MYO10 promotes transzonal projection-dependent germ line-somatic contact during mammalian folliculogenesis†.

Granulosa cells of growing ovarian follicles elaborate filopodia-like structures termed transzonal projections (TZPs) that supply the enclosed oocyte with factors essential for its development. Little is known, however, of the mechanisms underlying the generation of TZPs. We show in mouse and human that filopodia, defined by an actin backbone, emerge from granulosa cells in early stage primary follicles and that actin-rich TZPs become detectable as soon as a space corresponding to the zona pellucida appears. mRNA encoding Myosin10 (MYO10), a motor protein that accumulates at the base and tips of filopodia and has been implicated in their initiation and elongation, is present in granulosa cells and oocytes of growing follicles. MYO10 protein accumulates in foci located mainly between the oocyte and innermost layer of granulosa cells, where it colocalizes with actin. In both mouse and human, the number of MYO10 foci increases as oocytes grow, corresponding to the increase in the number of actin-TZPs. RNAi-mediated depletion of MYO10 in cultured mouse granulosa cell-oocyte complexes is associated with a 52% reduction in the number of MYO10 foci and a 28% reduction in the number of actin-TZPs. Moreover, incubation of cumulus-oocyte complexes in the presence of epidermal growth factor, which triggers a 93% reduction in the number of actin-TZPs, is associated with a 55% reduction in the number of MYO10 foci. These results suggest that granulosa cells possess an ability to elaborate filopodia, which when directed toward the oocyte become actin-TZPs, and that MYO10 increases the efficiency of formation or maintenance of actin-TZPs.

Outcomes of Preimplantation Genetic Testing for Single Gene Defects in a Privately Funded Period and Publicly Funded Period: A North-American Single Center Experience.

BACKGROUND: The purpose of this study was to assess whether the outcomes from IVF-preimplantation genetic testing (IVF-PGT) cycles for single gene defects (SGD) (PGT-M) differ between a privately funded period (PRP) and publicly funded period (PUP). METHODS: A retrospective cohort study was conducted in a North-American single tertiary center. The PRP (March 1998 to July 2010) comprised 56 PGT-M cycles from 58 IVF cycles in 38 couples, and the PUP (August 2010 to May 2015) comprised 59 PGT-M cycles from 87 IVF cycles in 38 couples. One PGT-M cycle is defined as one biopsy procedure from one or serial IVF cycles. A p-value of 0.05 was considered statistically significant. RESULTS: The clinical pregnancy rates (CPR) per PGT-M cycle were 30.4% and 52.5% in each period, respectively (p=0.021). The live birth rates (LBR) per PGT-M cycle were 21.5% versus 40.9% in each period, respectively (p=0.037). A sub-analysis within the PUP comparing 39 PGT-M cycles from 39 IVF cycles with 20 PGT-M cycles from 49 IVF cycles yielded CPRs per PGT-M cycle of 64.1% and 30.0% and LBRs per PGT-M cycle of 53.8% and 15.0%, in each group, respectively (p< 0.05 for both). CONCLUSION: The transition from private to public funding and a single embryo transfer (ET) guideline has little impact on embryological and clinical outcomes of PGT-M cycles, and results in lower rates of multiple pregnancies. However, these two systems may serve different populations.

Human oocytes harboring damaged DNA can complete meiosis I.

