Effects of Thymbra spicata extract and Thymol on morphine withdrawal syndrome in mice (insights to the liver function, antioxidant, and behavioral responses).

Safe chemicals for drug withdrawal can be extracted from natural sources. This study investigates the effects of clonidine and Thymbra spicata extract (TSE) on mice suffering from morphine withdrawal syndrome. Thymol, which is the active constituent in TSE, was also tested. A total of 90 mice were divided into nine groups. Group 1 was the control group, while Group 2 was given only morphine, and Group 3 received morphine and 0.2 mg/kg of clonidine. Groups 4-6 were given morphine along with 100, 200, and 300 mg/kg of TSE, respectively. Groups 7-9 received morphine plus 30, 60, and 90 mg/kg of Thymol, respectively, for 7 days. An oral naloxone challenge of 3 mg/kg was used to induce withdrawal syndrome in all groups. Improvement of liver enzyme levels (aspartate aminotransferase, alkaline phosphatase, and alanine transaminase) (p < .01) and behavioral responses (frequencies of jumping, frequencies of two-legged standing, Straub tail reaction) (p < .01) were significantly observed in the groups receiving TSE and Thymol (Groups 4-9) compared to Group 2. Additionally, antioxidant activity in these groups was improved compared to Group 2. Nitric oxide significantly decreased in Groups 4 and 6 compared to Groups 2 and 3 (p < .01). Superoxide dismutase increased dramatically in Groups 5, 8, and 9 compared to Groups 2 and 3 (p < .01). Groups 5-9 were significantly different from Group 2 in terms of malondialdehyde levels (p < .01). Certain doses of TSE and Thymol were found to alleviate the narcotics withdrawal symptoms. This similar effect to clonidine can pave the way for their administration in humans.

Thymol Nanopolymer Synthesis and Its Effects on Morphine Withdrawal Syndrome in Comparison With Clonidine in Rats.

The drug delivery system is valuable in the treatment of the disease. A nanopolymer as a thymol and Thymbra spicata release system was synthesized and its effects on morphine withdrawal syndrome in comparison with clonidine in rats were studied. The nanopolymer was characterized by different methods, namely, IR, HNMR, CNMR, GPC, DLS, and AFM. Thymol in T. spicata extract was assessed. The loading and release rate of thymol and T. spicata extract on the nanopolymer were evaluated by HPLC. The median lethal dose (LD50) of the T. spicata extract, thymol, extract nanopolymer, and thymol nanopolymer was studied. The frequency of jumping, rearing, and teeth chattering in naloxone-induced morphine withdrawal syndrome was studied. Synthesized nanopolymer was desirable as a carrier for the drug. The loaded amount of extract and thymol on nanopolymer was estimated 55 ± 3.2% and 48 ± 2.6% and the drug released was 71 and 68%, respectively. LD50 of the T. spicata extract, thymol, extract nanopolymer, and thymol nanopolymer was 975, 580, 1,250, and 650 mg/kg, respectively. This study showed that thymol nanopolymer was more effective than clonidine to reduce the frequency of morphine withdrawal symptoms. Our results suggest that T. spicata extract, thymol, extract nanopolymer, and thymol nanopolymer are mighty in reducing the narcotic withdrawal signs. The mechanism of action and therapeutic potential is maybe similar to clonidine.

The Effect of Prosopis farcta and Its Bioactive Luteolin on the Hippocampus of Mice after Induced Ischemia Reperfusion.

BACKGROUND: Ischemia plays an important role in increasing damage to the nervous system. This study aimed to evaluate the effect of Prosopis farcta (PFE) and its bioactive luteolin (Lu) and forced swimming exercise on the hippocampus of mice after induced ischemia reperfusion. METHODS: The bioactive component of PFE (Lu) was identified by HPLC. Fifty-six male mice were divided into different groups. Ischemia was induced by ligation of the common carotid artery. After mice training (swimming exercise, 8 weeks) and consuming PFE and Lu, the mice's memory ability was evaluated in the shuttle box. Histological examination was performed by Nissel staining and immunohistochemistry. RESULTS: Results showed that the ischemic mice exercised and treated with PFE and Lu had higher step-through latency (STL) compared with the nonexercised mice, and this was confirmed with time spent in the dark compartment (TDC). The number of dark cells in the ischemic group exercising and receiving PFE and Lu decreased compared to that of the other groups in the hippocampus. DCX protein expression was increased in nonexercised groups compared to that of the exercised groups and those treated with PFE and Lu, while NeuN decreased. CONCLUSIONS: Forced swimming exercise following ischemia, as well as consumption of PFE and Lu, has reduced cell death and increased neurogenesis in the hippocampus and thus may help improve memory in ischemia.

