The importance of neuronal growth factors in the ovary.

The neurotrophin family consists of nerve growth factor (NGF), neurotrophin 3 (NT3) and neurotrophin 4/5 (NT4/5), in addition to brain-derived neurotrophic factor (BDNF) and the neuronal growth factors, glial cell line-derived neurotrophic factor (GDNF) and vasointestinal peptide (VIP). Although there are a few literature reviews, mainly of animal studies, on the importance of neurotrophins in the ovary, we aimed to provide a complete review of neurotrophins as well as neuronal growth factors and their important roles in normal and pathological processes in the ovary. Follicular assembly is probably stimulated by complementary effects of NGF, NT4/5 and BDNF and their receptors. The neurotrophins, GDNF and VIP and their receptors have all been identified in preantral and antral follicles of mammalian species, including humans. Transgenic mice with mutations in the genes encoding for Ngf, Nt4/5 and Bdnf and their tropomyosin-related kinase ß receptor showed a reduction in preantral follicles and an abnormal ovarian morphology, whereas NGF, NT3, GDNF and VIP increased the in vitro activation of primordial follicles in rats and goats. Additionally, NGF, NT3 and GDNF promoted follicular cell proliferation; NGF, BDNF and VIP were shown to be involved in ovulation; VIP inhibited follicular apoptosis; NT4/5, BDNF and GDNF promoted oocyte maturation and NGF, NT3 and VIP stimulated steroidogenesis. NGF may also exert a stimulatory effect in ovarian cancer and polycystic ovarian syndrome (PCOS). Low levels of NGF and BDNF in follicular fluid may be associated with diminished ovarian reserve and high levels with endometriosis. More knowledge of the roles of neuronal growth factors in the ovary has important implications for the development of new therapeutic drugs (such as anti-NGF agents) for ovarian cancer and PCOS as well as various infertility problems, warranting further research.

Cryopreservation of in vitro matured oocytes in addition to ovarian tissue freezing for fertility preservation in paediatric female cancer patients before and after cancer therapy.

