Biomedical subjects

N Chegini

Publications and source records attributed to N Chegini.

At least 19 recordsLinked to original sources

PubMed2006

Endometrial expression of epithelial neutrophil-activating peptide-78 during the menstrual cycle or in progestin-only contraceptive users with breakthrough bleeding and the influence of doxycycline therapy.

BACKGROUND: Endometrial breakthrough bleeding is characterized by an inflammatory reaction and increased production of proinflammatory mediators, one of which may be epithelial neutrophil-activating peptide-78 (ENA-78), a chemokine with neutrophil-activating properties. METHODS AND RESULTS: We therefore investigated the endometrial expression of ENA-78 in Norplant users as progestin-only contraceptive with various bleeding patterns (n=35) as compared with non-users with a normal menstrual cycle (n=55). The endometrial stromal cells (ESCs) were the major site of ENA-78 expression with the highest levels found during the secretory phase. The expression of ENA-78 was increased in Norplant users with irregular bleeding as compared with those with regular cycles and amenorrhoea. The levels of ENA-78 detected in uterine washes and sera after the use of Norplant for 3-6 months (n=25) increased compared with baseline (P < 0.05). These levels did not significantly change in Norplant users who received doxycycline (Dox) therapy (25 mg/twice daily for 6 months) when measured midway through or at the conclusion of study when compared with the baseline (n=25). Treatments with medroxyprogesterone acetate (MPA) and tumour necrosis factor-alpha (TNF-alpha) (25 ng/ml), but not 17beta-estradiol (E2) or E2 + MPA (10(-8) M), representing endometrium exposed to contraceptive and inflammatory conditions, respectively, increased the levels of ENA-78 production by ESCs, and this was reduced by co-treatments with Dox (25 microg/ml) (P < 0.05). CONCLUSIONS: The endometrial production of ENA-78 is altered in progestin-only contraceptive users experiencing breakthrough bleeding and is regulated by MPA and TNF-alpha in ESCs. Although Dox therapy did not alter uterine ENA-78 secretion, its suppression in ESCs suggests that Dox, acting site-specifically and through an anti-inflammatory mechanism, may influence the outcome of breakthrough bleeding in contraceptive users.

Adolescent↗

PubMed2002

Effects of GnRH analogues, 'add-back' steroid therapy, antiestrogen and antiprogestins on leiomyoma and myometrial smooth muscle cell growth and transforming growth factor-beta expression.

The objective of this study was to elucidate the biological significance of GnRH and antiprogestins and antiestrogen in leiomyoma and their interactions with ovarian steroid 'add-back' therapy. Leiomyoma and myometrial smooth muscle cells (LSMC and MSMC) were isolated and exposed to GnRH agonist (leuprolide acetate, LA), 17beta-estradiol (E2), medroxyprogesterone acetate (MPA), GnRH antagonist (Antide), estrogen antagonist, ICI182780 (Fulvestrant) and progesterone antagonists RU486 (Mifepristone) and ZK98299 (Onapristone) and combinations thereof. The rate of DNA synthesis, cell proliferation and transforming growth factor-beta (TGF-beta) expression were then determined. In both cell types, we found that in a dose-dependent manner, LA inhibited, whereas E2, MPA and the combination of E2 + MPA stimulated, the rate of DNA synthesis in these cells. Antide reversed the inhibitory effect of LA, while LA partly inhibited the stimulatory effect of the steroids. In addition, RU486, ICI182780 and ZK98299 at 0.1 micro mol/l or higher doses inhibited the rate of DNA synthesis and partly reversed the effects of E2 and/or MPA. We also found that LSMC expressed elevated levels of TGF-beta1 compared with MSMC. In both cell types, the effects of LA, E2, MPA, RU, ZK and ICI and combinations thereof on TGF-beta1 production were reflective of their effects on DNA synthesis. In line with this, TGF-beta1 was found to stimulate DNA synthesis and the E2-, TGF-beta1- or E2 + TGF-beta1-induced DNA synthesis was found to be inhibited by TGF-beta1 neutralizing antibodies and/or LA. In conclusion, the results provide further evidence that GnRH agonist- and RU486-induced leiomyoma regression is mediated in part through an interactive mechanism that results in altered cell growth and suppression of TGF-beta production.

