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Poor-quality medicines are a threat to public health in many low- and middle-income countries, and prospective surveys are needed to inform corrective actions. Therefore, we conducted a cross-sectional survey on a sample of products used for children and available in the private market in Kinshasa, Democratic Republic Congo: amoxicillin (AX) and artemether/lumefantrine (AL), powders for suspension, and paracetamol (PC) tablets 500 mg. Overall, 417 products were covertly purchased from 61 wholesalers. To obtain a representative sample, the products were weighted on their market shares and a subset of 239 samples was randomly extracted to undergo in-depth visual inspection locally, and they were chemically assessed at two accredited laboratories in Belgium. Samples were defined of “poor-quality” if they failed to comply with at least one specification of the International Pharmacopoeia (for AL) or United States Pharmacopoeia 37 (for AX and PC). Results are reported according to the Medicine Quality Assessment Reporting Guideline. The visual inspection detected nonconformities in the aspects of antimalarial powders for suspension, and poor-quality labels across all medicine types. According to chemical analysis, 27.2% samples were of poor quality and 59.5% of AL samples were underdosed in artemether. Poor quality was more frequent for locally manufactured antimalarials (83.3%, P = 0.021; 86.4%, P = 0.022) and PC (4.8%, P = 0.000). The poor quality of the surveyed products may decrease the treatment’s efficacy and favor the development of resistances to antimalarials. It is hoped that these findings may guide the corrective actions of the Democratic Republic of Congo Regulatory Authority, which was the main partner in the research.

The 'ESHRE Guidelines for Good Practice in IVF Laboratories' were drawn up by the Special Interest Group (SIG) in Embryology and published in the year 2000, and since then they constitute the minimal requirements for any laboratory offering assisted reproduction techniques (ART). In the understanding that the embryologist has a responsibility for the correct and justified application of ART in the laboratory, the implementation of these guidelines requires a quality management programme to be in place that encompasses and integrates the operative units, the processes and procedures that represent the core of ART clinics. In March 2004, the European Parliament issued the Directive 2004/23/EC 'On setting standards of quality and safety for the donation, procurement, testing, processing, preservation, storage and distribution of human tissues and cells'. The Directive applies to human tissues and cells, including fresh or frozen reproductive cells for application to the human body, and is mainly concerned with increasing quality and safety through the implementation of a quality management system. Therefore, the European Society of Human Reproduction and Embryology (ESHRE) undertook a series of initiatives aiming to promote assurance of good laboratory practice and to define the concept of qualified embryologists. One ESHRE initiative was to revise the guidelines for good practice in IVF laboratories, not only in response to the need of embryologists for support and guidance in their duties, but also as a complement to the requirements issued by the Tissue and Cell Directive. The SIG in Embryology hopes that this document may assist the laboratory staff to operate according to the requirements of harmonization, implementation, inspection and certification that are now common to all European member states.

Artificial oocyte activation using the calcium ionophore ionomycin is applied successfully in assisted reproduction but some concern exists on the clinical use. The aims of the present study were to optimize the oocyte activation scheme and to address embryo toxicity in a mouse model. Efficiency of oocyte activation and subsequent development was evaluated and ionomycin was found to be an efficient activator at 10 μmol/l. An improved effect of a second exposure to 5 μmol/l ionomycin on blastocyst development was observed. Toxicity of ionomycin on embryos was then investigated by evaluating pre- and post-implantation development of in-vivo fertilized oocytes following exposure to ionomycin. Blastocyst development, blastocyst cell numbers in trophectoderm and inner cell mass were not different between treated and non-treated zygotes. Also implantation rates and fetal parameters such as length, weight and morphological parameters were similar between the fetuses originating from zygotes treated with ionomycin and non-treated zygotes. Furthermore, healthy offspring originating from ionomycin-treated zygotes was born. In conclusion, no adverse effects of ionomycin on in-vitro or in-vivo mouse embryo development were noticed, giving arguments in favour of the use of ionomycin, although negative long-term effects of this compound cannot be excluded at present.

