Behavioral, histopathological, and biochemical evaluations on the effects of cinnamaldehyde, naloxone, and their combination in morphine-induced cerebellar toxicity.

Long-term morphine use for therapeutic approaches may lead to serious side effects. Several studies have suggested opioid antagonist and antioxidant therapy for reducing adverse effects of morphine. Cinnamaldehyde has a potent anti-oxidant property. In this study, separate and combined effects of cinnamaldehyde and naloxone (an opioid receptor antagonist) on behavioral changes and cerebellar histological and biochemical outcomes were investigated after long-term morphine administration. Seventy-eight rats were divided into two major morphine-treated and morphine-untreated groups. Morphine-treated group was subdivided into seven subgroups for receiving vehicle, normal saline, cinnamaldehyde (1.25, 5, and 20 mg/kg), naloxone, and cinnamaldehyde plus naloxone before morphine. Morphine-untreated group was subdivided into six subgroups and treated with vehicle, cinnamaldehyde (1.25, 5, and 20 mg/kg), naloxone, and their combination. Chemical compounds were administered for 28 consecutive days. Behavioral tests including footprint, rotarod, and beam balance tests were employed. Histopathological and biochemical alterations of cerebellum were determined. Body and cerebellum weights, stride width, time spent on the rotarod, Purkinje cell number, thickness of molecular and granular layers, superoxide dismutase (SOD), and total antioxidant capacity (TAC) decreased as a result of administrating morphine. Morphine increased beam transverse time, malondealdehyde (MDA), tumor necrosis factor-α (TNF-α), and caspase-3 levels. Histopathological changes such as cellular vacuolation and loss were also produced as a result of treatment with morphine. Cinnamaldehyde, naloxone, and their combination treatments improved all the above-mentioned alterations induced by morphine. We concluded that cinnamaldehyde produced a neuroprotective effect through anti-oxidant, anti-inflammatory, apoptotic, and probably naloxone-sensitive opioid receptor interaction mechanisms.

Comparison of the effects of hydroalcoholic extract of Capparis spinosa fruit, quercetin and vitamin E on monosodium glutamate-induced toxicity in rats.

Capparis spinosa L. has many biological effects such as antioxidant properties. In the present study, we compared the effects of the hydro-alcoholic extract of Capparis spinosa fruit, quercetin (Q), and vitamin E (Vit E) on monosodium glutamate (MSG)-induced toxicity. The following groups were designed: Control groups (normal saline and/or corn oil); MSG group (4.00 g kg-1 MSG); MSG + low dose extract group (4.00 g kg-1 MSG with 100.00 mg kg-1 extract); MSG + high dose extract (HDE) group (4.00 g kg-1 MSG with 300.00 mg kg-1 extract); MSG + Q group (4.00 g kg-1 MSG with 10.00 mg kg-1 Q); MSG + Vit E group (4.00 g kg-1 MSG with 200.00 mg kg-1 Vit E). All chemicals were orally administered for 14 consecutive days. Tissue specimens from the heart, kidney, and liver tissues and blood samples were collected for histopathological and biochemical evaluations. The results showed that the MSG-induced tissue edema, congestion, and inflammatory cell infiltration were resolved by HDE, Q, and Vit E treatments. These chemicals also restored tissue malondialdehyde level and superoxide dismutase activity. Besides, alterations induced by MSG in serum levels of aspartate transaminase, alanine aminotransferase, urea, lactate dehydrogenase, and creatine kinase-MB were also resolved. It is concluded that Capparis spinosa fruit extract, Q and Vit E can produce approximately similar protective effects on tissue function through oxidative stress alleviation and antioxidant mechanisms restoration.

Effects of rabbit pinna-derived blastema cells on tendon healing.

