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In an IVF-program a total of 585 oocytes (180 patients) was examined for the presence of pronuclei 16 to 20 hours after the addition of spermatozoa. The fertilization rate was 71% and in 58 (10%) of the fertilized oocytes three or more pronuclei, indicating a failure of the block to polyspermy with polyspermic fertilization, could be observed. The frequency of polyspermy was related to the maturity of the oocyte, determined according to morphological criteria. Immature oocytes showed a higher percentage of polyspermic fertilization (32%) compared to mature oocytes (6%). Preincubation of oocytes (0-1.5, 2-4, 5-8 hours) prior to the addition of spermatozoa increased the fertilization rate (67%, 70%, 83%, respectively). The polyspermy rate seemed to increase (13%, 14%, 19%). The observed frequency, however, was not significantly different between the various preincubation intervals. The polyspermy rate was affected by the number of spermatozoa used for in-vitro fertilization. Insemination with 0.5-0.8, 1.0 or 1.5 X 10(6) spermatozoa/oocyte resulted in a polyspermy rate of 6%, 20%, 32%, respectively. The appearance of polyspermic fertilization was not related to the age of the patient (20 to 45 years) nor to the method of ovarian stimulation (Clomiphene, hMG, Clomiphene/hMG). Because of the high incidence of polyspermy under in-vitro conditions it seems to be important to examine the oocytes in the pronuclear stage on a routine basis. Reduction of the number of spermatozoa used for in-vitro fertilization and the exact timing of insemination according to the maturity of the oocyte might reduce the occurrence of polyspermic fertilization.

11 patients with advanced, recurrent mamma cancer were treated by a PAC polychemotherapy. The therapy was well tolerated, the incidence of side effects was low. All patients responded to therapy (7 NED, 3 CR, 1 PR). 2 patients died, one commited suicide, the other died after having developed hepatitis. Of the remaining 9 patients 7 survive recurrence free; medium follow up 20 months. These data are in accordance with results obtained by Kolarić et al. [2] and Forastiere et al. [1] who showed platinum to be effective in the treatment of mamma cancer.

Between December 1986 and November 1987, 588 transvaginal sonographically guided oocyte retrievals using a vaginal transducer were performed for in-vitro fertilization. All follicles were accessible. No complication occurred except for one case of pelviperitonitis in a patient with preoperatively diagnosed sactosalpinx. Because this technique is safe, non-invasive and can be performed without general anaesthesia, eggs were also retrieved from patients despite poor hormonal values and endogenous LH surge. In all, clinical pregnancies were diagnosed in 85 patients. Pregnancy rates per embryo transfer, oocyte retrieval and stimulated cycle, respectively, were 19.0, 14.5 and 12.0%. The advantages of the transvaginal technique compared with laparoscopically guided and with other sonographically guided techniques of oocyte retrieval are obvious, e.g. no general anaesthesia, in general all follicles accessible, very low complication rates, little discomfort to patient, less time and fewer staff. Therefore, this technique seems to be the method of choice. Laparoscopically guided oocyte retrieval is only indicated when a laparoscopic screening of pelvic organs or a gamete intra-Fallopian transfer are to be performed simultaneously.

The number and viability of spermatozoa recovered by glass wool column filtration and a swim-up procedure were compared using different types of ejaculates, such as normal, asthenozoospermic and very viscous oligozoospermic semen. The filtration procedure resulted in significantly (P less than 0.01) higher recovery of viable spermatozoa than the swim-up procedure from all types of ejaculates studied. Further, the spermatozoa from 50 (78.1%) of the 64 ejaculates filtered through glass wool column fertilized at least one intact human egg in an in-vitro fertilization (IVF) procedure. It is concluded that glass wool column filtration is superior to the swim-up procedure since it yields a higher recovery of viable spermatozoa that are potentially fertile. Therefore, the glass wool column filtration procedure used to prepare spermatozoa may be of benefit for IVF, intra-uterine insemination, in-vitro fertilization and GIFT (gamete intra-Fallopian transfer), especially in cases of poor quality semen.

