Phytochemical Analysis and Anticancer Properties of Drimia maritima Bulb Extracts on Colorectal Cancer Cells.

Cancer is a worldwide health problem and is the second leading cause of death after heart disease. Due to the high cost and severe side effects associated with chemotherapy treatments, natural products with anticancer therapeutic potential may play a promising role in anticancer therapy. The purpose of this study was to investigate the cytotoxic and apoptotic characteristics of the aqueous Drimia maritima bulb extract on Caco-2 and COLO-205 colorectal cancer cells. In order to reach such a purpose, the chemical composition was examined using the GC-MS method, and the selective antiproliferative effect was determined in colon cancer cell lines in normal gingival fibroblasts. The intracellular ROS, mitochondrial membrane potential, and gene expression changes in selected genes (CASP8, TNF-α, and IL-6 genes) were assessed to determine the molecular mechanism of the antitumor effect of the extract. GC-MS results revealed the presence of fifty-seven compounds, and Proscillaridin A was the predominant secondary metabolite in the extract. The IC50 of D. maritima bulb extract on Caco-2, COLO-205, and the normal human gingival fibroblasts were obtained at 0.9 µg/mL, 2.3 µg/mL, and 13.1 µg/mL, respectively. The apoptotic effect assay indicated that the bulb extract induced apoptosis in both colon cancer cell lines. D. maritima bulb extract was only able to induce statistically significant ROS levels in COLO-205 cells in a dose-dependent manner. The mitochondrial membrane potential (MMP) revealed a significant decrease in the MMP of Caco-2 and COLO-205 to various concentrations of the bulb extract. At the molecular level, RT-qPCR was used to assess gene expression of CASP8, TNF-α, and IL-6 genes in Caco-2 and COLO-205 cancer cells. The results showed that the expression of pro-inflammatory genes TNF-α and IL-6 were upregulated. The apoptotic initiator gene CASP8 was also upregulated in the Caco-2 cell line and did not reach significance in COLO-205 cells. These results lead to the conclusion that D. maritima extract induced cell death in both cell lines and may have the potential to be used in CRC therapy in the future.

Green Synthesis of Gold, Iron and Selenium Nanoparticles Using Phytoconstituents: Preliminary Evaluation of Antioxidant and Biocompatibility Potential.

This study aimed at fabricating gold (Au), iron (Fe) and selenium (Se) nanoparticles (NPs) using various natural plant extracts from the Fertile Crescent area and evaluating their potential application as antioxidant and biocompatible agents to be used in the pharmaceutical field, especially in drug delivery. The Au-NPs were synthesized using Ephedra alata and Pistacia lentiscus extracts, whereas the Fe-NPs and Se-NPs were synthesized using peel, fruit and seed extracts of Punica granatum. The phytofabricated NPs were characterized by the UV-visible spectroscopy, scanning electron microscope, Fourier transform infrared spectroscopy, X-ray diffraction (XRD) and energy-dispersive X-ray (EDS) spectroscopy. Scanning electron microscope technique showed that the synthesized NPs surface was spherical, and the particle size analysis confirmed a particle size of 50 nm. The crystalline nature of the NPs was confirmed by the XRD analysis. All synthesized NPs were found to be biocompatible in the fibroblast and human erythroleukemic cell lines. Se-NPs showed a dose-dependent antitumor activity as evidenced from the experimental results with breast cancer (MCF-7) cells. A dose-dependent, free-radical scavenging effect of the Au-NPs and Se-NPs was observed in the DPPH (2,2-Diphenyl-1-picrylhydrazyl) assay, with the highest effect recorded for Au-NPs.

Ononis natrix L. Lowers the Blood Glucose Concentration in Wistar Rats with Streptozotocin-induced Diabetes Mellitus.

