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Two TRH-responsive elements have been identified in the rat TSH beta gene by deletion/mutation analysis of the 5'-flanking region of the gene and transfection of TSH beta-luciferase constructs into the GH3 pituitary cell line. Biological responsiveness was confirmed by inserting synthetic oligonucleotides next to the heterologous viral thymidine kinase (tk) promoter in tk luciferase (tkLUC) constructs. Both DNA regions, termed TSH A (at -274 to -258 bp) and TSH C (-402 to -385 bp), have a high level of sequence similarity to binding sites for the POU domain pituitary transcription factor Pit-1/GHF-1. In transfection assays, the TSH A region had no basal enhancer activity, but did confer 3- to 6-fold TRH- and PMA-stimulated transcriptional responses to the tk promoter. The TSH C region conferred basal enhancer activity (3- to 10-fold above control tkLUC) as well as a 2- to 3-fold TRH or PMA response. Combinations of TSH A and TSH C elements conferred both enhancer activity and a TRH- or PMA-stimulated response, but more than two copies of the regions resulted in no further stimulatory effect. Both TSH beta gene regions bound to nuclear proteins from GH3 cells, as determined by gel retardation analysis. The TSH A region DNA formed three prominent DNA-protein complexes, ranging from slowly to rapidly migrating bands and with calculated affinities of 32, 0.5, and 208 nM, respectively. The TSH C region formed two major complexes, which corresponded on the basis of mobility to the most slowly and rapidly migrating complexes formed by TSH A, but with calculated affinities of 3.1 and 33 nM. TSH C also formed a rapidly migrating minor complex unique for this gene region. The more rapidly migrating complexes appeared to be specific to nuclear proteins from GH3 cells. Treatment of cells with TRH did not significantly alter the affinity of protein binding. Mutation of TSH A and TSH C DNA by T to G substitutions abolished the ability of the DNA to confer a TRH response and severely inhibited the ability of the DNA to bind to GH3 nuclear proteins. Thus, transcriptional regulation of the rat TSH beta gene by TRH is correlated with the ability of the two TRH-sensitive elements to bind nuclear proteins. The differences noted in basal enhancer activity or the degree of TRH responsiveness may be related to some unique proteins bound to each DNA or to the differences in affinity of binding of the proteins common to both elements.

Sixty-three women with clinical and urodynamic evidence of stress incontinence were evaluated before and after incontinence surgery for symptoms of depression, nervousness, tension, sleep disturbances, decreased appetite, somatic weakness and headaches. Women treated successfully with surgery demonstrated a statistically significant improvement in their subjective psychologic status (P less than .05). Unsuccessful treatment, however, was not associated with a significant change in or deterioration of their symptoms. All the symptoms were evaluated individually to ascertain the specific effects of treatment. Sleep disturbances were significantly improved with successful treatment and worsened with unsuccessful treatment (P less than .05). Tension was significantly improved with successful therapy (P less than .05) but was unchanged if surgery was unsuccessful. Depression became worse with subjectively unsuccessful surgery. Headaches and appetite were not affected by the therapeutic outcome. Therapy can be instrumental in affecting the psychologic status of women with stress incontinence. If the psychologic disability continues after therapy and/or treatment is unsuccessful, a referral for psychologic evaluation should be considered.