Cryopreservation of ovarian cortical tissue and subsequent transplantation or in vitro culture of follicles are technologies under development with the aim to safeguard fertility in patients with gonadal failure. In the present study, we investigated whether primordial follicles could be triggered to full maturation by a combination of in vivo transplantation and in vitro culture in a mouse model. In a first step, newborn mouse ovaries containing only primordial follicles were allotransplanted under the renal capsule of ovariectomized recipient mice. The second step was to mechanically isolate growing preantral follicles from the graft and culture these in vitro to maturity. In our experiment, one newborn mouse ovary was transplanted under the renal capsule of each 8- to 12-wk-old F1 (C57Bl/6j x CBA/Ca) female ovariectomized recipient (n = 26). Two weeks after transplantation, all 26 grafts were recovered. Four grafts were processed for histology and showed that developmental stages of follicles in 14-day-old ovarian grafts were comparable to those in 14-day-old mouse ovaries. The 22 remaining grafts were used for mechanical isolation of preantral follicles. As a control group, preantral follicles isolated from ovaries of 14-day-old mice were used. The mean preantral follicle yield per ovary was 11 in the transplant group versus 33 in the control group. Follicles were cultured individually in 20-microliter droplets of alpha-MEM supplemented with 100 mIU rFSH and 5% fetal bovine serum for 12 days under an atmosphere of 5% CO(2) in air at 37 degrees C. By Day 12 of culture, 66.5% of follicles retained their oocytes in the grafting group versus 97.5% in the control group (P < 0.001). Final oocyte maturation was induced by addition of 2.5 IU/ml hCG. At 14-16 h post-HCG, the percentages of oocytes showing germinal vesicle breakdown and polar body extrusion were significantly higher in the control group (90.6% and 82.8%) compared to the grafting group (60% and 45%). The mean diameter of the mature oocytes of the grafting group (69.9 +/- 4.45 micrometer) was similar to that of oocytes from the control group (70.5 +/- 2.35 micrometer). Our results suggest that maturation of mouse primordial follicles is feasible by combination of in vivo transplantation and in vitro culture. This two-step strategy may be an attractive model for promoting the growth and maturation of primordial follicles from other species.

This study is a long-term evaluation of the total pregnancy potential of cohorts of fresh and cryopreserved sibling embryos from in-vitro fertilization (IVF) cycles stimulated with either the gonadotrophin-releasing hormone analogue buserelin (BUS) (long protocol) or clomiphene citrate (CC) both in combination with human menopausal gonadotrophin (HMG). Therefore a retrospective analysis was performed on patients who entered the IVF programme between January 1986 and July 1987 and who had triple embryo transfer in the collection cycle. Significantly more fertilized oocytes developed to good-quality embryos in the CC-HMG group (86.1%) than in the BUS-HMG group (80.8%). Transfer of the three morphologically best-looking embryos was performed in day 2 post-insemination in 106 CC-HMG and 80 BUS-HMG cycles. Supernumerary embryos were cultured for a further 24 h and multicellular embryos with up to 20% of fragments were frozen slowly with 1.5 M dimethylsulphoxide on day 3 post-insemination (162 embryos in CC-HMG cycles, 102 embryos in BUS- HMG cycles). Outcome was measured by embryo survival rate, embryo implantation rate and delivery rate in fresh and frozen embryo transfers. Delivery rates were 31.3 and 21.7% per fresh embryo transfer in BUS-HMG and CC- HMG cycles respectively. Fresh embryo implantation rates were significantly higher in collection cycles stimulated with BUS-HMG (17.9%) than in cycles stimulated with CC-HMG (11.3%). Implantation rates were significantly enhanced in embryos transferred in excess of one in cycles leading to pregnancy, perhaps indicative of higher embryo quality in BUS-HMG cycles. Almost all cryopreserved embryos have by now been thawed, so the contribution of frozen embryos to overall pregnancy rates can be evaluated. Overall morphological survival rates of frozen-thawed embryos have by now been thawed, so the contribution of frozen embryos to overall pregnancy rates can be evaluated Overall morphological survival rates of frozen-thawed embryos were similar for 140 embryos from CC-HMG cycles (50%) and 100 embryos from BUS-HMG cycles (46%). The percentage of fully intact embryos was, however, significantly lower in the BUS-HMG group (19%) than in the CC-HMG group (39.5%). Delivery rates were significantly lower following 30 transfers of frozen-thawed embryos from BUS-HMG-stimulated cycles (3.3%) than following 42 transfers of frozen-thawed embryos from CC-HMG cycles (19.1%). Embryo implantation rates were lower for frozen-thawed embryos from BUS-HMG cycles (2.3%) than from CC-HMG cycles (12.7%). Here we demonstrate that ovarian stimulation with the long protocol BUS-HMG instead of the CC-HMG protocol led to higher embryo implantation rates in collection cycles but to lower intact embryo survival rates and to lower embryo implantation rates for frozen sibling embryos. Despite the lower implantation rates with frozen embryos originating from the BUS-HMG protocol, there was no significant difference between total delivery rate per transfer from cycles stimulated with CC-HMG (30.2%) compared with BUS-HMG (33.8%).