STUDY QUESTION: Is a protocol that combines in vitro maturation of germinal vesicle-stage oocytes and their vitrification with freezing of cortical ovarian tissue feasible for use in fertility preservation for both chemotherapy-naive paediatric patients as well as patients after initiation of cancer therapy? SUMMARY ANSWER: Follicle-containing ovarian tissue as well as oocytes that can undergo maturation in vitro can be obtained from paediatric patients (including prepubertal girls) both before and after cancer therapy. WHAT IS KNOWN ALREADY: Anticancer therapy reduces the number of follicles/oocytes but this effect is less severe in young patients, particularly the paediatric age group. Autotransplantation of ovarian tissue has yielded to date 60 live births, including one from tissue that was cryostored in adolescence. However, it is assumed that autografting cryopreserved-thawed ovarian cortical tissue poses a risk of reseeding the malignancy. Immature oocytes can be collected from very young girls without hormonal stimulation and then matured in vitro and vitrified. We have previously shown that there is no difference in the number of ovarian cortical follicles between paediatric patients before and after chemotherapy. STUDY DESIGN, SIZE, DURATION: A prospective study was conducted in a cohort of 42 paediatric females with cancer (before and after therapy initiation) who underwent fertility preservation procedures in 2007-2014 at a single tertiary medical centre. PARTICIPANTS/MATERIALS, SETTING, METHODS: The study group included girls and adolescent females with cancer: 22 before and 20 after chemotherapy. Following partial or complete oophorectomy, immature oocytes were either aspirated manually ex vivo from visible small antral follicles or filtered from spent media. Oocytes were incubated in oocyte maturation medium, and those that matured at 24 or 48 h were vitrified. Ovarian cortical tissue was cut and prepared for slow-gradual cryopreservation. Anti-Mullerian hormone (AMH) levels were measured in serum before and after oophorectomy. MAIN RESULTS AND ROLE OF CHANCE: Ovarian tissue was successfully collected from 78.7% of the 42 patients. Oocytes were obtained from 20 patients before chemotherapy and 13 after chemotherapy. The youngest patients from whom oocytes were retrieved were aged 2 years (two atretic follicles) and 3 years. Of the 395 oocytes collected, ∼30% were atretic (29.6% in the pre-chemotherapy group, 37% in the post-chemotherapy group). One hundred twenty-one oocytes (31%) were matured in vitro and vitrified: 67.8% from patients before chemotherapy, the rest after chemotherapy. Mature oocytes suitable for vitrification were obtained from 16/20 patients before chemotherapy and from 12/13 patients after chemotherapy (maturation rate, 32 and 26.4%, respectively). There were significant correlations of the number of vitrified oocytes with patient age (more matured oocytes with older age) (P = 0.001) and with pre-oophorectomy AMH levels (P = 0.038 pre-chemotherapy group, P = 0.029 post-chemotherapy group). Oocytes suitable for vitrification were obtained both by manual aspiration of antral follicles (45%) and from rinse solutions after dissection. There were significantly more matured oocytes in the pre-chemotherapy group from aspiration than in the post-chemotherapy group after both aspiration (P < 0.033) and retrieval from rinsing fluids (P < 0.044). The number of pre-antral follicles per histological section did not differ in the pre- versus post-chemotherapy. AMH levels dropped by approximately 50% after ovarian removal in both groups, with a significant correlation between pre- and post-oophorectomy levels (P = 0.002 pre-chemotherapy group, P = 0.001 post-chemotherapy group). LIMITATIONS, REASONS FOR CAUTION: There were no patients between 5 years and 10 years old in the post-chemotherapy group, which might have affected some results and correlations. Oocytes from patients soon after chemotherapy might be damaged, and caution is advised when using them for fertility-restoration purposes. The viability, development capability and fertilization potential of oocytes from paediatric patients, especially prepubertal and after chemotherapy, are unknown, in particular oocytes recovered from the media after the tissue dissection step. WIDER IMPLICATIONS OF THE FINDINGS: Although more oocytes were collected and matured from chemotherapy-naïve paediatric patients, ovarian tissue and immature oocytes were also retrieved from young girls in whom cancer therapy has already been initiated. Our centre has established a protocol for potential maximal fertility preservation in paediatric female patients with cancer. Vitrified-in vitro-matured oocytes may serve as an important gamete source in paediatric female patients with cancer because the risk of reseeding the disease is avoided. Further studies are needed on the fertility-restoring potential of oocytes from paediatric and prepubertal patients, especially after exposure to chemotherapy. STUDY FUNDING/COMPETING INTERESTS: The study was conducted as part of the routine procedures for fertility preservation at our IVF unit. No funding outside of the IVF laboratory was received. Funding for the AMH measurements was obtained by a research grant from the Israel Science Foundation (to B.-A.I., ISF 13-1873). None of the authors have competing interests. TRIAL REGISTRATION NUMBER: N/A.

Optimizing the process of fertility preservation in pediatric female cancer patients - a multidisciplinary program.

BACKGROUND: Current evidence indicates sub-optimal incidence of fertility preservation (FP) in eligible patients. We present herein our designated multidisciplinary program for FP in pediatric and adolescent population and present our data on FP in female patients. METHODS: Pediatric patients (age 0-18) who were candidate for highly gonadotoxic treatments were referred to FP program for a multidisciplinary discussion and gonadal risk-assessment followed by either oocyte cryopreservation or ovarian cryopreservation (OCP) for female patients, and sperm banking for male patients. The OCP protocol consists of aspiration of oocytes from small antral follicles and in-vitro maturation followed by cryopreservation, as well as ovarian tissue cryopreservation. RESULTS: The establishment of a designated FP program resulted in a significant increase in referral and subsequent FP procedures of all eligible patients. Sixty-two female patients were referred for FP discussion during a period of 36 months; 41 underwent OCP; 11 underwent oocyte cryopreservation and six were declined due to parental decision. The median age was 13.2y (range 18 months-18y). Thirty-two (51.6 %) were chemotherapy-naïve. Seventeen patients (27 %) had sarcoma, 16 patients (26 %) had acute leukemia. The mean number of mature oocytes that were eventually vitrified was significantly higher in chemotherapy-naïve patients compared with chemotherapy-exposed patients (mean 12 oocytes (1-42) versus 2 (0-7)). CONCLUSION: Multidisciplinary programs that encompass experts of all relevant fields, skilled laboratory resources and a facilitated path appear to maximize the yield. We observed a considerable higher referral rates following launching a designated program and earlier OCP in chemo-naïve patients that culminated in a better fertility preservation procedure.

