Pseudorosette-Like Proliferations of the Endometrium.

The endometrium displays a wide spectrum of appearances in both neoplastic and non-neoplastic tissue. Unusual proliferations are not infrequently encountered and may lead to misinterpretation. In this study, we investigated pseudorosette-like proliferations (PLPs) found within the endometrial stroma which, to our knowledge, have not been previously reported. Nineteen endometrial samples with PLPs were identified over a period of 5 yr. Characteristics of the endometrium in which the PLPs were arising as well as patient information were recorded for each case. In addition, residual tissue from 10 of the cases was immunostained for cytokeratin, CD10, smooth muscle actin, S-100, and caldesmon to better characterize the lesions. In all cases the endometrium was in the proliferative phase and none of the patients reported the use of exogenous hormones. In 89% (17 of 19) of the cases, PLPs were present as a single focus; 2 cases showed multiple PLPs. Of the 10 immunostained cases, 90% (9 of 10) showed strong/diffuse staining for smooth muscle actin. Six cases showed negative or weak (1+) staining for CD10, whereas 3 showed moderate (2+) and 1 showed strong (3+) staining. In all cases the PLPs were negative for cytokeratin AE1/AE3, S-100, and caldesmon. These studies suggest that PLPs are benign proliferations with smooth muscle differentiation.

Action of erythropoietin in vitro on rabbit reticulocyte membrane Ca2+-ATPase activity.

The mechanism of action of erythropoietin is thought to require specific interaction with the target cell surface and involve alteration of cellular calcium metabolism. Using the rabbit reticulocyte membrane as a model of the immature red cell membrane, we investigated the effects of human recombinant erythropoietin on membrane Ca2+-ATPase (calcium pump) activity in vitro. Erythropoietin in a concentration range of 0.025 to 3.0 U/ml progressively decreased membrane Ca2+-ATPase activity by up to 64% (P less than 0.01). These concentrations have been shown by others to stimulate in vitro erythroid growth. The action of erythropoietin on reticulocyte Ca2+-ATPase required an incubation time of 1 h before enzyme assay for maximum effect and was neutralized by antierythropoietin antiserum. Other nonhemopoietic growth factors (epidermal growth factor, insulin) had no effect in this assay. Ca2+-ATPase activity of membranes prepared from rabbit mature red blood cells was not inhibited by erythropoietin. The novel effect of erythropoietin on reticulocyte membrane Ca2+-ATPase activity is a mechanism by which erythropoietin can influence cellular Ca2+ metabolism.

Ultrastructural and morphometric study of diethylstilbestrol-associated lesions diagnosed as cervical intraepithelial neoplasia III.

Investigations were carried out to characterize diethylstilbestrol (DES)-associated squamous lesions and to assess their biological significance. Five DES-associated cervical lesions displayed architectural features which were diagnosed as cervical intraepithelial neoplasia (CIN) III, such as full-thickness replacement by atypical squamous cells with vertical orientation and absence of normal polarity. Electron microscopic examination revealed only one of the five to be consistent with the generally recognized ultrastructural picture of non-DES CIN III. In the remaining four lesions, the moderate-to-large numbers of tonofibrils and well-developed desmosomes distinguished them from the true CIN III lesions. Morphometric studies indicate the five DES-associated lesions in this study as a group to be significantly different from normal squamous epithelium, from maturing metaplasia, and from non-DES-related CIN III in the parameters of differentiation studied. Their intermediary position between maturing metaplasia and non-DES CIN III suggests that they are more differentiated than CIN III and less differentiated than maturing metaplasia. Nuclear area measurements indicate the increased nuclear-cytoplasmic ratio observed in the DES-associated CIN III lesions of this study is due to a decrease in cytoplasmic volume, as opposed to an increased nuclear size.

