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Background: Azoospermia is a common cause of male infertility. The latest hope for more affordable and safer therapies comes from stem cell research. In this study, the effect of mesenchymal stem cells derived from Wharton's jelly on spermatogenesis in azoospermic mice with scrotal hyperthermia was investigated.
Methods: For the experimental study, 24 adult male mice were divided into four groups:1. control (Cont), 2. scrotal hyperthermia (Hyp), 3. scrotal hyperthermia + DMEM (10μl) (Hyp/DMEM), 4. scrotal hyperthermia + conditioned medium (10μl) (Hyp/CM). A temperature of 43 °C was applied for 20 minutes every other day for two weeks to induce hyperthermia. The mice in the experimental group received 10µl of hJWSC-CM intraperitoneally every other day for 35 days after hyperthermia. Animals were sacrificed after the experiments to perform additional molecular, biochemical and stereological analyses.

Results: The results showed that CM significantly increased both the total amount of sperm and the number of testicular cells, including spermatids, primary spermatocytes, spermatogonia, Leydig cells, and Sertoli cells, compared with Hyp and Hyp/DMEM; in addition, the biochemical characteristics of testicular tissue were significantly higher in the Hyp/CM group compared with the hyperthermia-induced groups. Further analysis revealed that when the Hyp/CM group was compared with the Hyp and Hyp/DMEM groups, Sox9 protein expression and proliferative gene expression increased significantly and the percentage of apoptotic testicular cells and pro-inflammatory gene expression decreased significantly. 

Conclusion: These results suggest that realistic therapeutic approaches for reproductive and regenerative medicine could benefit from hWJSC-CM.

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Background: One of the most common cancers of the oral cavity is oral squamous cell carcinoma (OSCC). Previous studies have provided evidence that apolipoprotein C-I (Apo C-I) has oncogenic role beyond its classical function. Although it has been investigated in prostate, breast, and lung cancer, but there is no information available regarding Apo C-I expression in tumor and pri-tumor tissues of OSCC patients. Thus, the goal of present study was to unravel the expression of Apo C-I in OSCC and investigate its correlation with survival and grade of OSCC patients.
Methods: The Apo C-I mRNA level was investigated in tumor and pri-tumor tissues of 16 OSCC patients. In addition, the 34 paraffin-embedded tissues of OSCC patients and IHC technique were used to analyse the association of Apo C-I protein with survival of OSCC patients.
Results: The mRNA (P=0.0386) gene expression of Apo C-I showed statistically significant difference between tumor and pri-tumor tissues of OSCC patients. It seems that a high protein level of Apo C-I is related with poor survival of OSCC patients (P=0.04). The Apo C-I protein was positively correlated with tumor site (Pearson r= 0.485, P=0.0036) but not the grade (Pearson r= 0.2295, P=0.1917). Our data showed that high Apo C-I protein level might be correlated with poor survival of OSCC patients.
Conclusion: Our findings suggest that patients with high protein level of Apo C-I may have lower survival, making it as a potential prognostic factor for OSCC. However, further investigation is necessary to establish this concept.
 

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Introduction: To evaluate the cAMP -mediated IBMX (3-IsoButyle -1-Methyl Xanthin) and db-cAMP (dibutyryl cAMP) effects on differentiation of human Embryonic Stem Cells (hESCs) into nerve cells were the objectives of this study. Methods: We have used Royan H1 hESC- derived embryoid bodies with four treatment groups: six days treatment with IBMX (5×10 -4M) and db-cAMP (10 -9M) (referred to as cAMP), retinoic acid (RA, 10 -6M), IBMX + db-cAMP + RA, and control (no treatment). Immunocytochemistry was carried out for neural specific antibodies including β-Tubulin III, Microtubule Associated Protein 2 (MAP-2), Neurofilament Protein-Heavy chain (NF-H), Glial Fibrilary Acidic Protein (GFAP) and Synaptophysin as well as morphological studies. Semiquantitative RT-PCR was also used to evaluate gene expression involved in neurogenesis. Results: In the 4+6+4 days the neuronal process were apparently observed. Immunocytochemical studies using nerve specific antibodies for proteins such as β- Tubulin III, MAP-2, NF-H, GFAP and Synaptophysin showed the presence of these neuronal and astrocyte markers in differentiated cells by cAMP. Evaluation of expression of genes involved in neurogenesis showed that Hash1, Synaptophysin, β-Adrenergic Receptor and Acetylcholine Receptor- which were silent in embryoid bodies - switched on after treatment with cAMP and/or RA. Relative expression of nerve specific genes showed a significant enhancement in expression of Synaptophysin, NFM and β-Adrenergic Receptor during differentiation, which, with the enhancement in cAMP treated groups were more than those treated with RA and control (p < 0.05). Conclusion: In conclusion, this study showed that cAMP could be a neurogenic agent for human embryonic stem cells differentiation.

