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Biomedical subjects

M L Polan

Publications and source records attributed to M L Polan.

At least 55 records · Page 3Linked to original sources
PubMed1994

The effect of interleukin-1 beta (IL-1 beta) on the regulation of IL-1 receptor type I messenger ribonucleic acid and protein levels in cultured human endometrial stromal and glandular cells.

Because we hypothesize that the interleukin-1 (IL-1) system may be important in the dialogue between mother and embryo during the implantation process, we have analyzed the effect of IL-1 beta, a secretory product of the human embryo and human endometrium, on the mRNA and protein levels of IL-1 receptor type I (IL-1R tI) in the human endometrium. For this purpose, endometrial epithelial cells (EEC) and stromal cells (ESC) were isolated and cultured with progesterone (3.18 micrograms/mL) and epidermal growth factor (20 ng/mL) for 8 days in the presence or absence of hrIL-1 beta (20 pg/mL). EEC from proliferative and secretory endometrium expressed high levels of IL-1R tI mRNA compared to ESC, and these levels were not modulated by IL-1 beta. However, prostaglandin E2 levels peaked on day 4 in EEC treated with progesterone, epidermal growth factor, and IL-1 beta (208.7 +/- 92 ng/10(7) cells), whereas no prostaglandin E2 was detectable in cells not treated with IL-1 beta, indicating that these cells responded to IL-1 beta. With regard to ESC from secretory endometrium, IL-1 beta increased its own receptor mRNA levels (4 +/- 0.5-fold increase) after 8 days in culture. However, when ESC were isolated from proliferative endometrium, an up-regulation of IL-1R tI (3.5 +/- 0.5-fold increase) was observed on days 6 and 8 of culture regardless of the presence or absence of IL-1 beta. Immunoreactive IL-1R tI was identified in cultured EEC and ESC, and patterns similar to those of mRNA were observed. The constitutive presence of IL-1R tI in EEC, which was not affected by IL-1 beta, and the up-regulation of IL-1R tI mRNA by its ligand IL-1 beta in ESC isolated during the luteal phase suggest a role for the IL-1 system in human implantation.

Endometrium↗
PubMed1994

Interleukin-1 system in the materno-trophoblast unit in human implantation: immunohistochemical evidence for autocrine/paracrine function.

Interleukin-1 receptor type I, IL-1 beta, IL-1 receptor antagonist, and human macrophages were immunohistochemically localized in the villous trophoblast, maternal-trophoblast interphase, and maternal decidua during early human implantation. Immunostaining for IL-1 receptor type I was present in the syncytiotrophoblast and hyperplastic endometrial glands in the maternal decidua. Immunoreactive IL-1 beta was present in the villous cytotrophoblast, syncytiotrophoblast, intermediate trophoblast, and maternal stromal decidual cells. IL-1 receptor antagonist staining was observed in the glandular endometrium of the maternal decidua and in isolated cells located inside the chorionic villi, intervillous space, and maternal decidua. Mature human macrophages, as defined by both CD/68+ and HAM56+, were present in the chorionic villi, maternal blood of intervillous space, and maternal decidua. Co-localization studies demonstrated that macrophages in all of the reported locations also stained for immunoreactive IL-1 beta. Our results show the shared presence in maternal and embryonic tissues of this receptor-agonist-antagonist system during early human implantation. This finding supports an autocrine/paracrine role for the IL-1 system in human implantation.

Chorionic Villi↗
PubMed1994

Cytokines and reproduction.

Cytokines are important in reproduction. Interleukin-1, an established immune mediator, is one of the best-characterized members of the cytokine family. We describe what is known about the interactions between the interleukin-1 system and the hypothalamic-pituitary-gonadal, the hypothalamic pituitary-adrenal, and the hypothalamic-pituitary-thyroid axes. We also review the ovarian role of the interleukin-1 system. This cytokine has an immense and, as yet, imperfectly understood effect on the human reproductive tract. Clearly the immune system has a potential autocrine, paracrine, and endocrine role in regulating human reproductive events such as ovulation, luteinization, and implantation.

