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Pluripotent stem cells derived from testis is a new, natural, and unlimited source for cell therapy in regenerative medicine and represent a possible alternative to replacing of all cells in the body. Here, we designed a simple co-culture system of spermatogonia cells with Sertoli cells for the generation of embryonic stem-like cells from mouse testis. The importance of our simple method will be clear when we compared it with other complex and time-consuming methods. Embryonic stem-like colonies with sharp border confirmed by real-time PCR, immunocytochemistry and flow cytometry assessments. Embryonic stem-like colonies were immunopositive for pluripotency markers. Transition of spermatogonia cells to embryonic stem-like cells was accompanied by extensive changes in gene expression. These changes included significant increase in pluripotency genes expression and significant decrease in germ cell-specific genes expression. Also, we proved the differentiation capacity of embryonic stem-like cells to neuroepithelial-like cells which were immunoreactive to Nestin and Neurofilament 68. Evaluation of genes expression during in vitro differentiation into neuroepithelial-like cells showed high-level expression of Nestin whether this gene approximately has no expression in undifferentiated embryonic stem-like cells. Also, expression of pluripotency genes has significantly decreased in neuroepithelial-like cells compared with embryonic stem-like cells. This study shows that embryonic stem-like cells derived from testis are capable to differentiate into neuroepithelial-like cells that may provide a cellular reservoir usable for neurodegenerative disorders.

Background: New research proposes the pluripotency of spermatogonial cells obtained from testis. These spermatogonia-derived stem cells are called embryonic stem-like cells that express embryonic stem cell markers and differentiate to the three germ layers., Objectives: The aim of the present study was to generate embryonic stem-like cells from neonatal mouse testis., Materials and Methods: The Testis cells were collected from neonatal mouse. After decapsulation, testis was mechanically dissected and dissociated via a two-step mechanical and enzymatic digestion. The spermatogonia and sertoli cells were cultured together in Dulbecco's modified Eagle's medium (DMEM) supplemented with 15% FBS and LIF. Before one week, several small spermatogonia colonies were observed on top of the monolayer of sertoli cells. These colonies were passaged every four days. ES-Like cells colonies that resembled ES cell was appeared within 2-3 weeks (at passages 5). Real time PCR was performed to analyze the expression of a subset of pluripotency markers, as well as germ cell-specific genes. ES Like cells were confirmed with SSEA1, SOX2 and Oct4 immunofluorescence stainng as pluripotency markers., Results: The Results showed that at fifth passages, the pluripotency genes; Nanog and c-myc have significant increase in ES-Like cells in compare with spermatogonia cells, whereas the spermatogonial markers; Stra8, mvh, and piwill2 became downregulated. In addition to these pluripotency genes, the ES cell marker SSEA-1, SOX2 and Oct4 were expressed in the ES-like cells, similar to ES cells., Conclusions: This researh indicates pluripotency evidence of ES-like cells derived from testis. ES-like cells shows some molecular characteristics with embryonic stem cells.

