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Mary Lake Polan

Publications and source records attributed to Mary Lake Polan.

At least 19 recordsLinked to original sources
PubMed2007

The role of neutrophil elastase in elastin metabolism of pelvic tissues from women with stress urinary incontinence.

OBJECTIVE: Altered elastin metabolism is implicated in pelvic floor disorders. We studied neutrophil elastase (NE) and matrix metalloproteinase (MMP) activities in vaginal tissues from premenopausal women with stress urinary incontinence (SUI). METHODS: Elastase and NE activities in vaginal tissues were assessed. Protein and mRNA expressions were determined by RT-PCR and Western blot. Total elastin and collagen contents were evaluated. To compare the relative elastolytic effect of NE and MMP-2, we used their respective antibodies to immunoprecipitate these proteins from vaginal fibroblast extracts prior to assessing elastase activity. RESULTS: Elastase activity in vaginal wall tissues was significantly higher in the secretory compared to the proliferative phase. NE mRNA and protein expressions were similar between control and SUI tissues from the secretory phase. However, NE activity in the SUI tissues was higher compared to control tissues. The mRNA expression of alpha-1 antitrypsin (ATT) was higher in control tissues from the proliferative phase compared to those from the secretory phase, while no difference was observed in SUI tissues between either phase. Protein expression of the active form of ATT was decreased in SUI tissues compared to controls during the secretory phase. Anti-NE antibody reduced total elastase activity by 60-70%, compared to less than 20% reduction with anti-MMP-2 antibody. CONCLUSION: During the secretory phase, elastolytic activity is increased in pelvic tissues from women with SUI, through an increase in NE activity and a concurrent decrease in ATT expression. The serine protease, NE, appears to be a more significant modulator of elastase activity compared to MMP-2.

Adult↗
PubMed2006

Do extracellular matrix protein expressions change with cyclic reproductive hormones in pelvic connective tissue from women with stress urinary incontinence?

BACKGROUND: To evaluate differential expression of transforming growth factor (TGF-beta1), latent transforming factor-binding proteins (LTBP-1, LTBP-2) and elastin microfibril components (fibrillin-1 and fibrillin-2) in vaginal tissue from women with stress urinary incontinence (SUI). METHODS: In this case-control study, vaginal tissue from women in both phases of the menstrual cycle was obtained. Messenger RNA (mRNA) expressions of LTBP-1, LTBP-2, fibrillin-1, fibrillin-2 and TGF-beta1 were determined by relative real-time quantification PCR. Tissue localization was analysed by immunohistochemistry, and semiquantitative protein expression was evaluated by Western blot analysis. RESULTS: Vaginal wall fibroblasts synthesized all proteins tested. LTBP-1, LTBP-2 and TGF-beta1 co-localized with elastin microfibrils, fibrillin-1 and fibrillin-2 in the extracellular matrix. LTBP-1 mRNA and protein expressions were higher in control versus women affected with SUI in the proliferative phase (P = 0.04), while in the secretory phase, mRNA expression in cases was higher (P = 0.04). Fibrillin-1 mRNA was higher in women affected by SUI versus controls in both phases, but no statistical differences in fibrillin-1 protein expression were observed between the two groups in either phase. LTBP-2 and TGF-beta1 mRNA expressions showed the same trends as LTBP-1. CONCLUSION: LTBP-1, LTBP-2, TGF-beta1, fibrillin-1, and fibrillin-2 expressions are hormonally regulated in vaginal wall fibroblasts and differ in women affected by SUI when compared to controls. These data suggest a mechanism to regulate TGF-beta1 activity in pelvic connective tissue.

Case-Control Studies↗
PubMed2006

Regulation of cyclooxygenase activity in cultured endometrial stromal cells by granulocyte-macrophage colony-stimulating factor.

