Interventions for 'poor responders' to controlled ovarian hyperstimulation (COH) in in-vitro fertilisation (IVF).

BACKGROUND: The success of in-vitro fertilisation (IVF) treatment depends on adequate follicle recruitment by using controlled ovarian stimulation with gonadotrophins. Failure to recruit adequate follicles is called 'poor response'. Various treatment protocols have been proposed that are targeted at this cohort of women, aiming to increase their ovarian response. OBJECTIVES: To compare the effectiveness of different treatment interventions in women who have poor response to controlled ovarian hyperstimulation (are poor responders) in the context of in vitro fertilisation. SEARCH STRATEGY: We searched the Cochrane Menstrual Disorders and Subfertility Group Specialised Register of controlled trials (MDSG), the Cochrane Central Register of Controlled trials (CENTRAL) (The Cochrane Library 2003, Issue 1), MEDLINE (1966 to August 2006), EMBASE (1980 to August 2006) and The National Research Register (NRR). The citation lists of relevant publications, review articles, abstracts of scientific meetings and included studies were also searched. The authors were contacted to identify or clarify data that were unclear from the trial reports. SELECTION CRITERIA: Only randomised controlled trials (RCTs) comparing one type of intervention versus another for controlled ovarian stimulation of poor responders to a previous IVF treatment, using a standard long protocol were included. DATA COLLECTION AND ANALYSIS: Two review authors independently scanned the abstracts, identified relevant papers, assessed inclusion and trial quality and extracted relevant data. Validity was assessed in terms of method of randomisation, completeness of treatment cycle and co-intervention. Where possible, data were pooled for analysis. MAIN RESULTS: Nine trials involving six different comparison groups have been included in this review. Only one trial reported live birth rates. Four groups compared the long protocol with another intervention. Only one comparison group (bromocryptine versus long protocol) reported a higher clinical pregnancy rate per cycle, in the bromocryptine arm (OR 5.60, 95% CI 1.40 to 22.47). Two comparison groups showed a lower number of oocytes in the long protocol group (versus stop and gonadotrophin releasing hormone (GnRH) antagonist protocols). However, two comparison groups also showed lower cancellation rates in the long protocol treatment group (versus stop and GnRHa flare-up protocols). None reported any evident difference in the adverse effects. AUTHORS' CONCLUSIONS: There is insufficient evidence to support the routine use of any particular intervention either for pituitary downregulation, ovarian stimulation or adjuvant therapy in the management of poor responders to controlled ovarian stimulation in IVF. More robust data from good quality RCTs with relevant outcomes are needed.

Randomized controlled trial comparing couple satisfaction with appointment and telephone follow-up consultation after unsuccessful IVF/ICSI treatment.

Couples undergoing in vitro fertilization and intracytoplasmic sperm injection (IVF/ICSI) at Aberdeen Maternity Hospital come from a wide geographical area. Increasingly, telephone discussions after unsuccessful treatment have replaced appointments for those who do not live locally. The aim of this study was to compare patient satisfaction with telephone follow-up discussions versus clinic appointments. Couples were separated into those undergoing their first treatment cycle (100 couples) and those undergoing their second or subsequent treatment cycle (85 couples), and then randomized to either a telephone or appointment follow-up. Satisfaction was assessed by a postal questionnaire and analysis conducted on an 'intention to treat' basis. An overall response rate of 91% was achieved. Analysis indicated no statistically significant difference between telephone and appointment groups with regard to the degree of satisfaction. However, there was an association between the type of follow-up and the duration of discussion (P < 0.001): telephone follow-up discussions were significantly shorter than appointment follow-ups. There is the potential for significant savings in costs, both to the service and to patients, by providing telephone follow-up consultations. The savings may be achieved without compromising patient satisfaction as long as clinic appointments remain available as an option for those couples who prefer them.

Complement-induced phospholipase A2 activation in experimental membranous nephropathy.

