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Biomedical subjects

L C Giudice

Publications and source records attributed to L C Giudice.

At least 109 records · Page 6Linked to original sources
PubMed1992

Local anesthesia with conscious sedation for laparoscopic intrafallopian transfer.

Local anesthesia with conscious sedation is well accepted by patients and provides scheduling flexibility, cost containment, patient safety, and ease of recovery. We believe the technique should be offered to selected patients undergoing intrafallopian transfer. By adhering to specific guidelines for surgical technique and monitoring, the procedure is a safe and acceptable alternative to general anesthesia for laparoscopic intrafallopian transfers.

Adult↗
PubMed1991

Human granulosa contain messenger ribonucleic acids encoding insulin-like growth factor-binding proteins (IGFBPs) and secrete IGFBPs in culture.

OBJECTIVE: To determine if luteinizing human granulosa cells contain messenger ribonucleic acid (mRNAs) encoding insulin-like growth factor-binding protein (IGFBPs) and if cultured granulosa secrete IGFBPs into conditioned medium (CM). DESIGN: Northern analysis, using IGFBP-specific complementary deoxyribonucleic acid probes, was used to detect granulosa-derived IGFBP mRNAs. Western ligand blot analysis of CM was used to detect IGFBPs secreted by granulosa cultures with and without human chorionic gonadotropin (hCG). SETTING: Granulosa cells were obtained from the In Vitro Fertilization (IVF) Program at Stanford University, a private teaching institution. PATIENTS, PARTICIPANTS: Patients undergoing IVF for tubal disease. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Transcripts of IGFBP mRNA and IGFBPs secreted into CM were detected by autoradiography of Northern and Western ligand blots, respectively. RESULTS: Transcripts of IGFBP-3, IGFBP-2, and IGFBP-1 mRNA were detected in human luteinizing granulosa. Cultured granulosa secreted IGFBPs with molecular weights corresponding to IGFBP-3, IGFBP-2, and IGFBP-1, and the latter two IGFBPs increased with 10 ng/mL hCG. A 24 kd IGFBP was noted, which may be newly characterized IGFBP-4. CONCLUSIONS: These data show that luteinizing human granulosa cells express mRNAs encoding three IGFBPs, secrete IGFBPs into culture medium, and that production of at least two of the IGFBPs is hCG-dependent, further supporting a role for the IGF system in human folliculogenesis.

Blotting, Northern↗
PubMed1991

Characterization of the steroid-dependence of insulin-like growth factor-binding protein-2 synthesis and mRNA expression in cultured human endometrial stromal cells.

The insulin-like growth factors (IGF-I and -II) are believed to be important in endometrial differentiation and blastocyst nidation, and proteins that regulate IGF action (IGF-binding proteins, IGFBPs) are hormonally regulated in endometrium during the menstrual cycle. To characterize further steroid-dependence of the IGFBPs, we established endometrial stromal cells in culture in the absence and presence of oestradiol (E2) and progesterone (P) and examined the conditioned medium for IGFBPs by Western ligand blot analysis. Stromal cells constitutively synthesized IGFBP-3, IGFBP-2, a 27 kd, and a 24 kd IGFBP. In the presence of E2 and P, a 10- to 15-fold increase in IGFBP-2 was detected in the conditioned medium beginning after about 7 days in culture, when cells decidualized and steroid-mediated prolactin secretion began. Withdrawal of steroids resulted in a marked decrease in IGFBP-2, comparable to control levels, and cells increased their IGFBP-2 production when rechallenged with E2 and P. Total RNA was isolated from stromal cells, and Northern blot analysis using a cDNA probe specific for IGFBP-2 revealed differential expression of a 1.4 kb mRNA transcript in steroid-treated compared to control cells. The effects of progestational agents alone on IGFBP synthesis was also examined. Progesterone, medroxyprogesterone acetate and norethindrone all stimulated IGFBP-2 synthesis 12- to 15-fold compared to controls, and a progesterone receptor antagonist, RU 486, blocked the stimulatory effect of progesterone. IGFBP-2 synthesis was increased two-fold above controls by 17-alpha-hydroxyprogesterone, and RU 486 alone and hydrocortisone were without effect. Identification of IGFBP-2 in conditioned medium was made using IGFBP-specific antiserum. These data show that (a) endometrial stromal cells synthesize and secrete IGFBP-2, (b) IGFBP-2 protein synthesis is controlled by steroid hormones, (c) P, by interacting with its receptor, modulates IGFBP-2 synthesis and (d) expression of IGFBP-2 mRNA is controlled by sex steroids.