Estradiol↗

PubMed2001

The expression and action of granulocyte macrophage-colony stimulating factor and its interaction with TGF-beta in endometrial carcinoma.

OBJECTIVE: Previous studies have demonstrated that normal human endometrium expresses granulocyte macrophage-colony stimulating factor (GM-CSF) and GM-CSF receptors. Because GM-CSF is administer to cancer patients following chemotherapy, GM-CSF may directly or through interaction with ovarian steroids and other cytokines alter the behavior of endometrial cancer. The aim of this study was to determine the expression of GM-CSF and receptors in endometrial carcinoma and its direct effect and interaction with transforming growth factor beta (TGF-beta) on Ishikawa cells, a human endometrial carcinoma cell line. METHODS: GM-CSF, GM-CSF receptors, TGF-beta1, and TGF-beta type II receptor expression were evaluated using quantitative reverse transcription polymerase chain reaction (Q-RT-PCR). The effect of GM-CSF on DNA synthesis, cell proliferation, expression of GM-CSF, TGF-beta1, and TGF-beta receptor, and their regulation by ovarian steroids was determined by the rate of [(3)H]thymidine incorporation, MTT assay, Q-RT-PCR, and ELISA, respectively. RESULTS: Endometrial carcinomas express significantly higher GM-CSF and GM-CSF alpha and beta receptor mRNA compared with normal postmenopausal endometrium. GM-CSF at various doses had no significant effect on the rate of [(3)H]thymidine incorporation or proliferation of Ishikawa cells, whereas TGF-beta1 inhibited [(3)H]thymidine incorporation. GM-CSF and TGF-beta1 regulate their own expression and the expression of TGF-beta type II receptor, which were both upregulated by 17beta-estradiol and medroxyprogesterone acetate treatment and reversed following cotreatment with their respective receptor antagonists. CONCLUSION: Endometrial carcinoma expresses an elevated level of GM-CSF and GM-CSF receptors. GM-CSF is not a mitogen for the endometrial cancer cell line; however, either alone or through interaction with TGF-beta1, it regulates its own expression and the expression of TGF-beta1 and TGF-beta type II receptor which inhabits endometrial cancer cells. This interaction may represent a regulatory feedback mechanism that could serve to suppress endometrial carcinoma growth.

Antineoplastic Agents, Hormonal↗

PubMed2001

Differential expression of integrin alpha v and beta 3 in serosal tissue of human intraperitoneal organs and adhesion.

OBJECTIVE: To assess the expression of integrin alpha v and beta 3 in the serosal tissue of intraperitoneal organs and adhesions in persons with and without adhesions. DESIGN: Prospective study. SETTING: Academic research centers. PATIENT(S): Fifty-seven patients undergoing abdominal or pelvic surgery. MAIN OUTCOME MEASURE(S): Integrin alpha v and beta 3 messenger RNA (mRNA) expression was evaluated by quantitative reverse transcription polymerase chain reaction. RESULT(S): The serosal tissue of the parietal peritoneum, uterus, fallopian tubes, ovary, and the large and small bowel, as well as peritoneal adhesions, skin, fascia, subcutaneous tissue, and omentum, expresses integrin alpha v and beta 3 mRNA. The level of alpha v and beta 3 mRNA expression varied among these tissues; expression of the former substance was highest in uterine serosa and lowest in the skin and small bowel, and expression of the latter substance was highest in the fallopian tubes and skin and lowest in the uterine serosa. Parietal peritoneum and adhesions express equal levels of integrin alpha v; however, integrin beta 3 expression was >100-fold lower in adhesions than in peritoneum. The level of integrin beta 3 expression in omentum, small and large bowels, and subcutaneous tissue was 100-fold to 10,000-fold lower than in other tissues. CONCLUSION(S): Serosal tissue of peritoneal organs and adhesions express variable levels of integrin alpha v and beta 3 mRNA. On the basis of such variation and the knowledge that tissue injury alters local integrin expression, integrins may play a key role in adhesion development, particularly in tissue with higher integrin expression.