OBJECTIVE: To determine whether human oocytes possess a checkpoint to prevent completion of meiosis I when DNA is damaged. DESIGN: DNA damage is considered a major threat to the establishment of healthy eggs and embryos. Recent studies found that mouse oocytes with damaged DNA can resume meiosis and undergo germinal vesicle breakdown (GVBD), but then arrest in metaphase of meiosis I in a process involving spindle assembly checkpoint (SAC) signaling. Such a mechanism could help prevent the generation of metaphase II (MII) eggs with damaged DNA. Here, we compared the impact of DNA-damaging agents with nondamaged control samples in mouse and human oocytes. SETTING: University-affiliated clinic and research center. PATIENT(S): Patients undergoing ICSI cycles donated GV-stage oocytes after informed consent; 149 human oocytes were collected over 2 years (from 50 patients aged 27-44 years). INTERVENTIONS(S): Mice and human oocytes were treated with DNA-damaging drugs. MAIN OUTCOME MEASURE(S): Oocytes were monitored to evaluate GVBD and polar body extrusion (PBE), in addition to DNA damage assessment with the use of γH2AX antibodies and confocal microscopy. RESULT(S): Whereas DNA damage in mouse oocytes delays or prevents oocyte maturation, most human oocytes harboring experimentally induced DNA damage progress through meiosis I and subsequently form an MII egg, revealing the absence of a DNA damage-induced SAC response. Analysis of the resulting MII eggs revealed damaged DNA and chaotic spindle apparatus, despite the oocyte appearing morphologically normal. CONCLUSION(S): Our data indicate that experimentally induced DNA damage does not prevent PBE in human oocytes and can persist in morphologically normal looking MII eggs.

Collection of 125 oocytes in an in vitro maturation cycle using a new oocyte collection technique: a case report.

BACKGROUND: We have developed a new oocyte collection technique applicable to use in women with "string-of-pearls" polycystic ovaries undergoing in vitro maturation (IVM) of oocytes for in vitro fertilization. CASE: A 34-year-old woman with polycystic ovary syndrome and infertility underwent IVM. Her ovaries had the string-of-pearls appearance on ultrasound, and antral follicle counts were consistently less than 60. An IVM cycle was performed using a new "rapid-pass" oocyte collection technique. We retrieved 125 germinal vesicle oocytes. A total of 44 oocytes reached the metaphase II stage after 48 hours in culture. After fertilization, four embryos were transferred to the uterus, resulting in a live birth. CONCLUSION: We believe this to be the largest number of oocytes retrieved from a single individual at one time. This was done using a newly developed aspiration technique.

Contexte : Nous avons mis au point une nouvelle technique de collecte d'ovocytes pouvant être utilisée chez les femmes qui présentent des ovaires polykystiques « en collier de perles ¼ (string-of-pearls) et pour lesquelles ces ovocytes seront soumis à une maturation in vitro (MIV) aux fins de la fécondation in vitro. Cas : Une femme de 34 ans aux prises avec le syndrome des ovaires polykystiques et l'infertilité a eu recours à la MIV. L'apparence en collier de perles de ses ovaires a été révélée par l'échographie; de plus, la numération des follicules antraux était régulièrement inférieure à 60. Un cycle de MIV a été mené par l'intermédiaire d'une nouvelle technique « rapide ¼ de collecte d'ovocytes. Nous avons récupéré 125 ovocytes (vésicules germinatives). Au total, 44 ovocytes ont atteint le stade de la métaphase II après 48 heures en culture. À la suite de la fécondation, quatre embryons ont été transférés dans l'utérus, le tout s'étant soldé par une naissance vivante. Conclusion : Nous estimons qu'il s'agit là du plus grand nombre d'ovocytes récupérés chez une seule et même femme, au même moment. Nous y sommes parvenus au moyen d'une technique d'aspiration nouvellement conçue.

Live birth following serial vitrification of embryos and PGD for fragile X syndrome in a patient with the premutation and decreased ovarian reserve.

