SALL1 expression in the human pituitary-adrenal/gonadal axis.

SALL1 was originally identified on the basis of its DNA sequence homology to the region-specific homeotic gene Sal, in Drosophila melanogaster, which acts as a downstream target of hedgehog/tumor growth factor-beta-like decapentaplegic signals. The SALL1 gene has been associated with the Townes-Brocks Syndrome (TBS), a disorder characterized by multiorgan dysgenesis including renal and genital malformations. In this study, SALL1 message production was evaluated in association with the tissue localization of the protein product of SALL1, p140. SALL1 protein expression was observed in various adult and fetal tissues which elaborate reproductive endocrine hormones. The p140 was localized in specific microanatomic sites of the pituitary, adrenal cortex and the placenta. In the human pituitary, SALL1 protein expression was limited to the adenohypophysis, where it colocalized to those cells producing GH and the gonadotropins, LH and FSH. SALL1 expression was also found in most of the fetal and adult adrenal cortex in addition to the trophoblastic cells of the placenta. This pattern of expression complements prior studies demonstrating p140 in testicular fetal Leydig cells, adult Leydig and Sertoli cells, and granulosa cells of the ovary. The SALL1 protein was also shown here to be highly expressed in trophoblast tumors, which overproduce sex hormones. The expression patterns of SALL1 at multiple levels of the reproductive endocrine axis and the phenotypic effects associated with TBS suggest that SALL1 may have an important role in the interaction of the pituitary-adrenal/gonadal axis during reproduction.

Hsal 1 is related to kidney and gonad development and is expressed in Wilms tumor.

Townes-Brocks syndrome (TBS) is a human genetic disorder with features including urogenital, limb, anal and cardiac malformations associated with mutations of the TBS gene, Hsal 1. To begin to understand the role of the Hsal 1 protein (p140) in both normal development and disease pathogenesis, both message and protein expression were evaluated in specific tissues associated with TBS. DNA sequence information for Hsal 1 predicts that this homeotic, Drosophila homologue (Sal) encodes a zinc-finger protein consistent with a transcription factor. mRNA for Hsal 1 was highly expressed in fetal kidney and brain, with detectable production in thymus and heart. p140 was found in fetal ureteric bud, fetal and postnatal renal tubular epithelium, and renal blastema. In the 14-week fetal testis, the Hsal 1 protein was specifically expressed in the testosterone producing Leydig cells while in adult gonads Hsal 1 was also found in both Leydig and Sertoli cells, spermatogonia of the testis, and granulosa cells of the ovary. Evaluation of Wilms tumor revealed consistently high expression of the gene product in the epithelial and blastemal components. These spatial and temporal patterns of expression for Hsal 1, and the phenotypic effects associated with TBS, suggest that Hsal 1 plays an important role in the development and functional maintenance of the kidney and gonads. Furthermore, the Hsal 1 gene product may play a part in the pathogenesis of specific neoplasms occurring in these organs in addition to its specific role in Townes-Brocks syndrome.

Atypical squamous metaplastic cells: reproducibility, outcome, and diagnostic features on ThinPrep Pap test.

BACKGROUND: Atypical squamous metaplastic (ASM) cells are associated with high-grade squamous intraepithelial lesions (HGSIL) in many cases. The reproducibility of the diagnosis and biopsy follow-up results of cases designated as ASM were studied at Women and Infants' Hospital of Rhode Island. METHODS: Of 180 patients with ASM who the authors examined from January 1, 1998 to September 30, 1998, 147 (81.7%) had subsequent biopsies. Results of the biopsies were tallied. Twenty cases were rescreened in a blinded fashion to determine intra- and interobserver agreement and to identify diagnostic features. RESULTS: Sixty-five (44.2%) cases of ASM had HGSIL on biopsy, 26 (17.7%) had low-grade squamous intraepithelial lesion, and 56 cases (38.1%) were benign. Overall individual consistency is 8 of 16 (50%), and overall agreement is 13 of 64 (20%). CONCLUSIONS: Sixty-two percent of cases designated as ASM cytologically were associated with SIL, primarily HGSIL, at biopsies. The findings underscore the importance of this subcategory of atypical squamous cells. However, poor reproducibility suggests the need for refined criteria and/or continuing education, and obtaining second opinion. Cancer (Cancer Cytopathol)

ThinPrep Pap Test promotes detection of glandular lesions of the endocervix.