Adult↗

PubMed2001

Matrix metalloproteinase (MMP-1) and tissue inhibitor of MMP in peritoneal fluids and sera and correlation with peritoneal adhesions.

OBJECTIVE: To assess the presence of matrix metalloproteinase (MMP-1) and tissue inhibitor of MMP (TIMP-1) in peritoneal fluid and serum of subjects with and without adhesions. DESIGN: Cross-sectional study. SETTING: Academic research centers. PATIENT(S): Sixty-three patients who underwent abdominal/pelvic surgery. INTERVENTION(S): MMP-1, TIMP-1, and MMP-1-TIMP-1 complex content. MAIN OUTCOME MEASURE(S): ELISA. RESULT(S): Peritoneal fluids (PF) and sera of subjects with and without peritoneal adhesions contain MMP-1, TIMP-1, and MMP-1-TIMP-1 complex at varying levels with 10- to 100-fold higher TIMP-1 than MMP-1. Compared with serum, PF contains a lower level of MMP-1 in subjects with mild adhesions and without adhesions, higher TIMP-1 in subjects with extensive adhesions, and lower MMP-1-TIMP-1 complex in subjects with moderate adhesions. However, the serum and PF content of MMP-1, TIMP-1, and MMP-1-TIMP-1 complex was not statistically different among subjects with or without adhesions, with the exception of TIMP-1 in PF of subjects with extensive adhesions. MMP1-TIMP-1 ratio indicates that a major portion of MMP-1 is in complex with TIMP-1. There was no age- or gender-dependent difference in MMP-1 and TIMP-1 content in serum or PF. CONCLUSION(S): Despite differences in MMP-1 and TIMP-1 levels in serum and PF of subjects with extensive and moderate adhesions, there is no correlation between MMP-1 and TIMP-1, with the exception of higher TIMP-1 in PF of subjects with extensive adhesions.

Abdomen↗

PubMed2001

Expression of matrix metalloproteinase (MMP-1) and tissue inhibitor of MMP in serosal tissue of intraperitoneal organs and adhesions.

OBJECTIVE: To compare expression of matrix metalloproteinase (MMP-1) and tissue inhibitor of MMP (TIMP-1) in serosal tissue of intraperitoneal organs and adhesions. DESIGN: Prospective and cross-sectional study. SETTING: Academic research centers. PATIENT(S): Patients undergoing abdominal or pelvic surgery. INTERVENTION(S): MMP-1 and TIMP-1 expression. MAIN OUTCOME MEASURE(S): Expression of messenger ribonucleic acid (mRNA) and protein was measured by using quantitative reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay. RESULT(S): Serosal tissue of intraperitoneal organs and adhesions express MMP-1 and TIMP-1 mRNA and protein at levels that are consistently varied with 10- to 10,000-fold and 2- to 10-fold higher TIMP, mRNA and protein, respectively. Parietal peritoneum, fallopian tubes and ovaries express higher MMP-1 mRNA levels compared with uterus and adhesions; the lowest expression is found in small and large bowels, subcutaneous tissue. and omentum. Expression of TIMP-1 mRNA was less variable; the highest level was found in the uterus and the lowest in subcutaneous tissue and small bowels. There was less variability in MMP-1 and TIMP-1 protein content than mRNA expression; ovaries and adhesions contained the highest MMP-1 and TIMP-1 levels, respectively, and peritoneum contained the lowest. The MMP-1 and TIMP-1 content and ratios further indicate limited MMP-1 proteolytic activity. Although tissues from premenopausal women express more MMP-1 and TIMP-1, expression did not differ by sex or age. CONCLUSION(S): Because MMP-1 and TIMP-1 expression varies consistently among the serosal tissues of peritoneal organs and adhesions, and because tissue injury alters their expression, site-specific variations in expression of these substances may predispose a particular organ to develop more adhesions.

