Retinoic acid and/or progesterone differentiate mouse induced pluripotent stem cells into male germ cells in vitro.

Numerous reagents were employed for differentiating induced pluripotent stem cells (iPSCs) into male germ cells; however, the induction procedure was ineffective. The aim of this study was to improve the in vitro differentiation of mice iPSCs (miPSCs) into male germ cells with retinoic acid (RA) and progesterone (P). miPSCs were differentiated to embryoid bodies (EBs) in suspension with RA with or without progesterone for 0, 4, and 7 days. Then, the expression of certain genes at different stages of male germ cell development including Ddx4 (pre meiosis), Stra8 (meiosis), AKAP3 (post meiosis), and Mvh protein was examined in RNA and/or protein levels by real-time polymerase chain reaction or flow cytometry, respectively. The Stra8 gene expression increased in the RA groups on all days. But, expression of this gene declined in RA + P groups. In addition, an increased expression of Ddx4 gene was observed on day 0 in the P group. Also, a significant upregulation was observed in the expression of AKAP3 gene in the RA + P group on days 0 and 4. However, gene expression decreased in P and RA groups on day 7. The expression of Mvh protein significantly increased in the RA group on day 7. The Mvh expression was also enhanced in the P group on day 4, but it decreased on day 7, while this protein upregulated on day 0 and 7 in the RA + P group. The miPSCs have the capacity for in vitro differentiation into male germ cells by RA and/or progesterone. However, the effects of these inducers depend on the type of combination and an effective time.

Retinoic acid and 17ß-estradiol improve male germ cell differentiation from mouse-induced pluripotent stem cells.

This research aimed to explore the impacts of retinoic acid (RA)/17ß-estradiol (E) induction and embryoid body formation to enhance differentiation of mouse-induced pluripotent stem cells (miPSCs) into male germ cells in vitro. Flow cytometry and qPCR were conducted to describe miPSCs differentiation process. Various temporal expression profiles of germ cell-related genes were traced. Stra8 gene expression increased in the RA group on the 4th day compared to other groups. The RA group experienced a more significant increase than E group. The expression of Sycp3 increased in RA + E group on 4th day compared with other groups. Expression of AKAP3 enhanced in the RA + E group than other groups on day 4. Moreover, miPSCs showed that this gene expression in the RA + E group was increased in comparison to RA and E groups on day 7. AKAP3 gene expression on day 7 of miPSCs decreased in RA and E groups. Flow cytometry data indicated that 3%-8% of the cells in sub-G1 stage were haploid after RA and E induction compared to other groups on day 4. This study showed that miPSCs possess the power for differentiating into male germ cells in vitro via formation of embryoid body by RA with/or E induction.

Deep skin wound healing potential of lavender essential oil and licorice extract in a nanoemulsion form: Biochemical, histopathological and gene expression evidences.

Cutaneous wound healing is one of the public health interests. This study aimed to investigate the effects of nanoemulsion cream containing lavender essential oil and licorice extract on the healing of deep skin wound in a rat model. Eighty-five male Wistar rats were randomly divided into five groups including untreated defects as negative control and defects treated with vehicle ointment, lavender essential oil and licorice extract in emulsion and nanoemulsion forms, and phenytoin 1% as the positive control with an excisional wound on the dorsal neck of each rat. On days 2, 7 and 14 oxidative stress factors were evaluated in wound tissue homogenates. The expression of transforming growth factor-ß (TGF-ß), and type I and type III collagen genes were evaluated. Also, wound tissue samples were processed for Hematoxylin & Eosin and Masson-Trichrome staining. Nanoemulsion reduced the wound area more than other groups significantly. Real-time PCR data demonstrated that nanoemulsion and phenytoin groups have shown the best result in increasing TGF-ß1, Type I and type III collagen genes expression compared to the other groups. Reduction in lipid peroxidation level and increasing in SOD and GPx activity was also significant in the nanoemulsion and phenytoin groups. The formation of granular tissue likewise the appearance of collagen in nanoemulsion and phenytoin groups were faster than the other groups. Nanoemulsion cream containing lavender essential oil and licorice extract exhibited a promising wound healing potential towards the excisional wound model in rats.

