A theoretical model for single blastocyst transfer.

OBJECTIVE: To elucidate the relationship between embryo grade and ART outcomes and determine how to decrease multiple pregnancy rates by assigning patients to single embryo transfer (SET) instead of dual embryo transfer (DET) according to embryo grade and/or availability. DESIGN: Retrospective medical record review. SETTING: IVF fertility center. PATIENTS: 247 women undergoing day 5 DET after in vitro fertilization or intracytoplasmic sperm injection treatment. METHODS: We retrospectively investigated embryo grade and outcomes on day 5 DET and calculated theoretical multiple pregnancy rates by assigning patients to SET instead of DET according to a combination of embryo grades and availability. MAIN OUTCOMES MEASURES: Implantation, pregnancy, multiple pregnancy, expected pregnancy and expected multiple pregnancy rates. RESULTS: Embryo grade affects implantation rates so that embryo transfer of at least one embryo with grade > or = 3BB resulted in high multiple pregnancy rates as well as high pregnancy rates. By performing DET, the clinical pregnancy rate was 39.9% with a multiple pregnancy rate of 33.3%; however, had SET been performed with an embryo graded 3BB or better, theoretical calculated pregnancy rates would have dropped to 35.8% but with a multiple pregnancy rate of 7.2%. CONCLUSIONS: Our model showed that cases having at least one embryo with grade > or = 3BB might serve as suitable candidates for SET.

Absence of estrogen receptor-alpha expression in human ovarian clear cell adenocarcinoma compared with ovarian serous, endometrioid, and mucinous adenocarcinoma.

The mechanism that regulates growth in ovarian clear cell adenocarcinoma (CCA) is not well understood. A high incidence of concurrent endometriosis with CCA may indicate that estrogen is a growth promotor in CCA. To determine estrogen as a growth promotor, the authors investigated the presence or absence of estrogen receptor-alpha (ER-alpha), ER-beta, progesterone receptor, and dioxin receptor (i.e., aromatic hydrocarbon receptor) in clinically resected ovarian CCA, serous adenocarcinoma (SAC), endometrioid adenocarcinoma (EAC), and mucinous adenocarcinoma (MAC) specimens using an immunohistochemical method. Expression of ER-alpha and ER-beta messenger ribonucleic acid was examined by reverse transcription-polymerase chain reaction in three established CCA cell lines: KK, RMG-1, and HAC-II. None of the surgically resected CCA and CCA cell lines showed positive staining for ER-alpha. Conversely, 97.2% of SACs, 100% of EACs, and 70% of MACs showed positive nuclear staining for ER-alpha (p < 0.001). Conversely, positive ER-beta staining for CCA (39.3%) was similar to that of SAC (41.7%) and MAC (30.0%). EAC (75%) showed a higher expression of ER-beta (p < 0.02). Progesterone receptor was detected in only 10.7% of CCA, compared with SAC and EAC (SAC, 86.1%; EAC, 91.7%; p < 0.01). Aromatic hydrocarbon receptor was detected in all histologic types at an incidence of approximately 50% to 60%. Messenger ribonucleic acid of ER-alpha and ER-beta was not detected in the three CCA cell lines. These findings indicate biologic characteristics that distinguish CCA from other types of ovarian epithelial cancer.

The acoustic calls of blue whales off California with gender data.

The acoustic calls of blue whales off California are described with visual observations of behavior and with acoustic tracking. Acoustic call data with corresponding position tracks are analyzed for five calling blue whales during one 100-min time period. Three of the five animals produced type A-B calls while two produced another call type which we refer to as type D. One of the animals producing the A-B call type was identified as male. Pauses in call production corresponded to visually observed breathing intervals. There was no apparent coordination between the calling whales. The average call source level was calculated to be 186 dB re: 1 muPa at 1 m over the 10-110-Hz band for the type B calls. On two separate days, female blue whales were observed to be silent during respective monitoring periods of 20 min and 1 h.

A case of congenital mumps infection complicated with persistent pulmonary hypertension.

A low-birth-weight female baby was admitted with respiratory distress after birth. Her mother had been diagnosed with mumps 4 weeks and 5 days prior to delivery. Mumps IgM antibody was elevated in the neonate and mumps virus ribonucleic acid was detected in the umbilical cord blood by reverse transcription-polymerase chain reaction. The perinatal virus infection was complicated with persistent pulmonary hypertension of the newborn and pulmonary hemorrhage. Successful treatment included the use of high frequency oscillation ventilation together with the administration of artificial surfactant.

Anxiety-like behavior in transgenic mice with brain expression of neuropeptide Y.