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Introduction: Since organophosphorus compounds (OP) are toxic and designed to destroy insects and pest species, there are many hazards associated with their use. Although, the main target site of these compounds is acetylcholinesterase (AChE), however it has become increasingly evident that OPs have also other direct effects on cellular processes. In the present study, the effects of low concentrations of paraoxon and its interaction with forskolin, an activator of protein kinase A (PKA), were studied on Ca2+ spike configuration and frequency in neurons of snail Caucasotachea atrolabiata. Methods: Subesophageal ganglia neurons were recorded in current clamp mode in Na+ free Ringer solution that contained voltage dependent potassium channel blockers, 4AP and TEA. Results: Paraoxon (0.3-0.6 μM) decreased the duration of spontaneous Ca2+ spikes. This effect was seen with a suppression of single spike AHPs, leading to an increment in firing rate. Paraoxon induced hyperactivity appeared to be a consequence of decrease in Ca2+ influx during spikes which is the main determinant of AHP duration by activating Ca2+ dependent potassium channels. Forskolin (25 μM), in the absence of a significant change in spike duration, decreased the duration of single spike AHPs and increased the frequency of spikes. After forskolin application, paraoxon decreased the duration of Ca2+ spikes and AHPs, and increased the activity. However, these effects, especially on spike duration, were not as pronounced as in the absence of forskolin. Conclusion: These findings suggest that although forskolin, similar to paraoxon, decreases the AHP and increases the frequency of spikes but it employs mechanism(s) different from paraoxon which also oppose the effects of paraoxon on Ca2+ spikes configuration and frequency.

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Introduction Numerous studies have demonstrated the presence of potassium selective channels in membranes internal organelles. These channels are essential to a large variety of cellular processes including intracellular 2+ a signaling, protein recycling, charge neutralization and cell protection. In contrast to the sarcoplasmic reticulum + here potassium channels have been clearly identified, there is little evidence for K selective cannels in RER hepatocytes. The aim of this study is to find an evidence for presence of potassium channel in endoplasmic ticulum and considering the pharmacological and biophysical properties of this channel. Methods: Hepatocytes RER vesicles were isolated by homogenizing rat liver followed by several centrifuging eps and then incorporated into the bilayer lipid membrane (BLM). The BLM was formed by painting osphatidylcholine across the 350 ?m aperture seperating two chambers (cis chamber containing 200 mM KCl + d trans chamber containing 50 mM KCl). Single channel recordings were used to indicate the presence of K annels. Results: Single channel recordings revealed the existence of a cation selective channel with high permeability + K and 599 pS conductance. The current–voltage relation was linear. The open probability was strongly voltage pendent, showing higher values at positive voltages and lower at negative voltages. A subconductance state out 60 % of fully open state was observed at all voltages. The cationic channel showed an inhibition by 4- + innopyridine (non specific K channel blocker) + Conclusions: In this study we have established an evidence for existence of large conductance K channel in e hepatocyte ER vesicles.

