In vitro induction of cytotoxic effector cells against human neoplasms. I. Sensitization conditions and effect of cryopreservation on the induction and expression of cytotoxic responses to allogeneic leukemia cells.

Peripheral blood lymphocytes (PBL) from normal human donors were sensitized in vitro against allogeneic human acute myelocytic leukemia (AML) cells by means of an unidirectional mixed lymphocyte-tumor cell culture (MLTC) technique. The cytotoxic responsiveness of the sensitized lymphocytes, as determined in vitro by the 51Cr-release assay, varied among individual lymphocyte donors and was greatly dependent on the sensitization culture conditions. Induction of cytotoxic effector cells was augmented appreciably by adding to the cultures minute amounts of the immunopotentiating agent MER-BCG. Responding lymphocytes and stimulating leukemia cells cryopreserved for several weeks in liquid nitrogen were as effective as fresh cells in generating effector lymphocytes; the cytotoxic capacity of already sensitized lymphocytes was fully retained by cryopreservation. The implications of these findings for possible clinical employment of in vitro sensitized lymphocytes in adoptive immunotherapy of cancer are discussed.

Second-look operative laparoscopy 1 year following reproductive surgery.

To evaluate the efficacy of a second-look operative laparoscopy following a reproductive surgery, a randomized study was conducted on women who failed to conceive 1 year following terminal salpingostomy or following salpingo-ovariolysis by laparotomy. There is no significant difference in the cumulative probability of pregnancy at 12, 24, and 36 months follow-up between women who underwent second-look operative laparoscopy 1 year after salpingo-ovariolysis (27%, 67%, and 67%) and women who were continued to be observed expectantly (27%, 45%, and 52%, respectively). The cumulative probability of conception at 12, 24, and 36 months follow-up in women who underwent second-look laparoscopy 1 year after salpingostomy was 6%, 18%, and 18% and in women who were observed expectantly was 5%, 21%, and 27%, respectively (P = no significant). The incidence of ectopic pregnancy which was high (about one-half of patients who conceived) was also not affected by this procedure. This study suggests that second-look operative laparoscopy 1 year after terminal salpingostomy or salpingo-ovariolysis does not increase the pregnancy rate or decrease the incidence of ectopic pregnancy. Women who fail to conceive 1 year after a reproductive surgery still maintain some of their reproductive potential, but the risk of having an ectopic pregnancy is high.

Pregnancy rates after communal growth of preimplantation human embryos in vitro.

OBJECTIVE: To evaluate the effect of communal growth of preimplantation human embryos on pregnancy rates in IVF-ET. DESIGN: Prospective randomized study. SETTING: In Vitro Fertilization Unit, Bikur Cholim Hospital, Jerusalem, Israel. PATIENTS: Ninety-one women undergoing 91 cycles of IVF-ET. INTERVENTION: In 42 cycles (control) embryos were grown singly, in 49 cycles (study) embryos were grown in groups until transfer. MAIN OUTCOME MEASURES: Proportion of women achieving pregnancy in study and control groups. RESULTS: Communal growth of embryos resulted in significantly improved pregnancy rates. Intrauterine transfer of embryos grown in groups produced 43% pregnancy rates per transfer, whereas, with embryos grown individually, 24% pregnancy rates were obtained (odds ratio = 2.4). CONCLUSIONS: Our study suggests that, in IVF, the potential of preimplantation human embryos to produce pregnancy tends to increase when embryos are grown in groups.

Electroejaculation in combination with intracytoplasmic sperm injection in patients with psychogenic anejaculation results in lower fertilization rates.

