Isolation of Cancer Stem Cells and Astrocytes from Human Glioblastoma: Morphological Characterization of Two Cells Types

Abstract  Background: Glioblastoma multiforme is the most aggressive astrocytoma in adults. Glioblastoma is a heterogenous tumor formed from various cells including astrocyte and cancer stem cells. Here, we explain the isolation, culture, morphology and specific markers of human glioblastoma astrocyte and stem cells. Methods: We isolated astrocyte and cancer stem cells from human glioblastoma tissue. The obtained glioblastoma tissues were digested and cultured in DMEM12, B27 supplemented with basic fibroblast growth factor and epidermal growth factor . The morphology and specific markers were assessed in astrocyte and cancer stem cell of human glioblastoma through immunochemistry. Results: Results indicated that there were two morphology types in cell culture including epithelioid morphology and fibroblastic morphology. The astrocyte confirmed via expression of the GFAP protein. Cancer stem cells were round and floating in the culture medium. Immunocytochemical staining indicated that nestin and SOX2 antigens were positively expressed in primary neurospheres. Conclusion: The expression of glial and stem cell markers show that both cells are in the human glioblastoma

The Synergistic Effect of Co-delivery of Anticancer Drugs Into Astrocytes Isolated From Human Glioblastoma Multiforme

Background: Chemotherapy drugs are not effective in the treatment of primary brain tumors due to the low efficacy of these drugs and drug transfer from the blood-brain barrier (BBB) toward the tumor site. Our purpose in this study was to assess the co-delivery of anticancer drugs to increase drug permeability from BBB.Methods: In this study, two chemotherapy drugs, namely methotrexate (MTX) and paclitaxel (PTX), were inserted into polyvinyl alcohol and poloxamer188-conjugated nanoparticles (NPs). Astrocytes were treated with different concentrations of 0-50 μg/ml from MTX, PTX, the MTX-PTX mixture, PTX-loaded NPs, MTX-loaded NPs, and PTX-MTX co-loaded NPs for 48 hours. The tumoricidal effect was assessed using the survival rate, Hoechst staining, and western blotting.Results: The results indicated significant reduction of the survival rate in astrocytes treated with PTX-MTX co-loaded NPs. In addition, apoptosis hallmarks consisting of fragmented DNA, overexpression of Bax, and expression reduction of Bcl-2 were in the cultured astrocytes.Conclusions: Our study proposes that the PTX-MTX co-delivery to NPs could be used as a possible approach for anti-cancer drug delivery to glioblastoma multiforme

Simulation of dose distribution and secondary particle production in proton therapy of brain tumor

AimThe aim of this study is simulation of the proton depth-dose distribution and dose evaluation of secondary particles in proton therapy of brain tumor using the GEANT4 and FLUKA Monte Carlo codes.BackgroundProton therapy is a treatment method for variety of tumors such as brain tumor. The most important feature of high energy proton beams is the energy deposition as a Bragg curve and the possibility of creating the spread out Bragg peak (SOBP) for full coverage of the tumor.Materials and methodsA spherical tumor with the radius of 1 cm in the brain is considered. A SNYDER head phantom has been irradiated with 30−130 MeV proton beam energy. A PMMA modulator wheel is used for covering the tumor. The simulations are performed using the GEANT4 and FLUKA codes.ResultsUsing a modulator wheel, the Spread Out Bragg Peak longitudinally and laterally covers the tumor. Flux and absorbed dose of secondary particles produced by nuclear interactions of protons with elements in the head are considerably small compared to protons.ConclusionsUsing 76.85 MeV proton beam and a modulator wheel, the tumor can be treated accurately in the 3-D, so that the distribution of proton dose in the surrounding tissues is very low. The results show that more than 99% of the total dose of secondary particles and protons is absorbed in the tumor

The effect of MDMA and pentoxifylline drug on bad/bcl-xl gene dosage expression changes on rat liver

