Simvastatin interferes with cancer 'stem-cell' plasticity reducing metastasis in ovarian cancer.

Cell plasticity of 'stem-like' cancer-initiating cells (CICs) is a hallmark of cancer, allowing metastasis and cancer progression. Here, we studied whether simvastatin, a lipophilic statin, could impair the metastatic potential of CICs in high-grade serous ovarian cancer (HGS-ovC), the most lethal among the gynecologic malignancies. qPCR, immunoblotting and immunohistochemistry were used to assess simvastatin effects on proteins involved in stemness and epithelial-mesenchymal cell plasticity (EMT). Its effects on tumor growth and metastasis were evaluated using different models (e.g., spheroid formation and migration assays, matrigel invasion assays, 3D-mesomimetic models and cancer xenografts). We explored also the clinical benefit of statins by comparing survival outcomes among statin users vs non-users. Herein, we demonstrated that simvastatin modifies the stemness and EMT marker expression patterns (both in mRNA and protein levels) and severely impairs the spheroid assembly of CICs. Consequently, CICs become less metastatic in 3D-mesomimetic models and show fewer ascites/tumor burden in HGS-ovC xenografts. The principal mechanism behind statin-mediated effects involves the inactivation of the Hippo/YAP/RhoA pathway in a mevalonate synthesis-dependent manner. From a clinical perspective, statin users seem to experience better survival and quality of life when compared with non-users. Considering the high cost and the low response rates obtained with many of the current therapies, the use of orally or intraperitoneally administered simvastatin offers a cost/effective and safe alternative to treat and potentially prevent recurrent HGS-ovCs.

Lipophilic but not hydrophilic statins selectively induce cell death in gynaecological cancers expressing high levels of HMGCoA reductase.

Recent reports have suggested that statins induce cell death in certain epithelial cancers and that patients taking statins to reduce cholesterol levels possess lower cancer incidence. However, little is known about the mechanisms of action of different statins or the effects of these statins in gynaecological malignancies. The apoptotic potential of two lipophilic statins (lovastatin and simvastatin) and one hydrophilic statin (pravastatin) was assessed in cancer cell lines (ovarian, endometrial and cervical) and primary cultured cancerous and normal tissues. Cell viability was studied by MTS assays and apoptosis was confirmed by Western blotting of PARP and flow cytometry. The expressions of key apoptotic cascade proteins were analysed. Our results demonstrate that both lovastatin and simvastatin, but not pravastatin, selectively induced cell death in dose- and time-dependent manner in ovarian, endometrial and cervical cancers. Little or no toxicity was observed with any statin on normal cells. Lipophilic statins induced activation of caspase-8 and -9; BID cleavage, cytochrome C release and PARP cleavage. Statin-sensitive cancers expressed high levels of HMG-CoA reductase compared with resistant cultures. The effect of lipophilic statins was dependent on inhibition of enzymatic activity of HMG-CoA reductase since mevalonate pre-incubation almost completely abrogated the apoptotic effect. Moreover, the apoptotic effect involved the inhibition of synthesis of geranylgeranyl pyrophosphate rather than farnesyl pyrophosphate. In conclusion, lipophilic but not hydrophilic statins induce cell death through activation of extrinsic and intrinsic apoptotic cascades in cancerous cells from the human female genital tract, which express high levels of HMG-CoA reductase. These results promote further investigation in the use of lipophilic statins as anticancer agents in gynaecological malignancies.

Progesterone pre-treatment potentiates EGF pathway signaling in the breast cancer cell line ZR-75.

