Comparison of different methods for the recovery of horse oocytes

The object of this study was to compare 4 different methods of oocyte recovery from mares; 1) transvaginal follicle aspiration in vivo; 2) follicle aspiration in vitro; 3) oocyte recovery by isolation of follicles in vitro and 4) follicle scraping in vitro. Oocyte recovery was highest after follicle scraping (71.1%) and follicle isolation and rupture (61.3%). Follicle aspiration in vitro and in vivo yielded oocytes on 31.2% and 19.3% of occasions, respectively. The output of different types of cumulus-oocyte-complexes was different among the methods; the portion of compact cumulus-oocyte-complexes was significantly higher with follicle scraping (50.7%) and follicle isolation (44.5%) than with aspiration in vivo (31.9%) and in vitro (23.7%). The recovery rate of oocytes from small follicles (<15 mm) was significantly higher than from larger follicles (P<0.05) using transvaginal follicle aspiration. The proportion of oocytes that were degenerate (exhibited shrunken, dense or visibly damaged ooplasm) ranged from 1.2% after follicle scraping, to 17.2% after aspiration in vivo. These results indicate that, for the recovery of horse oocytes in vitro, follicle scraping and follicle isolation give the highest recoveries of cumulus-intact oocytes.     

The correlation between follicular measurements, oocyte morphology, and fertilization rates in an in vitro fertilization program

OBJECTIVE: To explore the relationship between follicle size and the morphology of the oocyte-cumulus-corona complex with fertilization rates in stimulated cycles of IVF. DESIGN: Retrospective comparison of measurements and observations of 2,429 oocytes from 215 patients undergoing 324 stimulated IVF cycles. SETTING: A large hospital-based IVF program. MAIN OUTCOME MEASURES: Individual follicles were measured by ultrasound before transvaginal aspiration and the size was recorded. The oocyte-cumulus-corona complex from each follicle was examined and classified. The oocytes were checked for evidence of fertilization 17 to 22 hours after insemination. RESULTS: The fertilization rate of all oocytes regardless of morphological type revealed a positive linear correlation with increasing follicle diameter. The fertilization rates of type I oocytes was marginally higher than type II oocytes, controlling for follicle diameter; however, this difference did not achieve statistical significance. Oocytes from follicles with a mean diameter > or = 16 mm had significantly higher fertilization rates than did oocytes from follicles with a mean diameter < or = 14 mm. CONCLUSIONS: Follicle size is a better predictor of fertilization than is morphological characterization of the oocyte-cumulus-corona complex in IVF.     

Clinical application of volume estimation based on three-dimensional ultrasonography

The development of three-dimensional sonography represents a milestone in the history of diagnostic ultrasound. It has become possible to evaluate a region of interest not only in the horizontal and vertical planes but also in a third, i.e. the frontal, plane. Among other possible applications, this technique greatly facilitates volume estimation. The objective of the present study was to evaluate the accuracy and precision of volumetry based on three-dimensional sonography. The results demonstrate a good correlation between estimated volumes and the volumes measured by transvaginal needle-guided follicle aspiration performed under sonographic visualization. Our findings confirm the conclusion of previous studies that three-dimensional sonography is making a valuable contribution to imaging techniques.     

Superovulatory responses in Japanese black beef cows following largest follicle aspiration or human chorionic gonadotrophin (hCG) treatment

The effect of largest follicle aspiration or hCG administration before induction of superovulation on the ovarian response of Japanese Black beef cows was investigated using a crossover design in which induction of superovulation was attempted in every cow. The superovulatory response of cows whose largest follicle had been aspirated from the ovaries by ultrasound-guided follicular aspiration 1 day before induction of superovulation, did not differ from the response in non-treated control cows. In contrast, in cows given 5,000 IU of hCG 3 days before induction of superovulation, the proportions of fertilized ova and transferable embryos significantly decreased compared with the other groups.     