The objectives of this study were to evaluate 1) the detection rate of atypical glandular cells of undetermined significance of endocervical cell type (AGUS-EC) and 2) the correlation between AGUS-EC on cytology and biopsy results using the conventional Papanicolaou (Pap) smear test vs. the ThinPrep Pap test (TPPT). Cervical-vaginal samples processed by the conventional Pap smear for 11 mo in 1996-1997 were identified, as were TPPTs collected for the same interval in 1997-1998. Biopsy results were compared after a 9-mo follow-up for both groups. There were 112 AGUS-EC cases from 82,754 conventional Pap smears (detection rate, 0.14%) compared with 58 cases from 82,252 TPPTs (detection rate, 0.07%) (P < 0.01). Biopsies were available in 72 of 112 patients from the conventional Pap smear group and 35 of 58 patients from the TPPT groups. Five dysplastic glandular lesions/ AIS were diagnosed by biopsy in the 35 patients (14.3%)from the TPPT group, compared with 2 of the 72 patients (2.8%) from the conventional Pap smear group (P < 0.05). There were no statistically significant differences between other follow-up diagnoses for the two methods. The use of TPPT resulted in fewer cases of AGUS-EC and better correlation with histology. The TPPT appears to be as sensitive as and more specific than the conventional Pap smear for detection of endocervical glandular lesions.

Secretion of activin A in recurrent epithelial ovarian carcinoma.

OBJECTIVES: Activin A is a dimeric protein, composed of two beta-A subunits, that belongs to the TGF-beta family of growth factors. Most primary epithelial ovarian tumors (96%) synthesize and secrete activin protein in vitro and preliminary studies show that serum levels of activin are frequently elevated in women with epithelial ovarian cancer. Our objectives were to expand on studies of serum activin A levels in women with epithelial ovarian cancer and to determine whether levels of activin A correlate with the clinical course of disease. METHOD: Preoperative serum activin A levels were measured in 41 patients with epithelial ovarian cancer. In addition, serum activin A levels were measured in all available postoperative samples from the subset of these patients (n = 26) who had an elevated preoperative serum activin A level. Medical record information was used to compare each patient's serum levels of activin A to the clinical course of disease. RESULTS: Seventy-two percent of the stage III and IV patients (26/36), and none (0/5) of the stage I patients, had an elevated preoperative serum activin level. In postoperative samples, activin A levels were increased with persistent or recurrent (n = 9) stage III or IV ovarian cancer. Activin A levels dropped postoperatively and remained at or below the control level in patients in remission. CONCLUSION: Serum activin A levels correlate with recurrent or persistent disease in patients with epithelial ovarian cancer.

Absence of estrogen and progesterone receptors in glassy cell carcinoma of the cervix.

OBJECTIVE: To evaluate the estrogen and progesterone receptor status of glassy cell carcinoma of the cervix to assess the possible implications of hormone replacement therapy in these patients. METHODS: The estrogen and progesterone receptor status of 13 glassy cell carcinomas was evaluated by immunohistochemistry using commercial monoclonal antibodies and a streptavidin-biotin detection system. RESULTS: No immunohistochemically detectable estrogen or progesterone receptor protein was present in tumor cells, although both receptors were identified in the adjacent normal cervical tissue. CONCLUSION: The absence of estrogen and progesterone receptors in glassy cell carcinoma suggests that this tumor would not be hormonally responsive and that these patients can be safely treated with hormone replacement therapy. Further studies are indicated to confirm this observation.

S-Phase fraction, p53, and HER-2/neu status as predictors of nodal metastasis in early vulvar cancer.

OBJECTIVE: Our aim was to determine the value of the S-phase fraction, p53, and HER-2/neu status as predictors of inguinal nodal metastasis in early vulvar cancer. METHODS: The charts of 100 consecutive patients with invasive squamous cell cancer of the vulva were reviewed and a cohort of patients with clinical stage I or II disease treated primarily with radical surgery and inguinal node dissection was identified. Within this cohort, all node-positive patients were matched with node-negative controls by depth of invasion. Tumor from the 13 node-positive patients and 26 controls was then analyzed by flow cytometry and immunohistochemistry. RESULTS: The median value of the S-phase fraction was higher in tumor from patients with inguinal nodal metastasis (median, 18.2; 25th-75th percentile: 13.9-28.3) than in node-negative patients (median, 8.9; 25th-75th percentile: 5.4-15.6) (P = 0.01). The presence of the HER-2/neu immunopositivity was also found to be associated with nodal metastasis (OR 4.05, 95% CI 1.0-16.6), but we found no evidence that DNA index or the presence of p53 immunopositivity was associated with nodal metastasis. CONCLUSION: Early vulvar cancer patients with inguinal node metastasis have a significantly higher S-phase fraction and are more likely to have HER-2/neu immunopositivity when compared to those without nodal metastasis.

Synchronous endometrioid tumors of the ovary and endometrium. A clinicopathologic study of 22 cases.