OBJECTIVE: To investigate the pregnancy rate of frozen-thawed human supernumerary multicellular embryos that were cryopreserved after intracytoplasmic sperm injection or IVF. DESIGN: A clinical study. SETTING: Consenting patients in an academic research environment. PATIENTS: Couples with severe male infertility, indicated by failed or sporadic fertilization after IVF or subzonal insemination or by

We demonstrated that ovaries removed from female-to-male transsexuals can be used as research material for ovarian cryopreservation, grafting and culture studies. A 21 year old female-to-male transsexual individual, who had been treated with androgens for 12 months, donated her ovaries removed in the sex reversal operation. She had normal numbers of, and proportions of, primordial (98.6%) and primary (1.4%) follicles in her ovarian cortex. After freezing and thawing the ovarian tissue was grafted to immuno-incompetent mice, which were stimulated by human recombinant FSH for 14 days, 10 weeks after transfer. Initiation of growth had occurred in the grafts in which only 79.4% of follicles were primordial, 17.1% primary, and in which there were also secondary and pre-antral follicles. Because long-term androgen treatment does not appear to cause depletion of the primordial follicle pool or affect the developmental capacity of the follicles, ovaries from individuals who undergo sex-reversal operations are an excellent source of tissue for research and maybe even for oocyte donation in the future.

Background Embryo transfer is a crucial step in the IVF process. Most randomized prospective studies comparing transfer catheters have demonstrated significantly higher pregnancy rates with soft versus firm catheters, but none have taken the operator effect into account. Our aim was to perform a prospective randomized clinical trial comparing two catheters and to study interactions between catheters and operators. Methods A prospective randomized trial comparing the Cook K-SOFT-5100 and Frydman classical catheters 4.5 was performed. Three experienced operators participated in the trial, using a fixed distance transfer protocol. Primary end-point was clinical pregnancy rate, secondary end-points were rates of difficult transfer and of catheter failure. Patients were randomized by a computer program prior to embryo transfer. Results A total of 1446 embryo transfers were performed in 1155 women undergoing IVF or ICSI treatment. A total of 723 cycles were randomized to the Cook catheter and 723 cycles to the Frydman catheter. Following intention-to-treat analysis, the adjusted odds ratio of clinical pregnancy between for the Cook versus the Frydman catheter was 1.11 [95% confidence interval (95% CI) 0.89-1.38]. Odds ratios of clinical pregnancy between the Cook and Frydman catheters for the three operators were respectively 1.19 (95% CI 0.84-1.69), 2.35 (95% CI 1.40-3.95) and 0.69 (95% CI 0.48-0.99). Conclusions Variation in pregnancy rates between embryo transfer catheters depends on variation between operators. Results from randomized clinical trials comparing embryo transfer catheters should not be generalized, because inconsistent conclusions may be unavoidable on the account of different proportions of cycles with transfers by each type of operator. The study was registered at clinicaltrials.gov. NCT00766714.