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Introduction: (S)- 3,5-Dihydroxyphenylglycine (DHPG) is an agonist for group I metabotropic glutamate receptors. DHPG-induced synaptic depression of excitatory synapses on hippocampal pyramidal neurons is well known model for synaptic plasticity studies. The aim of the present study was to examine the effects of DHPG superfusion on excitatory synapses on pyramidal and fast-spiking GABAergic cells (FS-GABA) of layer II/III of mice visual cortex. Methods: Effects of DHPG was examined in visual cortical slices of GAD67-GFP knock-in mice using whole-cell recordings of excitatory postsynaptic potentials (EPSPs) in layer II/III cells evoked by layer IV stimulation. In part of experiments, long term potentiation (LTP) was induced by theta burst stimulation (TBS) paired with postsynaptic depolarization. Results: DHPG induced potentiation of EPSPs of FS-GABA neurons in dose- and use-dependent manners but it has no effect on pyramidal cell excitatory synapses. An antagonist for type 5 metabotropic glutamate receptors (mGluR5) blocked DHPG-induced LTP, while an antagonist for mGluR1 was not effective. This potentiation and TBS-induced LTP occluded each other. Conclusion: Based on important role of FS-GABA cells in cortical neuronal circuit, mGlur5-dependent LTP may play a role in, enhancement or maintenance of synchronized activity of cortical pyramidal neurons.

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Introduction: Nicotine is the psychoactive substance responsible for establishing and maintaining smoking dependence. CYP2A6 is the primary enzyme that inactivates nicotine to cotinine .Genetic variation in CYP2A6 accounts for some of the inter-individual variability in nicotine metabolism and has been indicated to influence smoking behavior and dependence. Therefore, the aim of this study was to examine whether there is a relationship between CYP2A6 genetic polymorphism and smoking dependence in an Iranian population. Methods: We assessed 118 male non-smokers (1-99 cigarette/ lifetime) and 133 dependent current smokers for demographic, cigarette use history and DSM-IV dependence. CYP2A6 alleles associated with decreased nicotine metabolism (* 2, *4, or *9 allele) were determined using allele-specific nested PCR. Genotypes were grouped into slow metabolizers (one or two copies of *2 or *4, or two *9 alleles), intermediate (one *9 allele), and normal (have no copies of *2 , *4 , or *9 alleles). Results: Intermediate nicotine metabolizers were at higher risk for becoming a dependent smoker odd ratio (OR = 3.71 p=0.009). Slow metabolizers had a significantly lower age of first smoking compared to normal and intermediate metabolizers (p = 0.037). Cigarette consumption and the degree of smoking dependence were not significantly different among smokers with different CYP2A6 genotypes. Conclusion: In Iranian population, the risk for becoming a dependent smokers increases with genotypes for intermediate metabolism of nicotine and slow nicotine metabolizers experience smoking in lower ages. These findings increase our understanding of the effect of CYP2A6 genotypes on smoking dependence in Iranian population and may help us to develop new strategies for quitting smoking.

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Abstract The S362A mutation block ROMK2 (Kir1.1b) endocytosis in Xenopus laevis oocyte membrane . Saeed Hajihashemi1 , 1-Assistant professor, PhD in Physiology, Department of Physiology, School of Medical science, Arak University of Medical Sciences. Introduction: ROMK channel is localized on the apical membrane of the nephron. Recent studies suggest that endocytosis of ROMK channels is important for regulation of K+ secretion in cortical collecting ducts. In this study the effect of S362A mutation is examined on the membrane turnover and stability of ROMK2 channel when expressing in Xenopus laevis oocytes. Materials and Methods: In this experimental study oocytes were isolated by standard protocols using collagenase (Type 1A). Mutations of the cytoplasmic termini of ROMK2 were constructed using the quik-change approach for site-directed mutagensis. Xenopus oocytes were injected with cRNA encoding ROMK2 or S362A mutant three days prior to treatment with BFA solution (time 0). Brefeldin A (BFA) was added to the OR3 medium (+BFA) at concentrations of 25µM (inhibit insertion of new proteins into the cell membrane) or ethanol as BFA vehicle (-BFA). Two-electrode voltage clamp (TEVC) was used to measure oocyte ROMK-dependent currents and membrane potential. Data was analyzed using Student’s t-tests or ANOVA as appropriate. Results: Incubation of oocytes expressing ROMK2 channels in 25 μM BFA caused a reduction in the currents and membrane voltage. In oocytes expressing the S362A mutant, there was no decay in current and membrane voltage after 48 hours incubation with BFA at 25 µM. The fractional current for ROMK2 at 48h following treatment of oocytes with BFA was 0.24 0.05 (n=24) which was significantly different to S362A mutant (0.96 0.05 n=24). Conclusion: These results show that the S362A mutation increases the general stability of ROMK and renderes the protein resistant to endocytosis, consistent with the idea that there is an interaction between the C-terminal of ROMK2 and components of the endocytotic pathway. A functional PDZ domain (the S-E-V) plays a key role in determining stability of ROMK. Key words: ROMK2, S362A mutant, BFA, PDZ domain

