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Characterization of epidermal growth factor receptor in human endometrial cells in culture
Citations from the literature/International
Journal of Gynecology & Obstetrics 49 (199s) 87-97
difficulties with defecation, although the number of patients studied was small. Conclusions: Sacrocolpopexy is a successful operation for the correction of prolapse. Complications include the development of genuine stressincontinence, detrusor instability, voiding difftculty, and mesh infection,
MENORRHAGIA Evaluation of surgical options in menorrhagia Lalonde A. CAN
BR J OBSTET GYNAECOL SUPPL. 1994 101/l I (8-14) Total abdominal and vaginal hysterectomy has been the standard treatment for intractable and unmanageable menorrhagia for many years. However, in the last few years, hysteroscopictreatment of menorrhagia through surgical resection and/or ablation of the endometrial cavity has gained success.Over 625 000 hysterectomies are performed each year in the USA alone. More than 30% are done for menorrhagia as a primary diagnosis. Three times more hysterectomies are performed using the abdominal route than the vaginal approach. Morbidity is higher using the abdominal route alone. Complications of abdominal and vaginal hysterectomies will be briefly discussed.Menorrhagia caused by submucous fibroids can be treated hysteroscopically. A review of methods, complications, advantages and long-term follow up will be presented. Laparoscopically-assisted vaginal hysterectomy (LAVH) is the latest surgical option offered for menorrhagia. The advent of a hysteroscopic approach to the treatment of menorrhagia and careful analysis of the advantages and disadvantages of LAVH will be discussed. Assesment of medical treatments for menorrhagia Shaw R.W. GBR
BR J OBSTET GYNAECOL SUPPL. 1994 101/l I (15-18) Although usually not life-threatening, dysfunctional uterine bleeding (DUB) can causediscomfort and disruption to life for many women. It has been poorly researchedin the past, primarily becauseof difficulties in trying to accurately measureblood loss and response to treatment. There are several different therapies currently available but, for many, actual evidence of their efficacy is lacking from scientific data. Progestogensare the most frequently prescribed drugs for the treatment of DUB. Data support their use in anovulatory women but a number of comparative trials have shown that an overall reduction in blood loss of only 20% is achieved in ovulatory women. Their use,therefore, must be questioned as the first line of treatment. Combined oral contraceptives were at one time popular but whether the low-dose, current generation pills are equally effective awaits appropriate trials. Prostaglandin synthetase inhibitors can be useful, with up to a third of women with menorrhagia benefiting from a reduction of between 25% and 35% in blood loss. A proportionally greater reduction is seen in women with more excessivebleeding. Antilibrinolytic drugs have been shown to reduce menstrual blood loss in DUB by
95
50%and would be useful in women in whom estrogensare contraindicated. Gonadotrophin-releasing hormone analogues are highly effective becauseof their ability to induce amenorrhea, but long-term use is contraindicated because of their hypoestrogeniceffects. One other effective therapy for menorrhagia has been danazol. At a dosage of 200 mg daily, danazol has been shown to be highly effective in several open and randomized comparative trials, with a consistent reduction of blood loss of up to 75% being achieved with maintenance of a regular menstrual cycle. Reducing the dose to 100 mg daily often results in cycle irregularities, whereas increasing the dose to 400 mg daily induces amenorrhea. A daily dose of 200 mg of danazol is well tolerated and should be considered as a first-line option in DUB presenting as menorrhagia requiring medical management. Endometrial ablation in the treatment of menorrhagia Erian J. GBR
BR J OBSTET GYNAECOL SUPPL. 1994 101/l I (19-22) A prospective 5-year multicenter study, involving three UK gynecology centres with a special interest in endoscopic laser surgery, was set up to determine the safety, acceptability, clinical effectiveness and complications of neodymium yttriumaluminium-garnet laser ablation of the endometrium in the treatment of menorrhagia. A total of 2342 women with disabling menorrhagia that was unresponsive to medical therapy were involved. The main outcome measureswere: preoperative endometrial preparation, duration of laser ablation, intra- and postoperative complications, amenorrhea rate, oligomenorrhea rate, and the women’s subjective assessmentof treatment. No major complications occurred in the 2342 treatments, Nine (0.4%) casesof transient fluid overload, II (0.5%) of infection and live (0.2%) of uterine perforation occurred. None of the women required a laparotomy. The mean duration of the laser ablation was 24 min. The post-surgery amenorrhea rate was higher in women pretreated with danazol. Of the 1866women followed up for at least I year after treatment, 1043 (56%) developed complete amenorrhea, 701 (38%) reported continuing but satisfactorily reduced menstruation, and 122(7%) patients failed to improve with the first treatment (57 of these 122 women responded to a second laser ablation). Overall, 1744 (93%) had a satisfactory responseto laser ablation and only 33 (1.8%) required subsequent hysterectomy. In conclusion, this study showed that hysteroscopic endometrial laser ablation is an acceptable alternative to hysterectomy for the treatment of menorrhagia.