Autoradiography↗
PubMed1991

Insulin-like growth factor-II (IGF-II) and IGF binding proteins in human endometrium.

The insulin-like growth factor autocrine/paracrine system is believed to play a role in steroid-mediated endometrial differentiation. It is constituted of the mitogenic peptides (IGF-I and IGF-II), membrane receptors, and a family of high-affinity binding proteins (IGFBPs) that regulate the actions of the IGFs at their target cells. We have investigated expression of the mRNAs encoding the three major IGFBPs (IGFBP-1, IGFBP-2, and IGFBP-3) in human endometrium and have found, by Northern analysis, differential expression of all three mRNAs in secretory compared to proliferative endometrium, different steroidal milieux. IGF-II mRNAs were also detected in secretory endometrium. Finally, we found that human endometrial stromal cells in culture synthesize and secrete IGFBP-2 and IGFBP-3, and that the synthesis of IGFBP-2 is regulated by steroid hormones.

Autoradiography↗
PubMed1991

Maternal insulin-like growth factor-binding protein messenger ribonucleic acid during rat pregnancy.

Pregnancy is associated with rapid growth of maternal and fetal tissues. Insulin-like growth factors (IGF-I and -II) have roles in mediating both fetal and placental growth. In this study serum IGFs and IGF-binding proteins (IGFBPs) were characterized, IGFBP protease activity was quantified, and hepatic IGFBP-1, -2, -3, and -4 mRNA were investigated throughout rat pregnancy. IGF-I in maternal serum was elevated (P less than or equal to 0.001) on days 5 and 10 (d5 and d10) of gestation compared to levels in nonpregnant controls (NP), but was significantly decreased below NP levels (P less than or equal to 0.001) after d10 of pregnancy. Serum IGF-II levels were unaffected by pregnancy. Using Western ligand blotting (WLB), six IGFBP bands were visualized in NP, d5, and d10 pregnancy rat sera. At 15 and 20 days gestation, the IGFBP-3 bands were no longer detectable by WLB. Using an in vitro IGFBP protease assay, sera from rats at 15 and 20 days gestation proteolyzed 63 +/- 4% and 81 +/- 5% of recombinant human IGFBP-3, respectively. Regression analyses demonstrated that serum IGF-I was positively correlated with serum IGFBP-3 (r2 = 0.73; P = 0.001), whereas serum IGFBP-3 (r2 = -0.85; P = 0.001) and serum IGF-I (r2 = -0.78; P = 0.001) were negatively correlated with serum protease activity. In addition, no change was observed in liver IGFBP-3 mRNA during pregnancy, further suggesting that protease activity is primarily responsible for the decrease in serum IGFBP-3. However, IGFBP-1 and -4 mRNA levels were increased 3- to 11-fold after d5 of gestation. The hormonal and/or metabolic regulators of hepatic IGFBP-1 and -4 expression during rat pregnancy remain to be determined.

Animals↗
PubMed1991

Insulin-like growth factor binding proteins in human endometrium: steroid-dependent messenger ribonucleic acid expression and protein synthesis.

The insulin-like growth factor (IGF) autocrine/paracrine system is believed to play a role in endometrial differentiation and trophoblast growth. The IGFs bind with high affinity to a family of binding proteins (IGFBPs) that regulate the actions of the IGFs at their target cells. IGFBP-1, one of three well-characterized IGFBPs in humans, has been shown by numerous investigators to be present in secretory but not proliferative endometrium; however, no information is available with regard to two other human IGFBPs, IGFBP-2 and IGFBP-3, in normal human endometrium. We have examined expression of the messenger RNAs (mRNAs) encoding IGFBP-2 and IGFBP-3 in human proliferative endometrium [under the influence of estradiol (E2)] compared to secretory endometrium (under the influence of E2 and progesterone). Using a complementary DNA probe specific for IGFBP-2, by Northern analysis, expression of a 1.4 kilobase mRNA was found to be differentially expressed in secretory compared to proliferative phase endometrium. Using a complementary DNA probe encoding IGFBP-3, a 2.4 kilobase mRNA was found in endometrium and was also found to be differentially expressed in secretory compared to proliferative phase endometrium. Endometrial stromal cells were established in culture and revealed constitutive synthesis and secretion of IGFBP-2 and IGFBP-3 into the conditioned medium (CM), as detected by Western ligand blot analysis of the CM. Identification of the IGFBPs in the CM was made using IGFBP-2 and IGFBP-3 specific antiserum. In the presence of E2 and progesterone, there was an enhancement in the amount of IGFBP-3 and a marked enhancement of IGFBP-2 in the conditioned media, suggesting sex steroid-dependence of IGFBP-2 and, to a lesser extent, IGFBP-3 protein synthesis. Western ligand blot analysis of IGFBPs in serum throughout the menstrual cycle revealed no hormonal dependence in the serum IGFBPs, suggesting local action of the IGFBPs in endometrium. These data thus show that mRNAs encoding IGFBP-2 and IGFBP-3 are expressed in human endometrium, that their expression is differentially regulated in secretory compared to proliferative phase endometrium (different steroidogenic environments), and that the synthesis of both IGFBP-2 and IGFBP-3 proteins is regulated by steroid hormones.

