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Background: For a number of decades, hairy cell leukemia (HCL) has been linked with polyarthritis, vasculitis, symptomatic cytopenias and thrombosis in the medical literature. Notwithstanding, the significance of these associations has not been well understood. Therefore, we have decided to analyze them further., Methods: We provide herein a comprehensive literature review of the prevalence of autoimmune disorders in patients with HCL. Most relevant publications were identified through searching the PubMed/Medline database for articles published from inception to February 2014., Findings: Perhaps due to the rarity of HCL, scientific literature on autoimmune conditions in patients with HCL consists mainly of published case series and isolated reports. Our analysis identified increased prevalence of various autoimmune conditions in patients with HCL, including various vasculitides, immune cytopenias and antiphospholipid antibody syndrome (APS) among others., Conclusions: Presence of certain autoimmune disorders should increase the suspicion of HCL in an appropriate clinico-laboratory context. Conversely, the diagnosis of HCL should prompt early recognition of certain autoimmune disorders if clinical suspicion exists. While some of these autoimmune diseases are thought to be secondary to the dysfunctional immune response associated with underlying malignant process, others could be primary and might even play a role in the HCL pathogenesis. The autoimmune complications can pose important clinical problems for the HCL patients. Therefore, a catalogue of these problems is important for alerting physicians to watch for them and diagnose them promptly.

Background: Increased risk of B-cell lymphoma in patients with autoimmune diseases is well known. Medical literature also links various autoimmune conditions with T-cell lymphomas (TCLs), a rarer but very heterogeneous group of generally aggressive neoplasms., Methods: Herein, we provide a comprehensive analysis of the available peer-reviewed literature on epidemiology, clinico-laboratory features and management of autoimmune diseases in patients with TCL. Most relevant publications were identified through searching the PubMed/Medline database for articles published from inception to June 2014., Findings: Prevalence of various hematologic and non-hematologic autoimmune conditions in patients with TCL appears to be increased. Due to the rarity of TCL, scientific literature on autoimmune conditions in these patients consists mainly of case series and isolated reports., Conclusions: Autoimmune disorders can develop prior to, during or after the onset of TCL. The diagnosis of TCL should prompt early recognition of certain autoimmune disorders if clinical suspicion exists. A dysfunctional immune response in these patients may be responsible for the occurrence of autoimmunity. Conversely, autoimmune conditions might create a favorable milieu for T-cell lymphoma pathogenesis. Therefore, their presence should increase the suspicion of TCL in an appropriate clinico-laboratory context. There is need for larger studies to assess further the TCL-autoimmunity relationship as prognosis and management of these patients can be considerably affected.

Our purpose was to determine the dynamics of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) carriage and its determinants in persons working at pig farms, in order to identify targets for interventions. This prospective cohort study surveyed 49 pig farms in the Netherlands on six sampling dates in 1 year (2010-11). Nasal and oropharyngeal swabs were collected, as well as environmental surface samples from stables and house. Of 110 pig farmers, 38% were persistent MRSA nasal carriers. The average cross-sectional MRSA prevalence was 63%. Methicillin-susceptible S. aureus (MSSA) nasal carriage was associated with fewer MRSA acquisitions (prevalence rate (PR) = 0.47, p 0.02). In multivariate analysis, an age of 40-49 years (PR = 2.13, p 0.01), a working week of ≥40 h (PR=1.89, p 0.01), giving birth assistance to sows (PR=2.26, p 0.03), removing manure of finisher pigs (PR=0.48, p 0.02), and wearing a facemask (PR = 0.13, p 0.02) were significantly related with persistent MRSA nasal carriage. A higher MRSA exposure in stables was associated with MRSA in pig farmers (p <0.0001). This study describes a very high prevalence of LA-MRSA carriage in pig farmers, reflecting extensive exposure during work. We identified the possible protective effects of MSSA carriage and of continuously wearing a facemask during work., (© 2014 The Authors Clinical Microbiology and Infection © 2014 European Society of Clinical Microbiology and Infectious Diseases.)

