Iranian Journal of Basic Medical Sciences
Year:2016 | Volume:19 | Issue:10|Papers Count:15

Exosomes, as a mediator of cell-to-cell transfer of genetic information, act an important role in intercommunication between tumor cells and their niche including fibroblasts, endothelial cells, adipocytes and monocytes. Several studies have shown that tumor cells can influence their neighboring cells by releasing exosomes. These exosomes provide signaling cues for stimulation, activation, proliferation and differentiation of cells. Exosomes contain mRNAs, microRNAs (miRNA), and proteins that could be transferred to target cells inducing genetic and epigenetic changes. By facilitating the horizontal transfer of bioactive molecules such as proteins, RNAs and microRNAs, they are now thought to have vital roles in tumor invasion and metastases, inflammation, coagulation, and stem cell renewal and expansion. The aim of this review article is to discuss the significance of exosome-mediated intercellular communication within the tumor biology.

Objective(s): Particularly in developing countries, selenium and/or iodine deficiencies are encountered and use of pesticides in agriculture are not well-controlled. Fenvalerate is a pyrethroid insectide used in agriculture and has applications against a wide range of pests. This study was designed to evaluate the effects of fenvalerate on hepatic and cerebral xenobiotic metabolizing enzyme activities in the presence of iodine and/or selenium deficiency on a rat model. Materials and Methods: Iodine and/or selenium deficiency was induced by feeding three-week-old Wistar rats with a diet containing <0. 005 mg selenium kg-1, and/or administering 1% sodium perchlorate in drinking water for 7 weeks. Test groups received fenvalerate (100 mg kg-1 BW IP) for the last 7 days. Hepatic and cerebral microsomal aniline hydroxylase (CYP2E1) and cytosolic glutathione S-transferase (GST) activities were determined. Besides, hepatic NADPH-cytochrome P450 reductase (P450R), ethoxyresorufin O-deethylase (EROD, CYP1A1/1A2) and penthoxyresorufin O-depenthylase (PROD, CYP2B1/2B2), activities were also measured. Results: Fenvalerate had a general inductive effect on the hepatic and cerebral xenobiotic metabolizing enzyme activities. Moreover, enzyme activities were also altered by iodine and/or selenium deficiency, but the effects seemed to be enzyme-and tissue-specific. Conclusion: The inductive effect of fenvalerate, particularly in high dose exposures, may change the metabolism of several xenobiotics, including drugs, as well as endogenous substrates. The effects may vary depending on the selenium and/or iodine status of individual.

Objective(s): The purpose of the current study was to assess the feasibility of microspheres from biocompatible polymer for oral bioavailability (BA) enhancement of potent sulfonamide-type loop diuretic-Furosemide-which used in the treatment of congestive heart failure, caused edema, cirrhosis, renal disease and as an adjunct in acute pulmonary edema. The comparatively poor and inconstant BA of furosemide, which occurs site-specifically in the stomach and upper small intestine, has been ascribed to the poor dissolution of furosemide. Materials and Methods: In attempt to enhance the drug BA, poly (dl-lactic-co-glycolic acid) (PLGA) microspheres of furosemide were obtained using solvent-evaporation method and the carrier characteristics were investigated subsequently. Results: The in vivo performance of optimum formulation was assessed by pharmacokinetic evaluation of drug after orally administration of free and loaded in microspheres to rats (4 mg/Kg). For this reason, the concentration of drug in plasma was measured by a new developed and sensitive method of HPLC. Acceptable drug loading and encapsulation efficiency of microspheres were obtained to be 70. 43 and 85. 21 %, respectively. Microspheres provided improved pharmacokinetic parameters (Cmax = 147. 94 ng/ml, Tmax = 1. 92 hr) in rats as compared with pure drug (Cmax = 75. 69 ng/ml, Tmax = 1. 5 hr). The obtained AUC of drug in microsphere was 10 fold higher than of the free drug. Conclusion: The results showed that the prepared microspheres successfully improved BA of the poorly water-soluble drug effectively.