Possibilities of ovum pickup in cattle

Cattle breeding has a new reproduction technique in addition to artificial insemination and embryo transfer. It is the collection of ova from living animal by means of ultrasound guided follicle aspiration (ovum pick-up), followed by embryo-production in vitro. Follicles larger than 2 mm were punctured and the ova were collected twice weekly during 3 months. In total 1677 ova were collected from ten cows; 1342 (80%) were used for in vitro maturation, fertilization, and embryoculture. All ova were fertilized with semen from one bull, and 218 transferable embryos were produced. Calculated on a year basis, this would amount to 87 embryos per animal, with an intra-animal variation between 28 and 132. This new technique may replace MOET (Multiple Ovulation and Embryo Transfer; yearly average of 25 transferable embryos per animal), if the embryo-production via OPU can be performed with semen from any selected bull.     

Follicle aspiration, sperm injection, and assisted rupture (FASIAR): a simple new assisted reproductive technique

OBJECTIVE: To describe the first successful application of a new fertility-enhancing technique. DESIGN: Case report. SETTING: Academic fertility program. PATIENT(S): A 36-year-old nulligravid woman undergoing inseminations with frozen-thawed donor sperm. INTERVENTION(S): Ovarian superovulation, follicle aspiration, sperm injection, and assisted follicular rupture. MAIN OUTCOME MEASURE(S): Assessment of feasibility of technique and pregnancy outcome. RESULT(S): After failing to conceive during 16 cycles of IUI, the patient successfully achieved an ongoing pregnancy during the second follicle aspiration, sperm injection, and assisted rupture (FASIAR) attempt. CONCLUSION(S): Follicle aspiration, sperm injection, and assisted rupture combines the concepts of superovulation, IUI, and peritoneal oocyte and sperm transfer to obviate the possibility of luteinized unruptured follicle syndrome, assist oocyte release, and ensure gamete intermixing. It also can be used to reduce the number of ovulating oocytes and thus to reduce the risk of multiple gestations. Follicle aspiration, sperm injection, and assisted rupture is a new, simple, office-based procedure that does not require embryologic expertise beyond sperm preparation as for IUI, yet promises to be more successful than IUI.     

Ultrasound in an in vitro fertilization program

Ultrasound examinations of the preovulatory follicle were performed on 39 patients in 58 consecutive spontaneous cycles in which ovum aspiration for in vitro fertilization was planned. Examinations during the follicular phase helped to indicate when patients should be admitted for intensive monitoring of urinary luteinizing hormone (LH) levels and as a means of lateralizing the side of follicular development in those patients in whom one ovary was known to be inaccessible to laparoscopic aspiration. The technique was also of value in determining whether ovulation had occurred in those patients in whom the anticipated midcycle LH surge was not detected and as a routine measure prior to laparoscopy to ensure the continuing presence of the follicle.     

Possibilities of the use of 3-dimensional ultrasonography in gynecology and obstetrics

The authors report their experiences with 3 dimensional ultrasound technique applied in cases of 286 obstetrical and 35 gynecological examinations. Combison 530, 3 D ultrasound equipment was used. Pathological signs were found in 14 cases. The introduction of this technique has been increasing the diagnostic accuracy of prenatal and gynecological ultrasound examinations, and some malformations, that could have never been seen by "traditional" 2 D methods, can be visualized as well.     