Co(III) salen complex with N,N'-dipyridoxyl (1,4-butanediamine) Schiff-base ligand as tetradentate ligand was synthesized and characterized by the elemental and spectroscopic analysis. The interaction of this complex with calf thymus DNA (ct DNA) has been investigated in vitro using UV absorption, fluorescence spectroscopy, thermal denaturation and gel electrophoresis techniques. The binding constant has been estimated to be 1×10(4)M(-1) using UV absorption. The addition of ct DNA to Co(III) salen solution resulted in a fluorescence quenching. The binding constant and site size binding have been calculated in connection with other experimental observations show that the interactive model between Co(III) salen and ct DNA is an intercalative one. The interaction between plasmid DNA (pTZ57R DNA) and this complex is confirmed by gel electrophoresis studies. Furthermore, the interaction between HSA and Co(III) salen complex was investigated by UV absorption, fluorescence spectroscopy and molecular modeling. The binding constant for the interaction of this complex with HSA were found to be 3.854×10(4)M(-1) using UV absorption, which was in good agreement with the binding constant obtained from fluorescence method (3.866×10(4)M(-1)). The binding distance between HSA and this complex was estimated to be 2.48nm according to Förster theory of non-radioactive energy transfer. Molecular modeling studies suggested that hydrophobic interaction was the predominant intermolecular forces stabilizing Co(III) complex-HSA system., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Zymoseptoria is a newly described genus that includes the prominent wheat pathogen Zymoseptoria tritici (synonyms Mycosphaerella graminicola and Septoria tritici). Studies indicated that the center of origin of Z. tritici is in the Middle East where this important pathogen emerged during the domestication of wheat. Several Zymoseptoria species have been found on uncultivated grasses in the Middle East, and in this article we describe two new Zymoseptoria species from Iran. These species, isolated from Elymus repens, Dactylis glomerata and Lolium perenne, are named Z. ardabiliae and Z. pseudotritici. Both species were identified by means of morphological characteristics and phylogenetic analyses of a seven-gene DNA dataset. These taxa comprise some of the closest known relatives of the wheat pathogen Z. tritici, confirming the reported close phylogenetic relationship between Z. tritici and Z. pseudotritici.

Unlabelled: BACKGROUND AND THE PURPOSE OF STUDY: Concerning the different effects of essential oils from Nepeta genus on the central nervous system including pain killing effect, this study was designed to evaluate the antinociceptive and anti-inflammatory effects of essential oil of Nepeta pogonosperma Jamzad et Assadi (NP), a recently identified species., Methods: Air-dried aerial parts of NP were hydrodistillated and GC-MS analysis of obtained essential oil was conducted. Total 24 male Wister rats weighing 225 ± 25 gm were studied. Essential oil of NP was administered intraperitoneally at the doses of 50 mg/kg, 100 mg/kg and 200 mg/kg for the experimental groups. Control rats received equal volume (2 ml/kg) of normal saline. Antinociception was assessed by tail flick test (after 30 minutes) and formalin test (for further 60 minutes). Then the animal was sacrificed and the paw edema was measured using a water plethysmometer., Results: 4aα,7α,7aβ-nepetalactone and 1,8-cineole were found as the main concentrated components of NP essential oil. All the doses of NP showed antinociception. NP 200 mg/kg reduced the pain sensation in tail flick (p <0.01) and formalin test (p <0.001 in both phases). In paw edema test, NP 100 and 200 mg/kg significantly reduced the inflammation (p <0.01 and p <0.05)., Conclusion: This study reveals that the essential oil of NP may minimize both the acute and chronic forms of nociception and may have potent role against inflammation, but the dose should be maintained precisely to obtain the intended effect.

Induction of demyelination in the central nervous system induce the oligodendrocyte progenitors to proliferate, migrate, and differentiate for restoring new myelin sheathes around demyelinated axons. Factors which increase the response of endogenous progenitor cells could be used to improve remyelination. In the current study, the effect of bFGF on lysolecithin-induced demyelination and remyelination processes in mouse optic chiasm and nerves was investigated. Lysolecithin was injected into the optic chiasm of Balb/C mice. Two groups of animals received doses of bFGF (1 or 5 ng/kg i.p.) just before and every 3 days after lysolecithin injection. Delay and amplitude of visual evoked potential (VEP) waves were recorded as indices of axonal demyelination at 7th, 13th, and 28th days post-lesion. Myelin basic protein (MBP) and Olig2 gene expressions were studied as indices of myelination and oligodendrocyte precursors' recruitment into the lesion. Lysolecithin elongated delay of P1 wave and declined the amplitude of P1-N1 wave. Lysolecithin decreased MBP and increased Olig2 expression in different days post-lesion. Lysolecithin-induced changes in VEPs were partially ameliorated by endogenous repair. bFGF reduced the increased delay, increased the reduced amplitude of P1-N1 wave, increased MBP gene expression, and accelerated the increasing pattern of Olig2. bFGF seems to be able to potentiate the endogenous repair mechanisms of myelin. Its effect on demyelination and remyelination processes seems to be mediated by oligodendrocyte progenitor cells and their differentiation to myelinating cells.