Prenatal exposure to noise stress: anxiety, impaired spatial memory, and deteriorated hippocampal plasticity in postnatal life.

Sound pollution is known as an annoying phenomenon in modern life. Especially, development of organisms during fetal life is more sensitive to environmental tensions. To address a link between the behavioral and electrophysiological aspects of brain function with action of hypothalamus-pituitary-adrenal (HPA) axis in stressed animals, this study was carried out on the male Wistar rats prenatally exposed to sound stress. Groups of pregnant rats were exposed to noise stress for 1, 2, and 4 hour(s). The degree of anxiety and the spatial memory were evaluated by elevated plus maze and Morris water maze, respectively. Basic synaptic activity and long-term potentiation (LTP) induction were assessed in the CA3-CA1 pathway of hippocampus. The serum level of corticosterone was measured in the pregnant mothers and the offspring. The behavioral experiments appeared that the stressed animals performed considerably weaker than the control rats. The prenatal stress negatively affected the basic synaptic responses and led to a lower level of LTP. The pregnant animals showed an increased serum corticosterone in comparison with the nonpregnant females. Also the offspring exposed to the noise stress had a more elevated level of corticosterone than the control rats. Our findings indicate that the corticosterone concentration changes markedly coincides the results of behavioral and electrophysiological experiments. We conclude that, similar to other environmental stresses, the sound stress during fetal life efficiently disturbs both cognitive abilities and synaptic activities. The changes in action of HPA axis may contribute to problems of the brain function in the prenatally stress exposed animals.

Vitamin D supplementation restores suppressed synaptic plasticity in Alzheimer's disease.

OBJECTIVES: Hippocampus, an appropriate area of brain for assessment of long-term potentiation (LTP), has been found to be susceptible to neural damages caused by Alzheimer's disease. Evidence indicates that vitamin D supports nerve transmission and synaptic plasticity. Vitamin D receptors are expressed in the hippocampus. METHODS: The present study evaluates occurrence of LTP in the control (CON) group fed with normal regimen and, three groups of Aß-treated rats taking normal (ALZ), vitamin D-free (ALZ - D), or 1,25(OH)2D3 supplemented (ALZ + D) food regimens. In in vivo experiments pre- and post-tetanus field extracellular postsynaptic potentials (fEPSPs) were recorded in the CA3-CA1 pathway. RESULTS: We found that the amplitude of baseline fEPSPs was significantly lower in the ALZ group compared with the CON one; lack of vitamin D further declined the amplitude of responses in the ALZ - D animals. While the tetanic stimulation elicited a considerable LTP in the CON rats it was failed to induce LTP in the ALZ animals. Furthermore, the tetanus considerably depressed the amplitude of recordings in the ALZ - D group. 1,25(OH)2D3 supplementation restored post-tetanus potentiation of fEPSPs amplitude in the ALZ + D groups. DISCUSSION: The present findings signify the crucial role of vitamin D on the basic synaptic transmission and synaptic plasticity.

Effect of postnatal environmental enrichment on LTP induction in the CA1 area of hippocampus of prenatally traffic noise-stressed female rats.

