HDLive ultrasound images of ovarian dermoid cysts: diagnostic accuracy.

OBJECTIVE: To demonstrate that the use of 3D/4D HDLive increases the image quality in the diagnosis of benign cystic ovarian teratomas. MATERIALS AND METHODS: 3D/HDLive ultrasound (US) was used in 31 cases of suspected ovarian cystic teratoma using vaginal 2D US. The following pathognomonic images of mature cystic teratomas were considered for diagnosis: 1) a cystic, unilocular lesion with a densely echogenic tubercle (Rokitansky nodule); 2) a diffuse or partially echogenic mass usually demonstrating sound attenuation; 3) fluid-fluid/fat-fluid levels; 4) dermoid mesh with hyperechogenic calcifications indicating the presence of bone, teeth, or other ectodermally-derived structure; 5) multiple mobile spherical structures (fat globules). RESULTS: Dermoids present a wide spectrum of images depending on the predominant tissue type. In the vast majority of cases there are dense echogenic structures that correspond to complex masses of fatty tissue, sebum, hair, epithelial remnants, along with cartilage or bone. If we catalogue all the images together, the pathognomonic of dermoid are: 1) cystic or solid cystic lesions with a Rokitansky nodule, with bone, teeth or cartilage (six cases, 22.2%); 2) a solid mass with or without attenuation that corresponds with pure sebum (five cases, 18.5%); 3) a diffuse mass with fine bands that correspond with hair inside sebum (four cases, 12.9%) and that may form meshes or plugs corresponding with a mixture of fat, sebum, and hair (three cases, 11.5%). CONCLUSIONS: HDLive U.S. provides some images of exceptional quality that enhance the definition of the structures of these tumors (fat, hair, cartilage, bone, etc.) compared to 2D/3D/4D.

Semi-automatic Sono T measurement of nuchal translucency.

A prospective study of 63 singleton pregnancies between 11 + 0 and 13 + 6 weeks gestation underwent semi-automatic nuchal translucency (NT) measurement and were compared with two-dimensional ultrasonography (2D US). Inter-observer variation and the repeatability were evaluated. Sono T automatically achieves mid-sagittal plane views and measures the maximum NT thickness. Measurements have less inter-observer variation (CI = -0.13, -0.04) when compared with 2D measurements (CI = -0.45, 0.28). It is reproducible and comparable to conventional 2D US technique for NT measurement. However, incorporating Sono T into routine practice requires further program refinements in order to reduce erroneous NT measurements.

High doses of GnRH antagonists are efficient in the management of severe ovarian hyperstimulation syndrome.

OBJECTIVE: To determine whether treatment of severe ovarian hyperstimulation syndrome (OHSS) with high-dose gonadotropin-releasing hormone (GnRH) antagonist, due to its luteolytic effect, is an effective method of management. METHODS: Six infertile patients who had been scheduled for embryo transfer and developed early-onset severe OHSS with ascites and hemoconcentration were chosen for treatment with 3.0 mg of a GnRH antagonist (Cetrotide; Cetrorelix, Serono, Madrid, Spain). The response of these patients was compared with five patients with severe early-onset OHSS who received support therapy alone. All patients were evaluated clinically, echographically, and hematologically. RESULTS: Estradiol (E2) levels dropped significantly a few days after treatment. Peritoneal fluid regression measured by ultrasound was faster on the study group compared with controls. Hematocrit remained comparable in both groups during follow-up. In two cases a second bolus of GnRH-antagonist was used due to clinical and biochemical findings during the four days of observation following the initial dose. None of the patients treated with GnRH antagonists required paracentesis. CONCLUSIONS: Treatment with high doses of GnRH antagonists seems to be effective in the management of severe OHSS.

The role of gonadotropin-releasing hormone in murine preimplantation embryonic development.

Previous studies have established the presence of an extrahypothalamic GnRH in a variety of tissues. GnRH receptor is known to be present in the placenta, which produces and secretes the decapeptide from the very early stages of placentation. We hypothesized that GnRH may play a role in the preimplantation development of embryos. To examine this hypothesis, we assessed GnRH and GnRH receptor messenger RNA (mRNA; RT-PCR) and protein expression (Immunohistochemistry) in preimplantation murine embryos at various developmental stages. Furthermore, preimplantation murine embryos were cultured with GnRH agonist and antagonist in vitro to assess the influence of GnRH analogs on embryo development. GnRH is expressed in the developing mouse embryo from morula to hatching blastocyst stages at the mRNA and protein levels. GnRH receptor mRNA is also present in the developing embryos studied. Preimplantation embryonic development was significantly enhanced by incubation with increasing concentrations of GnRH agonist and is significantly decreased by GnRH antagonist compared with that in the control group. Moreover, GnRH antagonist (5 and 10 microM) was able to completely block embryo development. The deleterious effect of GnRH antagonist on embryo development was reversed by increasing concentrations of the agonist, as determined by the number of embryos reaching the blastocyst stage.