Previous work from this laboratory demonstrated that 17-beta estradiol (E2) can directly stimulate the transcription rate of the rat luteinizing hormone beta (LH beta) gene and that an upstream portion of the LH beta gene between -2.0 and -0.6 kilobases could confer an E2-stimulated response to a reporter gene in transient expression assays. To localize the LH beta estrogen response element (ERE) by biological function, portions of the 5'-flanking region of the LH beta gene or synthetic oligonucleotides were inserted in expression vectors next to the herpes simplex virus thymidine kinase promoter fused to the chloramphenicol acetyltransferase gene. Constructs were transfected into GH3 cells, and transfected cells were treated for 48 h with E2. E2 stimulation of activity (2-4-fold) occurred with constructs containing the 15-base pair palindromic sequence (GGACACCATCTGTCC), found at bases -1173 to -1159 relative to the transcriptional start site in the LH beta gene. A construct containing a synthetic oligonucleotide of this putative LH beta ERE was stimulated 1.7-3-fold by E2, while a construct containing two copies of the sequence was stimulated to a slightly higher level (2.5-4.0-fold). An oligonucleotide in which the palindrome was mutated failed to confer E2 stimulation, and mutation of the palindromic region within the upstream region of the LH beta gene also eliminated the E2 response. The anti-estrogen tamoxifen could not elicit a response, nor could dehydrotestosterone or dexamethasone; however, thyroid hormone treatment resulted in a 2-2.5-fold stimulation. The 15-base pair LH beta gene palindrome was found to bind estrogen receptor (ER) complex directly by gel retardation experiments. Labeled LH beta ERE DNA formed three complexes with proteins from immature rat uterine extract. Two of these were associated with ER complexes, as determined by the comigration of [3H] estradiol bound to ER with these complexes, and by the ability of anti-ER antibody to associate with these complexes. The affinity of the LH beta ERE for ER was calculated by Scatchard analysis to be 2.2-5.0 nM, an approximately 5-10-fold lower affinity than for the ERE in the vitellogenin A2 gene region. The mutated ERE, which had no biological activity, could not compete effectively for binding to ER. ER which was heat-transformed at 30 degrees C had a similar affinity (2-5 nM) for the ERE as ER occupied with E2 (2-4 nM), while ER occupied by estrone had a lower affinity (9 nM).(ABSTRACT TRUNCATED AT 400 WORDS)

Sixty-three women with genuine stress urinary incontinence were evaluated urodynamically both before and after modified Burch colposuspension to determine the association between pressure transmission ratio and leakage of urine during dynamic urethral pressure profile and the effect of surgery on pressure transmission ratio. Preoperatively, there was no difference in pressure transmission ratios between urethral pressure profiles associated with leakage of urine and those without urine leakage. Postoperatively, there were differences in pressure transmission ratios between leak and nonleak profiles in the sitting full and supine positions (P less than .001). We could not determine a threshold pressure transmission ratio that was more characteristically associated with leakage of urine. Finally, preoperative pressure transmission ratios were not associated with surgical success, but the higher the postoperative pressure transmission ratio, the more likely the surgery was to be successful (P = .023). In addition, the greater the difference between preoperative and postoperative pressure transmission ratios, the more likely was a cure (P = .011). The pressure transmission ratio may not be helpful in discriminating leak from nonleak dynamic urethral pressure profiles, and there may not be a threshold pressure transmission ratio below which patients characteristically leak urine. However, comparison of preoperative and postoperative pressure transmission ratios is helpful in determining the adequacy of surgical correction of the anatomical defect associated with genuine stress incontinence.

Thirty women with "atypical" squamous cells but not cervical intraepithelial neoplasia (CIN) on their Papanicolaou smears were tested for the presence of human papillomavirus (HPV) with the Southern blot hybridization technique. The Papanicolaou smears were reviewed for the presence of atypical squamous cells according to Patten's criteria. Comparison groups consisted of 30 patients with normal, 29 patients with CIN I and 24 patients with CIN II Papanicolaou smears. Ten of the 30 women (33%) with atypical cells on Papanicolaou smears were positive for HPV DNA as compared to 17% with normal and 59% with CIN I or II Papanicolaou smears. HPV 16 was present in 70-80% of the HPV DNA-positive patients in each Papanicolaou smear group. Concurrent CIN was also identified on colposcopically directed biopsies in 27% of the patients with squamous atypia. Patients with atypical squamous cells on Papanicolaou smears had coexisting HPV infection and CIN in about one-third of the cases. Colposcopy and further follow-up are recommended for such patients.

The carbon dioxide (CO 2) laser can be used to treat a variety of soft tissue disorders encountered by the urogynecologist. Urethral caruncles, red fleshy masses located at the urethral meatus, can be excised and vaporized using the CO 2 laser. Distal urethral diverticula, a common urologic disorder in women, can be marsupialized using the CO 2 laser. The epithelial lining of the diverticulum can be vaporized, thereby decreasing the risk of recurrence. Distal urethral stenosis may lead to symptoms associated with urethral syndrome and can be caused by a fibroelastic band of tissue. The CO 2 laser can be used to destroy this abnormal tissue band, increasing the caliber of the urethral meatus and improving symptoms. This review details the diagnostic workup of these disorders and describes the surgical management using the CO 2 laser.

