The histone demethylase KDM4B regulates peritoneal seeding of ovarian cancer.

Epithelial ovarian cancer (EOC) has poor prognosis and rapid recurrence because of widespread dissemination of peritoneal metastases at diagnosis. Multiple pathways contribute to the aggressiveness of ovarian cancer, including hypoxic signaling mechanisms. In this study, we have determined that the hypoxia-inducible histone demethylase KDM4B is expressed in ∼60% of EOC tumors assayed, including primary and matched metastatic tumors. Expression of KDM4B in tumors is positively correlated with expression of the tumor hypoxia marker CA-IX, and is robustly induced in EOC cell lines exposed to hypoxia. KDM4B regulates expression of metastatic genes and pathways, and loss of KDM4B increases H3K9 trimethylation at the promoters of target genes like LOXL2, LCN2 and PDGFB. Suppressing KDM4B inhibits ovarian cancer cell invasion, migration and spheroid formation in vitro. KDM4B also regulates seeding and growth of peritoneal tumors in vivo, where its expression corresponds to hypoxic regions. This is the first demonstration that a Jumonji-domain histone demethylase regulates cellular processes required for peritoneal dissemination of cancer cells, one of the predominant factors affecting prognosis of EOC. The pathways regulated by KDM4B may present novel opportunities to develop combinatorial therapies to improve existing therapies for EOC patients.

Comparison of immunohistochemistry by automated cellular imaging system (ACIS) versus fluorescence in-situ hybridization in the evaluation of HER-2/neu expression in primary breast carcinoma.

AIMS: Immunohistochemistry (IHC) and fluorescence in-situ hybridization (FISH) are both commonly used assays for evaluation of HER-2/neu status in breast cancer. However, there is still no consensus on which method is most predictive of patient response to Herceptin. Recently, the automated cellular imaging system (ACIS) has been shown to improve the accuracy and reproducibility in scoring IHC. Our aim was to compare the results of HER-2/neu expression and gene amplification in the same patients by IHC using the ACIS system and by FISH. METHODS AND RESULTS: Two hundred and forty-seven breast cancer cases were studied. The concordance rate between IHC-ACIS (> or = 2.2) and FISH (> or = 2.0) was 94%. Fifteen patients were discordant; three had borderline FISH values and three had borderline IHC values. The other nine discordant cases consisted of five IHC-ACIS+, FISH- and six IHC-ACIS-, FISH+. HER-2/neu overexpression was more common in tumours that were high-grade, aneuploid, progesterone receptor and bcl-2 negative, with MIB-1 > 10%. CONCLUSION: HER-2/neu assessment by the ACIS is reliable, rapid and inexpensive, and correlates highly with results obtained by FISH.

Breast conservation therapy with tumor bed irradiation alone in a selected group of patients with stage I breast cancer.

Radiotherapy after breast-conserving surgery increases local control. We tested the feasibility of limited surgery with tumor bed irradiation only with 192Ir in a selected group of patients with stage I breast cancer. Twenty-five breasts in 24 women more than 60 years old with low- or intermediate-grade stage I tumors were treated with placement of interstitial catheters at the time of lumpectomy and axillary node dissection. This procedure was followed by after-loading with low-dose 192Ir to deliver 20-25 Gy to the tumor bed over 24-48 hours. There were neither local recurrences in the breast nor distant recurrences at a median follow-up of 47 months (range 25-90 months). Cosmetic appearance ranged from very good to excellent. There were no long-term complications. It is feasible to treat a select group of patients with tumor bed irradiation, using relatively low doses of interstitial irradiation, with excellent local control and no significant morbidity.

CD44s expression is reduced in endometriotic lesions compared to eutopic endometrium in women with endometriosis.

Immunohistochemical expression of the standard form of CD44 (CD44s) was examined in archival formalin-fixed endometriotic and matching eutopic endometrial tissue obtained from 25 patients in proliferative (N = 16) and secretory (N = 9) stages of the cycle. CD44s was expressed in most eutopic endometria and endometriotic tissue. Its expression was significantly higher in secretory than in proliferative phase endometrium. It was low but detectable in 13 of 16 proliferative phase biopsies. The majority of these endometria exhibited both glandular and stromal staining (63%). In the secretory phase, glandular cells exhibited a significantly greater intensity of staining compared to stromal cells. In endometriotic tissue, stromal cell CD44s expression did not differ between tissue types in either stage of the cycle. In contrast, glandular expression in endometriotic tissue during the secretory phase was reduced (p < 0.05) compared to eutopic endometrium. It was absent in 66% of cases and reduced in the remaining cases. Our results indicate a correlation between CD44s expression and secretory differentiation of endometrial glands in the cycle, suggesting hormonal regulation of its expression. This cyclic pattern of CD44s expression was lost in corresponding endometriotic tissue. Reduced expression of CD44s in endometriotic tissue may provide insight into the pathophysiology of endometriosis.

