Green Tropical Phytoextracts - Promising Anticancer Alternative

ABSTRACT The anticancer potential of aqueous extracts of tropical plants Ficus deltoidea and Labisia pumila were tested on human prostate carcinoma (DU145) cells. The effects of these extracts on the cells were observed through several tests such as cytotoxicity MTT assay, trypan blue exclusion assay (for cell viability), fluorescence and morphological observations. Both of the plant extracts had a growth repression effect on the cells with different concentrations. The best concentration (1x10-3 μg/mL) was chosen by determining their cytotoxic effects on human prostate cancer as well as fibroblast (normal) cells. The F. deltoidea extract showed more potency and killed the cells faster than L. pumila. Morphologically both extracts affected the cells in a similar way, i.e. the cells became irregular shaped, detached themselves and kept floating in the medium.

Evaluation of a haemostatic agent in rabbits.

Topical hemostatic agents are applied locally to areas of injured vascular endothelium to control local bleeding. Ankaferd Blood Stopper (ABS) has gained approval in Turkey and Bosnia-Herzegovina as a topical haemostatic agent for external post-surgical and post-dental surgery bleeding. The safety of topical use of ABS has been demonstrated in numerous in vitro and in vivo animal models, as well as in a clinical Phase I trial in humans. ABS, besides its haemostatic activity, also has in vitro anti-infectious and anti-neoplastic effects. To assess potential detrimental effects of intravenous administration of ABS into intact systemic circulation in a rabbit experimental model, one milliliter of ABS was administered intravenously into the systemic circulation of twelve rabbits which were included in the study via the marginal ear vein. Animals were observed for 1 hr before euthanasia was performed by administering 40 mg of intracardiac suxamethonium chloride. In the event of death (cardiopulmonary arrest) before the end of the planned observation period of 60 minutes, time of death was recorded and histopathological examination of the liver and spleen was commenced. Ten rabbits were alive by the end of the planned observation period, without showing any clear signs of discomfort, whereas two animals died within five minutes after systemic administration of intravenous ABS. Postmortem histopathological examination of the livers and spleens of all animals' revealed findings consistent with hepatic venous outflow obstruction. Systemic intravascular administration of ABS into intact vascular endothelium should never be performed in any setting. Further experimental and clinical studies on this liquid hemostatic agent should proceed by accepting ABS as purely a topical haemostatic agent, to be applied solely to areas of injured vascular endothelium.

Oral systemic administration of Ankaferd blood stopper has no short-term toxicity in an in vivo rabbit experimental model.

BACKGROUND: Ankaferd blood stopper (ABS) is a standardized herbal extract obtained from 5 different plants. In Turkey, it has been approved for local topical applications in external postsurgical and postdental surgery bleedings. Ankaferd blood stopper, besides its hemostatic activity, has in vitro anti-infectious and antineoplastic actions. OBJECTIVE: The aim of this study was to assess short-term hematological and biochemical safety following the oral systemic administration of ABS to rabbits. METHODS: Twelve rabbits (aged 6-12 months) were included to test the safety of oral ABS. Animals were divided into 4 groups, which had ABS administered orally at doses of 1, 3, 6, and 9 mL, irrespective of their weight. The general well-being and feeding patterns of the animals were observed for a period of 7 days. Blood samples (5.5 mL) were obtained just before oral administration, on days 1 and 4. RESULTS: During the observation period of 7 days, none of the animals showed any abnormal behavior or deviation from the normal. Acute mucosal toxicity, hematotoxicity, hepatotoxicity, nephrotoxicity, and biochemical toxicity were not observed during the short-term follow-up of the animals. CONCLUSIONS: No signs of toxicity were observed in rabbits during short-term study with oral ABS administration.

Ultrastructural and morphological analyses of the in vitro and in vivo hemostatic effects of Ankaferd Blood Stopper.

