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Biomedical subjects

E J Thomas

Publications and source records attributed to E J Thomas.

At least 91 records · Page 5Linked to original sources

Pituitary and ovarian hormone levels in unexplained menorrhagia.

OBJECTIVE: To compare pituitary and ovarian hormone levels in women complaining of unexplained menorrhagia with objectively heavy menstrual blood loss (greater than 80 mL) to those with normal loss, and to determine whether there is any relation between menstrual volume and important cyclic endocrine events. METHODS: Over the course of 1 month, daily plasma LH, FSH, and estradiol (E2), and salivary progesterone concentrations were determined in 20 women with a measured menstrual loss exceeding 80 mL. These were compared with values in 22 women whose loss was less than 80 mL. For all participants, we calculated correlation coefficients for measured menstrual blood loss and the mean LH peak concentration, the mean FSH peak concentration, the mean of plasma E2 concentrations from day LH -3 to +1, and the mean of salivary progesterone concentrations from day LH +2 to +10. RESULTS: There were no significant differences in the plasma concentrations of LH, FSH, or E2 or in the salivary concentrations of progesterone between those with objectively heavy loss and those with normal loss. We did not find significant relations between the measured menstrual blood loss and the mean LH and FSH peak concentrations, the plasma E2 concentrations daily around ovulation, or the salivary progesterone concentrations during the luteal phase. CONCLUSION: There does not appear to be a relation between ovarian and pituitary hormones and menstrual loss.

Adolescent↗

The actions of estradiol and epidermal growth factor in endometrial and endometriotic stroma in vitro.

OBJECTIVES: To compare the effects of E2 and epidermal growth factor (EGF) on DNA and protein synthesis in endometrial and endometriotic stromal cell culture. DESIGN: Measurement of DNA and protein synthesis in cell cultures by trichloroacetic acid precipitation of radioactively labeled macromolecules. SETTING: Tissue samples collected in operating theaters were processed in the cell culture laboratory. PATIENTS: Samples of endometrium and endometriosis were collected synchronously at laparotomy. INTERVENTIONS: Stromal cells were isolated from tissue samples. Cultures of endometrial and endometriotic stromal cells were treated with 0.272 ng/mL (1 nmol/L) E2 or 10 ng/mL (1.6 nmol/L) EGF. MAIN OUTCOME MEASURES: 3H-Thymidine and 3H-leucine incorporation were used to assess DNA and protein synthesis respectively in cell cultures. RESULTS: There was no response to 0.272 ng/mL (1 nmol/L) E2 with respect to DNA or protein synthesis in either endometrial or endometriotic stromal cells, whereas 10 ng/mL (1.6 nmol/L) EGF significantly stimulated DNA and protein synthesis in both cell types. The effect of EGF was significantly greater in endometrial stroma than in endometriotic stroma. CONCLUSIONS: There are differences in the stimulation of 3H-thymidine incorporation by 10 ng/mL (1.6 nmol/L) EGF in endometrial and endometriotic stromal cells in vitro.

Cells, Cultured↗

Endometriosis.

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Endometriosis↗

Bradykinin stimulation of electrogenic ion transport in epithelial layers of cultured human endometrium.

Primary cultures of glandular endometrial epithelial cells grown on permeable supports formed monolayers with a high transepithelial electrical resistance [1096 +/- 83 omega.cm2 (n = 34)] and displayed electrogenic ion transport as demonstrated by an inward short circuit current (Isc; 20 +/- 2 microA/cm2). Bradykinin, 10(-8)-10(-6) M, added to either the basolateral or apical solutions enhanced the inward ISC. The concentration-response curves for bradykinin were bell-shaped in nature. The ISC response was more sensitive to apical addition of bradykinin and the maximum response was also greater with apical bradykinin. The increases in ISC were accompanied by two- to three-fold increases in transepithelial conductance. Apical addition of amiloride, 10(-4) M, reduced the unstimulated ISC by 80%. In the presence of amiloride, the response to both apical and basolateral bradykinin was reduced by > 50% in 8 out of 18 layers, and the mean response was reduced by approximately 25%. The data are consistent with a physiological role for bradykinin in the control of the intrauterine electrolyte environment, mediated in part by enhanced Na+ absorption.

Amiloride↗

Immunohistochemical localization of acidic and basic fibroblast growth factors in normal human endometrium and endometriosis and the detection of their mRNA by polymerase chain reaction.