Progesterone in hormone replacement therapy (HRT) preparations increases, while hysterectomy greatly reduces, the incidence of breast cancer. Cross-talk between the progesterone and growth factor signaling pathways occurs at multiple levels and this maybe a key factor in breast cancer survival and progression. To test this hypothesis, we characterized the effect of progesterone pre-treatment on the sensitization of the epidermal growth factor (EGF) signaling pathway to EGF in the breast cancer cell line ZR-75. For the first time in ZR-75 cells and in agreement with previous work using synthetic progestins, we demonstrate that pre-treatment with the natural ligand progesterone increases EGF receptor (EGFR) levels and subsequent ligand-dependent phosphorylation. Downstream we demonstrate that progesterone alone increases erk-1 + 2 phosphorylation, potentiates EGF-phosphorylated erk-1 + 2 and maintains these levels elevated for 24 h; over 20 h longer than in vehicle treated cells. Additionally, progesterone increased the levels of STAT5, another component of the EGF signaling cascade. Progesterone increased EGF mediated transcription of a c-fos promoter reporter and the nuclear localization of the native c-fos protein. Furthermore, progesterone and EGF both alone and in combination, significantly increase cell proliferation. Several results presented herein demonstrate the conformity between the action of the natural ligand progesterone with that of synthetic progestins such as MPA and R5020 and allows the postulation that the progestin/progesterone-dependent increase of EGF signaling provides a survival advantage to burgeoning cancer cells and may contribute to the breast cancer risk associated with endogenous progesterone and with progestin-containing HRT.

Panic disorder cases in Japanese-Brazilians in Japan: their ethnic and cultural confusion.

The comparatively high salaries made in Japan are attractive to many Japanese-Brazilians. The number of individuals from this ethnic group being treated in Japanese mental hospitals has increased. We hypothesized that Japanese-Brazilian patients with panic disorders adjusted better to Japanese society and culture than those with other mental disorders. The subjects in the present study are 40 Japanese-Brazilian patients undergoing treatment at the Department of Psychiatry at Jichi Medical School, Japan, from May 1990 to September 1998. Patients were divided into a panic disorder group, a schizophrenic group, a mood disorder group and a neurosis group. Demographic data (Japanese language ability, duration of residence in Japan etc.) were collected. A comparison was made among the four groups. Patients in the panic disorder group showed a significant tendency to be fluent speakers of Japanese. Patients in the panic disorder group also had been in Japan for a significantly longer period of time than those in the other three groups. Japanese ability and length of residence in Japan rule out exacerbating factors due to a foreign living environment. Panic disorder patients usually have resolved the problems inherent in living and working in a foreign country. In general, Japanese-Brazilians are more comfortable both financially and socially in Japan than other foreign laborers because of their cultural and family background. The emotional conflict experienced by such patients may result from concern over whether to live in Brazil or Japan in the future. Their ethnic and cultural identity may be confused, fluctuating between identifying with Brazil and with Japan, and this may cause vague feelings of anxiety.

Specific detection of 16 micro-organisms in amniotic fluid by polymerase chain reaction and its correlation with preterm delivery occurrence.

OBJECTIVE: Our purpose was to develop a specific polymerase chain reaction detection method for 16 micro-organisms in amniotic fluid and to correlate its performance with bacterial cultures and preterm delivery occurrence. STUDY DESIGN: The study group was made up of 50 patients with preterm labor and intact membranes. The control group consisted of 23 patients not in labor and undergoing amniocentesis for either karyotype or lung maturity studies. Polymerase chain reaction and bacterial cultures were assayed in amniotic fluid of all patients. Results were correlated with pregnancy outcome. RESULTS: Polymerase chain reaction identified micro-organisms in 23 cases in the study group (46%), whereas cultures identified only 6 (12%). All control samples were negative for polymerase chain reaction and cultures. The sensitivity of polymerase chain reaction and cultures for the identification of patients delivering before 34 weeks' gestation was 64% and 18%, respectively. CONCLUSION: A polymerase chain reaction gene amplification method was developed to identify 16 micro-organisms in amniotic fluid. Compared with bacterial cultures, polymerase chain reaction amplification in amniotic fluid appears to be more sensitive in identifying patients delivering prematurely.

Endoscopic hemostasis of bleeding duodenal ulcer in a child with Henoch-Schönlein purpura.

A 9-year-old boy with Henoch-Schönlein purpura had a duodenal ulcer. Endoscopic injection with pure ethanol was performed on a pulsating visible vessel in the third part of the duodenum, resulting in complete hemostasis. A bleeding ulcer, although rare, may be a serious gastrointestinal complication of Henoch-Schönlein purpura and may require aggressive intervention.

Effect of omeprazole in the treatment of refractory acid-related diseases in childhood: endoscopic healing and twenty-four-hour intragastric acidity.

