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Morphological effects of diethylstilbestrol on neonatal mouse uterus and vagina.

Administration of diethylstilbestrol (DES) to neonatal mice causes a high incidence of vaginal adenosis comparable to adenosis in women who were exposed to DES in utero. In the present study, scanning electron microscopy was used to investigate morphological events that might be involved in the development of adenosis. The effects of DES on luminal epithelia of neonatal mouse uterus, vaginal canal, and vaginal fornix were compared with the effects of the drug on those tissues of the adult mouse. BALB/c mice were given five s.c. injections of 2 micrograms DES in sesame oil or of sesame oil alone (controls) on postnatal Days 1 through 5 and were killed on Day 7. Ovariectomized adult BALB/c mice were given s.c. injections of 2 or 20 micrograms DES or of oil on 5 consecutive days and were killed 2 days after the last injections. In neonatal and adult uteri, DES stimulated the growth of microvilli on epithelial cells. The numbers of long and intermediate-length microvilli were markedly increased relative to those in control uteri. DES caused epithelial cornification in all samples of adult vaginal canal and fornix and in three of eleven Day 7 vaginal canal samples, but not in the remaining eight Day 7 vaginal canal samples or in any of ten Day 7 fornix samples. In the latter tissues, DES caused a marked increase in numbers of short, long, and intermediate-length microvilli on the apical surfaces of epithelial cells. Thus, in the majority of cases, en Day 7 vaginal canal samples, but not in the remaining eight Day 7 vaginal canal samples or in any of ten Day 7 fornix samples. In the latter tissues, DES caused a marked increase in numbers of short, long, and intermediate-length microvilli on the apical surfaces of epithelial cells. Thus, in the majority of cases, en Day 7 vaginal canal samples, but not in the remaining eight Day 7 vaginal canal samples or in any of ten Day 7 fornix samples. In the latter tissues, DES caused a marked increase in numbers of short, long, and intermediate-length microvilli on the apical surfaces of epithelial cells. Thus, in the majority of cases, the response of neonatal fornicovaginal epithelium to DES resembled that of neonatal and adult uterine epithelium more than it resembled the response of adult fornicovaginal epithelium; the uterine-like response might be causally related to the development of adenosis.

Age Factors↗

Measurement of diethylstilbestrol in plasma from patients with cancer of the prostate.

A specific radioimmunoassay has been developed for diethylstilbestrol (DES), using an antiserum raised against DES monocarboxymethyl ether and a tritium-labeled radioligand. Prior to radioimmunoassay, a fraction enriched in DES is obtained from a dichloroethane extract of plasma using Sephadex LH-20. The specificity of the assay is good, and the sensitivity (130 pg/ml) is adequate for accurate determination of DES in plasma from prostatic cancer patients treated with the drug. The precision is satisfactory, with an interassay coefficient of variation of approximately 10% at concentrations of approximately 1 ng/ml, and the blank values are negligible. Excellent agreement (r = 0.96) is observed between data obtained by radioimmunoassay and those obtained by a procedure using gas chromatography-high-resolution mass spectrometry. DES concentrations in the plasma of six treated (1 mg DES three times daily) patients were in the range 0.15 to 6.0 ng/ml. Increases in plasma concentration were observed within 2 hr of administration, with secondary rises occurring 5 to 6 hr later. Plasma testosterone concentrations were low in four of the patients; in a single subject, relatively high levels of testosterone were further elevated following administration of luteinizing hormone-releasing hormone.

Circadian Rhythm↗

Natural killer cell activity and tumor susceptibility in female mice treated neonatally with diethylstilbestrol.