Electromagnetic fields (EMFs) may affect the endogenous neural stem cells within the brain. The aim of this study was to assess the effects of EMFs on the process of toxin-induced demyelination and subsequent remyelination. Demyelination was induced using local injection of lysophosphatidylcholine within the corpus callosum of adult female Sprague-Dawley rats. EMFs (60 Hz; 0.7 mT) were applied for 2 h twice a day for 7, 14, or 28 days postlesion. BrdU labeling and immunostaining against nestin, myelin basic protein (MBP), and BrdU were used for assessing the amount of neural stem cells within the tissue, remyelination patterns, and tracing of proliferating cells, respectively. EMFs significantly reduced the extent of demyelinated area and increased the level of MBP staining within the lesion area on days 14 and 28 postlesion. EMFs also increased the number of BrdU- and nestin-positive cells within the area between SVZ and lesion as observed on days 7 and 14 postlesion. It seems that EMF potentiates proliferation and migration of neural stem cells and enhances the repair of myelin in the context of demyelinating conditions.

Recent studies suggest that normobaric hyperoxia (HO) protects the rat brain from ischemia reperfusion (IR) injury. Protein kinase C (PKC) is a key signaling molecule involved in protection against IR injury but its role in protective effect of HO in brain injury in unknown. In this study we attempted to see if PKC is involved in the effect of HO. Rats were divided into four main experimental groups. The first two were exposed to 95 % oxygen (HO) in a chamber 4 h/day for 6 consecutive days. Each of these groups had a control group exposed to 21 % oxygen. To investigate the role of PKC during HO, chelerythrin chloride (CHEL, 1 mg/kg/day), a PKC inhibitor, or its vehicle was given to animals for 6 days. After 24 h, the rats were subjected to 60 min of right middle cerebral artery occlusion (MCAO). After 24 h reperfusion neurological deficit scores, infarct volume, brain edema and blood-brain Barrier (BBB) permeability were assessed. HO decreased the infarct volume and brain edema in comparison with controls. PKC inhibition was associated with a significant increase in infarct size in both HO and control animals. PKC inhibition was unable to change brain edema in the experimental groups. Both HO and PKC inhibition reduced the BBB permeability within 24 h post occlusion of middle cerebral artery. Although both HO and PKC inhibition were associated with inhibition of BBB permeability during ischemic brain injury in rats, the neuroprotective effect of HO was independent of PKC in the MCAO model.