There are indications that livestock-associated MRSA CC398 has a reduced human-to-human transmissibility, limiting its impact on public health and justifying modified control measures. This study determined the transmissibility of MRSA CC398 from livestock veterinarians to their household members in the community as compared to MRSA non-CC398 strains. A one-year prospective cohort study was performed to determine the presence of MRSA CC398 in four-monthly nasal and oropharyngeal samples of livestock veterinarians (n  =  137) and their household members (n  =  389). In addition, a cross-sectional survey was performed to detect the presence of MRSA non-CC398 in hospital derived control patients (n  =  20) and their household members (n  =  41). Staphylococcus aureus isolates were genotyped by staphylococcal protein A (spa) typing and multiple-locus variable-number tandem repeat analysis (MLVA). Mean MRSA CC398 prevalence over the study period was 44% (range 41.6-46.0%) in veterinarians and 4.0% (range 2.8-4.7%) in their household members. The MRSA CC398 prevalence in household members of veterinarians was significantly lower than the MRSA non-CC398 prevalence in household members of control patients (PRR 6.0; 95% CI 2.4-15.5), indicating the reduced transmissibility of MRSA CC398. The impact of MRSA CC398 appears to be low at the moment. However, careful monitoring of the human-to-human transmissibility of MRSA CC398 remains important.

Kikuchi-Fujimoto disease (KFD), also known as histiocytic necrotizing lymphadenitis, is a rare, self-limited disease of unknown etiology. This condition is most commonly encountered in Asian and Caucasian females of childbearing age who usually present with cervical lymphadenopathy and fever. Though rarely, KFD has been described in younger African-American females as well. It often mimics more serious conditions such as lymphoma, metastatic solid malignancy, HIV infection, tuberculosis, sarcoidosis, or systemic lupus erythematosus. Although its etiopathogenesis has not been fully elucidated, literature suggests viral or possibly autoimmune components to play a role. We describe a 34-year-old African-American female who presented with constitutional symptoms and polyadenopathy on clinical examination and imaging, of which the portacaval and portahepatis lymph nodes were most prominent. An extensive workup was otherwise unremarkable, and biopsy showed histiocytic necrotizing lymphadenitis. Initially, her clinical condition improved spontaneously, and she required only a short course of oral steroids. Three months later, she relapsed with bilateral cervical adenopathy and constitutional symptoms and was successfully managed again with steroids. Our case is unique with respect to (a) portahepatis and portacaval node enlargement as the dominant adenopathy and (b) her underlying conditions of fibromyalgia and chronic fatigue syndrome.

Methicillin-resistant Staphylococcus aureus (MRSA) has rapidly emerged worldwide, affecting both healthcare and community settings, and intensive livestock industry. The efficient control of MRSA strongly depends on its adequate laboratory detection. This guideline provides recommendations on the appropriate use of currently available diagnostic laboratory methods for the timely and accurate detection of MRSA in patients and healthcare workers. Herewith, it aims to standardise and improve the diagnostic laboratory procedures that are used for the detection of MRSA in Dutch medical microbiology laboratories.

We screened a cohort of 5 male and 20 female patients with a Rett spectrum disorder for mutations in the coding region of FOXG1, previously shown to cause the congenital variant of Rett syndrome. Two de novo mutations were identified. The first was a novel missense mutation, p.Ala193Thr (c.577G>A), in a male patient with congenital Rett syndrome, and the second was the p.Glu154GlyfsX301 (c.460dupG) truncating mutation in a female with classical Rett syndrome, a mutation that was previously reported in an independent patient. The overall rate of FOXG1 mutations in our cohort is 8%. Our findings stress the importance of FOXG1 analysis in male patients with Rett syndrome and in female patients when mutations in the MECP2 and CDKL5 genes have been excluded.

This paper describes a methodology based on hollow-cone dark-field (HCDF) transmission electron microscopy (TEM) to study dislocation structures in both nano- and micro-crystalline grains. Although the conventional approach based on a two-beam condition has been commonly used to acquire weak-beam dark-field (WBDF) TEM images for dislocation structure characterization, it is very challenging to employ this technique to study nanocrystalline materials, especially when the grains are less than 100 nm in diameter. Compared to the conventional two-beam approach, the method described in this paper is more conducive for obtaining high-quality WBDF-TEM images. Furthermore, the method is suitable for studying samples with both nanocrystalline and coarse-grains. A trimodal Al metal-matrix-composite (MMC) consisting of B(4)C particles, a nanocrystalline Al (NC-Al) phase, and a coarse-grained Al (CG-Al) phase has been reported to exhibit an extremely high strength and tailorable ductility. The dislocations in both NC-Al and CG-Al phases of the trimodal Al MMCs at different fabrication stages were examined using the HCDF method described. The influence of the dislocation density in both NC-Al and CG-Al phases on the strength and ductility of the composite is also discussed., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