OBJECTIVE: To evaluate the outcome of intracytoplasmic sperm injection (ICSI) with sperm obtained by electroejaculation in men with psychogenic anejaculation. DESIGN: Retrospective clinical study. SETTING: In Vitro Fertilization Unit, Bikur Cholim Hospital, Jerusalem, Israel. PATIENT(S): Seven men with psychogenic anejaculation who underwent 16 sessions of electroejaculation in combination with ICSI. INTERVENTION(S): Electroejaculation, ICSI. MAIN OUTCOME MEASURE(S): Semen analysis, ICSI, fertilization rates. RESULT(S): All patients had poor sperm motility. One hundred forty-seven oocytes were injected, with a fertilization rate of 27% (39/142). One ongoing pregnancy was achieved. CONCLUSION(S): Sperm obtained by electroejaculation have low motility and reduced fertilization potential. Nevertheless, ICSI should be offered to improve the possibility of successful pregnancy.

Soluble ICAM-1 and E-selectin levels correlate with clinical and biological aspects of severe ovarian hyperstimulation syndrome.

OBJECTIVE: To assess the potential involvement of the soluble endothelial cell-leukocyte adhesion molecules E-selectin and intercellular adhesion molecule-1 (ICAM-1) in the pathophysiology of capillary hyperpermeability in the ovarian hyperstimulation syndrome (OHSS). DESIGN: Controlled clinical study. SETTING: Women hospitalized for severe OHSS after ovulation induction for IVF at two tertiary medical centers. PATIENT(S): Eleven patients with severe OHSS undergoing IVF and 20 controls who had received a similar ovulation induction regimen and did not develop OHSS. INTERVENTION(S): Serial serum samples were obtained from all patients with OHSS from admission until discharge. Ascitic fluid was obtained from all patients by therapeutic paracentesis. Serum was obtained from all controls 4-8 days after ET. MAIN OUTCOME MEASURE(S): Samples were assayed for soluble E-selectin and soluble ICAM-1 by using enzyme-linked immunosorbent assay and results were correlated with clinical and biological aspects of OHSS. RESULT(S): Compared with controls, patients with severe OHSS had higher levels of soluble ICAM-1 and lower levels of soluble E-selectin detected in serum and ascites. Serum levels of soluble ICAM-1 decreased, while soluble E-selectin levels increased along with clinical and biological improvement. Serum soluble ICAM-1 showed significant positive correlation and serum soluble E-selectin showed significant negative correlation with clinical and biological aspects of severe OHSS. CONCLUSION(S): Soluble ICAM-1 and soluble E-selectin seem to be involved in the pathophysiology of capillary hyperpermeability in severe OHSS.

Baby bottle tooth decay and complications during pregnancy and delivery.

The purpose of the study was to investigate an association between maternal and/or fetal complications during pregnancy and/or delivery and the occurrence of baby bottle tooth decay (BBTD) in the infant. The study population comprised 50 mothers of infants with BBTD (BBTD+ mothers): 50 mothers of aged-matched children with similar eating and bottle-use habits but without BBTD (BBTD-) served as comparisons. Interviews with the mothers focused on pregnancy complications (vaginal bleeding, premature uterine contractions, viral or bacterial infections, hospitalization, diabetes, or other causes of high-risk pregnancy), mode of delivery (normal or instrumental), fetal distress, gestational age, birth weight, and mother's age at delivery. Chi-square analysis and the Student's t-test were used to analyze the data. Pregnancy complications and instrument delivery/cesarean section were significantly higher in the BBTD+ group than in the controls (P = 0.0001 and P = 0.0004, respectively). In the BBTD+ group, vaginal bleeding/premature uterine contractions were most frequent (50%), while in the BBTD- group high-risk pregnancy dominated (20%). Normal deliveries were more frequent in the BBTD+ mothers (68%) than in the comparisons (40%). Gestational age and baby's birth weight did not differ between the two groups. The mean age of the BBTD+ mothers was significantly lower than that of the controls (P = 0.013). The results suggest that babies born after maternal complications during pregnancy or babies who experience a traumatic birth must be considered to be at risk of developing BBTD when exposed to excessive bottle nursing.

Antisperm antibodies in men with psychogenic anejaculation.

