Iranian Journal of Basic Medical Sciences
Year:2015 | Volume:18 | Issue:7|Papers Count:14

Objective (s): Human cytomegalovirus (CMV), a double‐strand DNA herpesvirus, can be transmitted via blood transfusion which is especially important for immunocompromised recipients and can cause a fatal infection. CMV seroprevalence in Iran was studied on blood donors, healthy subjects, and some patients. Highly variable rates were detected. The purpose of this study was to review CMV seroprevalence in blood donors and apparently healthy individuals, in Iran.Materials and Methods: One hundred and fifty‐eight electronic and paper‐based resources and databases including published articles in internal and external journals, seminars, dissertations, and theses available in the database and different websites were used to be systematically reviewed as a meta‐analysis. Less related articles to the issue, papers of specific high risk population, and articles with not enough information, were excluded. Eventually 22 articles that satisfied our selection criteria were systematically reviewed and analyzed. To explore heterogeneity between studies the I square (I2) index was used. Data were analyzed using the statistical software package (STATA) 11.Results: The heterogeneity between selected studies was 97% with an I2 statistic. In this study a random effects model was used for meta‐analysis. The prevalence of CMV IgG and CMV IgM antibodies in the country were estimated to be 92% (95% CI: 90‐94) and 2.6% (95% CI: 1.7‐3.6), respectively.Conclusion: Given high rate of CMV seropositivity in Iran, it seems that CMVAbs screening would not be a reasonable and affordable approach to prevent CMV infection via transfusion especially for immune compromised recipients, so alternative strategies should be considered.

Objective (s): The colorectal cancer stem cells (CSCs) with the CD133+phenotype are a rare fraction of cancer cells with the ability of self‐renewal, unlimited proliferation and resistance to treatment.Quercetin has anticancer effects with the advantage of exhibiting low side effects. Therefore, we evaluated the anticancer effects of quercetin and doxorubicin (Dox) in HT29 cancer cells and its isolated CD133+CSCs.Materials and Methods: The CSCs from HT29 cells were isolated using CD133 antibody conjugated to magnetic beads by MACS. Anticancer effects of quercetin and Dox alone and in combination on HT29 cells and CSCs were evaluated using MTT cytotoxicity assay and flow cytometry analysis of cell cycle distribution and apoptosis induction.Results: The CD133+CSCs comprised about 10% of HT29 cells. Quercetin and Dox alone and in combination inhibited cell proliferation and induced apoptosis in HT29 cells and to a lesser extent in CSCs. Quercetin enhanced cytotoxicity and apoptosis induction of Dox at low concentration in both cell populations. Quercetin and Dox and their combination induced G2/M arrest in the HT29 cells and to a lesser extent in CSCs.Conclusion: The CSCs were a minor population with a significantly high level of drug resistance within the HT29 cancer cells. Quercetin alone exhibited significant cytotoxic effects on HT29 cells and also increased cytoxicity of Dox in combination therapy. Altogether, our data showed that adding quercetin to Dox chemotherapy is an effective strategy for treatment of both CSCs and bulk tumor cells.

Objective (s): Silver nanoparticles (Ag‐NPs) are used widely in bedding, water purification, tooth paste and toys. These nanoparticles can enter into the body and move into the hippocampus. The aim of this study was to investigate the neurotoxicity of silver nanoparticles in the adult rat hippocampus.Materials and Methods: 12 male Wistar rats were randomly divided into two experimental and control groups (6 rats in each group). Animals in the experimental group received Ag‐NPs (30 mg/kg) orally (gavage) for 28 consecutive days. Control group in the same period was treated with distilled water via gavage. At the end of experiment, animals were deeply anesthetized, sacrificed, and their brains were collected from each group. Finally the brain sections were stained using toluidine blue and TUNEL.Then to compare the groups, dark neurons (DNs) and apoptotic neurons were counted by morphometric method.Results: Results showed that the numbers of DNs and apoptotic cells in the CA1, CA2, CA3, and dentate gyrus (DG) of hippocampus significantly increased in the Ag‐NPs group in comparison to the control group (P<0.05).Conclusion: Exposure to Ag‐NPs can induce dark neuron and apoptotic cells in the hippocampus.