BACKGROUND: In the passive Heymann nephritis (PHN) model of membranous nephropathy, C5b-9 induces glomerular epithelial cell (GEC) injury and proteinuria, which is partially mediated by eicosanoids. By analogy, in cultured rat GEC, sublytic C5b-9 injures plasma membranes and releases arachidonic acid (AA) and eicosanoids, due to activation of phospholipase A2 (PLA2). This study addresses the mechanisms of PLA2 activation. METHODS: PLA2 expression was assessed with the polymerase chain reaction or immunoblotting, and activity was determined using an in vitro assay or by measurement of free AA. RESULTS: Under basal conditions, GEC in culture expressed a relatively low level of cytosolic PLA2 (cPLA2) protein, while mRNAs of groups IB, IIA and V secretory PLA2s (sPLA2) were not detectable. Incubation of GEC with sublytic C5b-9 induced 1.5- to 2.0-fold increases in free [3H]AA at 40 minutes, and three and 24 hours. C5b-9 did not increase cPLA2 protein, and did not induce group IB, IIA or V sPLA2 mRNAs. Stable overexpression of cPLA2 in GEC amplified the C5b-9-induced increases in free [3H]AA, while analogous overexpression of group IIA sPLA2 had no effect. PLA2 activity was increased in glomeruli of rats with PHN, and this enhanced activity was characterized as cPLA2. There were no differences in cPLA2 protein expression between PHN and control glomeruli. CONCLUSIONS: Release of AA by C5b-9 in GEC in culture and in vivo is mediated by cPLA2, and the mechanism is consistent with post-translational regulation of cPLA2 activity. C5b-9 does not induce expression or stimulate activity of sPLA2 isoforms in GEC.

Complement C5b-9 induces receptor tyrosine kinase transactivation in glomerular epithelial cells.

In the passive Heymann nephritis (PHN) model of membranous nephropathy, C5b-9 induces glomerular epithelial cell (GEC) injury and proteinuria, which is partially mediated via production of eicosanoids. Using rat GEC in culture, we demonstrated that sublytic C5b-9 induced tyrosine phosphorylation of the epidermal growth factor receptor (EGF-R), Neu, fibroblast growth factor receptor-2, and hepatocyte growth factor receptor. In addition, C5b-9 stimulated increases in tyrosine(204) phosphorylation of extracellular signal-regulated kinase-2 (ERK2), as well as free [(3)H]arachidonic acid (AA) and prostaglandin E(2) (PGE(2)). Phosphorylated EGF-R bound the adaptor protein, Grb2, and the EGF-R-selective tyrphostin, AG1478, blocked the C5b-9-induced ERK2 phosphorylation, [(3)H]AA release, and PGE(2) production by 45 to 65%, supporting a functional role for EGF-R kinase in mediating the activation of these pathways. Glomeruli isolated from rats with PHN demonstrated increases in ERK2 tyrosine(204) phosphorylation and PGE(2) production, as compared with glomeruli from control rats, and these increases were partially inhibited with AG1478. Thus, C5b-9 induces transactivation of receptor tyrosine kinases, in association with ERK2 activation, AA release, and PGE(2) production in cultured GEC and glomerulonephritis in vivo. Transactivated tyrosine kinases may serve as scaffolds for assembly and/or activation of proteins, which then lead to activation of the ERK2 cascade and AA metabolism.

Role of extracellular matrix and Ras in regulation of glomerular epithelial cell proliferation.

Signals from extracellular matrix (ECM) to growth factor receptors regulate glomerular epithelial cell (GEC) proliferation. Epidermal growth factor (EGF), basic fibroblast growth factor, hepatocyte growth factor (HGF), or thrombin stimulated proliferation of GECs when the cells were adherent to collagen matrices, but not plastic substratum. Furthermore, EGF, HGF, or thrombin activated p42 mitogen-activated protein (MAP) kinase in collagen-adherent GECs, whereas activation was weak in GECs on plastic. To further examine the interaction of ECM with the Ras-MAP kinase cascade, GECs were stably transfected with a constitutively active Ras mutant (V12Ras). Low or moderate levels of V12Ras expression did not affect basal MAP kinase activity but, unlike parental GECs, in clones that express V12Ras, EGF was able to induce proliferation and activate MAP kinase when these cells were adherent to plastic. In parental and V12Ras-transfected GECs, MAP kinase activation was inhibited by cytochalasin D. Thus, adhesion of GECs to ECM facilitates proliferation and MAP kinase activation by mitogens acting via tyrosine kinase or non-tyrosine kinase receptors. Activation of pathway(s) downstream of V12Ras supplants signals from ECM that enable proliferation. These signals may involve the actin cytoskeleton.