Keratinocyte growth factor and its receptor in human ovaries from fetuses, girls and women.

Keratinocyte growth factor (KGF) promotes growth of rat pre-antral follicles. There is limited information regarding its presence or that of its unique receptor (KGFR) in human ovaries, specifically in pre-antral follicles. The aim of the study was to investigate the expression of KGF and KGFR in ovarian samples from human fetuses and girls/women. The samples were prepared for immunohistochemical study of the KGF protein and for in situ hybridization to localize mRNA transcripts of KGFR. Total RNA was extracted from frozen ovarian samples, and the expression of KGF mRNA transcripts was investigated by reverse transcriptase polymerase chain reaction. In both fetuses and girls/women, the protein for KGF was detected from primordial stages in oocytes, granulosa cells (GCs) and stroma cells. Its mRNA transcripts were also detected in all extracts. The mRNA transcripts for KGFR were detected mainly in stroma cells in ovarian samples from both sources; in 10% of the samples, follicular staining was noted also in oocytes and GCs. Further studies adding KGF to the culture medium are needed to elucidate its putative role in human primordial follicle activation.

Platelet-derived growth factors (PDGF-A and -B) and their receptors in human fetal and adult ovaries.

There is no information regarding the presence of platelet-derived growth factors (PDGFs) and their receptors in human ovaries. The expression of PDGF-A, -B and their two receptors, PDGFR-alpha and -beta, was investigated in ovarian samples from women/girls and from human fetuses, at the protein and mRNA levels. The samples were prepared for immunohistochemical staining for PDGF-A and -B and their two receptors and in situ hybridization for the detection of the mRNA transcripts of the receptors. Total RNA was extracted from frozen ovarian samples, and the expression of PDGF-A and -B was investigated by reverse transcription-polymerase chain reaction. The proteins for PDGF-A and -B were detected in oocytes, and in granulosa cells (GC) of 50% of the follicles from women/girls. The proteins and mRNA transcripts for the two receptors were detected in oocytes (mRNA for PDGFR-beta only in 25% of the oocytes). PDGFR-alpha mRNA was expressed in GC of a minority of the samples from women/girls, whereas PDGFR-beta protein and mRNA were identified in over 50% of the GC from this source. PDGF-A and -B transcripts were identified in all the extracts. The presence of the receptors in GC suggests that PDGFs might be involved in the activation of primordial follicles.

Mental health symptoms in partial epilepsy.

We evaluated the potential clinical value of the Symptom Checklist-90-Revised (SCL-90-R) as a multidimensional self-report measure to identify the expected higher rates of clinically significant mental health symptoms in adults with partial/complex partial epilepsy (PE), as compared to a representative sample of adult non-patients. As expected, adults with PE had significantly higher rates of elevated SCL-90-R scale scores than did adult non-patients. The SCL-90-R may serve as both a screening measure to identify patients who could benefit from further mental health services as well as a measure of clinical response to epilepsy- and mental health-related interventions.

Ictal onset slow potential shifts recorded with hippocampal depth electrodes.