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Introduction: Previous investigations have indicated that the presence of BIO (6-Bromoindirubin-3-Oxime) in medium of some cell culture enhances the cell proliferation and viability. The aim of the present study was to investigate the BIO effects on in vitro expansion of rat marrow-derived mesenchymal stem cells (MSCs) culture. Methods: In the present experimental study, bone marrow cells from 7 rats were plated in the presence of 0.05, 0.01, 0.1, 1 and 1.5 µM of BIO and expanded through three successive subcultures. During the cultivation period, the cultures were statistically compared in terms of some indices of cell growth including the diameter and number of colonies, population doubling number (PDN) and the number of viable cells. Passaged-3 cells from all groups were examined whether or not they could differentiate into bone and adipose cells. Results: According to our results, the largest colonies were formed in the cultures with 0.1 and 1 µm BIO with diameter of respectively 1262.27±43.96 and 1335.71 ± 19.16 micrometer (P<0.05). These two groups were also superior in terms of the colonies numbers. During three successive passages, significantly more PDN was occurred in 0.1 and 1 µM BIO- treated cultures than the others (P<0.05). Additionally in these two BIO-treated cultures, the number of viable cells was significantly higher compared to other BIO-treated cultures as well as the control group (P<0.05). Alizarin red staining for bone and oil red for adipose cells indicated the differentiation potential of the cells in all studied groups Conclusion: Taken together it seems that the BIO presence in marrow cell cultures enhances the cell in vitro expansion and viability.

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Introduction: The Serum/Glucose deprivation -induced cell death in cultured PC12 cells represents a useful in vitro model for the study of brain ischemia and neurodegenerative disorders. Nigella sativa L. has been known as a source of antioxidants. To elucidate the neuroprotective actions of N. sativa extract in vitro, we studied the effect of N. sativa extract on cultured PC12 cells under serum/glucose deprivation conditions. Methods: PC12 cells were cultured in DMEM medium containing 10% (v/v) fetal bovine serum, 100 units/ml penicillin, and 100 μg/ml streptomycin. Cells were seeded overnight and then deprived of serum/glucose for 6 and 18 h. Cells were pretreated with different concentrations of N. sativa extract (7.81-250 μg/ml). Cell viability was quantitated by MTT assay. Intracellular ROS production was measured by flow cytometry using 2', 7’-Dichlorofluorescin diacetate (DCF-DA). Results: Depriving the PC-12 cells of serum/glucose caused prominent cell toxicity at least after 6 and 18 h. Pretreatment of PC12 cells with N. sativa (7.81-250 μg/ml) could reduce serum/glucose deprivation-induced cytotoicity in PC12 cells after 18 h. The experimental results suggest that N. sativa extract protects the PC12 cells against Serum/Glucose deprivation-induced cytotoxicity. Conclusion: Our findings might raise a possibility of potential therapeutic application of N. sativa extract for preventing and treating cerebral ischemic and neurodegenerative diseases.

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Introduction: S100B is a protein released from astrocytes, which controls neuroglial relationship and probably plays a role in memory and synaptic plasticity. Study of the functional relationship between astrocytes and neurons is of great importance. The present study was conducted to evaluate the effect of S100B infusion into the CA1 region of the hippocampus, on memory performance in male rats. Methods: In this study, 40 male wistar rats were used. Animals were bilaterally implanted with indwelling cannulae in the CA1 region of the hippocampus. Seven days after surgery, animals were trained in a step-down passive avoidance task (0.5 mA, 100 Hz, 5 sec). Immediately after the training, animals received 0.5 μl infusion of saline or S100B (5, 50, 500, or 5000 ng) bilaterally. Twenty four hours later step-down first latency and total time spent on platform were measured as learning and memory indices. Results: The infusion of 5 ng S100B induced a significant increase in step-down first latency (p< 0.01), and also increased the total time spent on the platform compared to the control group (p < 0.001). Surprisingly, animals which received doses of 500 and 5000 ng showed a significant decrease in both indices compared to the control group (p< 0.001). Conclusion: Our findings indicate that astrocytic S100B protein has modulatory effects on memory, in a way that in nanogram doses facilitates, but in micrograms impairs memory in passive avoidance task.