Drug addiction is an occurrence with physiological, psychological, and social outcomes. Repeated drug exposure causes neuronal adaptations and dependency. It has been shown that CaMKIIα enzyme contributes to morphine dependency. The locus coeruleus nucleus has been implied in the morphine withdrawal syndrome. This research focuses on the behavioral and molecular adaptations that occur in the locus coeruleus neurons in response to the chronic morphine exposure. Adult male Wistar rats were injected by morphine sulfate (10 mg/kg/s.c.) at an interval of 12 h for a period of nine subsequent days. On the tenth day, naloxone (1 mg/kg/i.p.) was injected 2 h after the morphine administration. Somatic withdrawal signs were investigated for 30 min. We concluded that the inhibition of CaMKIIα by administration of KN-93, the specific inhibitor of this enzyme, significantly attenuated some of the withdrawal signs. In molecular method, the expression of CaMKIIα protein has been enhanced in locus coeruleus of the morphine dependent rats. These findings indicate that CaMKIIα may be involved in the modulation of the naloxone-induced withdrawal syndrome, and treatment with KN-93 may have some effects on this system., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Background: Amyloid beta (Aβ) is believed to be responsible for the synaptic failure that occurs in Alzheimer's disease (AD), but there is little known about the functional impact of Aβ on intrinsic neuronal properties. Here, the cellular effect of Aβ-induced neurotoxicity on the electrophysiological properties of CA1 pyramidal neurons and the mechanism(s) of neuroprotection by CB1 cannabinoid receptor activation was explored., Methods: A combination of behavioural, molecular and electrophysiological approaches was used., Results: Bilateral injections of the Aβ peptide fragment (1-42) into the prefrontal cortex caused a significant impairment in the retention and recall capability in the passive avoidance tasks and significantly increased the level of active caspase-3 in the hippocampus. Whole-cell patch clamp recordings revealed a significant reduction in the intrinsic action potential (AP) frequency and an increase in the discharge irregularity in the absence of synaptic inputs in Aβ treated group. Aβ treatment induced also significant changes in both the spontaneous and evoked neuronal responses. However, co-treatment with ACEA, a CB1 receptor agonist, preserved almost the normal intrinsic electrophysiological properties of pyramidal cells., Conclusions: In vivo Aβ treatment altered significantly the intrinsic electrophysiological properties of CA1 pyramidal neurons and the activation of CB1 cannabinoid receptors exerted a strong neuroprotective action against Aβ toxicity., (Copyright © 2012 S. Karger AG, Basel.)

This study was conducted to evaluate the antinociceptive and anti-inflammatory activities of the essential oil of Nepeta crispa. The study was done using the tail-flick and formalin test pain models and the paw oedema model of inflammation. Male Wistar rats were used as the animal model. The essential oil dose-dependently produced analgesia in the acute pain models, including the tail-flick (p < 0.001) and the first phase of the formalin test (p < 0.01). In the late phase of the formalin test, as a model of chronic pain, the essential oil significantly reduced the pain-induced behaviour (p < 0.01). Nepeta crispa essential oil caused potent anti-inflammatory effects in the formalin-induced paw inflammation model and significantly reduced the paw oedema in all applied doses (p < 0.01). Its effects on pain in both the acute and chronic pain models and its anti-inflammatory effect suggest both central and peripheral mechanisms of action for the essential oil obtained from N. crispa.

Effects of morphine on synaptic transmission and plasticity in the hippocampus area CA1 following in vivo sodium salicylate and the potential molecular mechanism were investigated. Population spikes (PS) were recorded from stratum pylamidale of area CA1 following stimulation of Schaffer collaterals in slices taken from control and sodium salicylate injected rats. To induce long term potentiation (LTP), a 100Hz tetanic stimulation was used. Acute in vitro morphine increased baseline PS amplitude in control slices but not in slices taken from sodium salicylate treated rats. In vivo chronic salicylate did slightly decrease and/or destabilize LTP of CA1 synaptic transmission. We also found that mRNA of NR2A subunit of NMDA receptor was reduced in the hippocampus of sodium salicylate treated rats as compared to control ones. Following LTP induction, the mRNA of NR2A and PP1 (protein phosphatase 1) in slices taken from salicylate-treated rats were more than those of control ones. After long-term exposure to in vitro morphine, high frequency stimulation (HFS) decreased NR2A mRNA level significantly in sodium salicylate treated slices. It is concluded that in vivo sodium salicylate causes tolerance to excitatory effect of morphine and changes the ability of HFS to induce PS LTP in the hippocampus area CA1 in vitro. These changes in synaptic response may be due to alterations in NR2A and PP1 expression., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Multiple sclerosis (MS) is a demyelinating disease that affects the central nervous system. MS is the most common neurological disorder in young adults with a greater incidence among females. Male gonadal hormones have a protective effect on neural system development and myelin maturation. In this study, we investigate the effect of castration on lysolecithin-induced demyelination and remyelination processes using visual evoked potentials, in addition to measuring the expressions of Olig2, MBP, Nogo-A and GFAP mRNAs as oligodendrocyte or astrocyte markers; and histological assessments by myelin-specific staining. We observed more expanded demyelination with delayed repair process in castrated rats. Expression levels of the aforementioned marker genes confirmed histological and electrophysiological observations. Our results showed a pivotal role for endogenous male hormones in the context of demyelinating insults. It may also account for the different prognosis of MS between male and female genders and provide new insights for therapeutic treatments.