OBJECTIVE: To determine the clinical efficacy of once-daily treatment with omeprazole in refractory acid-related diseases in children. METHODS: Endoscopic healing and 24-hour intragastric pH values were assessed in 13 patients with refractory reflux esophagitis (n = 5), refractory and/or giant duodenal ulcer (n = 6), or giant gastric ulcer (n = 2). The mean dose of omeprazole was 0.6 mg/kg per day (range, 0.3 to 0.7 mg/kg per day). Pharmacokinetic studies of omeprazole were performed in seven patients. RESULTS: The cumulative healing rates at 2, 4, 6, and 8 weeks of treatment were 46%, 85%, 92%, and 92%, respectively. Esophagitis in one patient did not heal despite increases in doses of up to 1.6 mg/kg per day (40 mg/day). The mean intragastric pH of omeprazole-treated patients was 5.2 (range, 3.0 to 6.6) and mean hydrogenion activity was 1.78 mmol/L (range, 0.01 to 10.42 mmol/L). There was wide interindividual variation in the reduction of gastric acid production. Mean intragastric H+ activity in omeprazole-treated patients was significantly lower than that of control subjects (p < 0.005) and that of patients treated with histamine type 2(H2)-receptor antagonists (p < 0.05). Mean intragastric H+ activity was not significantly correlated to the area under the concentration-time curve of omeprazole. No severe adverse effects were reported during treatment or at follow-up. CONCLUSIONS: Omeprazole has a potent antisecretory effect and is a suitable alternative for short-term treatment of refractory acid-related diseases; a relatively low dose (0.6 mg/kg per day) appears to be optimal in most patients. Unhealed esophagitis at 8 weeks of treatment was considered to be refractory to omeprazole.

Antibiotic-associated hemorrhagic colitis without Clostridium difficile toxin in children.

We describe nine children with antibiotic-associated hemorrhagic colitis without Clostridium difficile toxin. The onset was usually sudden, with severe hematochezia and abdominal cramps. The illness quickly resolved and required no specific treatment except discontinuation of the implicated antibiotic. Early proctosigmoidoscopy was a useful diagnostic adjunct. It appears that antibiotic-associated hemorrhagic colitis is a distinct entity rather than a variant of antibiotic-associated colitis in children.

Neuron-specific enolase is a useful marker of neuroendocrine origin in pheochromocytoma cell culture

Neuron-specific enolase (NSE) has been used as a marker for neuroendocrine tumors either in immunocytochemical studies or in serum measurements. ln this paper NSE levels were determined in cultured pheochromocytoma cells to test whether it is also a useful marker ín cell culture of tumors derived from neuroendocrine system. Cultured pheochromocytoma cells came from a primary explant and were grown in RPMI supplemented with 20 per cent fetal calf serum, 100 mug/mL ampicillin and 100 mug/mL streptomycin. NSE was measured in culture medium and cell homogenates. Samples from different pheochromocytoma cultures were analyzed and compared to normal cultured fibroblast cells derived from human skin. NSE was measured by a commercially available radioimmunoassay kit. NSE levels were higher in cell homogenates as compared to those in culture medium, reaching levels as high as 6-fold in the former in TE cell line (26.46 ng/mL and 4.39 ng/mL respectively. Serial NSE measurements in culture medium from TE cell line evidenced decreasing values in subsequential subcultures (from 9.24 ng/mL during primary explant to 1.7 ng/ml. in the 10th subculture). In cultured normal fibroblasts, NSE levels in cultured media were definitely lower than those obtained from pheochromocytoma cultures. These preliminary data suggest that NSE may be a useful marker of neuroendocrine derived tumors, such as pheochromocytoma, in culture. Thus, the simplicity and availability of NSE radioimmunoassay provides an alternative to catecholamine measurement to better characterize pheochromocytoma cell lines in culture, with the advantage of faster results at lower costs.

Fatores que influenciam o sucesso em cultura de fibroblastos. / Factors influencing the sucess in fibroblasts culture

Relataram-se diferentes fatores que podem influenciar no sucesso do estabelecimento do metodo de cultura de fibroblastos. Tres linhagens celulares foram cultivadas advindas de: pele de embriao de galinha, fibroblastos de rato (ST1) e pele humana normal. Mostrou-se a evolucao inicial do crescimento celular de explantes primarios de pele humana e de embriao de galinha