Transdifferentiation of Human Umbilical Cord-Derived Mesenchymal Stem Cells in Dopaminergic Neurons in a Three-Dimensional Culture.

Introduction: The induction of human umbilical cord-derived mesenchymal stem cells (HUC-MSCs) toward dopaminergic neurons is a major challenge in tissue engineering and experimental and clinical treatments of various neurodegenerative diseases, including Parkinson disease. This study aims to differentiate HUC-MSCs into dopaminergic neuron-like cells. Methods: Following the isolation and characterization of HUC-MSCs, they were transferred to Matrigel-coated plates and incubated with a cocktail of dopaminergic neuronal differentiation factors. The capacity of differentiation into dopaminergic neuron-like cells in 2-dimensional culture and on Matrigel was assessed by real-time polymerase chain reaction, immunocytochemistry, and high-performance liquid chromatography. Results: Our results showed that dopaminergic neuronal markers' transcript and protein levels were significantly increased on the Matrigel differentiated cells compared to 2D culture plates. Conclusion: Overall, the results of this study suggest that HUC-MSCs can successfully differentiate toward dopaminergic neuron-like cells on Matrigel, having great potential for the treatment of dopaminergic neuron-related diseases.

Melatonin ameliorates testes against forced treadmill exercise training on spermatogenesis in rats.

INTRODUCTION: It is well documented that some forced exercises can have bad effects on the genital system. Melatonin is a potent antioxidant that is effective in reducing the physical stress.

Potential role of human chorionic gonadotropin supplementation in spermatogenesis in rats subjected to forced swimming exercise.

AIM: The aim of this study was to evaluate the supportive effect of human chorionic gonadotropin (hCG) on the quality of spermatogenesis, including count, motility, morphology, viability and apoptosis of sperm following forced swimming exercise. MATERIALS AND METHODS: Twenty-four adult male Sprague-Dawley rats were used in this study. All rats were divided into four groups: control group; swimming exercise group (S); hCG administration group and swimming (SG) with hCG administration group (G). The experimental group was trained to force swimming stress for 10 min for 6 days. Then the sperm quality parameters were measured after dissection and epididymis removal. Spermatogenesis and germ cell apoptosis were evaluated by using Miller & Johnsen's score and TUNEL staining respectively. RESULTS: Results showed the count (control: 113±3.1, S: 74±1.9, G: 111±3, and SG: 103±2.4), motility (control:  93±2, S: 67±2.8,G: 90±2.7, and SG: 78±1), morphology (control: 89±3%, S: 47±2.4%, G: 90±3.1%, and SG: 67±1.1%), and viability of sperm (control: 91±2.9, S: 50±2, G: 91±1.9, and SG: 70±1.3) in swimming exercised-hCG administered group, significantly enhanced by hCG treatment compared to the swimming exercise group (p≤0.01). Also the number of apoptotic germ cells significantly decreased in swimming exercised-hCG administered group compared to the swimming exercised group. CONCLUSIONS: These results suggest that administration of hCG can protect the testes against the detrimental effect of forced swimming exercise in adult male rats.

Homing of Super Paramagnetic Iron Oxide Nanoparticles (SPIONs) Labeled Adipose-Derived Stem Cells by Magnetic Attraction in a Rat Model of Parkinson's Disease.

INTRODUCTION: Stem cell therapies for neurodegenerative diseases such as Parkinson's disease (PD) are intended to replace lost dopaminergic neurons. The basis of this treatment is to guide the migration of transplanted cells into the target tissue or injury site. The aim of this study is an evaluation of the homing of superparamagnetic iron oxide nanoparticles (SPIONs) labeled adipose-derived stem cells (ADSC) by an external magnetic field in a rat model of PD. METHODS: ADSCs were obtained from perinephric regions of male adult rats and cultured in a DMEM medium. ADSC markers were assessed by immunostaining with CD90, CD105, CD49d, and CD45. The SPION was coated using poly-L-lysine hydrobromide and transfection was determined in rat ADSC using the GFP reporter gene. For this in vivo study, rats with PD were divided into five groups: a positive control group, a control group with PD (lesion with 6-HD injection), and three treatment groups: the PD/ADSC group (PD transplant with ADSCs transfected by BrdU), PD/ADSC/SPION group (PD transplant with ADSCs labeled with SPION and transfected by GFP), and the PD/ADSC/SPION/EM group (PD transplant with ADSCs labeled with SPION and transfected by GFP induced with external magnet). RESULTS: ADSCs were immunoreactive to fat markers CD90 (90.73±1.7), CD105 (87.4±2.9) and CD49d (79.6±2.6), with negative immunostaining at the hematopoietic stem cell marker (CD45: 1.4±0.4). The efficiency of cells with SPION/PLL was about 96% of ADSC. The highest number of GFP-positive cells was in the ADSC/SPION/EM group (54.5±1.3), which was significantly different from that in ADSC/SPION group (30.83±3 and P<0.01). CONCLUSION: Transfection of ADSC by SPION/PLL is an appropriate protocol for cell therapy. External magnets can be used for the delivery and homing of transplanted stem cells in the target tissue.

