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In this study, we present further experiences with the intravaginal culture (IVC) of oocytes and cleavage stages. IVC is a new technique for in-vitro fertilization, its principle consisting of fertilization of oocytes in an air-free plastic capsule which is placed into the maternal vagina. In a total of 45 patients, 22 were treated by IVC and 23 by a conventional in-vitro fertilization technique (IVF). The pregnancy rates for IVC and IVF amounted to 22.7% (5/22) and 17.4% (4/23), respectively. Our results indicate that IVC is a valuable new method for treatment of special cases of sterility. Furthermore, extracorporal stress factors such as light and low temperature can be minimized by IVC. We also consider the psychological factor for the patients as very important since the mother can actively participate in the early development of the conceptus. [ABSTRACT FROM PUBLISHER]

We present the first search for the pair production of dark particles X via K&#95;{L}^{0}→XX with X decaying into two photons using the data collected by the KOTO experiment. No signal was observed in the mass range of 40-110  MeV/c^{2} and 210-240  MeV/c^{2}. This sets upper limits on the branching fractions as B(K&#95;{L}^{0}→XX)<(1-4)×10^{-7} and B(K&#95;{L}^{0}→XX)<(1-2)×10^{-6} at the 90% confidence level for the two mass regions, respectively.

Administration of kainic acid induces acute seizures that result in the loss of neurons, gliosis and reorganization of mossy fiber pathways in the hippocampus resembling those observed in human temporal lobe epilepsy. Although these structural changes have been well characterized, the mechanisms underlying the degeneration of neurons following administration of kainic acid remain unclear. Since the lysosomal enzymes, cathepsins B and D, are known to be involved in the loss of neurons and clearance of degenerative materials in a variety of experimental conditions, we evaluated their potential roles in kainic acid-treated rats. In parallel, we also measured the levels and expression of insulin-like growth factor-II/mannose 6-phosphate (IGF-II/M6P) receptors, which mediate the intracellular trafficking of these enzymes, in kainic acid-treated rats. Our results showed that systemic administration of kainic acid evoked severe loss of neurons along with hypertrophy of astrocytes and microglia in the hippocampus of the adult rat brain. The levels and activity of cathepsins B and D increased with time in the hippocampus of kainic acid-treated rats compared to the saline-injected control animals. The expression of both cathepsins B and D, as evident by immunolabeling studies, was also markedly increased in activated astrocytes and microglia of the kainic acid-treated rats. Additionally, cytosolic levels of the cathepsins were enhanced along with cytochrome c and to some extent Bax in the hippocampus in kainic acid-treated rats. These changes were accompanied by appearance of cleaved caspase-3-positive neurons in the hippocampus of kainic acid-treated animals. The levels of IGF-II/M6P receptors, on the other hand, were not significantly altered, but these receptors were found to be present in a subset of reactive astrocytes following administration of kainic acid. These results, taken together, suggest that enhanced levels/expression and activity of lysosomal enzymes may have a role in the loss of neurons and/or clearance of degenerative materials observed in kainic acid-treated rats., (Copyright © 2014 IBRO. Published by Elsevier Ltd. All rights reserved.)

A new surgical technique for the treatment of vaginal aplasia is described. It consists of a modification of Vecchietti's method, allowing laparoscopic performance of the operation. 9 patients have been treated in this way since January, 1991 with good results. No complications were seen. Being virtually atraumatic, this method requires only brief hospitalisation, which is of vital importance in view of the psychological and physical problems this particular condition entails. Results are identical, so that this technique should always take precedence over the conventional Vecchietti method.