OBJECTIVE: To assess the ability of granulocyte-macrophage colony-stimulating factor (GM-CSF) to regulate cyclooxygenase (COX) enzyme activity and prostaglandins (PGs) synthesis, specifically PGE2 production in stromal cells, neither of which have been addressed in the literature. DESIGN: Prospective study. SETTING: Department of obstetrics and gynecology at a university hospital. PATIENT(S): Human luteal phase endometrium was obtained from surgical specimens (n = 6) for clinical indications. INTERVENTION(S): Confluent stromal cells were stimulated with GM-CSF. MAIN OUTCOME MEASURE(S): Expression of COX mRNA, COX enzyme activity, and PGE2 level in cultured stromal cells. RESULT(S): Confluent stromal cell cultures treated with P and E2 for 9 days were stimulated with GM-CSF. After treatment of 12 hours, low-dose GM-CSF (0.001-0.1 ng/mL) increased COX-2 mRNA levels in stromal cell, whereas high dose GM-CSF (1-100 ng/mL) decreased COX-1 and COX-2 mRNA levels. After treatment of 48 hours, low concentrations of GM-CSF (0.001-0.1 ng/mL) increased total COX and COX-2 enzyme activity, whereas high concentrations of GM-CSF (1-100 ng/mL) inhibited COX and COX-2 activity; The PGE2 levels decreased by 31% to 393.3 pg/mL (P < .05) with concentrations of GM-CSF increasing from 1 ng/mL to 100 ng/mL. CONCLUSION(S): There appeared to be a biphasic pattern of COX-2 enzyme response to GM-CSF with low concentrations increasing activity and high concentrations inhibiting activity. It is possible that GM-CSF may provide critical regulation of PG production in the preimplantation period.

Cells, Cultured↗
PubMed2005

Microarray analysis of differentially expressed genes in vaginal tissues from women with stress urinary incontinence compared with asymptomatic women.

BACKGROUND: The pathophysiology of pelvic floor dysfunction resulting in stress urinary incontinence (SUI) in women is complex. Evidence suggests that there is also a genetic predisposition towards SUI. We sought to identify differentially expressed genes involved in extracellular matrix (ECM) metabolism in vaginal tissues from women with SUI in the secretory phase of menses compared with asymptomatic women. METHODS: Tissue samples were taken from the periurethral vaginal wall of five pairs of premenopausal, age-matched SUI and continent women and subjected to microarray analysis using the GeneChip Human Genome U133 oligonucleotide chip set. RESULTS: Extensive statistical analyses generated a list of 79 differentially expressed genes. Elafin, keratin 16, collagen type XVII and plakophilin 1 were consistently identified as up-regulated ECM genes. Elafin, a serine protease inhibitor involved in the elastin degradation pathway and wound healing, was expressed in pelvic fibroblasts and confirmed by Western blot, quantitative competitive PCR and immunofluorescence cell staining. CONCLUSIONS: Genes involved in elastin metabolism were differentially expressed in vaginal tissue from women with SUI, suggesting that elastin remodelling may be important in the molecular aetiology of SUI.

Adult↗
PubMed2005

Exogenous granulocyte-macrophage colony-stimulating factor promotes follicular development in the newborn rat in vivo.

BACKGROUND: Expression and selective cellular localization of granulocyte-macrophage colony-stimulating factor (GM-CSF) and its receptor in ovarian tissue imply an autocrine/paracrine role in ovarian function. Evidence indicating a functional role for GM-CSF in ovarian follicular cell function has been provided by studies with GM-CSF knockout (GM-/-) mice, which suggest that GM-CSF influences events associated with murine follicular maturation. METHODS: Immature female rats were treated with GM-CSF, FSH or saline for 5 or 10 days. Ovaries were collected for histologic examination and immunostaining determination of CYP17, a theca cell marker. In addition, ovarian section slides were evaluated by immunofluorescence for CD45, an ovarian leukocyte marker. To investigate the possible mechanism of GM-CSF action on follicular development, theca-interstitial cells (T-I) were separated and cultured. Cells were treated with increasing concentrations of GM-CSF, then evaluated for CYP17 mRNA and protein expression assays. RESULTS: After 10 days of treatment with GM-CSF, the number of small preantral and large preantral follicles was significantly increased compared with the control group (P < 0.05). Similarly, treatment with FSH increased the number of small preantral and large preantral follicles (P < 0.05). CD45 expression measured by immunofluorescence was not different in the three groups, indicating that the distribution of leukocytes was unchanged. In addition, CYP17 was increased in the T-I cells both in vivo and in vitro after GM-CSF treatment. CONCLUSION: The present results suggest that GM-CSF may play a significant role in follicular development.