Objective (s): Phenylketonuria (PKU) is a genetic inborn error of phenylalanine (Phe) metabolism resulting from insufficiency in the hepatic enzyme, phenylalanine hydroxylase (PAH), which leads to elevated levels of Phe in the blood. The present study was carried out for mutation analysis of the PAH gene in West Azerbaijan province of Iran.Materials and Methods: A total of 218 alleles from 40 PKU families were studied using restriction fragment length polymorphism‐polymerase chain reaction (RFLP‐PCR) method.Results: The frequencies of IVS10‐11, S67P, R261Q, R252W, IVS11nt‐1 g>c, R408Q, and Q232Q mutations were 28 (35), 17 (21.25), 15 (18.75), 3 (3.75), 3 (3.75), 2 (2.5), and 1 (1.25), in cases group, and 51 (23.4), 31 (14.2), 27 (12.4), 6 (2.75), 6 (2.75), 4 (1.83), and 2 (0.92) in total group, respectively. The mutations of R243Q, 364delG, L333F, 261X, I65T, and R408W were not detected in our samples.Conclusion: It can be concluded that the IVS10‐11 mutation has the highest frequency in the tested population. To our knowledge, this report is the first in its own kind and provides better understanding of the genetic heterogeneity, the origin and distributions of PAH mutations in West Azerbaijan province of Iran.

Objective (s): Recently anti‐inflammatory effects of antidepressants have been demonstrated.Venlafaxine belongs to newer antidepressants with serotonin norepinephrine reuptake inhibition property. The pain alleviating properties of venlafaxine in different pain models such as neurogenic pain, diabetic neuropathy, and fibromyalgia have been demonstrated. Anti‐inflammatory effects of venlafaxine and also its underlying mechanisms remain unclear. The present study was designed to evaluate the anti‐inflammatory effects of venlafaxine and determine possible underlying mechanisms.Materials and Methods: We examined the anti‐inflammatory effects of intraperitoneal (IP) and intracerebroventricular (ICV) administration of venlafaxine in the rat model of carrageenan‐induced paw edema.Results: Our results showed that both IP (50 and 100 mg/kg) and ICV (50 and 100 mg/rat) injection of venlafaxine inhibited carrageenan‐induced paw edema. Also IP and ICV administration of venlafaxine significantly decreased myeloperoxidase (MPO) activity and interleukin (IL) ‐1b and tumor necrosis factor (TNF) ‐a production. Finally, we tried to reverse the anti‐inflammatory effect of venlafaxine by yohimbine (5 mg/kg, IP), an alpha2‐adrenergic antagonist. Our results showed that applied antagonist failed to change the anti‐inflammatory effect of venlafaxine.Conclusion: These results demonstrated that venlafaxine has potent anti‐inflammatory effect which is related to the peripheral and central effects of this drug. Also we have shown that anti‐inflammatory effect of venlafaxine is mediated mostly through the inhibition of IL‐1 b and TNF‐a production and decreases MPO activity in the site of inflammation.

Objective (s): Familial Mediterranean fever (FMF), an inherited autosomal recessive disorder, is frequently present among individuals of Mediterranean origin. Differences in the clinical manifestations of FMF between different ethnic groups have been documented. The aim of the present study was to determine the most common characteristics of FMF and the relationship between clinical findings and the most common mutant alleles of the MEFV gene in an Iranian Azeri Turk population.Materials and Methods: We analyzed clinical and genetic data from 415 patients identified as having FMF clinical symptoms and who were referred to the Molecular Genetics Laboratory of Tabriz/Iran over the last 3 years. The mutation type and clinical characteristics were determined for each patient.Results: The following primary clinical characteristics of the patients were observed: peritonitis was observed in 378 (93.8%), high‐grade fever in 351 (86.88%), arthritis in 215 (54.57%), pleuritis in 207 (53.49%), myalgia in 153 (41.69%), AA amyloidosis in 149 (40.16%), and erysipelas‐like erythema in 54 (14.96%) subjects. A positive response to colchicines treatment was noted in 374 (95.1%) patients including 303 patients with two mutated alleles and 71 patients with one identified mutation.Conclusion: In contrast to previous studies, there was no significant association between M694V mutation and development of amyloidosis. The M680I/M680I, M680I, M694I, and M694V/R761H genotypes were found to be associated with the development of amyloidosis. These results indicate that physicians need to pay careful attention to patients with asymptomatic or mildly symptomatic FMF with these genotypes.

