Ferutinin, an apoptosis inducing terpenoid from Ferula ovina.

A current hurdle in cancer management is the intrinsic or acquired resistance of cancer cells to chemical agents that restricts the efficacy of therapeutic strategies. Accordingly, there is an increasing desire to discover new natural compounds with selective toxicity to combat malignancies. In present study, the cytotoxic and apoptosis- inducing activities of ferutinin, a terpenoid derivative from Ferula ovina, were investigated on human breast (MCF7) and bladder (TCC) cancer cells as well as normal fibroblasts (HFF3).The toxicity and DNA damage inducing effects of ferutinin were studied by MTT and comet assays, DAPI and PI staining and DNA laddering. The IC50 values of ferutinin were identified and compared with routine prescribed drugs, doxorubicin and vincristine, by MTT test. Alkaline comet assay and DAPI staining revealed DNA damage due to ferutinin, which was significantly (p<0.001) higher in MCF7 and TCC than HFF3 cells. Apoptosis induction was evidenced by PI staining and DNA laddering. Our results suggest that ferutinin could be considered as an effective anticancer agent for future in vivo and clinical experiments.

The Effect of Ciprofloxacin Injection on Genetically Absence Prone (Wag/Rij) Rat's Electroencephalogram Characteristics.

INTRODUCTION: Ciprofloxacin which was used in this study is a Fluoroquinolone (FQ). This kind of drug may cause epileptic seizures probably because of the inhibition of GABA binding to its receptors. Wag/Rij rats (an animal model for generalized absence epilepsy), were used as experimental subjects. METHODS: For EEG study, electrodes were inserted into the cortex of animals according to paxinos coordinates. After and before ciprofloxacin injection, EEG was recorded and their SWDs were compared with each others. RESULTS: Findings showed a significant increase in the mean number of seizures during recording period. But the mean number of SWDs during seizures did not show any significant differences between groups. DISCUSSION: These results may be due to involvement of GABA antagonistic effects of FQs and/or Mg2+ linked blockade of NMDA receptors. More researches are going to determine physiopathology of SWDs and find new effective substance against this kind of epilepsy.

Investigating the cytotoxic and apoptosis inducing effects of monoterpenoid stylosin in vitro.

The aim of this study was to investigate the cytotoxic and anticancer activities of stylosin, a monoterpene extracted from an edible plant, Ferula ovina, on 5637 and HFF3 cells using MTT and comet assays and DAPI staining. To assess stylosin effects, cells were cultured in the presence of various concentrations of stylosin during three days; the IC(50) of stylosin on cancerous 5637 cells was less than its value on HFF3 normal cells, indicating that it might have anticancer properties. Investigating the mechanism of stylosin action revealed that it quickly induced DNA lesions and increased the number of apoptotic cells.

Investigating the enhancement of cisplatin cytotoxicity on 5637 cells by combination with mogoltacin.

Transitional cell carcinomas (TCCs), which account for 90% of bladder cancers, arise from the transitional epithelium of bladder. Cisplatin is a chemotherapeutic drug used to treat bladder cancer, but intrinsic and acquired resistance to cisplatin limit its effectiveness. The aim of this study was to determine the ability of mogoltacin, a sesquiterpene-coumarin from Ferula badrakema, to enhance cytotoxic effects of cisplatin on 5637 cells, using MTT assay, comet method, DAPI staining and efflux assay. In order to analyse mogoltacin combinatorial effects, 5637 cells were cultured in the presence of various combined concentrations of mogoltacin and cisplatin. The results of MTT assay revealed that combination of 1 µg/mL cisplatin+32 µg/mL mogoltacin, increased the cytotoxicity of cisplatin by 45.3%. Investigating the mechanism of this action by comet assay indicated that mogoltacin increases the apoptotic effects of cisplatin on 5637 cells via DNA lesion by 44%. Furthermore, studying nuclear morphological changes revealed that the combination of mogoltacin+cisplatin significantly (P<0.001) increases the number of apoptotic cells. Results of efflux assay indicated that mogoltacin did not have any significant effect on the activity of MDR transporters, therefore, this sesquiterpene-coumarin increases the effects of cisplatin possibly by interacting with other drug transporters.

The beneficial effects of olibanum on memory deficit induced by hypothyroidism in adult rats tested in Morris water maze.