Antisperm autoantibodies were determined in 16 men suffering from psychogenic anejaculation who underwent assisted reproduction treatments. Blood and semen samples were collected after transrectal electroejaculation and antisperm antibodies in serum and on the surface of motile spermatozoa were measured using the direct and indirect immunobead binding test. Five men (31%) were found positive for antisperm antibodies. The majority of antibodies were directed against the sperm heads. Surface antibodies were mainly IgA isotype whereas serum antibodies were IgG isotype. These results suggest that psychogenic anejaculation might be associated with increased incidence of antisperm autoimmunity.

[Initial results of a cryopreservation program for human ova and embryos]. / Erste Ergebnisse eines Kryokonservierungsprogrammes menschlicher Eizellen und Embryonen.

A human egg and embryo freezing program for non fertilized eggs and spare embryos was established recently in our ib vitro fertilization clinic. The first results are reported. Two cryoprotectants, glycerol and dimethylsulfoxid (DMSO) were used in an automatic computerized freezing program. DMSO gave us better results in terms of a 55% survival of all blastomeres (from 6 normal and 5 pathologic 2- to 16-cell embryos) after thawing. Some of the thawed embryos continued cleavage in vitro. Four embryo replacements were performed with the 6 normal embryos, but none of them resulted in pregnancy. The rapid technique using glycerol gave us less good results in terms of blastomere survival (34%); however, from 3 embryo replacements one "biochemical pregnancy" was obtained from a thawed four-cell embryo. At first attempts to freeze and thaw unfertilized eggs, DMSO seemed also to give better results. The presented cryopreservation techniques which are modifications of those published by Trounson et al. and Zeilmaker et al. need further improvement although the first results are encouraging.

Incidence of post-abortion intra-uterine adhesions evaluated by hysteroscopy--a prospective study.

A prospective study was conducted on the incidence of intra-uterine pathology diagnosed by hysteroscopy in 147 women who underwent dilatation and sharp curettage due to spontaneous first trimester abortion. The diagnostic hysteroscopies were performed 4-6 weeks after the miscarriage, using CO2 as medium for uterine distention. In 38 patients (25.9%), uterine pathology was found, of whom 28 patients (19%) had intra-uterine adhesions (IUA). The incidence of IUA following only one abortion was 16.3% (16 out of 98 cases), all were of mild extent and filmy consistency, occupying less than one-quarter of the uterine cavity. The incidence of IUA after two abortions was 14% (3/21) but the incidence after three or more spontaneous abortions was significantly elevated (32%, 9/28), (P < 0.05, chi-square test). In addition, 58% of IUA diagnosed in the latter two groups were of a more severe extent. After a first miscarriage, occurrence of post-abortion IUA was not related to the serum oestrogen concentration on the day of curettage, nor was it related to any of the oestrogen treatment modalities used in this study. Although in most of these cases normal recovery of the endometrium was the rule, the high incidence of IUA diagnosed in this prospective study stresses the need for preventive treatment and close follow-up of patients after sharp curettage due to spontaneous abortion. Obviously oestrogen treatment alone did not prove to be effective and other treatment methods should be evaluated. The high incidence and severity of IUA in habitual aborters clearly justifies the performance of a diagnostic hysteroscopy as a standard evaluation.

Immunity to zona pellucida in women with low response to ovarian stimulation, in unexplained infertility and after multiple IVF attempts.

A retrospective study was designed to assess the presence of antizona pellucida autoantibodies in in-vitro fertilization (IVF) patients in relation to low ovarian response, multiple IVF attempts and unexplained infertility. Antizona pellucida and antisperm antibodies were determined in serum samples obtained from 37 women undergoing IVF-embryo transfer and 20 fertile women. Antizona pellucida antibodies were measured using enzyme immunoabsorbent assay. Antisperm antibodies were evaluated by the immunobead binding method. Three of 10 patients with low response to ovarian stimulation had antizona pellucida antibodies in serum and one patient had antisperm antibodies. None of the other participants in the study and the control groups demonstrated measurable levels of serum antigamete antibodies. The results in this small group suggest an association between antizona pellucida antibodies and suboptimal response to gonadotrophins. It is indicated that repeated stimulation and puncture of ovaries in IVF procedures do not elicit autoimmunity to gametes.