Female NMRI or AKR/J mice were given daily s.c. injections of 5 micrograms diethylstilbestrol (DES) in 0.025 ml olive oil, or of olive oil only, for the first 5 days after birth. At the age of 6 to 7 weeks, both DES-treated females and control females were killed, and the cytotoxic activity of the spleen cells against standard natural killer cell target YAC-1 cells as well as the natural killer cell-sensitive I-522 cells and relatively insensitive I-51 AKR lymphomas were tested. The cytotoxic activity against I-51 cells was similar for DES-treated and control females while the DES-treated females had only about one-half the cytotoxicity activity to I-522 and YAC-1 cells as did controls. Control females eliminated radioactivity derived from 125I-labeled YAC-1 and I-522 target cells injected i.v. faster than did DES-treated females, while the results were similar for both animal groups when using I-51 cells. The cumulative death incidence was higher for DES-treated females than for control females after incubation with low numbers of I-522 cells but similar for both groups when using I-51 cells. Finally, the incidence of females developing methylcholanthrene-induced sarcomas, using a simple low-dose injection (10 or 20 micrograms), was higher among DES-injected animals than among controls. Taken together, the results indicate that female mice treated neonatally with DES have a functionally defective natural killer cell population, resulting in increased tumor susceptibility.

Animals↗

Cytologic problems in diagnosis of endocervical atypia in young females with and without maternal history of diethylstilbestrol exposure.

Sixteen cytologic endocervical atypias are reported with corroborative tissue analysis. Maternal diethylstilbestrol (DES) history was positive in three patients, one with clear cell adenocarcinoma, one with colposcopy and biopsy proven benign vaginal adenosis, and one with squamous carcinoma in situ of the cervix. Colposcopy was performed in two cases, one with adenosis and one with clear cell adenocarcinoma.In three patients, aged 19, 17, and 25, respectively, clear cell adenocarcinomas were found with unknown DES history and positive cytology. In five other cytologically positive cases, endocervicitis was found in three, atypical endocervical hyperplasia in one, and severe squamous dysplasia in one. This represents a cytologic false positive or specificity reading error of 37 percent in this type of material. One case of clear cell adenocarcinoma was linked with DES exposure.In the six suspicious and borderline cytologic reports, one vaginal adenosis, one condyloma acuminata, and one squamous carcinoma in situ were found. This represents a false positive or specificity reading error rate of 43 percent.Cytology is clinically valuable in Proven cases of vaginal adenosis or adenocarcinoma. It is not a diagnostic modality, with a high sensitivity error (false-negative rates) of 50 percent and higher, as reported in the literature and high specificity error (false-positive rates) as herein reported.

Adolescent↗

Localization of 3H-diethylstilbestrol in skeletal muscle.

The localization of diethylstilbestrol (DES) in skeletal muscle was studied in CF1 mice and perfused rat hindlimbs. There was a slow accumulation of 3H-DES in mouse muscle from 4 to 24 hours following i.p. injection even though plasma DES was decreasing. Twenty-four hours after injection of 50 microCi 3H-DES (714 pmole) mouse gastrocnemius contained 8.9 x 10(-17) mole unaltered 3H-DES per mg muscle. Extrapolating to the entire skeletal muscle mass of the animal, this represents 0.15% of the radioactivity injected. The radioactivity in muscle was completely extracted with 95% ethanol or ether: ethanol (3:1), and both unaltered DES and DES-metabolites were present in the extracts. The fraction of radioactivity due to unaltered DES 4 hours after injection was 0.51 +/- 0.09 in muscle and 0.30 +/- 0.11 in plasma. Significant extrahepatic metabolism of DES was demonstrated in perfused isolated rat hindlimbs by the presence of DES-metabolites in the perfusate. The radioactivity extracted from the perfused muscle itself was unaltered DES. These results indicate that skeletal muscle is an important site of DES localization in rodents.

Animals↗

Diethylstilbestrol di-trimethylacetate (DSTMA). A long-acting estrogen.

A series of estradiol and diethylstilbestrol (DES) esters was prepared as part of a study of the structure-activity relations in estrogens. Among the esters tested, DES di-trimethylacetate (DSTMA) exhibited the most favorable combination of a low minimum-effective dose and prolongation of effect in all experimental groups (mice, rats, dogs). Toxicity of DSTMA was low. A clinical trial in four stage III and stage IV prostatic cancer patients showed that dosage levels of DSTMA as high as 75 mg per day for short periods were well tolerated and no toxic symptoms attributable to the estrogen itself were observed.

Acetates↗

Observations on the psychological impact of diethylstilbestrol exposure and suggestions on management.