Abdomen↗

PubMed2001

Granulocyte colony-stimulating factor in preterm and term pregnancy, parturition, and intra-amniotic infection.

OBJECTIVE: [corrected] To determine the sources of granulocyte colony-stimulating factor (G-CSF) in amniotic fluid and to examine its relation to labor and clinically diagnosed intra-amniotic infection. METHODS: We assessed G-CSF and G-CSF receptor expression in placentas (n = 50) from 5-40 weeks' gestation, and G-CSF concentrations were measured in amniotic fluid (n = 146), bronchoalveolar lavage fluid (n = 8), and paired maternal serum, cord blood, neonatal serum, and neonatal urine samples (n = 16). RESULTS: Immunohistochemical staining and messenger RNA analysis showed placental expression of G-CSF and G-CSF receptor throughout gestation. The number of decidual stromal cells expressing G-CSF receptor was significantly higher in women with intra-amniotic infection compared with women without infection (27 +/- 2 versus 18 +/- 3 cells per high power field, P =.02). Amniotic fluid concentrations of G-CSF were not significantly different in noninfected preterm compared with term samples (1708 +/- 1673 versus 1612 +/- 2100 pg/mL, P =.9). Labor was not associated with a significant increase in amniotic fluid G-CSF concentrations (1864 +/- 3151 versus 1612 +/- 2100 pg/mL, P =.77, term labor versus no labor; 3335 +/- 5364 versus 1708 +/- 1673 pg/mL, P =.09, preterm). Concentrations of G-CSF in maternal serum, amniotic fluid, bronchoalveolar lavage fluid, and neonatal urine were increased during intra-amniotic infection (all P <.05). CONCLUSION: Amniotic fluid G-CSF concentrations were similar in preterm and term pregnancies and were not significantly influenced by labor. Intra-amniotic infection was associated with an increased number of placental cells expressing the G-CSF receptor and higher concentrations of G-CSF in amniotic fluid, maternal serum, neonatal urine, and neonatal bronchoalveolar lavage samples.

Adult↗

PubMed2001

Macrophage secretory products and sperm zona pellucida binding.

OBJECTIVE: To determine if exposure of human gametes to macrophage secretory products reduces sperm binding to the zona pellucida, and to determine which cytokine(s) may be responsible for this effect. METHODS: A human macrophage cell line was cultured and either activated with lipopolysaccharide for 2 hours and then washed or left unactivated. Culture-conditioned media from activated or unactivated cells was used in hemizona assay. Hemizonae were incubated with sperm suspended in culture medium from either unactivated macrophages or activated macrophages, with the matching hemizona incubated with sperm suspended in control medium. Matching hemizonae were incubated with sperm suspended in unactivated macrophage medium paired with sperm suspended in activated macrophage culture medium. Conditioned medium from activated macrophages was found to have elevated levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta, and transforming growth factor-beta, therefore, gametes were also exposed to these cytokines followed by the hemizona assay. After each incubation, the number of sperm tightly bound to the outer surface of each hemizona was determined. RESULTS: Exposure of gametes to activated and unactivated macrophage culture-conditioned media significantly decreases sperm binding to the zona pellucida, with medium from activated macrophages inducing the greatest effect (P < .05). Exposure of sperm to TNF-alpha significantly impaired sperm binding (P < .05), whereas other cytokines tested had no effect. CONCLUSION: These results suggest that macrophage secretory products in the basal and activated state may be a factor in endometriosis-associated infertility through the interference of sperm binding to the zona pellucida, and that TNF-alpha is a key cytokine responsible for this effect.