The therapeutic efficacy of stimulating ovarian follicular development by pure urinary FSH in an IVF programme was determined and compared with results of other therapeutic regimens. Forty-three patients selected for extracorporeal fertilization were treated with pure FSH after stimulation with clomiphene or HMG had proved to be unsuccessful. Treatment had to be discontinued in five patients before follicular puncture, and embryo transfer was performed in 32, of whom seven conceived. FSH stimulation evidently creates favourable conditions for in-vitro fertilization in patients with impaired follicular maturation following conventional stimulation with clomiphene or HMG. Levels of oestradiol, progesterone and androstenedione were measured in follicular fluid, and high progesterone concentrations were correlated with mature oocytes. The increased ratios of progesterone/androstenedione and progesterone/oestradiol in follicles containing mature oocytes are due to the onset of luteinization of mature follicles as a consequence of HCG administration.

Ejaculates obtained from men attending an in vitro fertilization (IVF) program were analyzed to determine the effect of the presence of leucocytes in semen on the ability of sperm in that semen to fertilize oocytes. The mean (+/- S.E.) concentration of leucocytes in 103 ejaculates was 0.62 X 10(6)/ml (+/- 0.09) with a range of 0.1 X 10(6) or less per ml to 7.0 X 10(6)/ml. The leucocyte concentration demonstrated a significant (p less than 0.05) but inverse relationship with the fertilizing capacity of spermatozoa. Though not statistically significant, the leucocyte concentration also demonstrated an inverse relationship to sperm motility and normal morphology. These results suggest that the presence of leucocytes in an ejaculate cannot be used as the sole indicator of the fertilizing capacity of spermatozoa in that ejaculate.

Passive electrical properties of oocytes and of zonae pellucidae, and the mechanical coupling between them, can be elucidated by means of rotating-field-induced rotation. In low-conductivity media (25-100 microS/cm) rotation of mouse oocytes (with or without their zonae) requires fields in the 1-100 kHz frequency range. However, an isolated zona shows weak rotation in the opposite direction to that of a cell, and in response to much higher field frequencies (approx. 1 MHz). In zona-intact mouse oocytes, the rotation of cell and zona are not rigidly coupled: thus rotation of the cell can still be induced when the zona is held stationary. However, rotation of freely suspended zona-intact cells is much slower than that of zona-free cells and requires an optimum field frequency that is approximately 1.5 kHz higher. These observations show that the electrical properties of the oocyte that are measured by rotation are altered by the presence of the zona pellucida, even though no such influence has been detected using micro-electrodes. The data are consistent with the zona acting as a porous shell with a conductivity of 40 microS/cm (preliminary estimate made at a single medium conductivity of 26 microS/cm). Measurements on cells from which the zonae had been removed gave values for the membrane capacity and resistivity of 1.2-1.3 microF/cm2 and 400 omega.cm2, respectively. These values may reflect the presence of plasmalemma microvilli. The results strongly suggest that the technique may be useful for studies of cell maturation and for in vitro fertilization, because the cells may be further cultured after measurement.

Unlabelled: Tissue slides of 230 epithelial ovarian cancers were examined by two different pathologic teams to determine the following details: histological pattern, grade of polymorphy, mitotic index, density of chromatin, presence of nuclei and macronuclei, lymphocytic infiltration, psammoma bodies, and necrosis. For statistical evaluations the data were presented together with stage, age and survival times in form of an information vector for each of the 230 patients and analysed using hazard-models (Cox 1972, Carter 1983) and contingency tables., Results: The agreement between the 2 pathologic teams was good. There were many interrelated histological concomitants, such as polymorphy and chromatin density, and as nuclei and macronuclei. Necrosis was relatively independent. Significant effects on prognosis could be demonstrated for polymorphy, chromatin density, mitotic index, histologic pattern, (macro)nuclei. Polymorphy was the best histological prognostic indicator and, together with necrosis, produced the best prognostic grading system.

In 25 patients with cervical, uterine and ovarian cancer with a depletion of T-helper cells and a lack of reactivity in the LMI-Test a course of immunotherapy with thymopentin (Timunox, Cilag Biotech) at a dosage of 100 mg/die for 2 weeks and subsequently 100 mg 3 x/week for 4 weeks was performed and a comparison was made with a control group of 59 patients receiving no immunotherapy. The following parameters of cell-mediated immunity were determined: (1) leucocyte-migration-inhibition-test (LMI-test) against the tumor and varidase, and (2) number of T-helper/inducer-, T-suppressor/cytotoxic-, total T-, and natural killer cells in the peripheral blood. After a course of immunotherapy no changes in the mean value of the percentage of the cell-subpopulations could be observed, but in a quarter of all patients a reaction in the LMI-test could be shown. In the control group, under chemotherapy, the percentage of T-helper-, T-suppressor- and total T-cells decreased strongly, whereas in the immunotherapy group the percentage of these populations remained unchanged. Under the restriction of a mean observation time of only 18 (5-32) months no differences concerning the survival time between both groups could be observed; only in advanced ovarian cancer patients could a slight positive effect of immunotherapy be shown. These results support the indication of a course of immunotherapy with thymopentin in combination with chemotherapy, especially in advanced cancer patients.