Complement activates phospholipases and protein kinases in glomerular epithelial cells.

BACKGROUND: In rat membranous nephropathy, complement C5b-9 induces glomerular epithelial cell (GEC) injury and proteinuria, which in some models is partially mediated by eicosanoids. In cultured rat GEC, sublytic C5b-9 injures plasma membranes and releases arachidonic acid (AA) and eicosanoids, due to activation of cytosolic phospholipase A2 (cPLA2). In this study, we address the role of protein kinases in cPLA2 activation. METHODS: GEC were stably transfected with cDNAs of wild-type (wt) cPLA2, and serine505-->alanine mutant (cPLA2-SA505), which lacks the mitogen-activated protein kinase (MAPK) phosphorylation site. RESULTS: Complement stimulated protein kinase C (PKC) activity in GEC, and activated p42 (but not p38) MAPK. Overexpression of either cPLA2-wt or cPLA2-SA505 markedly amplified the release of [3H]AA by C5b-9. Depletion of PKC blocked the complement-dependent activation of cPLA2-wt or cPLA2-SA505, but inhibition of the p42 MAPK pathway had no effect. Epidermal growth factor was a strong activator of p42 MAPK, but stimulated PKC activity weakly. Unlike complement, activation of cPLA2-wt by epidermal growth factor was dependent on PKC, and was augmented significantly by p42 MAPK. Stable overexpression of phospholipase C-gamma 1 in GEC amplified C5b-9-induced production of [3H]inositol phosphates and [3H]diacylglycerol, an endogenous activator of PKC, and complement stimulated tyrosine phosphorylation of phospholipase C-gamma 1. CONCLUSIONS: C5b-9 induces activation of cPLA2 that is dependent on the diacylglycerol-PKC pathway. The role of p42 MAPK in cPLA2 activation becomes redundant in the presence of relatively potent PKC activation.

Activation of phospholipase A2 by complement C5b-9 in glomerular epithelial cells.

In rat membranous nephropathy, C5b-9 induces glomerular epithelial cell (GEC) injury and proteinuria, which is partially mediated by eicosanoids. In cultured rat GEC, sublytic C5b-9 injures plasma membranes and releases arachidonic acid (AA) and eicosanoids, due to activation of phospholipase A2 (PLA2). To address mechanisms of PLA2 activation, GEC were stably transfected with cDNAs of wild-type cytosolic PLA2 (cPLA2-wt), or group II secretory PLA2, producing overexpression of PLA2 activity. Sublytic C5b-9 markedly increased free [3H]AA in cPLA2-wt-transfected GEC, but only trivial increases were evident in secretory PLA2-transfected, or neo (control) GEC. In cPLA2-wt-transfected GEC, reduction of extracellular free Ca2+ or down-regulation of protein kinase C inhibited [3H]AA release. To further address the regulation of cPLA2, we stably expressed a mutant cPLA2 in which the Ca2+-dependent lipid binding domain was deleted (deltaCaLB). In GEC that express cPLA2-deltaCaLB, the C5b-9-induced increase in free [3H]AA was comparable with neo, despite expression of cPLA2-deltaCaLB at levels similar to cPLA2-wt. We then stably expressed another cPLA2 mutant (cPLA2-srcmyr) in which the CaLB domain was replaced by the N-terminal myristoylation domain of c-Src. cPLA2-srcmyr is permanently membrane associated. At low extracellular free Ca2+, C5b-9 increased free [3H]AA significantly in GEC that express cPLA2-srcmyr, while in neo GEC, the change was negligible. Thus, C5b-9 activates the cPLA2 isoform. Activation is dependent on the CaLB domain, and is mediated by phosphorylation, Ca2+ influx, and membrane association.