Selection, dominance and atresia of follicles during the oestrous cycle of heifers

This study examined the correlation between measurement of follicle growth by ultrasound, and measurement of intrafollicular ratios of oestradiol and progesterone concentrations and the serum concentrations of FSH during selection, dominance and atresia or ovulation of dominant follicles in heifers. Heifers were ovariectomized on days 0 (before LH surge), 1 (after LH surge, preovulation), 1 (postovulation), 3, 6 and 12 of the oestrous cycle. Blood samples were collected at 4-6 h intervals. After ovariectomy all follicles > or = 5 mm were measured and follicular fluid was aspirated. Follicles were classified by size according to ultrasound (F1, largest; F2, second largest; F3, all remaining follicles > or = 5 mm) and by the ratio of oestradiol:progesterone concentrations. During the follicular phase, a single dominant oestrogen-active follicle increased in diameter while serum concentrations of LH increased and FSH decreased (P < 0.05). On day 1 (after LH surge, preovulation), serum LH and FSH decreased to pre-surge concentrations (P < 0.0001), while follicle size and intrafollicular progesterone concentration increased and oestradiol concentration decreased (P < 0.05). A dominant nonovulatory follicle, classified as oestrogen-active on days 1, 3 and 6 and oestrogen-inactive on day 12, increased in size from day 1 to day 7 and lost dominance during days 10-12, coincident with the growth of multiple oestrogen-active follicles. The serum FSH concentration increased transiently (P < 0.05) before each new wave of dominant follicular growth. The overall correlation of ultrasound measurements of follicle diameter with measures of follicle size after ovariectomy was high.(ABSTRACT TRUNCATED AT 250 WORDS)     

The new temperance movement in the United States: what happened to macrostructural factors in alcohol problems?

BACKGROUND: Pulmonary tuberculosis can produce unusual radiographic appearances and negative results of sputum and bronchoscopic examinations are common. This study assessed the value of ultrasound guided aspiration biopsy in the diagnosis of pulmonary tuberculosis with unusual radiographic appearances. METHODS: Thirteen patients, ultimately diagnosed as having tuberculosis, underwent a chest ultrasonographic examination between June 1984 and August 1991. All had sputum available for examination and nine were also examined by bronchoscopy. Ten patients who had a negative sputum smear and negative bronchoscopic brushing smears underwent ultrasound guided aspiration or biopsy. Percutaneous aspiration was performed with a 22 gauge needle. If the smear did not reveal acid fast bacilli, a biopsy sample was taken with a 16 gauge Tru-cut needle to obtain a histological diagnosis. RESULTS: The ultrasonographic examination delineated the more complex nature of the lesions better than the chest radiograph. Ultrasound guided aspiration biopsy provided the diagnosis in nine of 10 patients, while the sputum smear and culture provided diagnosis in five of 13, and bronchoscopy in four of nine. In terms of rapid diagnosis, ultrasound guided aspiration biopsy gave the diagnosis in eight of 10 cases. No patient developed a major complication. CONCLUSION: Ultrasonography can direct the needle to the most suitable part of a lesion to obtain the relevant specimens. The diagnostic yield is high and the procedure is relatively safe. It is especially helpful in patients with negative results of sputum and bronchoscopic examinations.     

Potentialities of ultrasound examination in differential diagnosis of benign nodules and breast cancer

The results of examination of 410 patients with nodular masses of the breast are analysed. Benign tumors were present in 314 (76.6%) of the patients, cancer of the mammary gland--in 96 (23.4%) of the patients. The tumors were classified into 3 types, depending on the intensity of the ultrasound wave behind the tumor. Based on the clinical and morphological analyses it was demonstrated that the presence of the acoustic wave behind the tumor is typical for the invasive cancer of the duct, scirrous carcinoma and lobular cancer. In medullar and mucosous cancer the intensity of the ultrasound wave does not change. In case of intracystic cancer the symptom of "back magnification" is present. The sensitivity of the ultrasound in detection of mammary gland cancer is 88.5%, specificity--96.5%. The analogous parameters of mammography are: 91.0% and 97.8%. In the use of ultrasound control in fine needle aspiration biopsy (FNAB) the sensitivity and specificity of the method increases from 88.5% to 92.7%. If the diagnosis of mammary gland cancer was established with the use of ultrasound and FNAB the additional use of mammography is not obligatory.     