Ultrastructural and morphological analyses of a novel hemostatic agent, Ankaferd Blood Stopper (ABS), in comparison to its in vitro and in vivo hemostatic effects were investigated. High-resolution scanning electron microscopy (SEM) images accompanied with morphological analysis after topical application of ABS revealed a very rapid (<1 second) protein network formation within concurrent vital erythroid aggregation covering the classical coagulation cascade. Histopathological examination revealed similar in vivo ABS-induced hemostatic network at the porcine hepatic tissue injury model. Instantaneous control of bleeding was achieved in human surgery-induced dental tissue injury associated with primary and secondary hemostatic abnormalities. Ankaferd Blood Stopper could hold a great premise for clinical management of surgery bleedings as well as immediate cessation of bleeding on external injuries based on upcoming clinical trials.

Effects of a topical hemostatic agent on an epistaxis model in rabbits.

BACKGROUND: Ankaferd BloodStopper (ABS) is a standardized herbal compound consisting of 5 different plant extracts. ABS, as a topical medicinal product, has been approved by the Turkish Ministry of Health for the management of dermal, external postsurgical and postdental surgery bleedings. OBJECTIVE: The aim of this study was to assess the hemostatic effect of ABS in an experimental epistaxis model. METHODS: Adult female New Zealand rabbits were used in this study. Standardized full-thickness mucosal wounds were created to provide bleeding on the right and left side of the nasal septum of rabbits with a 3-mm surgical punch. To stop bleeding, the wounds were treated with topical ABS on one side and with topical isotonic saline solution as a control on the other side of the nasal septum. Investigators were blinded to treatment. The duration of bleeding was measured in order to compare the hemo-static effect with ABS or topical isotonic saline solution. The wounds were observed for the duration of bleeding in order to compare the hemostatic effect of ABS with isotonic saline solution and acute adverse effects (AEs) for 30 minutes by a researcher blinded to treatment groups. RESULTS: Six rabbits (mean weight, 2.6 kg [range, 2.3-3.1 kg]) were used in this study. Mean (SD) bleeding time in the wounds administered ABS (98 [17] sec) was significantly less than that of the controls (266 [36] sec; P = 0.004). No AEs were observed up to 30 minutes after study initiation. CONCLUSION: This study suggests that ABS may be more effective in reducing time to hemostasis when compared with isotonic saline solution in this epistaxis model in rabbits.

The efficacy of Ankaferd Blood Stopper in antithrombotic drug-induced primary and secondary hemostatic abnormalities of a rat-bleeding model.

Ankaferd comprises a standardized mixture of plants Thymus vulgaris, Glycyrrhiza glabra, Vitis vinifera, Alpinia officinarum and Urtica dioica. Ankaferd Blood Stopper (ABS) as a medicinal product has been approved in the management of external hemorrhage and dental surgery bleedings in Turkey. This study aimed to evaluate the in-vivo hemostatic effect of ABS in rats pretreated with acetylsalicylic acid or enoxaparin. Wistar rats (210-270 g) of both sexes were used in this study. The animals were pretreated with acetylsalicylic acid (10 mg/kg) orally for 4 days or enoxaparin sodium (8 mg/kg) subcutaneously for 3 days or did not receive any anticoagulant before tail cut at 4th day. ABS was administered topically [a total of 4 ml (1 ml/puff x 4)] to the cut tail in the studied animals. The duration of bleeding and the amount of bleeding were measured in order to evaluate the hemostatic effect of ABS. In acetylsalicylic acid-treated animals, topical ABS reduced both the duration and also the amount of bleeding volume by 68.4 and 54.6%, respectively. It was also effective in shortening the duration of bleeding (30.6%) and decreasing the amount of bleeding (32.8%) in enoxaparin-treated animals. ABS, a traditional folkloric medicinal plant extract, has in-vivo hemostatic actions, which may provide a therapeutic potential for the management of patients with deficient hemostasis in the clinical medicine.

Hemostatic efficacy of Ankaferd Blood Stopper in a swine bleeding model.