PIP: 5 diethylstilbestrol (DES)-associated squamous lesions, all of which had been diagnosed as cervical intraepithelial neoplasia 3 (CIN 3), were investigate to assess the biological significance of these lesions among women exposed in utero to DES. The diagnoses were based on characteristics such as full-thickness replacement by atypical squamous cells with vertical orientation and absence of normal polarity. However, by electron microscopic techniqeus it was found that only 1 of the 5 lesions was consistent with the generally recognized ultrastructural look of non-DES-associated CIN 3. The remaining 4 leasions had in architecture moderate-to-large numbers of tonofibrils and well-developed desmosomes which distinguished these lesions from true CIN 3 lesions. A morphometric study indicated that the 5 DES-associated lesions studied here are significantly different from normal squamous epithelium, from maturing metaplasia, and from non-DES-related CIN 3, suggesting that they are more differentiated than CIN 3 and less differentiated than maturing metaplasia. When nuclear areas were measured in these lesions, increased nuclear-cytoplasmic ratio was observed in the DES-associated CIN 3 lesions, caused by decreased cytoplasmic volume rather than increased nuclear size.

Combined effects of 1,25-dihydroxyvitamin D3 and platinum drugs on the growth of MCF-7 cells.

The effects of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and platinum treatments (both singly and combined) on the growth inhibition of MCF-7 cells, an epithelial cell line shown to possess specific receptors for 1,25(OH)2D3, were evaluated. The inhibitory effects of 1,25(OH)2D3 and platinum on MCF-7 cell proliferation in vitro were time and dose related. The data showed that 10 nM and 100 nM 1,25(OH)2D3 inhibited MCF-7 cell growth by 10.8 +/- 2.4% and 34.9 +/- 0.5% (mean +/- SE), respectively. The degrees of growth inhibition induced by 0.2 to 200 micrograms/ml of cis-diammine-1,1-cyclobutane dicarboxylatoplatinum(II) (carboplatin) were slightly less than those induced by 0.02 to 20 micrograms/ml of cis-diamminedichloroplatinum(II) (cisplatin). The combined administration of 10 nM and 100 nM 1,25(OH)2D3 with either carboplatin (200 to 0.2 micrograms/ml) or cisplatin (20 to 0.02 micrograms/ml) was evaluated. Addition of 1,25(OH)2D3 to the platinum resulted in marginal to marked enhancement of growth inhibition over that observed with either platinum alone. The strength of these interactions varied inversely with the dose of the platinum drugs. Evaluation of drug interactions with isobolograms showed that at near-serum levels, carboplatin or cisplatin interacted synergistically with 1,25(OH)2D3 to inhibit MCF-7 cell growth. Our findings suggest potential usefulness in combining 1,25(OH)2D3, a biological modifier, with cytotoxic agents for the treatment of malignant disease.

KRAS Mutations in Mucinous Lesions of the Uterus.

OBJECTIVES: The current study examined the KRAS mutation status in a spectrum of mucinous lesions of the uterus, including mucinous metaplasia (MM), atypical mucinous proliferation (AMP), endocervical mucosa, and microglandular hyperplasia (MGH). METHODS: Thirty-nine cases, including 15 AMPs, nine MMs, nine MGHs, and six normal endocervical mucosas, were selected from the departmental archive. All AMP cases with follow-up biopsies or hysterectomies were reviewed. Genomic DNA was extracted from formalin-fixed, paraffin-embedded tissue and KRAS codons 12 and 13 sequence analyzed. RESULTS: KRAS codon 12 and 13 mutations were detected in 10 (67%) of the 15 AMP cases. No KRAS mutations were identified in MMs, MGHs, and endocervical mucosas (P = .002, AMP vs MM or MGH, Fisher exact test). Most women with AMP were postmenopausal (13/15 [86.7%]) and presented with dysfunctional uterine bleeding. Among the 10 cases of AMP harboring KRAS mutations, six (60%) cases were subsequently diagnosed with carcinoma, one with atypical complex hyperplasia, and two with AMP within endometrial polyps. CONCLUSIONS: The results suggest a possible association between KRAS mutations and mucinous differentiation in endometrial carcinogenesis. KRAS status can help in assessing benign from precursor or malignant mucinous lesions as well as differentiate endometrial lesions from those of cervical origin.

Structure-activity relationships of sex steroid analogs determined in vitro in a thyroid hormone-responsive membrane Ca2+-ATPase model.