Mammographically negative breast cancer at the Strathfield Breast Centre

BACKGROUND: The current diagnostic modalities used to detect breast cancer are mammography, together with clinical examination, ultrasound and fine needle aspiration biopsy (FNAB). The accuracy rates for each modality varies and a combination of the modalities is recommended to detect cancer early. Some authors have suggested that mammography should be used primarily as a screening tool because of the false negative mammography results that have been reported in the past 10 years. The records of patients at the Strathfield Breast Centre were reviewed to determine the accuracy of other modalities. METHODS: The records of 371 breast cancer patients treated at the Strathfield Breast Centre in the 6 years form 1989 to 1994 were reviewed to determine the accuracy of mammography, ultrasound, clinical examination and fine needle aspiration biopsy. Of the 371 women with histopathologically diagnosed breast cancer, 349 had mammography. RESULTS: The accuracy rate of mammography in the present study was 91% with a false negative rate of 9%. It was found that there was no significant delay in treatment of breast cancer in mammogram-negative patients. CONCLUSIONS: Mammographically negative breast cancer was found to be more common in younger women, to be similar in size to mammogram-positive cancer, to occur in all histological types and grades and was usually invasive rather than noninvasive. The rate of lymph node involvement was similar to the mammogram-positive group.     

Effects of follicular aspiration and flushing, and the genotype of the fetus on circulating progesterone levels during pregnancy in the mare

When aspirating ovarian follicles in pregnant mares to obtain oocytes for in vitro fertilisation (IVF), the effect of the manipulation on circulating concentrations of progesterone may be an important consideration in terms of the maintenance of pregnancy. The object of this study was to compare the effects of 3 different forms of transvaginal ultrasound-guided follicle aspiration (Treatment 1, no aspiration, n = 4; Treatment 2, aspirate only follicles > or =20 mm in diameter, n = 7; Treatment 3, aspirate all visible follicles, n = 7) on peripheral plasma progesterone concentrations between Days 21 and 150 of gestation in 9 mares carrying intraspecies horse and 9 mares carrying interspecies mule conceptuses. The 3 follicle aspiration treatments were applied at the peak of each follicular wave as determined by follicular mapping by means of transrectal ultrasonography on alternate days. The plasma progesterone profile in mares undergoing Treatment 1 was in close agreement with those reported previously in pregnant mares. A decline in plasma progesterone levels occurred after Day 53 of gestation in Treatments 2 and 3 mares, indicating that the follicular aspiration procedures did interfere with the formation of secondary corpora lutea. However, the levels in individual mares never dropped low enough to endanger the pregnancy. Mares carrying mule pregnancies exhibited higher mean plasma progesterone concentrations between Days 39 and 45 of gestation than mares carrying horse pregnancies, equivalent levels between Days 46 and 66 despite the lower circulating concentrations of chorionic gonadotrophin (mule CG) in their blood during this period and lower progesterone levels between Days 67 and 150 of gestation. The results indicate that the primary corpus luteum in the pregnant mare may be more sensitive to mule CG than horse CG. Furthermore, the earlier disappearance of CG from the circulation in mares carrying mule fetuses is reflected by an earlier decline in plasma progesterone concentrations in this type of equine pregnancy.     

Dietary fat and body composition

Vascular endothelial growth factor (VEGF) is a cytokine that induces angiogenesis. Angiogenesis is a prominent histologic component of the luteinization process. Luteinization is also characterized by granulosa cell progesterone secretion in response to the luteinizing hormone (LH) surge. Local VEGF production in human pre-ovulatory follicles, induced by LH, was postulated to be a luteinization mediator in women. To investigate this hypothesis, serum and fluid from the dominant follicle of 31 healthy regularly cycling multiparous women undergoing laparoscopic sterilization were obtained. VEGF was measured by enzyme-linked immunosorbent assay, and LH and progesterone were measured by radioimmunoassay. Follicle aspiration was performed at a median of 13 days from the last menstrual period (range 11-17 days). The median pre-ovulatory follicle diameter was 16 mm (range 11-23 mm). Follicle fluid VEGF concentrations (mean 6900 pg/ml, range 1200-17 100 pg/ml) were correlated positively with follicle fluid progesterone concentrations (mean 10 176 nmol/l, range 636-66780 nmol/l, r=0.62, P=0.002). This correlation was even tighter (r=0.87, P < 0.0001) when only samples from the 22 women in the earliest stages of follicle luteinization were considered. In these women serum LH concentrations were also correlated with follicle fluid VEGF concentrations (r=0.51, P=0.02). Our findings demonstrate the close dynamic relationship between VEGF production and early luteinization in human follicles during normal non-stimulated cycles.     