OBJECTIVES: Tendon healing is substantially slow and often associated with suboptimal repair. Cell therapy is one of the promising methods to improve tendon repair. Blastema, a population of undifferentiated cells, represents characteristics of pluripotent mesenchymal stem cells and has the potentials to be used in regenerative medicine. The aim of this study was to investigate the use of blastema allotransplantation in rabbit tendon healing. MATERIALS AND METHODS: In this study, one rabbit was used as a blastema donor, and twenty-four rabbits were divided into control and treatment groups. Blastema cells were obtained from ear pinna upon punch hole injury in the donor rabbit. Under general anesthesia, a complete transverse tenotomy was performed on the midsubstance of deep digital flexor tendon followed by suture-repair. In the treatment group, 1 × 106 blastema cells suspended in buffer saline were injected intratendinously at the repair site, while the control group received only the buffer saline. Cast coaptation was maintained for two weeks. Eight weeks after the operation, tendons were harvested, and histopathological, biomechanical, and biochemical assays were performed on samples. RESULTS: Mechanical testing showed a significant increase in ultimate load, energy absorption, stiffness, yield load, stress, and strain in blastema-treated tendons compared to controls. Also, higher hydroxyproline content and improved collagen alignment along with lower inflammatory cell infiltration and decreased angiogenesis were observed in blastema-treated tendons. CONCLUSION: Increased levels of hydroxyproline and improved histopathological and biomechanical parameters in the treatment group suggest that blastema cells could be considered an adjunct to tendon repair in rabbits.

Surgical management of penile sarcoid in a stallion.

This report describes surgical management and breeding implications of a case of penile sarcoid associated with penis laceration in a 4-year-old Kurdish stallion. A large fleshy mass on the distal end of the penis that resulted in urethral meatus deviation and dysuria was detected in a physical examination. No evidence of local extent or metastasis was detected. Under general anaesthesia, the involved distal portion of the penis was removed through partial phallectomy. Histopathological examination of the mass confirmed a fibroblastic sarcoid. Partial phallectomy was successful for management of penile sarcoid and resulted in no postoperative complications or tumour recurrence in long-term follow up; however, successful ejaculation and semen collection have not been achieved.

Safranal, a constituent of saffron, exerts gastro-protective effects against indomethacin-induced gastric ulcer.

AIMS: Several natural products have been evaluated for management of gastric ulcer induced by non-steroidal anti-inflammatory drugs. Safranal, a plant-derived chemical, has a potent antioxidant and anti-inflammatory properties. The present study was aimed to evaluate possible gastro-protective effects of safranal against indomethacin-induced gastric ulcer in rats. Lansoprazole (a proton pump inhibitor) was used as a reference drug. MATERIALS AND METHODS: Thirty rats were divided into five groups. Groups 1 and 2 received vehicle. Groups 3, 4 and 5 treated with 0.063, 0.25 and 1 mg/kg safranal. Group 6 received 30 mg/kg lansoprazole. All groups except of group 1 received indomethacin (50 mg/kg) ingestion. Six hours later, animals were euthanized and their stomachs were removed. Gastric contents volume and pH were measured. Gastric ulcer area and protective index were evaluated using image J software. Histological changes were evaluated by light microscope. Malondialdehyde (MDA) level, superoxide dismutase (SOD) activity, total antioxidant capacity (TAC) content, tumor necrosis factor-alpha (TNF-α) and Caspase-3 levels were determined in the gastric tissue. KEY FINDINGS: Safranal and lansoprazole normalized gastric volume and pH, reduced gastric ulcer area and produced gastric protection. Indomethacin-induced histological changes and tissue biochemical alterations were ameliorated by the above-mentioned treatments. SIGNIFICANCE: The results of the present study suggest the involvement of anti-secretory, anti-oxidant, anti-inflammatory and anti-apoptotic mechanisms in gastro-protective effect of safranal. In addition, gastro-protective effect of safranal was comparable to lansoprazole.

Crocin exerts improving effects on indomethacin-induced small intestinal ulcer by antioxidant, anti-inflammatory and anti-apoptotic mechanisms.