PURPOSE: Reports of direct current shifts at the onset of scalp-recorded seizures prompted us to inspect depth-recorded seizures for the presence of similar slow potential shifts at the onset of the seizure to determine whether slow potential (SP) shifts actually occur at the onset of depth-recorded seizures and if these shifts can facilitate localization of the seizure focus. METHODS: With the low frequency filter "opened" (LLF=0.1 Hz, HLF=70 Hz, 3 dB/octave), 32 seizures recorded with hippocampal depth and subdural electrodes were visually inspected to identify an SP shift at the onset of the seizure. A seizure was considered as having an SP shift when the slow potential waveform was > 1.5 sec in duration and > 100 microV in amplitude. Seizures were obtained from 5 subjects; 4 underwent epilepsy surgery (3=Engel I, 1=Engel II) and one received VNS. SP shift duration, peak voltage and polarity were measured for each seizure. The ability to identify seizures based on SP shift configuration was also evaluated. RESULTS: In 84% of the seizures, ictal onset was associated with a localized SP shift. Shift duration ranged from 1.5 sec to 11.5 sec (96% > 2 sec, 62% > 5 sec). The maximum shift ranged from 139 microV to 2305 microV (mean = 1123 microV, SD = 660 microV). In all the seizures, polarity was positive at the point of maximum shift. By visually examining the SP shift, seizures could be identified as originating from the same focus or from different foci. CONCLUSIONS: The onset of depth-recorded seizures appears to be commonly associated with a localized positive SP shift. An SP shift at the onset of depth-recorded seizures is likely to be a useful visual aid for localizing electrographic seizure onset.

Successful use of the Cryolock device for cryopreservation of scarce human ejaculate and testicular spermatozoa.

The existing methods for cryopreservation of very low count sperm samples are complex and sub-optimal for individual spermatozoa. Our purpose is to establish an effective simple method for cryoprotecting individual spermatozoa. Samples from patients with OTA were mixed with TYB or HEPES-buffered salt solution with glycerol + glucose and placed on a Cryolock that was plunged directly into liquid nitrogen or exposed to its vapors. Thawing was performed by direct immersion into a drop of warmed medium. The favorable method was tested on diluted samples (10-50 cells) and leftover TESE specimens from patients with azoospermia. Cryopreservation was considered successful if >30 spermatozoa, (>3 motile), or >5 spermatozoa (>1 motile) in diluted and TESE samples, were detected post-thawing. A significantly higher survival rate of seminal spermatozoa was obtained when using the Cryolock with TYB and freezing with liquid nitrogen vapor, compared to HEPES glycerol-glucose (95 vs. 35% respectively). Plunging the Cryolock into liquid nitrogen was detrimental. Cryolock combined with TYB cryoprotection and liquid nitrogen vapor freezing was highly effective for cryopreservation of individual spermatozoa in diluted and TESE samples. The Cryolock may serve for freezing very low-count sperm samples and individual spermatozoa. This method offers simplicity, efficacy, use of available materials, without requiring micromanipulation equipment or skills.

The effect of the hypoestrogenic state, induced by gonadotropin-releasing hormone agonist, on Doppler-derived parameters of aortic flow.

The effect of the hypoestrogenic state, induced by a GnRH agonist (GnRH-a), on cardiac function in healthy young women, was evaluated by Doppler echocardiography performed before treatment and when serum 17 beta-estradiol levels were suppressed by GnRH-a to 36.7 pmol/L. The following parameters of aortic flow were measured: peak flow velocity, ejection time, and acceleration time. Additional parameters calculated were flow velocity integral, cardiac index, and mean acceleration. The study group included 15 menstruating women, aged 25-42 yr (mean, 33 yr), with symptomatic fibroids, endometriosis, or scheduled for in vitro fertilization, who were treated with a GnRH-a. There were significant decreases in peak flow velocity (99 +/- 11 vs. 86 +/- 11 cm/s; P = 0.0004) and cardiac index (3.0 +/- 0.7 vs. 2.5 +/- 0.5 L/min.m2; P = 0.002). A decrease that did not reach statistical significance was noted in flow velocity integral (18.9 +/- 2.7 vs. 16.5 +/- 3.4 cm; P = 0.07). Mean acceleration was decreased significantly (12.6 +/- 2.6 vs. 10.8 +/- 1.8 m/s.s; P = 0.01), but no significant changes in acceleration time (81 +/- 16 vs. 83 +/- 10 ms; P = 0.7) or ejection time (296 +/- 25 vs. 295 +/- 27 ms; P = 0.8) were observed. These results indicate that estrogen deprivation is associated with smaller stroke volume and flow acceleration and might suggest that hypoestrogenism has a direct effect on cardiovascular performance.