Animals↗
PubMed2005

Preliminary experience with low concentration of granulocyte-macrophage colony-stimulating factor: a potential regulator in preimplantation mouse embryo development and apoptosis.

PURPOSE: To investigate the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the development of preimplantation mouse embryos. METHODS: Mouse 2-cell embryos were collected and cultured in P-1 medium supplemented with GM-CSF at different concentrations. Using reverse transcription-polymerase chain reaction, expression Bcl-2 and Bax mRNA in blastocyst were evaluated in the GM-CSF group and control group. Apoptosis detection was performed using the in situ apoptosis detection kit in mouse blastocysts. The statistical significance of the data was analyzed using t-test and chi-square test. RESULTS: The development of blastocyst increased to 89% in the addition of GM-CSF (0.125 ng/mL), compared to controlled group (80%). The number of cells staining for apoptosis was lower in GM-CSF group than that in the control group. Bcl-2 expression was found to be upregulated in blastocysts in the GM-CSF supplemented group compared to the control group. CONCLUSION: These results suggest that GM-CSF might be an important regulator in embryo development.

Animals↗
PubMed2005

Elastin metabolism in pelvic tissues: is it modulated by reproductive hormones?

OBJECTIVE: The purpose of this study was to investigate the effect of relaxin on extracellular matrix protein expression in pelvic fibroblasts that were cultured from women with stress urinary incontinence compared with asymptomatic control subjects. STUDY DESIGN: Periurethral vaginal wall fibroblasts from premenopausal women with stress urinary incontinence and continent women (in both the proliferative and secretory phase of the menstrual cycle) were stimulated with increasing concentrations of relaxin (0-500 ng/mL). The supernatant was sampled for matrix metalloproteinase-2 and -9 by zymography. Tissue inhibitors of metalloproteinase-1 and -2 and alpha-1 antitrypsin were evaluated with Western blot. Total elastase activity was measured by generation of free amino groups from succinylated elastin. Increasing concentrations of alpha-1 antitrypsin were added to cell lysate to evaluate total elastase activity inhibition. RESULTS: Proliferative-phase stress urinary incontinence fibroblasts demonstrated an increase in matrix metalloproteinase-2 and no change in matrix metalloproteinase-9 and tissue inhibitors of metalloproteinase-1 and -2 expressions with increasing relaxin concentrations. Cells from control subjects showed increased expression of matrix metalloproteinase-2 and -9, but no change in tissue inhibitors of metalloproteinases. Secretory-phase stress urinary incontinence fibroblasts showed no response in matrix metalloproteinase or tissue inhibitors of metalloproteinase expressions with relaxin stimulation. Secretory-phase control fibroblasts reacted by increasing matrix metalloproteinase-2 and -9 and tissue inhibitors of metalloproteinase-2. With respect to total elastase activity and alpha-1 antitrypsin expression, increasing doses of relaxin appear to increase elastolytic activity in stress urinary incontinence cells by decreasing the expression of alpha-1 antitrypsin in proliferative phase cells or increasing the total elastase activity in secretory phase cells. Fibroblast total elastase activity was inhibited by increasing concentrations of alpha-1 antitrypsin. CONCLUSION: Elastase activity appears to be increased in relaxin-stimulated stress urinary incontinence fibroblasts by either decreased inhibitor (alpha-1 antitrypsin) production or increased elastase activity.

Adult↗
PubMed2005

Surgical management of complex obstetric fistula in Eritrea.