Objective(s): This study describes the preparation, biodistribution of 153Sm-DTPA-SPION after intravenous injection in rats. Materials and Methods: The chelator DTPA dianhydride was conjugated to SPION using a small modification of the well-known cyclic anhydride method. Conjugation was done at a 1: 4 (SPION: ccDTPA) molar ratio. Conjugation reaction was purified with magnetic assorting column (MACs) using high gradient magnetic field following incubation, the radio labeled conjugate was checked using RTLC method for labeling and purity checked. Results: The RTLC showed that labeling yield was above 99% after purification and the compound have good in vitro stabilities until 48 hr post injection in the presence of human serum. The biodistribution of 153Sm-DTPA-SPION in rats showed dramatic uptake in the reticuloendothelial system (RES) and their clearance is so fast in other organs especially in the blood. Biodistribution results show that after 30 min post injection more than 84% of injected activities were taken up by the liver and spleen (about 64% and 20%, respectively). Conclusion: Due to magnificent uptakes of this radiotracer in the liver and spleen and their fast clearance from other tissues, especially in blood, it is suggested that this radiotracer would be a potential candidate for RES theranostic purposes.

Objective(s): β-thalassemia is one of the most common genetic disorders in the world. As one of the promising treatment strategies, fetal hemoglobin (Hb F) can be induced. The present study was an attempt to reactivate the γ-globin gene by introducing a gene construct containing KLF1 binding sites to the K562 cell line. Materials and Methods: A plasmid containing a 192 bp sequence with two repeats of KLF1 binding sites on β-globin and BCL11A promoters was constructed and used to transfect the K562 cell line. Positive selection was performed under treatment with 150 μ g/ml hygromycin B. The remaining cells were expanded and harvested on day 28, and genomic DNA was extracted. The PCR was carried out to verify insertion of DNA fragment to the genome of K562 cells. The cells were differentiated with 15 μ g/ml cisplatin. Flowcytometry was performed to identify erythroid differentiation by detection of CD235a+ cells. Real-time RT-PCR was performed to evaluate γ-globin expression in the transfected cells. Results: A 1700 bp fragment was observed on agarose gel as expected and insertion of DNA fragment to the genome of K562 cells was verified. Totally, 84% of cells were differentiated. The transfected cells significantly increased γ-globin expression after differentiation compared to untransfected ones. Conclusion: The findings demonstrate that the spongy effect of KLF1-binding site on BCL11A and β-globin promoters can induce γ-globin expression in K562 cells. This novel strategy can be promising for the treatment of β-thalassemia and sickle cell disease.

Objective(s): Long-term treatment with antipsychotics causes serious side effects such as tardive dyskinesia that characterized by abnormal movements in the orofacial region. Oxidative stress in the brain specific area is implicated in the pathophysiology of tardive dyskinesia. In this study the protective effect of crocin on haloperidol-induced orofacial dyskinesia was evaluated. Materials and Methods: Haloperidol (1 mg/kg, IP) and crocin (10, 20 and 40 mg/kg, IP) were administrated to rats for 21 days. Behavioral assessments such as orofacial dyskinesia movements, open field test and elevated plus maze (EPM) were evaluated every week. Malondealdehyde (MDA) and glutathione (GSH) levels in the hippocampus, cortex and striatum were also measured. Results: Haloperidol increased vacuous chewing movements (VCMs) and tongue protrusions (TPs) in rats and co-administration of crocin (20 and 40 mg/kg) significantly reduced them. Furthermore, haloperidol decreased the locomotor and exploratory activities (rearing) in the open field test and decreased the percentage of entries into open arms and the percentage of the time spent on open arms in the EPM. Pretreatment with crocin (10 mg/kg) modified haloperidol effects on these behavioral parameters. Haloperidol induced lipid peroxidation in three brain regions, whereas crocin co-administration reduced the MDA and restored the decreased GSH levels. Conclusion: Our finding suggests that oxidative stress has an important role in the development of tardive dyskinesia. Crocin showed protective effect against haloperidol induced tardive dyskinesia and as a potent naturally antioxidant could be a new and useful drug and a possible therapeutic option for the treatment of tardive dyskinesia.