In spite of the efforts made in in-vitro fertilization and embryo transfer in recent years, little progress has so far been made with regard to the pregnancy rate. While embryo transfer rates of over 80% are now being attained, the pregnancy rate achieved with extracorporal fertilization in the majority of study groups is well under 20%. Between 1981 and May 1986, 1088 laparoscopies were performed in our study group with the aim of follicle puncture and subsequent in-vitro fertilization and embryo transfer. A total of 171 pregnancies resulted from 832 embryo transfers. The pregnancy rate has been increased since 1981 to 21.2% at present (1986). Since April 1986 we have been using a modified embryo transfer procedure: in 25 patients embryo transfer has been accomplished under barbiturate anesthesia. Twelve of these patients have become pregnant, i.e., 48% of the embryo transfers. There are signs that this embryo transfer method will lead to a distinct improvement in the pregnancy rate. An attempt should be made to confirm these provisional results in a larger collective.

Considering that in gynecologic carcinomas cells and cellular tumor parts can be left in the abdominal space leading to an increased growth of recurrences due to a change in the cell-kinetic situation, 19 patients with advanced gynecologic tumors were treated locally with 15 mg mitomycin or 30 mg (40 mg) mitoxantrone dissolved in 1000 mg saline before the operation was completed. The course of disease after intraoperative, intraperitoneal chemotherapy given in addition to the usual chemo- or radiotherapeutic regimen was as follows: in 14 patients assessed up to now, a NED state was achieved 4 times, while there was a partial remission in 2 cases and a SD situation in one. In two women there was progression. In the group with recurrent treatment (5 women), a further progressive course was observed in 4 patients. In one case there was a partial remission. With mitoxantrone serum levels between in 10-60 ng/ml, two subilium situations and postoperative nausea were noticed as side-effects. Among the changes noted in the chemical laboratory, there was one leukopenia grade IV and five grade III-leukopenias as well as various intermittent alterations of the liver values and electrocytes. One case of wound healing disturbances with prolapsed small intestine convolution in the wound area was observed in the tumor growth developing in the abdominal wall. The side-effects of the intraoperative and intraperitoneal therapy with cytostatics were generally acceptable. Given the observed courses of the disease, a continuation of the intraperitoneal therapy form aimed against postoperative residual and aggressively growing tumor cells would appear to be justifiable.

Phosphate-dependent changes of the mycelial lipid composition were studied in the streptothricin-producing parental strain Streptomyces noursei JA 3890 b/2 and its mutant RG 2. In contrast to its ancestor, the mutant was capable of producing the antibiotic nourseothricin even when large quantities of inorganic phosphate were present in the medium. The apparent insensitivity of the secondary metabolism to phosphate inhibition corresponds to a decreased level of phospholipids in the presence of excessive inorganic phosphate and, during phosphate limitation, to a much higher production of the alkaline phosphatases. A model is discussed which proposed the control by a common genetic element of both the phospholipid and antibiotic production.

Zero time addition of the autoregulator (L-factor) from S. griseus (Lkm+Amy+) to surface cultures of its bald mutant ZIMET 43 682 (Amy-Lkm-) restored the capacity to form both anthracycline-type antibiotic leukaemomycin and aerial mycelium. The pertinent mycelia displayed the same growth rate and cellular levels of nucleic acids as the asporogeneous phenotype but the composition of fatty acids and phospholipids as well as the ratio of cytochromes b and c were altered. These differences indicate alterations in the cellular architecture of substrate and aerial hyphae. The results suggest that the autoregulator triggers the onset of a complex programme of differentiation at a very early stage.

Since it became clear that tumor cell colonies growing in soft agar are closely related to embryonic tumor cells, it has been possible to examine, with the aid of immunohistochemical methods, the behavior of estrogen receptors (ER) and lectin binding sites on those tumor components which determine malignant growth. With this aim in mind, 14 breast cancers, four ovarian cancers, and one corpus cancer with ER concentrations known from DCC studies were cultured in a colony assay. Using immunohistochemical techniques it was possible, after preparing permanent cultures, to demonstrate the estrogen receptors and lectin binding sites on the tumor cell colonies cultured in vitro.