Neuropeptide Y (NPY), one of the most abundant peptide transmitters in the mammalian brain, is assumed to play an important role in behavior and its disorders. To understand the long-term modulation of neuronal functions by NPY, we raised transgenic mice created with a novel central nervous system (CNS) neuron-specific expression vector of human Thy- gene fragment linked to mouse NPY cDNA. In situ hybridization analysis demonstrated transgene-derived NPY expression in neurons (e.g., in the hippocampus, cerebral cortex, and the arcuate nucleus of the hypothalamus) in the transgenic mice. The modest increase of NPY protein in the brain was demonstrated by semiquantitative immunohistochemical analysis and by radioreceptor assay (115% in transgenic mice compared to control littermates). Double-staining experiments indicated colocalization of the transgene-derived NPY message and NPY protein in the same neurons, such as in the arcuate nucleus. The transgenic mice displayed behavioral signs of anxiety and hypertrophy of adrenal zona fasciculata cells, but no change in food intake was observed. The anxiety-like behavior of transgenic mice was reversed, at least in part, by administration of corticotropin-releasing factor (CRF) antagonists, alpha-helical CRF9-41, into the third cerebral ventricle. These results suggest that NPY has a role in anxiety and behavioral responses to stress partly via the CRF neuronal system. This genetic model may provide a unique opportunity to study human anxiety and emotional disorders.

Neuropeptide Y produces anxiety via Y2-type receptors.

In the present study, the effects of intracerebroventricular (ICV) NPY, [Leu31, Pro34]NPY and NPY13-36 have been evaluated with respect to anxiety in mice in the elevated plus maze. NPY had opposing effects on behavior, depending on the doses used. NPY decreased the normal preference for the closed arms of the maze at 0.7 nmol, indicating an anxiolytic effect; however, at 7 pmol NPY further increased the preference for the closed arm, indicating an anxiogenic effect. [Leu31, Pro34]NPY, a Y1-type receptor agonist, significantly reduced the preference for the closed arms at 70 pmol. NPY13-36, a Y2-type receptor agonist, significantly intensified the preference at 20 pmol. It has been demonstrated that NPY produces not only an anxiolytic effect via Y1-type receptors, but also an anxiogenic effect via Y2-type receptors. The time course of these NPY actions are quite different and the anxiogenic effect was observed only shortly after ICV NPY injection.

Dynorphin binds to neuropeptide Y and peptide YY receptors in human neuroblastoma cell lines.

The modulation of neuropeptide Y (NPY) and peptide YY (PYY) receptors by dynorphin, luteinizing hormone-releasing hormone (LHRH), corticotropin-releasing factor (CRF), and cholecystokinin octapeptide has been studied in human neuroblastoma cell lines SK-N-MC and SMS-MSN, which express Y1 and Y2 receptors for NPY/PYY. Dynorphin A and LHRH inhibited the binding of NPY/PYY to SK-N-MC cell membranes at concentrations ranging from 10(-7) to 10(-5) M, whereas dynorphin A and CRF were effective in SMS-MSN cells. The inhibitory effect of dynorphin A on NPY/PYY binding was observed in the presence of guanosine 5'-O-(3-thiotriphosphate), a nonhydrolyzable GTP analogue, as well as H-7 and H-8, novel inhibitors of protein kinases C and A. However, U-50488, the most potent kappa-selective compound did not mimic the dynorphin action. Dynorphin A showed neither effect on the dissociation of NPY/PYY from their receptors nor inhibition on the basal as well as forskolin-stimulated adenosine 3',5'-cyclic monophosphate response. These results indicate that the interaction of dynorphin A with Y1 and Y2 receptors is not mediated by changes in receptor-G protein interaction, receptor phosphorylation, and allosteric binding to NPY/PYY receptors but that dynorphin A binds to NPY/PYY receptors at high concentrations, probably in an antagonistic manner.

Effects of pancreatic polypeptide family peptides on feeding and learning behavior in mice.

We studied the effects of intra-third cerebroventricular administration of neuropeptide Y (NPY), peptide YY (PYY) and pancreatic polypeptide (PP) on the locomotor activity and the feeding and learning behavior of mice. NPY (0.3-10 micrograms), PYY (0.1-10 micrograms) and PP (3.0-10 micrograms) produced significant increases in locomotor activity. A significant decrease was then observed 15 min after administration of 10 micrograms of PYY. NPY, PYY and PP significantly increased food intake at 20 min and this effect continued for 2 to 4 hr at the high doses. The feeding response to PP family peptides were quite similar to that in locomotor activity with respect to dose-response, time course and peptide specificity. Learning behaviors were evaluated at three different stages of memory processing, acquisition, consolidation and retrieval, in a battery of step-down type passive avoidance tests. NPY and PYY had no effect on acquisition, but significantly improved consolidation at a dose of 0.03 and 0.3 microgram, respectively. NPY also improved retrieval at a dose of 0.03 microgram. The ranking order of potency in stimulating feeding and locomotor activity was PYY > NPY > PP, and in improving memory consolidation NPY > PYY >> PP. These observations suggest that NPY and PYY influence different neural substrates in the brain involved in feeding and learning.