Human oviductal gonadotropin-releasing hormone: possible implications in fertilization, early embryonic development, and implantation.

The oviduct is host to gametes and early embryos at a critical point in their lives. It is clear that the interactions of gametes/early embryo with the maternal oviduct in an autocrine and paracrine manner provide a microenvironment that enhances fertilization, early embryonic development, and implantation. Moreover, there is considerable evidence that an extrahypothalamic GnRH may play a substantial role as a molecular autocrine/paracrine regulator in these events. Gametes and preimplantation embryos express GnRH and GnRH receptor at both messenger ribonucleic acid (mRNA) and protein levels. However, whether GnRH is produced by the human oviduct has not yet been demonstrated. We used RT-PCR and immunohistochemical techniques to investigate GnRH mRNA and protein expression in human fallopian tubes throughout the menstrual cycle of premenopausal fertile patients. Our results, at both the mRNA and protein levels, revealed cycle-dependent production of an oviductal GnRH with expression during the luteal phase. Moreover, GnRH immunostaining was localized in the tubal epithelium during the luteal phase. On the basis of these data, we suggest that during reproductive life, oviductal GnRH may play a substantial paracrine/autocrine role in human fertilization, early embryonic development, and implantation.

Interleukin (IL)-1beta regulation of IL-1beta and IL-1 receptor antagonist expression in cultured human endometrial stromal cells.

The interleukin (IL)-1 system is a major regulator of local cellular interactions during embryonic implantation. Because IL-1beta and IL receptor antagonist (IL-1ra) are both expressed in human endometrium, we hypothesized that an appropriate ratio of IL-1beta to IL-1ra might favor the process of embryo implantation. Therefore, we investigated IL-1 regulation of the quantitative ratio of IL-1beta/IL-1ra messenger RNA (mRNA) expression in human endometrial stromal cells using quantitative competitive PCR, as well as intracellular protein expression after stromal cell solubilization. Confluent stromal cell cultures were stimulated with human IL-1beta (0-1000 IU/mL) for 24 h. After 24 h, total RNA was extracted, reverse transcribed, and coamplified by PCR with a defined amount of internal standard. The quantitative ratio was determined by the density of target to the internal standard. After culture with IL-1beta for 24 and 48 h, stromal cells were solubilized, and the intracellular protein levels of IL-1beta and IL-1ra were measured by enzyme-linked immunosorbent assay. The IL-1beta and IL-1ra mRNA were both up-regulated, and IL-1R tI mRNA was down-regulated, by IL-1beta in a dose-dependent manner. The quantitative ratio of IL-1beta to IL-1ra mRNA was constant with the presence of increasing concentrations of IL-1beta (1-1000 IU/mL). IL-1beta and IL-1ra protein was not detected in conditioned media of cultures before addition of IL-1beta. IL-1beta and IL-1ra protein levels increased with increasing amounts of IL-1beta after solubilization of stromal cells. The IL-1beta was detectable after 12 h of culture, in comparison with IL-1ra, which was detectable after 24 h of IL-1beta stimulation. These results suggest that IL-1 may play a crucial role in embryo-maternal interaction by regulating stromal cell expression of IL-1beta and IL-1ra, resulting in an appropriate ratio during the process of embryonic implantation.

Independent regulation of matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1 (TIMP-1), and TIMP-3 in human endometrial stromal cells by gonadotropin-releasing hormone: implications in early human implantation.

Early human trophoblast shows dramatic invasive properties during early pregnancy. The simultaneous synthesis of matrix metalloproteinases (MMPs) and their specific tissue inhibitors (TIMPs) in both human trophoblast and decidual membranes suggests that their controlled and balanced expression is crucial for the rapid matrix remodeling and controlled invasion during early pregnancy. Recently, we have described the presence of an extrahypothalamic GnRH immunologically, biologically and chemically identical to the hypothalamic hormone in periimplantation human embryos. Moreover, the production of this decapeptide by the human trophoblast during the early stages of placentation is well documented. TIMP-1 and -3 messenger ribonucleic acid (mRNA) expression in cultured stromal cells and protein secretion into the medium were significantly decreased by GnRH agonist compared to that in control groups. Moreover, expression of TIMP-1 was affected to a greater extent than that of TIMP-3. GnRH antagonist ablated the down-regulation of TIMPs by the GnRH agonist. MMP-9 mRNA expression was not detected in the control groups or in the groups treated with GnRH analogs. Our results provide evidence that trophoblastic GnRH may play an important role in placental tissue organization and in the early embryo-maternal dialogue by enhancing trophoblast invasion through the specific inhibition of TIMPs.