Physiological concentrations of L-T4 were found previously to stimulate Ca2+-ATPase activity in vitro in reticulocyte membranes from female rabbits and to inhibit this enzyme in reticulocyte membranes from males. In these previous studies, preincubation of intact cells or ghosts with testosterone (5 X 10(-11) M) converted female-source reticulocyte membranes to male-type responsiveness to thyroid hormone (inhibition of Ca2+-ATPase activity). Preincubation of reticulocyte membranes with 17 beta-estradiol (5 X 10(-11) M) converted male-source membranes to female-type responsiveness (stimulation by L-T4 of membrane Ca2+-ATPase activity). Using this sex steroid-sensitive thyroid hormone-dependent membrane enzyme system, we investigated the structure-activity relationships of analogs of sex steroids and unrelated steroids. 5 beta-Androstanes were active compared to testosterone in assays using female-source membranes, while 5 alpha-androstanes were less active. Within the 5 beta-androstanes, activity was dependent on at least one hydroxyl group at the C3- or C17-position. Nongonadal steroids tested were less active, establishing specificity of the sex steroid effect in assays using female-source membranes. Assayed in male-source membranes, estrone and 3-hydroxy-1,3,5-(10)7-estratraen-17-one (equilin) were active compared for estrogen effect with 17 beta-estradiol, while estriol was less active. The activities of hydrocortisone and aldosterone were 76% and 71%, respectively, in this system. These structure-activity relationships are distinct from those described for gonadal steroid-cytoplasmic binding proteins or nuclear interactions, and represent a novel sex steroid-thyroid hormone effect on activity of a membrane enzyme.

Juvenile granulosa cell tumor of the infantile testis. A report of 14 cases.

Fourteen testicular tumors diagnosed in infants less than 6 months of age had a distinctive appearance resembling that of the juvenile granulosa cell tumor of the ovary. One of them was discovered at autopsy in an infant of 30 weeks' gestational age; seven were diagnosed during the first few days of life, and the remainder between 3 weeks and 4 1/2 months of age. Enlargement of a scrotal testis was the presenting manifestation in 10 cases and abdominal swelling in one case; one tumor was found in a descended testis that had undergone torsion and one was discovered in an inguinal testis at the time of inguinal herniorrhaphy. The tumors ranged from 0.8 to 5.0 cm in diameter and were cystic or partly cystic and partly solid. Microscopic examination disclosed both follicular and solid components. The follicles were of varying sizes and contained eosinophilic or basophilic fluid that stained positively for mucin in some of the cases. The solid foci typically had a nodular arrangement and occasionally were hyalinized or had a basophilic background due to the presence of intercellular mucin. The neoplastic cells contained moderate to large amounts of eosinophilic cytoplasm and round to oval hyperchromatic nuclei, which typically lacked grooves. The mitotic rate varied from less than 1-24/10 high-power fields. Limited follow-up examination revealed no evidence of recurrence. The microscopic features of these neoplasms warrant their designation as "juvenile granulosa cell tumor."

Juvenile granulosa cell tumor--another neoplasm associated with abnormal chromosomes and ambiguous genitalia. A report of three cases.

Three infants, 3 months of age or younger with abnormal karyotypes and ambiguous genitalia, had gonadal juvenile granulosa cell tumors. Two of the patients had mixed gonadal dysgenesis and the third had an intersexual disorder of undetermined type. Two tumors arose in undescended testes, and the third in an undescended gonad of uncertain nature. The occurrence of this uncommon neoplasm in these infants indicates that it is another type of neoplasm that may develop in the gonad of a patient with an abnormal karyotype and ambiguous genitalia.

"Placental polyp": light microscopic and immunohistochemical observations.

A case of the chronic type of placental polyp, occurring in a 37-year-old woman approximately 9 years after abortion of her last known pregnancy, is reported. The placental polyp was predominantly composed of necrotic and hyalinized chorionic villi without identifiable lining trophoblast; however, some villi showed a thin rim of apparently viable syncytiotrophoblast that exhibited focal strong positivity for human chorionic gonadotropin by immunohistochemical studies. Intermediate trophoblast, especially abundant within the intervillous fibrin, appeared most viable and showed strong positivity for human placental lactogen (hPL); syncytiotrophoblast also showed focal positivity for hPL. The basal aspect of the polyp was composed of abundant decidua that contained dilated and ectatic blood vessels. This study demonstrates the presence of cytoplasmic markers for pregnancy in a chronic type of placental polyp, apparently of 9 years' duration, and draws attention to an entity that may be encountered more frequently due to the current prevalence of induced abortions.