Animals↗
PubMed1993

Interleukin-1 type I receptor messenger ribonucleic acid expression in human endometrium throughout the menstrual cycle.

OBJECTIVE: To investigate the messenger ribonucleic acid (mRNA) expression of interleukin-1 (IL-1) type I receptor in the endometrial tissue of normal patients during the menstrual cycle. DESIGN: Prospective longitudinal study. SETTING: Department of Obstetrics and Gynecology, Stanford University Medical Center, Stanford, California. PATIENTS: Twenty fertile women between 19 and 41 years of age underwent hysterectomy for benign reasons (n = 9) and laparoscopy for tubal ligation (n = 11). In all cases, endometriosis was not visualized. INTERVENTIONS: Endometrial biopsy using the Novak curette was obtained at the time of surgery. MAIN OUTCOME MEASURE: Total RNA extracted from unfractioned endometrial tissue was analyzed on Northern blots by using specific complementary deoxyribonucleic acid probes. RESULTS: We found IL-1 type I receptor mRNA expression in endometrial tissue throughout the entire menstrual cycle. However, IL-1 type I receptor mRNA levels were significantly higher during both early and late luteal phases than follicular and midluteal phases. CONCLUSIONS: Our results demonstrate the presence of the IL-1 system in the human endometrium and that the receptor is regulated throughout the menstrual cycle with a 4.1-fold increased expression of the IL-1 receptor gene in the early luteal phase compared with preovulatory endometrium.

Adult↗
PubMed1993

Regulation of plasminogen activator inhibitor-1 and -2 messenger ribonucleic acid levels in human cumulus and granulosa-luteal cells.

Plasminogen activators and their inhibitors have been implicated in the process of fibrinolysis, tissue remodeling, and ovulation. Epidermal growth factor (EGF), a paracrine hormone found in the human ovary, increases plasminogen activator (PA) activity and the gene expression of PA and plasminogen activator inhibitor (PAI) in human endothelial cells and human cell lines. Gonadotropins also increase PA activity and gene expression in rat preovulatory granulosa cells. We have now analyzed the gene expression of PAI-1 and PAI-2 in uncultured human cumulus cells (CC), uncultured granulosa-luteal cells (GLC), and cultured GLC obtained from preovulatory follicles of patients undergoing assisted reproductive technologies. We also studied the effects of hCG and EGF on PAI-1 and PAI-2 mRNA levels in cultured GLC; GLC were cultured in serum-free medium for various times within 24 h with or without hCG and for 6 h with or without hCG, EGF, or EGF plus hCG. Total RNAs from CC and GLC were extracted, and blot hybridizations with 32P-labeled PAI-1, PAI-2, or 28S ribosomal RNA cDNA probes were performed. Both CC and GLC expressed PAI-1 and PAI-2 genes. In GLC, steady state levels of PAI-1 mRNA levels steadily increased within 24 h of culture, whereas PAI-2 levels peaked at 6 h of culture. PAI-1 mRNA levels were not affected by hCG or EGF at 6 h of culture, but PAI-2 mRNA levels were significantly increased by EGF at 6 h of culture. These studies demonstrate that human GLC PAI-1 and PAI-2 mRNA levels are differentially regulated and suggest that EGF may be involved in modulation of the human ovarian PA system during the periovulatory period.

Adult↗
PubMed1993

Human granulosa-luteal cells secrete parathyroid hormone-related protein in vivo and in vitro.