Receptor protein tyrosine kinases in perinatal developing rat kidney.

We have identified receptor protein tyrosine kinases (PTKs) that are expressed and/or activated during kidney development. mRNA from fetal rat kidneys in late gestation (embryonic day 21), was used to prepare a cDNA template for polymerase chain reaction amplification with primers based on conserved regions of PTKs, and products were subcloned and sequenced. Among 346 clones, we identified epidermal growth factor receptor (EGF-R), Tie-2, platelet-derived growth factor receptor (PDGF-R)-alpha, PDGF-R beta, Flk-1, Flt-4, fibroblast growth factor receptor (FGF-R)-1, FGF-R3, FGF-R4, Met, and RYK/Nbtk-1. PTK expression was studied by immunoprecipitation and immunoblotting of kidney membrane proteins with specific antibodies. EGF-R, PDGF-R alpha, FGF-R1, FGF-R3, Met, and in some cases Tie-2 protein expression was greater in fetal kidneys, as compared with kidneys from 12-week-old adult rats (controls). Flk-1, PDGF-R beta, and FGF-R4 proteins were expressed comparably, however, Flt-4 was not detected. As a reflection of receptor PTK activity, we assessed endogenous tyrosine phosphorylation, and in vitro autophosphorylation. EGF-R and PDGF-R alpha displayed activity in fetal, but not adult kidneys. FGF-R3 and Flk-1 were active in some fetal kidneys, and the other PTKs were not active. Thus, in late gestational rat kidney, there are distinct patterns of receptor PTK expression and activity. EGF-R, PDGF-R alpha, FGF-R3 and Flk-1 are among the PTKs that are activated, and they may mediate perinatal development of renal epithelial, interstitial, or vascular structures.

Extracellular matrix is required for MAP kinase activation and proliferation of rat glomerular epithelial cells.

This study examined the role of extracellular matrix (ECM) in the regulation of glomerular epithelial cell (GEC) proliferation. Epidermal growth factor (EGF) stimulated proliferation of GEC when the cells were adherent to collagen matrices, but not plastic substratum. Significant and prolonged EGF receptor (R) tyrosine autophosphorylation (which reflects EGF-R kinase activation) was induced by EGF only in GEC adherent to collagen. In addition, EGF stimulated the activity and tyrosine phosphorylation of p42 mitogen-activated protein (MAP) kinase (ERK2) in collagen-adherent GEC, but not in cells on plastic. An inhibitor of the p-42 MAP kinase pathway, PD98059, blocked EGF-induced MAP kinase activity and proliferation. Thus, adhesion to ECM enables EGF to induce proliferation of GEC, by facilitating activation of EGF-R and the p42 MAP kinase pathway. Signals from ECM to growth factor receptor tyrosine kinases may regulate cell turnover in the glomerulus under normal conditions and during immune glomerular injury.

Extracellular matrix stimulates production and breakdown of inositol phospholipids.

Adhesion of rat glomerular epithelial cells (GEC) to collagen stimulates production of D-myo-inositol 1,4,5-trisphosphate (IP3) and 1,2-diacylglycerol. This process is mediated via beta 1-integrins, and it modulates GEC proliferation. In this study, we address the changes in inositol-lipid turnover induced by GEC adhesion to extracellular matrix (ECM). The masses of both phosphatidylinositol 4,5-bisphosphate (PIP2) and IP3, as well as [3H]inositol phosphates, were increased in GEC adherent to collagen, compared with plastic substratum. Phosphatidylinositol-4-phosphate (PIP) 5-kinase activity was predominantly membrane associated and was enhanced in GEC on collagen. Phospholipase C (PLC) activity and PLC-gamma 1 protein were increased in membrane fractions of GEC adherent to collagen, compared with plastic. Stable overexpression of PLC-gamma 1 in GEC amplified the effect of ECM on the production of [3H]inositol phosphates. In addition, the PLC-gamma 1 that was membrane associated in collagen-adherent GEC was tyrosine phosphorylated. Thus production of IP3 in GEC adherent to ECM is associated with increased production of PIP2. Moreover, adhesion to ECM increases tyrosine phosphorylation and membrane association of PLC-gamma 1, which may facilitate PIP2 hydrolysis by increasing the catalytic activity of PLC-gamma 1 and the proximity of PLC-gamma 1 and its substrate. Understanding the process of ECM-induced inositol lipid production and breakdown in GEC may provide insights into the regulation of GEC proliferation and differentiated functions in normal conditions and during glomerular injury.