Functional consequences of hypothyroidism include impaired learning and memory and inability to produce long-term potentiation (LTP) in hippocampus. Olibanum has been used for variety of therapeutic purposes. In traditional medicine, oilbanum is used to enhance learning and memory. In the present study the effect of olibanum on memory deficit in hypothyroid rats was investigated. Male wistar rats were divided into four groups and treated for 180 days. Group 1 received tap drinking water while in group 2, 0.03% methimazol was added to drinking water. Group 3 and 4 were treated with 0.03% methimazole as well as 100 and 500 mg/kg olibanum respectively. The animals were tested in Morris water maze. The swimming speed was significantly lower and the distance and time latency were higher in group 2 compared with group 1. In groups 3 and 4 the swimming speed was significantly higher while, the length of the swim path and time latency were significantly lower in comparison with group 2. It is concluded that methimazole-induced hypothyroidism impairs learning and memory in adult rats which could be prevented by using olibanum.

Gene silencing in human embryonic stem cells by RNA interference.

RNA interference (RNAi) is a post-transcriptional conserved mechanism, which is present in a wide range of organisms and leads to specific gene silencing. The effector molecules in this process are double-stranded RNAs (dsRNAs) that are homologous in sequence to the silenced gene and are processed into small interfering RNAs (siRNAs) by an enzyme called Dicer. Consequently, siRNAs are incorporated into an RNA-induced silencing complex, which finds and cleaves the target mRNA. Because of its exquisite specificity and efficiency, RNAi is being considered as an important tool for gene silencing in living organisms. Human embryonic stem cells (HESCs) have the ability to both self-renew and differentiate into cell types of all three germ layers. HESCs open new avenues for understanding some of the very early lineage determination events that occur during embryogenesis and are also considered as an important source of cells for cell replacement therapies. Understanding how the signaling pathways orchestrate and direct HESC differentiation toward certain cell types is critical for basic research. In this way RNAi appears to be a valuable tool in stem cell biology for dissecting the pathways involved in differentiation, lineage segregation and production of cells for cell therapy. Here, we review the prospects of combining RNAi and HESC manipulation for both basic research and future therapies.

Cytotoxicity of vincristine on the 5637 cell line is enhanced by combination with conferone.

Bladder cancer is one of the most common cancers worldwide, with the highest incidence in industrialized countries. There are three major histological subtypes of bladder cancer: transitional cell carcinoma (TCC) (> 90%), squamous cell carcinoma (< 10%) and adenocarcinoma (1-2%). The present study was carried out to assess the effects of conferone, a sesquiterpene coumarin isolated from Ferula badrakema, on a TCC subline, 5637 cells. In order to test the effects of conferone, 5637 cells were treated with different concentrations (16, 32, 64, 128 microg/ml) of conferone. The results indicated that conferone did not have any significant cytotoxic effect on these neoplastic cells. To determine the combining effects, the cells were cultured in the presence of different concentrations of conferone (16, 32, 64, 128 microg/ ml) and vincristine (30, 40, 50 microg/ml) in combination. The morphological changes were then observed and cytotoxicity effects were studied using the MTT assay 24, 48 and 72 h following drug administration. The cells were more rounded and granulated after treatments with both drugs in comparison to vincristine only. The results of the MTT assay confirmed the morphological observations. After 48 h of combined treatment with 40 microg/ml vincristine and 16 microg/ml conferone, the cytotoxicity of vincristine was increased by 23.6%.

Effect of thymoquinone on ethylene glycol-induced kidney calculi in rats.

INTRODUCTION: The aim of this study was to investigate the effects of thymoquinone, a major component of Nigella Sativa seeds on ethylene glycol-induced kidney calculi in rats. MATERIALS AND METHODS: Sixty male Wistar rats were randomly divided into 6 groups (intact control, ethylene glycol control, and 4 experimental groups) and treated for 28 days according to the protocol of the study. The rats in experimental groups received ethylene glycol and intraperitoneal injection of thymoquinone either from the first day of the study or the 15th day, with either doses of 5 mg/kg or 10 mg/kg. Blood and 24-hour urine samples were collected at baseline and on day 28. Urine oxalate and citrate and serum electrolytes were also measured. On day 29, all rats were decapitated and their kidney specimens were studied. RESULTS: On day 28, urine oxalate concentration significantly decreased in the experimental groups compared to the ethylene glycol group (P < .001). Also, serum calcium levels were significantly higher in the experimental groups (P = .001). Calcium oxalate deposits were smaller in the experimental groups than the ethylene glycol group. The mean number of deposits was lower in these groups, too (P < .001). Treatment with the lower dose of thymoquinone was associated with fewer deposits. CONCLUSION: Thymoquinone significantly decreased the number and size of calcium oxalate deposits in the renal tubules. The dose and duration of treatment, however, does not have a linear relation with the outcomes. Further studies on thymoquinone as a preventive and therapeutic drug for kidney calculi are suggested.