PURPOSE: To present a live birth resulting from serial vitrification of embryos and pre-implantation genetic diagnosis (PGD). METHODS: A 31-year-old with primary infertility, fragile-X premutation, and decreased ovarian reserve (DOR) (baseline FSH level 33 IU/L), presented after failing to stimulate to follicle diameters >10 mm with three cycles of invitro fertilization (IVF). After counseling, the couple opted for serial in-vitro maturation (IVM), embryo vitrification, and genetic testing using array comparative genomic hybridization (aCGH) and PGD. Embryos were vitrified 2 days after intra-cytoplasmic sperm injection (ICSI). Thawed embryos were biopsied on day-three and transferred on day-five. RESULTS: The couple underwent 20 cycles of assisted reproductive technology. A total of 23 in-vivo mature and five immature oocytes were retrieved, of which one matured in-vitro. Of 24 embryos, 17/24 (71 %) developed to day two and 11/24 (46 %) survived to blastocyst stage with a biopsy result available. Four blastocysts had normal PGD and aCGH results. Both single embryo transfers resulted in a successful implantation, one a blighted ovum and the other in a live birth. CONCLUSIONS: Young patients with DOR have potential for live birth as long as oocytes can be obtained and embryos created. Serial vitrification may be the mechanism of choice in these patients when PGD is needed.

Case report: the use of annexin V coupled with magnetic activated cell sorting in cryopreserved spermatozoa from a male cancer survivor: healthy twin newborns after two previous ICSI failures.

PURPOSE: The aim of the study is to report successful outcome (live births) after sperm sorting with annexin V-MACS on cryopreserved spermatozoa with high level of sperm DNA fragmentation from a cancer patient survivor. METHODS: Cryopreserved spermatozoa were sorted with annexin V-MACS prior to ICSI. Sperm DNA fragmentation was evaluated by SCSA(®) and TUNEL. RESULTS: The couple had two previous IVF/ICSI cycles failures using sperm cryopreserved before cancer treatment. On third ICSI cycle attempt results were as follow: pre-annexin V-MACS sperm quality: 10 × 10(6)/ml, 3.3 % progressive motility, 1 % normal forms, TUNEL: 72.5 % positive cells, SCSA(®): 76.6 % DFI. Post-annexin V-MACS sperm quality: 2.8 × 10(6)/ml, 10 % progressive motility, TUNEL: 58.8 % positive cells. Eight metaphase II oocytes were collected, 4 fertilized, 2 embryos were transferred on day 3 and healthy twins were born (1 boy, 1 girl). CONCLUSIONS: Annexin V-MACS technique could be a potential tool to improve sperm quality on cryopreserved spermatozoa of cancer patient and improve ICSI outcome.

Fertilization and embryo development with spermatozoa obtained from testicular sperm extraction into oocytes generated from human chorionic gonadotropin-primed in vitro maturation cycles.

OBJECTIVE: To evaluate the fertilization rate and embryo development resulting from intracytoplasmic sperm injection (ICSI) of spermatozoa retrieved by testicular sperm extraction (TESE) in hCG-primed in vitro maturation (IVM) cycles. DESIGN: Case-control study. SETTING: University teaching hospital. PATIENT(S): Twenty-four IVM cycles were performed in 21 patients (mean age, 32.3 ± 2.4 years) with polycystic ovaries (PCO) whose partners were nonobstructive azoospermic. Twelve cycles where IVM oocytes were also retrieved were compared with a control group consisting of age-matched IVM cycles with ICSI using ejaculated spermatozoa (n = 12). INTERVENTION(S): In vitro maturation treatment with TESE sperm. MAIN OUTCOME MEASURE(S): Fertilization and embryo development between sibling oocytes matured in vivo and in vitro. RESULT(S): Eight singleton pregnancies and one twin pregnancy were obtained after ET (9/24, 37.5%). In the 12 IVM cycles where in vivo-matured oocytes were also obtained, the fertilization rate after TESE-ICSI was significantly higher in in vivo-matured oocytes than in sibling in vitro-matured oocytes (84.2% vs. 53.2%). The proportion of good quality embryos was also higher (63.5% vs. 40.2%). In the control group of cycles with ejaculated spermatozoa, there was no difference in fertilization rates between sibling oocytes matured in vivo and in vitro (84.6% vs. 79.6%). CONCLUSION(S): Our results suggest that IVM of immature oocytes combined with TESE-ICSI is an option for couples with PCO and azoospermia. However, there are lower fertilization and good quality embryo rates achieved when TESE-ICSI was done with in vitro-matured oocytes. Additional studies are necessary to determine the role of this treatment combination.

