Gonadotropin-releasing hormone antagonists increase follicular fluid insulin-like growth factor-I and vascular endothelial growth factor during ovarian stimulation cycles.

The aim of the present study was to investigate the effect of gonadotropin-releasing hormone (GnRH) antagonists (GnRH-ant) on follicular fluid (FF) insulin-like growth factor-I (IGF-I) and FF vascular endothelial growth factor (VEGF) levels. Sixty women undergoing assisted reproduction were randomized and assigned to two different GnRH analog regimens: GnRH agonist (GnRH-a) and GnRH-ant. FF VEGF and FF IGF-I concentrations were significantly increased in the patients treated with GnRH-ant (p < 0.001). In the same patients we observed a statistically significant reduction in serum luteinizing hormone (LH) and estradiol (E2) levels (p < 0.001 and p < 0.05, respectively), FF E2 and FF androstenedione levels (p < 0.05 and p < 0.001, respectively), as well as a reduction in the number of pregnancies although this was not statistically significant. In the GnRH-ant group, FF VEGF levels were positively correlated with FF IGF-I levels, and both were negatively correlated with serum LH levels. The increase in FF IGF-I and FF VEGF levels in women treated with GnRH-ant could be explained by a deleterious follicular environment in response to profound suppression of LH and E2 levels.     

Proximal osteotomy of the tibia for the treatment of genu recurvatum in adults.

Twenty-seven opening-wedge osteotomies of the proximal part of the tibia were performed in twenty-five patients who had genu recurvatum. In sixteen knees, the genu recurvatum was due entirely to osseous deformity. In the remaining eleven knees, it was due to a combination of osseous and soft-tissue deformity; in five, the deformity was predominantly osseous and in six, primarily in the soft tissues (the ligaments and capsule). The average age of the patients was twenty-three years (range, fifteen to fifty-four years). The osteotomy was proximal to the tibial tuberosity in twenty-two knees. In eighteen of these knees, the tuberosity was detached with its patellar ligament and then reattached to the proximal part of the tibia over the block of bone in the opened wedge; in the remaining four knees, the tibial tuberosity was not detached. The osteotomy was distal to the tuberosity in five knees. The patients were followed for an average of 14.5 years (range, three to thirty years). Of the eighteen knees in which the osteotomy had been proximal to the tibial tuberosity and the tuberosity had been detached and then reattached, nine (50 per cent) had a result that was excellent; five (28 per cent), good; and four (22 per cent), fair. Of the four knees in which the operation had been proximal to but without detachment of the tuberosity, one had a result that was excellent; two, good; and one, fair. Of the five knees in which the osteotomy was distal to the tibial tuberosity, one had a result that was good; three, fair; and one, poor. Of the twenty-one knees in which the deformity was entirely or predominantly osseous, eighteen (86 per cent) had an excellent or good result. None of the six knees in which the deformity was predominantly in the soft tissues had an excellent or good result. Patients in whom the deformity was not primarily osseous, and those in whom the operation was distal to the tibial tubercle, were much more likely to have a fair or poor result.     

Cytotoxic activity of cardenolides from Beaumontia brevituba stems.

Five known cardenolides, digitoxigenin (1), oleandrigenin (2), digitoxigenin alpha-L-cymaroside (3), digitoxigenin beta-gentiobiosyl-alpha-L-cymaroside (4), and delta 16-digitoxigenin beta-D-glucosyl-alpha-L-cymaroside (5), were isolated from the stems of Beaumontia brevituba Oliver by cytotoxicity-directed fractionation monitored by a cultured human lung cancer cell line. The cytotoxic activity of these compounds was evaluated with a panel of twelve human and murine cancer cell lines. The lignan glycoside, syringaresinol beta-D-glucoside, was obtained for the first time in the form of its levo-enantiomer.     

Assessment of cyclooxygenase inhibitors using in vitro assay systems

Various inhibitors of cyclooxygenase are known to mediate cancer chemopreventive effects. We currently describe two in vitro assay systems for measuring cyclooxygenase activity. These assays can be used in combination and thereby provide a rapid, reliable, and economical approach that is applicable for large-scale evaluation of test samples. This approach employs peroxidase co-substrate oxidation and oxygen consumption assays. The former system, adapted to a 96-well plate format, detects inhibitors that function as a radical scavengers or interact with the enzyme directly. The latter system specifically monitors cyclooxygenase inhibitors that interact with the enzyme itself. Thus, the peroxidase co-substrate oxidation assay serves as a pre-screening method, whereas the oxygen consumption assay is used subsequently to investigate the mode of action mediated by samples which test positive.     

Enhancement of benzo(a)pyrene metabolism mediated by retinol acetate in cultured human bronchial epithelial-cells

As suggested by animal studies and human epidemiological data, retinoids possess significant cancer chemopreventive activity. Although the majority of studies in this area have focused on the ability of retinoids to prevent the promotion or progression of carcinogenesis, a significant amount of data suggest retinoids can alter initiation events. In the current report, we have evaluated the potential of retinol acetate to modulate benzo(a)pyrene metabolism in low-passage human bronchial epithelial cells in monolayer cultures, Of 16 different cell cultures, benzo(a)pyrene metabolism was increased in 14, decreased in one, and unchanged in one, when retinol acetate was added to the media, In a preliminary study with one of the cell cultures in which retinol acetate significantly enhanced benzo(a)pyrene metabolism, binding of carcinogen metabolites to;DNA was unaffected, Since retinoids are known cancer chemopreventive agents and carcinogen binding to DNA is the key event in the initiation of carcinogenesis, these results suggest that retinoids may decrease carcinogenic risk by increasing the detoxification of procarcinogens such as benzo(a)pyrene ina manner that does not yield a concomitant increase in damage to critical cellular targets such as DNA.     