Objective(s): Neuroprotective effects of female gonadal steroids are mediated through several pathways involving multiple peptides and receptors after traumatic brain injury (TBI). Two of these peptides are including the regulatory peptides neuromedin U (NMU) and neuromedin S (NMS), and their common receptor neuromedin U2 receptor (NMUR2). This study investigates the effects of physiological doses of estradiol and progesterone on brain edema, NMS and NMU as well as NMUR2 expression following TBI. Materials and Methods: Ovariectomized female rats were given high-and low-dose of female sex steroid hormones through implantation of capsules for a week before trauma. The brain NMUR2 expression, prepro-NMS expression, NMU content, and water content (brain edema) were evaluated 24 hr after TBI induced by Marmarou’ s method. Results: Percentage of brain water content in high-and low-dose estradiol, and in high-and low-dose progesterone was less than vehicle (P<0. 01). Results show high expression of prepro-NMS in high dose progesterone (TBI-HP) rats compared to the high dose estrogen (TBI-HE), as well as vehicle (P<0. 01). NMU content in low-dose progesterone (TBI-LP) group was more than that of vehicle group (P<0. 001). Furthermore a difference in NMU content observed between TBI-HP compared to TBI-HE, and vehicle (P<0. 05). The NMUR2 mRNA expression revealed an upregulation in TBI-HP rats compared to the TBI-HE group (P<0. 001). Conclusion: Findings indicate that progesterone attenuates brain edema and induces an increase in NMS and its receptor which may mediate the anti-edematous effect of progesterone after TBI.

Objective(s): Evaluation of the Sophora alopecuroides var. alopecuroides seed effects on morphine withdrawal syndrome in mice and determination of the alkaloid composition of the seed total extract. Materials and Methods: The effects of the seed total extract, alkaloid fraction and major compound matrine on the mice morphine withdrawal syndrome were compared to saline and methadone. Mice were made dependent on morphine by morphine sulfate injection 3 times a day for 3 days. The withdrawal jumping and diarrhea were induced by administration of naloxone 2 hr after the 10th injection of morphine sulfate on the day 4. The total extract (100, 200, 300 mg/kg), alkaloid fraction (5, 10, 20 mg/kg), matrine (5, 15, 30 mg/kg), methadone (10 mg/kg) or saline were injected 30 min before naloxone. All drugs were administered by subcutaneous injection. The total extract alkaloid composition was also determined by gas chromatography (GC) and GC-MS analysis. Results: All doses of the total extract, alkaloid fraction and matrine as well as methadone decreased jumping and diarrhea significantly compared to the saline. The effects of the total extract and alkaloid fraction were not significantly different from methadone. But, there were significant differences between the effects of matrine and methadone. Matrine, cytisine, sophoridine, n-methyl cytisine, sophocarpine and sophoramine were the major alkaloids. There was no nicotine in the total extract. Conclusion: S. alopecuroides var. alopecuroides suppresses opioid withdrawal with efficacy comparable to methadone. Matrine may be one of the alkaloids responsible for the effect of the plant.

Objective(s): In recent years, polypropyleneimine (PPI) dendrimers have attracted great interest as nonviral gene delivery systems because of their attractive features including highly branched architecture with number of reactive end groups. However, without being structurally modified, they are not efficient gene carriers. In the present study, generation 2 and 3 (G2 and G3) of PPI dendrimers were conjugated with alkylcarboxylate groups as linker to enhance the transfection efficiency while maintaining their low cell toxicity. Materials and Methods: First, 10-bromodecanoic acid was covalently attached to all available surface primary amines of PPI G2 and G3 to increase their lipophilicity. In the subsequent step, PPIs were conjugated to the alkylcarboxylate groups of alkylcarboxylate-PPI derivatives to increase the number of surface primary amines. Physicochemical properties of modified PPIs were determined. Transfection experiments (using both luciferase and green fluorescent protein (GFP)-expressing plasmids) and cytotoxicity assay were performed to evaluate the efficiency of the final derivatives. Results: Fabricated vectors condensed DNA effectively so that polyplexes with appropriate size (below 155 nm) and positive surface charge were constructed. Cross-linked low molecular weight PPIs (G2 or G3) with decanoate linkage increased transfection efficiency significantly while maintaining the low cytotoxicity. PPI G2 derivative exhibited increased buffering capacity which is believed to be responsible for better proton sponge mechanism leading to higher transfection efficiency. Conclusion: Our results indicated that oligomerization of low molecular weight PPI (PPI G2-alkyl-PPI G2 conjugate) could be an approach to increase the transfection efficiency and to lower the cytotoxicity of low molecular weight polycations.