Autoradiography↗
PubMed1991

A simple assay for proteolysis of IGFBP-3.

An assay for assessing the proteolysis of IGFBP-3 by a biological fluid or protein has been developed using 125IhIGFBP-3 recombinantly derived from Escherichia coli as a substrate indicator. The labelled substrate is incubated at 37 degrees in the presence of the agent being tested and proteolysis is visualized by SDS-PAGE and autoradiography. The assay showed that the pregnancy associated protease is Ca++ dependent. The substrate is also proteolyzed by rat and mouse pregnancy sera, making the assay applicable to studies in those animal systems.

Animals↗
PubMed1990

Identification of insulin-like growth factor-binding protein-2 (IGF-BP-2) and a low molecular weight IGF-BP in human seminal plasma.

Three distinct binding proteins for the insulin-like growth factors (IGF-BPs) have recently had their cDNAs cloned and sequenced. We have evaluated IGF-BP expression in normal, oligospermic and azospermic human seminal plasma (SP), and have identified IGF-BP-2 in all samples. Additionally, SP contains a non-glycosylated low molecular weight IGF-BP (24K), which does not appear to be structurally or immunologically related to IGF-BP-1, -2 or -3.

Base Sequence↗
PubMed1990

Insulin-like growth factor binding proteins in maternal serum throughout gestation and in the puerperium: effects of a pregnancy-associated serum protease activity.

The cDNAs encoding three major insulin-like growth factor-binding proteins (IGFBPs) have been cloned and sequenced. We have examined, by Western ligand blotting, the profiles of these binding proteins in human female serum in the normal menstrual cycle, throughout pregnancy, and during the postpartum period. There was no change in the serum profile of any of the binding proteins in early pregnancy compared to that in the secretory phase of the menstrual cycle. However, there was a marked decrease in circulating levels of the main serum IGFBP, IGFBP-3, after 6 weeks of gestation, continuing progressively to term and returning to nonpregnant levels by 5 days postpartum. IGFBP-2 decreased steadily throughout gestation. In contrast, IGFBP-1 levels were found to rise by the second trimester. Endoglycosidase-F digestion did not enhance detection of IGFBP-3 by ligand blotting. Immunoprecipitations with two separate antibodies against IGFBP-3 and IGFBP-2, followed by Western ligand blotting, confirmed the marked decrease in IGFBP-3 levels after 6 weeks of gestation and the more gradual decrease in IGFBP-2. In contrast, immunoprecipitations with IGFBP-1 monoclonal antibodies confirmed the increase in IGFBP-1 during gestation. Endogenous serum IGFs were separated from serum IGFBPs by acid chromatography, and an 80% decrease in total IGF-binding activity in the IGFBP fraction of chromatographed pregnancy vs. nonpregnancy serum was detected by charcoal absorption assay. Furthermore, immunoprecipitations of IGF affinity cross-linked IGFBP fractions with IGFBP-3-specific antiserum confirmed a marked diminution of IGFBP-3 in pregnancy compared to nonpregnancy serum, and revealed, only in pregnancy serum, the concomitant appearance of a band with a mol wt of 34K and three less intense bands with mol wt between 20-26K on sodium dodecyl sulfate gels. Incubation of nonpregnancy serum with 6-week pregnancy serum at 37 C for 5 h, followed by Western ligand blotting, showed only a slight reduction in the amount of IGFBP-3 in the mixture compared to that in controls. However, incubation of term pregnancy with nonpregnancy serum at 37 C for 5 h revealed a marked reduction of IGFBP-3 in the mixture. When iodinated recombinant IGFBP-3 was incubated with term pregnancy serum under the same conditions, the appearance of a 29K protein was identified by gel electrophoresis and autoradiography, along with three less intense bands with mol wt between 17-22K.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗
PubMed1990

Identification of insulin-like growth factor-binding protein-3 (IGFBP-3) and IGFBP-2 in human follicular fluid.