This randomized, controlled trial verifies whether patients with recurrent failed implantation benefit from preimplantation genetic diagnosis for aneuploidy, as compared with conventional assisted reproduction treatment procedures. Two hundred patients with recurrent failed implantation were randomized into two groups. A total of 139 patients underwent ovarian stimulation, and preimplantation genetic screening was performed in 72 patients. Analysis of chromosomes X, Y, 13, 16, 18, 21 and 22 was carried out using fluorescence in-situ hybridization in blastomeres of day-3 cleavage-stage embryos in the study group. The primary endpoint was implantation rate. Secondary endpoints were embryonic morphology and chromosomal status, number of transferred embryos and clinical pregnancy rate. With regard to the implantation rate, there was no significant difference between the study group (21.4%) and the control group (25.3%). The number of embryos transferred was significantly lower in the study group, namely 1.4 (SD 1.0) versus 2.1 (SD 1.0) in the control group (P < 0.05). The clinical pregnancy rate was not significantly different between the groups (25.0% in the study group versus 40.3% in the control group). It can be concluded that preimplantation genetic screening does not increase the implantation rates after IVF-intracytoplasmic sperm injection in women with repeated implantation failure.

OBJECTIVES: To compare the administration of GnRH antagonist in gonadotropin intrauterine insemination (IUI) cycles with cycles where no intervention took place. DESIGN: Meta-analysis of published prospective randomized trials. PATIENTS(S): Five hundred twenty-one patients who were administered a GnRH antagonist and 548 conservatively treated patients who served as control subjects were included in the meta-analysis. INTERVENTION(S): Study selection: Prospective trials were retrieved from Medline and Cochrane Library (last update October 2006). Random effect analysis was used in this meta-analysis. Two independent reviewers performed data extraction. MAIN OUTCOME MEASURE(S): Clinical pregnancy rates. RESULT(S): Six comparisons were retrieved including 1,069 patients. Higher pregnancy rates were found in the randomized controlled trials (odds ratio [OR] 1.56, 95% confidence interval [CI] 1.05-2.33) when a GnRH antagonist was added to a gonadotropin superovulated IUI protocol. Early published studies with smaller sample sizes showed stronger associations (OR 2.31, 95% CI 1.15-4.63) than later studies (OR 1.32, 95% CI 0.79-2.23). CONCLUSION(S): From the randomized controlled trials of this meta-analysis, it is clear that allowing for follicle growth and avoiding premature LH rise, increased pregnancy rates are observed with GnRH antagonist administration. A parallel trend for multiple pregnancy rates in the GnRH antagonist group was observed, although this did not reach statistical significance. The flexible regimen was widely used. This meta-analysis of early data might enhance further research in this direction. Fertil Steril

Objective To submit different glove brands to double-quality control tests using mouse embryo assay (MEA) and the human sperm motility assay (HuSMA). Operator protection against infectious body fluid contamination is a safety issue in assisted reproductive technology (ART). When using gloves in the ART laboratory, toxic substances can be transmitted to culture media, even during brief contact. Design Quality control study of gloves in ART. Setting University hospital-based infertility center. Animal(s) Seven- to 8-week-old female B6D2F1 hybrid mice. Intervention(s) We tested two surgical, two cleanroom, and six examination glove brands. Only gloves brands that passed both HuSMA and MEA were submitted to further QC using zona-free and/or cryopreserved MEA. Main Outcome Measure(s) S perm motility index, two-cell and blastocyst development, blastocyst total cell number. Result(s) Quality control by MEA and HuSMA identified two glove brands to be nontoxic. Conclusion(s) Our study shows that gloves used in ART can be toxic and should be tested as part of an ongoing quality control program.