Growth factors play a role in the cyclical growth and vascularization of normal endometrium. Abnormal endometrial proliferation and neovascularization may result in endometriosis. This study determines the presence and localization of acidic and basic fibroblast growth factors (aFGF and bFGF respectively) in endometrium of normal women, and in normal and ectopic endometrium of women with endometriosis. Endometrium was obtained at curettage or hysterectomy for benign disease, or laparoscopy for endometriosis. aFGF- and bFGF-immunoreactivity was detected at different phases of the menstrual cycle by immunohistochemistry using primary polyclonal rabbit antibodies. Expression of mRNA for aFGF and bFGF was determined in normal endometrium by nested reverse transcriptase polymerase chain reaction (RT-PCR). aFGF- and bFGF-immunoreactivity were both detected in endometrium from normal women, and in normal and ectopic endometrium of women with endometriosis. The pattern of staining with the two different FGFs was the same: immunoreactivity was predominantly confined to glandular epithelial cells and did not change throughout the menstrual cycle. Little or only light staining was seen in stromal cells and myometrium, and the pattern of staining did not differ between endometriotic and normal tissue. The presence of mRNA for aFGF and bFGF was demonstrated in normal endometrium. The detection of aFGF and bFGF mRNA in normal endometrium and aFGF- and bFGF-immunoreactivity in normal and endometriotic tissues suggests that these peptides may play a role in the proliferation and angiogenesis of normal and ectopic human endometrium.

Base Sequence↗

Ion transport by human endometrial epithelia in vitro.

Human glandular endometrial epithelial cells were cultured on porous tissue culture inserts to form tight, confluent layers. These layers generated time-dependent modifications in the ionic composition of both apical and basolateral solutions. Increases in sodium and chloride concentrations in the basolateral fluid were accompanied by reciprocal decreases in the concentrations of these ions in the apical fluid. The potassium concentration was increased in the apical, while decreased in the basolateral, solution. The total calcium concentration was slightly elevated in the apical, as compared with the basolateral fluid, while there were no alterations in pH. The endometrial layers demonstrated a significant transepithelial potential difference, and when this value was substituted in the Nernst equation a prediction of the passive distribution of ions across the cells was possible, indicating that none of the ions were in equilibrium. Addition of the sodium channel blocker amiloride to the medium bathing the cell layers reduced the modifications in ionic composition of apical and basolateral solutions. The data are consistent with other data indicating an amiloride-sensitive sodium-absorptive function for the endometrial epithelium. The ability of these primary cultures of endometrial epithelial cells to reduce the sodium while increasing the potassium concentration of the apical fluid is qualitatively in agreement with the low sodium and high potassium concentrations reported for human uterine fluid. The data suggest a role for the endometrial epithelium in generating and maintaining the distinctive ionic composition of the intra-uterine environment.

Adult↗

The impact of cigarette smoking on the plasma concentrations of gonadotrophins, ovarian steroids and androgens and upon the metabolism of oestrogens in the human female.

This study compares the plasma gonadotrophin, oestradiol, and androgen and salivary progesterone concentrations in a single menstrual cycle between 25 normal pre-menopausal women who smoke cigarettes and 21 who are non-smokers. The effect of smoking on luteinizing hormone (LH) pulsatility and the urinary excretion of oestrogens is also described. Cigarette smoking did not consistently suppress LH pulsatility. There was no significant difference in the length of either the follicular or luteal phases. There were no significant differences in the mean plasma oestradiol concentrations in the follicular phase in smokers compared to non-smokers. There were no significant differences in the mean salivary progesterone concentration in the luteal phase in smokers compared to non-smokers. There was no significant difference in plasma concentrations of testosterone, androstenedione and dehydroepiandrosterone sulphate. There was also no significant difference between the urinary concentrations of oestradiol, oestrone or oestriol. We have been unable to demonstrate a detrimental effect of cigarette smoking on any of the important endocrine characteristics of the menstrual cycle, and we conclude that these data suggest that the anti-oestrogenic effect of smoking does not work through alterations in pituitary or follicular endocrine function or in alterations in the metabolism of oestrogens.

Adult↗

Bombesin and gastrin-releasing peptide stimulate electrogenic ion transport in cultured human endometrial epithelial cell layers.

Electrogenic ion transport across human endometrial epithelial cells grown as polarized monolayers on permeable supports was measured as an inward short-circuit current (Isc; 16.2 +/- 1.1 microA/cm2). Bombesin (10(-7) M) and human gastrin-releasing peptide (GRP; 10(-7) M) caused transient enhancement of this Isc. These effects were largely restricted to the basolateral surface of the cells; responses to apical peptide were modest in comparison with those to basolateral peptide. GRP and other bombesin-related peptides may have a role in regulation of endometrial epithelial ion transport in vivo and thereby influence the intra-uterine environment.