Cholecystokinin octapeptide analogues suppress food intake via central CCK-A receptors in mice.

To examine the mechanism of the satiety-producing effect of cholecystokinin (CCK) in the central nervous system, we compared the potency of intraperitoneally (ip) or intracerebroventricularly (icv) administered CCK-8 and its analogues on food intake in fasted mice. The icv administration of a small dose of CCK-8 (0.03 nmol/brain) or of Suc-(Thr28, Leu29, MePhe33)-CCK-7 (0.001 nmol/brain) suppressed food intake for 20 min, whereas CCK-8 (1 nmol/kg, which is equivalent to 0.03 nmol/brain) or Suc-(Thr28, Leu29, MePhe33)-CCK-7 (1 nmol/kg) had satiety effect after ip administration. Dose-response studies indicated the following rank order of potency: Suc-CCK-7 > or = Suc-(Thr28, Leu29, MePhe33)-CCK-7 > or = CCK-8 > or = (Nle28,31)-CCK-8 >> desulfated CCK-8 = CCK-4 = 0 in the case of ip administration and Suc-(Thr28, Leu29, MePhe33)-CCK-7 >> Suc-CCK-7 > or = CCK-8 > or = (Nle28,31)-CCK-8 >> desulfated CCK-8 = CCK-4 = 0 in the case of icv administration. The selective CCK-A receptor antagonist MK-329 reversed the inhibitory effect of the centrally as well as peripherally administered CCK-8, or of Suc-(Thr28, Leu29, MePhe33)-CCK-7, whereas the selective CCK-B receptor antagonist L-365260 did not. The icv administered CCK-8 did not appear in the peripheral circulation. These findings suggest the participation of CCK-A receptors in the brain in mediating the satiety effect of CCK and the difference in CCK-A receptors in the brain and peripheral tissues.

Structural requirements for the effects of neuropeptide Y on the hypothalamic-pituitary-adrenal axis in the dog.

Neuropeptide Y (NPY), administered into the third cerebral ventricle of the dog stimulates plasma ACTH and cortisol secretion. To further investigate the structure-activity relationships of this action, we examined the effect of COOH-terminal fragment, NPY 19-36 and its analogues, NPY-(1-36)-OH (deamidated NPY) and avian pancreatic polypeptide (APP) on ACTH-cortisol secretion following intracerebroventricular (i.c.v.) injection in the dog. NPY (1.19 nmol) evoked a significant increase in the secretion of both plasma ACTH and cortisol. However, NPY 19-36 and NPY-(1-36)-OH each failed to increase plasma ACTH and cortisol secretion at doses of 1.19-11.9 nmol injected i.c.v. APP was less active than NPY. These data demonstrate that the entire NPY molecule is required for the full expression of the stimulatory effect of NPY on the secretion of ACTH and cortisol.

Solubilization of the receptors for avian pancreatic polypeptide in chicken, canine, and pig brains.

When n-octyl-beta-D-glucoside was used in several detergents to extract active avian pancreatic polypeptide (APP) receptors, a specific binding of [125I]APP to the solubilized chicken cerebellar and porcine hippocampal membranes was found. The binding of [125I]APP to the solubilized receptors was dependent on incubation time, temperature, and protein concentrations and appeared to have a slightly acidic optimal pH. APP binding to chicken and porcine brain extracts showed a high specificity for APP, although the chicken receptors do not discriminate well between APP and its related peptides, neuropeptide Y and peptide YY. Scatchard analyses of competitive binding data indicated the presence of two classes of binding sites in the brain extracts as in membrane-bound receptors; however, the high affinity component of the chicken receptor showed a decreased affinity after extraction. APP receptors in chicken and porcine brain extracts retained their insensitivity to the nonhydrolyzable GTP analog guanosine 5'-O-(3-thiotriphosphate). Cross-linking studies were performed with the homobifunctional cross-linker disuccinimidyl suberate and brain membrane receptors solubilized with n-octyl-beta-D-glucoside. An APP receptor species with a M(r) of 67,000, the same size as that of the labeled protein in native membrane homogenates of chicken and pig brains, was identified. However, in the canine brain we observed a M(r) 85,000 receptor protein, suggesting that species differences exist among the structures of brain APP receptors. The solubilized cross-linked APP receptors in these species were adsorbed by wheat germ agglutinin-agarose and by concanavalin A, indicating that they are glycoprotein in nature. The availability of the solubilized receptors from vertebrate brains with n-octyl-beta-D-glucoside represents an important step toward the purification and molecular characterization of the APP receptors.