In 2001, analysis of Swiss data collected since 1993 included 1001 treatment cycles with IVF, 2217 treatment cycles with intracytoplasmic sperm injection and 2160 treatment cycles with frozen-thawed embryos or zygotes. IVF cycle number has remained constant over the past 10 years, now representing only 18% of the total. ICSI treatment cycles have plateaued since 2001. Altogether, patients receive 1.56 treatment cycles per year, nearly constant since 1995. Mean maternal age has increased from 33.9 to 35.7 years, while mean number of recovered oocytes has increased by 1.3. Considerable improvement was seen in clinical pregnancy rate after 'fresh' treatment cycles since 2000. Mean number of replaced embryos in 'fresh' treatment cycles has fallen to below 2.5 since 1996, long before the legal imposition of the three-embryo transfer limit in 2001, and is still decreasing without affecting the consistent twin pregnancy rate of 19%. The frequency of ovarian hyperstimulation syndrome has increased three-fold. External audits have reduced the mean number of errors per data file by half, and increased the number of correct files by 20%. Data collected over this 10-year period show that despite the introduction of a restrictive law and increasing mean maternal age, the overall clinical pregnancy rate has continued to improve.

Chromosomal mosaicism has been reported in in vitro-cultured embryos at early cleavage stages, as well as in morulae and blastocysts. We have assessed the incidence and pattern of mosaicism during in vitro development of human embryos from early-cleavage stages to morula and blastocyst. Fifty spare embryos were fixed for fluorescence in situ hybridization (FISH) analysis for chromosomes X, Y, 13, 18, and 21 on days 2 or 3 (4- to 10-cell stage) (n = 16), on day 4 (morula stage) (n = 14), on day 5 (pre-expanded blastocyst) (n = 5), and the expanded blastocyst stages (n = 15). Blocked embryos (no cleavage observed within the last 24 hr) were not included. A total of 2367 cells were analyzed. Four early-cleavage stage embryos were found uniformly diploid; all of the others were mosaic for the chromosomes analyzed (mean diploid nuclei 48.3% +/- 28.7). All of the embryos at more advanced developmental stages, except one fully normal morula, had mosaic chromosome constitutions, with an increase in the percentage of diploid cells in morulae, pre-expanded, and expanded blastocysts, respectively (mean diploid nuclei 78.6% +/- 11.7, 66.0% +/- 20.8, 79.6% +/- 12.8), in comparison with earlier stages. Hypotheses about the origin of mosaicism and embryo regulation mechanisms will be discussed.

Purpose: To compare the frequency and the degree of mosaicism in human embryos from Robertsonian translocation (RT) t(13;14) carriers, with embryos from karyotypically normal IVF patients, Methods: FISH analysis of embryos from PGD cycles for RT t(13;14), with probes for chromosomes 13, 14, and 18 (Group I) and of embryos from karyotypically normal IVF patients with probes for chromosomes 13, 18, 21, X, and Y (Group II)., Results: The incidence of abnormal mosaic embryos was significantly higher in group I (38/51) as compared with group II (6/45) (chi2: P < 0.01). Furthermore, in group I the percentage of diploid cells per embryo was lower for chromosome 13 and 14 in comparison with 18, while in group II no differences were observed between the five chromosomes analyzed., Conclusions: RT induces a high frequency of mosaicism specifically for the chromosomes implicated in the translocation; the analysis by FISH of two blastomeres is strongly recommended for these patients

Background: The use of mechanical and enzymatic techniques to isolate preantral follicles before in-vitro culture has been previously described. The aim of this study was to assess the effect of the isolation procedure of mouse preantral follicles on their subsequent development in vitro., Methods: Follicles were isolated either mechanically or enzymatically and cultured using an individual non-spherical culture system. Follicular development and steroidogenesis, oocyte in-vitro maturation and embryo development were assessed for both groups., Results: After 12 days of culture, follicles isolated mechanically had a higher survival rate but a lower antral-like cavity formation rate than follicles isolated enzymatically. Enzymatic follicle isolation was associated with a higher production of testosterone and estradiol compared with mechanical isolation. A stronger phosphatase alkaline reaction was observed after enzymatic isolation, suggesting that follicles isolated enzymatically had more theca cells than those isolated mechanically. However, both isolation techniques resulted in similar oocyte maturation and embryo development rates., Conclusions: Enzymatic follicular isolation did not affect theca cell development. Follicular steroidogenesis was enhanced after enzymatic isolation but the developmental capacity of oocytes was comparable to that obtained after mechanical isolation.