Complement C5b-9 activates cytosolic phospholipase A2 in glomerular epithelial cells.

In rat membranous nephropathy, complement C5b-9 induces glomerular epithelial cell (GEC) injury and proteinuria, which, in some models, is partially mediated by eicosanoids. By analogy, sublytic C5b-9 injures plasma membranes and releases arachidonic acid (AA) and eicosanoids in cultured rat GEC. In this study, we demonstrate that, in GEC, sublytic C5b-9 stably increased the activity of a high-molecular-mass cytosolic phospholipase A2 (PLA2), which we identified as "cPLA2." This increase was abolished with inhibitors of protein kinase C. C5b-9 did not affect low-molecular-mass membrane-associated or secretory PLA2 activities. In GEC that stably overexpress cPLA2 activity and protein (produced by transfection of cPLA2 cDNA), immunoblot analysis showed that sublytic C5b-9 induced a decreased mobility of cPLA2, consistent with cPLA2 phosphorylation. Incubation of cPLA2-transfected GEC with sublytic C5b-9 significantly increased production of free AA and prostaglandin E2, whereas, in control GEC, the C5b-9-induced changes in free AA and prostaglandin E2 were small. Furthermore, both C5b-9-dependent sublytic cytotoxicity and cytolysis were enhanced in GEC overexpressing cPLA2, compared with control cells. Thus C5b-9 increased cPLA2 activity, probably via phosphorylation involving a protein kinase C-dependent pathway. Phospholipid hydrolysis by cPLA2 resulted in release of substrate for eicosanoid synthesis and in enhancement of C5b-9-dependent GEC injury. Both processes may facilitate glomerular damage in membranous nephropathy.

Epidermal growth factor receptor activation in developing rat kidney.

Epidermal growth factor (EGF) binding increases in late-gestational rat kidney and then falls toward basal adult levels postnatally during the 1st wk. We report that the increase in EGF binding is accompanied by an increase in EGF receptor (EGFR) protein and activation of EGFR tyrosine kinase. Multiple proteins were endogenously tyrosine phosphorylated in kidney membranes from fetal rats, and the phosphorylation pattern was similar in rats ranging from 16 to 21 days of gestation. Tyrosine phosphorylation was, however, almost undetectable in 12-wk adult rat kidneys (controls). Among the phosphoproteins in fetal kidney, a prominent 170-kDa protein was identified as EGFR. Endogenous tyrosine phosphorylation of EGFR (reflecting receptor activation) was 30-fold higher in fetal kidney membranes than in adult (3- to 7-fold higher when adjusted for differences in EGF binding or EGFR protein content). The EGFR substrate, phospholipase C-gamma 1, was tyrosine phosphorylated in fetal kidneys but not adult, and a greater proportion was membrane-associated in fetal kidneys, consistent with activation of phospholipase C-gamma 1. Thus EGFR tyrosine kinase activity is increased in late-gestational rat kidney. Induction and activation of EGFR may mediate perinatal renal cell growth and development.

Extracellular matrix modulates epidermal growth factor receptor activation in rat glomerular epithelial cells.

