membrane domains in the model organism Bacillus subtilis. We show that flotillins form large, dynamic assemblies that are able to influence membrane fluidity and prevent condensation of Laurdan stained membrane regions. Absence of flotillins in vivo leads to coalescence of distinct domains of high membrane order and, hence, loss of flotillins in the bacterial plasma‐membrane reduces membrane heterogeneity. We show that flotillins interact with various proteins involved in protein secretion, cell wall metabolism, transport and membrane‐related signalling processes. Importantly, maintenance of membrane heterogeneity is critical for vital cellular processes such as protein secretion.
Roles of two large serine recombinases in mobilizing the methicillin‐resistance cassette SCCmec
Authors: Misiura Agnieszka , Pigli Ying Z. , Boyle‐Vavra Susan , Daum Robert S. , Boocock Martin R. , Rice Phoebe A.
Published: May 2013

Methicillin‐resistant Staphylococcus aureus (MRSA) emerged via acquisition of a mobile element, staphylococcal cassette chromosome mec (SCCmec). Integration and excision of SCCmec is mediated by an unusual site‐specific recombination system. Most variants of SCCmec encode two recombinases, CcrA and CcrB, that belong to the large serine family. Since CcrA and CcrB are always found together, we sought to address their specific roles. We show here that CcrA and CcrB can carry out both excisive and integrative recombination in Escherichia coli in the absence of any host‐specific or SCCmec‐encoded cofactors. CcrA and CcrB are promiscuous in their substrate choice: they act on many non‐canonical pairs of recombination sites in addition to the canonical ones, which may explain tandem insertions into the SCCmec attachment site. Moreover, CcrB is always required, but CcrA is only required if one of the four half‐sites is present. Recombinational activity correlates with DNA binding: CcrA recognizes only that half‐site, which overlaps a conserved coding frame on the host chromosome. Therefore, we propose that CcrA serves as a specificity factor that emerged through modular evolution to enable recognition of a bacterial recombination site that is not an inverted repeat.
Improving population estimates by quantifying diving and surfacing patterns: A dugong example
Authors: Hagihara Rie , Jones Rhondda E. , Grech Alana , Lanyon Janet M. , Sheppard James K. , Marsh Helene
Published: May 2013

Diving animals are available for detection from above the water when environmental conditions are favorable and the animals are near the surface. The number of animals that are unavailable for detection needs to be estimated to obtain unbiased population estimates. The current availability correction factors used in aerial surveys for the dugong (Dugong dugon) allow for variation in environmental conditions but use the average time dugongs spend near the surface (i.e., constant availability corrections). To improve availability estimates, we examined location and dive data from nine dugongs fitted with satellite telemetry units and time‐depth recorders (TDRs) in eastern Australia. The effects of water depth, tidal conditions, and habitat types on dugong surfacing time were examined using generalized linear mixed models (GLMMs). We found that availability for detection differed with water depth, and depth‐specific availability estimates were often lower than the constant estimates. The habitat effect was less influential, and there was no tidal effect. The number of dugongs estimated using depth‐specific availabilities were higher than those obtained using constant availabilities across water depth. Hence, information on water depth can refine availability estimates and subsequent abundance estimates from dugong aerial surveys. The methodology may be applicable to other aquatic wildlife.
Qualitative study on the placement of Huntington disease patients in a psychiatric hospital: Perceptions of Maltese nurses
Authors: Scerri Josianne , Cassar Rebecca
Published: May 2013

Individuals with adult or juvenile Huntington disease can be cared for within psychiatric hospitals. In this paper, nurses' perceptions about the appropriateness of a psychiatric setting for these patients were explored. Semistructured interviews were conducted with 10 Maltese nurses involved in the care of these individuals. Their responses were analyzed using thematic analysis. Three main themes were identified from this study: (i) Huntington disease is not a mental illness; (ii) the lack of specialized staff and equipment within a psychiatric setting; and (iii) a need for alternative care options. The findings provide an insight into the perceptions of nurses, as they play a key role in the care and management of individuals with Huntington disease in a psychiatric setting. The findings demonstrated the need to provide alternative residential options in the community, and to improve the care and support provided both within psychiatric hospitals and the community through staff education and the provision of necessary facilities and equipment.
Tumor treating fields: a new frontier in cancer therapy
Authors: Davies Angela M. , Weinberg Uri , Palti Yoram
Published: May 2013

Tumor treating fields (TTFields) is a noninvasive, regional antimitotic treatment modality that has been approved for the treatment of recurrent glioblastoma by the U.S. FDA and has a CE mark in Europe. TTFields therapy delivers low‐intensity (1–3 V/cm), intermediate‐frequency (100–300 kHz), alternating electric fields to the tumor using transducer arrays placed on the skin around the region of the body containing the tumor. TTFields therapy affects metaphase, by disrupting mitotic spindle formation, and anaphase, by dielectrophoretic dislocation of intracellular constituents, resulting in apoptosis. TTFields therapy is frequency tuned to specific cancer cell types. The antimitotic effect of TTFields therapy has been demonstrated in multiple cell lines when the appropriate frequency was utilized. A phase III trial of TTFields monotherapy compared to active chemotherapy in recurrent glioblastoma patients established that TTFields therapy is associated with minimal toxicity, better quality of life, and comparable efficacy to chemotherapy. Ongoing and future trials will evaluate TTFields in newly diagnosed glioblastoma, solid tumor brain metastases, nonsmall cell lung cancer, and ovarian and pancreatic cancers.