The EEG response to diffuse and patterned photic stimulation during acute untreated alcohol withdrawal.

We studied the responses to diffuse and patterned stroboscopic light stimulation prospectively in 49 individuals during acute alcohol withdrawal prior to pharmacologic treatment. Photomyogenic responses (PMR) occurred in only two (4%) of those tested, and photoparoxysmal responses (PPR) never occurred. These findings suggest that PMR occur far less often during alcohol withdrawal than previously thought and that PPR may not be a direct manifestation of alcohol withdrawal.

Intraoperative radiation therapy for high-risk pediatric neuroblastoma.

PURPOSE: To evaluate the efficacy of intraoperative radiation therapy (IORT) in the treatment of high-risk pediatric neuroblastoma. METHODS AND MATERIALS: Between 1986 and 1998, 23 children received IORT for pediatric neuroblastoma. Electron beam energies ranged from 4 MeV to 16 MeV and median dose was 10 Gy (7-16 Gy). RESULTS: Twenty-one of 23 patients were classified as high-risk. A gross total resection (GTR) was achieved in 18 patients, of whom 6 experienced disease recurrence, 2 of which included a locoregional relapse as a component of failure. Fourteen of 18 patients receiving IORT after a GTR are disease-free survivors. A second subset of 5 patients had a subtotal resection (STR), with gross residual disease remaining after surgery. All 5 patients recurred locally, and all died of their disease. IORT was extremely well-tolerated in our cohort. Surgical resection and IORT resulted in the narrowing of the abdominal aorta and an atrophic kidney in 1 patient. CONCLUSIONS: For high-risk neuroblastoma patients, IORT as the only radiotherapy to the primary, produced excellent local control after a GTR. However, IORT as the sole radiotherapy to the primary was inadequate for patients with extensive adenopathy or an STR. In this setting, we are exploring the use of IORT as a boost in conjunction with external beam radiation therapy.

Cellular basis of luteal steroidogenesis in the human ovary.

A primary monolayer cell culture system was developed to investigate human corpus luteum (CL) function in vitro. Steroidogenic cells were isolated by collagenase dispersal and Percoll density-gradient fractionation from CLs enucleated at progressive stages of the luteal phase (tubal surgery patients). 'Pure' granulosa-lutein cells were aspirated from ovulatory follicles at mid-cycle (in-vitro fertilization patients). The steroidogenic capacity (progesterone/20 alpha-dihydroprogesterone biosynthesis and aromatase activity) of isolated luteal cells was assessed in relation to CL development. Basal luteal cell steroidogenesis was maximal at around the expected time of ovulation and declined with CL age during the luteal phase. Conversely, human chorionic gonadotrophin (hCG)-responsive steroidogenesis was initially undetectable but developed as the luteal phase progressed. These results show that luteal cell steroidogenesis becomes increasingly dependent upon gonadotrophic support with CL age. This is evidence that functional luteolysis in human ovaries (1) is pre-programmed to occur at the cellular level, (2) is initiated automatically at the time of ovulation and (3) is reversed at the time of CL 'rescue' in early pregnancy by the direct action of trophoblastic hCG on steroidogenic luteal cells. The culture system described should be of value in further defining the control of human CL form and function at the cellular level.

Placental isoferritin as a marker of early abortion in pregnancies induced by in vitro fertilization.