Fertilization, embryo development, and clinical outcome of immature oocytes obtained from natural cycle in vitro fertilization.

PURPOSE: To analyze the fertilization, embryo development, and clinical outcome of immature oocytes obtained from natural cycle IVF in women with regular cycles. METHODS: Natural cycle IVF was performed in 28 patients who had normal ovaries, > 6 antral follicle counts and were less than 40 years old (n=28 cycles). An hCG trigger of 10,000 IU was administered 36 h before oocyte collection when the diameter of the dominant follicle (DF) was over 12 mm. Oocytes were retrieved from DF as well as from the cohort of smaller follicles. Embryological aspects of the mature and immature oocytes retrieved from these cycles as well as the implantation and clinical pregnancy rates depending on the origin of the embryos transferred were evaluated. RESULT(S): Overall clinical pregnancy and implantation rates were 20.8 % and 6.7 %, respectively. There were no differences in in vitro maturation (IVM), fertilization and embryo development between immature oocytes retrieved with and without in vivo matured oocytes. However, the clinical and implantation rates in cycles with embryos produced from in vivo matured oocytes transferred were better than the cycles where only IVM embryos were transferred (30.8 %, 9.1 % vs. 9.1 %, 3.2 %). CONCLUSION(S): Although our results show that immature oocytes from natural cycle IVF can fertilize normally and can be used to increase the number of embryos available for transfer, the embryos derived from the immature oocytes in natural cycles IVF have a poorer reproductive potential.

Chromosome segregation analysis in human embryos obtained from couples involving male carriers of reciprocal or Robertsonian translocation.

The objective of this study was to investigate the frequency and type of chromosome segregation patterns in cleavage stage embryos obtained from male carriers of Robertsonian (ROB) and reciprocal (REC) translocations undergoing preimplantation genetic diagnosis (PGD) at our reproductive center. We used FISH to analyze chromosome segregation in 308 day 3 cleavage stage embryos obtained from 26 patients. The percentage of embryos consistent with normal or balanced segregation (55.1% vs. 27.1%) and clinical pregnancy (62.5% vs. 19.2%) rates were higher in ROB than the REC translocation carriers. Involvement of non-acrocentric chromosome(s) or terminal breakpoint(s) in reciprocal translocations was associated with an increase in the percent of embryos consistent with adjacent 1 but with a decrease in 3∶1 segregation. Similar results were obtained in the analysis of nontransferred embryos donated for research. 3∶1 segregation was the most frequent segregation type in both day 3 (31%) and spare (35%) embryos obtained from carriers of t(11;22)(q23;q11), the only non-random REC with the same breakpoint reported in a large number of unrelated families mainly identified by the birth of a child with derivative chromosome 22. These results suggest that chromosome segregation patterns in day 3 and nontransferred embryos obtained from male translocation carriers vary with the type of translocation and involvement of acrocentric chromosome(s) or terminal breakpoint(s). These results should be helpful in estimating reproductive success in translocation carriers undergoing PGD.

Normal birth following PGD for reciprocal translocation after serial vitrification of oocytes from a poor responder: a case report.

This case study reports the first successful birth outcome following preimplantation genetic diagnosis (PGD) for a chromosome translocation in embryos generated by serial vitrification of oocytes. A couple presented to the fertility clinic with 2 years of primary infertility. The woman was diagnosed with poor ovarian reserve and her partner was diagnosed with severe oligoteratozoospermia and the reciprocal translocation 46,XY,t(1;7)(p36.1;q11.23). Following counselling, the couple opted for serial vitrification of oocytes followed by PGD. A total of 31 oocytes were obtained in five egg collection cycles over a period of 12 months and 27 metaphase-II oocytes were vitrified. Nineteen of the 27 vitrified oocytes survived warming: 14 oocytes from the vitrified group and three oocytes from the fresh cycle were fertilized by intracytoplasmic sperm injection. Eleven embryos, including three from the fresh cycle, were biopsied on day 3 post insemination. Fluorescence in-situ hybridization was performed for the specific chromosomes involved in translocation. Only two embryos from the cryopreservation cycles were diagnosed as normal/balanced, one of which was transferred on day 5 post insemination. A normal healthy female infant was born at week 42 of gestation.