OBJECTIVE: The purpose of this study was to show the hemostatic effect of spray, solution and tampon forms of Ankaferd Blood Stopper (ABS), a unique medicinal plant extract historically used as a hemostatic agent in Turkish folklore medicine, in a porcine bleeding model. MATERIALS AND METHODS: Two 1-year-old pigs were used as bleeding models for superficial and deep skin lacerations, grade II liver and spleen injuries, grade II saphenous vein injury and grade IV saphenous artery injury. Spray, solution or tampon forms of ABS were applied after continuing bleeding was confirmed. The primary outcome was time to hemostasis. Volume of blood loss was not measured. The pigs were euthanized at the end of the experiment. RESULTS: Spray or direct application of ABS solution resulted in instant control of bleeding in superficial and deep skin lacerations as well as puncture wounds of the liver. A 40-second application of ABS tampon was sufficient to stop bleeding of skin lacerations, while 1.5- and 3.5-min applications were used to control hemorrhage from the saphenous vein and artery, respectively. No rebleeding was observed once hemostasis was achieved. However, repeated applications of ABS solution and tampon were only temporarily effective in the hemostasis of spleen injury. CONCLUSIONS: The data showed that ABS was an effective hemostatic agent for superficial and deep skin lacerations and minor/moderate trauma injuries in a porcine bleeding model.

In vivo hemostatic effect of the medicinal plant extract Ankaferd Blood Stopper in rats pretreated with warfarin.

AIM: Ankaferd comprises a mixture of Thymus vulgaris, Glycyrrhiza glabra, Vitis vinifera, Alpinia officinarum and Urtica dioica. Ankaferd Blood Stopper (ABS) has been approved in the management of bleedings. This study aimed to evaluate in vivo hemostatic effect of ABS in rats pretreated with warfarin. MATERIALS AND METHODS: Wistar rats (210-270 g) were treated either with warfarin (2 mg/kg) or vehicle (0.9% NaCl) orally before bilateral hind leg amputation. ABS was administered topically to one of the amputed legs. The duration of bleeding and the amount of bleeding were measured to evaluate the hemostatic effect of ABS. RESULTS: Topical ABS administration to amputed leg shortened the duration of bleeding markedly in both untreated and warfarin-treated rats by 31.9% [1.42 min (95% CI: 0.35-2.49)] and 43.5% [5.12 min (95% CI: 2.16-8.07)] respectively. The amount of bleeding in ABS-administered amputed leg showed a decrease by 53.8% in warfarin-treated group. CONCLUSIONS: ABS has in vivo hemostatic actions that may provide a therapeutic potential for the management of patients with deficient primary hemostasis in clinical medicine.

Superoxide dismutase and glutathione peroxidase in erythrocytes of patients with iron deficiency anemia: effects of different treatment modalities.

AIM: To test whether the activities of erythrocyte superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) can be affected by oral iron (OI) treatment, parenteral iron (PI) treatment, and parenteral iron treatment with vitamin E supplementation (PIE) in iron deficiency anemia. METHODS: Twenty-eight patients with iron deficiency anemia and 18 healthy controls were included in the study. Anemic patients were systematically allocated into 3 treatment groups. The first group (n=8) received OI, the second group (n=10) PI, and the third group (n=10) PIE. SOD and GSH-Px activities were determined using commercial kits. RESULTS: Before the treatment, SOD activity was significantly lower in anemic patients than in the control group (Kruskal-Wallis test, p<0.05). After the treatment, SOD activity significantly increased in all three patient groups and reached the values found in the control group (Wicoxon signed-rank test, p=0.017 for OI, p=0.047 for PI, and p=0.037 for PIE group). Before the treatment, GSH-Px activities in anemic patients were similar to that of control group (Kruskal-Wallis test, p>0.05). Although there was no significant decrease in GSH-Px activity after OI treatment, both PI and PIE treatments significantly decreased GSH-Px activity (Wilcoxon signed-ranks test, p=0.007 for PI and p=0.005 for PIE). PIE was more effective than PI treatment in maintaining GSH-PX activity. CONCLUSION: Oral iron treatment improved the iron deficiency anemia and recovered antioxidant defense system by increasing SOD activity and maintaining GSH-Px activity at normal level. When parenteral iron treatment is inevitable, vitamin E supplementation together with PI treatment may be effective in partially restoring the antioxidant status in this type of anemia.