Early-life stress negatively alters mammalian brain programming. Environmental enrichment (EE) has beneficial effects on brain structure and function. This study aimed to evaluate the effects of postnatal environmental enrichment on long-term potentiation (LTP) induction in the hippocampal CA1 area of prenatally stressed female rats. The pregnant Wistar rats were housed in a standard animal room and exposed to traffic noise stress 2 hours/day during the third week of pregnancy. Their offspring either remained intact (ST) or received enrichment (SE) for a month starting from postnatal day 21. The control groups either remained intact (CO) or received enrichment (CE). Basic field excitatory post-synaptic potentials (fEPSPs) were recorded in the CA1 area; then, LTP was induced by high-frequency stimulation. Finally, the serum levels of corticosterone were measured. Our results showed that while the prenatal noise stress decreased the baseline responses of the ST rats when compared to the control rats (P < 0.001), the postnatal EE increased the fEPSPs of both the CE and SE animals when compared to the respective controls. Additionally, high-frequency stimulation (HFS) induced LTP in the fEPSPs of the CO rats (P < 0.001) and failed to induce LTP in the fEPSPs of the ST animals. The enriched condition caused increased potentiation of post-HFS responses in the controls (P < 0.001) and restored the disrupted synaptic plasticity of the CA1 area in the prenatally stressed rats. Likewise, the postnatal EE decreased the elevated serum corticosterone of prenatally stressed offspring (P < 0.001). In conclusion, the postnatal EE restored the stress induced impairment of synaptic plasticity in rats' female offspring.

Neuroprotective effects of alpha-lipoic acid on radiation-induced brainstem injury in rats.

Background and purpose: Alpha-lipoic acid (ALA) is an antioxidant with radioprotective properties. We designed the current work to assess the neuroprotective function of ALA in the presence of oxidative stress induced by radiation in the brainstem of rats. Experimental approach: Whole-brain radiations (X-rays) was given at a single dose of 25 Gy with or without pretreatment with ALA (200 mg/kg BW). Eighty rats were categorized into four groups: vehicle control (VC), ALA, radiation-only (RAD), and radiation + ALA (RAL). The rats were given ALA intraperitoneally 1 h before radiation and killed following 6 h, thereafter superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and total antioxidant capacity (TAC) in the brainstem were measured. Furthermore, a pathological examination was carried out after 24 h, 72 h, and five days to determine tissue damage. Findings/Results: The findings indicated that MDA levels in the brainstem were 46.29 ± 1.64 µM in the RAD group and decreased in the VC group (31.66 ± 1.72 µM). ALA pretreatment reduced MDA levels while simultaneously increasing SOD and CAT activity and TAC levels (60.26 ± 5.47 U/mL, 71.73 ± 2.88 U/mL, and 227.31 ± 9.40 mol/L, respectively). The greatest pathological changes in the rat's brainstems were seen in RAD animals compared to the VC group after 24 h, 72 h, and 5 days. As a result, karyorrhexis, pyknosis, vacuolization, and Rosenthal fibers vanished in the RAL group in three periods. Conclusion and implications: ALA exhibited substantial neuroprotectivity following radiation-induced brainstem damage.

Evaluating the Effects of Probiotic Supplementation on Neuropathic Pain and Oxidative Stress Factors in an Animal Model of Chronic Constriction Injury of the Sciatic Nerve.