Culture Media, Conditioned↗

PubMed2001

Overproduction of transforming growth factor-beta1 (TGF-beta1) is associated with adhesion formation and peritoneal fibrinolytic impairment.

BACKGROUND: Reduction in peritoneal fibrinolytic capacity and increased transforming growth factor-beta1 (TGF-beta1) production are associated with adhesion development. This study investigated the expression of TGF-beta1 in peritoneal tissue, and possible correlation with components of the fibrinolytic system locally in peritoneal tissue. MATERIALS AND METHODS: Peritoneal samples were taken from 22 patients at relaparotomy. Samples of adhesions were collected from 10 patients. The patients were categorized into different groups depending on the quantity and the quality of adhesions. TGF-beta1 and components of the fibrinolytic system in tissue extracts were assayed using enzyme-linked immunosorbent assays. RESULTS: The concentration of active TGF-beta1 in peritoneal samples from patients with extensive adhesions was double (P <.01) that of healthy subjects, but the total levels of TGF-beta1 were similar (P =.63). In adhesion tissue, both active (P <.003) and total (P <.008) TGF-beta1 concentrations were more than twice as high as unaffected peritoneum. There was a significant correlation between the concentration of plasminogen activator inhibitor type 1 in peritoneal samples with active TGF-beta1 (P <.03, r = 0.693) and adhesion tissue with total TGF-beta1 (P =.001, r = 0.872). The other components of the fibrinolytic system did not correlate significantly with TGF-beta1. CONCLUSIONS: These data indicate that an overexpression of TGF-beta1 is associated with adhesion formation, possibly through a mechanism involving local regulation of plasminogen activator inhibitor type 1.

Abdomen↗

PubMed2001

Differential expression of TGF-beta1 and TGF-beta3 in serosal tissues of human intraperitoneal organs and peritoneal adhesions.

Elevated local expression of transforming growth factor (TGF-beta) has been associated with increased incidence of peritoneal adhesion formation. In this study we determine whether differences in basal expression of TGF-beta in serosal tissue of peritoneal organs correlate with incidence of adhesion formation. Serosal tissue of parietal peritoneum, uterus, oviduct, ovary, omentum, large and small bowels as well as adhesions, skin, fascia, subcutaneous tissue, peritoneal fluid and serum were collected from 57 subjects with/without adhesions who were undergoing abdominal/pelvic surgery. To determine TGF-beta1 and TGF-beta3 mRNA and protein expression, total RNA and protein were isolated from these tissues and along with the fluids, subjected to quantitative RT-PCR and enzyme-linked immunosorbent assay (ELISA) respectively. Tissue sections were immunostained for TGF-beta1 and TGF-beta3 protein. We found that TGF-beta1 and TGF-beta3 mRNA and protein are expressed in these tissues and present in peritoneal fluids and serum, with considerable variations in level of their expression. Comparatively, there was more variation in TGF-beta1 than TGF-beta3 expression without age or gender relation. Adhesions express a significantly higher TGF-beta1 mRNA and have the highest TGF-beta1:TGF-beta3 ratio, with lowest concentrations and ratio detected in omentum, small and large bowels; in contrast uterus expresses higher TGF-beta3, with lowest concentrations detected in subcutaneous tissue and large bowels (P < 0.05). A similar trend was also observed for total (active + latent) TGF-beta1 protein expression, with low active TGF-beta1 that was not significantly different among the tissue extracts and fluids. However, the lowest active:total TGF-beta1 ratio was found in adhesions and ovary. In subjects with adhesions, the adhesions express significantly more TGF-beta1 compared to parietal peritoneum (P < 0.05). Immunoreactive TGF-beta1 and TGF-beta3 protein were present in various cell types in these tissues with intensity reflecting their mRNA and protein expression. In conclusion, we provided evidence that serosal tissue of various peritoneal organs and adhesions express TGF-beta1 and TGF-beta3. Since TGF-beta is expressed differently in these tissues and tissue injury often alters the expression of TGF-beta, we propose that tissues with a higher basal expression of TGF-beta may become predisposed to develop more adhesions compared to others.