OBJECTIVE: To analyze a group of 22 patients with synchronous endometrioid tumors of the ovary and endometrium. STUDY DESIGN: A retrospective chart review was undertaken and information collected on patient age, parity, tumor grade and stage, presence of coexisting endometriosis and survival. Flow cytometry was determined from archival samples of the endometrial and ovarian tumors. RESULTS: The mean age at diagnosis was 52.8 years (range 36-71); mean parity was 1.05. With regard to the endometrial component, 68.2% were grade 1, 63.6% were stage I and, by flow cytometry, 62.5% were aneuploid. With regard to the ovarian lesions, 68.2% were grade 1, 68.2% were stage I, and 71.4% were aneuploid by flow cytometry. Twelve (54.5%) of 22 patients had pathologic evidence of coexisting endometriosis. Overall, three-year survival was 75%. All 11 patients with stage I disease at both sites were alive, without disease, at a mean follow-up of 34.9 months. CONCLUSION: Patients with synchronous endometrioid tumors of the endometrium and ovary are generally younger than reported for either endometrial adenocarcinomas or ovarian epithelial adenocarcinomas. They tend to be low grade and early stage and are frequently associated with endometriosis. Our data suggest that the survival of patients with synchronous primaries correlates with the stage of the individual tumors and that a second, synchronous primary does not adversely affect prognosis.

Tumor angiogenesis as a prognostic factor in cervical carcinoma.

Angiogenesis, the induction of new capillaries and venules, has been associated with tumor growth. Increased tumor size and new vessel growth may further the opportunity for tumor cells to enter the circulation and potentiate metastatic disease. To investigate if tumor angiogenesis could serve as a prognostic factor in cervical carcinoma, we counted microvessels (capillaries and venules) in 29 patients with squamous cell carcinoma of the cervix. Surgical specimens were stained for endothelial cells specifically with Factor VIII to identify all vessels. The microvessels were counted by light microscopy (per 200 x field) in tumor sections with the highest population of microvessels. This was performed by two investigators without knowledge of patient outcome or extent of disease. Microvessel counts in patients with squamous cell carcinoma were significantly different from those of control subjects: 56 +/- 28.9 and 16.3 +/- 3.3 (P = 0.013). There was no correlation between microvessel count and node status, parametrial involvement, depth of invasion, or gross disease. Microvessel count was significantly correlated with vascular space involvement (P = 0.017). Four patients who developed recurrent disease within 1 year had high microvessel counts and yet were node negative and VSI negative at surgery. As shown by Folkman in breast cancer, angiogenesis may also be an independent predictor for recurrent disease in squamous cell carcinoma of the cervix. Microvessel counts could be of prognostic value in patients who do not have other risk factors for disease recurrence.

Sperm electron microscopy for the evaluation of in vitro fertilization failures.

When performing IVF, the clinician is frequently confronted with the failure of fertilization. When the standard parameters to evaluate the male factor are "within normal limits," the conclusion is often made that the lack of fertilization is most likely due to "poor egg quality." These two cases demonstrate the fallacy of this approach and support a more rigorous evaluation of the male factor. Ultrastructural analysis of sperm is underutilized and, as demonstrated by these two cases, can play an essential role in this evaluation process.

Stromal amyloid in pheochromocytomas.

Amyloid has been documented in the stroma of a number of neuroendocrine tumors. It is usually associated with elaboration of a polypeptide hormone product. Twenty-three adrenal pheochromocytomas occurring in 18 patients were graded for the extent of amyloid deposits on the hematoxylin-eosin-stained slides, confirmed to be amyloid by Congo red stain and polarization microscopy. Fourteen of 20 cases (70%) showed evidence of stromal amyloid; in two thirds of these cases, it was considered abundant. The awareness of amyloid deposits in approximately 70% of pheochromocytomas is important for surgical pathologists, as this occurrence has not been thought to be common in these tumors and might be a source of diagnostic difficulty.

Triplet tubal pregnancy treated by outpatient laparoscopic salpingostomy.

To our knowledge, this represents the first case of a laparoscopically treated triplet EP and the first time that a double EP in the same tube was treated conservatively (with preservation of the tube). Multiple EPs may be more common than currently thought, and our report offers an alternative explanation for at least some cases of persistent EP after conservative surgical therapy. Finally, given the substantial cost savings and reduced postoperative recovery time associated with operative laparoscopy, when the patient is stable and the surgeon experienced, the laparoscopic approach should be tried, regardless of the number of EPs or their size.

Immature teratoma of the ovary.