Langerhans cell granulomatosis manifested as pigmented villonodular synovitis.

We report an unusual case of Langerhans cell granulomatosis (LCG) manifested as a villous synovial proliferation in a 38-year-old female jogger. One year after the onset of joint symptoms, she had a classical LCG presentation with skin and visceral lymph node involvement. Review of the literature revealed only one case of synovial shoulder joint tenosynovitis associated with LCG in a middle-aged woman. Ours is the first reported case presenting clinically in the synovium of the hip joint as pigmented villonodular synovitis. Histiocytic/dendritic proliferations involving the synovial tissues are not uncommon. These lesions as well as the rare multicentric reticulohistiocytosis (MRH), a systemic monocytoid/histiocytic disorder with multinucleated giant cells, polyarthritis, and papulonodular skin lesions, should be considered in the differential diagnosis. Clinical and pathologic features will distinguish LCG from MRH.

Tumor necrosis factor-alpha response to infection with Chlamydia trachomatis in human fallopian tube organ culture.

OBJECTIVE: Our purpose was to determine whether tumor necrosis factor-alpha is produced in response to infection with Chlamydia trachomatis in the fallopian tube. STUDY DESIGN: Fallopian tubes were harvested at the time of abdominal hysterectomy and processed by standard tissue culture techniques. Tubal segments were inoculated with Chlamydia trachomatis serotype E/UW-5/CX. At 48 hours of incubation supernatant fluid was assayed for tumor necrosis factor-alpha. Tubal segments were stained for chlamydial inclusions and tumor necrosis factor-alpha by use of immunohistochemical techniques. RESULTS: Mean tumor necrosis factor-alpha levels for infected segments were 92.1 +/- 21.3 pg/ml (mean +/- SEM) and for control segments were 61.9 +/- 13.9 pg/ml (p = 0.03 by paired t test). Tumor necrosis factor-alpha was predominantly localized in the tubal epithelium. CONCLUSIONS: Tumor necrosis factor-alpha is produced in response to chlamydial infection by the human fallopian tube. It is an important proinflammatory cytokine and may promote the production of other cytokines and immune-mediated damage of the fallopian tube.

Multicentric bilateral renal cell carcinomas and a vascular leiomyoma in a child.

We report a unique case of multicentric bilateral renal cell carcinomas and a simultaneous large renal vascular leiomyoma in an 11-year-old child with sickle cell anemia. The patient presented with several episodes of massive hematuria. Abdominal sonography and computed tomography demonstrated bilateral renal neoplasms and the patient was clinically thought to have bilateral Wilms' tumor. An initial biopsy of the lower pole of right kidney revealed a renal cell carcinoma. Accordingly, bilateral renal angiography followed by right total nephrectomy and left upper pole partial nephrectomy were performed. Pathologic studies showed multicentric, bilateral renal cell carcinomas (two in the right kidney and one in the left kidney), of clear, granular, and oncocytic cell types. A simultaneous large vascular leiomyoma was also present in the right kidney. The smooth muscle nature of the leiomyoma was determined by light microscopy, immunohistology, and electron microscopy. The diagnostic difficulties in distinguishing them from other renal tumors are discussed.

Lipohyperplasia of the ileocecal valve.

Submucosal lipohyperplasia of the ileocecal valve (ICV) is reportedly a rarely diagnosed lesion of uncertain significance. Eight cases of ICV lipohyperplasia diagnosed in surgical specimens (seven resections, one biopsy) are reviewed: three cases were associated with right lower abdominal quadrant pain and ICV mass on barium enema or operative examination, two were associated with ICV mucosal acute inflammation and necrosis, and three were incidental in resections for cecal, appendiceal, and sigmoid neoplasia. To evaluate the frequency of ICV lipohyperplasia and any associated processes, a series of 51 autopsies was studied. Regarding lipohyperplasia in these valves, 10 (19.6%) were determined to have none, 14 (27.5%) were mild, 20 (39.2%) were moderate, and 7 (13.7%) were marked cases. Degree of lipohyperplasia correlated statistically with degree of cardiac right ventricular fatty infiltration (p = 0.0001), pancreatic fatty infiltration (p = 0.0314), and greater body weight of patient (p = 0.0009). No definite correlation was demonstrated with left ventricular, adrenal, or lymph node fatty infiltration, or with hepatic fatty change, body height, age of patient, or blood glucose. Various gastrointestinal symptoms and lesions accompanied lipohyperplasia, but no definite causal relationship was identified, except for one case of marked lipohyperplasia associated with marked mucosal necrosis and acute inflammation of ICV. In conclusion, ICV lipohyperplasia is a common finding that occasionally may be associated with clinical symptoms and other valve pathology. It correlates to some extent with right ventricular and pancreatic fatty infiltration and with greater body weight.