BACKGROUND: Practitioners of traditional medicine use the decoction of Ononis natrix L. to treat hyperglycemia. The literature offers no evidence to support the use. OBJECTIVE: To investigate the effect of the decoction of Ononis natrix L. on the blood glucose concentration in Wistar rats (Rattus norvegicus) with streptozotocin-induced diabetes mellitus. METHODS: We obtained 35 Wistar rats from the animal colony of The University of Jordan School of Medicine. We induced diabetes by a single intraperitoneal injection of streptozotocin (60 mg/kg body weight) and 23 rats (66%) survived to allocation. We randomly assigned the rats to one of four groups: negative control (1% Tween 80 in distilled water), positive control (100 mg/kg metformin), high-dose treatment (7.5 mL of the decoction), and low-dose treatment (3.5 mL of the decoction). We administered the doses twice daily by oral gavage for two weeks and measured the tailblood glucose concentration twice daily, once before the first dose and another time after the second dose. We used linear mixed-effects regression to model the change in blood glucose concentration as a function of the experimentation groups, with adjustments for pseudoreplication and temporal variation. RESULTS: The estimated mean change was 1 mmol/L (-30 to 31 mmol/L) for the negative control group, -26 mmol/L (-56 to 5 mmol/L) for the positive control group, -75 mmol/L (-108 to -42) for the low-dose treatment group, and -82 mmol/L (-111 to -53 mmol/L) for the high-dose treatment group. CONCLUSION: In conclusion, we demonstrate, for the first time, the hypoglycemic effect of Ononis natrix L. in an animal model of diabetes.

Vitamin B12 Deficiency Among the Healthy Jordanian Adult Population: Diagnostic Levels, Symptomology and Risk Factors.

BACKGROUND: Compared to the data available for developed countries, there is a marked scarcity of information on the levels and symptomology of vitamin B12 deficiency in developing countries, particularly in the Middle Eastern region. OBJECTIVE: To explore (a) the risk factors associated with a deficiency of vitamin B12, and (b) the baseline (cut-off) serum level of vitamin B12 for a clinically-symptomatic deficiency in the Jordanian adult population. METHODS: A total of 485 subjects were included in this study. Blood samples were drawn for biochemical analysis and data regarding socio-demographics, general health, anthropometric measures, and past medical, surgical, and medication history were collected. To explore the cut-off point, we compared all parameters included in a standard complete blood count as well as the main symptoms reported to be associated with B12 deficiency between groups of different B12 cut-off values, consisting of those above and below 200, 175, 150 and 125 pg/ml. RESULTS: Low dairy intake habits, age, recurrent headaches, heartburn, and peptic ulcer disease were found to be significantly associated with lower vitamin B12 levels. Surprisingly, daily smoking was associated with significantly higher B12 levels. The results revealed that none of the included potential indicators of B12 deficiency could be considered an indicative feature of deficiency. There were no significant differences neither in the symptoms nor in the CBC parameters between any of the tested study groups. CONCLUSION: Low dairy intake, older ages, recurrent headaches, heartburn, and peptic ulcer disease all could be considered as risk factors of having low vitamin B12 levels within the Jordanians. Also, they tend to have lower levels of vitamin B12 levels, in comparison to countries in the West, without necessarily having deficiency symptoms. The cut-off value to diagnose functional B12 deficiency could be less than 125 pg/ml for the Jordanians. More local studies are needed to establish an accurate vitamin B12 cut-off value for the population in Jordan.

Ethanol Extract of Achillea fragrantissima Enhances Angiogenesis through Stimulation of VEGF Production.

OBJECTIVE: Achillea fragrantissima L. (Asteraceae) is a traditionally used medicinal herb in the rural communities of Jordan. METHODS: The present study evaluated the efficacy of the ethanol extract of this species on angiogenesis in both, ex vivo using a rat aortic ring assay and in vivo using a rat excision wound model. RESULTS: In concentrations of 50 and 100 µg/ml, the ethanol extract showed angiogenic stimulatory effect and significantly increased length of capillary protrusions around aorta rings of about 60% in comparison to those of untreated aorta rings. In MCF-7 cells, the ethanol extract of A. fragrantissima stimulated the production of VEGF in a dose-dependent manner. 1% and 5% of ethanol extract of A. fragrantissima containing vaseline based ointment was applied on rat excision wounds for six days and found to be effective in wound healing and maturation of the scar. Both preparations resulted in better wound healing when compared to the untreated control group and vaseline- treated group. This effect was comparable to that induced by MEBO, the positive control. CONCLUSION: The results indicate that A. fragrantissima has a pro-angiogenic effect, which may act through the VEGF signaling pathway.