Multiple sclerosis frequently affects the optic apparatus, particularly optic chiasm and nerves. Here, we have reported the structural and molecular characteristics of remyelination in the adult rat optic chiasm and nerves. Moreover, considering the proximity of optic chiasm and 3rd ventricle, we have tried to determine if proliferating cells residing in 3rd ventricle region are able to migrate in response to experimental demyelination of the optic chiasm. Following local demyelination by lysolecithin, remyelination pattern in longitude of optic chiasm and proximal nerves was investigated using myelin staining and marker genes expression. Furthermore, cell tracing was carried out using BrdU labeling of proliferating cells prior to gliotoxin injection. Morphometric analysis revealed that demyelination was considerable on days 7 and 14 and an incomplete remyelination occurred on day 28 post-lesion. Interestingly, myelin repair was more evident in the caudal part of chiasm, compared to rostral part and proximal optic nerves. Following chiasm and nerve demyelination, trains of BrdU+ cells were seen near the 3rd ventricle which subsequently moved to lesion site. Nestin was significantly up-regulated in 3rd ventricle surroundings. At the lesion site, Nogo-A gene expression was significantly decreased on days 7 and 14 post lesion, while Olig2, nestin, and GFAP expression was increased on day 7. The changes were then reversed by the time. Myelin repair in optic chiasm seems to be mediated by endogenous progenitors and stem cells. Adult 3rd ventricle proliferating cells may play a role in this context by mobilization into the demyelinated chiasm.

In order to identify a specific marker for T. harzianum AS12-2, a strain capable of controlling rice sheath blight caused by Rhizoctonia solani, UP-PCR was performed using five universal primers (UP) both separately and in pairwise combinations. The application of two UP primers resulted in the amplification of unique fragments from the genomic DNA of T. harzianum AS12-2, clearly distinguishing it from other Trichoderma strains. The unique fragments had no significant sequence homology with any other known sequence available in databases. Based on the sequences of the unique fragments, 14 oligonucleotide primers were designed. Two primer sets amplified a fragment of expected size from the DNA of strain T. harzianum AS12-2 but not from any other examined strains belonging to T. harzianum, to other Trichoderma species assayed, or to other common fungi present in paddy fields of Mazandaran province, Iran. In conclusion, SCAR (sequence characterized amplified regions) markers were successfully identified and rapid, reliable tools were provided for the detection of an effective biocontrol Trichoderma strain, which can facilitate studies of its population dynamics and establishment after release into the natural environment.

Background: This study aims to differentiate human induced pluripotent stem cells (hiPSCs) into oligodendrocyte precursors and assess their recovery potential in a demyelinated optic chiasm model in rats., Methodology/principal Findings: We generated a cell population of oligodendrocyte progenitors from hiPSCs by using embryoid body formation in a defined medium supplemented with a combination of factors, positive selection and mechanical enrichment. Real-time polymerase chain reaction and immunofluorescence analyses showed that stage-specific markers, Olig2, Sox10, NG2, PDGFRα, O4, A2B5, GalC, and MBP were expressed following the differentiation procedure, and enrichment of the oligodendrocyte lineage. These results are comparable with the expression of stage-specific markers in human embryonic stem cell-derived oligodendrocyte lineage cells. Transplantation of hiPSC-derived oligodendrocyte progenitors into the lysolecithin-induced demyelinated optic chiasm of the rat model resulted in recovery from symptoms, and integration and differentiation into oligodendrocytes were detected by immunohistofluorescence staining against PLP and MBP, and measurements of the visual evoked potentials., Conclusions/significance: These results showed that oligodendrocyte progenitors generated efficiently from hiPSCs can be used in future biomedical studies once safety issues have been overcome.