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Introduction: Thrombosis and the blockage of blood vessels with clots, can lead to acute myocardial infarction and some times even death. Aside from surgical interventions to remove the blockage, the only available treatment is the administration of thrombolytic agents to dissolve the blood clot. Streptokinase (SK) is the most commonly used fibrinolytic drug for this purpose. However, SK has some disadvantages including immunogenicity, short half-life and hemorrhage induction. PEGylation of pharmaceutical proteins by the incorporation of cysteine amino acid is a novel method to decrease their immunogenicity and also to increase their stability and half-life. The ultimate goal of this study was designing and construction of the cysteine analogues of full-length and truncated forms of SK, which posses less hemorrhagic side effects due to fibrin specificity. Methods: By application of PCR-based site-directed mutagenesis technique, mutants of SK genes, harboring the transversion of AGC codon (serine) to TGC (cysteine), which encoded full-length (amino acids 1-414) and truncated (amino acids 60-386 and 143-386) proteins were established and cloned in pET41a plasmid. Expression of the recombinant SKs was achieved through the induction of E. coli transformants. Produced proteins were confirmed by western blotting, purified by affinity chromotography and finally evaluated for their biological activity. Results: Mutant SK genes were efficiently expressed and due to the fusion of vector-derived His-tag the recombinant full-length and truncated proteins were easily purified. Also despite the replacement of serine to cysteine in the position of 208, the biological activity of the new recombinant protein was still maintained. Conclusion: The produced mutants of this study provide the possibility of establishing the cysteine specific PEGylation process and improvement of clinical activity of streptokinase protein.

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Introduction: Recent studies have shown the presence of Cl- channels in heart and liver mitochondrial membranes. In this work, we have characterized the functional profile of a Cl- channel from rat brain mitochondria. Methods: After removing and homogenizing the rat brain, the supernatant was separately centrifuged in MSEdigitonin, H2O and Na2CO3 and mitochondrial inner membrane vesicles were obtained in MSE solution. L-α- Phosphatidylcholine (membrane lipid) was extracted from fresh egg yolk. Bilayer lipid membranes were formed in a 150 μm diameter hole. All recordings were filtered at 1 kHz and stored at a sampling rate of 10 kHz for offline analysis by PClamp10. Statistical analysis was performed based on Markov noise free single channel analysis. Results: Brain mitochondrial inner membrane preparations were subjected to SDS-PAGE analysis and channel protein reconstitution into planar lipid bilayers. Western blotting and antibodies directed against various cellular proteins revealed that the mitochondrial inner membrane fractions did not contain specific proteins of the other subcellular compartments except a very small fraction of endoplasmic reticulum. Channel incorporation into the planar lipid bilayers revealed an anion selective channel with a conductance of 301 pS in 200 mM KCl cis/50 mM KCl trans. The channel open probability appeared to be voltage dependent and the channel was active between the voltages of -40 and +20 mV. Adding 10 μM DIDS to the side corresponding to the cell internal medium caused a strong inhibition of the channel activity. Conclusion: This channel is likely to be involved in maintaining proper pH, membrane potential, ATP synthesis, and cell protection.

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Introduction: we have recently reported the presence of two potassium currents with 598 and 368 pS conductance in the rough endoplasmic reticulum (RER) membrane. The 598 pS channel was voltage dependent and ATP sensitive. However, the 368 pS channel was rarely observed and its identity remained obscure. Since cationic channels in intracellular organelles such as mitochondria and RER play important roles in intracellular signaling and cellular protection, studying their biophysical and pharmacological properties is important. Methods: we used channel incorporation into bilayer lipid membrane method. L-α-Phosphatidylcholine (PC), a membrane lipid, was extracted from fresh egg yolk. Bilayer lipid membrane was formed in a 150 μm diameter hole. Rough endoplasmic reticulum vesicles were obtained from liver after homogenization and several centrifugations. All recordings were filtered at 1 kHz and stored at a sampling rate of 10 kHz for offline analysis by PClamp9. Statistical analysis was performed based on Markov noise free single channel analysis. Results: A 364 pS potassium channel was identified which was voltage independent in +50 to -50 mV voltages. In all voltages, open probability (Po) was over 0.9. Nonspecific inhibitor of K+ channels, 4-aminopyridine (4-AP, 20 mM), inhibited the channel activity. However, intracellular nucleotide like ATP had no effect on channel gating. Conclusion: We observed a new potassium channel with 346 pS conductance in RER membrane that unlike the 598 pS K channel was not ATP sensitive. This channel may play an important role in Ca2+ homeostasis and cell protection.