Crocin is a plant-derived carotenoid and bears potent antioxidant property. Ranitidine (a histamine H2 receptor blocker) is used for peptic ulcer treatment. The present study was planned to investigate the effects of crocin and ranitidine on indomethacin-induced ulcer in small intestine of rats. Animals were randomized into two major groups including indo-methacin (10.00 mg kg-1, ulcer group, 48 rats) and normal saline (1.00 mL kg-1, intact group, 48 rats) groups. Each of these two major groups was subdivided into eight subgroups for intra-peritoneal (IP) injections of normal saline, crocin (2.50, 10.00 and 40.00 mg kg-1), ranitidine (5.00 and 20.00 mg kg-1), crocin (2.50 and 10.00 mg kg-1) plus ranitidine (5.00 mg kg-1). Indomethacin induced intestinal ulcer was characterized by bleeding, inflammation, epithelial hyperplasia and crypt loss. This non-steroidal anti-inflammatory drug (NSAID), indomethacin decreased goblet cell number and superoxide dismutase (SOD) activity and increased small intestine weight, organo-somatic index (OSI), malodealdehyde (MDA), tumor necrosis factor-α (TNF-α) and caspase-3 contents of intestine. Crocin resolved all the above-mentioned parameter changes induced by indomethacin. These treatments produced no significant effects on the above-mentioned parameters of intact group. The results of the present study showed tissue protective and anti-ulcer effects of crocin on small intestine by antioxidant, anti-inflammatory and anti-apoptotic mechanisms. Ranitidine alone showed no effect; however, in combination with crocin it exerted recovery effects. It is recommended that crocin, be considered as a therapeutic agent for NSAIDs-induced intestinal damage management.

Carnosine improves functional recovery and structural regeneration after sciatic nerve crush injury in rats.

AIMS: Peripheral nerve injury represents a substantial clinical problem with insufficient or unsatisfactory treatment options. Current researches have extensively focused on the new approaches for the treatment of peripheral nerve injuries. Carnosine is a naturally occurring pleotropic dipeptide and has many biological functions such as antioxidant property. In the present study, we examined the regenerative ability of carnosine after sciatic nerve crush injury using behavioral, biochemical, histological and ultrastructural evaluations. MATERIALS AND METHODS: Seventy-two rats were divided into six groups including control, sham, crush and carnosine (10, 20 and 40 mg/kg) groups. Crush injury in left sciatic nerve was induced by a small haemostatic forceps. Carnosine was administered for 15 consecutive days after induction of crush injury. Sciatic functional index (SFI) was recorded weekly. Histopathological and ultrastructural evaluations were made using light and electron microscopes, respectively. Sciatic nerve tissue malondialdehyde (MDA), superoxide dismutase (SOD) and tumor necrosis factor-alpha (TNF-α) levels were measured. Gastrocnemius muscle weight was determined. KEY FINDINGS: Carnosine at the doses of 20 and 40 mg/kg accelerated SFI recovery. Wallerian degeneration severity and myelinated fibers density, myelin sheath thickness and diameter as well as ultrastructural changes of myelinated axons were improved. It also recovered nerve tissue biochemical (MDA, SOD and TNF-α) changes induced by crush injury. Muscle weight ratio was reached to near normal values. Our results suggest a regenerative effect of carnosine. Inhibition of oxidative stress and inflammatory pathways, along with provocation of myelination and prevention of muscular atrophy might be involved in this effect of carnosine. SIGNIFICANCE: Carnosine treatment might be considered as a therapeutic agent for peripheral nerve regeneration and its functional recovery.

The effects of safranal, a constitute of saffron, and metformin on spatial learning and memory impairments in type-1 diabetic rats: behavioral and hippocampal histopathological and biochemical evaluations.

Safranal is one of saffron constituents and has antioxidant and neuroprotective properties. Metformin is used as an anti-diabetic drug. This study was planned to investigate the separate and combined treatment effects of safranal and metformin on diabetes-induced learning and memory impairments by behavioral and hippocampal histopathological and biochemical evaluations. Diabetes was induced by intraperitoneal injection of streptozotocin (STZ), treatments with safranal (0.025, 0.1 and 0.4 mg/kg), metformin (50 and 200 mg/kg), and a combination of low doses of this chemicals were initiated after confirmation of diabetes and continued for 37 days. Blood glucose concentration was measured before and on days 15, 25 and 35 after injection of streptozotocin. Learning and memory tested using Morris Water Maze (MWM) on days 40-45 and on day 45 hippocampal specimens were collected for determination of malodialdehyde (MDA), tumor necrosis factor-alpha (TNF-α) and Caspase-3 levels and superoxide dismutase (SOD) activity. The hippocampus was also designed for light microscopy evaluation. Hyperglycemia, spatial learning and memory impairments, hippocampal neuron loss, increase of hippocampal MDA, TNF-α and caspase-3 levels and decrease of SOD activity were observed in diabetic rats. Safranal (0.1 and 0.4 mg/kg), metformin (200 mg/kg) and safranal (0.025 mg/kg) with metformin (50 mg/kg) improved the above-mentioned behavioral, histopathological and biochemical changes. Safranal and metformin and their combination improved learning and memory impairments in STZ-induced diabetic rats. Antioxidant, anti-inflammatory and antiapoptotic mechanisms might be involved. It is recommended that safranal be considered for diabetes management.