Objective(s): The periodic binding of protein expressed by Mycobacterium tuberculosis H37Rv with the host cell receptor molecules i. e. fibronectin (Fn) is gaining significance because of its adhesive properties. The genome sequencing of M. tuberculosis H37Rv revealed that the proline-glutamic (PE) proteins contain polymorphic GC-rich repetitive sequences (PGRS) which have clinical importance in pathogenesis events when the host encounters M. tuberculosis H37Rv. The functional parts of PE_PGRS family proteins, have not been extensively studied in tuberculosis biology. Materials and Methods: Fibronectin (10 ng and 20 ng) were used for FnBP assay and its enzymatic activities were observed by using various protein concentrations. Results: Therefore, in the present work, we cloned, expressed, purified and identified a novel PE_PGRS61 (Rv3653) family protein in M. tuberculosis H37Rv. Our experiment, observation suggested that at particular concentrations of 10 ng and 20 ng of Fn exhibits optimum binding to the purified Fibronectin Binding Protein (FnBP), a PE_PGRS61 family protein at 0. 20 μ g and 0. 25 μ g concentrations, respectively. Moreover, for better understanding the computational analysis, the B-cell and T-cell epitopes prediction prospect some amino acid propensity scales with hydrophilicity and antigenic variation index at their respective locations. Conclusion: Thus, the current findings provide an opportunity to illuminate the functions of PE_PGRS61 family protein. So, in this point of view, it could be useful to develop a novel therapeutic approach or diagnostic pipeline through targeting these fibronectin binding protein (FnBP) expressing genes.

Objective(s): Bidens species are used for their antidiabetic properties traditionally in many countries. Aim of this study is to evaluate hypoglycaemic and antidiabetic activity of Bidens tripartita extract and to identify its active compounds through bioactivity guided isolation technique. Materials and Methods: Hypoglycaemic effects of B. tripartita extract and its sub-extracts were investigated in normal and glucose-hyperglycaemic rats. Streptozotocin induced diabetic rats were used to examine antidiabetic activity of the extract and its sub-extracts after acute and sub-acute administration. Additionally, in vitro enzyme inhibitory and antioxidant activities were evaluated. HPLC analyses were carried out to determine the active constituents of the extract and its sub-extracts. Results: Through in vivo bioactivity-guided fractionation process, ethyl acetate and n-buthanol sub-extracts were found to have potent antidiabetic activity. In vitro enzyme inhibitory activities of the same sub-extracts were found to be potent. The highest total phenol, flavonoid contents and radical scavenging activity was determined in ethyl acetate sub-extract. According to LC-MS analyses, chlorogenic acid, luteolin and 7-O-glucoside of luteolin (cynaroside) were determined as the main components of the active sub-extracts. Conclusion: According to our results, B. tripartita has potent antidiabetic activity and its active constituents might be beneficial for diabetes and its complications.

Objective(s): Sugar cane molasses is a commonly used ingredient in several food products. Contrasting reports suggest that molasses may have potential adverse or beneficial effects on human health. However, little evidence exists that examines the effects of molasses on the different physiological systems. This study investigated the effects of sugar cane molasses on various physiological systems using in vivo and in vitro methods. Materials and Methods: Molasses was administered orally to BALB/c, male mice and animals were randomly assigned into either a treatment or control group. General physiological changes, body weight and molasses intake of animals were monitored. At the end of the exposure period, collected blood samples were evaluated for potential toxicity using plasma biomarkers and liver enzyme activity. Immunised treated and untreated mice were evaluated for antibody titre to determine the effect of molasses on the immune response. To investigate the impact of molasses on testicular steroidogenesis, testes from both treated and control groups were harvested, cultured and assayed for testosterone synthesis. Results: Findings suggest that fluid intake by molasses-treated animals was significantly increased and these animals showed symptoms of loose faeces. Molasses had no significant effect on body weight, serum biomarkers or liver enzyme activity (P>0. 05). Immunoglobulin-gamma anti-antigen levels were significantly suppressed in molasses-treated groups (P=0. 004). Animals subjected to molasses exposure also exhibited elevated levels of testosterone synthesis (P=0. 001). Conclusion: Findings suggests that molasses adversely affects the humoral immune response. The results also promote the use of molasses as a supplement to increase testosterone levels.