Objective (s): To determine the effect of acetylcholine (ACh), pilocarpine, and atropine on pain evoked responses of pain excited neurons (PEN) and pain inhibited neurons (PIN) in hippocampal CA3 region of morphine addicted rats.Materials and Methods: Female Wistar rats, weighing between 230‐260 g were used in this study.Morphine addicted rats were generated by subcutaneous injection of increasing concentrations of morphine hydrochloride for six days. Trains of electrical impulses applied to the sciatic nerve were used as noxious stimulation and the evoked electrical activities of PEN or PIN in hippocampal CA3 area were recorded using extracellular electrophysiological recording techniques in hippocampal slices.The effect of acetylcholine receptor stimulation by ACh, the muscarinic agonist pilocarpine, and the muscarinic antagonist atropine on the pain evoked responses of pain related electrical activities was analyzed in hippocampal CA3 area of morphine addicted rats.Results: Intra‐CA3 microinjection of ACh (2 mg/1 ml) or pilocarpine (2 mg/1 ml) decreased the discharge frequency and prolonged the firing latency of PEN, but increased the discharge frequency and shortened the firing inhibitory duration (ID) of PIN. The intra‐CA3 administration of atropine (0.5 mg/1ml) produced opposite effect. The peak activity of cholinergic modulators was 2 to 4 min later in morphine addicted rats compared to peak activity previously observed in normal rats.Conclusion: ACh dependent modulation of noxious stimulation exists in hippocampal CA3 area of morphine addicted rats. Morphine treatment may shift the sensitivity of pain related neurons towards a delayed response to muscarinergic neurotransmission in hippocampal CA3 region.

Objective (s): Some evidence showed that calcitriol has an important role in regulating growth and differentiation of mesenchymal stem cells (MSCs). However, the interaction between mesenchymal stem cells and macrophage is not clear yet. The current study was done to investigate thein vitro effects of calcitriol on the interactions between bone marrow‐derived MSCs and rat macrophages.Materials and Methods: MSCs were isolated from rat bone marrow and pulsed with different concentrations of calcitriol (50, 100 and 200 nanomolar) for 24, 48 and 72 hr. Then, mesenchymal stem cells were co‐cultured with macrophages for 4 hr. Finally, macrophages were evaluated for ability to uptake neutral red, phagocytosis activity against opsonized yeast, respiratory burst and viability.Results: Our data showed that bone marrow‐derived MSCs pulsed with calcitriol may cause a significant increase in uptake of neutral red and phagocytic activity of opsonized heat killed baker' s yeast. Moreover, treatment of MSCs with calcitriol enhanced macrophage viability. Nevertheless, the respiratory burst of macrophages was significantly reduced in macrophages co‐cultured with calcitriol‐treated MSCs compared to control group.Conclusion: Calcitriol may accelerate and potentiate anti‐inflammatory M2 macrophage polarization by MSCs.

Objective (s): Cutaneous Leishmaniasis (CL) is a parasitic disease caused by various species of the flagellated protozoan, Leishmania. Regardless of the numerous studies, there are still serious challenges in the treatment of CL. This study aimed at evaluating the influence of a low dose ultraviolet B (UVB) radiation along with silver nanoparticles (AgNPs) on a mouse model of CL induced byLeishmania major.Materials and Methods: L. majorpromastigotes (MRHO/IR/75/ER) were extracted from infected mice spleens. Two months after subcutaneous injection of 2×106 promastigotes into the footpad of BALB/c mice, when the lesions were developed, the animals were divided into 4 groups including one control group and three study groups: AgNPs, UVB and UVB plus AgNPs. Spleen parasite burden was assessed on day 40 after the first treatment. The data were analyzed by Instat, Elida and SPSS 16 software programs.Results: The results showed the highest pronounced inhibitory effect in the group receiving AgNPs plus UVB. In addition, a significant difference was obtained between the group receiving AgNPs alone and the one with combinational therapy. The findings on parasite burden showed a significant difference between the control group and other treatment groups.Conclusion: It could be suggested that UVB in the presence of AgNPs, by inhibiting the spread of CL lesions and reducing the rate of visceral progression of the disease, provides a serious antileishmanial effect.

Objective (s): Eukaryotic translation initiation factor 4E (eIF4E) is overexpressed in cervical cancer (CC). However, the molecular mechanisms are unclear. This study aimed to investigate the molecular mechanism of eIF4E gene overexpression in CC.Materials and Methods: The human papillomavirus (HPV) type 18 E7 and eIF4E mRNAs were measured following knock down or overexpression of E7 gene by RT‐PCR and real‐time PCR. Cell counting kit‐8 assay was used to determine the cell proliferation. Flow cytometry was used to analyze the cell cycle and apoptosis. Transwell system was employed to determine the cell migration.Results: Overexpression of E7 gene increased eIF4E mRNA level by 24.3% (P<0.01) in HPV negativeC33A cells. Knock down of E7 decreased markedly eIF4E mRNA by 73% (P<0.01) in HPV18 positiveHeLa cells. Under the state of high expression of E 7, 1) up‐regulation of eIF4E drastically promoted the cell proliferation, cell cycle and cell migration, and inhibited the cell apoptosis.2) down‐regulation of eIF4E significantly inhibited the cell proliferation, cell cycle and the ability of cell migration, and also promoted the apoptosis of cervical cancer cells.Conclusion: HPV E7 induced eIF4E gene over transcription which might be a new marker for CC. The finding broadens the understanding of the CC carcinogenesis.