Adult↗

PubMed2001

Adhesion development and the expression of endothelial nitric oxide synthase.

OBJECTIVE: This study was conducted to determine whether nitric oxide (NO), a potent vasodilator and inhibitor of thrombus formation, is involved in the formation and maintenance of adhesions. METHODS: Skin, subcutaneous tissues, peritoneum and adhesions were collected from surgical patients and total RNA was isolated. Quantitative reverse transcription polymerase chain reaction (QRT-PCR) was performed to quantitate endothelial nitric oxide synthase (eNOS) and beta-actin mRNA levels. RESULTS: eNOS mRNA levels for skin, subcutaneous tissue, peritoneum and adhesions were < or = 3.12 x 10(-4), < or = 3.12 x 10(-4), 6.24 x 10(-4) and 2.5 x 10(-3) attomoles/microl, respectively. Beta-actin mRNA levels for all tissues were between 1.25 x 10(-1) and 6.25 x 10(-2) attomoles/microl. CONCLUSION: eNOS mRNA can be identified in tissue adhesions, and may therefore play a role in adhesion formation and maintenance.

Actins↗

PubMed2000

Differential regulation of mesothelial cell fibrinolysis by transforming growth factor beta 1.

Inflammation and tissue trauma during the surgical procedure reduce the peritoneal fibrinolytic capacity. These conditions promote adhesion formation, and are associated with increased expression of transforming growth factor beta 1 (TGF-beta1). The objective of the present study was to investigate whether TGF-beta1 regulates the expression of fibrinolytic components in peritoneal mesothelial cells. Human peritoneal mesothelial cells (HPMC) were cultured and treated with various concentrations of human recombinant TGF-beta1 (0.1, 1.0 and 10 ng/mL) for 24 h. Levels of tissue- and urokinase plasminogen activator (t-PA and uPA), plasminogen activator inhibitor type-1 (PAI-1) and type-2 (PAI-2) mRNA and protein were assessed by quantitative reverse transcriptase polymerase chain reaction (Q-RT-PCR) and ELISA, respectively. HPMC expressed these components at the gene and protein level. TGF-beta1 downregulated, dose-dependently t-PA mRNA and protein to about 50% of control values (p = 0.0010), and doubled PAI-1 protein production (p = 0.0008) compared to untreated controls. Although uPA gene expression increased in cells exposed to TGF-beta1, the corresponding protein concentration in conditioned media did not. PAI-2 was not affected, either at the gene or protein level. In conclusion, the results indicate that fibrinolytic capacity of mesothelial cells is reduced by TGF-beta1, suggesting that peritoneal adhesion formation induced by TGF-beta1 may be mediated, in part, through reduction in fibrin degradation capacity at an early stage of peritoneal tissue repair.

Enzyme-Linked Immunosorbent Assay↗

PubMed2000

Transforming growth factor beta isoforms production by human peritoneal mesothelial cells after exposure to hypoxia.

PROBLEM: Although human mesothelial cells (HMC) line nearly the entire abdominal cavity, little is known about their role in adhesion formation. This study determines the effect of hypoxia and transforming growth factor (TGF)-beta1 on the ability of HMC to produce TGF-beta1-3, which have been implicated as mediators of the healing process. METHOD OF STUDY: HMC were cultured under normal and hypoxic conditions, and treated with and without TGF-beta1 for 24 hr. RNA from each group was subjected to multiplex reverse transcription-polymerase chain reaction to quantitate TGF-beta1-3 mRNA levels. RESULTS: Hypoxia resulted in 2- and 3.3-fold increase, while TGF-beta1 treatment resulted in 1.4- and 1.2-fold increase (normoxia) and 0- and 4.8-fold increase (hypoxia) in TGF-beta1 and TGF-beta2 mRNA levels, respectively. There was no detectable TGF-beta3 mRNA in HMC before or after treatments. CONCLUSION: TGF-beta1 treatment under hypoxia further extenuates endogenous TGF-beta2 but blocks TGF-beta1 production, thereby decreasing the TGF-beta1/TGF-beta2 ratio, which may result in the reduction of scarring and fibrosis.