Live birth after ICSI of micro-TESE-retrieved spermatozoa into in vitro-matured oocytes.

This case report describes a live birth resulting from intracytoplasmic sperm injection (ICSI) of spermatozoa retrieved by microdissection testicular sperm extraction (micro-TESE) into oocytes produced from human chorionic gonadotropin-primed in vitro maturation (IVM) cycles. In the IVM treatment, a total of 30 oocytes (1 mature and 29 immature oocytes) were retrieved. Following IVM, 9 oocytes had matured. A total of 4 oocytes were fertilized after ICSI with the husband's micro-TESE spermatozoa and 4 embryos were transferred into the uterus on day 3. A healthy boy weighing 2500 g was born at 35.5 weeks of gestation.

Comparison of fertilization and embryonic development in sibling in vivo matured oocytes retrieved from different sizes follicles from in vitro maturation cycles.

PURPOSE: To examine the fertilization and developmental potential of sibling mature oocytes collected from different follicle sizes on day of retrieval in in vitro maturation (IVM) cycles. METHODS: Two hundred thirty eight hCG-primed IVM cycles were performed in 213 patients with polycystic ovaries. If sibling mature oocytes were retrieved on day of collection, they were divided into two groups, Group 1 (n = 78): M-II oocytes obtained from follicles size 10-14 mm; Group 2 (n = 192): M-II oocytes obtained from follicles size <10 mm. RESULTS: Of the 238 cycles, 63 cycles had more than one M-II oocytes retrieved (total M-II oocytes = 270) both from Groups 1 and 2. There were no significant differences between the two groups for oocyte diameter (117.2 mm vs. 116.9 mm), fertilization (79.5% vs. 72.4%) and good quality embryo on day 3 (66.1% vs. 56.8%). CONCLUSIONS: The M-II oocytes retrieved from the cohort of follicles (<10 mm diameter) can produce the same quality of embryos as that from large follicles, likely contributing to improve the clinical outcome.

Comparison of survival rate of cleavage stage embryos produced from in vitro maturation cycles after slow freezing and after vitrification.

Significantly more embryos survived the vitrification procedure compared to slow freezing (85.5% vs. 61.8%) in cleavage-stage human embryos produced from in vitro maturation cycles, suggesting that vitrification is more efficient than slow freezing for cryopreservation.

Selection of the optimal day for oocyte retrieval based on the diameter of the dominant follicle in hCG-primed in vitro maturation cycles.

BACKGROUND: The efficiency of in vitro maturation (IVM) techniques is suboptimal compared with controlled ovarian stimulation combined with IVF cycles, and studies are needed to identify factors that predispose IVM cycles to success or failure. We compared the outcome of IVM cycles with different dominant follicle (DF) size at oocyte retrieval following hCG priming. METHODS: IVM was performed in 160 patients with polycystic ovaries (171 cycles). We administered 10,000 IU hCG s.c. 35-38 h before oocyte collection when endometrial thickness reached at least 6 mm. IVM cycles were retrospectively analyzed according to DF diameter as follows; Group 1: DF diameter 14 mm. RESULTS A positive correlation was observed between DF size and number of in vivo matured oocytes collected (Group 1, 2 and 3 = 6.9, 10.6 and 15.1%, respectively). The rates of IVM, fertilization and embryo development were similar among the sibling immature oocytes collected from the three groups. However, clinical pregnancy rate in Group 2 (40.3%) was higher than Group 3 (17.1%) (P < 0.05). Moreover, implantation rates in Groups 1 (13.6%) and 2 (14.3%) were higher than Group 3 (4.9%) (P < 0.01). CONCLUSIONS: Our results suggest that oocyte collection in IVM cycles should be performed when the DF is 14 mm diameter or less. Sibling immature oocytes may be affected detrimentally if a DF >14 mm is present at oocyte collection.