EXPERIMENTAL
GYNECOLOGY
Characterization of epidermal growth factor receptor in human endometrial cells in culture Watson H.; Franks S.; Bonney R.C. GBR
J REPROD FERTIL 1994 lOl/2 (415-420) The aim of the study was to determine the binding characteristics of the epidermal growth factor (EGF) receptor
96
Citations from the literature /International
Journal of Gynecology & Obstetrics 49 (1995) 87-97
in isolated human endometrial glands and stromal cells in culture. Stromal cells and glands were obtained from endometrial tissue by collagenase dispersion followed by sieve filtration. They were plated into 24-well multiwell plates in Ham’s FIO medium supplemented with 5% fetal calf serum and used at 70-80% confluence. Scatchard analysis revealed a single class of high-affinity binding sites in both cell types with apparent dissociation constants of 1.17 0.6 (n = 15) and 1.200.3 (n = 8) nmol I-I for stromal cells and glands, respectively. The concentration of receptors was higher in stromal cells than in glands, 719 377(n = 16)and 310 177(n = 8) fmol mg-1 protein, respectively. Epidennal growth factor labelled with “sI was displaced from the receptor by EGF and transforming growth factor, but not insulin, insulin-like growth factor, fibroblast growth factor, or platelet-derived growth factor. Binding was shown to be dependent on time and temperature. Downregulation of the receptor was demonstrated by preincubating cells with 5 nmol EGF l-l, which reduced receptor concentrations by 75%. 12-OTetradecanoylphorbol-13-acetate decreasedthe affinity of the receptor for EGF changing the dissociation constant from I .8 to 3.9 nmol I- I. A suitable system for investigating the regulation of this receptor in human endometrium was established.
Improved detection of fetal ceils from maternal blood with polymerase chain reaction
characteristics and endothelium-dependent and endotheliumindependent relaxation were examined. STUDY DESIGN: By meansof a small vesselmyograph arteries of mean normalized internal diameter 353.22 * 13.14 pm were studied under isometric conditions. Contractile function was assessedwith a variety of agonists, including angiotensin II, endothelin-I. the thromboxane mimetic U46619, prostaglandin E,, and prostaglandin F (2o). The effect of physiologic and supraphysiologic PO2 on vascular function was also examined. Relaxation was assessedin response to known endotheliumdependent vasodilators, including acetylcholine, bradykinin, histamine, and A23187 and to sodium nitroprusside (endothelium independent). The effect of indomethacin and the nitric oxide synthase inhibitor, N(G)-nitro-L-arginine methyl ester on contractile function was also evaluated. RESULTS: Sensitivity to sodium nitroprusside was reduced by a high POz. U46619 was the most potent constrictor agonist studied. The response of precontracted arteries to known endotheliumdependent vasodilators was minimal, other than for histamine, which led to modest relaxation. The constrictor response to U46619was increasedin the presenceof N(G)-nitro-L-arginine methyl ester. CONCLUSIONS: Oxygen tension may be an important determinant of relaxation in small placental arteries. Receptor-mediated release of endothelium-derived relaxing factor is not a major mechanism in the fetoplacental circulation.
Adkison L.R.; Andrews R.H.; Vowel] N.L.; Koontz W.L. USA
AM J OBSTET GYNECOL 1994 170/3(952-955) OBJECTIVE: The objective was to test the reliability of new deoxyribonucleic acid primers that have previously been used very efficiently by this laboratory with amniolysate samples to amplify a 248 bp Y-specific, repeated sequencefrom maternal blood during pregnancy. STUDY DESIGN: Blood samples were obtained from 50 women during weeks I I and I6 of pregnancy and were analyzed for the presence of the Y chromosome-specificsequences.RESULTS: Y-specific fragments were identified in I9 of 24 (79.2%) women after one complete amplification. A second amplification of these samples negative for Y-specific fragments revealed three additional samplespositive for the Y chromosome-specific fragment. Only two male fetusesremained unidentified. Overall, 91.7% male fetusesand 96% of all fetuses(48/50) in these women were correctly identified. CONCLUSIONS: The primers described in this study provide an additional or alternative tool for the determination, by meansof the polymerase chain reaction, of Y chromosomebearing cells in maternal circulation.