To understand how glomerular epithelial cell (GEC) proliferation may be regulated in health and disease, we studied the effects of type I collagen extracellular matrices (ECM) on EGF receptor (EGF-R) activation in cultured rat GEC. EGF stimulated proliferation of GEC adherent to ECM, but not of GEC on a plastic substratum. Significant and prolonged EGF-R tyrosine autophosphorylation (which reflects receptor kinase activation) was induced by EGF in GEC adherent to collagen, but EGF did not stimulate EGF-R autophosphorylation in GEC on plastic (at 37 degrees C). However, EGF-R autophosphorylation increased significantly in plastic-adherent GEC that were stimulated with EGF at 4 degrees C or in the presence of vanadate, an inhibitor of phosphotyrosine phosphatases. Furthermore, dephosphorylation of EGF-R was enhanced in GEC on plastic as compared with collagen. At 4 degrees C, [125I]EGF binding was not different between substrata, and there was negligible accumulation of intracellular [125I]EGF (which reflects EGF-R internalization). At 37 degrees C, EGF-R internalization was reduced significantly in collagen-adherent GEC as compared with GEC on plastic. Thus, contact with ECM facilitates proliferation and EGF-R activation in GEC. The enhanced activity of EGF-R tyrosine kinase may be due to ECM-induced reduction in EGF-R internalization and dephosphorylation by phosphotyrosine phosphatase(s). Signals from ECM to growth factor receptors may regulate cell turnover in the glomerulus under normal conditions and during immune glomerular injury.

Extracellular matrix-stimulated phospholipase activation is mediated by beta 1-integrin.

beta 1-Integrins are major mediators of interactions between cells and extracellular matrix (ECM). Adhesion of rat glomerular epithelial cells (GEC) to collagen stimulated phospholipase C. As a result, 1,2-diacylglycerol (DAG) was increased, and inositol phospholipids were decreased in collagen-adherent cells, as compared with GEC adherent to plastic substrata. Adhesion to collagen also stimulated production of free arachidonic acid (the precursor for eicosanoids) due to metabolism of DAG through the DAG lipase pathway and due to phospholipase A2-induced hydrolysis of phospholipids. Phospholipase A2 appeared to be stimulated as a result of protein kinase C (PKC) activation, probably secondary to increased DAG. The collagen-induced increases in DAG and free arachidonic acid, as well as the decrease in inositol phospholipids, were partially inhibited by lowering extracellular Ca2+ concentration to 200 nM or less and by anti-beta 1-integrin antibody Fab. In contrast, anti-beta 1-integrin immunoglobulin G (IgG) enhanced collagen-mediated increases in DAG and arachidonic acid. Proliferation of GEC adherent to collagen was reduced in the presence of anti-beta 1-integrin IgG. The antiproliferative effect of anti-beta 1-IgG appeared to be mediated through PKC, since it was absent in PKC-depleted GEC. Immunoprecipitation with integrin subunit-specific antibodies demonstrated alpha 2 beta 1- and alpha 3 beta 1-integrins in GEC. Thus, in GEC, ECM induces activation of phospholipases C and A2, which is mediated, at least in part, by beta 1-integrins. Products of integrin-mediated phospholipase activation may modulate GEC proliferation.

Adhesion of rat glomerular epithelial cells to extracellular matrices: role of beta 1 integrins.

Glomerular epithelial cells (GEC) maintain glomerular permselectivity and are a target of immunological glomerular injury, which may lead to proliferation or detachment from extracellular matrix (ECM). We studied adhesion mechanisms in rat GEC in culture, focusing on adhesion molecules of the beta 1 integrin family. At early time points (1 hr after plating of cells into culture wells that had been pre-incubated with purified ECM proteins), adhesion of GEC to collagen I, collagen IV, laminin, and fibronectin was inhibited with anti-beta 1 integrin antibody. The peptide RGDS inhibited adhesion to fibronectin and laminin. Immunoprecipitation studies demonstrated the presence of alpha 2, alpha 3, and beta 1 integrins; the alpha 1, alpha 4, alpha 5, alpha 6, alpha v, and beta 3 subunits were undetectable. Adhesion to all ECM proteins was dependent on divalent cations, but the effects of individual cations varied among substrata. In rat GEC, alpha 2 beta 1 and/or alpha 3 beta 1 integrins appear to mediate adhesion to collagen I, collagen IV, and laminin. The alpha 3 beta 1 integrin is also the likely receptor for fibronectin, interacting through an RGD binding site. Furthermore, single integrins or combinations of integrins appear to have distinct ligand-binding functions that are differentially regulated by divalent cations. Characterization of GEC adhesion molecules may facilitate the understanding of mechanisms of glomerular development, and cell detachment or proliferation in immune glomerular injury.