The emotional impact of diethylstilbestrol (DES) exposure is described in a series of 50 mothers and daughters interviewed by psychiatrists. Patterns of response to this trauma and methods of resolution are discussed, and opportunities for preventive intervention by gynecologists are suggested. Specific, open dialogue about DES with the patient as a colleage can minimize the emotional sequelae of the experience.

Affective Symptoms↗

Myelodyspoietic syndrome associated with diethylstilbestrol therapy.

We describe a patient with a myelodysplastic syndrome characterized by pancytopenia and an excess of myeloblasts in the bone marrow. He had received massive doses of diethylstilbestrol (150 mg daily) for seven years as therapy for prostatic carcinoma. Although myelodyspoiesis has been associated with other drugs, a relationship to estrogen therapy in man has not been reported previously. However, the administration of estrogens to animals has produced pancytopenia with a relative monocytosis and vacuolization of leukocytes. Examination of bone marroa, erythroid hypoplasia, and, with prolonged therapy, aplastic anemia. Further animal studies have demonstrated that estrogens exert a suppressive effect on marrow stem cells and granuloid progenitor cells. These experimental observations suggest a possible role of estrogens in the genesis of the hematologic changes we observed.

Aged↗

Urinary tract changes associated with exposure in utero to diethylstilbestrol.

Intravenous pyelograms were performed on 128 women who had been exposed in utero to diethylstilbestrol (DES). For comparison, intravenous pyelograms (IVPs) of 48 non-exposed women were reviewed: No difference in urinary tract abnormalities was noted. There was no significant increase in urinary tract abnormalities in women with upper genital tract abnormalities demonstrated by hysterosalpingogram as compared to a group of women whose x-rays of the uterus were normal.

Abnormalities, Drug-Induced↗

Immune responsiveness of adult mice exposed neonatally to diethylstilbestrol, steroid hormones, or vitamin A.

Neonatal hormone treatments with estradiol, progesterone, testosterone, cortisol, or especially diethylstilbestrol (DES) were found to alter the immune responsiveness of spleen cells from female mice. No significant change in immunocompetence occurred in neonatally vitamin A-treated animals. In general, the hormone treatments caused an initial hyperstimulation of spleen cell response in 1-month-old mice, followed by a gradual decline in immune responsiveness from about 2 to 9 months of age. This hyperstimulation followed by a gradual loss of response was most apparent in the DES-treated group, and to a less extent in the testosterone and cortisol groups. Spleen cells from mice treated neonatally with estradiol or progesterone produced decreased graft-versus-host responses in the lymph node assay but were normal in all other immunological tests.

Animals↗

The ultrastructure of some histopathological changes seen in oviducts of mice continuously fed diets containing diethylstilbestrol.

Selected oviductal lesions associated with the continuous feeding of diethylstilbestrol (DES) were examined ultrastructurally Virgin female mice were fed diets containing 0, 320 and 640 ppb DES from 4 weeks of age. All animals reported on in this study were removed from the study and sacrificed for histopathological and electron microscopy examination when moribund and ranged from 622 to 762 days on the experiment. One oviductal change consisted of markedly vacuolated ciliated epithelial cells located primarily in the fimbria. Electron microscopy revealed that the large vacuoles were present throughout the cytoplasm of affected cells. These vacuoles were not lipid and appeared to be fluid-filled. No significant alterations in number or structure of cilia were observed. Since this alteration was seen in both DES and control animals, it may be age-related. A second kind of oviductal lesion consisted by enlarged secretory cells located predominantly in the isthmus. The cytoplasm of these cells consisted of extremely enlarged, dilated rough endoplasmic cisternae forming a subnuclear mass. Numerous primary and secondary lysosomes were seen in the apical cytoplasm between the cell membrane and nucleus. No evidence of a Golgi complex was seen. These alterations were seen only in DES-exposed animals and seem to indicate continued protein synthesis by the secretory cells but a deficiency in the packaging of this protein into secretory granules.

Animals↗

Evolution of diethylstilbestrol-associated genital tract lesions.

The evolution of the colposcopic manifestations of adenosis involving the cervicovaginal hood and vagina was determined by the long-term follow-up of 173 diethylstilbestrol-exposed women. Serial gross observations in conjunction with histopathologic data showed that adenosis is a dynamic lesion that undergoes progressive squamous metaplasia over time. Ultimately, adenosis may disappear completely, to be replaced by normally glycogenated mucosa. A schema for the evolution of adenosis is presented.