Mutants of Streptomyces griseus blocked in cytodifferentiation regained their capacity to form differentiated mycelia and/or anthracycline pigments in the presence of butyrolactone-type autoregulatory effectors such as trans-2-(6'-methylheptanol-1'-yl)-3-hydroxymethyl-4-butanolide+ ++. In the pertinent indicator strains, the effect has been correlated with the increase of lipid synthesis, with changes in the composition of lipid fraction and with the restoration of the production of neutral proteinases. The results suggest that autoregulatory butyrolactones from streptomycetes stimulate cytodifferentiation of their producers at an early stage of development.

In normal pregnancies the determination of the L/S ratio permits a reliable ante-partum evaluation of fetal maturity. Good correlation between ante-partum determination and the condition of the newborn is obtained. In the 566 L/S ratios obtained from abnormal pregnancies the method was also reliable. There was 217 Rh incompatabilities, 51 pre-eclampsias, 31 diabetic patients, 112 premature labours, 78 placental insufficiencies and 77 premature ruptures of the membranes. In patients with diabetes the ante-partum diagnosis of pulmonary maturity is doubtful. In some patients with diabetes pulmonary maturity appears to be delayed. A difference between the abnormal pregnancies was not recognized. In the different types of high risk pregnancies no acceleration or retardation of pulmonary maturity was detected with statistical significance. However, in some cases of premature rupture of the membranes, premature labour and placental insufficiency, pulmonary maturity appears to be accelerated.

The advantages of caudal anaesthesia in obstetrics far outweigh the few disadvantages. The perinatal mortality of premature infants is reduced under caudal anaesthesia. Breech deliveries are easier under caudal anaesthesia. In these cases regional anaesthesia is considered an obstetrical indication. It is possible to carry out all vaginal and abdominal procedures with this anaesthesia. The elimination of pain during labor and delivery permits a more intensive experience of labor and delivery for the mother under caudal anaesthesia.

In-vitro fertilisation of human egg cells and subsequent embryonal transfer offers a possibility to sterile women with oviducts incapable of functioning, to become pregnant after all. This article describes the performance of laparoscopic follicular puncture and extraction of egg cells for in-vitro fertilisation. Laparoscopic follicle puncture as described here was performed from July 1982 to March 1983 on 130 patients at the Gynaecological and Obstetric Hospital in Lübeck. 585 follicles were punctured and 404 egg cells obtained, 210 of which met the criteria of a pre-ovulatory egg cell. Of the 210 pre-ovulatory egg cells, 177 could be fertilised. In 95 patients, embryo transfer was effected in the 2-cell to 8-cell stage. The fertilisation rate obtained was 84%, with an embryo transfer rate of 71%. Since this patient group also included andrological causes of sterility, this figure can be considered quite satisfactory, which is decisively due, among other things, to the puncture technique described here. Pregnancy was attained in 14 cases so far. It is certain that the pregnancy rate can be improved still further.

The occurrence of a premature luteinizing hormone (LH)-surge during gonadotropin stimulation for in-vitro fertilization leads to cancellation of the cycle. Moreover, insufficient follicular maturation is often caused by elevated basal gonadotropin levels. Therefore the gonadotropin releasing hormone (GnRH) agonist, D-TRP6-LHRH, was applied to patients exhibiting premature LH-surges, hyperandrogenemia or incipient premature menopause. 119 cycles were treated, using a long-acting versus a short-acting GnRH agonistic analogue. In protocol 1, patients received daily subcutaneous injections of 100-500 micrograms of a short-acting compound. In protocol 2, a long-acting bolus of 3.2 mg was given intramuscularly. Concomitant human gonadotropin (HMG) stimulation started in protocol 1 after clinical and biochemical evidence of pituitary suppression and in protocol 2 after a fixed suppression interval of 14 days. In protocol 1 higher estrogen levels were reached with more oocytes harvested. The pregnancy rate per transfer was increased from 3.5% to 18%, with most pregnancies occurring with protocol 2. The cancellation rate of 13.4% was mainly due to insufficient follicular development in patients, in whom premature menopause was suspected. Hyper-androgenemic patients with an elevated LH/FSH-ratio exhibited the best follicular recruitment with the highest pregnancy rate of 25% per transfer. Thus combined GnRH-agonist/gonadotropin stimulation offers a causal treatment for patients susceptible to premature LH-surges and for hyperandrogenemic patients.