Objective (s): The current study was aimed to determine the bioactive constituents and biological effects of the Crataegus monogyna ethanolic extracts from bark, leaves and berries on hypercholesterolemia.Materials and Methods: Oleanolic acid, ursolic acid, quercetin and lupeol concentrations were quantified by HPLC. Total phenol content and radical scavenging activity of extracts were also measured. The hypocholesterolemic, antioxidant, and hepatoprotective effects of the extracts were examined in hypercholesterolemic rats and compared with orlistat.Results: The highest phenol content, oleanolic acid, quercetin and lupeol levels and free radical scavenging potency were found in the bark extract, and the highest ursolic acid level was found in the berries extract. Orlistat and extracts significantly (P<0.05) lowered the hypercholesterolemiaincreased serum level of hepatic enzymes and lipid peroxidation level. Hawthorn’s extracts protected from hepatic thiol depletion and improved the lipid profile and hepatic damages.Conclusion: Data suggested that hawthorn’s extracts are able to protect from hypercholesterolemiainduced oxidative stress and hepatic injuries. Moreover, the hypocholesterolemic effect of extracts was found comparable to orlistat.

Objective (s): miR‐125b has been identified as a tumor suppressor in many tumors, but its role in giant cell tumor (GCT) of bone remains poorly understood. The current study aimed to investigate the potential role and mechanism of miR‐125b in GCT.Materials and Methods: Expression levels of miR‐125b in GCT tissues were determined using RT‐PCR.The cell proliferation was surveyed by direct cell counting, MTS and CCK‐8, and the apoptotic cells were evaluated by Annexin V‐FITC and propidium iodine staining assay. The target gene expression was determined using RT‐PCR and western blot. Parathyroid hormone 1 receptor (PTH1R) 3’‐UTR was cloned into luciferase reporter plasmid to confirm direct targeting.Results: We found that miR‐125b was significantly down‐regulated in GCT tissues. Using both gainand loss‐of‐function analyses, we further revealed that miR‐125b suppressed GCT stromal cell proliferation and induced cell apoptosis. Furthermore, we revealed that PTH/PTHrP type 1 receptor is a direct and functional target of miR‐125b.Conclusion: Our results suggest that miR‐125b acts as a tumor suppressor through suppression of the PTH1R/RANKL signaling pathway. These findings contribute to our understanding of the functions of miR‐125b in GCT.

Objective (s): A Box‐Behnken design was used for evaluation of Eudragit coated diclofenac pellets. The purpose of this work was to optimize diclofenac pellets to improve the physicochemical properties using experimental design.Materials and Methods: Diclofenac was loaded onto the non‐pareil beads using conventional coating pan. Film coating of pellets was done at the same pan. The effect of plasticizer level, curing temperature and curing time was determined on the release of diclofenac from pellets coated with polymethacrylates.Results: Increasing the plasticizer in the coating formula led to decrease in drug release and increasing the curing temperature and time resulted in higher drug release. The optimization process generated an optimum of 35% drug release at 3 hr. The level of plasticizer concentration, curing temperature and time were 20% w/w, 55°C and 24 hr, respectively.Conclusion: This study showed that by controllinig the physical variables optimum drug release were obtained.

Objective (s): Based on the previous reports, silymarin can suppress nitric oxide, prostaglandin E2 (PGE2), leukotrienes, cytokines production, and neutrophils infiltration. Regarding the fact that inflammation plays an important role in neuropathic and formalin‐induced pain, it was assumed that silymarin could reduce pain. The present study investigates the analgesic effects of silymarin in chemical nociception and a model of neuropathic pain.Materials and Methods: Chemical nociception was produced by injection of 20 μl of formalin (0.5% formaldehyde in saline) into the plantar region of the right hind paw. A sciatic‐nerve ligated mouse was applied as the model of neuropathic pain and the antinociceptive response of silymarin was examined 14 days after unilateral nerve‐ligation using the hot plate test.Results: The intraperitoneal administration of silymarin (25, 50, and, 100 mg/kg) 2 hr prior to the intraplantar formalin injection suppressed the nociceptive response during the late phase of the formalin test significantly, but it was not in a dose‐dependent manner. Different doses of silymarin 14 days after unilateral sciatic nerve ligation in hot plate test did not induce obvious antinociception.Conclusion: Results of the present study indicated that repeated administration of silymarin prevents the formalin‐induced nociceptive behavior. However, it is not effective in the treatment of sciatic neuropathic pain.