The presence of mRNA transcripts and/or immunoreactivity for PTH-related protein (PTHrP) in several normal mammalian tissues suggests a possible paracrine or autocrine role for this hormone. Since immunohistochemical studies of human ovary demonstrate the presence PTHrP immunoreactivity in this tissue, we wondered if ovarian follicular fluid (OVFF) might contain PTHrP. We retrospectively analyzed 28 OVFF samples obtained at ova harvest in 21 women undergoing in vitro fertilization. Fourteen samples contained significant adenylate cyclase-stimulating activity in a PTHrP-sensitive bioassay. In a subsequent prospective analysis, 41 of 45 freshly obtained OVFF samples demonstrated significant activity. This bioactivity was completely neutralized by antisera to PTHrP, but was unaffected by antisera to PTH. Fifteen OVFF samples were also analyzed in a sensitive 2-site immunoradiometric assay for PTHrP, and all 15 demonstrated significant levels of the hormone. The PTHrP levels did not correlate with the presence of an ovum in the follicle or with follicular fluid calcium. Short term (24- to 48-h) cultures of granulosa-luteal cells established from 5 OVFF samples demonstrated constitutive secretion of PTHrP using the immunoradiometric assay. Neither progesterone nor estrogen affected basal secretion. RNase protection analysis of cellular RNA prepared from cultured granulosa-luteal cells demonstrated the presence of mRNA for PTHrP in these cells. We conclude that 1) human OVFF obtained after stimulation with FSH and LH contain high concentrations of PTHrP; and 2) the granulosa-luteal cell is capable of secreting PTHrP both in vivo and in vitro.

Biological Assay↗
PubMed1993

Localization of interleukin-1 type I receptor and interleukin-1 beta in human endometrium throughout the menstrual cycle.

Previous studies in the human suggest that the interleukin-1 (IL-1) system, may be an important paracrine/autocrine mediator in local intercellular interaction in endometrial tissue. In this study we have determined that IL-1 receptor type I (IL-1R tI) is expressed at the messenger RNA (mRNA) and protein levels in glandular cells and its ligand, IL-1 beta has been localized by immunohistochemical methods in endothelial cells and isolated stromal cells in the human endometrium throughout the menstrual cycle. IL-1R tI mRNA was detected in glandular epithelium using both specific complementary DNA and complementary RNA 32P-labeled probes. Human glandular epithelium contains a 5.1-kilobase mRNA transcript throughout the complete menstrual cycle. Quantitative densitometric analysis of slot blot hybridization signals shows an increase of IL-1R tI mRNA in both early and mid-late secretory phases in comparison with the proliferative phase (P < 0.05). IL-1R tI protein was localized in endometrial glandular epithelial cells using both indirect immunofluorescence and avidin-biotin-peroxidase methods. However, more intense staining for IL-1R tI was observed in lumenal epithelial cells compared with the staining present deep in the endometrial glands. Using the same methods, IL-1 beta was detected in endothelial cells of spiral vessels and isolated stromal cells throughout the menstrual cycle, and an increased staining from proliferative to secretory phase was observed. The detection of IL-1R tI in the human endometrial epithelium and its ligand, IL-1 beta, in isolated stromal cells and endothelial cells, is another example of possible communication between the immune and reproductive systems with special relevance to human implantation.

Adult↗
PubMed1992

Ovarian steroids modulate human monocyte tumor necrosis factor alpha messenger ribonucleic acid levels in cultured human peripheral monocytes.

OBJECTIVE: To determine whether tumor necrosis factor alpha (TNF-alpha) messenger ribonucleic acid (mRNA) levels in human peripheral monocytes are regulated by ovarian steroids. DESIGN: Human granulosa-luteal cells and cultured, activated human peripheral monocytes were subjected to Northern blot analysis for TNF-alpha mRNA. SETTING: Academic research laboratory. PATIENTS: Two human female volunteers of reproductive age and in the luteal phase of the menstrual cycle. RESULTS: Human granulosa-luteal cells produce TNF-alpha mRNA. Physiological levels of progesterone (P) and estradiol (E2) modulate TNF-alpha mRNA from peripheral blood monocytes with an apparent inverse relationship between steroid concentration and TNF-alpha message. CONCLUSIONS: Progesterone and E2 at physiological concentrations regulate TNF-alpha mRNA production. The P antagonist mifepristone (RU486) and the E2 agonist/antagonist tamoxifen modulate total TNF-alpha mRNA levels, suggesting involvement of specific receptors.