Different anaesthetic procedures that were used during an in vitro fertilisation and embryo transfer (IVF-ET) program have been analysed in order to determine their influence on plasma levels of estradiol, progesterone, prolactin, and beta-endorphin and results of IVF-ET. METHODS. Fifty-four patients awaiting transvaginal oocyte aspiration were randomised into three groups: (1) anaesthesia with ketamine as an induction agent and analgesic (n = 20); (2) general intubation anaesthesia using thiopentone for induction and enflurane for maintenance (n = 18); and (3) no anaesthesia (n = 16). Estradiol, progesterone, prolactin, and beta-endorphin were measured from day 3 to 14 referring to follicle aspiration. Differences between preoperative hormone levels and their intra- and postoperative peaks were analysed using the Kruskal-Wallis test (P < 0.03). The results were corrected using the Holms method (alpha = 0.05). RESULTS. No differences were observed in estradiol and progesterone levels (Figs. 1, 2). Prolactin levels were 1.4 times higher (P < 0.001) when ketamine was used and 2.2 times higher (P < 0.001) after short general anaesthesia than in the control group (Fig.3). Similar results were observed with respect to beta-endorphin: in comparison with the control group we found significant elevation by a factor of 2.1 when ketamine was used (P < 0.001). The discrepancy became even more marked with general anaesthesia: beta-endorphin was 3.9 times higher compared to the controls (P < 0.001) (Fig.4). Comparing the two groups who were given anaesthetics, prolactin and beta-endorphin levels were also significantly different (P < 0.001). The IVF procedure itself did not appear to be affected by different anaesthetic procedures during oocyte aspiration (Table 2). CONCLUSIONS. The increased prolactin and beta-endorphin plasma levels associated with ketamine and general anaesthesia reflect a significant alteration of the observed hormone levels. When anaesthesia is indicated, we try to avoid general intubation anaesthesia in favor of ketamine.

Lasers have been in use for 15 years in laparoscopic surgery. The CO2-laser in particular has become very popular in the field of gynaecology for the treatment of endometriosis and adhesions. Results achieved with the CO2-laser seem to be superior to those of conventional instruments, although only a small number of randomised prospective studies exist. Fibre lasers, such as the Nd:YAG-, KTP, and Argon-lasers combine vaporisation- and/or coagulation-effects within the same operating instrument. Time-consuming changes of instruments during the procedure are unnecessary. Nevertheless, technical problems and high costs of these laser systems on the one hand, and unfulfilled and unrealistic expectations by physicians and patients alike on the other hand hinder wide-spread use of this technique.

Different methods for follicular puncture which were used in an in vitro fertilization-embryo transfer program have been analyzed in order to determine their influence on plasma levels of estradiol, progesterone, prolactin and beta-endorphins. Thirty-eight patients awaiting oocyte aspiration were randomized into a laparoscopic (n = 20) and an ultrasound-guided oocyte aspiration group (n = 18). No differences were observed as far as estradiol and progesterone levels and embryo transfer rates were concerned. When using the laparoscopic technique, prolactin levels were found to be elevated 1.5 times in comparison with the ultrasound-guided aspiration procedure, which is highly significant (p < 0.001). Also, beta-endorphins peaked at a significantly higher level during laparoscopy (27.8 vs. 20.7 mol/l, p < 0.001). The enhanced prolactin and beta-endorphin levels reflect an increased stress response during laparoscopy. With respect to less trauma and shorter duration of the procedure, we now perform the sonographically guided transvaginal aspiration technique in our program almost exclusively.

A cytogenetic analysis could be performed on 343 out of 566 oocytes, which remained unfertilized or uncleaved during our IVF programme. The results were analysed with relation to the women's age by classifying the patients into two age groups (< or = 35 years and > 35 years). There was no significant difference for the corresponding total rates of abnormal oocytes (27.7% vs. 42.4%), the rates of aneuploidy (14.5% vs. 18.2%), and the rates of hyperhaploidy (4.8% vs. 3.0%). These results do not support the assumption of an increase in (numerically) abnormal gametes with increasing maternal age, which is of special interest in cases of trisomy 21. However, it is not possible to draw any definite conclusions, because of the low number of gametes, which were analysed in the upper age group.

A cytogenetic analysis could be performed on 343 out of 566 oocytes which remained unfertilized or uncleaved during our in vitro fertilization (IVF) programme. The results were analyzed in relation to the kind of ovarian stimulation by classification of the patients into two groups: treatment with human menopausal gonadotropin (hMG) or treatment with clomiphene citrate (CC). There was no significant difference for the total rates of abnormal oocytes (hMG: 28.6%, CC: 32.2%), the rates of aneuploidy (hMG: 14.4%, CC: 16.9%), and the rates of hyperhaploidy (hMG: 3.9%, CC: 8.5%). These results suggest that the kind of medication for induction of multiple follicular growth does not influence the rate of cytogenetic anomalies in human oocytes.