Purpose: To simplify clinical routine and to improve medical quality without exceeding the existing resources. Intensifying communication and cooperation between all institutions of patients' health care. The huge amount of documentation work of physicians can no longer be done without modern tools of paperless data processing., Methods: The development of ODS was a tight cooperation between physician and technician which resulted in a mutual understanding and led to a high level of user convenience. - At present all cases of gynecology, especially gynecologic oncology can be documented and processed by ODS. Users easily will adopt the system as data entry within different program areas follows the same rules. In addition users can choose between an individual input of data and assistants guiding them through highly specific areas of documentation., Results: ODS is a modern, modular structured and very fast multiuser database environment for in- and outpatient documentation. It automatically generates a lot of reports for clinical day to day business. Statistical routines will help the user reflecting his work and its quality. Documentation of clinical trials according to the GCP guidelines can be done by ODS using the internet or offline datasharing., Conclusions: As ODS is the synthesis of a computer based patient administration system and an oncological documentation database, it represents the basis for the construction of the electronical patient chart as well as the digital documentation of clinical trials. The introduction of this new technology to physicians and nurses has to be done slowly and carefully, in order to increase motivation and to improve the results.

Mosaicism in 4-8-cell human embryos analysed by fluorescence in situ hybridisation (FISH) has been widely reported, but few studies have addressed the incidence of mosaicism in more advanced embryonic stages. In the present study we analysed spare human embryos in a case of preimplantation genetic diagnosis (PGD) for increased risk of aneuploidy because of an infertile 47,XYY man. After replacement of two embryos typed as 1818XX at PGD, six spare embryos (not frozen because of their low quality) were re-analysed on Day 5 for PGD confirmation. Out of five embryos typed as 1818XY at PGD, four were diploid mosaic (DM) and one was normal in all cells. The sixth embryo, typed as 18XYY/1818181818X at PGD, was a DM. In spite of the bias of our small series of morphologically low-quality embryos, the surprisingly high proportion of mosaics (which confirms previous findings) questions the validity of PGD, but supports the strategy of transferring only the embryos where two blastomeres gave normal and concordant results at PGD. More data are required to understand the clinical significance of early diploid mosaicism (and its impact on implantation rate) and to determine whether some diploid mosaic embryos might be considered safe for transfer., (Copyright 2000 John Wiley & Sons, Ltd.)

Objective: To improve in vitro culture conditions and human embryo selection before transfer after IVF with intracytoplasmic sperm injection (ICSI)., Design: A controlled, randomized, prospective study., Setting: University hospital-based IVF-ET program., Patient(s): Couples undergoing ICSI., Intervention(s): Culture of human embryos in the presence of 1 mM or 5.56 mM glucose and metabolic measurements with the use of noninvasive microfluorescence assays immediately after ICSI to the time of transfer., Main Outcome Measure(s): Embryo development, implantation rate, and glucose and pyruvate uptake., Result(s): Fertilization rates, early embryo development, and implantation rates were not significantly different between 1 mM and 5.56 mM glucose. Pyruvate uptake was significantly higher during the formation of the pronuclei, at 15 +/- 0.7 and 11.4 +/- 1.3 pmol/embryo/h for fertilized and unfertilized oocytes, respectively. Pyruvate uptake did not correlate with cleavage stage or embryo morphology. However, during the second day of incubation, pyruvate uptake was significantly higher for the untransferred embryos of pregnant women compared with nonpregnant women, at 17.9 +/- 1.5 and 10.8 +/- 1.0 pmol/embryo/h, respectively., Conclusion(s): The increased level of pyruvate uptake during fertilization reflects the increased demand for energy necessary for the formation of the pronuclei. However, the metabolic measurements could not improve the selection of embryos with the best implantation potential. Finally, the reduction of glucose concentration in the culture medium failed to improve embryo viability.