Multiple myeloma regression mediated by bruceantin.

PURPOSE: Bruceantin has been shown to induce cell differentiation in a number of leukemia and lymphoma cell lines. It also down-regulated c-MYC, suggesting a correlation of down-regulation with induction of cell differentiation or cell death. In the present study, we focused on multiple myeloma, using the RPMI 8226 cell line as a model. EXPERIMENTAL DESIGN: The effects of bruceantin on c-MYC levels and apoptosis were examined by immunoblotting, 4',6-diamidino-2-phenylindole staining, evaluation of caspase-like activity, and 3,3'-dihexyloxacarbocyanine iodide staining. The potential of bruceantin to inhibit primary tumor growth was assessed with RPMI 8226 xenografts in SCID mice, and apoptosis in the tumors was evaluated by the terminal deoxynucleotidyl transferase-mediated nick end labeling assay. RESULTS: c-MYC was strongly down-regulated in cultured RPMI 8226 cells by treatment with bruceantin for 24 h. With U266 and H929 cells, bruceantin did not regulate c-MYC in this manner. Apoptosis was induced in the three cell lines. In RPMI 8226 cells, apoptosis occurred through proteolytic processing of procaspases and degradation of poly(ADP-ribose) polymerase. The mitochondrial pathway was also involved. Because RPMI 8226 cells were the most sensitive, they were used in a xenograft model. Bruceantin treatment (2.5-5 mg/kg) resulted in a significant regression of tumors without overt toxicity. Apoptosis was significantly elevated in tumors derived from animals treated with bruceantin (37%) as compared with the control tumors (14%). CONCLUSIONS: Bruceantin interferes with the growth of RPMI 8226 cells in cell culture and xenograft models. These results suggest that bruceantin should be reinvestigated for clinical efficacy against multiple myeloma and other hematological malignancies.     

Betulinic acid-induced programmed cell death in human melanoma cells involves mitogen-activated protein kinase activation.

Betulinic acid, a naturally occurring triterpene found in the bark of the white birch tree, has been demonstrated to induce programmed cell death with melanoma and certain neuroectodermal tumor cells. We demonstrate currently that treatment of cultured UISO-Mel-1 (human melanoma cells) with betulinic acid leads to the activation of p38 and stress activated protein kinase/c-Jun NH(2)-terminal kinase [widely accepted proapoptotic mitogen-activated protein kinases (MAPKs)] with no change in the phosphorylation of extracellular signal-regulated kinases (antiapoptotic MAPK). Moreover, these results support a link between the MAPKs and reactive oxygen species (ROS). As demonstrated previously, cells treated with betulinic acid generate ROS. Preincubation of cells with antioxidants blocks the process of programmed cell death, and prevents the phosphorylation of p38 and stress activated protein kinase/c-Jun NH(2)-terminal kinase. These data suggest that ROS act upstream of the MAPKs in the signaling pathway of betulinic acid. In addition to mediating these responses, treatment of cells with betulinic acid resulted in a gradual depolarization of mitochondrial membrane potential, a phenomenon established to contribute to the induction of programmed cell death. Interestingly, p38 was capable of partially modulating this perturbation, and investigations of mitochondria-associated apoptotic events indicate no involvement of known caspases. These data provide additional insight in regard to the mechanism by which betulinic acid induces programmed cell death in cultured human melanoma cells, and it likely that similar responses contribute to the antitumor effect mediated with human melanoma carried in athymic mice.     

Resveratrol inhibits rhabdomyosarcoma cell proliferation.

Rhabdomysarcoma is the most common soft tissue tumour in children under the age of 15. Although the introduction of multimodal treatment programmes, including chemotherapy, radiation therapy and excision have increased the overall survival, the chemotherapeutic agents currently used for the treatment of rhabdomyosarcoma exhibit considerable toxicity. The aim of this study was to investigate the effects and possible mechanism(s) of action of resveratrol on human embryonal rhabdomyosarcoma (RD) cells. Resveratrol is a natural polyphenolic compound produced in a number of edible plants and has received considerable attention as a potential chemopreventive and/or chemotherapeutic agent against various types of cancers. In the present study, resveratrol was shown to inhibit cell proliferation of RD cells in a dose-dependent manner with an IC50 of 48.1 micromol/l and induce an arrest in the S/G2 phase of the cell cycle. As evident from immunocytochemical data, resveratrol treatment increased the size of the RD cells. Furthermore, resveratrol treatment resulted in a significant downregulation of cyclin B expression as demonstrated by western blot analyses. In conclusion, the present study shows that resveratrol exerts a strong inhibition of rhabdomyosarcoma cell proliferation in part by arresting cells in S/G2 phase of the cell cycle. These findings warrant further investigation to establish potential use of resveratrol as a relatively non-toxic chemotherapeutic agent for the treatment of rhabdomyosarcoma.     

Dietary Administration of Asimina triloba. (Paw Paw) Extract Increases Tumor Latency in N.-Methyl-N.-nitrosourea–Treated Rats

Abstract

The paw paw tree, Asimina triloba. (L.) Dunal (Annonaceae), contains more than 50 bioactive components, primarily annonaceous acetogenins. Some therapeutic activities have been associated with this material, but the potential to mediate a cancer chemopreventive effect has not been reported. In this study, a standardized extract from the twigs, in which bullatacin, asimicin, and trilobacin represent the most potent and major bioactive acetogenins, was tested in the N.-methyl-N.-nitrosourea–induced mammary carcinogenesis model. With Sprague-Dawley rats given a diet containing paw paw extract (1250 and 2500 mg/kg diet; based on maximum tolerated dose studies), mammary tumor latency was increased from 55 to 66 days. However, mammary tumor incidence and multiplicity were not affected by extract consumption.