Human oocytes remaining unfertilized or uncleaved during in vitro fertilization (IVF) yield valuable information on the kind and frequency of numerical and structural chromosome aberrations after their fixation. This enables us to assess the maternal contribution to perturbations in the course of fertilization and early embryonic development. Therefore, 566 unfertilized or uncleaved oocytes from our IVF programme were processed for cytogenetic examination. 223 gametes were not analyzable. 260 oocytes were completely karyotyped and chromosome counts could be established for 83 cells. 243 eggs had the normal haploid chromosome complement whereas 100 cells (29.2%) were abnormal. In detail, we found the following aberrations: hypohaploidy = 10.2%, hyperhaploidy = 4.7%, diploidy = 9.0%, tetraploidy = 1.2%, structural anomalies = 2.6%, prematurely condensed sperm chromosomes = 3.2%. Six cells had to be considered in two of these categories. Our results and comparisons with other publications imply that the total rate of anomaies in human oocytes from stimulated cycles amounts to 20 to 30%. This high frequency might be one of the limiting factors for the success rate of IVF.

Between August 1986 and April 1988, 22 women with unruptured tubal pregnancy were treated by laparoscopy in the Department of Gynaecology and Obstetrics at the University of Ulm. Linear salpingotomy was performed using the contact Nd:YAG laser technique. The laser beam was delivered by special sapphire probes attached to a standard optical fibre for incision and coagulation. Two different shapes of sapphire probes were used. The specific laser properties and the application of vasopressin enabled treatment without any other incision instruments or coagulation agents. In one patient subsequent laparoscopy became necessary due to persisting trophoblastic tissue. Sixteen women were assessed subsequently by second-look laparoscopy or hysterosalpingography for tubal patency and possible formation of adhesions. In 14 (88%) women the tubes were patent, and in two women adhesions were seen.

In 45 women from an in vitro fertilization (IVF) program, the uterine and ovarian blood flows were investigated by vaginal Doppler sonography. The resistance index was used to evaluate the blood pattern. When comparing the patients who became pregnant after embryo transfer (ET [group I, n = 12]) with those who did not conceive (group II, n = 33), it is evident that in group I the vascular resistance of the uterine arteries is significantly lower on the day of follicular aspiration. No differences could be detected in the ovarian vessels. The data obtained so far suggest that the receptivity of the endometrium is a crucial factor for successful implantation. In the final analysis, this can be appraised not only on the basis of morphological but also of hemodynamic parameters.

The importance of periadnexal adhesions as cause in female infertility is widely recognized. The incidence of salpingo-ovariolysis (in the absence of tubal obstruction) has been reported as being 35% of the surgical procedures to promote fertility. In addition, one of the important causes of failure of reconstructive fertility surgery is related to postoperative adhesions (either reformed of formed de novo). Adhesions may encapsulate the ovary and/or the terminal end of the oviduct; they may disturb the physiologic spatial, relationship between the fimbriae and ovary and thus impair ovum pick up. Adhesions may also disturb ovarian function decreasing the number of ovulations. The importance of postoperative adhesions is evident also by the vast numbers of studies, published on this subject in the last decade.

Simultaneously with in vitro fertilization (IVF), the acridine orange (AO) test was performed on 51 patients according to the method of Tejada et al. This test allows a differentiation of spermatozoa with intact (double-stranded) and denatured (single-sanded) DNA. A significant relationship was demonstrated between the percentage of green-fluorescing (intact) sperm and the results of IVF. For both, the groups with (n = 40) and without (n = 11) fertilization of human eggs, a positive correlation was found between AO-test and sperm motility, and AO-test and normal sperm morphology.

The hamster ova penetration test (HOPT) was performed in 82 patients simultaneously with in vitro fertilization of homologous oocytes (IVF). A penetration rate of greater than 20% of inseminated ova was considered to represent a positive test result. The results of HOPT correlated completely with the IVF in all couples with male factor sterility. In couples with female sterility only and in couples with a combination of female and male factors HOPT was falsely negative in 22 and 20% respectively. Despite a negative HOPT in vitro fertilization with formation of two pronuclei 12 to 18 hours after insemination occurred in these patients. Obviously, the interpretation of HOPT is dependent upon the definition of the cut-off point (in our case 20%). Our results support the opinion that there should be no exclusion from an IVF programme based only on a negative HOPT. It appears that any penetration rate greater than 0% should be considered as a sign of potential fertility.