Introduction: Neuropathic pain is a common and painful somatosensory nervous system disease, and its treatment remains a medical challenge. Evidence demonstrates that gut microbiota alters in neuropathic pain and, therefore, improvement of the gut flora may affect the disease. The present study aimed to evaluate the antinociceptive effect of probiotics in neuropathic pain and oxidative biomarkers' responsiveness to the probiotic treatment. Methods: Using chronic constriction injury (CCI) of the rats' sciatic nerve, neuropathic pain was induced. Investigating the analgesic effect of the probiotics mixture, 40 male rats were randomly assigned to 4 groups (n=10 for each): Sham-operated (SM), and CCI model rats orally received 1 mL saline (CS), or 100 mg/kg gabapentin (CG) or 1 mL probiotics mixture (CP) Lactobacillus plantarum, Lactobacillus delbrueckii, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Bifidobacterium bifidum (109 CFU of each) daily. Using behavioral tests, the pain was assessed on days 1, 4, 7, 14, and 21 of the study. Finally, the biochemical evaluation of sciatic nerve tissue was done. Results: Probiotics decreased cold and mechanic allodynia and thermal hyperalgesia. Reducing lipid peroxidation levels and increasing total antioxidant capacity, superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity were also significant in the probiotics group. Conclusion: These findings suggest that probiotics have analgesic effects on the CCI model of neuropathic pain via increasing the antioxidant capacity of the rats' sciatic nerve. Highlights: Oral probiotics mixture diminishes cold and mechanical allodynia in chronic constriction injury (CCI) rats.Probiotics mixture reduces thermal hyperalgesia in CCI rats as well as gabapentine.Balancing in oxidant/anti-oxidant system is key pathway in neuropathic pain reduction. Plain Language Summary: Currently, neuropathic pain is an underestimated socioeconomic health problem affecting millions of people worldwide. Neuropathic pain occurs when a health condition affects the nerves that carry sensations to the brain. With neuropathic pain, the pain isn't typically triggered by an event or injury. Instead, the body sends pain signals to the brain unprompted. Neuropathic pain tends to get worse over time. As neuropathic pain could result from an imbalance in the oxidative/antioxidative system, antioxidant supplementation may be a treatment option. Oxidative stress is known to result in the occurrence of molecular mechanisms of diabetes, atherosclerosis, inflammatory bowel disease, and damage to the heart, brain, or transplanted organs. Probiotics are made of good live bacteria and/or yeasts that naturally live in the body and have various health benefits. Consumption of probiotics alone or foods supplemented with probiotics may reduce cell oxidative damage. Incorporating probiotics in foods can provide an excellent strategy to supply dietary antioxidants. This study subjected rats to sciatic nerve ligation to induce neuropathic pain. The oral probiotics mixture was administered for 21 days post-surgery. The result showed that the probiotics mixture administration could reduce oxidative stress and pain in neuropathic pain rats. Therefore, probiotics can prevent and treat many systemic diseases in animal and human studies.

Anti-nociceptive effect of black seed oil on an animal model of chronic constriction injury.

Background and purpose: Traditionally, Nigella sativa L. has been known as a medical intervention to treat numerous diseases. This study aimed at investigating the antihyperalgesic effect of black seed oil (BSO) in an experimental model of neuropathic pain. Experimental approach: Chronic constriction injury (CCI) was performed under anesthesia. The sciatic nerve was ligated with four loose ties. Two separate protocols were used to administer BSO. In chronic treatment, rats were given daily doses of BSO (250, 500, and 1000 mg/kg p.o.) from the 1st day until the 21st post-CCI day. While, in acute treatment, BSO (250, 500, and 1000 mg/kg p.o.) was administered only on the 7th, 14th, and 21st days. CCI and sham groups were given almond oil according to the same schedule. Behavioral scores were determined by evaluation of the paw withdrawal in the plantar, Von Frey, and acetone tests, on the 7th, 14th, and 21st days. Findings/Results: Our results showed that CCI leads to significant allodynia and hyperalgesia in the ipsilateral paw after surgery. Chronic administration of BSO (500 and 1000 mg/kg) obviously attenuated heat hyperalgesia and mechanical allodynia. However, daily administration of BSO did not alter cold allodynia. Nevertheless, when BSO was administered, 30 min before the pain assessment tests, hypersensitivity was not improved in the treated animals. Conclusion and implications: These results demonstrated BSO can inhibit neuropathic pain progression and suggests a potential use of BSO to manage hyperalgesia and allodynia. However, additional research is necessary to approve BSO effectiveness, in neuropathic pain conditions.

Radioprotective effects of N-acetylcysteine on rats' brainstem following megavoltage X-irradiations.