Fuoctioaal characteristics of small placental arteries
McCarthy A.L.; Woolfson R.G.; Evans B.J.; Davies D.R.; Raju SK.; Poston L. GBR
AM J OBSTET GYNECOL 1994 170/3(945-951) OBJECTIVES: The aim of this study was to investigate characteristics of placental arteries capable of influencing vasomotor tone in the fetoplacental vascular bed. Contractile
C-myc ami tomor suppressor gene product expression in develop ing aml term human trophoblast
Roncalli M.; Bulfamante G.; Viale G.; Springall D.R.;Alfano R.; Comi A.; Maggioni M.; Polak J.M.; Coggi G. ITA
PLACENTA 1994 I5/4 (399-409) Proliferation and differentiation of villous trophoblast during placental development, from an early stage to full-term, were investigated in routinely fixed and processedtissues, by meansof the immunocytochemical localization of the cell cyclerelated proto-oncogene c-myc and the ~53 and retinoblastoma susceptibility (Rb) tumor-suppressorgene products. The proliferative activity of the trophoblast was determined using an antibody against proliferating cell nuclear antigen (PCNA) which stains all proliferating cells in paraffin-embedded tissues. Diffuse nuclear immunoreactivity for PCNA, c-myc and Rb geneproducts was a consistent finding in early cytotrophoblast: c-myc product expression was also detectable in both layers of mid-gestation trophoblast. Only scattered cytotrophoblastic nuclei of early gestational placenta displayed immunostaining for ~53 gene product. In full-term placenta c-myc expression was undetectable while Rb gene product and PCNA immunoreactivity declined markedly. These results indicate that the expressionof the above genesis spatio-temporally regulated during placental development. A potential involvement of the oncosuppressorgeneproducts ~53 and Rb in the control of trophoblastic proliferation and of c-myc in the control of both the proliferative and differentiation pathways of trophoblastic cells is suggested.
Journal of Gynecology & Obstetrics 49 (199s) 87-97
difficulties with defecation, although the number of patients studied was small. Conclusions: Sacrocolpopexy is a successful operation for the correction of prolapse. Complications include the development of genuine stressincontinence, detrusor instability, voiding difftculty, and mesh infection,
MENORRHAGIA Evaluation of surgical options in menorrhagia Lalonde A. CAN
BR J OBSTET GYNAECOL SUPPL. 1994 101/l I (8-14) Total abdominal and vaginal hysterectomy has been the standard treatment for intractable and unmanageable menorrhagia for many years. However, in the last few years, hysteroscopictreatment of menorrhagia through surgical resection and/or ablation of the endometrial cavity has gained success.Over 625 000 hysterectomies are performed each year in the USA alone. More than 30% are done for menorrhagia as a primary diagnosis. Three times more hysterectomies are performed using the abdominal route than the vaginal approach. Morbidity is higher using the abdominal route alone. Complications of abdominal and vaginal hysterectomies will be briefly discussed.Menorrhagia caused by submucous fibroids can be treated hysteroscopically. A review of methods, complications, advantages and long-term follow up will be presented. Laparoscopically-assisted vaginal hysterectomy (LAVH) is the latest surgical option offered for menorrhagia. The advent of a hysteroscopic approach to the treatment of menorrhagia and careful analysis of the advantages and disadvantages of LAVH will be discussed. Assesment of medical treatments for menorrhagia Shaw R.W. GBR
BR J OBSTET GYNAECOL SUPPL. 1994 101/l I (15-18) Although usually not life-threatening, dysfunctional uterine bleeding (DUB) can causediscomfort and disruption to life for many women. It has been poorly researchedin the past, primarily becauseof difficulties in trying to accurately measureblood loss and response to treatment. There are several different therapies currently available but, for many, actual evidence of their efficacy is lacking from scientific data. Progestogensare the most frequently prescribed drugs for the treatment of DUB. Data support their use in anovulatory women but a number of comparative trials have shown that an overall reduction in blood loss of only 20% is achieved in ovulatory women. Their use,therefore, must be questioned as the first line of treatment. Combined oral contraceptives were at one time popular but whether the low-dose, current generation pills are equally effective awaits appropriate trials. Prostaglandin synthetase inhibitors can be useful, with up to a third of women with menorrhagia benefiting from a reduction of between 25% and 35% in blood loss. A proportionally greater reduction is seen in women with more excessivebleeding. Antilibrinolytic drugs have been shown to reduce menstrual blood loss in DUB by