Fabrication of novel tubular scaffold for tendon repair from chitosan in combination with zinc oxide nanoparticles.

Chitosan bears numerous properties, such as biocompatibility, biodegradability and non-toxicity making it suitable for use in different biomedical fields. Zinc (Zn) is required for fibroblasts proliferation and collagen synthesis as essential elements of wound healing. Its nanoparticles are well known for their capability to enhance wound healing by cell adhesion and migration improvement through growth factors-mediated mechanisms. Poor blood supply and unique histological characteristics of tendon make its regeneration always slow. Also, adhesion formation between tendon and its surrounding tissues is another problem for neotendon to return to its normal structure and functional activities. In this study, a novel tubular scaffold of zinc oxide (ZnO) nanoparticles loaded chitosan has been fabricated for tendon repair. Experimental complete tenotomy of deep digital flexor tendon in a rabbit model was done and scaffolds were placed in the transected area after two ends suturing. After four and eight weeks, adhesion formation around the tendons and tissue reaction to the scaffolds were evaluated macroscopically. Inflammation, angiogenesis and collagen fibers arrangement were also analyzed in histopathological evaluations. After eight weeks, the scaffolds were absorbed completely, adhesions around the tendon were decreased and there was no sign of significant tissue reaction and/or infection in histopathological analyses. The reduced adhesion formation, improved gliding function and better histopathological characteristics suggest this scaffold application as a potential therapy in treatment of tendon acute injuries.

The effects of bone marrow-derived mesenchymal stem cells on ovalbumin-induced allergic asthma and cytokine responses in mice.

OBJECTIVES: Allergic Asthma is an inflammatory disease of the lungs that is characterized by increased infiltration of leukocytes into the airways, limiting the respiratory function. Studies suggest that a defective general regulatory system against inflammation could be a significant factor in allergic asthma. It has been shown that Mesenchymal stem cells (MSCs) have a cellular immunosuppressive therapeutic potential for inflammatory disorders. We investigated whether administration of MSCs during allergen challenge would affect the underlying mechanisms in allergic airways inflammation. MATERIALS AND METHODS: Fifty mice were used in five control and experimental groups; the experimental mice sensitized by intraperitoneal injection of OVA and aluminum hydroxide emulsion on days 0, 7, and 14, were then challenged intranasally with OVA or sterile PBS on days 14, 25, 26, and 27. Before allergen challenge on day 14, experimental mice received tail vein injection of MSCs in PBS, whereas control mice received PBS alone. Cytokine and IgE analyses were carried out using lung washes as well as serum samples. RESULTS: Our, results showed that MSCs significantly reduced total cells and eosinophilia and serum OVA-specific IgE concentration in OVA-sensitized and challenged mice. Also, results showed that MSCs markedly inhibited expressions of Th2 and Th17 cytokines and elevated levels of Treg cytokines. CONCLUSION: we found that administration of MSCs could be used as a potential therapeutic approach for allergic asthma.

Effects of silver nanoparticles on Staphylococcus aureus contaminated open wounds healing in mice: An experimental study.