PURPOSE: This study aimed to determine the radioprotective effect of N-acetylcysteine (NAC) on the radiation-induced oxidative stress (OS) in the rats' brainstem. MATERIALS AND METHODS: Eighty rats in four identical groups, including vehicle control (VC), irradiation alone (RAD), irradiation with 1 g/kg of NAC treatment (RAN), and NAC treatment without radiation (NAC) were used. Whole-brain irradiation was performed with a single dose of 25 Gy. The rats received the treatments via intraperitoneal (IP) injection 1 h before the irradiation process. Nitric oxide (NO), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (TAC), and glutathione peroxidase (GPx) were measured in the rats' brainstem and compared between the groups. Furthermore, the pathological study was performed to assess tissue damage after 24 h, 72 h, and 5 days of irradiation. RESULTS: The levels of NO and MDA in the brainstem tissue for the RAD group were 60.37 ± 3.35 µmol/L and 45.10 ± 2.48 µM, respectively, which were higher than those of VC group (NO: 30.41 ± 1.83 µmol/L; MDA: 31.02 ± 1.71 µM). The level of SOD, CAT, TAC, and GPx declined in the RAD compared to the VC group. Pre-treatment with NAC decreased the level of NO and MDA and also enhanced the antioxidant activities. The greatest pathological changes in the rats' brainstems were seen in RAD animals compared to the VC group at 24 h, 72 h, and 5 days. Furthermore, the pathological changes were not observed in the NAC group in all the assessed times. CONCLUSION: Based on the results, NAC can decrease the irradiation-induced oxidative stress and pathology damages in the rats' brainstem. It can be concluded that NAC can be an appropriate radioprotection candidate for the human brainstem.

Anesthesia and analgesia for common research models of adult mice.

Anesthesia and analgesia are major components of many interventional studies on laboratory animals. However, various studies have shown improper reporting or use of anesthetics/analgesics in research proposals and published articles. In many cases, it seems "anesthesia" and "analgesia" are used interchangeably, while they are referring to two different concepts. Not only this is an unethical practice, but also it may be one of the reasons for the proven suboptimal quality of many animal researches. This is a widespread problem among investigations on various species of animals. However, it could be imagined that it may be more prevalent for the most common species of laboratory animals, such as the laboratory mice. In this review, proper anesthetic/analgesic methods for routine procedures on laboratory mice are discussed. We considered the available literature and critically reviewed their anesthetic/analgesic methods. Detailed dosing and pharmacological information for the relevant drugs are provided and some of the drugs' side effects are discussed. This paper provides the necessary data for an informed choice of anesthetic/analgesic methods in some routine procedures on laboratory mice.

Effect of topical Areca palm L. hydroalcoholic extract on burn wound healing in rats.

OBJECTIVES: Wound healing is a complex and dynamic process that begins immediately following tissue injury and continues until the wound is completely healed and remodeled. Applying the most effective burn repair techniques is a constant challenge in medicine. Antiulcerogenic and wound healing properties of Areca palm leaves have been validated through various investigations and animal studies. This study aimed to determine the potential for A. palm hydroalcoholic extract to heal burn wounds in rats. MATERIALS AND METHODS: For 14 days, we examined 40 male Wistar albino rats in 5 groups: those receiving 1% silver sulfadiazine cream (reference standard), those receiving eucerin (positive control), and those receiving 5% and 10% ointments of Areca catechu hydroalcoholic extract (treatment groups). No treatment was given to the negative control group. On the dorsal part of the animals' necks, burn wounds were made. After the rats were sacrificed, the wound contraction rate (WCR) was determined, and the wound sites were histopathologically examined. RESULTS: On the 14th day, the WCR was significantly higher in rats treated with A. palm 10% extract ointment than in rats treated with 5% extract, positive or negative control groups (p < 0.001), or rats treated with silver sulphfadiazine (p = 0.01). After applying a 10% extract ointment to burn wound sites, complete healing occurred with only mild tissue inflammation and edema. CONCLUSION: The study's findings indicate that the hydroalcoholic extract of A. palm L. has the ability to expedite the wound healing process. Additional research is necessary to identify the compounds responsible for their wound healing properties and comprehend their action mechanism.

The effect of female sex hormones on Hsp27 phosphorylation and histological changes in prefrontal cortex after tMCAO.