The relatively high rate of early pregnancy loss in artificial reproductive technology-induced conceptions has driven researchers to seek for an efficient diagnostic tool for estimating the gestational risk in these cases. Monitoring early gestation normalcy using serial beta-human chorionic gonadotrophin (beta hCG) measurements requires several days before diagnosis can be established. The objective of this study was to determine whether placental isoferritin (PLF) can be used as a predictive marker of normal pregnancy development during early stages of in vitro fertilization (IVF)-induced pregnancy. Ninety-three consecutive IVF cycles were investigated. Blood samples for PLF (enzyme linked immunosorbent assay; ELISA) and beta hCG (radio-immunoassay; RIA) determination were obtained from all women on days 11, 13 and 15 following embryo transfer. Placental isoferritin was detectable in the serum 11 days after embryo transfer in IVF conception cycles. These levels were significantly higher in normally developing pregnancies (n = 18) than in cases which eventually aborted spontaneously (n = 9) during the first trimester (mean +/- s.d.; 33 +/- 28 U/mL as compared with 1 +/- 2 U/mL; P = 0.0004; Wilcoxon test; sensitivity 94.5 per cent, specificity 88.9 per cent, positive predictive value 89.9 per cent, negative predictive value 94.5 per cent). Moreover, in those patients who later aborted, lower than normal PLF levels were detected long before the decline in beta hCG titres was evident. Considering its suppressor activity, it is expected that PLF levels would be high at the initiation of normal pregnancy. This may explain the present finding of low PLF levels in abnormally developing IVF-induced pregnancies. These preliminary data suggest that PLF can be used as a sensitive marker for detecting those cases destined to abort at a very early stage. However, further studies are still required on spontaneous conceptions, before this test can be recommended for routine clinical application.

Membranous dysmenorrhea: the forgotten entity.

BACKGROUND: Membranous dysmenorrhea involves the spontaneous slough of the endometrium in one cylindrical or membranous piece that retains the shape of the uterine cavity. Because this entity is rarely mentioned in the medical literature, the purpose of this report is to describe two such cases. CASES: An 18-year-old nullipara with regular menstrual cycles presented with membranous dysmenorrhea after taking the contraceptive Gynera (Gestodene 0.075 mg, ethinyl estradiol 0.030 mg). Symptoms disappeared when the medication was discontinued. The second patient, a 26-year-old woman, gravida 1, para 1, was on a 10-day monthly regimen of Provera (medroxyprogesterone acetate) 2.5 mg/day for dysfunctional uterine bleeding. When the Provera dose was increased to 10 mg/day, the symptoms disappeared. CONCLUSION: Membranous dysmenorrhea is "a disease of theories" with various recommended medications. When this condition is caused by iatrogenic treatment, the best approach is to discontinue the offending drug or change its dosage.

Abdominal pregnancy following in vitro fertilization in a patient with previous bilateral salpingectomy.

BACKGROUND: We present the first case of abdominal pregnancy after in vitro fertilization and embryo transfer in a patient without oviducts. CASE: A 38-year-old woman, who previously had had two salpingectomies because of two tubal pregnancies, was admitted to our department with intermittent vaginal bleeding and abdominal pain, 21 days after embryo transfer. Exploratory laparotomy revealed a moderate amount of blood in the peritoneal cavity and a mass consisting of blood clots and tissue fragments attached to the posterior aspect of the right broad ligament. Pathologic examination confirmed the diagnosis of abdominal pregnancy. CONCLUSION: Abdominal pregnancy may be the outcome of embryo transfer and should hence be considered a potential complication of the procedure.

Cutaneous graft-versus-host-like reaction after paternal lymphocyte immunization for prevention of recurrent abortion.

We have presented an immunocompetent patient who, after paternal lymphocyte immunization, developed a GVH-like skin reaction; she recovered spontaneously. This complication is rarely seen. This was the only case observed in our series of over 70 patients immunized. A similar phenomenon was not reported in a larger series by Carp and colleagues (1). However, the case deserves special attention because this reaction can be life-threatening. Therefore, we suggest that paternal lymphocyte immunization should be recommended only for those patients who have undergone a thorough work-up and in whom all causes, other than immunological, for recurrent abortion have been excluded. Even then, this treatment should be given with utmost caution. In view of the above described complication, we believe that more data are still required to justify offering such a treatment modality on a routine basis.