Cell Adhesion↗

PubMed2000

Enhanced expression of Fas-associated proteins in decidual and trophoblastic tissues in pregnancy-induced hypertension.

PROBLEM: To determine if feto-placental tissues from gestations complicated by pregnancy-induced hypertension (PIH) have altered expression of Fas-associated proteins. METHOD OF STUDY: The expression of several Fas-related proteins was determined in fetal membranes, decidua, and placentas obtained from PIH-affected (n = 12, age range 32-36 weeks) and normal (n = 6, age range 37-41 weeks) gestations. Paraffin-embedded tissue sections were stained with specific monoclonal antibodies to Fas, Fas ligand (FasL), caspase-3, and bax. RESULTS: We observed greater expression of Fas and FasL in amnion and decidua from PIH-affected gestations than in normal controls. Intense staining was observed only in the perivascular endothelium (caspase-3) and in decidual cells (bax) from PIH gestations. CONCLUSION: Differential expression of Fas-related proteins in fetal membranes, decidua, and placentas from PIH-affected gestations is consistent with increased apoptosis, and suggests activation of the Fas/FasL pathway in a tissue-specific manner.

Caspase 3↗

PubMed1999

Expression of integrin messenger ribonucleic acid in human endometrium: a quantitative reverse transcription polymerase chain reaction study.

OBJECTIVE: To assess the expression of selected integrin subunit's messenger RNA (mRNA) in human endometrium during the menstrual cycle. DESIGN: A prospective comparative study. SETTING: Academic research environment. PATIENT(S): Premenopausal women with histologically normal endometrium who were undergoing hysterectomy. INTERVENTION(S): Endometrial tissues were collected. RESULT(S): Human endometrium expresses integrins alpha2, alpha3, alpha4, alpha5, alpha6.1, alpha6.2, alpha v, beta1, beta2, beta3, and beta5, as well as fibronectin mRNA. The levels of endometrial integrins mRNA expression varied significantly, with the lowest levels observed for alpha2, beta3, and fibronectin and the highest for alpha5, beta2, and beta5. The levels of integrins alpha2, alpha3, and alpha5 mRNA expression were significantly higher during the proliferative phase, whereas alpha4, alpha6.2, alpha v, beta1, beta2, beta3, beta5, and fibronectin were higher during the secretory phase of the menstrual cycle. The alpha6.1 mRNA was found to be equally expressed in the endometrium during the menstrual cycle, whereas the most dramatic changes occurred in alpha v and beta3 expression, compared with other integrin subunits. CONCLUSION(S): Human endometrium expresses mRNA for several integrins and fibronectin, with up-regulation of alpha4, alpha v, beta1, and beta3 during the secretory phase of the menstrual cycle, suggesting that their differential expression may be regulated in part by ovarian steroids.

Adult↗

PubMed1999

Localization of granulocyte-macrophage colony-stimulating factor in the bovine reproductive tract.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) can increase embryo development to the blastocyst stage in cattle. The objective of the present study was to determine whether GM-CSF is present in the reproductive tract. Using Western blotting, immunoreactive GM-CSF was detected in uterine flushings from cows at days 0, 7, and 14 of the estrous cycle and from cows at days 14-17 of pregnancy. Also, GM-CSF was localized immunohistochemically to endometrium and oviduct. Patterns of immunohistochemical localization and intensity of reaction product were similar for all days of the estrous cycle. While present in several cell types, immunoreactive product in the endometrium was greatest in epithelium (especially luminal epithelium). Immunoreactive GM-CSF was also localized to epithelium in ampullary and isthmic regions of the oviduct, with intensity greater in ampulla. Staining was observed for both ciliated and non-ciliated cells. In conclusion, the bovine oviduct and endometrium contain immunoreactive GM-CSF and this molecule is present in uterine secretions. Thus, this cytokine is a potentially important intracellular regulator of endometrial, oviductal and embryonic function during early pregnancy in the cow.