The microorganisms have been noted as the main cause of delayed wound healing. The most common pathogen causing the wound infections is Staphylococcus aureus. Silver nanoparticles (AgNPs) show ample antibacterial activities. In the present study, the effect of AgNPs on mouse wounds inoculated with S. aureus was investigated. Sixty male mice (20 to 30 g) were anesthetized, full-thickness skin wounds were made on their back and then the bacterial suspension was added to each wound bed. Treatments were administered on wound bed topically including gentamicin (8 mg kg-1), AgNPs (0.08 mg kg-1, 0.04 mg kg-1 and 0.02 mg kg-1) and normal saline in the control group. Wound healing was monitored macroscopically by taking digital photographs on days 0, 7, 14 and 21 of the experiment. Topical application of gentamicin and AgNPs (0.08 and 0.04 mg kg-1) significantly increased the rate of wound healing more than treatment with AgNPs at a dose of 0.02 mg kg-1and normal saline. The presence of silver nanoparticles in AgNPs groups (especially 0.08 mg kg-1) improved wound appearance better than other groups without silver nanoparticles (gentamicin and control groups) and led to lesser wound scars. According to data analysis, healing rate of treated mice with gentamicin and AgNPs (0.08 mg kg-1) was significantly (p < 0.001) faster than treated mice with other AgNPs doses and normal saline. The results of current study introduced an in vivo nanosilver accelerating effects on the treatment of on S. aureus infected skin wounds.

Effects of histidine and vitamin C on isoproterenol-induced acute myocardial infarction in rats.

In the present study, we investigated the effects of histidine and vitamin C (alone or in combination) treatments against isoproterenol (a ß-adrenergic receptor agonist)-induced acute myocardial infarction in rats. We used propranolol (a ß-adrenergic receptor blocker) to compare the results. Rats were given intraperitoneal injections of histidine (40 mg kg(-1)) and vitamin C (40 mg kg(-1)) alone and combined daily for 21 days. Propranolol (10 mg kg(-1)) was orally administered daily for 10 days (from day 11 to day 21). Myocardial infarction was induced by subcutaneous injections of 150 mg kg(-1) of isoproterenol at an interval of 24 hr on days 20 and 21. Blood and tissue samples were taken for histopathological and biochemical evaluations following electrocardiography recording on day 21. Isoproterenol elevated ST segment, increased heart weight, heart rate, serum activities of aspartate transaminase, lactate dehydrogenase, creatine kinase-MB and heart tissue content of malondialdehyde, and decreased R wave amplitude and superoxide dismutase and catalase activities of heart tissue. Necrosis, edema and inflammatory cells infiltration were observed in myocardial tissue sections. Our results indicated that histidine and vitamin C alone, and especially in combination prevent isoproterenol-induced cardiotoxicity and have similar protective effects with propranolol. Cardioprotective effects of histidine and vitamin C may be associated with their ability to reduce free radical-induced toxic effects.

The effects of crocin, insulin and their co-administration on the heart function and pathology in streptozotocin-induced diabetic rats.

OBJECTIVE: Crocin is a saffron constituent with a potent anti-oxidant activity. The present study investigated the effects of crocin and insulin treatments (alone or in combination) on cardiac function and pathology in diabetic rats. MATERIALS AND METHODS: Diabetes was induced by intraperitoneal (i.p.) injection of streptozotocin (STZ, 50 mg/kg). Thereafter, crocin (5, 10 and 20 mg/kg, i.p.), subcutaneous (s.c.) injection of insulin (4 IU/kg) and their combination were administered for eight weeks. Blood glucose level and whole heart and body weights were measured. Electrocardiography (ECG) was carried out using the lead II. Serum concentrations of lactate dehydrogenase (LDH), creatine kinase-MB isoenzyme (CK-MB), and the heart tissue malodialdehyde (MDA) and superoxide dismutase (SOD) contents were determined. The heart lesions were evaluated by light microscopy. RESULTS: STZ decreased body weight and increased whole heart weight/body weight ratio. It also decreased heart rate, and increased RR and QT intervals and T wave amplitude. STZ increased blood glucose, serum LDH and CK-MB levels, augmented heart tissue MDA content, decreased SOD content of heart tissue, and produced hemorrhages, degeneration, interstitial edema, and fibroblastic proliferation in the heart tissue. Crocin (10 and 20 mg/kg, i.p.), insulin (4 IU/kg, s.c.) and their combination (5 mg/kg of crocin with 4 IU/kg of insulin) treatments recovered the ECG, biochemical and histopathological changes induced by STZ. CONCLUSION: The results showed cardioprotective effects of crocin and insulin in STZ-induced diabetic rats. The antioxidant and anti-hyperglycemic properties of crocin and insulin may be involved in their cardioprotective actions.