Eicosanoids enhance epidermal growth factor receptor activation and proliferation in glomerular epithelial cells.

Proliferation of glomerular epithelial cells (GEC) and release of prostaglandins (PG) and thromboxane (Tx) A2 may occur in glomerular injury. We studied the relationship of eicosanoids to epidermal growth factor (EGF)-induced proliferation of rat GEC in culture. After 48 h of serum-deprivation, EGF stimulated [3H]thymidine incorporation ninefold above serum-deprived cells. Inhibition of cyclooxygenase with indomethacin or of Txsynthase with OKY-046 decreased the proliferative effect of EGF by 50 and 38%, respectively. The effect of indomethacin was reversed by addition of PGE2. Synthesis of PGE2, PGF2 alpha, and TxA2 by serum-deprived GEC was not enhanced by EGF. Scatchard analysis of 125I-EGF binding to GEC demonstrated two populations of EGF receptors; the high-affinity site had a dissociation constant (Kd) of 444 pM and 24,864 receptors/cell. EGF receptor autophosphorylation (reflecting receptor activation) was studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting of GEC membrane proteins with anti-phosphotyrosine antibody. EGF increased phosphorylation of a protein of approximately 170 kDa, which comigrated with proteins immunoprecipitated from [35S]methionine-labeled GEC with antibodies to EGF receptor. Indomethacin and OKY-046 decreased the EGF-dependent phosphorylation of the 170-kDa protein, and this decrease was overcome by addition of PGE2. Indomethacin and OKY-046 did not, however, reduce 125I-EGF binding. Thus, in GEC, the basal synthesis of eicosanoids enhanced EGF-induced proliferation. This effect appears to be due to enhancement of EGF receptor activation.

Spontaneous activity level and life span in rotifers: lack of support for the rate of living theory.

Spontaneous activity level was measured in rotifers of the species Asplanchna brightwelli on each day of their 5-day life span. No correlation was seen in group data between activity level and life span. However, data from individuals showed a positive correlation between life span and activity level in the latter half of the life span on 3 days out of 5. This tendency toward greater activity in the longer-lived rotifers is not consistent with the rate of living theory of aging.

Transvaginal ultrasonic assessment of endometrial growth in spontaneous and hyperstimulated menstrual cycles.

Endometrial thickness and reflectivity were assessed by transvaginal ultrasound in both spontaneous and hyperstimulated menstrual cycles. Two groups of women with ovulatory cycles were examined; women in group 1 had unexplained infertility and women in group 2 were having artificial insemination by donor because of reduced spermatogenesis; a third group (group 3) comprised women with tubal infertility undergoing hyperstimulation for in-vitro fertilization. There was no difference in endometrial thickness or reflectivity between the three groups. A basic pattern of endometrial appearance common to all cycles was found, consisting of hypoechoic, isoechoic and hyperechoic images, occurring in the early follicular, late follicular and luteal phases, respectively. In all three groups a positive correlation was found between proliferative phase plasma oestradiol concentration and endometrial thickness. Group 1 r = 0.403, P less than 0.01; group 2 r = 0.439, P less than 0.01; and group 3 r = 0.617, P less than 0.01. There was a progressive increase in endometrial growth throughout the normal cycle until a plateau was reached 5 days after the LH surge. This pattern was also seen without acceleration of the process in hyperstimulated cycles, despite supranormal levels of oestrogen. Assessment of endometrial thickness is not a useful variable in monitoring hyperstimulated cycles. No aberrations of endometrial growth or pattern were observed in the women with unexplained infertility.