The effect of green tea consumption on the expression of antioxidant- and inflammation-related genes induced by nicotine.

The aim of this study is to investigate the protective effect of green tea (GT) against the toxicity of nicotine. BALB/c mice were divided into four groups. Group I received food and water intake ad libidium, Group II received GT solution at a dose of 1 ml/kg body weight orally twice a day via gastric gavage, Group III was injected intraperitoneally with nicotine (2.5 mg/kg) once per day for 4 weeks, and Group IV received both nicotine and GT; GT was introduced using gastric gavage 1 hr before and 1 hr after the nicotine injection. The administration of nicotine altered the cellular antioxidant defense system by inducing inflammation and damage in the tissues of liver, lungs, and kidneys. In addition, nicotine treatment significantly enhanced the expression antioxidant- and inflammation-related genes. There were significant improvements when the nicotine-exposed mice treated with GT. PRACTICAL APPLICATIONS: In this study, it is revealed that the administration of nicotine altered the cellular antioxidant defense system by inducing inflammation manifested by the infiltration of inflammatory cells and damage seen in liver, lungs, and kidneys. GT contributed to the reduction of toxicity of nicotine, probably mediated by free radicals, through downregulation of nicotine-induced upregulated antioxidant- and inflammation-related genes. Never the less, further in depth investigation on characterization of the active constituents of GT responsible for their effect seen here and the mechanism that contributes to the effects seen in this reports is highly demanded. Furthermore, GT extract could be considered as a dietary supplement for the reduction of nicotine toxicity among cigarette smoker.

Correction to: The antiangiogenic activities of ethanolic crude extracts of four Salvia species.

CORRECTION: After the publication [1] it came to the attention of the authors that one of the co-authors was incorrectly included as Hamza Somrain. The correct spelling is as follows: Hamzeh Sumrein.

The antiangiogenic activities of ethanolic crude extracts of four Salvia species.

BACKGROUND: Angiogenesis is one of cancer hallmarks that are required for both cancer progression and metastasis. In this study we examined the antiangiogenic properties of the ethanolic crude extracts of four Salvia species grown in Jordan. METHODS: The direct antiangiogenic activity was evaluated using various models: ex vivo rat aortic ring assay, in vitro assessment of HUVEC proliferation and migration, and in vivo CAM assay, while we used the changes in the expression of HIF-1α and VEGF in breast cancer cells (MCF 7) as an indicative for the indirect antiangiogenic activity. RESULTS: All four crude extracts showed a potential antiangiogenic activity in the rat aortic assay, however two species were found to be cytotoxic against Fibroblast cell line (PLF); the finding that caused the exclusion of these two extracts from further studies. Of the two remaining extracts, S. triloba showed very promising direct and indirect antiangiogenic activities. S. triloba inhibited the HUVEC proliferation with an IC50 of 90 µg/mL and HUVEC migration by 82% at 150 µg/mL. Furthermore, the in vivo CAM assay also illustrated the high impact of S. triloba against the newly formed vessel in the chicken embryonic membrane. Interestingly, the S. triloba inhibited the expression of VEGF at the mRNA and protein and the HIF-1α mRNA in the MCF 7 breast cancer cells under both normoxic and hypoxic conditions. CONCLUSIONS: Taken together, all these findings of the direct and indirect angiogenic investigations nominated S. triloba as a highly potent antiangiogenic plant that may have chemotherapeutic and/or chemoprevention potentials.

