The effects of frozen tissue storage conditions on the integrity of RNA and protein.

Unfixed tissue specimens most frequently are stored for long term research uses at either -80° C or in vapor phase liquid nitrogen (VPLN). There is little information concerning the effects such long term storage on tissue RNA or protein available for extraction. Aliquots of 49 specimens were stored for 5-12 years at -80° C or in VPLN. Twelve additional paired specimens were stored for 1 year under identical conditions. RNA was isolated from all tissues and assessed for RNA yield, total RNA integrity and mRNA integrity. Protein stability was analyzed by surface-enhanced or matrix-assisted laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS, MALDI-TOF-MS) and nano-liquid chromatography electrospray ionization tandem mass spectrometry (nLC-ESI-MS/MS). RNA yield and total RNA integrity showed significantly better results for -80° C storage compared to VPLN storage; the transcripts that were preferentially degraded during VPLN storage were these involved in antigen presentation and processing. No consistent differences were found in the SELDI-TOF-MS, MALDI-TOF-MS or nLC-ESI-MS/MS analyses of specimens stored for more than 8 years at -80° C compared to those stored in VPLN. Long term storage of human research tissues at -80° C provides at least the same quality of RNA and protein as storage in VPLN.

Establishing a tumour bank: banking, informatics and ethics.

The six divisions of the Cooperative Human Tissue Network in the USA bank and distribute tens of thousands of tissue specimens to researchers annually. Major operational concerns include: maintaining tissue integrity, managing informatics, and protecting patient confidentiality. Increasing molecular genetics testing is also resulting in an increased demand for high-quality nucleic acids.

Benign glandular and squamous metaplastic-like cells seen in vaginal Pap smears of post hysterectomy patients: incidence and patient profile.

We have observed benign glandular cells and squamous metaplastic-like cells in vaginal Pap smears of post hysterectomy patients (PHP). Vaginal Pap smears from 1,547 PHP were retrieved. In 2% of these smears (Group A) glandular cells were observed, with the majority of the smears revealing squamous metaplastic-like cells (47%). Mucinous endocervical columnar-like cells were seen in 9% of the cases, glandular cells not resembling endocervical cells in 13%, and a combination of the former two categories in 31%. Group A patients were compared with other PHP without these cells in their vaginal smears (Group B). Several clinical and surgical parameters were evaluated. A distinctive clinical profile was not identified for either group of patients (A or B). Of patients in group A 49.8% had a history of a previous gynecologic malignancy (Group B: 19%). Based on our study, we postulate that in the absence of a clinically identifiable source of these cells, the most likely source of origin is probably vaginal adenosis not associated with DES exposure in utero or a metaplastic phenomenon perhaps related to therapy. These cells do not seem to be related to imminent neoplasia or dysplasia.

Ovarian smooth muscle metaplasia: an uncommon and possibly underrecognized entity.

Smooth muscle differentiation in the ovary is rare, and its histopathologic spectrum, including ovarian smooth muscle metaplasia (SMM), has not been well described. The clinicopathologic findings in 48 ovaries with SMM from 40 women are reported. The average age of women with ovarian SMM was 55.6 years (range, 34 to 86 years). Foci of SMM were semiquantitatively characterized as 1+ in 46% (1 to 3 foci), 2+ in 37% (4 to 6 foci), and 3+ in 17% (> 6 foci). SMM was bilateral in 8 (23%) of the 35 patients who had bilateral oophorectomies. SMM was intimately associated with another ovarian process in 28 (58%) cases, including ovarian cysts (11), endometriosis (3), granulosa cell tumors (3), extensive stromal luteinization (1), ovarian fibroma (1), adhesions (1), and folliculogenesis (8). Ovaries with 2+ to 3+ SMM were associated with another ovarian lesion significantly more often than those with 1+ SMM (p < 0.01). Most women with ovarian SMM (86%) also had uterine leiomyomas. Significant endometrial pathology was present in 13 (37%) of 35 simultaneously removed uteri. In conclusion, SMM occurs most often in perimenopausal or postmenopausal women, most of whom also have uterine leiomyomas. Ovarian SMM is usually confined to a few microscopic fields, is bilateral in < 25% of patients, and is often associated with other ovarian lesions.

Ovarian leiomyomas: clinicopathologic features in fifteen cases.

Ovarian leiomyomas are uncommon and have not been systematically studied. The clinical and histopathologic features of 15 ovarian leiomyomas were evaluated, including their clinical presentation, size, cellularity, mitotic index, presence of degeneration, and hyalinization. The mean age of women with ovarian leiomyomas was 45.8 years. The presenting sign was an adnexal mass in 7 (47%), uterine leiomyomas in 4 (26%), contralateral adnexal mass in 2 (13%), and other signs in 2 (13%) women. Thirteen leiomyomas had mitotic indices < 1/10 high-power-fields (HPFs), including one cellular leiomyoma, and 2 had mitotic indices between 1 and 2/10 HPFs. The cellular leiomyoma and those with a mitotic index of > or = 1/10 HPFs had an average size of 3.4 cm. Two leiomyomas exhibited central degeneration, and hyaline change was present in 5 leiomyomas, 4 of which were at least 4 cm in size. The majority of ovarian leiomyomas (78%) were associated with uterine leiomyomas; hyperplastic endometrial polyps were present in 2 cases. Follow-up (mean, 9.6 months) was available in 8 cases, and all had no evidence of recurrence. In conclusion, ovarian leiomyomas exhibit a spectrum of features that are similar to uterine leiomyomas. Although the prognosis is presumed to be good, the short follow-up period of this series precluded definitive evaluation of their natural history.