Cells, Cultured↗
PubMed1992

Comparison of ovarian response in the same women with the same or different lots of human menopausal gonadotropin.

A variation in the bioactivity of different production lots of human menopausal gonadotropin (hMG) has been suggested. Therefore, we evaluated ovarian response to hMG in 14 women during three separate IVF cycles. The first two cycles were performed with the same lot (#03310027; Cycles A1 and A2); the third cycle utilized different lots of hMG (Cycle B). In all cycles, hMG was administered 3 ampules/day beginning cycle Day 3 and continued for at least 6 days. Estradiol and ultrasound evaluations were performed on Day 3, and then daily, beginning on cycle Day 8. Fourteen women completed all three cycles. There were no significant differences in baseline estradiol and ultrasound. Estradiol levels on Day 8 (A1, 754 +/- 130; A2, 700 +/- 107; B, 520 +/- 80 pg/ml, analysis of variance p greater than 0.5) and on Day 9 (A1, 1051 +/- 144; A2, 1140 +/- 155; B, 840 +/- 124 pg/ml, p greater than 0.05) were similar as well. The number of small (1.0-1.4 cm) follicles, large (greater than or equal to 1.5 cm) follicles, and total follicles (Day 8: total A1, 5.5 +/- 0.9; A2, 4.0 +/- 0.7; B, 4.5 +/- 0.9, p greater than 0.05; Day 9: total A1, 6.7 +/- 0.9; A2, 6.9 +/- 0.8; B, 6.9 +/- 0.9, p greater than 0.05) in all three cycles were also similar.(ABSTRACT TRUNCATED AT 250 WORDS)

Estradiol↗
PubMed1991

The effect of interleukin-1 on adhesion formation in the rat.

The potential role of interleukin-1 in postoperative adhesion formation was examined. Cecal abrasion gave a consistently higher adhesion score when compared with sham laparotomy, on the basis of adhesion number, density, and vascularity, and so was chosen for use in further studies. The extent of serosal bleeding during cecal abrasion did not affect adhesion scores. Intraperitoneal injection of 10 micrograms murine recombinant interleukin-1 alpha in cecally abraded animals on the day of surgery and on the following 4 days resulted in a significant increase in adhesion scores when compared with those of cecally abraded animals injected with vehicle alone. Adhesions enhanced with murine recombinant interleukin-1 alpha, which were thicker and more vascular, were equivalently enhanced at doses from 10 to 10,000 ng, implying maximal response over that range. Rats not operated on and receiving recombinant interleukin-1 alpha 2 weeks after injury had increased adhesion formation. These results demonstrate that interleukin-1 alpha may be an important short-term mediator of postsurgical adhesion formation.

Animals↗
PubMed1990

Purine levels and metabolism in human follicular fluid.

Adenosine (ADO) in low micromolar levels and hypoxanthine (HYP) in millimolar levels have been shown to inhibit maturation of cumulus-enclosed oocytes. To determine the effect of ovarian stimulation with gonadotrophins on follicular purine metabolism, we measured ADO, HYP, inosine (INO), adenine (ADE) and cyclic AMP (cAMP) levels in follicular fluid (FF) from natural (n = 7) or human menopausal gonadotrophin/human chorionic gonadotrophin (HMG/HCG)-stimulated (n = 35) cycles. Purines were extracted immediately (natural cycles) or within 30 min of recovery (HMG/HCG cycles) and analysed by high pressure liquid chromatography (HPLC). The concentration of all ADE purines in FF was in the low micromolar range (1-20 microM); cAMP levels were markedly increased (greater than 100 microM) in FF of HMG/HCG-treated patients. While ADO levels were within the range effective for inhibition of oocyte maturation, those of HYP were not. No correlation was found between purine levels in FF and ovum maturation. Purine levels in FF of natural cycles were uniformly lower than those of stimulated cycles. Significant conversion of 5'-AMP into ADO, ADO into INO and INO into HYP occurred within 1 h when FF was incubated at 25 but not at 4 degrees C. These purine levels in human FF confirm our previous findings with bovine FF and suggest a possible role of ADO, but not of HYP, in the inhibition of oocyte maturation in the human.