Retinoic acid and fibroblast growth factor-2 play a key role on modulation of sex hormones and apoptosis in a mouse model of polycystic ovary syndrome induced by estradiol valerate.

OBJECTIVE: The main goal of the present study is to investigate the effects of retinoic acid and fibroblast growth factor-2 on serum levels of FSH and LH, histology, and apoptosis in the mouse model of Poly Cystic Ovary Syndrome (PCOS). MATERIALS AND METHODS: 80 female NMRI mice have been randomly divided into eight groups. Group 1 received normal saline as a control, and Group 2 received estradiol valerate (EV) at 4 mg/100 g of body weight. Moreover, Groups 3-4 were administered with RA (a dose of 0.05 µg/µl) and FGF2 (a dose of 0.01 µg/kg), respectively. Groups 5 and 6 respectively received the EV plus the RA (0.05 µg/µl) and FGF2 (0.01 µg/kg). Group 7 received the RA and FGF2 at doses corresponding to healthy mice, and Group 8 received the EV plus the RA + FGF2 (similar to previous doses). RA and FGF2 were injected three times per week for four weeks. Finally, histological and immunohistological parameters of the ovary were evaluated. RESULTS: The study revealed that both single and combined injection of fibroblast growth factor-2 (FGF2) and retinoic acid (RA) in groups 5, 6, and 8 significantly reduced follicular diameters compared to group 2. Measurements confirmed that simultaneous injection of RA and FGF2 into polycystic mice significantly increased antral follicles, corpus luteum (CL), epithelial thickness, and oocyte diameter as well as decreased cystic follicles. Positive TUNEL cells that were considerably increased in the antral follicle of group 2 significantly decreased in the RA and FGF2 recipient groups, either alone or in combination. Besides, the injection of FGF2 increased preantral follicles and CL. CONCLUSION: The findings of the present investigation reveal that injection of RA and FGF2 has both protective and ameliorative effects that can promise new therapies for women with PCOS.

PuraMatrix hydrogel enhances the expression of motor neuron progenitor marker and improves adhesion and proliferation of motor neuron-like cells.

OBJECTIVES: Cell therapy has provided clinical applications to the treatment of motor neuron diseases. The current obstacle in stem cell therapy is to direct differentiation of stem cells into neurons in the neurodegenerative disorders. Biomaterial scaffolds can improve cell differentiation and are widely used in translational medicine and tissue engineering. The aim of this study was to compare the efficiency of two-dimensional with a three-dimensional culture system in their ability to generate functional motor neuron-like cells from adipose-derived stem cells. MATERIALS AND METHODS: We compared motor neuron-like cells derived from rat adipose tissue in differentiation, adhesion, proliferation, and functional properties on two-dimensional with three-dimensional culture systems. Neural differentiation was analyzed by immunocytochemistry for immature (Islet1) and mature (HB9, ChAT, and synaptophysin) motor neuron markers. RESULTS: Our results indicated that the three-dimensional environment exhibited an increase in the number of Islet1. In contrast, two-dimensional culture system resulted in more homeobox gene (HB9), Choline Acetyltransferase (ChAT), and synaptophysin positive cells. The results of this investigation showed that proliferation and adhesion of motor neuron-like cells significantly increased in three-dimensional compared with two-dimensional environments. CONCLUSION: The findings of this study suggested that three-dimension may create a proliferative niche for motor neuron-like cells. Overall, this study strengthens the idea that three-dimensional culture may mimic neural stem cell environment for neural tissue regeneration.

Anatomical Variations of the Musculocutaneous and Median Nerves: A Case Report.