95
50%and would be useful in women in whom estrogensare contraindicated. Gonadotrophin-releasing hormone analogues are highly effective becauseof their ability to induce amenorrhea, but long-term use is contraindicated because of their hypoestrogeniceffects. One other effective therapy for menorrhagia has been danazol. At a dosage of 200 mg daily, danazol has been shown to be highly effective in several open and randomized comparative trials, with a consistent reduction of blood loss of up to 75% being achieved with maintenance of a regular menstrual cycle. Reducing the dose to 100 mg daily often results in cycle irregularities, whereas increasing the dose to 400 mg daily induces amenorrhea. A daily dose of 200 mg of danazol is well tolerated and should be considered as a first-line option in DUB presenting as menorrhagia requiring medical management. Endometrial ablation in the treatment of menorrhagia Erian J. GBR
BR J OBSTET GYNAECOL SUPPL. 1994 101/l I (19-22) A prospective 5-year multicenter study, involving three UK gynecology centres with a special interest in endoscopic laser surgery, was set up to determine the safety, acceptability, clinical effectiveness and complications of neodymium yttriumaluminium-garnet laser ablation of the endometrium in the treatment of menorrhagia. A total of 2342 women with disabling menorrhagia that was unresponsive to medical therapy were involved. The main outcome measureswere: preoperative endometrial preparation, duration of laser ablation, intra- and postoperative complications, amenorrhea rate, oligomenorrhea rate, and the women’s subjective assessmentof treatment. No major complications occurred in the 2342 treatments, Nine (0.4%) casesof transient fluid overload, II (0.5%) of infection and live (0.2%) of uterine perforation occurred. None of the women required a laparotomy. The mean duration of the laser ablation was 24 min. The post-surgery amenorrhea rate was higher in women pretreated with danazol. Of the 1866women followed up for at least I year after treatment, 1043 (56%) developed complete amenorrhea, 701 (38%) reported continuing but satisfactorily reduced menstruation, and 122(7%) patients failed to improve with the first treatment (57 of these 122 women responded to a second laser ablation). Overall, 1744 (93%) had a satisfactory responseto laser ablation and only 33 (1.8%) required subsequent hysterectomy. In conclusion, this study showed that hysteroscopic endometrial laser ablation is an acceptable alternative to hysterectomy for the treatment of menorrhagia.
EXPERIMENTAL
GYNECOLOGY
Characterization of epidermal growth factor receptor in human endometrial cells in culture Watson H.; Franks S.; Bonney R.C. GBR
J REPROD FERTIL 1994 lOl/2 (415-420) The aim of the study was to determine the binding characteristics of the epidermal growth factor (EGF) receptor
96
Citations from the literature /International
Journal of Gynecology & Obstetrics 49 (1995) 87-97
in isolated human endometrial glands and stromal cells in culture. Stromal cells and glands were obtained from endometrial tissue by collagenase dispersion followed by sieve filtration. They were plated into 24-well multiwell plates in Ham’s FIO medium supplemented with 5% fetal calf serum and used at 70-80% confluence. Scatchard analysis revealed a single class of high-affinity binding sites in both cell types with apparent dissociation constants of 1.17 0.6 (n = 15) and 1.200.3 (n = 8) nmol I-I for stromal cells and glands, respectively. The concentration of receptors was higher in stromal cells than in glands, 719 377(n = 16)and 310 177(n = 8) fmol mg-1 protein, respectively. Epidennal growth factor labelled with “sI was displaced from the receptor by EGF and transforming growth factor, but not insulin, insulin-like growth factor, fibroblast growth factor, or platelet-derived growth factor. Binding was shown to be dependent on time and temperature. Downregulation of the receptor was demonstrated by preincubating cells with 5 nmol EGF l-l, which reduced receptor concentrations by 75%. 12-OTetradecanoylphorbol-13-acetate decreasedthe affinity of the receptor for EGF changing the dissociation constant from I .8 to 3.9 nmol I- I. A suitable system for investigating the regulation of this receptor in human endometrium was established.