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Introduction: Anethole is the main constituent of Pimpinella anisum L. (anise), a herbaceous annual plant which has several therapeutic effects. In the folk medicine, anise is employed as an antiepileptic drug. Specifically, this study was focused on the cellular effect of anethole, an aromatic compound in essential oils from anise and camphor. Anethole has various physiological effects on the cardiovascular system and smooth and skeletal muscles. However, despite these persistent effects, there is little information available about the actions of anethole on nerve cells. Therefore, a major goal of the present research was to investigate the possible cellular mechanisms underlying the effect of anethole on the neural excitability and action potential characteristics in snail neurons. Methods: Intracellular recordings were made under the current clamp condition on F1 cells of Helix aspersa. Following extracellular application of anethole (0.5% or 2%), changes in the firing pattern and action potential parameters were assessed and compared to control condition. Results: Application of anethole (0.5% and 2%) led to a significant increase in the action potential amplitude and a reduction in the peak area and time to peak. In the presence of 0.5% anethole, the after hyperpolarization (AHP) amplitude was significantly decreased, while the firing frequency of neurons was increased. However, 2% anethole did not affect the AHP amplitude, but significantly reduced the firing frequency of action potentials. Conclusion: Based on the effect of anethole on the action potential parameters, it can be concluded that it probably affects the voltage gated ion channels function, including Ca2+ channels and/or Ca2+ dependent K+ channels activity.

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Introduction: Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease. In the present study, we investigated the response of subventricular zone (SVZ) adult stem cells in the experimental autoimmune encephalomyelitis (EAE), an animal model of MS, and also the differentiation fate of these stem cells. Methods: Mice were immunized with MOG peptide emulsified in complete Freund's adjuvant (CFA) and pertussis toxin (PT). Control mice received CFA and PT. To study SVZ stem cells migration, mice received seven i.p. injections of BrdU at 2-h intervals on the day before EAE induction. Demyelination was studied using specific staining with luxol fast blue. The number of BrdU+ cells in SVZ and olfactory bulb (OB) was counted using immunohistochemical staining. To understand the fate of the stem cells, NG2 marker was used to detect oligodendrocyte precursors. Results: Lumbar spinal cord of EAE animals showed significant demyelination and the volume of demyelinated areas was increased on days 14 to 21 post lesion. In the EAE group, more Brdu+ cells were observed in the OB compared to the SVZ. The number of Brdu+/NG2 + cells in OB was significantly increased after EAE induction. Conclusion: The demyelinating context of EAE promotes the migration of SVZ stem cells to the OB. These cells mostly differentiate to oligodendrocyte precursors and may contribute to myelin repair.

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Introduction: Cyclophosphamide (CP) is used for the treatment of various cancers. In spite of its therapeutic importance, a wide range of adverse effects such as reproductive toxicity has been observed following the administration of this drug in human and experimental animals. In the current study, we have investigated the adverse effects of CP on morphology and histology of testis rats. Methods: Twenty-one male Wistar rats were selected and randomly divided into 3 groups. CP was used at a dose of 6.1 mg/kg/day, (i.p.) for 50 days. At the end of the treatment, the histological and biochemical changes in testis, as well as sperm count and motility were assessed. Results: Testicular weight, sperm count and motility as well as serum testosterone concentration were significantly decreased whereas malondialdehyde (MDA) level was significantly increased in CP group compared with those in the control and sham groups. In addition, histological studies of testis structure showed that seminiferous tubules of testis were severely damaged in the CP group. CP increased the number of sloughing tubules and interstitial space, while it decreased seminiferous tubular diameter (STD), seminiferous epithelial height (SE), tubule differentiation index (TDI) and spermiation index (SPI). Conclusion: The results suggest that cyclophosphamide affect fertility parameters and cause testis atrophy after chronic treatment.