The musculocutaneous nerve is a large terminal branch of the lateral cord of the brachial plexus. It passes under the pectoralis minor and penetrates the coracobrachialis muscle, descending between the biceps brachii and brachialis muscles in the arm. After dissection in upper extremities in a 28-year-old male cadaver, the median and musculocutaneous nerve were found to have variations on the right side where the musculocutaneous nerve formed communications with the median nerve. The median nerve innervated muscles of the front of the arm in this cadaver. In addition, the musculocutaneous nerve did not pierce the coracobrachialis muscle on the right side. Knowledge of these variations is extremely important when planning a surgery in the region of axilla.

Attenuation of Morphine Withdrawal Syndrome by Prosopis Farcta Extract and Its Bioactive Component Luteolin in Comparison with Clonidine in Rats.

BACKGROUND Today, the plant Prosopis farcta is frequently used for traditional medicinal purposes. The aim of this study was the identification of luteolin in P. farcta extract (PFE) and to evaluate its effect on morphine discontinuation syndrome in rats. MATERIAL AND METHODS Using high-performance liquid chromatography (HPCL), luteolin was evaluated in PFE. The frequency of behavioral symptoms of morphine withdrawal (jumping, rearing, and teeth chattering) induced by naloxone challenge were illustrated in morphine-dependent rats receiving PFE, luteolin, saline, or clonidine. LD50 of PFE and luteolin was 540 mg/kg and 150 mg/kg, respectively. Signs of behavioral morphine withdrawal in rats were significantly inhibited by chronic co-administration of PFE, luteolin, or clonidine with morphine. RESULTS This study showed that PFE was less effective than clonidine at a dose of 100 mg/kg, and at doses of 200 mg/kg and 300 mg/kg it was comparable to clonidine, and did not show a significant difference in the reduction of morphine withdrawal symptoms. Luteolin was comparable in 30 mg/kg, 60 mg/kg, and 90 mg/kg with clonidine to reduce the frequency of morphine withdrawal symptoms. PFE can be used as a source of luteolin. CONCLUSIONS The study findings suggest that PFE and luteolin might reduce the signs of narcotic withdrawal. Due to a similar effect to clonidine, its mechanism of action might be through the protein kinase A pathway and might have human therapeutic potential.

Effects of Vitamin D Deficiency on Incidence Risk of Gestational Diabetes Mellitus: A Systematic Review and Meta-analysis.

INTRODUCTION: Proper nutrition is important for overall health, and it reduces healthcare costs associated with malnutrition. Many studies have investigated vitamin D deficiency and its role in gestational diabetes and controversial data have reported. A comprehensive consideration of articles in this field provides the possibility of a general study of this relationship. This meta-analysis is an evaluation of the relationship between vitamin D deficiency and gestational diabetes. MATERIAL AND METHODS: Different databases (such as PubMed, Science Information Institute, EmBase, Scopus, and the Cochrane Library) were searched for studies and eligible English articles published before February 2017 that have reported the risk of gestational diabetes in relation to vitamin D deficiency. This relationship was measured using odds ratios (ORs) with a confidence interval (CI) of 95%. The influence of each study was measured through sensitivity analysis. Funnel plots, Egger regression tests, and the Begg-Mazumdar correlation test were used to determine bias or publication bias. STATA (version 11.2) was used for all analyses. RESULTS: Twenty-six studies were selected as eligible for this research and included in the final analysis. In general, vitamin D deficiency among mothers may be related to an increased risk of gestational diabetes (OR = 1.18; 95% CI, 1.01-1.35; p < 0.001). The serum level of 25(OH)D is less meaningful in people with gestational diabetes than in those who have normal glucose tolerance. Subgroup analysis showed that the results concerning this association may vary with study design but do not change with country of origin. CONCLUSION: Some evidence has shown that vitamin D deficiency may increase the risk of gestational diabetes.

Transdifferentiation of Human Dental Pulp Stem Cells Into Oligoprogenitor Cells.

INTRODUCTION: The nerve fibers in central nervous system are surrounded by myelin sheet which is formed by oligodendrocytes. Cell therapy based on oligodendrocytes and their precursors transplantation can hold a promising alternative treatment for myelin sheet repair in demyelinating diseases. METHODS: Human Dental Pulp Stem Cells (hDPSCs) are noninvasive, autologous and easy available source with multipotency characteristics, so they are in focus of interest in regenerative medicine. In the present study, hDPSCs were differentiated into oligoprogenitor using glial induction media, containing Retinoic Acid (RA), basic Fibroblast Growth Factor (bFGF), Platelet-Derived Growth Factor (PDGF), N2 and B27. The differentiated Oligoprogenitor Cells (OPCs) were evaluated for nestin, Olig2, NG2 and O4 using immunocytochemistry. Also, the expression of nestin, Olig2 and PDGFR-α gens (neuroprogenitor and oligoprogenitor markers) were investigated via RT-PCR technique. RESULTS: The results indicate that glial differentiation medium induces the generation of oligoprogenitor cells as revealed via exhibition of specific glial markers, including Olig2, NG2 and O4. The expersion of nestin gene (neuroprogenitor marker) and Olig2 and PDGFR-α genes (oligoprogentor markers) were detected in treated hDPSCs at the end of the induction stage. CONCLUSION: hDPSCs can be induced to transdifferentiate into oligoprogenitor cells and respond to the routinely applied regents for glial differentiation of mesanchymal stem cells. These data suggest the hDPSCs as a valuable source for cell therapy in neurodegenerative diseases.