Two major insulin-like growth factor-binding proteins, IGFBP-2 and IGFBP-3, were identified by Western ligand blotting of human follicular fluid (FF) aspirated from luteinizing follicles during ovarian stimulation with human menopausal gonadotropin (LH and FSH). IGFBP-1 was also present in FF, and an IGFBP with a mol wt of 24000 daltons, present in serum, appeared to be absent in FF. Identification of IGFBP-1, IGFBP-2, and IGFBP-3 was confirmed using IGFBP-specific antisera. IGF-I and IGF-II levels were measured in patient serum and FF at the time of oocyte harvest, and serum levels of both peptides were consistently higher than FF levels. The finding of three major serum IGFBPs in human FF suggests that IGF regulation in the ovary is probably under complex control by this set of regulatory BPs.

Antibodies, Monoclonal↗
PubMed1989

Expression of two insulin-like growth factor-binding proteins in a human endometrial cancer cell line: structural, immunological, and genetic characterization.

Insulin-like growth factors (IGFs) bind with high affinity to specific proteins in human serum, cerebrospinal fluid (CSF), and amniotic fluid. In serum, IGFs are bound to a complex with an apparent mol wt of 150K in which an acidstable binding protein of 53K, termed BP-53, is found. In amniotic fluid, a different binding protein with an apparent mol wt of 25K, termed hBP-25, has been identified. This binding protein is secreted by endometrial cells and has been shown to block the binding of IGFs to their membrane receptors and inhibit the mitogenic action of the IGFs on human choriocarcinoma cells. It has been proposed that hBP-25 modulates the action of IGFs on endometrial tissue, especially during pregnancy. We have identified two binding proteins produced by an endometrial adenocarcinoma, HEC 1A, neither of which is related structurally or genetically to hBP-25. One is present in two glycosylated forms with apparent mol wt of 37K and 40K, both of which are immunoprecipitated with an antiserum made to BP-53. Both forms are reduced to a core protein of 30K upon digestion with endoglycosidase-F. Furthermore, BP-53-specific RNA was detected in HEC 1A cells. A second binding protein with an apparent mol wt of 32K was also detected and did not increase in size upon treatment with endo glycosidase F. These two binding proteins were partially purified by a combination of wheat germ agglutinin and IGF affinity chromatography, and polyclonal antibodies were generated. The polyclonal antiserum specifically recognizes a 32K protein in human CSF, suggesting that the HEC 1A cells produce the same binding protein that is predominant in CSF. The findings suggest that IGF action in the endometrium may be modulated by more than a single binding protein, and that at least three structurally distinct human IGF binding proteins exist.

Blotting, Northern↗
PubMed1988

Cryopreservation of embryos and ova.

Development of techniques for cryopreservation of embryos of several species, principally the mouse, laid the foundation for cryopreservation of human embryos. As IVF has become more widely available and the need for the cryopreservation of human embryos has become apparent, pressure for technical development has increased. The ideal method would be simple, inexpensive, and effective. The most effective method for cryopreservation of early human embryos, such as those at the 1-cell pronuclear stage and up to the 4-cell stage, now appears to be stepwise cooling in 1,2-propanediol with sucrose in plastic ministraws. The preferred method for intermediate stage embryos uses DMSO with cooling and thawing at slow rates in a programmed biologic freezer. For the human blastocyst, slow cooling in glycerol and rapid thawing is the only method reported with survival rates comparable to those achieved for intermediate stage embryos using DMSO. The rates of survival from freezing and thawing blastocysts are not sufficiently high, however, to justify the losses associated with prolonged in vitro incubation. Even at the current level of technical achievement, cryopreservation of human embryos provides the clearest opportunity to improve the clinical results obtained with IVF. Research now underway in the modification of methods for vitrification and ultrarapid freezing holds promise for both simplification of technology and improvement of outcome. In view of legal and ethical considerations involved in embryo preservation, the desirability of ova preservation is widely accepted. Although a small number of human unfertilized mature ova have been cryopreserved using various methods, success rates are still low. Methods for the cryopreservation of eggs should be developed, but these methods probably should be proved by animal experiments to be safe, especially with regard to genetic damage, before a policy of transfer of embryos derived from frozen-thawed human ova is applied on a large scale.

Algorithms↗
PubMed1986

Serum levels of CA-125 in patients with endometriosis: a preliminary report.

Seven out of 8 patients with endometriosis demonstrated levels of CA-125 antigen above 35 U/ml. None of 15 patients with other benign gynecologic diagnoses demonstrated elevated levels. This antigen has been proposed as a tumor marker for epithelial carcinoma and other gynecologic neoplasms. However, it cannot be used to differentiate clinically between cancer and endometriosis.

Adolescent↗