Saccharomyces cerevisiae pneumonia in a patient with acquired immune deficiency syndrome.

The clinical course of a patient with a polymicrobial pneumonia that included Saccharomyces cerevisiae infection is described. S. cerevisiae was recovered from autopsy cultures of the lungs, spleen, oral mucosa, and small intestine, and organisms morphologically consistent with S. cerevisiae were visualized in histologic sections of the lung. The role of this organism as a human pathogen is reviewed.

Epidermal growth factor binding in rat uterus during the peri-implantation period.

The profile of epidermal growth factor (EGF) binding to uterine membranes of rats on Day 1 through Day 7 of pregnancy was studied. The binding was lowest on Day 1 and increased gradually through the pre- and postimplantation periods. Binding affinity of the Day 7 uterine membranes was considerably higher than that of the Day 1. Apparent affinity constants (Ka) of Day 1 and Day 7 membranes were 0.29 X 10(-8) M and 1.03 X 10(-8) M respectively. To our knowledge, this is the first report of the modulation of EGF binding to uterine membranes by progesterone-estrogen interaction during early pregnancy.

Further evidence for role of leukotrienes as mediators of decidualization in the rat.

Inhibitors of leukotrienes were utilized to investigate the role of leukotrienes (LTs) in the induction of decidualization in the rat. Alzet osmotic minipumps, filled with either FPL 55712 (FPL, a specific antagonist of peptidoleukotrienes), nordihydroguaiaretic acid (NDGA, an inhibitor of LT synthesis) or in combination with leukotriene C4 (LTC4) and/or prostaglandin E2 (PGE2), were instilled at the ovarian end of uterine horns of day 5 pseudopregnant rats. Intraluminal infusion of FPL or NDGA, for 4 days, induced a dose dependent decrease in the uterine wet weights when compared to that induced by the infusion of their corresponding vehicles (1 microliter/h). Furthermore, simultaneous infusion of LTC4 (10 ng/h) with different doses of FPL (1, 0.5, or 0.25 microgram/h) produced an increase in uterine weights as compared to that produced by FPL alone. Maximum response, however, was noted when LTC4 (10 ng/h) was infused with FPL at a rate of 0.5 microgram/h. The infusion of LTC4 (10 ng/h) or PGE2 (1 microgram/h) with NDGA, at 1 and 5 micrograms/h, could not overcome its inhibitory effect on decidualization. On the contrary, a combination of LTC4 (10 ng/h) and PGE2 (1 microgram/h) along with NDGA (5 micrograms/h) significantly increased the uterine weight to a level that was comparable to that induced by the infusion of the vehicle. To determine if the synthesis of PGs and LTs was inhibited by NDGA, one uterine horn was infused with NDGA (5 micrograms/h) and the other horn with the vehicle. The intrauterine infusion of NDGA for 24 h inhibited the release of PGE2, PGF2 alpha, LTC4 and LTB4 as compared to those released by the vehicle-infused horns. These data suggest that both PGs and LTs are required for the induction and progression of decidualization.

Release of prostaglandins and leukotrienes from the rat uterus is an early estrogenic response.