Cervix Uteri↗

Diethylstilbestrol-induced upper genital tract abnormalities.

In utero diethylstilbestrol (DES) exposure has recently been associated with apparently unique abnormalities of the upper genital tract. Utilizing a standardized technique of hysterosalpingography (HSG) and a linear planimeter, the following measurements were made in a group of 13 DES-exposed women and compared with a control group of 22 women undergoing HSG during infertility investigations (mean +/- standard error: endometrial cavity area, 323.23 +/- 32.13 sq mm versus 626.56 +/- 52.75 sq mm; endometrial cavity circumference, 128.65 +/- 5.08 mm versus 140.52 +/- 5.56 mm; upper uterine segment length, 28.80 +/- 1.11 mm versus 38.03 +/- 1.81 mm; lower uterine sugment length, 36.03 +/- 4.77 mm versus 42.24 +/- 2.39 mm; intercornual distance, 36.40 +/- 2.56 mm versus 38.25 +/- 1.58 mm; internal os diameter, 3.90 +/- 0.44 mm versus 4.43 +/- 0.23 mm; widest diameter of the endocervical canal, 3.78 +/- 0.40 mm versus 9.39 +/- 0.60 mm; isthmic tubal diameter, 1.25 +/- 0.08 mm versus 1.20 +/- 0.01 mm; ampullary tubal diameter, 4.79 +/- 0.43 mm versus 4.65 +/- 0.22 mm. The endometrial cavity area, upper uterine segment, and endocervical canal measurements were significantly smaller in the DES-exposed group (P less than 0.01). The upper genital tract abnormalities observed appeared to be unlike spontaneously occurring Müllerian malformations and correlated with DES-induced cervicovaginal changes. Primary dysmenorrhea and menstrual irregularity occurred in 40% and 47%, respectively, of the DES-exposed patients.

Abnormalities, Drug-Induced↗

Structural and functional abnormalities in the sex organs of male offspring of mothers treated with diethylstilbestrol (DES).

The in utero effects of DES (Diethylstilbestrol) on the human male genital tract are reported in this follow-up study of male offspring of DES-treated mothers. Both anatomical and functional abnormalities were significantly greater in the DES-exposed males as compared to control males whose mothers were all participants in a prospective, randomized double blind study of the effects of DES on pregnancy at the Chicago Lying-in Hospital during the early 1950s. Epididymal cysts, hypotrophic testes and capsular induration of the testes were among the more common genital lesions found in 27% of 134 DES-exposed males as compared to a 7% incidence in 119 control males. Spermatozoa analyses revealed severely pathologic changes (Eliasson score greater than 10) in 29% of 28 DES-exposed males and 0% of 18 control males (with or without genital i.e., physical abnormalities). Abnormal findings on physical examination combined with sperm abnormalities (Eliasson score greater than or equal to 5) were found in 29% of DES-exposed males versus 0% of 18 control males. Cytologic examinations did not reveal malignant cells from the following materials: urines before and after prostatic massage or ejaculation, prostatic fluids and aspirates from epididymal cysts.

Abnormalities, Drug-Induced↗

[Study on prevention of flare-up phenomenon following initial LH-RH analogue administration: combination therapy with diethylstilbestrol].

To eliminate the initial testosterone (T) surge and prevent the risk of flare-up induced by the first administration of luteinizing hormone-releasing hormone (LH-RH) analogue, we examined the effectiveness of short-term combination therapy with diethylstilbestrol (DES). Sixteen previously untreated patients with prostate carcinoma (Stage C: 4 cases, Stage D2: 12 cases) were randomly assigned to 4 groups. Each group included 4 patients. Group 1 received DES (500 mg daily injection) for 7 days, staring before the first subcutaneous injection of luteinizing hormone releasing hormone (LH-RH) analogue (3.6 mg of Zoladex depot); Group 2 received 500 mg of DES injection for 7 days prior to the initiation of therapy with the LH-RH analogue, and moreover received a DES injection for 7 days; Group 3 received 500 mg of DES injection for 7 days simultaneously with the first injection of the LH-RH analogue; Group 4 was pretreated with 500 mg of DES injection for 7 days prior to the initiation of therapy with the LH-RH analogue, with additional DES injection for 3 days. In groups 1, 2 and 4 the level of T decreased during the 7 days of DES therapy, and at 3 days after the first injection of LH-RH analogue increased again. The mean T value in groups 1, 2 and 4 decreased by 33.2 +/- 27.6% (mean +/- SE), 20.5 +/- 20.8%, 13.2 +/- 9.8%, respectively, at the day of the first injection of LH-RH analogue, and increased by 75.2 +/- 21.6%, 70.7 +/- 63.4%, 56.7 +/- 46.4%, respectively, at 3 days after the first injection of LH-RH analogue.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenocarcinoma↗