BACKGROUND: Single-embryo transfer (SET) has proven efficient in reducing multiple pregnancy rates after assisted reproduction technologies (ART). This study compares outcome of singletons after SET and double-embryo transfer (DET). METHODS: We studied 404 SET and 431 DET patients, who delivered a singleton child of >500 g after fresh embryo transfer in a first, second or third cycle. Preterm birth and low birthweight incidences and gestational age and birthweight were compared between both groups. Adjustments were made for maternal age, parity, cycle rank number, treatment indication, ART method, embryo characteristics and sex of the child. RESULTS: Singletons born after DET have a significantly lower birthweight than that after SET (3204.3 ± 617.5 g versus 3324.6 ± 509.7 g, P < 0.01). Also preterm birth (

Background The aim of our study was to review the results of microsurgically performed testicular sperm extraction (TESE) and to evaluate its possible long term effects on serum testosterone (T). Methods We operated on 48 men (35 +/- 8 years) with non-obstructive azoospermia (NOA). If no spermatozoa were found following a micro epididymal sperm extraction (Silber et al., 1994) and testicular biopsy, testicular microdissection was performed or multiple microsurgical testicular biopsies were taken. The mean follow-up of the serum T was 2.4 +/- 1.1 years. Results Sperm was retrieved in 17/48 (35%) of the men. The per couple take home baby rate if sperm was retrieved was 4/17 (24%). Serum T decreased significantly at follow-up (p < 0.05) and 5/31 (16%) de novo androgen deficiencies developed Conclusion In patients with non-obstructive azoospermia in whom no spermatozoa were found following a micro epididymal sperm aspiration and a simple testicular biopsy, we were able to retrieve spermatozoa in 35% of the men. The take home baby rate was 24% among couples with spermatozoa present upon TESE. De novo androgen deficiency occurred in 16% of the male patients following TESE indicating that, in men with NOA, long term hormonal follow up is recommended after TESE.

BACKGROUND: First-trimester bleeding is frequent in assisted reproductive technique (ART) pregnancies. It is unknown whether first-trimester bleeding, if not ending in a spontaneous abortion, negatively influences further pregnancy outcome in ART in singletons. METHODS: Data were obtained from our ART database (1993–2002), with 1432 singleton ongoing pregnancies being included in this study. The outcome measures—second-trimester and third-trimester bleeding, preterm contraction rates, pregnancy duration, birthweight, Caesarean section rates, intrauterine growth retardation (IUGR), preterm prelabour rupture of membranes (P-PROM), neonatal intensive care unit (NICU) admission and perinatal mortality—were compared in the groups with and without first-trimester bleeding. RESULTS: Significantly more singleton pregnancies resulted from a vanishing twin in the group with firsttrimester bleeding (8.7%) than in the controls (4.0%). A correlation was found between the incidence of first-trimester bleeding and the number of embryos transferred. First-trimester bleeding led to increased second-trimester [odds ratio (OR) = 4.56; confidence interval (CI) = 2.76–7.56] and third-trimester bleeding rates (OR = 2.85; CI = 1.42– 5.73), P-PROM (OR = 2.44; CI = 1.38–4.31), preterm contractions (OR = 2.27; CI = 1.48–3.47) and NICU admissions (OR = 1.75; CI = 1.21–2.54). First-trimester bleeding increased the risk for preterm birth (OR = 1.64; CI = 1.05–2.55) and extreme preterm birth (OR = 3.05; CI = 1.12–8.31). CONCLUSIONS: First-trimester bleeding in an ongoing singleton pregnancy following ART increases the risk for pregnancy complications. The association between firsttrimester bleeding, the number of embryos transferred and adverse pregnancy outcome provides a further argument in favour of single-embryo transfer.