The introduction of the Intracytoplasmic Single Sperm Injection (ICSI) has been a turning point for the treatment of severe male infertility. ICSI allowed not only to reduce fertilization failure from 35% to 0.7% but created at the same time the opportunity for a group of patients with extremely low sperm counts to procreate. The discovery that breaking the tail of the spermatozoon prior to the injection was the most important step is at the origin of major improvements: fertilization increased from 22% to 77%, pregnancy rate from 16% to 54% and the implantation rate from 7.4% to 26%. From October 1994 to April 1999, 835 ICSI cycles were performed and resulted in 312 ongoing pregnancies (37%), fertilization rate was 75%, with a fertilization failure of only 0.7%. The use of ICSI and IVF on sibling oocytes for semen samples with doubtful fertilizing ability clearly illustrated the superiority of ICSI. No fertilization failures occurred after ICSI and the fertilization rate was 76% versus 27.8% (P < 0.01). Similar benefit of ICSI was shown for crytozoospermia up to then a hopeless situation. A total of 26 pregnancies were obtained out of 87 cycles with a fertilization rate of 58.8%. Similar results were obtained when ICSI was combined with testicular sperm, 20 pregnancies occurred after from 46 transfers (43%) including cycles with cryopreserved testicular sperm. It is now clear that ICSI is the method of choice for the treatment of severe male infertility.

This contribution summarize ten years of in vitro fertilization of clinical work. Activity growth, improvements of results (mean fertilization rate increased from 45% to 58%, fertilization failure dropped from 18% to 7%, pregnancy chances gains 9% to reach 44% per trial) and new treatments possibilities (severe male infertility) thanks to the ICSI technic were the major characteristics of this last ten years. The original anonymous oocyte donation program with donors permutation initiated as soon as 1990 has imposed itself due to it's exceptional efficiency with a pregnancy rate of 95% per oocyte pick up on a population of 46 donors and 145 recipient cycles. Thanks to the large population studied (4028 cycles, 1071 pregnancies), the tendencies in human fecundity (impact of age) and the risks linked to multiples pregnancies could be highlighted, stressing the importance of future developments presented in the other contributions following this general presentation of results.

Objective: Our objective was to analyze the outcome of cryopreserved embryos obtained after intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF) in terms of survival rate, implantation rate (IR), total and clinical pregnancy rate (PR) in a retrospective, comparative study., Methods: Three hundred seventy-five IVF and 463 ICSI surnumerary cleaved embryos, frozen on Day 2 with 1,2-propanediol, were thawed., Results: Thirty-two percent of the thawed IVF embryos survived and 11 pregnancies (8 clinical) were obtained from 68 transfers (16.1%). Fourty-seven percent of the ICSI embryos survived, with 19 pregnancies (18 clinical) from 116 transfers (16.4%). The IR was 8.5% (8/94) in IVF cycles and 10.8% (20/185) in ICSI cycles., Conclusions: A significantly better survival rate of ICSI embryos was observed but with no difference in PR, preclinical, and clinical abortion rate, or IR.

Purpose: Our purpose was to compare the embryo culture performance of two types of petri dishes (Nunc and Falcon)., Methods: Mouse zygotes were cultured up to the expanded blastocyst stage in both types of dishes. The oocytes from 50 in vitro fertilization cycles were randomly divided between the two types of dishes. Fertilization, cleavage, and embryo quality were compared. Oocytes from another 50 cycles were all cultured at random in either type of dish. Pregnancy and implantation rates were compared between the two types., Results: Of 91 mouse zygotes, 81 cleaved to two-cell-stage embryos, and 64 became expanded blastocysts in Falcon dishes; of 99 zygotes, 81 cleaved to two-cell-stage embryos and 66 became expanded blastocysts in Nunc dishes. Of 248 oocyte-cumulus complexes (OCC), 145 fertilized in Falcon dishes, and of 269 OCC, 175 fertilized in Nunc dishes. The high quality embryo ratio was 51 out of 118 in Falcon dishes, not different from that in Nunc dishes, 58 out of 139. In Falcon dishes 72 out of 118 embryos were at least at the four-cell stage after 45 hr, versus 70 out of 139 in Nunc dishes. Twenty-three clinical pregnancies were obtained in the first 50 cycles with sibling oocytes. In the second group, with randomization of the cycles between Nunc and Falcon, 8 pregnancies were obtained in the Nunc and 10 in the Falcon dishes. The implantation rate in this second group of 50 cycles was 9 out of 61 in Falcon and 11 out of 57 in Nunc dishes.