Endometrial integrin expression in women exposed to diethylstilbestrol in utero.

OBJECTIVE: To determine if the endometrium of women exposed in utero to diethylstilbestrol (DES) demonstrates altered integrin expression compared with normal fertile controls. DESIGN: Case control study. Expression of eight integrins in 15 luteal phase endometrial biopsies from DES-exposed women were compared with 17 biopsies from age- and cycle day-matched controls. All patients were ovulatory. Endometrial biopsies were performed in the luteal phase, and all were histologically "in phase." SETTING: Infertility practice of an academic teaching hospital. PATIENTS: Women exposed in utero to DES and matched fertile controls. MAIN OUTCOME MEASURES: Intensity of endometrial integrin immunohistochemical staining by the semiquantitative HSCORE technique. RESULTS: Endometrial stroma of DES-exposed women demonstrated greater expression of the integrin subunits alpha 5 and alpha v. No differences were noted between DES-exposed women and fertile controls in the glandular epithelial expression of integrin subunits alpha 1, alpha 2, alpha 3, alpha 6, and alpha v nor the stromal staining of subunits of alpha 6 and beta 3. Epithelial expression of integrin subunits beta 3 and alpha 4, reliable luteal markers of uterine receptivity, were similar in DES and control patients. CONCLUSIONS: Markers of uterine receptivity are similar in the endometrium of women exposed in utero to DES compared with normal fertile controls. Interesting differences exist in stromal integrin expression between groups, though the significance of this finding is uncertain at present. It is unlikely that DES-exposed women have significantly altered endometrial function as a cause of infertility.

Adult↗

Diethylstilbestrol metabolism in food-producing animals.

The metabolic fate of diethylstilbestrol (DES) in food-producing animals is described in this paper. Emphasis is on ruminant animals because DES is currently used in their production. Data presented from recent studies in ruminants show 14C excretion (rates and routes) and distribution after ingestion or subcutaneous implantation of 14C-labeled DES. The chemical nature of the radioactive compounds in excreta and liver is described. In addition, DES metabolism in cockerels and swine and degradation of DES in feces are discussed. The sources of data in this review are published studies, and a limited amount of unpublished data from the author's laboratory are included.

Animals↗

Exposure to diethylstilbestrol during a critical developmental period of the mouse reproductive tract leads to persistent induction of two estrogen-regulated genes.

Exposure to estrogens during critical periods of development induces teratogenic and carcinogenic lesions in the reproductive tracts of humans and experimental animals. It is important to determine the molecular and cellular targets of estrogenic chemicals and to establish the mechanisms by which interactions of estrogens with the developing genital tract results in permanent lesions of growth and differentiation. The experiments presented here were designed to examined the effects of neonatal estrogen exposure on the expression of two genes, lactoferrin and epidermal growth factor, that are subject to steroid hormone regulation. Using in situ and Northern RNA hybridization, immunoblotting, and immunohistochemistry, our data demonstrate that exposure to the synthetic estrogen, diethylstilbestrol, during a critical neonatal period results in the persistent ovary-independent induction of mRNA and protein encoded by these two genes in the mouse uterus and vagina. The constitutive expression of lactoferrin and EGF, and probably other estrogen-regulated genes, may contribute to the establishment of a permanently "estrogenized" phenotype which is then instrumental in the development of abnormal tissue morphogenesis, function, and neoplasia in the rodent reproductive tract.

Animals↗