Comparison of in-vitro maturation cycles with and without in-vivo matured oocytes retrieved.

This study compared the embryological characteristics and clinical outcome of in-vitro maturation (IVM) treatment cycles with and without in-vivo matured oocytes collected following human chorionic gonadotrophin (HCG) priming. The patients were administered 10,000 IU of HCG subcutaneously when endometrial thickness reached > or =6 mm and oocyte collection was performed 35-36 h after HCG administration. The clinical outcome and embryological aspects were analysed between IVM cycles with (group 1) and without (group 2) in-vivo matured oocytes. In group 1, three (range 1-12) in-vivo matured oocytes per patient were retrieved on average. The number of good quality embryos derived from in-vivo matured oocytes in group 1 was significantly higher than those derived from in-vitro matured oocytes in group 1 and group 2 (P < 0.05). However, there was no difference between the number of good quality embryos produced from in-vitro matured oocytes in the two groups. There were 12 clinical pregnancies (40.0%) in group 1, and seven pregnancies (23.3%) in group 2. These results suggest that IVM cycles with in-vivo matured oocytes resulted in a good clinical pregnancy rate, which could be explained by the superior quality of embryos derived from the in-vivo matured oocytes.

A 38 h interval between hCG priming and oocyte retrieval increases in vivo and in vitro oocyte maturation rate in programmed IVM cycles.

BACKGROUND: Our aim was to evaluate whether extending the interval between human chorionic gonadotrophin (hCG) priming and immature oocyte retrieval increases the oocyte maturation rate following in vitro maturation (IVM). METHODS: This study was performed retrospectively. IVM was performed on 113 polycystic ovary syndrome patients (n = 120 cycles). Oocyte collection was performed either 35 h (Group 1; n = 76) or 38 h (Group 2; n = 44) after 10,000 IU of hCG priming. Following oocyte retrieval, oocyte maturity was assessed and the remaining immature oocytes were cultured in IVM medium up to Day 2. RESULTS: The number of in vivo matured oocytes collected was significantly higher in Group 2 (13.6%, 114/840 versus 7.3%, 96/1312 in Group 1) (P < 0.01); the oocyte maturation rate after Day 1 was significantly higher (P < 0.01) in Group 2 (46.3 versus 36.0% in Group 1); and clinical pregnancy (40.9 versus 25%) and implantation rates (15.6 versus 9.6%) were better in Group 2 than those in Group 1. CONCLUSIONS: The results suggest that extending the period of hCG priming time from 35 to 38 h for immature oocyte retrieval promotes oocyte maturation in vivo and increases the IVM rate of immature oocytes. Therefore, oocyte retrieval after 38 h of hCG priming may improve subsequent pregnancy outcome in cycles programmed for IVM treatment.

Effect of polyvinylpyrrolidone on bovine oocyte maturation in vitro and subsequent fertilization and embryonic development.