A case of invasive pulmonary aspergillosis in an allogeneic bone marrow transplant recipient caused by Aspergillus ustus is presented. A. ustus was also recovered from the hospital environment, which may indicate that the infection was nosocomially acquired. A literature review revealed seven cases of invasive infections caused by A. ustus, and three of these were primarily cutaneous infections. In vitro susceptibility testing of 12 A. ustus isolates showed that amphotericin B and terbinafine had fungicidal activity and that itraconazole and voriconazole had fungistatic activity.

Objective: We analyzed retrospectively the relevance of 4 years of quality control on homemade culture medium with the mouse IVF and zygote bioassay., Design: In vitro or in vivo fertilized mouse oocytes were cultured in each batch of medium. Two-cell-stage and expanded blastocyst development was recorded for each batch of medium. Data on fertilization and embryo quality obtained in human in vitro fertilization were recorded for each batch. IVF treatment cycles for male infertility and cycles with sperm microinjection were excluded., Results: Human oocyte fertilization dropped from 60 to 54%, respectively, from 57 to 41% in a significant way (P < 0.05 resp. P < 0.01) and the human mean embryo score decreased from 4.17 +/- 1.21 to 3.69 +/- 1.06 (P < 0.05) when media were used with a low two-cell-stage development (< or = 75%) for the mouse zygote or mouse IVF bioassay. The pregnancy rate was not affected., Conclusions: Media with high scores in mouse bioassays show higher fertilization rates and better embryo quality when used for human IVF.

Objectives: To assess which clinical parameters influence human oocyte zona pellucida (ZP) thickness., Patients: Sixty-five couples undergoing 75 IVF-ET cycles., Main Outcome Measure: Zona pellucida thickness of 827 oocytes measured 16 to 20 hours after in vitro insemination under inverted light microscope., Results: Zona pellucida thickness was 18.9 +/- 3.8 microns (mean +/- SD) for unfertilized, 16.4 +/- 3.1 microns for fertilized, and 15.1 +/- 2.4 microns for polyspermic oocytes (significantly different). Among our patients, a few underwent two (or even three) IVF-ET cycles, and the mean ZP thickness was, in most cases, not significantly different from one cycle to the other(s). Regression analyses were calculated between ZP thickness and available clinical parameters, i.e., the age of the women, the duration of stimulation, the cumulus maturity, the number of retrieved oocytes, the number of hMG doses, the maximum E2 level, and the follicular volume. A significant linear decreasing relationship exists between the mean ZP thickness of each patient and the maximum E2 level and an increasing one with the hMG dose. Relationships with the other parameters appeared to be nonsignificant., Conclusion: The ZP thickness is basically an individual feature that influences the fertilization rate. Nevertheless, it may be influenced slightly by the hormonal treatment during stimulation.

Objectives: To determine the efficacy and innocuousness of long-acting versus short-acting GnRH analogues (GnRH-a) in long protocol for in IVF-ET., Design: Prospective randomized study., Setting: The IVF unit at an academic hospital., Patients: One hundred couples admitted for their first IVF-ET attempt., Main Outcome Measures: Serum concentrations of LH, E2, and P during the all cycles and duration of pituitary desensitization were assessed, as well as fertilization rate, embryo quality, and implantation and pregnancy rates., Results: Significantly more days (10.8 +/- 1.8 versus 9.2 +/- 1.7 days) of stimulation and more ampules of hMG (47 +/- 22 versus 33 +/- 16) were necessary to obtain similar numbers of embryos of quality with the long-acting GnRH-a. Implantation and delivery rates were significantly lower with the long-acting GnRH-a (32.8% versus 21.1%; 48.9% versus 29.1%, respectively)., Conclusions: As the long-acting GnRH-a might interfere with the luteal phase and embryo development, short-acting GnRH-a should be preferred for ovarian hyperstimulation in IVF-ET.