Extratubal secondary trophoblastic implants: clinicopathologic correlation and review of the literature.

Extratubal secondary trophoblastic implants (ESTI) are a rare complication of conservative laparoscopic procedures for tubal ectopic pregnancies. These implants present with persistent beta-hCG titers postoperatively and are probably the result of disruption of the ectopic pregnancy at salpingostomy or morcellation of the fallopian tube at salpingectomy. We describe the case of a 32-year-old woman who underwent a laparoscopic salpingectomy for a tubal ectopic pregnancy that was complicated postoperatively by peritoneal ESTI including extensive omental implants. Intraoperatively the lesions appeared as 0.3 cm red-black nodules and, microscopically, consisted of degenerating chorionic villi associated with implantation changes in the surrounding tissue. To the pathologist unaware of the clinical entity of ESTI, these lesions may present a diagnostic challenge.

Absence of teratogenic effects of progesterone on the developing genital tract of the human female fetus.

It has been questioned whether prenatal exposure to progesterone, like exposure to diethylstilbestrol (DES), results in teratogenic abnormalities of the upper and lower genital tract in human females. Through the use of a recently described model in which human fetal reproductive tracts can be transplanted and grown in vivo for extended periods in athymic nude mice, genital tracts from human female fetuses, ages 7 to 18 weeks postovulation, were grafted into castrated murine hosts and grown for 4 to 10 weeks in the presence or absence of continuous exposure to progesterone. Substantial growth was observed. For all specimens, the morphogenetic process proceeded normally, resulting in the harmonious organization of a complete, well differentiated genital tract composed of fallopian tubes, uterine corpus, and cervix and vagina. The fallopian tubes were highly convoluted and disclosed fimbria. The uterine corpus was lined by a simple columnar epithelium; two layers of stroma in the wall were distinctly separated from each other. In the older specimens, the outer layer of stroma had assumed microscopic properties diagnostic of smooth muscle (myometrium). In the majority of specimens the region of the cervix/vagina disclosed the development of a fornix-like evagination at which point or slightly cranially there was a gradual but defined transition from columnar epithelium to squamous epithelium. The inner layer of endometrial stroma tapered and disappeared at or close to the squamocolumnar junction. The vaginal stroma was a single homogeneous layer and was continuous with the myometrium of the uterine corpus. In the context of this model system, prenatal exposure of the developing human female genital tract of progesterone was not associated with any obvious teratogenic effects.

Detoxifying enzymes in human ovarian tissues: comparison of normal and tumor tissues and effects of chemotherapy.

Many anticancer drugs exert their cytotoxic effects via formation of oxygen free radicals. Cellular thiols, glutathione (GSH)-dependent enzymes and other redox enzymes are involved in the metabolism of these anticancer drugs and of the oxygen free radicals that may be generated during their metabolism. We quantified these biochemical parameters in cytosol from human ovarian tissues. We compared non-protein thiol levels, GSH transferase, GSH peroxidase, superoxide dismutase, catalase, DT diaphorase and aldehyde dehydrogenase activity in serous ovarian tumors (n = 15), other malignant ovarian tumors (n = 12), benign ovarian tissue (n = 10) and histologically normal ovarian tissue (n = 12). Mean GSH transferase and DT diaphorase activities were similar in serous and other malignant ovarian tumors. GSH transferase activity was decreased in malignant tissues relative to normal and benign tissues. Mean DT diaphorase and superoxide dismutase activities were increased in the malignant tissues, although this was not statistically significant. The mean levels of all enzymes except superoxide dismutase and aldehyde dehydrogenase in benign tissues were fairly similar to the mean levels found in normal tissue samples. Tissues from patients with serous ovarian tumors, who had received cyclophosphamide and cisplatin prior to surgery, also were analyzed (n = 7). Except for aldehyde dehydrogenase, all the parameters measured were decreased in these samples relative to serous tissue from untreated patients. These biochemical analyses may be useful in understanding the mechanisms involved in the response to chemotherapy.

The immunohistochemical profile of malignant mixed müllerian tumor. Overlap with endometrial adenocarcinoma.