Histopathological and behavioral evaluations of the effects of crocin, safranal and insulin on diabetic peripheral neuropathy in rats.

OBJECTIVES: Crocin and safranal, the major constituents of saffron, exert neuroprotective effects. In the present study, we investigated the effects of crocin and safranal (alone or in combination with insulin) on peripheral neuropathy in diabetic rats. MATERIALS AND METHODS: Diabetes was induced by intraperitoneal (i.p.) injection of 60 mg/kg of streptozotocin (STZ) and confirmed by blood glucose level higher than 250 mg/dl. After confirmation of diabetes, crocin (30 mg/kg, i.p.), safranal (1 mg/kg, i.p.) (alone or in combination with insulin) and insulin (5 IU/kg, s.c.) were administered for eight weeks. Neuropathic pain was evaluated using acetone drop test. Histopathological changes of sciatic nerve were evaluated using light microscope. Blood glucose levels and sciatic nerve malondialdehyde (MDA) contents were also measured. RESULTS: STZ caused cold allodynia, edema and degenerative changes of sciatic nerve, hyperglycemia and an elevation of sciatic nerve MDA levels. Crocin, safranal and insulin improved STZ-induced behavioral, histopathological and biochemical changes. Combined treatments produced more documented improving effects. CONCLUSION: The results of the present study showed neuroprotective effects of crocin, safranal and insulin in a rat model of diabetic neuropathy. In addition, crocin and safranal enhanced the neuroprotective effect of insulin. The neuroprotective effects of theses chemical compounds could be associated with their anti-hyperglycemic and antioxidant properties.

Hawthorn ethanolic extracts with triterpenoids and flavonoids exert hepatoprotective effects and suppress the hypercholesterolemia-induced oxidative stress in rats.

OBJECTIVES: 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 hypercholesterolemia-increased 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 hypercholesterolemia-induced oxidative stress and hepatic injuries. Moreover, the hypocholesterolemic effect of extracts was found comparable to orlistat.

Effects of microinjection of histamine into the anterior cingulate cortex on pain-related behaviors induced by formalin in rats.

BACKGROUND: The present study was aimed to investigate the effects of microinjection of histamine and its H1, H2 and H3 receptor antagonists, mepyramine, ranitidine and thioperamide, respectively, into the anterior cingulate cortex (ACC) on pain-related behaviors induced by formalin in rats. METHODS: Two stainless steel guide canulas were bilaterally implanted into the ACC of anaesthetized rats. For induction of pain, intraplantar (ipl) injection of a 2.5% formalin solution was performed. The duration of paw licking/biting and the number of paw flinching were recorded in 5 min blocks for 60 min. Locomotor activity was assessed using an open-field test. RESULTS: Formalin produced a marked biphasic pattern of pain. Histamine reduced the second phases of paw licking/biting and flinching. Mepyramine (2 µg/side) prevented the suppressive effect of histamine (1 µg/side) on second phase of pain, but at a dose of 8 µg/side it did not inhibit the suppressive effects of 4 µg/side of histamine. Ranitidine at doses of 2 and 8 µg/side prevented histamine (1 and 4 µg/side)-induced antinociception. Thioperamide not only suppressed the second phases of pain, but also increased the suppressive effect of histamine. Naloxone prevented suppressive effects of histamine and thioperamide on pain. Mepyramine (8 µg/side) suppressed locomotor activity. CONCLUSION: The results of the present study showed pain suppressing effects for histamine. Histamine H2 and H3, and to a lesser extent, H1 receptors might be involved in histamine-induced antinociception. Opioid receptors might be involved in suppressive effects of histamine and thioperamide.

The role of nicotinic acetylcholine and opioid systems of the ventral orbital cortex in modulation of formalin-induced orofacial pain in rats.