Before starting with the clinical application of ICS, aged unfertilized oocytes were gathered for training and were injected with a single sperm or without a spermatozoon as a control group for activation. Oocyte damage, initially as high as 40% was reduced to 15% after 60 oocytes. Normal fertilization (2PN) occurred in 18% of the injected oocytes. After this training period 1,488 metaphase II oocytes collected during 144 cycles were used for ICSI. Results were split up in 3 periods (n = 55, n = 24, n = 57) corresponding to the different improvements made in the technique. Results form ICSI in combination with MESA (n = 6) were analysed separately. Mean fertilization increased from 24% to 77%. Fertilization failures (18% of the cycles during the first period) vanished in the last period. Implantation rate improved from 7.4% to 11.4% and reached finally 26%. Pregnancy rate per oocyte retrieval was 16%, 25% and 54%. For the MESA group fertilization was 28%, implantation rate 17% and pregnancy rate 33% and only one fertilization failure was observed. A total of 50 pregnancies were obtained including 2 obtained after MESA and 2 with cryopreserved embryos. Four healthy children are born, 9 were early abortions, 37 pregnancies are still on-going. Preclinical practice on aged unfertilized oocytes seems useful before starting with clinical ICSI, as high initial oocyte damage could be reduced and subsequent clinical treatment successfully applied. Offering high fertilization and pregnancy rates in cases of infertility with severe male factor it is extremely worthwhile mastering this new technique.

Objective: Our objective was to evaluate the time course and the predictive value of the extrusion of the second polar body after intracytoplasmic injection (ICSI) related to the fertilization rate, embryo cleavage and quality., Setting: The setting was the in vitro fertilization program of a university hospital., Patients: Twenty-one patients were treated with intracytoplasmic single sperm injection either for fertilization failure in IVF, low fertilization in IVF (< 5%), or severe male factors., Design: One hundred thirty-five of 205 metaphase 2 oocytes treated with intracytoplasmic single sperm injection were observed 1, 2, and 3 hr after the assisted fertilization procedure. Extrusion of the second polar body was recorded. For each of these oocytes, fertilization was noted 18 hr after ICSI and cleavage and embryo quality were assessed 24 hr later. The 70 remaining oocytes were used to assess a possible negative effect of repeated exposure to light microscopy., Results: The extrusion of the second polar body 3 hr after injection was an observation with a sensitivity of 0.87, a specificity of 0.58, and a high positive predictive value (0.90) toward oocyte fertilization. Twenty-nine and four-tenths percent of the oocytes extruded a second polar body within the first hour, 56.6% within the first 2 hr, and 78.3% had a second polar body 3 hr after injection. This time course was related neither to the speed of embryo cleavage nor to the embryo quality. Fertilization, cleavage, and embryo quality were not affected by repeated observation as deduced from comparison with the control group and confirmed by a high pregnancy (62% per oocyte retrieval) and implantation rate (22% per replaced embryo)., Conclusion: Oocytes can be checked, in all safety, 3 hr after a single sperm injection for the presence of a second polar to predict oocyte fertilization with a high certainty.

Objective: Our purpose was to reduce oocyte damage before clinical application of intracytoplasmic single sperm injection by training on aged unfertilized oocytes., Design: Intracytoplasmic single sperm injection (ICSI) was accomplished by micromanipulation of sperm and oocytes., Patients: Thirty-four patients consented to donate unfertilized aged oocytes to train for ICSI. Forty-four patients suffering from severe male infertility were treated with ICSI., Intervention: Oocytes were inseminated by intracytoplasmic single sperm injection., Main Outcome Measures: Oocyte damage and fertilization and pregnancy rates were the outcome measures., Results: One hundred fifty-one aged unfertilized oocytes were gathered for training of which 121 were injected with a single sperm and 30 without a spermatozoon as a control group for activation. Oocyte damage, initially as high as 40%, was reduced to 15% after 60 oocytes. Normal fertilization (2PN) occurred in 18%, and polyploidy in 4.4%. The cleavage rate was 69%; none of these embryos were transferred. In the control group, seven oocytes were damaged, seven (30%) showed one pronucleus, and one showed two pronuclei. No cleavage was observed in the control group. In the clinical trial, 44 patients (61 cycles) were clinically treated with the same ICSI procedure, including 575 of 721 collected oocytes. Damage was 13%, activation was 11%, normal fertilization was 30%, and 5 (1%) polypoid zygotes were observed. The fertilization rate ranged from 5 to 100%, with a mean of 39.5 +/- 4% (SE). Nine patients had no fertilization (15%). Ninety-six percent of the zygotes cleaved and 47% were at the four-cell stage 45 hr after injection. One hundred twelve embryos were replaced in 48 transfers (2.3 embryos/ET). One live birth, one miscarriage, and eight ongoing pregnancies were obtained (22%/ET)., Conclusion: Preclinical practice on aged unfertilized oocytes seems useful before starting clinical ICSI, as the high initial oocyte damage can be reduced and subsequent clinical treatment successfully applied.