Microsurgical epididymal sperm aspiration with motile trophozoite cells but no spermatozoa

This paper reports on a patient in whom the clinical diagnosis of obstructive azoospermia was made according to clinical observations, i.e. azoospermia, normal andrological examination, normal follicle stimulating hormone and a misleading histopathological report of a testicular biopsy. Microsurgical vasoepididymostomy failed to restore fertility, and as a last resort, microsurgical sperm aspiration was performed. Although flagellated cells were observed in the epididymal aspiration, no spermatozoa were observed and wet preparation of multiple testicular biopsies failed to demonstrate any spermatozoon. This patient was diagnosed to have a non-obstructive azoospermia, resulting from maturation arrest associated with trichomonas infection at the level of the epididymis.     

Expression of ribonucleolytic toxin restrictocin in Escherichia coli: purification and characterization

OBJECTIVES: To define the incidence and severity of ovarian hyperstimulation syndrome (OHSS) occurring in oocyte donors. METHODS: Women (n = 149) aged 31.3 +/- 4.8 years (mean +/- S.D., range 21-41 years) participated as designated oocyte donors and underwent 400 consecutive cycles of controlled ovarian stimulation using human menopausal gonadotropin following pituitary downregulation with gonadotropin-releasing agonist. Patients were monitored by serial transvaginal ultrasound examinations and serum estradiol (E2) determinations. Oocytes (15.6 +/- 7.5 per aspiration; range 2-57) were harvested by ultrasound-directed transvaginal follicle aspiration 36 h following the intramuscular injection of human chorionic gonadotropin (hCG). Follow-up examination occurred 1 and 2 weeks post-aspiration. RESULTS: On the day of hCG injection E2 levels ranged from 512 to 13,502 pg/ml (mean 2902.7 +/- 1486.9 pg/ml). Over the next few weeks the degree of hyperstimulation in donors was staged: mild 65% (grade I, n = 98; grade II, n = 162); moderate 33.5% (grade III, n = 120; grade IV, n = 14); severe 1.5% (grade V, n = 6; grade VI, n = 0). Associated preaspiration E2 levels were: grade I, 1120 +/- 424 pg/ml; grade II, 2084 +/- 613 pg/ml; grade III, 3785 +/- 1713 pg/ml; grade IV, 5370 +/- 1264 pg/ml; grade V, 4286 +/- 1100 pg/ml. Worsening OHSS was associated with increasing levels of E2. There were no serious complications and hospitalization was not required. All symptoms resolved within 30 days of aspiration, disappearing by the time of the first menstrual flow in women of grade-III or lower stage. CONCLUSION: Although oocyte donors commonly experienced exaggerated levels of serum E2 they rarely (< 2%) developed severe OHSS. This may be attributable to their lack of embryo transfer which avoids exacerbating the illness.     

Ovarian abscess after ovum retrieval for in-vitro fertilization

An ovarian abscess is an uncommon surgical emergency that could be lethal. The causes of an ovarian abscess vary, and treatment thereof may unfortunately lead to an oophorectomy. In order to draw the attention of physicians to this rare entity, we present a case of ovarian abscess resulting from follicle aspiration for in-vitro fertilization. Furthermore, with correct preoperative diagnosis and prompt surgical intervention at an early stage, the affected ovary may be salvageable.     