OBJECTIVES: To evaluate the incidence of and demographic characteristics associated with obstetric fistula in Eritrea. To determine the outcomes of surgical repair of complex fistula in Eritrea by a visiting surgical team. METHODS: A surgical team comprising expert gynecologic surgeons traveled to Eritrea in September 2004. We evaluated 50 patients with genitourinary fistula and performed surgical repairs of these fistulas on 37 women via both vaginal and abdominal approaches. Demographic and basic medical data were obtained at the time of evaluation, and follow-up questionnaires were completed at 4 weeks postoperative. RESULTS: The majority of the women had fistulas related to obstructed labor at their first pregnancy unattended by any healthcare professional. The average duration of labor was 3 days, and more than half had resulted in stillbirths. The rate of successful repair in women with primary vesicovaginal fistulas (VVF) was 63%, and that in women with recurrent vesicovaginal fistulas was 61%. Two women required urinary diversion procedures because of the severity of the damage to the genital tract. Urethral reconstruction in women with urethrovaginal fistulas (UVFs) was successfully accomplished in 77% of patients. The rate of successful repair of rectovaginal fistulas (RVFs) was 87%. CONCLUSIONS: We have demonstrated that a team of specialized surgeons can successfully accomplish surgical procedures and repairs of very complex urinary tract fistulas in a very short mission to a resource-poor nation.

Adult↗
PubMed2004

Elastolytic activity in women with stress urinary incontinence and pelvic organ prolapse.

AIMS: Weakening of pelvic supportive tissues is thought to be a contributing etiology in female pelvic floor disorders such as stress urinary incontinence and/or pelvic organ prolapse (SUI/POP). Since elastin modulates the mechanical properties of supportive tissues, we examined elastase activity in vaginal tissue from women with pelvic floor dysfunction compared to asymptomatic controls, by comparing overall elastase activity, human neutrophil elastase, cathepsin K, and alpha-1 antitrypsin (a serine protease inhibitor) mRNA and protein levels. METHODS: Full-thickness peri-urethral vaginal wall tissues were collected from age and menstrual-phase matched SUI/POP and control women at the time of pelvic surgery. Elastolytic activity in the homogenized tissue was determined by the generation of amino groups from succinylated elastin. To quantify mRNA levels of each protein, quantitative competitive-PCR and confirmatory Western blot analyses were performed on the samples for human neutrophil elastase, cathepsin K, and alpha-1 antitrypsin. RESULTS: The mean elastolytic activity in vaginal tissues from the SUI/POP group was similar to that in the control group. With respect to the proteolytic enzymes, neither human neutrophil elastase nor cathepsin K differed between the two groups. However, alpha-1 antitrypsin mRNA and protein levels were significantly decreased in tissues from affected women. CONCLUSIONS: A significant decrease in alpha-1 antitrypsin expression was seen in tissues from women with SUI/POP compared to controls. This data suggest that altered elastin metabolism may contribute to the connective tissue alterations observed in pelvic floor dysfunction. Future investigations are warranted to help define the role of elastin turnover in pelvic floor dysfunction.

Adult↗
PubMed2004

Estrogen bioassay of ginseng extract and ArginMax, a nutritional supplement for the enhancement of female sexual function.

PURPOSE: To determine whether ArginMax (The Daily Wellness Co., Sunnyvale, CA) or the Panax ginseng extract it contains has any estrogenic activity. ArginMax for Women, a nutritional supplement for optimization of sexual health, contains L-arginine, ginseng, ginkgo, damiana, multivitamins, and minerals. METHODS: A human endometrial adenocarcinoma cell line, Ishikawa, which contains an alkaline phosphatase (AP) enzyme sensitive to estrogen stimulation, was used in a bioassay to determine whether Panax ginseng or ArginMax contained estrogenic components. RESULTS: Neither ArginMax nor Panax ginseng stimulated AP at any of the concentrations tested. CONCLUSIONS: No estrogenic activity was evident in the sample of Panax ginseng extract tested or in a sample of ArginMax containing this extract in combination with other ingredients.

Adenocarcinoma↗
PubMed2004

Career advancement for women faculty in a U.S. school of medicine: perceived needs.