Nicotinic acetylcholine and opioid receptors are involved in modulation of pain. In the present study, we investigated the effects of microinjection of nicotinic acetylcholine and opioid compounds into the ventral orbital cortex (VOC) on the formalin-induced orofacial pain in rats. For this purpose, two guide cannulas were placed into the left and right sides of the VOC of the brain. Orofacial pain was induced by subcutaneous injection of a diluted formalin solution (50µl, 1.5%) into the right vibrissa pad and face rubbing durations were recorded at 3-min blocks for 45min. Formalin produced a marked biphasic pain response (first phase: 0-3min and second phase: 15-33min). Epibatidine (a nicotinic receptor agonist) at doses of 0.05, 0.1 and 0.2µg/site, morphine (an opioid receptor agonist) at doses of 0.5, 1 and 2µg/site and their sub-analgesic doses (0.025µg/site epibatidine with 0.25µg/site morphine) combination treatment suppressed the second phase of pain. The antinociceptive effect induced by 0.2µg/site of epibatidine, but not morphine (2µg/site), was prevented by 2µg/site of mecamylamine (a nicotinic receptor antagonist). Naloxone (an opioid receptor antagonist) at a dose of 2µg/site prevented the antinociceptive effects induced by 2µg/site of morphine and 0.2µg/site of epibatidine. No above-mentioned chemical compounds affected locomotor activity. These results showed that at the VOC level, epibatidine and morphine produced antinociception. In addition, opioid receptor might be involved in epibatidine-induced antinociception, but the antinociception induced by morphine was not mediated through nicotinic acetylcholine receptor.

Effects of histidine and n-acetylcysteine on experimental lesions induced by doxorubicin in sciatic nerve of rats.

In this study, the effect of separate and combined intraperitoneal (i.p.) injections of histidine and n-acetylcysteine were investigated on experimental damage induced by doxorubicin (DOX) in sciatic nerve of rats. DOX was i.p. injected at a dose of 4 mg/kg once weekly for four weeks. Histidine and n-acetylcysteine were i.p. injected at a same dose of 20 mg/kg. Cold and mechanical allodynia were recorded using acetone spray and von Frey filaments tests, respectively. The sciatic nerve damage was evaluated by light microscopy. Plasma levels of malondialdehyde (MDA) and total antioxidant capacity (TAC) were measured. Histidine and especially n-acetylcysteine at a same dose of 20 mg/kg suppressed cold and mechanical allodynia, improved sciatic nerve lesions and reversed MDA and TAC levels in DOX-treated groups. Combination treatment with histidine and n-acetylcysteine showed better responses when compared with them used alone. The results of the present study showed peripheral neuroprotective effects for histidine and n-acetylcysteine. Reduction of free radical-induced toxic effects may have a role in neuroprotective properties of histidine and n-acetylcysteine.

A novel dispersive micro solid phase extraction using zein nanoparticles as the sorbent combined with headspace solid phase micro-extraction to determine chlorophenols in water and honey samples by GC-ECD.

This study presents a new technique, dispersive micro solid phase extraction (DMSPE) combined with headspace solid phase micro-extraction (HS-SPME) for extraction and determination of chlorophenols (CPs) in water and honey samples using a Gas Chromatography-Electron Capture Detector (GC-ECD). Zein nanoparticles were made by liquid-liquid dispersion and applied for the first time as the sorbent phase in DMSPE. In the proposed DMSPE-HS-SPME method, 1% w/v of ethanolic zein solution was added to an aqueous sample and then a dose of the in-situ generated zein nanoparticles was applied to a pre-concentration of target analytes. Thermal desorption of analytes was performed after the isolating sorbent phase, and then HS-SPME was applied for enrichment prior to introducing to gas chromatography. All the important parameters influencing efficiency of the extraction process such effects of salt, pH, sorbent concentration, temperature, sorbent solution volume in DMSPE procedure, extraction temperature, extraction time, desorption temperature and time in the HS-SPME procedure were investigated and optimized. Results showed that under optimum extraction conditions, detection limits (signal to noise ratio=3) were in the range of 0.08-0.6 ng mL(-1) and evaluations for relative standard deviations (RSDs %) were between 6.62% and 8.36%.