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Introduction: In this study, the effect of para-nonylphenol as an environmental pollutant on viability, morphology and proliferation of bone marrow mesenchymal stem cells was investigated. Methods: Bone marrow mesenchymal stem cells of rat were treated with the 0.5, 1, 2.5, 3.5 and 5 μM of paranonylphenol for a period of 21 days, then the viability of the cells were estimated using trypan blue and MTT methods. After choosing the effective dose, the integration of the DNA was investigated using comet assay and agarose gel electrophoresis. Mechanisms of cell death were also investigated by TUNEL assay and presence of caspase activity. Results: The results showed that para-nonylphenol caused significant dose dependent reduction of viability and proliferation of the cells. Comet assay, agarose gel electrophoresis and TUNEL test showed that the DNA of the cells were damaged and broken after treatment with 0.5 and 2.5 μM of para-nonylphenol. In addition, activated caspase-3 was observed in the cytoplasm of treated cells. Conclusion: This study showed that a very low concentration of para-nonylphenol has drastic effects on bone marrow mesenchymal stem cells. This chemical is used in formulation of cosmetics and detergents and therfore may have detrimental effects on the viability and proliferation of stem cells.

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It is well known that the metabolic factors play an important role in the regulation of angiogenesis. Increased metabolic activity leads to decreased oxygen levels and causes tissue hypoxia. Hypoxia starts different signals to stimulate angiogenesis and promotes oxygen delivery to tissues. It has been suggested that released adenosine from hypoxic tissues plays a vital role in angiogenesis. In this context, the following feedback control hypothesis is proposed: Under hypoxic conditions, membrane 5´-nucleotidase dephosphorylates AMP to adenosine in the extracellular space of a cardiomyocyte, hepatocyte or other parenchymal cells. Extracellular adenosine binds and stimulates adenosine receptor that leads to the release of vascular endothelial growth factor (VEGF) from the parenchymal cell. Binding of VEGF to its receptor on the surface of endothelial cells activates proliferation and migration of these cells. Adenosine can also activate the proliferation of vascular endothelial cells through mediating other anti and proangiogenic growth factors. Adenosine can also induce vasodilation and play a role in the vascular growth and remodeling. After establishment of a new capillary network, adenosine, VEGF and other pro and antiangiogenic growth factors return to near basal concentrations, and then the angiogenesis process terminates. In some conditions up to 50–70% of the hypoxia-induced angiogenesis mediates through adenosine. Hence, the main aim of this review is to focus on the physiological role of adenosine and its receptors in the induction of angiogenesis under hypoxic conditions

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Introduction: Physalis alkekengi (Solanaceae) is a rich source of various antioxidants. There are some reports that show P. alkekengi has been used for treatment of a wide range of diseases including gout and inflammation. Xanthine oxidase plays a crucial role in gout. Many natural compounds such as various flavonoids have been reported to have inhibitory effect on xanthine oxidase. Methods: Different parts of P. alkekengi including leave, calyx, green and orange fruits at different stages of plant growth were gathered from around the Tonekabon, Iran. Then, they were dried in the dark and powdered. Inhibitory effect of various plant extracts on xanthine oxidase activity was measured. Soluble sugar and ascorbic acid contents of plant samples, and their correlation with xanthine oxidase inhibitory effect were also determined. Results: All extracts from different parts of P. alkekengi at the concentration of 0.3 mg/ml had inhibitory effect on xanthine oxidase activity with different degrees from 45% (leaves in the vegetative stage) up to 86.86% (leaves in the green fruit stage and calyxs). The leaves, fruits and calyx stage of maturity contained the highest amount of soluble sugar. Also, maximum amount of total ascorbic acid was displayed in an orange calyx, 12.65 mg.fw-1. Conclusion: These results suggest that extracts of P. alkekengi at different phonological stages have high inhibitory effects on xanthine oxidase activity and they are valuable sources of antioxidant compounds. The green fruit, green calyx and orange calyx had the highest inhibitory effects on the xanthine oxidase activity. Therefore, they are recommended as the most appropriate tissues for the next pharmacological studies.