PURPOSE: The percentage of women faculty at the professor level has remained at approximately 11%. The medical community could benefit from knowing what is required to attract, retain, and advance women in the academy. METHOD: The Committee on Women in Medicine and Science at Stanford University School of Medicine was charged with improving career success and well-being of women faculty. In 2001-02, a survey instrument including both needs and perceived school climate was sent to 309 women faculty. Responses were analyzed using one-way analyses of variance with Tukey follow-up tests. RESULTS: A total of 163 (53%) faculty responded. The highest ranked needs were a flexible work environment without negative consequences for women with young children (mean = 4.37 on a five-point scale); a three-month sabbatical from clinical and administrative duties (mean = 4.15); departmental mentoring for academic career development (mean = 4.13); and school/departmental administrative secretarial support for grant and manuscript preparation (mean = 4.11). Climate data obtained in 2002, compared to data from similar surveys in 1994 and 1995, showed a nonsignificant decrease in mean ratings for sexual harassment, gender discrimination, and gender insensitivity in the intervening years. Mean ratings for positive climate and cohesion increased between 1994 and 1995 but remained stable from 1995 to 2002. Results of the survey were presented to the dean, faculty, and staff of the medical school. CONCLUSION: Women faculty members were able to clearly indicate specific interventions that would improve their career success and sense of well-being. Since administrators are committed to serious consideration of these recommendations, this was a key step in advancing women's careers in academic medicine at Stanford.

Academic Medical Centers↗
PubMed2004

Avoiding menstruation: a review of health and lifestyle issues.

This article (1) reviews the decades-long history of short-term dosing regimens delaying the onset of expected, spontaneous menses or withdrawal bleeding in oral contraceptive users up to 20 days; (2) outlines treatment schedules that suppress menstrual bleeding for several months; and (3) evaluates the recently approved extended dosing regimen of 3 months' duration. For single-term postponement of normal menses, estrogen-progestogen combinations can be employed, starting about 7 days after ovulation. Oral contraceptive users can skip the 7-day pill-free period and continue with the active pills in the next package. The main focus of this review is the development of extended dosing schedules that result in cycles lasting 7 weeks up to several months and reduce the number of periods of bleeding and menstrual discomfort. Recently a dosing schedule was introduced into clinical use consisting of ethinyl estradiol, 30 microg, plus levonorgestrel, 150 microg/d, for 84 days, followed by 7 days of placebo. The pregnancy rate was < 1% for compliant women and 1.5% for all participants. A monophasic 21 + 7-day combination using the same daily doses had slightly higher pregnancy rates. The discontinuation rate for unscheduled bleeding and spotting was higher with extended dosing than with the conventional, 21 + 7 schedule.

Contraceptives, Oral, Combined↗
PubMed2004

A nutritional supplement for improving fertility in women: a pilot study.

OBJECTIVE: To determine the impact of nutritional supplementation on optimization of reproductive health in women. STUDY DESIGN: A double-blind, placebo-controlled pilot study was initiated to determine the effects of FertilityBlend (Daily Wellness Co., Sunnyvale, California), a proprietary nutritional supplement containing chasteberry and green tea extracts, L-arginine, vitamins (including folate) and minerals. Changes in progesterone level, basal body temperature, menstrual cycle, pregnancy rate and side effects were monitored. RESULTS: Thirty women aged 24-46 years who had tried unsuccessfully to conceive for 6-36 months completed the study. After 3 months, the supplement group (n = 15) demonstrated a trend toward an increase in mean midluteal phase progesterone level (from 8.2 to 12.8 ng/mL, P = .08) and a significant increase in the average number of days in the cycle with basal temperatures >37 degrees C during the luteal phase (6.8-9.7 days, P = .04). The placebo group (n = 15) did not show any notable changes after treatment in any of the parameters studied. After 5 months, 5 of the 15 women in the supplement group were pregnant (33%), and none of the 15 women in the placebo group were (P <.01). No significant side effects were noted. CONCLUSION: Nutritional supplementation may provide an attractive alternative or complement to conventional fertility therapy.

Adult↗
PubMed2003

Regulation of embryonic implantation.

