Immunolocalization of VEGF, VEGF receptors, EGF-R and Ki-67 in leiomyoma, cellular leiomyoma and leiomyosarcoma.

Angiogenic factors, such as vascular endothelial growth factor (VEGF), its receptors and epidermal growth factor receptor (EGF-R), are involved in increased progression in many carcinomas. The aim of this study was to investigate the role of angiogenesis and immunolocalization of VEGF, its receptors, EGF-R and Ki 67 in leiomyomas and leiomyosarcomas using an indirect immunohistochemical method. Samples from patients with leiomyoma, cellular leiomyoma and cellular leiomyosarcoma (n=20 per group) were fixed in 10% formalin and processed using routine paraffin protocols. Following initial histological analysis, samples were immunostained with primary antibodies for VEGF, VEGFR-1, VEGFR-2, EGF-R and Ki-67 using an indirect avidin-biotin peroxidase method. Immunostaining intensities were evaluated as mild, moderate or strong and a semi-quantitative method (H-Score) was used to compare the samples. While mild/moderate EGF-R immunostaining and moderate immunostaining for VEGF and its receptors were observed in samples of leiomyomas, much less immunoreactivity was observed in cellular leiomyomas. All immunoreactivities and immune-stained cells increased in leiomyosarcomas. When scores of intensity and percentage of positive staining cells were compared, all immunoreactivities were shown to be significantly increased in leiomyosarcomas compared to leiomyomas. These results suggest that in leiomyosarcoma, angiogenic factors, such as VEGF, its receptors and EGF-R, may be involved in tumor angiogenesis. Active tumor cells can trigger angiogenesis, interaction with surrounding tissue and in the tissue itself initiating angiogenic activity. Angiogenic growth factors play an important role and induce malignant transformation through both autocrine and paracrine mechanisms. Anti-angiogenic agents may provide a novel therapeutic approach for the treatment of leiomyosarcoma.

Levosimendan up-regulates transforming growth factor-beta and smad signaling in the aorta in the early stage of sepsis.

BACKGROUND: This prospective, controlled experimental study was planned to investigate the effects of levosimendan on transforming growth factor (TGF)-beta3 and Smad1, Smad2 and Smad3 expression in the early stages of sepsis. METHODS: Twenty-four rats were randomized into four groups: (1) sham-operated controls, (2) dobutamine group--subjected to abdominal hypertension and peritonitis-induced sepsis using cecal ligation and puncture (CLP), then treated with 10 microg x kg(-1) min(-1) intravenous (IV) dobutamine infusion, (3) levosimendan group--as in 2, then treated with levosimendan IV bolus 200 microg x kg(-1) followed by 200 microg x kq(-1) min(-1) IV infusion, and (4) a control group as in 2, with no treatment. All rats were killed 8 hours after CLP. Aorta tissue samples were analyzed by immunohistochemical staining. RESULTS: CLP caused mild interleukin (IL)-1 immunostaining in both control and dobutamine groups. Immunoreactivity of tumor necrosis factor (TNF)-alpha was mild in both sham and control groups. TGF-beta3 immunostaining was mildly increased in groups sham, control and dobutamine, whereas it was found moderate in group levosimendan. Smad1, Smad2 and Smad3 were found moderately increased only in group levosimendan. CONCLUSION: Beneficial effects of levosimendan on hemodynamics and global oxygen transport were reported in experimental and clinical trials. Besides its potency on C++ ion sensitivity, it should influence inflammatory cytokine production by diminishing TGF-beta3 and Smad1, Smad2 and Smad3 expression.

Antibiotic treatment is superior to ursodeoxycholic acid on total parenteral nutrition associated hepatic dysfunction.

PURPOSE: This study aimed to investigate the apoptotic mechanisms, oxidative stress, and mechanisms of effect of antibiotics and ursodeoxycholic acid (UDCA) in total parenteral nutrition (TPN)-associated liver injury. METHODS: Four groups of young rabbits were used in the study as follows: Group 1 (n: 7): TPN + Metronidazole (30 mg/kg IV) + Gentamicin (6 mg/kg IV); Group 2 (n: 7): TPN + UDCA (15 mg/kg per oral); Group 3 (n: 6): TPN only; and Group 4 (n: 7): Control group. After 10 days, the animals were killed and livers were removed. Hepatic apoptosis, apoptotic proteins, malondialdehyde (MDA) and myeloperoxidase (MPO) levels were studied in liver, and direct bilirubin values were assessed in the blood samples. RESULTS: Direct bilirubin increased with TPN, and antibiotic combination, as the most effective group, significantly lowered its levels (p < 0.01). MDA values also showed significant differences in comparisons between G1 and G3 (p < 0.05) and G1-G4 (p < 0.01). An increased number of apoptotic cells was detected particularly in G2 and G3, whereas the lowest levels, other than in the control group, were found in G1. All TUNEL-positive cell number data were statistically significant except between G2 and G3(p < 0.05). Caspase-3 and Bax immunoreactivities were greatest in G2. Significant differences were shown in caspase-3 immunoreactivity between the groups (p < 0.01), except between G1 and G3 (p > 0.05). All comparisons between the groups were significant for Bax (p < 0.01). In contrast, Bcl-2 immunoreactivity was moderate and highest in G1: comparisons between G1 and the other groups demonstrated statistically significant differences (p < 0.01). Fas-L immunoreactivity was greatest in G2, and all comparisons between the groups were statistically significant (p < 0.01). CONCLUSIONS: Metronidazole and gentamicin combination is effective on TPN-induced liver injury by the Bcl-2 anti-apoptotic pathway, total anti-apoptotic effect and by decreasing bilirubin levels. Oxidative injury in the liver increased with therapy. UDCA seems less effective on TPN-associated liver injury.

Propolis from Turkey induces apoptosis through activating caspases in human breast carcinoma cell lines.

Propolis is a sticky substance that is collected from plants by honeybees that has anti-mutagenic and anti-carcinogenic properties with biological and therapeutic effects. The target of this study was to investigate the anti-apoptotic effect of propolis extracts (PE) on the caspase pathway in the human breast cell line MCF-7 in culture. Seven different propolis extracts, numbered PE 1-7, produced in their natural ecological environment, were collected from the Hacettepe University Beytepe Campus area in Ankara, Turkey. Individual extracts at 0.5, 0.25, 0.125 and 0.063mg/ml were incubated with MCF-7 cells during 2 days culture. Cell growth and cytotoxicity were measured colorimetrically by MTT assay. Apoptotic cell death was determined by the TUNEL method (terminal deoxynucleotidyltransferase-biotin nick end-labelling) and caspase activity was investigated by immunocytochemistry using antibodies directed against caspase 6, caspase 8 and caspase 9. The results showed that the PE 5 and 6 extracts at 0.125mg/ml dilution induced apoptosis in association with increased number of TUNEL positive cells. MTT results showed that cultures exposed to the same extracts and at the same dilution experienced better cell growth compared to those cultures exposed to the other extracts. Immunpositivity for all caspases was detected after treatment with all the extracts and at all dilutions, with stronger immunoreactivity for caspase 6 than caspases 8 and 9. Caspase 6 labelling was especially strong in PE 5 and PE 6. We conclude that propolis may have anti-tumour effects by increasing apoptosis through the caspase pathway. Such propolis extracts may be important economically and allow development of a relatively inexpensive cancer treatment.