Characterization of LeMir, a root-knot nematode-induced gene in tomato with an encoded product secreted from the root

The role of endoscopic ultrasound (EUS) in the detection and staging of pancreatic cancer is well established in medical literature. The development of EUS-guided fine needle aspiration (FNA) and subsequently EUS-guided fine needle injection (FNI) has expanded the clinical utility of EUS. These newer techniques made "interventional" EUS possible. Several recent applications of EUS-guided FNI include celiac nerve block, pseudocyst drainage, and drug delivery into pancreatic tumors. EUS is also gaining acceptance as an alternative diagnostic modality in the management of choledocholithiasis. The value of EUS in the diagnosis of early chronic pancreatitis is still being actively studied. This article reviews a number of recent developments in EUS-guided diagnosis and therapy with an emphasis on EUS-guided FNA and EUS-guided FNI.     

Are dysfunctional attitudes in depressive disorder trait or state dependent?

BACKGROUND: Fifty-one cases of de novo fibroadenoma in women aged 35 years and older were found during an analysis of 117,729 visits to the Wesley Breast Clinic from 1990 to 1996. METHODS: The clinical, mammographic and ultrasound diagnosis of fibroadenoma was confirmed by either fine needle aspiration cytology or histology of an open biopsy specimen. In all cases there was a well-documented previous visit available for review, at which there was no clinical or radiological evidence of the fibroadenoma. RESULTS: Thirty-seven of the de novo fibroadenomas were palpable, the remainder satisfying strict mammographic and/or ultrasound criteria. Four of the new fibroadenomas were in women aged 50-52. CONCLUSIONS: This study provides information about the natural history of fibroadenomas, confirming that they can appear for the first time in middle-aged women. This has important clinical implications, since new lesions appearing in women over 35 have tended to be automatically categorized as suspicious of carcinoma. However, a multidisciplinary approach involving clinical examination, mammography, ultrasound, and fine needle aspiration cytology or core biopsy can result in a confident diagnosis of fibroadenoma. This will allow some women with new lesions to be managed conservatively rather than by open biopsy.     

Role of diameter differences among follicles in selection of a future dominant follicle in mares

Follicles > or = 5 mm were ablated in pony mares by a transvaginal ultrasound-guided technique on Day 10 (ovulation = Day 0). Follicle emergence (at 15 mm, experiment 1; at 6 mm, experiment 2) and development of the new wave was monitored by transrectal ultrasound. Deviation was defined as the beginning of a marked difference in growth rates between the two largest follicles. In experiment 1, mares were grouped (n = 4 per group) into controls, ablation-controls (ablations at Day 10 only), and a two-follicle model (periodic ablation sessions so that only the two largest follicles developed). There were no significant indications that the two-follicle model altered follicle diameters, growth rates, or time intervals of the two retained follicles at or between events (follicle emergence, deviation, and ovulation). In experiment 2, the two-follicle model (n = 14) was used for follicle and hormonal characterization and hypothesis testing, without the tedious and error-prone necessity for tracking many (e.g., 20) individual follicles. The future dominant follicle emerged a mean of 1 day earlier (p < 0.008) than the future subordinate follicle, the growth rates for the two follicles between emergence and deviation (6 days later) did not differ, and the dominant follicle was larger at the beginning of deviation (23.1 +/- 0.8 mm versus 19.6 +/- 0.9 mm; p < 0.0001). Mean FSH and LH concentrations increased (p < 0.05) concomitantly from emergence of the future dominant follicle and peaked 3 days later when the follicle was a mean of 13 mm. Thereafter, the two hormones disassociated until ovulation: FSH decreased and LH increased. Results supported the hypothesis that the future dominant follicle has an early size advantage over future subordinate follicles and indicated that the advantage was present as early as 6 days before deviation.