Twenty-five cases of patients with pure immature teratoma of the ovary, accrued from the Connecticut Tumor Registry from 1969 to 1984, were reviewed. Two patients had grade 1 tumors, twelve had grade 2 tumors, and eleven had grade 3 tumors. The majority of patients (23) were stage I at the time of initial surgery. Twenty-one of the twenty-three patients were treated with some form of unilateral adnexal surgery with or without adjuvant combination chemotherapy (VAC). Two of the twenty-three patients were treated with total abdominal hysterectomy/bilateral salpingo-oophorectomy (TAH/BSO) with the addition of either VAC chemotherapy or radiation therapy. Recurrence occurred in two patients, both of whom had grade 3 tumors and were subsequently treated with surgical resection plus VAC chemotherapy. One patient, who recurred after initial therapy with unilateral salpingo-oophorectomy (USO) plus VAC chemotherapy, was successfully treated with surgical resection and further chemotherapy. Two patients were stage III at the time of initial surgery, one of whom was treated with USO plus adjuvant combination chemotherapy and radio-therapy. The other patient was treated with TAH/BSO plus VAC chemotherapy. In our series, no patient died from immature teratoma (one patient died from advanced breast carcinoma). It is reasonable to withhold chemotherapy from patients with stage I, grade 1 and 2, immature teratoma which may be treated initially with conservative surgery. The risk of recurrence in patients with grade 3 tumors warrants the addition of further chemotherapy.

Mercury-induced renal autoimmunity in the MAXX rat.

Inbred Brown Norway (BN) rats treated with mercuric chloride develop autoantibodies to renal basement membranes and an immunologically mediated membranous glomerulonephritis. To date, this experimental rat model of chemically induced autoimmunity has been obtained only in the BN strain, whereas rats from 17 other strains were found to be resistant. This is a disadvantage for mechanistic studies, especially since BN rats have poor fertility. In the present paper we report that the same model can be obtained in another inbred strain of rats, the MAXX, which after exposure to mercury develop a glomerulonephritis characterized by the production of autoantibodies to renal basement membranes. The kinetics of the autoimmune response observed in MAXX rats, as well as the immunohistopathology, histopathology, and proteinuria, are similar to those previously described in BN rats. In addition, the MAXX strain is endowed with excellent fertility. Therefore, both rat strains can be used for comparative studies of the mechanisms of mercury-induced autoimmunity.

Axonal degeneration/necrosis: a possible ultrastructural marker for Crohn's disease.

Axonal degeneration/necrosis has previously been demonstrated in the small bowel autonomic nerve plexus of patients with Crohn's disease. It was suggested that this feature might be helpful in the differential diagnosis of Crohn's disease from other conditions, specifically ulcerative colitis. We performed a blind prospective study and examined tissue from 34 colonic specimens, in 15 patients, by electron microscopy. Cases of Crohn's disease (3 cases), ulcerative colitis (5 cases), and other controls [adenocarcinoma (4 cases), familial polyposis (1 case), peritonitis (1 case), and radiation colitis (1 case)] were evaluated for inflammation and axonal changes. It was found that only the cases of Crohn's disease showed extensive, seemingly independent, severe axonal changes that were unassociated with other markers of inflammation. This feature may be useful in the differential diagnosis between Crohn's disease and ulcerative colitis.

Enzymatic formation of prostaglandin D2 by rat basophilic leukemia cells and normal rat mast cells.

It has been shown that the major cyclooxygenase product in rat basophilic leukemia (RBL-1) cells and in normal rat mast cells is prostaglandin D2. In RBL-1 cells, prostaglandin D2 is isomerase activity was found in the 150 000 X g microsomal pellet as well as the supernatant fraction. Incubation of RBL-1 microsomes with arachidonic acid without cofactors yielded 17.5 +/- 2% prostaglandin E2 and 9.1 +/- 1.4% prostaglandin D2. The cyclooxygenase activity was enhanced (25%) by epinephrine and the addition of reduced glutathione led to a marked increase in prostaglandin D2 synthesis (3-fold). Incubations with arachidonic acid, glutathione and epinephrine gave the maximum conversion to prostaglandin D2, yielding 7 +/- 0.4% prostaglandin E2 and 35.6 +/- 3.5% prostaglandin D2. Incubations with [14C]prostaglandin H2 to bypass cyclooxygenase confirmed the presence and glutathione dependence of the prostaglandin D2 isomerase in the microsomal fraction and also revealed the presence of the same enzyme in the 150 000 X g supernant. In contrast to RBL-1 cells, incubations of microsomes and supernatant from normal rat mast cells with [14C]-arachidonic acid and [14C]prostaglandin H2 localized the prostaglandin D2 isomerase activity in the soluble fraction. Similar to the enzyme in the RBL-1 cells, the mast cell enzyme was glutathione dependent.