BACKGROUND: Female sex hormones are protective factors against many neurological disorders such as brain ischemia. Heat shock protein like HSP27 is activated after tissue injury. The main purpose of the present study is to determine the effect of a combined estrogen / progesterone cocktail on the morphology of astrocytes, neurons and Hsp27 phosphorylation after cerebral ischemia. METHODS: One hour after the MCAO induction, a single dose of estrogen and progesterone was injected. The infarct volume was calculated by TTC staining 24 h after ischemia. Immunohistochemistry was used to show the effects of estrogen and progesterone on astrocyte and neuron morphology, as well as the Western blot technique used for the quantitation of phosphorylated Hsp27. RESULTS: The combined dose of estrogen and progesterone significantly decreased astrocytosis after ischemia and increased neuron survival. There was a large increase in Hsp27 phosphorylation in the penumbra ischemic region after stroke, which was significantly reduced by hormone therapy. CONCLUSION: Our results indicate that the neuroprotective effect of neurosteroids in the brain may be due to the modulation of heat shock proteins.

Anti-osteoarthritis potential of peppermint and rosemary essential oils in a nanoemulsion form: behavioral, biochemical, and histopathological evidence.

BACKGROUND: This study aimed to evaluate the effect of nanoemulsion containing peppermint and rosemary essential oils in rats with osteoarthritis (OA). METHODS: In this experimental study, we prepared a nanoemulsion containing peppermint and rosemary essential oils by spontaneous emulsification and evaluated the nanoemulsion's dermal irritation and toxicity. Investigating the analgesic effect of the nanoemulsion, we randomly assigned 36 male rats to 6 groups: Control (saline injection into the knee), osteoarthritis (intra-articular injection of 2 mg monosodium iodoacetate), and four groups of OA treated with nanoemulsion gel, nanoemulsion solution, rosemary and peppermint essential oil gel, or diclofenac sodium. Treatments were administered topically at a dose of 1 ml daily. Using behavioral tests, we assessed pain on days 1, 4, 7, and 14 after injection. Finally, we did the histopathological and biochemical evaluation of rats' knee joints. RESULTS: There were no irritation signs on the animals' skin after receiving the nanoemulsion and no changes in the hematological and biochemical parameters of rats' blood compared to the control group. Receiving nanoemulsion decreased the mechanical (P < 0.001) and thermal allodynia (P < 0.05), thermal hyperalgesia (P < 0.05), and ambulatory-evoked pain in comparison with the OA group. Also, the nanoemulsion receiving rats showed an increase in SOD and GPx activity and a decrease in MDA level. Histopathology of synovial tissues confirmed the results of behavioral and biochemical tests. CONCLUSION: The nanoemulsion containing essential oils of peppermint and rosemary reduces osteoarthritis pain via increasing antioxidant capacity and improving the histopathological features of the rats' knee joint.

Light deprivation improves melatonin related suppression of hippocampal plasticity.

In early postnatal life, sensory inputs deeply influence development as well as function of the brain. Plasticity of synaptic transmission including its experimentally induced form, long-term potentiation (LTP), is affected by sensory deprivation in neocortex. This study is devoted to assess if dark rearing and a dark phase synthesized hormone melatonin influence LTP in the hippocampus, an area of brain involved in learning and memory. In vivo experiments were carried out on two groups of 45-days-old male Wistar rats kept in standard 12-h light/dark condition [light reared (LR) tested during the light phase] or in complete darkness [dark reared (DR)] since birth to testing. Each group, in turn, was divided to two, vehicle- and melatonin-treated, groups. Stimulating the Schaffer collaterals of CA3 area of hippocampus extracellular postsynaptic potentials (EPSPs) were recorded in the CA1 area. Having the stable baseline responses to the test pulses, the hippocampus was perfused by either vehicle or 2 microg melatonin and EPSPs were recorded for 30 min. Then, for induction of LTP, the tetanus was applied to the Schaffer collaterals and the field potentials were pooled for 120-min post-tetanus. The light deprivation resulted in a significant augmentation in the amplitude of baseline responses. Also, we observed a melatonin-induced increase in amplitude of the baseline recordings in either LR or DR animals. Tetanic stimulation elicited LTP of EPSPs in both LR and DR groups, robustly in the former where it lasted for about 90 min. Generally, melatonin inhibited the production of LTP in the two groups especially in the LR animals leading to a noticeable depression. We concluded that higher level of neuronal activity in the DR rats gives rise to a lower level of LTP. Weaker effect of melatonin on blocking the potentiation of post-tetanus EPSPs in the DR rats may be the result of a desensitization of melatonin receptors due to chronically increased levels of this hormone in the visually deprived rats.