The psychophysiological responses of 60 subjects were measured as they observed a performer play a roulette game. Half of the subjects were led to believe that they were similar to the performer in personality and values, and half were led to believe that they were dissimilar. Half of the subjects in each condition believed that the performer won money and experienced pain as he played the game, and half believed that he performed a cognitive and motor skill task. Subjects who observed a performer who ostensibly experienced pleasure and pain exhibited greater psychophysiological reactions than subjects who did not. Subjects who believed they were similar to the performer tended to react more strongly than subjects who believed they were different from him. Similar subjects also reported identifying most with the performer and feeling the worst while he waited to receive shocks. It was concluded that the similar subjects empathized most with the performer who appeared to experience pleasure and pain. When required to make a choice between helping themselves at a cost to the performer or helping the performer at a cost to themselves, the subjects who reacted most empathically behaved most altruistically. The results were interpreted as casting some light on century-old questions about the human capacity for altruism.

Microlymphocyte transformation tests were done on 33 female rabbits immunized with different components of rabbit ejaculate. In rabbits injected with total ejaculate or washed spermatozoa a specific stimulation of lymphocytes could be demonstrated by exposure to washed spermatozoa. It is shown that it is necessary to notice the length of time for cultivation, the antigen concentration and especially the time-interval after the last immunization.

A new method for determining fetal lung maturity is discussed. Molecular mobility can be determined spectrophotometrically by measuring the fluorescent polarisation (FP) in lipid solutions. This method of determining the microviscosity in lipid solutions can now be applied also in the amniotic fluid. Measurement of FP by means of an electro-optical microviscosimeter is proportional to the lipid concentration in the amniotic fluid and is in good correlation with fetal lung maturity. FP was measured between the 17th and 41st weeks of gestation in 147 amniotic fluid samples. During pregnancy the FP value drops significantly from 0.350 to 0.200. A FP value of 0.290 was fixed as the limit value beyond which RDS may be expected to occur. With a limit value of 0.290, the RDS identification probability was calculated at 94%, whereas the calculated probability of RDS was 81% and the exclusion probability of RDS 97%. At the same time, the probabilities, calculated on the parallel determination of the L/S ratio, yielded a slightly more unfavourable result. The drop in the FP level correlates significantly with the increase of the L/S ratio in the investigated amniotic fluid samples (correlation coefficient 0.77). The method, which requires very special equipment, offers many advantages: it is very accurate, simple, well reproducible and can be performed within half an hour at relatively low laboratory cost.

We have studied the presence of polychlorinated biphenyls in human body fluids associated with reproduction. Since the polychlorinated biphenyls represent a family of compounds, 3 of the main congeners of this family were selected for this study. The distribution of these 3 congeners was investigated in 37 specimens of follicular fluid and in 16 specimens of sperm fluid. Both fluids showed a similar, low contamination with total polychlorinated biphenyls (ca. 10 micrograms/kg on average), but it was evident that the follicular fluids preferentially accumulated the more highly chlorinated components. This finding must be taken into account when interpreting the concentration levels of the main congeners in relation to total pollution and the toxic potential of polychlorinated biphenyls.

Assuming that cells and portions of tumor may remain in the abdominal cavity after surgery to reduce tumor size in cases of ovarian carcinoma, and that a change in cell kinetics could result in accelerated growth in the event of a recurrence, 23 patients with advanced tumors were given local (intraperitoneal) treatment intraoperatively. The treatment consisted of 15 mg Mitomycin C or 30 or 40 mg of Mitoxantron, in 1000 ml normal saline. Since the observation time was so short, the tolerance and side effects of this form of treatment were of primary interest, rather than remission quotas and survival times. The principal abdominal complaints included two subileus conditions which responded well to therapy and the problem of postoperative nausea. Four patients reacted to the treatment described with temperatures of over 38 degrees C. Chemical changes detected in the laboratory included 18 cases of leukopenia, which in one case reached WHO Grade 4. Intermittent changes in liver values and electrolytes were observed in isolated cases. Wound-healing impairments occurred in three cases. In one of them, a patient who sustained a prolapse of the small intestine with tumor growth into the abdominal wall, reoperation was necessary. Taken overall, the side effects of the intraoperative, intraperitoneal cytostatic therapy were acceptable. In view of the courses observed and with the idea of employing a form of therapy to combat aggressive growth of tumor cells remaining after surgery, it appears justified to continue with this form of treatment.