Bombesin↗

Detection and characterization of the eae gene of Shiga-like toxin-producing Escherichia coli using polymerase chain reaction.

In this study, the polymerase chain reaction (PCR) was used in the detection of the attaching and effacing (eae) gene of Shiga-like toxin-producing Escherichia coli (SLT-EC). Oligonucleotide primers, complementary to the 5' portion of the eae gene of the enteropathogenic E. coli E2348/69 (O127:H6) and of SLT-EC CL8 and EDL933 (O157:H7), generated PCR products of the predicted sizes with DNA from the majority of human clinical SLT-EC strains tested from O serogroups 5, 26, 103, 111, 121, 128, 145, and 157; all SLT-EC strains of O serogroups 5, 26, and 111 from cattle; and a minority of porcine SLT-EC strains (one strain each from O serogroups 107 and 130 and one rough strain). Five HaeIII digestion profiles were obtained for PCR products generated by amplification of a 2.3-kb DNA fragment from the 5' end of eae. The HaeIII profiles for SLT-EC O serogroups, such as 26, 103, and 157, differed from each other but were consistent among strains within these O serogroups. Oligonucleotide primer pairs complementary to the 3' end of either the O127:H6 E. coli or the O157:H7 eae nucleotide sequence only amplified DNA from E. coli strains from a few of the SLT-EC O serogroups examined. One primer pair with homology to the 3' nucleotide sequence of eae from E. coli O157:H7 appeared to be relatively specific for this O serogroup by PCR. No PCR products were obtained in amplification experiments with the eae primers using DNA from human SLT-EC of O serogroups 38 (1 0f 1) and 91 (3 or 3), 15 of 15 SLT-EC strains from edema disease, or 29 of 29 non-SLT-EC strains from pigs and calves with diarrhea.

Bacterial Adhesion↗

Reverse transcription with nested polymerase chain reaction shows expression of basic fibroblast growth factor transcripts in human granulosa and cumulus cells from in vitro fertilisation patients.

Basic fibroblast growth factor (bFGF) is a potent angiogenic factor that has also been implicated in granulosa cell and oocyte maturation. We now report the expression of messenger ribonucleic acid (mRNA) encoding bFGF in human granulosa and cumulus cells obtained at oocyte recovery in in vitro fertilisation patients. It was necessary to use the sensitive technique of a nested polymerase chain reaction (PCR) after reverse transcription (RT) to detect transcripts. This finding in conjunction with a recent report showing the presence of transcripts for transforming growth factor beta (TGF beta) in the same type of cells by PCR indicates that mechanisms are in place for controlling extracellular proteolysis and cell differentiation.

Base Sequence↗

Nagging and other drinking control efforts of spouses of uncooperative alcohol abusers: assessment and modification.

This article presents a conception of spouse drinking control and an approach to assessment and modification to reduce the customary drinking control efforts of spouses of alcohol abusers unmotivated to enter treatment. Modification of the nonalcoholic spouse's customary drinking control is offered as an important early step in helping to prepare him or her to become a positive rehabilitative influence. Based on its use in unilateral family therapy with 68 spouses of uncooperative alcohol abusers, procedural guidelines, criteria for use, and two case examples from a crossover experimental dyad are described. Also presented are clinical results illustrating the success of the program, benefits and conditions relating to its use, and areas of possible application.

Alcohol Drinking↗

Characterization of human purified epithelial and stromal cells from endometrium and endometriosis in tissue culture.

OBJECTIVE: To initiate in vitro cultures of separate stromal and epithelial elements from endometriotic tissue and to compare the characteristics of these cells with those of cultured endometrial cells. DESIGN: The study involved testing the viability of a culture system for endometriotic tissue and examination of the phenotype of the cells. SETTING: Fresh tissue samples were collected from the operating theater and transferred to the tissue culture laboratory. PATIENTS, PARTICIPANTS: Twenty patients undergoing laparotomy for endometriosis and patients undergoing surgery for benign conditions were recruited. INTERVENTIONS: Endometrium and endometriotic tissue were separated, cultured in vitro, and labeled by indirect immunofluorescence with monoclonal antibodies against cytoskeletal components and epithelial mucins. MAIN OUTCOME MEASURES: Endometriotic cells have been maintained in vitro and found to resemble endometrial cells closely. RESULTS: With respect to the staining patterns for cytokeratins 18 and 19, vimentin, and three different epithelial mucins, cultured cells from both endometrium and endometriotic tissue had similar properties. Cytokeratins were located in epithelial cells, and vimentin was expressed in both stromal and epithelial cells. The antimucin antibodies all gave distinct patterns of intracellular staining of epithelial cells. CONCLUSIONS: Our results indicate a close similarity between cultured stromal and epithelial cells from endometrium and endometriotic deposits. Culture of these cell populations will permit study of their properties and interactions and may provide some insight into the cause of endometriosis.