The preimplantation embryo produces several factors during its development to signal its presence to the maternal organism. This paper will focus on the role of two distinctive cytokine and growth factor systems (interleukin-1 (IL-1) system and the vascular endothelial growth factor (VEGF) system) during early embryonic development and implantation. IL-1 receptor is expressed in the endometrium of various species and antagonising the biological effects of IL-1 leads to implantation failure in mice. We could show that this is due to an endometrial, not an embryonic effect. Furthermore, we could detect the expression of all components of the IL-1 system in preimplantation embryos from mice and humans. We could show a possible influence of IL-1 on other systems involved in embryonic implantation, including invasion (MMPs/TIMPs) and angiogenesis (VEGF), therefore suggesting a role of this cytokine family during early embryonic development. Immediately after contact to the endometrium, the embryo must induce angiogenesis to ensure its survival, VEGF is a potent angiogenetic growth factor. We have shown a cyclic regulation of the soluble VEGF-receptor, sflt, in human endometrium and have detected the expression of the transmembraneous VEGF-receptors, Flt-1 and kinase insert domain containing receptor (KDR) throughout the menstrual cycle. Furthermore, we have shown that the VEGF gene is one of the earliest genes activated during human preimplantation embryo development, giving rise to the assumption that VEGF is crucial for embryonic development.

Animals↗
PubMed2003

Interleukin-1beta regulation of gonadotropin-releasing hormone messenger ribonucleic acid in cultured human endometrial stromal cells.

OBJECTIVE: To investigate the role of interleukin-1beta (IL-1beta) in regulating GnRH mRNA expression in cultured human endometrial stromal cells using a modified semiquantitative competitive reverse transcription and polymerase chain reaction (PCR). DESIGN: A controlled study. SETTING: Clinical and academic research setting in a university medical center. PATIENT(S): Women undergoing hysterectomy for nonmalignant indications. INTERVENTION(S): Confluent stromal cell cultures treated with steroid hormones were stimulated with IL-1beta and attenuated by anti-IL-1beta antibody or IL-1 receptor antagonist. MAIN OUTCOME MEASURE(S): The human endometrial stromal cell expression of GnRH and its receptor were determined by PCR. Interleukin-1beta-mediated regulation of stromal cell GnRH mRNA expression was determined by quantitative competitive PCR. RESULT(S): The GnRH and GnRH receptor mRNA expression were amplified in cultured stromal cells by PCR and two rounds of nested PCR, respectively. Treatment with IL-1beta stimulated stromal cell GnRH mRNA expression at concentrations of IL-1beta above 10 IU/mL. Recombinant IL-1 receptor antagonist and anti-IL-1beta antibody attenuated the increase of gene expression of GnRH initiated by IL-1beta. CONCLUSION(S): These results provide indirect evidence that IL-1beta may play a crucial role at the level of embryo-maternal interaction by regulating stromal cell expression of GnRH and its receptor, both known to be important in mediating trophoblast invasion and placental hormone regulation.

Base Sequence↗
PubMed2003

Distinctive proliferative phase differences in gene expression in human myometrium and leiomyomata.

OBJECTIVE: To gain a comprehensive view of the gene expression and regulation involved in uterine leiomyomata and matched normal myometrium using oligonucleotide microarray-based hybridization analysis. DESIGN: Retrospective analyses of tissue obtained in a prospective randomized clinical study. SETTING: Academic institution. PATIENT(S): Seven patients with leiomyomata scheduled for surgery during the proliferative phase. INTERVENTIONS(S): Seven paired samples of leiomyomata and adjacent myometrium were obtained from patients undergoing hysterectomy. MAIN OUTCOME MEASURE(S): The total RNA extracted from leiomyomata and myometrium was used for gene expression profiling of 6800 human genes using high-density oligonucleotide microarrays. In addition, reverse transcriptase-semiquantitative polymerase chain reaction and immunohistochemistry were used to validate tumor-specific gene expression. RESULT(S): A comparison of expression patterns in each paired sample revealed 68 genes significantly up- or down-regulated in each paired tissue sample, of which 23 genes showed increased expression and 45 showed decreased expression in leiomyomata compared with normal myometrium. Cluster analysis supported the relevance of these candidate genes for distinguishing between normal myometrium and leiomyomata biologic activity. CONCLUSION(S): Expression profiling of uterine leiomyomata using high-density oligonucleotide microarrays yields signature patterns that reflect the distinctive differences between normal human myometrium and leiomyomata during the proliferative phase. These observations suggest that a number of genes are involved in the tumorigenesis of leiomyomata.

Down-Regulation↗