Congenital Müllerian anomalies: diagnostic accuracy of three-dimensional ultrasound.

OBJECTIVE: To determine whether it is possible to identify and diagnose accurately Müllerian anomalies with three-dimensional (3-D) ultrasound (US). DESIGN: Controlled blinded clinical study. SETTING: Normal human volunteers undergoing infertility evaluation in a university hospital. PATIENTS: Forty-two patients who underwent laparoscopy and hysterosalpingography as part of their work up for infertility and were found to have either a normal uterus (30 patients) or a Müllerian abnormality (12 patients) consented to be evaluated with 3-D US by sonographers who were unaware of their infertility history or of their laparoscopy and hysterosalpingography diagnoses. INTERVENTIONS: Transvaginal 3-D US evaluation over a 10- to 15-minute duration. MAIN OUTCOME MEASURES: Three-dimensional imaging was successful in all cases. RESULTS: Sonographers identified a Müllerian anomaly in all cases and came up with the correct classification in 11 of 12 cases. All patients with a normal uterus were identified correctly. CONCLUSIONS: In all patients with Müllerian anomalies, 3-D US examination of the endometrial cavity correlated with hysterosalpingography. In 91.6% of patients, 3-D US correlated with the external uterine configuration observed by laparoscopy. This technique may be used reliably in an office setting to diagnose and classify Müllerian anomalies.

Immunoreactive gonadotropin-releasing hormone expression in cycling human endometrium of fertile patients.

OBJECTIVE: To investigate the protein expression of GnRH in the endometrium of fertile patients throughout the menstrual cycle. DESIGN: Prospective longitudinal study. SETTING: Department of Gynecology and Obstetrics, Reproductive Immunology Laboratory, Stanford University Medical Center. PATIENT(S): Twenty-two fertile premenopausal women submitted to laparoscopic surgery for benign gynecologic indications. None of the 22 women had endometriosis or pelvic inflammatory disease. INTERVENTION(S): An endometrial biopsy specimen using the Novak curette was obtained at the time of surgery. MAIN OUTCOME MEASURE(S): Protein expression and localization from unfractioned endometrial tissue was analyzed by immunohistochemistry. RESULT(S): Gonadotropin-releasing hormone is expressed at the protein level in both the endometrial stroma and epithelium throughout the entire menstrual cycle of fertile women. Immunostaining in the human epithelium reached maximal levels in the midluteal phase and was elevated in the stroma throughout the entire luteal phase. CONCLUSION(S): Our results demonstrate the presence of GnRH in the human endometrium at the protein level throughout the entire menstrual cycle of fertile women, with an increase in the luteal phase compared with the preovulatory endometrium.

Sex specific proteins and proteases present in the midguts of Anopheles albimanus (Diptera: Culicidae).

Proteins and proteases present in midgut tissues of sugar-fed Anopheles albimanus (Wiedemann) males and females were studied by 2-dimensional electrophoresis and zymograms using gelatin and hemoglobin as substrates. Protein patterns differed between sexes. Some proteins were similar in both sexes, but differed in intensity. Sex specific proteins and midgut proteases also were detected. These findings indicate the possibility of sex dependent regulation of midgut proteins and protease production.

Role of color flow Doppler ultrasonography in diagnosing velamentous insertion of the umbilical cord and vasa previa. A report of two cases.

BACKGROUND: Prenatal diagnosis of velamentous insertion of the umbilical cord is of practical importance since unsupported vessels in the amnion may be lacerated at the time of membrane rupture. Failure to diagnose ruptured vessels may lead to fetal death. The incidence of velamentous insertion of the umbilical cord is about 1% in singleton pregnancies. This mode of insertion occurs much more frequently with multifetal pregnancies. CASES: Two cases of velamentous insertion of the umbilical cord unrelated to vasa previa were diagnosed prenatally with color Doppler ultrasound in women with twin gestations. Both women delivered uneventfully by cesarean section. The velamentous insertion diagnosis was confirmed postnatally in both cases. CONCLUSION: Color flow Doppler ultrasonography can be used in the second and third trimesters of pregnancy to identify potentially lethal structural abnormalities of the umbilical cord.