Objective(s): Similar characteristics of molecular pathways between cellular reprogramming events and tumorigenesis have been accentuated in recent years. Reprogramming-related transcription factors, also known as Yamanaka factors (OCT4, SOX2, KLF4, and c-MYC), are also well-known oncogenes promoting cancer initiation, progression, and cellular transformation into cancer stem cells. Long non-coding RNAs (lncRNAs) are a major class of RNA molecules with emerging roles in stem cell pluripotency, cellular reprogramming, cellular transformation, and tumorigenesis. The long intergenic non-coding RNA ROR (lincRNA-ROR, linc-ROR) acts as a regulator of cellular reprograming through sponging miR-145 that normally negatively regulates the expression of the stemness factors NANOG, OCT4, and SOX2. Materials and Methods: Here, we employed a real-time PCR approach to determine the expression patterns of linc-ROR and its two novel spliced variants (variants 2 and 4) in esophageal squamous cell carcinoma (ESCC). Results: The quantitative real-time RT-PCR results revealed a significant up-regulation of linc-ROR (P=0. 0098) and its variants 2 (P=0. 0250) and 4 (P=0. 0002) in tumor samples of ESCC, compared to their matched non-tumor tissues obtained from the margin of same tumors. Our data also demonstrated a significant up-regulation of variant 4 in high-grade tumor samples, in comparison to the low-grade ones (P=0. 04). Moreover, the ROC curve analysis demonstrated that the variant 4 of ROR has a potential to discriminate between tumor and non-tumor samples (AUC=0. 66, P<0. 05). Conclusion: Our data suggest a significant up-regulation of linc-ROR and its variants 2 and 4 in ESCC tissue samples.

Objective(s): This study aims to determine the effects of ozone therapy on restoring impaired Nrf2 activation to ameliorate chronic tubulointerstitial injury in rats with adenine-induced CKD. Materials and Methods: Sprague– Dawley rats were fed with 0. 75% adenine-containing diet to induce CKD and chronic tubulointerstitial injury. Ozone therapy was administered by rectal insufflation. After 4 weeks, serum and kidney samples were collected and analyzed. Renal function and systemic electrolyte level were detected. Pathological changes in kidney were assessed by hematoxylin– eosin staining and Masson trichrome staining. Nrf2 activation was detected by immunohistochemistry and Western blot analyses. The levels of SOD, CAT, GSH, PCO, and MDA were detected in the kidney. Immunohistochemistry, Western blot, and real-time PCR analyses were performed to evaluate the activation of the nuclear factor kappa B (NF-κ B) P65 pathway and inflammation infiltration in the tubulointerstitium of the rats. Results: Ozone therapy improved severe renal insufficiency and tubulointerstitial morphology injury as well as restored Nrf2 activation and inhibited the NF-κ B pathway in rats with adenine-induced CKD. Ozone therapy also up-regulated anti-oxidation enzymes (SOD, CAT, and GSH) and down-regulated oxidation products (PCO and MDA), as well as inflammatory cytokines (IL-1β , IL-6, TNF-α , and ICAM-1) in the kidney. Conclusion: These findings indicated that ozone therapy could attenuate tubulointerstitial injury in rats with adenine-induced CKD by mediating Nrf2 and NF-κ B.

Objective(s): The aim of the present study was to evaluate the effect of Quercus brantii galls extract on the rat skin burn wound healing. Materials and Methods: Ethanol extract of the galls of Q. brantii was used to treat the induced burn wounds on the back of 32 Wistar rats divided into 4 groups. The groups were treated by placebo, 1%, 2% and 4% concentration gall extract gels for 14 days and the efficacy of treatment was assessed based on reduction of burn wound area, as well as histological and molecular characteristics. Results: The mean wound surface in the 14th day, in all groups treated by Q. brantii gall extracts were larger than control group and the differences were statistically significant (P=0. 043). The mean histological wound healing scores were not statistically different. Analysis of nitric oxide and platelet derived growth factor concentration in wound fluids in the 5th day of study showed that there was not any significant difference between groups (P=0. 468 and 0. 312 respectively). Fibroblast growth factor (bFGF) concentration in the wound fluids, was significantly higher in group treated with 1% gall extract gel in comparison to the control group (P=0. 026). Conclusion: Our results could not prove the significant positive effect of Q. brantii galls extract on the burning wound healing. More studies with more groups treated with different doses of the Q. brantii extract are recommended.