Antigens, Neoplasm↗

The expression of retinoic acid receptors in cultured human endometrial stromal cells and effects of retinoic acid.

Patterns of expression of retinoic acid receptors (RAR) in cultures of human endometrial stromal cells are described. Transcripts for all three classes of RAR were expressed in these cells but RAR-beta was expressed at a low level by comparison with RAR-alpha and RAR-gamma. The abundance of RAR-beta transcripts was elevated by treating the cells with retinoic acid, but there was no effect on the level of expression of RAR-alpha and RAR-gamma. The induction of RAR-beta by retinoic acid was detectable within 4 h and at low concentrations of retinoic acid (10(-10) M). Adenosine 3':5'-cyclic monophosphate (cAMP) analogues and forskolin, an adenylate cyclase activator, had no effect on the retinoic acid-mediated induction of RAR-beta, contrary to recent observations on embryonal carcinoma cells. However, the phosphodiesterase inhibitor, 3-isobutyl-1-methyl-xanthine (IBMX), forskolin and 8-bromo-cAMP depressed basal levels of RAR-beta expression. These data suggest that endometrial stromal cells may be a target tissue of retinoic acid in vivo, and imply a role for retinoic acid in the cyclical differentiation of human endometrium.

Blotting, Northern↗

Ovarian steroid receptor expression in endometriosis and in two potential parent epithelia: endometrium and peritoneal mesothelium.

Endometriosis is an oestrogen dependent condition and it is expected that the tissue of origin of endometriosis will express receptors for the ovarian steroids. Two epithelia, endometrium and peritoneal mesothelium, are the potential parent epithelium. Oestrogen and progesterone receptor expression has been studied immunohistochemically in (i) timed endometrial biopsies from 25 normal subjects and 27 patients with endometriosis, (ii) 25 endometriotic biopsies and (iii) 42 peritoneal biopsies. Endometrium but not peritoneal mesothelium expresses both oestrogen and progesterone receptors. No difference in the intensity of staining between endometria of normal subjects compared with the endometria of patients with endometriosis was noted. In paired endometrial and endometriotic biopsies, the intensity of staining for the oestrogen receptor in stromal cells and for the progesterone receptor in both glandular and stromal cells was less in the endometriotic biopsies. These data provide circumstantial evidence for an endometrial origin for endometriosis although quantitative differences exist in receptor expression between endometrium and endometriosis.

Endometriosis↗

A randomized double blind placebo controlled trial of the effects on bone metabolism of the combination of nafarelin acetate and norethisterone.

OBJECTIVE: We observed the effects on bone metabolism of the addition of different doses of oral norethisterone during treatment with the GnRH agonist nafarelin (Synarel, Syntex). PATIENTS: Ninety-four women with a subjective complaint of heavy menstrual blood loss or objective evidence of endometriosis received intra-nasal nafarelin 400 micrograms daily for 6 months and also received, in a randomized, double blind manner, either 0.7 mg (n = 24), 1.4 mg (n = 23) or 2.45 mg (n = 23) of oral norethisterone or placebo (n = 24) daily. Follow-up was continued for a further 6 months after treatment. RESULTS: Thirty-one patients (33%) left the study prematurely and three patients were non-compliant with the study drug. By 6 months significant increases in urinary calcium/creatinine ratio were seen, compared to baseline, in the nafareline and placebo (P = 0.001, n = 14), 0.7 mg (P = 0.04, n = 13) and 1.4 mg norethisterone groups (P = 0.009, n = 17) but not in the nafarelin or 2.45 mg norethisterone groups (P = 0.72, n = 16). Densitometry of the spine, however, showed decreases at 6 months in all groups: 6.14% (P = 0.0004, n = 11), 5.46% n = 0.0006, n = 13), 3.93% (P = 0.008, n = 14) and 4.04% (P = 0.004, n = 16) for the groups receiving nafarelin and placebo, nafarelin and norethisterone 0.7, 1.4 and 2.45 mg respectively. Six months after stopping nafarelin, with or without norethisterone, bone mass was not different from baseline. CONCLUSION: The concomitant daily use of up to 2.45 mg of norethisterone does not eliminate the bone demineralization seen during GnRH agonist therapy with nafarelin in premenopausal women.

Administration, Oral↗