Environmental enrichment restores impaired spatial memory and synaptic plasticity in prenatally stress exposed rats: The role of GABAergic neurotransmission.

Evidence shows that prenatal stress negatively affects cognitive functions and activity of neuronal circuits in postnatal age. Environmental enrichment counteracts deficits induced by early life stress. We examined if behavioural function and synaptic plasticity are sensitive to prenatal stress and, how much environmental enrichment and GABAergic system impact these phenomena. Animals were exposed to noise stress during the third trimester of foetal life. Groups of the stressed animals remained intact (S-SH) or received enrichment (S-EE) from postnatal day 22 for one month. Also, two groups received either saline (S-SH-S) or bicuculline (S-SH-B). One enriched group received muscimol (S-EE-M). The control groups were intact (C-SH), enriched (C-EE), or received bicuculline (C-SH-B) or saline (C-SH-S). We assessed learning and memory and, hippocampal long-term potentiation (LTP). Serum corticosterone levels were detected as a measure of stress condition. We found that stress reduced spatial performance and suppressed LTP in the S-SH animals. Postnatal enrichment restored both spatial learning and memory and synaptic plasticity in the S-EE rats. GABAergic antagonism strengthens maze performance and LTP induction in the S-SH-B group. However, muscimol prevented the positive effects of enrichment in the S-EE-M animals. Environmental enrichment and GABAergic modulation may improve disrupted spatial performance and synaptic plasticity.

Behavioral and electrophysiological aspects of cognition in neonate rats lactated by morphine addicted mothers.

OBJECTIVES: In addition to genetic factors, environmental phenomena during postnatal age highly affect development and, in turn, function of the brain. The present work evaluates if morphine consumption during lactation period influences the spatial performances and synaptic plasticity in rats at neonatal period of age. MATERIALS AND METHODS: Three groups of mothers were subcutaneously administered by 5 (M5), 10 (M10) or 20 (M20) mg/kg morphine every 12 hours during the lactation period. At 45 days old, their offspring were introduced to Morris water maze for assessment of spatial learning and memory. Basic field excitatory post-synaptic potentials (fEPSPs) were recorded in the CA1 area of hippocampus and, then, long term potentiation (LTP) was induced by tetanic stimulation. RESULTS: We found that the M10 and M20 rats spent more time and traveled longer distance to find the hidden platform of maze when compared to the control animals (P<0.05 for all comparisons). Similarly, these two morphine-exposed groups were inferior in the memory consolidation compared to their control counterparts. Comparing control and M20 rats revealed that morphine exposure decreases the mean amplitude and slope 10-90% of fEPSPs about 30 percent (P<0.001 for both comparisons) and inhibits the LTP induction in the CA1 area circuits. CONCLUSION: The present study provides behavioral and electrophysiological proofs for negative effect of morphine on the hippocampal-related function in the neonatally morphine-exposed rats.

The protective effect of bone marrow mesenchymal stem cells in a rat model of ischemic stroke via reducing the C-Jun N-terminal kinase expression.