Animals↗

PubMed1999

Postoperative exposure to glove powders modulates production of peritoneal eicosanoids during peritoneal wound healing.

OBJECTIVE: To assess the effect of postsurgical exposure of peritoneal cavity to glove powders, Hydrocote, latex proteins, and lipopolysaccharide (LPS) on eicosanoid production in peritoneal fluid and cellular distribution of eicosanoid enzymes in peritoneal wound during healing. DESIGN: Randomised experimental study. SETTING: Institute for Wound Research, USA. ANIMALS: 360 mice randomised into six groups of 60 each. INTERVENTION: Abrasion of peritoneal cavity followed by instillation of 500 microl of sterile phosphate buffered saline (PBS) alone (Control) or containing 100 microg/ml of Biosorb, Keoflo, Hydrocote, 1 mg/ml of latex proteins, or 12.5 microg/ml of LPS. Mice were killed at 1, 2, 4, 7, 14 and 28 days, and the peritoneal washing obtained from each animal and concentration of eicosanoids measured. Tissue were immunostained for cyclooxygenases and 5-lipoxygenase and thromboxane A2 (TXA2) synthetase. RESULTS: Peritoneal fluid from uninjured controls contained 3.9 (0.8), 5.2 (0.3) and 0.2 (0.02) ng/ml of thromboxane B2 (TXB2), prostaglandin E2 (PGE2) and leukotriene B4 (LTB4), respectively. These increased significantly during the first week to 6.3 (0.3), 11.7 (0.8) and 2.6 (0.1) ng/ml, p<0.05, before returning to baseline by day 14. In all the treated groups the values were significantly higher than in controls (p<0.05). Immunoreactive cyclo-oxygenases, 5-lipoxygenase and TXA2 synthetase proteins were present in various cell types in uninjured skin and peritoneum, incisional and peritoneal wounds and adhesion tissues. Staining was more intense at the site of wounds and paralleled eicosanoid concentrations during healing. There was no difference between exposed and unexposed groups. CONCLUSION: The presence of glove powders, latex proteins and LPS in peritoneal cavity cause increased eicosanoid production and aggravate the normal inflammatory reaction to tissue injury. This may contribute to the inflammatory or immune reactions and development of adhesions caused by glove powders.

Animals↗

PubMed1999

Regulation of matrix metalloproteinases (MMPs) and their tissue inhibitors in human myometrial smooth muscle cells by TGF-beta1.

The objective of the present study was to determine whether transforming growth factor beta (TGF-beta) regulates the expression of matrix metalloproteinases (MMP) and the tissue inhibitor of MMP (TIMP) in myometrial smooth muscle cells. Using primary cultures of human myometrial smooth muscle cells we found that these cells express MMP-1, MMP-3, TIMP-1 and TIMP-2 mRNA and protein, with significantly higher values of TIMP than MMP. We also found that TGF-beta1 (1 ng/ml) increased the expression of TIMP-1 mRNA, while it reduced the expression of MMP-1 and MMP-3 mRNA, compared with untreated controls. In addition, TGF-beta1 slightly increased the production of TIMP-1, but not TIMP-2. Production of MMP-1 and MMP-3 was reduced by treatment with TGF-beta1, compared with the untreated control. A major portion of MMP-1 released into the culture-conditioned media was in complex with TIMP-1, and the levels of this complex were reduced by treatment with TGF-beta1. In conclusion, the data indicate that myometrial smooth muscle cells express MMP and TIMP mRNA and protein, and their expression is differentially regulated by TGF-beta1. Such a differential regulation of MMP and TIMP by TGF-beta may influence the rate of extracellular matrix (ECM) turnover following tissue injury, induced during myomectomy and Caesarean section, or in leiomyomas during growth.

Cells, Cultured↗