Color flow Doppler: a useful instrument in the diagnosis of funic presentation.

Color Doppler sonography offers the opportunity to evaluate the umbilical cord and to study blood velocity wave forms within the cord. Funic (cord) presentation can be diagnosed unequivocally with color Doppler sonography. Because funic presentation is likely to be the harbinger of cord prolapse, the morbidity and mortality associated with cord prolapse can be prevented if funic presentation is diagnosed before membrane rupture. This article describes the first case in the obstetric literature of funic presentation diagnosed with color Doppler sonography. The potential complications associated with cord prolapse were avoided with a cesarean section.

Subchorionic placental cyst: a cause of fetal growth retardation--ultrasound and color-flow Doppler diagnosis and follow-up.

Subchorionic placental cysts are ominous findings. When attached near the umbilical cord insertion, the risk of umbilical cord constriction is increased. This may cause fetal growth retardation and intrauterine asphyxia. This article reports a case of subchorionic placental cyst diagnosed in the first trimester by transvaginal ultrasound. Color Doppler ultrasound demonstrated a reduction of the umbilical cord bloodstream as the cyst increased in size. Fetal growth retardation was evident in the third trimester.

Early diagnosis of conjoined twins using two-dimensional color Doppler and three-dimensional ultrasound.

Transvaginal three-dimensional (3-D) and color Doppler ultrasound were used to establish a first-trimester definitive diagnosis and classification of thoracoomphalopagus conjoined twins following two-dimensional (2-D) transabdominal and transvaginal scans that indicated twin gestation of uncertain classification. Color Doppler in combination with 3-D ultrasound can be a useful complement to 2-D ultrasound to confirm early diagnosis and determine the extent of organ sharing and definitive classification of conjoined twins.

Thanatophoric dysplasia: ultrasound diagnosis.

Six pregnant women with singleton fetuses were referred to our Prenatal Diagnostic Centers because of an initial diagnosis of intrauterine growth restriction (n = 4) or short femoral length (n = 2). We first examined the patients with two-dimensional (2D) ultrasonography (US), and then with three-dimensional (3D) US, and arrived at a definitive diagnosis of thanatophoric dysplasia. Although in experienced hands a diagnosis of thanatophoric dysplasia can be made with 2D US before the 24th week of gestation, our experience suggests that 3D US is a valuable complement to 2D US for the evaluation of fetuses with skeletal dysplasias.

Morphological and proteomic characterization of midgut of the malaria vector Anopheles albimanus at early time after a blood feeding.

The midgut of anopheline mosquito is the entry of Plasmodium, the causative agent of malaria.When the mosquito feeds on parasite infected host, Plasmodium parasites reach the midgut and must confront digestive enzymes, the innate immune response and go across the peritrophic matrix (PM), a thick extracellular sheath secreted by the mosquito midgut epithelial cells. Then, to continue its development, the parasite must reach the salivary glands to achieve transmission to a vertebrate host. We report here the morphological and biochemical descriptions of the midgut changes after a blood meal in Anopheles albimanus. Before blood feeding, midgut epithelial cells contained numerous electrondense vesicles distributed in the central to apical side. These vesicles were secreted to the luminal side of the midgut after a blood meal. At early times after blood ingest, the PM is formed near microvilli as a granulous amorphous material and after it consolidates forming a highly organized fibrillar structure, constituted by layers of electrondense and electronlucent regions. Proteomic comparative analysis of sugar and blood fed midguts showed several molecules that modify their abundance after blood intake; these include innate immunity, cytoskeletal, stress response, signaling, and digestive, detoxifying and metabolism enzymes. Biological significance In the midgut of mosquitoes during bloodfeeding, many simultaneous processes occur, including digestion, innate immune activities, cytoskeleton modifications, construction of a peritrophic matrix and hormone production, between others. Mechanical forces are very intense during bloodfeeding and epithelial and muscular cells must resist the stress, modifying the actin cytoskeleton and coordinating intracellular responses by signaling. Microorganisms present in midgut contents reproduce and interact with epithelial cells triggering innate immune response. When infectious agents are present in the blood meal they must traverse the peritrophic matrix, an envelope formed from secretion products of epithelial cells, and evade the immune system in order to reach the epithelium and continue their journey towards salivary glands, in preparation for the transmission to the new hosts. During all these processes, proteins of mosquitoes are modified in order to deal with mechanical and biological challenges, and the aim of this work is to study these changes.