        MDMA generally known as ecstasy, have deleterious effects on the serotonergic neurotransmitter system. Recent findings suggest that the liver and brain are major target organs of MDMA-related toxicities. Although most research is being dynamically performed on brain, however, the molecular mechanisms by which MDMA elicits adverse effects in both organs are poorly undrestood.The present study was performed to obtain evidence for molecular mechanism of apoptosis involved in MDMA-induced hepatotoxicity in rat liver after MDMAadministration. Moreover, the antagonistic effect of pentoxifylline was assessed on hepatotoxicity after MDMA administration. In this experimental study, sample size and power in each group were calculated as 10 rats with 95% confidence level and 5% confidence interval. In the study, four experimental groups were selected including Control Normal, MDMA, MDMA+PTX and PTX+MDMA. MDMA was dissolved in PBS and intraperitoneally injected three doses of 7.5mg/kg with two hours gap between doses. Pentoxyfilline also was injected as 100mg/kg, simultaneously with third dose of MDMA. After treatment, total RNA was isolated from liver tissue (5mg). Absorbance at 260nm, 280nm and 230nm were measured and immediately reverse transcription was performed. Included target genes were BAD and BCL-XL as pro-apoptotic and anti-apoptotic gene, respectively. After set up and validation, Real-Time PCR were performed and obtaining data were statistically analyzed to determine significantly differences between groups. Using Real-Time quantitative PCR results, BCL-XL gene expression ratio significantly increased in MDMA+PTX group. Moreover, BAD gene expression ratio increased and up-regulated in PTX+MDMA group (P-value <0.001).Our study focused on molecular mechanism of MDMA in programmed cell death using gene expression quantification of a pro-apoptotic and anti-apoptoic gene in MDMA-induced hepatotoxocity. The results shown MDMA prompted apoptosis in liver and pentoxifylline protects hepatotoxicity after and befor taking MDMA.

Gastrointestinal adverse effects of antiepileptic drugs in intractable epileptic patients

AbstractGastrointestinal (GI) discomforts are among the most common side effects of antiepileptic drugs (AEDs) that might lead to discontinuation or irregular consumption of the drugs. This study was conducted to evaluate the frequency of GI side effects of different AEDs in intractable epileptic patients treated with single or multiple drugs. GI discomfort of 100 epileptic patients (aged 35–76 years) treated with one or multiple AEDs was assessed. Seventy six patients (76%) were treated with two or more AEDs, and 24 (24%) were on monotherapy. The most common prescribed drug for monotherapy was carbamazepine and the most frequent combination was phenytoin and carbamazepine. Patients were suffering from different GI side effects including heartburn (34.6%), nausea (33.7%), constipation (26%), vomiting (22.1%), diarrhea (21.2%) and dysphagia (19.2%). Nausea and vomiting were significantly higher in patients receiving monotherapy with carbamazepine and valproic acid, respectively. When phenytoin, gabapentine, or valproic acid was added to the other AEDs, the risk of the occurrence of diarrhea, dysphagia, or heartburn was significantly increased, respectively. Addition of gabapentine to the other AEDs in multiple drug therapy was accompanied with the highest frequency of GI complications. This study indicated that GI side effects, which can affect drug absorption and utilization, were common in intractable epileptic patients with long-term AEDs treatment. This may influence the efficacy of the therapy with AEDs and enhance the probability of further attacks

Acute Transplantation of Human Olfactory Mucosa-Derived Olfactory Ensheathing Cells Fails to Improve Locomotor Recovery in Rats

Olfactory ensheathing cells-based therapy for spinal cord injury (SCI) repair has been a possible treatment for clinical study because of their safety in autologous transplantation and potential regenerative capability. However, there are contradictory reports on the results after transplantation in animal models. The purpose of this research was to investigate the effect of acute transplantation of human mucosa-derived olfactory ensheathing cells (OECs) on the repair of the spinal cord. Human olfactory ensheathing cells were isolated from the human mucosa and cultured under supplemented neuronal cell culture medium. They were characterized by immunocytochemistry for olfactory ensheathing cell markers. We induced spinal cord injury at T8-T9 of rats by aneurysm clips and simultaneously injected two million OECs into subarachnoid space of spinal cord. Sensory and motor behaviors were recorded by tail-flick reflex (TFR) and BBB scores, respectively every week for seven weeks after injury. Morphology and S100-beta antigen expression in olfactory ensheathing cells of the human olfactory mucosa was confirmed by immunostaining. OECs transplantation did not recover inflammation, neuronal vacuolation, hemorrhage, and cyst formation. These findings suggest that OECs transplantation in this experimental setting did not lead to tissue regeneration to enhance locomotion. These results broaden current knowledge and are additions to the science and literature

Novel Treatments for Alzheimer’s Disease

Alzheimer’s disease (AD) is a multifarious neurodegenerative disorder that leads to cognitive impairment and dementia in late adult life. Pathology hallmarks of Alzheimer’s disease were observed intracellular neurofibrillary tangles and extracellular amyloidal protein. According to pathology of this disease, abundant studies were performed with focused on pharmacological therapeutics over the last two decades. Current treatments for AD are acetylcholinesterase inhibitors (rivastigmine, galantamine, donepezil) and N-methyl D-aspartate receptor antagonist (memantine) that can decrease the progression of the disease. In this review, authors will discuss the various aspects of pathophysiological mechanisms and therapeutic strategies of Alzheimer’s disease