Fracture risk in patients with type 2 diabetes mellitus and possible risk factors: a systematic review and meta-analysis.

AIM: Patients with type 2 diabetes mellitus (T2DM) have an increased risk of bone fractures. A variable increase in fracture risk has been reported depending on skeletal site, diabetes duration, study design, insulin use, and so on. The present meta-analysis aimed to investigate the association between T2DM with fracture risk and possible risk factors. METHODS: Different databases including PubMed, Institute for Scientific Information, and Scopus were searched up to May 2016. All epidemiologic studies on the association between T2DM and fracture risk were included. The relevant data obtained from these papers were analyzed by a random effects model and publication bias was assessed by funnel plot. All analyses were done by R software (version 3.2.1) and STATA (version 11.1). RESULTS: Thirty eligible studies were selected for the meta-analysis. We found a statistically significant positive association between T2DM and hip, vertebral, or foot fractures and no association between T2DM and wrist, proximal humerus, or ankle fractures. Overall, T2DM was associated with an increased risk of any fracture (summary relative risk =1.05, 95% confidence interval: 1.04, 1.06) and increased with age, duration of diabetes, and insulin therapy. CONCLUSION: Our findings strongly support an association between T2DM and increased risk of overall fracture. These findings emphasize the need for fracture prevention strategies in patients with diabetes.

The Effects of Dietary Supplements of Calcium, Vitamin D and Estrogen Hormone on Serum Levels of OPG and RANKL Cytokines and their Relationship with Increased Bone Density in Rats.

INTRODUCTION: Osteoprotegerin (OPG)-Receptor activator of nuclear factor kappa-B ligand (RANKL) pathway is one of the contributing factors in the regulation of osteogenesis and bone resorption routes. AIM: The purpose of this study was to evaluate the effects of various dietary supplements on this pathway. MATERIALS AND METHODS: The samples for this study (24 newborn rats) were divided in three groups according to the experiment applied for each group. Rats were given special diet according to their group plan for six weeks. Blood samples were collected to measure their serum levels of OPG and RANKL and all organs of rats were used to measure their bone density too. The results were analysed using appropriate statistical analysing tests. RESULTS: Levels of whole-body bone mineral density in calcium plus vitamin D plus Estrogen (Ca + D + E) group and calcium plus vitamin D (Ca + D) group were significantly increased compared to control group. Mineral density was highest in calcium plus vitamin D plus Estrogen group and was about 0.1357 g/cm2. RANKL had a significant decrease in calcium plus vitamin D plus Estrogen group compared to control and calcium plus vitamin D groups. There was a significant increase in the mean calcium and OPG in both experimental groups rather than control. Also, significant increase in estrogen was observed in Ca + D group than the control group. CONCLUSION: The results showed that intake of calcium and vitamin D and estrogen at determined dose led to an increase in OPG and RANKL cytokines reduction which ultimately led to an increase in bone mineral density. But Ca, D and E synergies were more effective in increasing bone mineral density compared to only the use of Ca and D.

Melatonin improve the sperm quality in forced swimming test induced oxidative stress in nandrolone treated Wistar rats.

This study investigates the effects of melatonin on the sperm quality and testis weight after the combination of swimming exercise and nandrolone decanoate (DECA). Two groups of male Wistar rats were treated for eight weeks as follows; group A consist of CO (control), Sham, N (DECA), S (swimming) and NS (DECA plus swimming); and group B: Sham M (sham melatonin), M (melatonin), MN (melatonin plus DECA), MS (melatonin plus swimming), MNS (melatonin, DECA plus swimming). The motility of sperm was significantly improved in melatonin groups in comparison to N, S and NS groups (P≤0.05).  The left testes weight was decreased in N, NS and MNS groups, and the right testes weight was decreased in N,S,NS, MS and MNS groups in compare with the control group. This study concluded that melatonin probably could improve the sperm motility and sex organs weight after the combination of DECA and exercise.