Introduction Pregnancy outcome after IVF has been shown to be worse than after spontaneous conception. There is discussion as to whether this results from the technique itself or the patient characteristics. This study compares pregnancy outcome after IVF and intra-uterine inemination (IUI) in a matched patient group. Methods Data were obtained from our IVF and IUI databases (1997-2001). Matching was performed for maternal age, parity and plurality, and 126 IUI pregnancies were compared with 126 IVF pregnancies. Outcome variables were pregnancy duration, birth weight, Caesarean section rates, preterm contraction rates, neonatal intensive care unit admission, Apgar score, blood loss rates and maternal hypertension. Results None of the analysed parameters was statistically different between the groups. Conclusion This matched case-control study does not show different pregnancy outcomes after IVF and IUI. Since there is no reason to believe that the IUI technique in itself leads to an increased obstetric or neonatal risk, this study suggests that the worse pregnancy outcome after IVF as compared with spontaneous conceptions is due to the specific patient characteristics, rather than to the use of IVF itself.

Cryopreserved human primordial follicles grafted into immunodeficient hosts are susceptible to gonadotrophic stimulation but the optimal interval between grafting and stimulation has not been determined. The effect of stimulation with FSH at different time intervals after grafting was therefore investigated. Cryopreserved human ovarian cortical grafts from an androgen pre-treated female-to-male transsexual patient were transplanted into four groups of four non-obese, diabetic severe combined immune deficiency (NOD-SCID) mice. Ten, 12, 14 and 17 weeks after grafting, stimulation was started with daily intra-peritoneal injections of 5 IU of recombinant FSH for 14 days. Grafts were recovered and serially sectioned for counting the number of follicles and determining the different stages of development. Significantly more primary and secondary follicles were found in all stimulated groups compared with the non-stimulated control. This progression from the primordial to growing primary and secondary stages was most significant in the 14 weeks interval group. Formation of antral follicles was scarce and was only noticed in the 10 and 17 week groups. Cystic follicles and follicles with structural oocyte abnormalities were encountered in each group. Whether this phenomenon is related to the hormonal state of the tissue donor or to the cryopreservation and transplantation procedures is the subject of further investigation. It is concluded that the interval between grafting of ovarian tissue and the start of gonadotrophic stimulation may be important for an optimal follicular development after grafting.

Background Ovarian tissue cryopreservation and transplantation can be used to restore fertility to sterile females. A question that warrants further investigation is whether the follicular content is affected by the freeze-thawing and grafting procedure, and if so, to what extent and by what mechanism. Methods and results Intact newborn mouse ovaries were allografted under the kidney capsule or were cryopreserved by slow freezing with dimethylsulphoxide as the cryoprotectant prior to grafting. Estrogenic activity of ovariectomized recipient mice, as revealed by vaginal cytology, resumed after 11 days of transplantation. At 14 days after transplantation, ovarian grafts were recovered and processed histologically for follicle number counting. The follicular content of grafts of fresh ovaries was 58% of that from ovaries of age-matched 14 day old mice. In frozen-thawed ovarian grafts, the follicular content was only 9% lower than that of fresh grafted ovaries. Apoptosis of follicular cells was investigated by DNA nick end labelling. We observed a marked increase in the staining of fragmentation of DNA shortly after transplantation (2-12 h) of fresh newborn mouse ovaries. Conclusions The results of the present study indicate that transplantation rather than cryopreservation accounts for the major and early loss of primordial follicles in grafted newborn mouse ovaries.

Objective : To investigate follicle growth in fresh and cryopreserved human ovarian cortical grafts transplanted to immunodeficient mice. Study design : Fresh or frozen-thawed human ovarian cortex was grafted subcutaneously or under the kidney capsule of 43 mice (35 nude mice and eight SCID mice), 14 of which were non-stimulated controls, 21 injected intra-peritoneally with gonadotrophins during 2 weeks and eight injected during 3 months. Follicle count was compared by Chi-square. Results : Proportions of primordial follicles were significantly lower in grafts than in the tissue before transplantation in gonadotrophin-stimulated mice (37% versus 79%), but not in non-stimulated mice (51% versus 74%). Proportions of primary and secondary follicles were increased after transplantation indicating early follicular growth. One antral follicle was observed in a graft in a mouse stimulated for 3 months. Conclusion : Primordial follicles in fresh or frozen-thawed human ovarian cortex transplanted under the kidney capsule or subcutaneously can grow and are responsive to hormonal stimulation. Condensation : Primordial follicles in fresh and cryopreserved human ovarian cortical grafts can initiate growth after transplantation to immunodeficient mice