Ischemic stroke is the main cause of disability and mortality worldwide. Apoptosis and inflammation have an important role in ischemic brain injury. Mesenchymal stem cells (MSCs) have protective effects on stroke treatment due to anti-inflammatory properties. The inhibition of the C-Jun N-terminal kinase (JNK) pathway may be one of the molecular mechanisms of the neuroprotective effect of MSCs in ischemic brain injury. Twenty-eight male Wistar rats were divided randomly into 3 groups. Except the sham group, others subjected to transient middle cerebral artery occlusion (tMCAO). Bone marrow MSCs or saline were injected 3 h after tMCAO. Sensorimotor behavioral tests were performed 24 and 72 h after ischemia and reperfusion (I/R). The rats were sacrificed 72 h after I/R and infarct volume was measured by TTC staining. The number of apoptotic neurons and astrocytes in the peri-infarct area was assessed by TUNEL assay. The morphology of cells was checked by Nissl staining, and the expression of p-JNK was detected by immunohistochemistry and Western blot. Behavioral scores were improved and infarct volume was reduced by MSCs 24 h and 72 h after tMCAO. TUNEL assay showed that neuronal apoptosis and astroglial activity in the penumbra region were reduced by MSCs. Also, Nissl staining showed lower neuronal apoptosis in BMSCs-treated rats compared to controls. JNK phosphorylation which was profoundly induced by ischemia was significantly decreased after MSCs treatment. We concluded that anti-apoptotic and anti-inflammatory effects of MSCs therapy after brain ischemia may be associated with the down-regulation of p-JNK.

The Wound Healing Effects of Herbal Cream Containing Oliveria Decumbens and Pelargonium Graveolens Essential Oils in Diabetic Foot Ulcer Model.

BACKGROUND: The number of diabetic patients in adult population is increasing. All this population are at risk of developing diabetic foot ulcers (DFUs) that are associated with unwanted ailments and high mortality. In spite of current therapies for DFUs, further therapies are needed to help the patients. METHODS: The efficacy of herbal cream containing Pelargonium graveolens and Oliveria decombens essential oils was evaluated topically for treatment of DFUs in rat animal model in comparison with two other herbal formulas containing each essential oil alone, placebo (the basic formula without active ingredients) and normal saline as control groups. After anesthesia of diabetic rats (n=75) induced by streptozotocin (STZ), diabetic wounds were visible on the hind dorsal surface of the foot. The treatments were initiated on Day 1 and repeated 3 times a day for thirteen consecutive days. On day 1, 3, 5, 8 and 13, the wound sizes were determined and assessed histologically. RESULTS: Three herbal formulations reduced the size of wounds in rats with DFUs, while the cream containing combined herbals of O. decumbens and P. graveolens essential oils had the highest tissue repair in DFU rat models. CONCLUSION: Due to better wound healing effects of combined herbal cream containing O. decumbens and P. graveolens essential oils, it can be recommended in treatment of DFUs.

Estrogen agonist genistein differentially influences the cognitive and motor disorders in an ovariectomized animal model of Parkinsonism.

OBJECTIVES: Parkinson's disease (PD) is a progressive neurological disorder associated with motor disabilities and cognitive dysfunction as well. Evidence indicates that PD occurs less frequently in women than men, confirming a role for steroid hormones in protection of dopaminergic nigrostriatal neurons. It is reported that soy genistein, an estrogen agonist phytoestrogen, display neuroprotective effects against neuronal death. In this study we evaluated the effect of genistein in animal models of Parkinsonism (P) and Parkinsonism + ovariectomized (OP). MATERIALS AND METHODS: The experiments were carried out on the control, P and OP animals. Learning and memory abilities were evaluated using Morris water maze. The latency and speed of locating the platform were measured as cognitive indices. Motor behaviors were assessed by testing the animals in rota rod and the latency to fall from the rod was scored. RESULTS: We found that Parkinsonism leads to the cognitive and motor disabilities; ovariectomy intensified these disorders. Whereas genistein treatment improved the maze performances in both P and OP animals it failed to influence the kinetic problems. Genistein displayed a neuroprotective effect on dopaminergic neurons. CONCLUSION: Positive impact of genistein on the spatial learning and memory may reflect its effects on the nigrostriatal pathway and striatum. Nevertheless, ineffectiveness of genistein on the motor disorders, despite its neuroprotective impacts, led us to conclude that the cognitive improvement by genistein may also contribute to its effects in other areas of brain.