Human gamete fusion test: an experimental model for assessing the fusion potential of sperm from nonfertilizing normospermic patients.

OBJECTIVE: To develop an experimental model for assessing the ability of sperm to bind and subsequently fuse with the oolema and to use this test for evaluating the fusion potential of nonfertilizing human sperm. DESIGN: Aged human oocytes (n = 108) were denuded of their zonae and loaded with DNA-specific bisbenzamidazole fluorochrome. Sibling oocytes were inseminated by semen samples from either normospermic patients (study group; n = 12) who have repeatedly failed to achieve fertilization in vitro or patients demonstrating high fertilization rates (control; n = 12). Gamete fusion was ascertained by fluorescent microscopy and validated by scanning electron microscopy. Test results were analyzed in relation to current partial zona dissection treatment outcome. SETTING: Infertility and IVF Unit of an academic tertiary referral medical center and university-based basic research laboratory. RESULTS: In samples achieving fertilization by partial zona dissection (7/12; 9% to 23% fertilization rate), the mean numbers of fused spermatozoa were similar to those of the control (2.1 compared with 2.6) and significantly higher than in the partial zona dissection-failed fertilization subgroup. In six of seven cases in which partial zona dissection yielded embryos, fluorescent spermatozoa were detected on the oolema, whereas in all but one partial zona dissection-failed cases gamete fusion was not observed. CONCLUSIONS: The suggested human gamete fusion test may help focus on specific sperm dysfunction in nonfertilizing patients. It correlates well with the outcome of partial zona dissection-assisted IVF trials and may provide useful information before this treatment is attempted.

Tissue effects of a new laparoscopic carbon dioxide laser probe.

The effects of CO2 laser beam delivered through a new laparoscopic probe on rabbit uteri is presented. This device has an outer diameter of 4.9 mm and can be delivered through most of the standard operative laparoscopic accessories. Two probes with miniature zinc selenide lenses of 6.5 mm and 17 mm back focal length attached at the distal end were tested. Through each probe, constant 0.5-second pulses and power levels of 5, 10, and 15 W were applied in three different lens-to-target distances. Measurements of beam spot diameters and microscopic crater characteristics revealed significant differences as a result of minor shifting of lens-target distance. From 7 to 9 mm beyond the focal plane and at power levels of 5 to 10 W, negligible tissue effects could be detected. This was defined as an "optical back-stop" effect. A new application mode using an easily maneuverable probe for cutting and coagulating [focusing/defocusing] effects increases the options available for operative laparoscopy.

Detection of immunoglobulin A anticardiolipin antibodies in cervical mucus from in vitro fertilization patients and fertile women.

OBJECTIVE: To identify and quantify antiphospholipid autoantibodies of the immunoglobulin (Ig) A isotype in cervical mucus obtained from IVF patients and fertile controls. DESIGN: The study was performed prospectively. Blood and cervical mucus samples were obtained from patients undergoing IVF treatment (n = 27) at the time of expected E2 peak, before administering hCG. Control samples were taken from fertile women (n = 16) around the time of ovulation during a spontaneous nonstimulated menstrual cycle. Anticardiolipin activity was tested using ELISA. SETTING: Infertility and IVF unit of an academic tertiary referral medical center and university-based basic research laboratory. RESULTS: Forty-eight percent (13/27) of the IVF patients and 43.8% (7/16) of the fertile controls exhibited anticardiolipin IgA activity in aspirated cervical mucus. The mean activity measured for the positive cases was similar in both groups. This activity was higher than that measured in peripheral blood of the women studied. No difference was noted between infertile patients undergoing IVF treatment and fertile women in this respect. CONCLUSIONS: In this preliminary work, we demonstrated for the first time anticardiolipin IgA activity in human cervical mucus. These observations have to be substantiated by larger scaled studies to assess their possible clinical significance.