The Quality of Raw and Treated Water of Desalination Plants by Reverse Osmosis in Qeshm

Background: Desalination plants have an important role in providing community drinking water. The increasing trend of water consumption and decrease of natural resources of fresh water make this role even more important. The aim of this study was to evaluate the quality of inlet and outlet water of desalination plants by reverse osmosis (RO) process in Qeshm, Iran and also to compare the quality of outlet water from this process with National and International standards of drinking water. Methods: This study was performed by obtaining samples of inlet and outlet water of desalination plants working by reverse osmosis (RO) process in Qeshm for 7 months. Total hardness, electrical conductivity (EC), total dissolved solids (TDS), turbidity, temperature, pH, fluoride, nitrate, nitrite, chloride, sulfate and microbiological parameters were investigated. Data analysis was performed by SPSS16. Results: Total hardness and fluoride in the treated water from this process were lower than desirable limits and chloride concentration was higher than the permitted limit. The rest of the chemical and physical parameters were in acceptable range. Most qualitative parameters of the outlet water of the RO process decreased by 99% compared to the inlet water and outlet levels were in the standard range. Conclusion: Desalination plants by reverse osmosis (RO) process have a high efficiency in providing healthy drinking water based on qualitative standards. Keywords: Water quality, Water treatment, Desalination, Reverse osmosis proces

Cannabinoids as a Promising Therapeutic Approach for the Treatment of Glioblastoma Multiforme: A Literature Review

Gliobalstoma multiforme (GBM) or grade 4 astrocytoma is the most malignant form of primary brain tumor. Treatment of glioblastoma is difficult despite of surgery, radiotherapy and chemotherapy. Patients with glioblastoma survive for less than 12 months. Considering to biology function of glioblastoma, researchers have recently offered new therapeutic approaches such as cannabinoid therapy for glioblastoma. Cannabinoids are active compounds of Cannabis sativa that operate in the body similar to endogenous canabinoids –the endocannabinoids- through cell surface receptors. It is interesting that cannabinoids could exert a wide spectrum from antiproliferative effects in condition of the cell culture, animal models of glioblastoma and clinical trials. As a result, Cannabinoids seem to modulate intracellular signaling pathways and the endoplasmic reticulum stress response in glioma cells. Those play antitumoral effects through apoptosis induction and inhibition of glioblastoma angiogenesis. The goal of this study was to discuss cannabinoid therapy and also what cellular mechanisms are involved in the tumoricidal effect of the cannabinoids. In this review article, we will focus on cannabinoids, their receptor dependent functional roles against glioblastoma acccording to growth, angiogenesis, metastasis, and future purposes in exploring new possible therapeutic opportunities

The Neuroprotective Effect of Cannabinoid Receptor Agonist (WIN55,212-2) in Paraoxon Induced Neurotoxicity in PC12 Cells and N-methyl-D-aspartate Receptor Interaction

Objective: Considering that cannabinoids protect neurons against neurodegeneration, inthis study, the neuroprotective effect of WIN55,212-2 in paraoxon induced neurotoxicity inPC12 cells and the role of the N-methyl-D-aspartate (NMDA) receptor were evaluated.Materials and Methods: In this study PC12 cells were maintained in Dulbecco's modifiedeagle’s medium (DMEM+F12) culture medium supplemented with 10% fetal bovineserum. The cells were treated with paraoxon (200 μM) in the presence or absence ofWIN55,212-2 (0.1 μM), NMDA receptor agonist NMDA (100 μM), cannabinoid receptorantagonist AM251 and NMDA receptor antagonist MK801 (1 μM) at 15 minutes intervals.After 48 hours of exposure, cellular viability and protein expression of the CB1 receptorwere evaluated in PC12 cells.Results: Following the exposure of PC12 cells to paraoxon (200 μM), a reduction in cellsurvival and protein level of the CB1 receptor was observed (p<0.01). Treatment of thecells with WIN55,212-2 (0.1 μM) and NMDA (100 μM) prior to paraoxon exposure significantlyelevated cell survival and protein level of the CB1 receptor (p<0.01). Also, AM251(1μM) did not inhibit the cell survival and protein level of the CB1 receptor increase inducedby WIN55,212-2 (p<0.001). However, MK801 (1 μM) did inhibit cell survival andprotein expression of the CB1 receptor increase induced by NMDA (p<0.001).Conclusion: The results indicate that WIN55,212-2 and NMDA protect PC12 cellsagainst paraoxon induced toxicity. In addition, the neuroprotective effect of WIN55,212-2and NMDA was cannabinoid receptor-independent and NMDA receptor dependent, respectively