Objective: To investigate whether Kremer's sperm-mucus penetration test may predict sperm fertilizing ability in IVF., Design: Kremer's test was prospectively performed on semen samples used for 66 consecutive IVF trials and compared with the fertilization rates and fertilization failure rates observed., Results: Fertilization rates were significantly reduced in cases of abnormal Kremer's test (42% versus 51%; n = 745 oocytes with a statistically insignificant increase in fertilization failure rates (21% versus 10%; n = 66 trials). For abnormal semen, fertilization rates (39% versus 39%; n = 208 oocytes) and fertilization failure rates (20% versus 28%; n = 17 trials) were similar regardless of Kremer's test result. For normal semen, an abnormal Kremer's test implied a significant decrease in fertilization rates (44% versus 54%; n = 537 oocytes) with a statistically insignificant increase in fertilization failure rates (21% versus 6%; n = 49 trials)., Conclusions: Abnormal Kremer's test results identify patients with a decreased in vitro fertilizing ability despite apparently normal semen samples and a group with very low fertilizing failure risk in case of normal semen samples and normal Kremer's test. Kremer's test does not add any predictive value to sperm analysis in the case of abnormal semen samples. These observations point out the importance of the male factor in fertilization failure even in the case of normal semen analysis.

In vitro fertilization and embryo transfer (IVF) may be considered as a particular application of modern medical therapeutics linked to human reproduction. The treatment of human sterility therefore involves some fundamental human values such as life, love and death. The quality of this highly technological treatment with fast knowledge of outcome at the end of the patient's menstrual cycle has been evaluated since the early 80s. It is a typically multidisciplinary team effort involving medical doctors, biologists, laboratory technicians, nurses and clerks that is representative of modern medical practice. IVF covers much more than just embryology, as this review will explain. IVF developed in close relation with clinical and experimental research protocols, which are the major topics of this paper. The newness of the techniques used led to the necessary interactions between clinicians and biologists working on animal experimental embryology.

The clinical results including all in vitro fertilization (IVF) cycles with oocyte pick-up in 1990 are presented. Different types of treatment including classical IVF and embryo transfer, laparoscopic replacement of zygotes in the fallopian tube (ZIFT), IVF with donor sperm (IVF-D), cross fertilization test, embryo freezing, oocyte donation and IVF with epididymal sperm were performed. The total pregnancy rate obtained reaches 38% per oocyte pick-up, 30% of clinical pregnancies (including 4 pregnancies obtained with frozen and thawed embryos). The anticipated "Take Home Baby Rate" will be around 25% per oocyte pick-up, 26 of these 40 pregnancies being today over 20 weeks of gestation. Particular ethical aspects of the program are presented: a study on couple's attitudes regarding embryo freezing as well as the final destination of possibly remaining supernumerary embryos will stress the importance of a precise clear decision on that matter before entering IVF treatment. Indeed the couple's idea on embryo destiny were very precise but also very different. The oocyte donation program has the originality of preserving the donor's anonymity by exchanging the donors recruited by the patients. It will be stressed that this kind of approach combines higher pregnancy chances for the patients, respect of ethical principles linked to gamete donation and gives satisfaction to the patients. The global normalized pregnancy cumulative curve shows that 60% of the couples entering IVF treatment will obtain a child within the first three pick-up cycles.

With the use of a monoclonal anti-glycophorin A antibody and flow cytometric cell sorting, an erythroleukemic bone marrow sample was separated in highly purified erythroblast and myeloblast fractions. Similar karyotypic anomalies were found in both cell populations as in the unseparated bone marrow. This study confirms that acute nonlymphocytic leukemia can originate at the level of a multipotential hemopoietic stem cell.