Improved detection of fetal ceils from maternal blood with polymerase chain reaction
characteristics and endothelium-dependent and endotheliumindependent relaxation were examined. STUDY DESIGN: By meansof a small vesselmyograph arteries of mean normalized internal diameter 353.22 * 13.14 pm were studied under isometric conditions. Contractile function was assessedwith a variety of agonists, including angiotensin II, endothelin-I. the thromboxane mimetic U46619, prostaglandin E,, and prostaglandin F (2o). The effect of physiologic and supraphysiologic PO2 on vascular function was also examined. Relaxation was assessedin response to known endotheliumdependent vasodilators, including acetylcholine, bradykinin, histamine, and A23187 and to sodium nitroprusside (endothelium independent). The effect of indomethacin and the nitric oxide synthase inhibitor, N(G)-nitro-L-arginine methyl ester on contractile function was also evaluated. RESULTS: Sensitivity to sodium nitroprusside was reduced by a high POz. U46619 was the most potent constrictor agonist studied. The response of precontracted arteries to known endotheliumdependent vasodilators was minimal, other than for histamine, which led to modest relaxation. The constrictor response to U46619was increasedin the presenceof N(G)-nitro-L-arginine methyl ester. CONCLUSIONS: Oxygen tension may be an important determinant of relaxation in small placental arteries. Receptor-mediated release of endothelium-derived relaxing factor is not a major mechanism in the fetoplacental circulation.
Adkison L.R.; Andrews R.H.; Vowel] N.L.; Koontz W.L. USA
AM J OBSTET GYNECOL 1994 170/3(952-955) OBJECTIVE: The objective was to test the reliability of new deoxyribonucleic acid primers that have previously been used very efficiently by this laboratory with amniolysate samples to amplify a 248 bp Y-specific, repeated sequencefrom maternal blood during pregnancy. STUDY DESIGN: Blood samples were obtained from 50 women during weeks I I and I6 of pregnancy and were analyzed for the presence of the Y chromosome-specificsequences.RESULTS: Y-specific fragments were identified in I9 of 24 (79.2%) women after one complete amplification. A second amplification of these samples negative for Y-specific fragments revealed three additional samplespositive for the Y chromosome-specific fragment. Only two male fetusesremained unidentified. Overall, 91.7% male fetusesand 96% of all fetuses(48/50) in these women were correctly identified. CONCLUSIONS: The primers described in this study provide an additional or alternative tool for the determination, by meansof the polymerase chain reaction, of Y chromosomebearing cells in maternal circulation.
Fuoctioaal characteristics of small placental arteries
McCarthy A.L.; Woolfson R.G.; Evans B.J.; Davies D.R.; Raju SK.; Poston L. GBR
AM J OBSTET GYNECOL 1994 170/3(945-951) OBJECTIVES: The aim of this study was to investigate characteristics of placental arteries capable of influencing vasomotor tone in the fetoplacental vascular bed. Contractile
C-myc ami tomor suppressor gene product expression in develop ing aml term human trophoblast
Roncalli M.; Bulfamante G.; Viale G.; Springall D.R.;Alfano R.; Comi A.; Maggioni M.; Polak J.M.; Coggi G. ITA
PLACENTA 1994 I5/4 (399-409) Proliferation and differentiation of villous trophoblast during placental development, from an early stage to full-term, were investigated in routinely fixed and processedtissues, by meansof the immunocytochemical localization of the cell cyclerelated proto-oncogene c-myc and the ~53 and retinoblastoma susceptibility (Rb) tumor-suppressorgene products. The proliferative activity of the trophoblast was determined using an antibody against proliferating cell nuclear antigen (PCNA) which stains all proliferating cells in paraffin-embedded tissues. Diffuse nuclear immunoreactivity for PCNA, c-myc and Rb geneproducts was a consistent finding in early cytotrophoblast: c-myc product expression was also detectable in both layers of mid-gestation trophoblast. Only scattered cytotrophoblastic nuclei of early gestational placenta displayed immunostaining for ~53 gene product. In full-term placenta c-myc expression was undetectable while Rb gene product and PCNA immunoreactivity declined markedly. These results indicate that the expressionof the above genesis spatio-temporally regulated during placental development. A potential involvement of the oncosuppressorgeneproducts ~53 and Rb in the control of trophoblastic proliferation and of c-myc in the control of both the proliferative and differentiation pathways of trophoblastic cells is suggested.