The distinction between malignant mixed müllerian tumor (MMMT) of the female genital tract and poorly differentiated endometrial adenocarcinoma (EA) is sometimes difficult and arbitrary. Although several studies have described the immunohistochemical profile of MMMT and EA, the results have varied, and controversy regarding the histogenetic relationship between them remains. The authors examined the histologic characteristics and immunohistochemistry of 31 formalin-fixed paraffin-embedded MMMTs from a variety of female genital tract sites with a panel of monoclonal and polyclonal antibodies using the avidin-biotin-peroxidase complex (ABC) method. Eighteen neoplasms were homologous and 13 were heterologous; the average patient age was 67 years. In all cases the epithelial component stained for keratin (K), whereas the stromal component stained in 48%; vimentin (V) was detected in the epithelial component in 35% and the stromal component in 81%. Myoglobin (M) detected rhabdomyoblasts in three of five cases tested; desmin (D) was found in five of six and actin (A) in all six cases with skeletal muscle. Overall, A stained the stromal components in 45% of cases, whereas none of the epithelial components stained. The coexpression of K and V in the epithelial component of 35% of cases and in the stromal component of 48% of cases led the authors to conclude that the immunoprofile of MMMT broadly overlaps with that of EA and, independent of morphologic findings, it is often not reliable or useful in distinguishing between the two. Furthermore, the variable immunoprofile suggests that MMMT may be a histogenetically heterogeneous group of neoplasms, including both carcinomas (metaplastic, sarcomatoid) as well as true carcinosarcomas. Although A detected rhabdomyoblasts in all cases, it also stained neoplastic mesenchymal elements in eight more cases, suggesting the presence of smooth muscle or myofibroblasts in the stroma of some MMMTs.

ADASP recommendations for processing and reporting of lymph node specimens submitted for evaluation of metastatic disease.

It is well known that different pathologists in different laboratories follow different protocols for the processing and examination of these specimens. There is also extensive literature (some of which is summarized in the references appended to the present report) on the likelihood of identifying metastases of varying sizes with different methods of preparation, as well as on the clinical significance of this identification, which varies not only from site to site but also from report to report on the same site. The Association of Directors of Anatomic and Surgical Pathology (ADASP) has reviewed this literature as well as the personal experience of its own members to present a set of recommendations for lymph node biopsies, lymph node dissections, sentinel node biopsies, lymph node fine needle aspiration (FNA) and core needle biopsies. It should be noted that these recommendations are intended specifically for lymph nodes being studied for metastatic neoplasms, and are not intended to apply to lymph nodes being evaluated for lymphoma, infections, and other disease processes. They are, however, formulated generically enough to apply regardless of whether the primary tumor is a carcinoma of the breast, carcinoma of the prostate, melanoma, or any other malignant, potentially metastasizing tumor. The Association has published numerous documents with recommendations for reporting surgical pathology specimens involving particular organ sites (for example, breast, pancreas, thyroid, etc.) However, the Association has not yet considered the generic question of dealing with lymph node specimens in which the intent is to search for and document the presence of metastatic disease. We are also unaware of guidelines for pathologists published by any other organization on this subject.

Endodermal sinus tumor of the vulva in an infant.

An endodermal sinus tumor arising in the vulva of a 22-month-old infant is reported and the features of 3 previously recorded endodermal sinus tumors of the vulva are reviewed. Despite radical surgery, radiation therapy, and chemotherapy the tumor was fatal within 6 months. Only one of the 3 previously described patients with this type of vulvar tumor has survived more than 2 years. That tumor was the smallest reported, being the only one under 2 cm in diameter.

Adenocarcinoma of the vagina in a patient with gonadal dysgenesis.

Adenocarcinoma of the vagina is an uncommon tumor. Many theories have been postulated for its etiology and tissue origin. Its occurrence in young women exposed in utero to diethylstilbestrol has pointed to estrogen influence as a possible etiologic factor. A case of adenocarcinoma of the vagina in a patient with gonadal dysgenesis is presented, and histologic and electron-microscopic observations are discussed. This patient is unique in that development of the vaginal neoplasm occurred in the absence of estrogen influence.

Increased dermal elastic fibers in the tight skin mouse.