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.

Retained trophoblastic tissue in fallopian tubes: a consequence of unsuspected ectopic pregnancies.

Five patients had retained trophoblastic tissue within the Fallopian tubes, which suggested remote ectopic pregnancies. Four remote ectopic pregnancies were identified in patients after delivery. One patient underwent salpingo-oophorectomy after the identification of a small ovarian mass and 1.0 cm "necrotic area" in the fallopian tube at cesarean section. The other three patients underwent tubal ligation, and one was noted to have a 1.0 cm "calcified" nodule in the distal fallopian tube. The fifth patient underwent laparotomy and was found to have an acute ectopic pregnancy in one fallopian tube and a clinically unsuspected 3.0 cm mass in the other, which proved to be an ectopic pregnancy with ghost outlines of chorionic villi and trophoblast. Histopathologically, all five patients showed foci of viable-appearing intermediate trophoblast and surrounding abundant eosinophilic hyalinized material in the fallopian tubes; four patients also had hyalinized ghost outlines of chorionic villi. None of the mothers had a history of previous ectopic pregnancy. The natural history of clinically unsuspected ectopic tubal pregnancies is not well understood, but these cases illustrate that trophoblast may persist in fallopian tubes and potentially result in clinical confusion as well as tubal pathology.

Tumors of the uterine isthmus: clinicopathologic features and immunohistochemical characterization of p53 expression and hormone receptors.

Five (3%) of 161 endometrial cancers treated surgically between 1988 and 1992 were classified as primary isthmic tumors, and their clinicopathologic features, p53 expression, and hormone receptor status were evaluated. Three were endometrioid adenocarcinomas with squamous differentiation, one was a mixed serous and clear cell carcinoma, and one was a malignant müllerian mixed tumor; all five were high grade, invaded the entire thickness of the myometrium, and exhibited lymphatic space invasion. Four of the five patients had extrauterine metastases identified at the time of hysterectomy. All five patients died due to progressive disease with the survival time ranging from 1 to 23 months. Abnormal p53 gene expression was identified immunohistochemically in three of the five isthmic tumors. Weak positivity for estrogen and progesterone receptors was demonstrated in one case, with the remaining four being negative. Tumors arising in and confined to the uterine isthmus are unusual and, in our series, were uniformly aggressive with an unfavorable prognosis. The histopathologic features and biologic behavior of the isthmic tumors appeared similar to those of other high-grade endometrial cancers arising in the uterine corpus.

Ectopic pregnancy. A recent five-year study and review of the last 50 years' literature.

OBJECTIVE: To compare the past and current clinical and pathologic factors that are responsible for ectopic pregnancy (EP). STUDY DESIGN: We performed a retrospective chart and histopathologic specimen review of 740 cases of EP during a recent, five-year period. Products of conception, as well as 240 cases of concomitant endometrial biopsies, were examined. RESULTS: The mean age at presentation, proportion of patients with a history of pelvic inflammatory disease, voluntary interruption of pregnancy (VIP), previous surgery and laterality (right side) of the tubal EP were similar to rates reported in previous studies. Ninety-four percent of the EPs were tubal, and 90% of the tubes exhibited some pathologic changes, including chronic salpingitis (49.5%) and follicular salpingitis (10%), among others. Gestational endometrial patterns were seen in 44% of cases. The corpus luteum of pregnancy was contralateral in 30% of cases in which an ovarian biopsy was performed. CONCLUSION: The factors classically associated with EP remained similar over a period of about 50 years, although the association with VIP still appears to be controversial. Endometrial curettage alone cannot be used to exclude an EP. Ovum transmigration may play a role in the genesis of EP.

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.

Placenta accreta: mild cases diagnosed by placental examination.

We describe 36 placentas in which microscopic foci of myometrial tissue were adherent to the basal plate with deficient intervening decidua, consistent with a mild or focal form of placenta accreta. In only four cases was a diagnosis of placenta accreta considered clinically, and none of the cases resulted in hysterectomy. Twenty-one of the mothers underwent cesarean section for the current pregnancy, with 10 of the mothers having a history of previous cesarean section. The placenta failed to separate spontaneously in eight of the 15 vaginal deliveries necessitating manual removal of the placenta. Additional maternal factors included multiparity in 33, placenta previa in two, leiomyomas in two, and previous spontaneous abortion or voluntary interruption of pregnancy in 23. Four cases were complicated by retained placental fragments and three by postpartum hemorrhage. We conclude that these milder cases of placenta accreta are frequently associated with previous uterine operations and multiparity, and although not uncommon, are frequently not clinically suspected. Placental examination is useful in making the diagnosis of placenta accreta in cases not requiring hysterectomy, particularly if the basal plate is well sampled.