Eight patients underwent carbon dioxide (CO 2) laser excision of benign cervical tumors during a 5-year period. Six patients had symptoms of abnormal vaginal bleeding, and 2 had asymptomatic cervical masses. All but one procedure was performed as outpatient surgery. Blood loss was minimal in 7 patients and less than 50 ml in the other patient. There were no cases of morbidity, and long-term follow-up showed excellent results. The technique to perform CO 2 laser excision of large benign cervical tumors is described.

Our previous studies demonstrated TRH stimulation of TSH beta gene transcription in rat pituitary cell cultures and in transient expression assays, with the TRH-sensitive region located between -1.3 kilobases and -204 basepairs (bp) relative to the major transcriptional start site. Using nuclear runoff and transient expression assays, we have analyzed the interactions among TRH, the phorbol ester 12-myristate 13-acetate (PMA), and the adenylate cyclase activator forskolin on TSH beta gene transcription. In cultured pituitary cells, TSH beta gene transcription was stimulated by 2 h of 10(-9) M TRH (2- to 4-fold), 100 nM PMA (2- to 6-fold), or 2 microM forskolin (1.5- to 2.5-fold) treatment, with additive interactions among all three effectors. Chimeric plasmids containing various 5'-flanking portions of the TSH beta gene and both transcriptional start sites, fused to the chloramphenicol acetyltransferase (CAT) gene, were transfected into the clonal pituitary GH3 cell line to delineate DNA sequences conferring this regulation. Transfected TSH beta CAT constructs containing TSH beta gene sequences from -2100/+27I150, -1295/+27I150, and -520/+27I150 expressed CAT enzyme activity which was stimulated by 24 h of TRH (2- to 3-fold), PMA (3- to 6-fold), or forskolin (1.5- to 3-fold) treatment, similar to observations in normal pituitary cells. In addition, a CAT expression vector construct containing only upstream TSH beta gene sequences from -703 to -85 bp, fused to the heterologous thymidine kinase promoter (tkCAT), exhibited similarly stimulated transcription in a transfection assay in response to TRH, PMA, and forskolin.(ABSTRACT TRUNCATED AT 250 WORDS)

In a prospective, randomized trial, 103 women underwent a total of 203 antenatal stress tests. One hundred four nipple stimulation contraction stress tests (BSTs) and 99 oxytocin challenge tests (OCTs) were performed. The patient populations were similar for the two groups. Uterine hyperstimulation with abnormal fetal heart rate patterns occurred with 2.9% and 1% of the BSTs and OCTs, respectively. The failure rate for the BST group was 22%. Maternal age and weight, parity and gestational age were not associated with test failure. Only one patient failed more than one BST, but she did not fail every such test. When test time (time from initiation to completion of the test) was compared between the two groups, a significant difference was found when the BST was successful. However, when test time in the total BST group (successful and unsuccessful BSTs followed by an OCT was compared to that in the OCT group, no difference could be found. Though an OCT following an unsuccessful BST took longer to perform than did a primary OCT, the difference was not statistically significant. It appears that BST test failure may relate more to the technique of nipple stimulation used than to intrinsic patient factors.

Bilateral tubal pregnancies are a rare variety of ectopic gestation. Tubal sterilization has become an important cause of tubal pregnancy. To our knowledge, this is the first case report of a twin gestation resulting in bilateral tubal pregnancies after a modified Pomeroy tubal sterilization.

Interstitial pregnancy is an uncommon variety of ectopic gestation. Rupture tends to occur later than with tubal gestations, and is often associated with catastrophic hemorrhage. Few case reports of term interstitial pregnancies have been reported, and none have been associated with fetal survival. This is the first reported case of a term interstitial pregnancy resulting in a live infant.

Few other specialists in medicine are as exposed to body fluids as obstetricians. Anxiety among these health care providers is therefore considerable. Guidelines for universal infection precautions are aimed at minimizing the risk of human immunodeficiency virus transmission by the creation of adequate physical barriers between the patient and the health care provider. Some of these guidelines are mutually exclusive to currently popular childbirth techniques in the very practical sense. However, the more important impact of universal infection precautions is likely to be psychological. Anxiety and the necessity for physical isolation are probably the worst basis for an empathic human relationship with the patient. It is the current challenge of obstetrics to maintain a sensitive and client-centered approach while judiciously dealing with the acquired immunodeficiency syndrome epidemic.