OBJECTIVE: The tight skin (Tsk-1) mouse has been proposed as a model for systemic sclerosis on the basis of increased accumulation of collagen and glycosaminoglycans in the skin, and by the presence of serum autoantibodies. The genetic basis of the mutation has been identified as a genomic duplication within the fibrillin-1 (Fbn-1) gene that results in a larger than normal Fbn-1 transcript, but the mechanism that leads to dermal fibrosis is unclear Fibrillin molecules associate into a polymer that is coated with elastin molecules to form elastic fibers. To further evaluate the Tsk-1 mouse model of scleroderma, we have studied elastic fibers in the skin of these mice. METHODS: Skin sections obtained from C57BL/6-TSK+ (Tsk-1) and C57BL6-pa/+ (control) mice were stained with Masson's trichrome for evaluation of collagen and Gomori's aldehyde fuchsin stain for elastic tissue. Computer assisted image analysis was performed to quantify differences in histologic sections. RESULTS: Tsk-1 mice had a highly significant increase in the percentage of elastic fibers (19.6%) in the dermis compared to control mice (7.9%) [p < 0.001]. This correlates with the findings in the skin of systemic sclerosis patients where increased elastic fibers have been observed. In addition, an increased level of dermal collagen staining was also observed in the Tsk-1 dermis (82.9%) compared with the level in normal sections (73.7%) [p < 0.01]. CONCLUSION: These data support the use of the Tsk-1 mouse as a model for the connective tissue abnormalities of human scleroderma.

A matched study of surgically treated stage IB adenosquamous carcinoma and adenocarcinoma of the uterine cervix.

Thirty-eight patients with surgically treated stage IB adenosquamous carcinoma of the uterine cervix (AS) have been matched with patients with other histologic subtypes of adenocarcinoma (A) for stage, lesion size, node status, grade of adenocarcinoma and age at diagnosis. An additional six patients with AS were unable to be matched. Overall 5-year survival and disease-free survival for the matched AS and A were not significantly different, 83 vs. 90%, and 78 vs. 81% nor were the number of recurrences, 8/38 AS vs. 6/38 A, but the mean time to recurrence was significantly shorter in the AS group: 11 vs. 32 months (P = 0.003). A subgroup of AS with a high risk of a poor outcome can be identified based on either lesion size >/= 4 cm, depth of invasion >/= 10 mm or plevic lymph node metastasis. These patients may be suitable candidates for adjuvant therapy before or after surgical treatment.

Histopathological changes in the testes of prepubertal male rats after chronic administration of cocaine.

PURPOSE: This study was conducted to evaluate histopathologic findings in the testes of prepubertal male rats after long-term cocaine exposure. METHODS: At 25 days of age, male Sprague-Dawley rats were administered cocaine hydrochloride daily (15 mg/kg body weight corresponding to an average single dose for a heavy cocaine user). The treatment was continued for 100 days when all the rats were sacrificed. Morphological analysis of the testes were assessed by qualitative and quantitative histological means. RESULTS: In all the groups, a minimum of 5 to 10 representative seminiferous tubules were examined. The mean diameter of the seminiferous tubules was less in the treated group than in their respective controls (p < 0.05). The thickness of the germinal epithelium was much reduced in the cocaine-treated groups when compared with their controls (p < 0.05). The number of degenerating germ cells was greater in the treated group than in the controls. There was evidence of failure to release the mature spermatids in the treated groups. There was no evidence of sloughed Sertoli cells or germ cells in the tubular lumen or the epididymis. CONCLUSION: There were distinct histopathological changes noted after chronic administration of cocaine. These changes are characteristic of toxic effects on the testes, but the exact mechanism is not clear. Further studies are underway in our laboratory to delineate the exact mechanism of action by cocaine on the testes.

Unusual endometrial stromal cell changes mimicking metastatic carcinoma.

We report an unusual histopathologic finding in the endometrium from six premenopausal or perimenopausal patients who underwent biopsy for abnormal uterine bleeding. In each case the endometrial stromal cells focally exhibited a pronounced epithelioid appearance; some showed a cord-like trabecular pattern and others a signet-ring-like cell change. All raised the possibility of a metastatic malignancy, particularly from the breast. The epithelioid appearance of the stromal cells appears to be an unusual artifact occurring in a background of secretory change and predecidualization with nonmenstrual breakdown.