The exact role of polyvinylpyrrolidone (PVP) in culture medium for oocyte maturation is still largely unknown. Bovine cumulus-oocyte complexes (COC) were cultured in in-vitro maturation (IVM) medium supplemented with 10% fetal bovine serum (FBS), 0.3% PVP (K-90) or 10% serum substitute supplement (SSS) respectively. The rates of oocyte maturation, fertilization and early embryonic development were evaluated. In addition, the status of DNA fragmentation in the oocytes was determined by comet assay, and the ratio of trophectoderm (TE) cells and inner cell mass (ICM) in blastocysts was determined by differential staining. Furthermore, the percentage of apoptotic cells in the blastocysts was examined by TUNEL assay. The results indicated that the effect of PVP in IVM medium was similar to FBS in terms of oocyte maturation and subsequent embryonic development. However, the addition of SSS in IVM medium retarded further embryonic development and resulted in more oocyte DNA fragmentation and a higher ratio of TE cells and ICM in the blastocysts. However, the number of apoptotic cells in blastocysts was similar among the three groups. These results suggest for the first time that the addition of PVP in oocyte maturation medium is not only a suitable substitute for serum but is also beneficial to in-vitro oocyte maturation.

High survival and hatching rates following vitrification of embryos at blastocyst stage: a bovine model study.

Cryopreservation of embryos at the blastocyst stage may provide an effective method to increase the cumulative pregnancy rate for each treatment cycle of ovarian-stimulated IVF. The objective of this study was to evaluate the survival rate and hatching rate of bovine blastocysts following vitrification using a method designed for oocytes, with a view to introducing this methodology into human assisted reproduction technology and reproductive medicine. Bovine blastocysts were produced from abattoir materials subjected to in-vitro maturation and in-vitro fertilization. Survival rate of the bovine blastocysts was 100% (94/94) following vitrification using a method designed for oocyte cryopreservation. There was no difference in the hatching rate of the bovine blastocysts between control (62.5%: 60/96) and vitrified (61.7%: 58/94) groups. The number of dead cells in the blastocysts was not significantly different between control (5.0 +/- 2.9) and vitrified (9.5 +/- 4.0) groups. In conclusion, the results of this study indicate that bovine blastocysts can be vitrified successfully using a procedure designed for oocyte cryopreservation. It is possible that this method may also be successful for the cryopreservation of human embryos. A further study into this is currently being organized.

[Accuracy of endoscopic ultrasonographic impression compared with pathologic diagnosis in gastrointestinal submucosal tumors].

BACKGROUND/AIMS: Endoscopic ultrasonography (EUS) is a valuable imaging modality for the evaluation of gastrointestinal submucosal tumor (SMT). EUS is helpful in assessing the layer of origin, tumor diameter, shape, border characteristics, and internal echo patterns of SMTs and thus makes it possible to predict histologic diagnosis with educated guess. However, some studies have found no significant differences in EUS features between benign and malignant mesenchymal tumors. By comparing EUS impressions with histologic diagnosis, we evaluated the accuracy of EUS in differential diagnosis of gastrointestinal SMTs. METHODS: 58 cases of gastrointestinal SMTs with both EUS findings and pathologic reports were compared retrospectively from August 2001 to September 2003. RESULTS: 34 patients had lesions in the stomach and 13, 8, 3 in the esophagus, duodenum, and colon respectively. Benign lesions were predominant (46 of 58). The EUS and pathologic diagnosis coincided in 46/58 (79.3%) of the cases. Use of EUS led to the correct diagnosis in 7/9 (77.8%) of malignant GISTs (gastrointestinal stromal tumor) and leiomyosarcomas. Two small malignant gastric GISTs were diagnosed as benign with EUS. CONCLUSIONS: EUS is a useful tool in the differential diagnosis of gastrointestinal SMTs and predicting malignant lesions. However, some malignant GISTs were diagnosed as benign tumor with EUS examination.

Maturational and developmental competence of immature oocytes retrieved from bovine ovaries at different phases of folliculogenesis.

Immature oocytes recovered from bovine ovaries were studied to determine if their maturational and developmental competence is affected by phase of folliculogenesis. Ovaries (a total of 39 pairs) were collected from a local abattoir. Following examination, each pair of ovaries was assigned to one of three groups, according to follicle size and with or without a corpus luteum: (i) early phase (n = 13 pairs): all follicles were or=15 mm in diameter; (iii) luteal phase (n = 13 pairs): all follicles were