Along with improved survival, methods to preserve or restore the fertility potential of young women and children treated with cytotoxic chemotherapy or pelvic radiotherapy have been developed or are in the offing. Surgery, radiotherapy and chemotherapy can all impact on the future ovarian function, but patient and disease tailored application and use of preventive measures can limit ovarian damage. When the loss of reproductive ovarian function is unavoidable, different alternatives to preserve fertility or at least to restore the procreative potential are available. Creation of embryos by IVF, oocyte donation and cryopreservation of mature or immature oocytes are potential issues, the advantages and limitations of which are discussed. Recently, ovarian tissue cryopreservation has spurred interest in the medical literature as well as in the lay press as a method for preservation and restoring fertility. Considering the available data and current state of knowledge, we want to stress that this methodology is still in an experimental phase and we would like to caution against unwarranted enthusiasm of physicians and patients. The medical information preceding the informed consent should mention the actual uncertainties of this method. Moreover, the imperative character of the offer and in particular for paediatric oncological patients, the force of the moral rules that define parental obligations towards children should not be ignored.

Objective To investigate the optimal protocol for cryopreservation of human embryos obtained from IVF. Design Prospective randomized study. Setting Consenting patients in an academic research environment. Patients Couples undergoing IVF. Interventions A cohort of 2,220 supernumerary multicellular embryos were obtained from 488 patients who were randomized over slow freezing protocols with dimethylsulfoxide (DMSO, 819 embryos), 1,2-propanediol (699 embryos) or a mixture of DMSO and 1,2-propanediol (702 embryos). A total of 725 embryos have been thawed (DMSO, 232 embryos; 1,2-propanediol, 250 embryos and DMSO and 1,2-propanediol, 243 embryos) for transfer in natural ovarian cycles. Main Outcome Measures Embryo survival rate, embryo implantation rate, clinical pregnancy rate (PR), delivery rate, live-birth rate. Results The embryo survival rate was significantly higher with the DMSO protocol (52.6%) than with the 1,2-propanediol (32.0%) or the DMSO and 1,2-propanediol protocols (34.9%). The clinical PR per thawing cycle was significantly higher in the DMSO protocol (17.2%) than in the 1,2-propanediol protocol (3.9%). The clinical implantation rate per embryo thawed was significantly different between a DMSO-frozen embryo (4.7%) and a 1,2-propanediol-frozen embryo (1.2%). A DMSO and 1,2-propanediol-frozen embryo had a 3.7% chance of implantation. The delivery rate per thawing cycle was significantly higher in the DMSO protocol (12.5%) than in the 1,2-propanediol protocol (2.6%). The live-birth rates per embryo thawed were 3.5%, 0.8%, and 2.9% in the DMSO, 1,2-propanediol, and DMSO and 1,2-propanediol groups, respectively. Conclusion Supernumerary multicellular embryos as presented in daily clinical IVF practice have the highest chance of survival and of implantation after cryopreservation when DMSO has been used.

Liver metastases were produced in syngeneic BD IX rats by intraportal injection of colon cancer cell aggregates. The cells originated from the DHD/K12 cell line, derived from a 1,2-dimethylhydrazine (CAS: 540-73-8)-induced colon adenocarcinoma in BD IX rats. The animals received either cyclosporine A (CSA) or the excipients alone (control) through daily gastric intubation during 6 weeks. Multiple and very large hepatic metastases were observed early in 100% of the CSA-treated rats. The mean tumor volume was approximately 2,000 times higher in the CSA-treated group than in the controls (P less than .01). Survival time in the CSA-treated group was shortened (P less than .01) by generalized metastatic disease. Easy production of metastasis from colon cancer in 100% of the animals and precise estimation of tumor volume may prove useful for future therapeutic studies of secondary hepatic disease.