Adenine↗
PubMed1990

Cultured human luteal peripheral monocytes secrete increased levels of interleukin-1.

Interleukin-1 (IL-1), an immune monokine secreted by activated macrophages and monocytes, appears to be intimately involved in endocrine phenomena. It is well established that IL-1 mediates a number of immune and inflammatory responses. In addition, the production of IL-1 appears to be modulated by the gonadal estradiol and progesterone, leading to this study of IL-1 secretion by cultured peripheral monocytes isolated at varying times during the menstrual cycle and pregnancy. Circulating peripheral monocytes were isolated from eight women undergoing human menopausal gonadotropin-stimulated ovulation induction during the late follicular phase just before hCG administration and from eight similarly stimulated women on the 12th day after hCG administration. Peripheral monocytes were also isolated from six women during both second and third trimester pregnancy. After 48 h in culture, conditioned media were assayed for IL-1 bioactivity using the D1O lymphocyte proliferation assay. Significantly more IL-1 bioactivity was secreted by monocytes isolated in the luteal phase of the cycle (52.4 +/- 17.5 IU/mL) compared to that in late follicular phase (5.2 +/- 0.9 IU/mL; P = 0.001) or third trimester of pregnancy (7.0 +/- 1.5 IU/mL; P = 0.006). Thus, peripheral monocyte IL-1 secretion appears to be increased by luteal levels of progesterone, although further elevation of progesterone during pregnancy returns IL-1 levels to the preovulatory baseline. In addition, basal body temperatures obtained in four women during third trimester pregnancy were 98 F or less. Thus, IL-1 secretion from cultured monocytes appears to increase with luteal concentrations of progesterone and decrease to preovulatory levels at higher concentrations of the steroid during pregnancy, which may account for the dissociation of high progesterone levels and elevated basal body temperature during late pregnancy.

Adult↗
PubMed1990

Human follicular fluids contain tissue macrophages.

Tissue cells obtained from the follicular fluids of women undergoing in vitro fertilization and embryo transfer were found to contain interleukin-1 beta mRNA by Northern analysis. Since macrophages are known to produce interleukin-1 beta, we examined the follicular fluids of 20 women undergoing in vitro fertilization as well as tissue sections of normal human ovary for the presence of macrophages and monocytes. Although granulosa-luteal cells predominate in follicular fluid, we found that resident macrophages and monocytes comprise 5-15% of human follicular tissue cells. In addition, we observed that macrophages are present in the human ovarian follicle as well as in the corpus luteum.

Blotting, Northern↗
PubMed1989

Absence of a direct effect of recombinant interleukins and cultured peritoneal macrophages on early embryonic development in the mouse.

On the basis of their recent studies, several researchers have suggested that the infertility associated with mild endometriosis is due to the alteration of peritoneal fluid, resulting in impairment of the viability of gametes or embryos. Elevated numbers of macrophages and lymphocytes have been reported in the peritoneal fluid of patients with endometriosis. Interleukin-1 (IL-1) a major product of activated macrophages and Interleukin-2 (IL-2), a product of most activated T-cells, have been postulated to play a role in the infertility associated with this disease, possibly by acting as direct embryotoxic agents. We have examined the effect of purified recombinant IL-1 and IL-2, which are not species-specific, on in vitro development of mouse embryos. Both interleukins had no effect on development to the blastocyst stage or on early stages of implantation, as measured in vitro by attachment and outgrowth of blastocysts to fibronectin-coated dishes. Moreover, co-culture of mouse embryos with activated human peritoneal macrophages had no effect on embryogenesis. We conclude that neither IL-1, nor other products of human macrophages activated by lipopolysaccharide, nor IL-2 are directly toxic to early mouse embryonic development.