ß-Caryophyllene causes regression of endometrial implants in a rat model of endometriosis without affecting fertility.

Many studies have shown that anti-inflammatory agents are effective in the treatment of endometriosis. ß-Caryophyllene exerted a potent anti-inflammatory effect in vivo. However, its effect on endometriosis has not been investigated. This study aims at investigating the effect of ß-caryophyllene on endometriosis and on fertility and reproduction in adult female rats. Autologous fragments of the endometrium were implantated in the peritoneal cavity in adult female rats. The growth of the endometriotic implants that developed after four weeks was recorded. Treatment started then with ß-caryophyllene (10 mg/kg or 30 mg/kg) or vehicle (control) for 21 days and the growth of the endometriotic implants was measured again. In fertility studies, female rats that received ß-caryophyllene or vehicle were mated and reproductive functions were observed including number and viability of implants, number of corpora lutea, length of pregnancy and outcome of litter. ß-Caryophyllene (10 mg/kg) suppressed the growth of endometriotic implants by 52.5% compared with controls. Also ß-caryophyllene produced apoptosis in luminal epithelim of the cyst as well as in endothelial cells of blood vessels. Ultrstructural studies revealed the presence of active mast cells and eosinophils in both control and ß-caryophyllene-treated rat cysts. No statistically significant difference was observed in any studied parameter between control and ß-caryophyllene-treated groups in fertility study. Therapy with ß-caryophyllene may present a promising novel, non-toxic therapeutic option for patients with endometriosis.

Thymoquinone efficiently inhibits the survival of EBV-infected B cells and alters EBV gene expression.

Epstein--Barr virus (EBV) is a human virus with oncogenic potentials that is implicated in various human diseases and malignancies. In this study, the modulator activity of the potent herbal extract drug thymoquinone on EBV was assessed in vitro. Thymoquinone was tested for cytotoxicity on human cells of lymphoblastoid cells, Raji Burkitt's lymphoma, DG-75 Burkitt's lymphoma, peripheral blood mononuclear cells, and periodontal ligament fibroblast. Apoptosis induction was analyzed via TUNEL assay and activity studies of caspase-3. The effect of thymoquinone on EBV gene expression was determined using real-time polymerase chain reaction. We report here, for the first time, a promising selective inhibitory affect of thymoquinone on EBV-infected B cell lines in vitro, compared with lower activity on EBV negative B cell line and very low toxicity on human peripheral blood mononuclear cells and periodontal ligament fibroblasts. Moreover, the drug was found to efficiently suppress the RNA expression of EBNA2, LMP1, and EBNA1 genes. Specifically, EBNA2 expression levels were the most affected indicating that this gene might have a major contribution to thymoquinone potency against EBV infected cells. Overall, our results suggest that thymoquinone has the potential to suppress the growth of EBV-infected B cells efficiently.

Screening the antiangiogenic activity of medicinal plants grown and sold in Jordan.

Angiogenesis is essential for the growth, invasion, and metastasis of most solid tumors and has become a valuable pharmacological target for cancer prevention and treatment. This study was performed to assess the antiangiogenic activity of 31 medicinal plants grown and sold in Jordan. The antiangiogenic activity was assessed using the rat aortic ring assay. Out of 31 extracts, 15 extracts showed more than 50 % inhibition of the blood vessels outgrowth from the primary tissue explants (p = 0.000). Three of these 15 extracts showed a potential cytotoxic effect on normal fibroblast cells. Four extracts shared antiangiogenic and antiproliferative activity towards MCF7 breast cancer cell lines. Eight extracts demonstrated selective antiangiogenic activity. This is the first report demonstrating the potential antiangiogenic activity of Artemisia judaica, Aloysia citriodora, Salvia egyptiaca, and Calendula arvensis. Some extracts with antiangiogenic activity exhibited selectivity against the endothelial cells proliferation, demonstrating a direct inhibitory activity against the key step in tumor angiogenesis.