The early estrogenic responses are considered to be involved in inducing embryo implantation in a progesterone (P4)-primed uterus. Because of their involvement in the process of implantation and decidualization, prostaglandins (PGs) and leukotrienes (LTs) could be the mediators of early estrogenic responses in a P4-primed uterus. Therefore, temporal effects of estrogen on the production and/or release of PGF2, PGF2 alpha, LTB4 and LTC4 by the P4-primed uterus of hypophysectomized rats were examined. Hypophysectomized mature female rats were injected for 4 days with P4 (2 mg/rat, s.c.) or with P4 plus a single injection of estradiol-17 beta (E2) (100 ng or 200 ng/rat, i.v.) on the last day of P4 treatment. In one set of experiments, animals were killed at 0.5, 2, 4, 8, 12 and 30th after the last steroid treatment. The production of PGs and Lts by uterine homogenates was measured by radioimmunoassays (RIAs). The production of PGE2 and PGF2 alpha in P4-treated animals showed peaks at 2, 6 and 12h. The superimposition of E2 on P4 treatment induced a higher production rate of PGE2 and PGF2 alpha at 0.5h and abolished the peaks induced by P4 at 2h, but not the peaks at 6 or 12h. Irrespective of the kind of steroid hormonal treatments, uterine production of LTs showed a rapid decline between 6 and 8h followed by a sharp rise at 12h. The superimposition of E2 on P4-treatment again increased the production rates of LTB4 and LTC4 at early hours, i.e. at 0.5 and 2h, respectively, as compared to P4 treatment only.

Decidualization in the rat: role of leukotrienes and prostaglandins.

The role of prostaglandins (PGs) and leukotrienes (LTs) in the induction of decidualization in the rat uterus was investigated. In the hypophysectomized progesterone (P4) primed rat, intraluminal infusion for four days by osmotic minipump, of PGE2 (1 ug/h), LTC4 (10 ng/h) or 0.15M saline (1 ul/h) significantly elevated uterine weight when compared to the noninfused horn: all were equally effective. In contrast, simultaneous infusion of PGE2 and LTC4 produced an increase in uterine weight which was markedly higher than any other conditions and the reaction was elicited along the entire length of the uterine horn. Infusion of PGE2, LTC4, a combination of the two or vehicle, into one uterine horn of day-5 pseudopregnant rats elicited a huge decidual response. Infusion of indomethacin, an inhibitor of PG synthesis, FPL 55712 (FPL), an antagonist of LTs, or a combination of these inhibitors evoked a minimal decidual response. In addition, FPL infused along with PGE2 or LTC4 markedly reduced the response that could be induced by these arachidonate metabolites alone. Furthermore, infusion of indomethacin along with LTC4 resulted in a far smaller response than that obtained with LTC4 alone. These results are interpreted to indicate that there is an interaction between LTs and PGs in the induction of the uterine decidual response.

Implication of prostaglandin E2 in soluble factor-mediated immune suppression by murine decidual cells.

Cells from the uteri of pregnant mice mediate profound nonantigen-specific and nonmajor histocompatibility complex-restricted immune suppression in vitro. In part, those cells accomplish suppression by releasing soluble suppressor factors. The purpose of the present study was to initiate identification of uterine cell suppressor factors. Immune suppression was assayed by the effect of decidual cells or in vitro generated supernatants of decidual cells on mixed lymphocyte reactions (MLR). The following findings support the designation of prostaglandin E2 (PGE2) as a primary suppressor molecule originating with decidual cells: Suppression mediated by supernatants of decidual cells was relieved by removal of lipids but not proteins; indomethacin, an inhibitor of prostaglandin synthesis, produced partial relief of suppression mediated by uterine cells and totally inhibited soluble suppressor factor generation by those cells; decidual cells produced high levels of both PGE2 and PGF2a; the addition of exogenous PGE2 at levels comparable to those found in the decidual cell supernatants restored suppression by decidual cells and their supernatants whereas the addition of PGF2a had no effect; inhibition of the lipoxygenase pathway of arachidonate metabolism had no effect on cell or supernatant mediated suppression; nonspecific suppressor mechanisms, such as arginine depletion and peroxide generation, were excluded as possible mediators of MLR suppression by decidual cells and their supernatants. Fractionation of decidual cells revealed at least three indomethacin-sensitive cell types: small, lymphocyte-like cells, macrophages, and a third population of large decidual cells that was not identified by specific markers.(ABSTRACT TRUNCATED AT 250 WORDS)

Partial characterization of uterine cells responsible for suppression of murine maternal anti-fetal immune responses.

Several studies have demonstrated that uterine cells are capable of suppressing in vitro immune responses in a nonspecific manner. Two types of cells have been implicated as responsible for suppression, cells with the features of macrophages and a small lymphocyte-like cell. In the present study the maternal anti-paternal mixed lymphocyte reaction was used to investigate further the characteristics of uterine suppressor cells. Three distinct suppressor cell populations were identified: highly suppressive macrophages, small lymphocyte-like cells, and a morphologically heterogeneous third population of highly suppressive cells. The data from this study suggest that pregnant murine uteri contain a variety of cells capable of discouraging lymphocyte proliferation in vitro.