Animals↗
PubMed1989

Progesterone and estradiol modulate interleukin-1 beta messenger ribonucleic acid levels in cultured human peripheral monocytes.

The relationship between the endocrine system and immune monokines, such as interleukin-1 (IL-1), is of increasing interest. IL-1, a protein secreted by peripheral monocytes and tissue macrophages, mediates a wide variety of immune responses, and its production appears to be inversely related to the level of gonadal steroids. In this report, we have investigated the relationship between estradiol and progesterone concentrations and the level of IL-1 beta mRNA in cultured human peripheral monocytes and pelvic macrophages. Human peripheral monocytes, isolated during the luteal phase of the menstrual cycle, were activated with lipopolysaccharide (10 micrograms/mL). Cellular RNA was isolated and analyzed by Northern analysis using an 800-basepair IL-1 beta cDNA probe. Hybridization with 32P-labeled probe showed maximal levels of IL-1 beta mRNA occurring between 3 and 7 h of culture. Cultures of lipopolysaccharide-activated human peripheral monocytes incubated for 3-6 h with increasing amounts of progesterone or estradiol (0-10(-5) M) in the presence of either 5% fetal calf serum or 0.1% BSA demonstrated an inverse relationship between IL-1 beta mRNA levels and steroid concentration. In both cases, IL-1 beta mRNA levels decreased by 80-90% as the progesterone concentration increased to 10(-5) M and by 70-90% as the estradiol concentration increased similarly. A similar 80% decrease in IL-1 beta mRNA was observed with peritoneal macrophages incubated with increasing amounts of progesterone. This reciprocal relationship between IL-1 beta mRNA and gonadal steroids may have important ramifications in reproductive biology for both embryonic implantation and fetal survival as well as for clinically relevant changes in bone mass.

Adult↗
PubMed1988

Human menopausal gonadotropins: a justifiable therapy in ovulatory women with long-standing idiopathic infertility.

Long-standing infertility of unknown etiology is currently an indication for in vitro fertilization. Because of the prolonged waiting time, expense, and operative risk, we have treated a group of women with empiric human menopausal gonadotropins/human chorionic gonadotropin before in vitro fertilization. A study group of 97 couples with well-documented, long-standing idiopathic infertility received up to four cycles of empiric human menopausal gonadotropin therapy before in vitro fertilization. This group was compared with a control group of 48 couples who did not receive human menopausal gonadotropin before in vitro fertilization. The 12 (12.4%) conceptions and eight (8.2%) term births resulting from empiric human menopausal gonadotropin therapy in the study group was significantly higher than the number of spontaneous conceptions and births (1%) in the study group (p less than 0.003) and was also greater than the spontaneous conception and birth rate (4%) in the control group (p less than 0.07). The conception rate with empiric human menopausal gonadotropin (12.4%) compared favorably with that after in vitro fertilization (7.9%) and 8 of 12 (66.7%) of empiric human menopausal gonadotropin conceptions resulted in term births, compared with two of seven (28.6%) of the in vitro fertilization conceptions. Preovulatory serum estradiol levels were similar for all study patients undergoing empiric human menopausal gonadotropin therapy, control patients undergoing in vitro fertilization, and patients receiving empiric human menopausal gonadotropin during conception cycles. It therefore appears appropriate to offer a 4-month trial of empiric human menopausal gonadotropin to that specific group of couples with long-standing idiopathic infertility.

Adult↗
PubMed1988

Spironolactone does not affect in vitro androgen secretion by human granulosa-luteal cells.

Although spironolactone has been documented to decrease peripheral serum androgen levels in women treated for androgen excess of ovarian origin, the site of action does not appear to be the granulosa cell. In vitro cell culture studies using human granulosa-luteal cells isolated from women undergoing ovarian stimulation for IVF have shown no affect on T, delta 4A, or DHT secretion in the presence of clinically therapeutic levels of spironolactone (0-10(-6) M). In addition, neither P nor E2 production was affected by increasing concentrations of spironolactone.

Androgens↗