Estradiol is a neuroactive steroid found in several brain areas such as locus coeruleus (LC). It modulates nociception by binding to its receptors and also by allosteric interaction with other membrane-bound receptors like glutamate and GABA(A) receptors. LC is involved in noradrenergic descending pain modulation. In order to study the effect of 17beta-estradiol on both acute and persistent pain modulation and its mechanisms, formalin was injected into the male rat's hind paw. Formalin-induced responses including licking, flexing duration and paw jerking frequency were recorded for 60 min after injection of 50 microl of 2% formalin. The results of the current study showed that intra-locus coeruleus injection of 17beta-estradiol attenuated the second phase, but not the acute phase of formalin-induced pain (P<0.05). AMPA receptor antagonists CNQX had no effect on pain-modulatory effect of 17beta-estradiol. Estrogen and GABA(A) receptor antagonists (ICI 182,780 and bicuculline, respectively) could not reverse the antinociceptive effect of 17beta-estradiol. However, NMDA receptor antagonist APV significantly antagonized the analgesic effect of 17beta-estradiol on flexing behaviour (P<0.05). It may be concluded that the analgesic effect of 17beta-estradiol in formalin-induced inflammatory pain is mediated through interaction with membrane-bound receptors, probably the NMDA receptors., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

Multiple sclerosis (MS) patients may suffer from optic disturbances. Toxin-induced demyelinations have frequently been developed to investigate the cellular and structural aspects of demyelination and remyelination processes, separately. The present study describes functional consequence of lysolecithin (LPC)-induced lesion in the adult rat optic nerves and chiasm by recording the visual evoked potentials (VEPs) from the visual cortex and its correlation with myelin basic protein (MBP) expression in lesion site. Records of VEP were obtained at 2, 7, 14 and 28 days post-injection. We observed that the VEPs generated by light stimuli progressively changed in both amplitude and latency after the lesion as well as in comparison with those generated in control animals. These observations were confirmed through measurement of mRNA expression level for MBP which is one of the important genes expressed in mature oligodendrocytes and Schwann cells. The level of MBP mRNAs in demyelinated chiasm and optic nerves decreased following lysolecithin injection with its least value on day 7, and then it increased to the control level 14 days post-lesion. However, it continued to increase even after that and reached a maximum level 28 days post lesion. Results of the present paper show that, LPC injection in the chiasm share functional and molecular alterations which are found in demyelinating disorders in both the optic nerves and chiasm and also these alterations were coming back to level of control animal on 28 days post lesion, which is typically seen in myelin repair process. The present paper provides new insights into the experimental toxin-induced models that may be useful for evaluating the functional recovery of demyelinated optic nerves and chiasm following various repairing strategies. It also seems to be useful for studying the protective or remyelinating effects of different therapies in e.g. optic apparatus which is more affected by MS., (2010 Elsevier B.V. All rights reserved.)

Cognitive deficits have been observed in patients with multiple sclerosis (MS) due to hippocampal insults. Antioxidant vitamins D and E are suggested for patients suffering from neurodegenerative diseases like MS, while their mechanisms of action are not well understood. Here, we have tried to study the effects of these vitamins on demyelination, cell death, and remyelination of rat hippocampus following local ethidium bromide (EB) injection. Animals received 100 mg/kg vitamin E or 5 microg/kg of vitamin D3 for 2, 7, or 28 days. The extent of demyelination, myelin staining intensity, and expression of myelin basic protein and caspase-3 were investigated using histological and immunoblotting verification. Administration of EB alone caused demyelination, cell death, and afterward an endogenous repair. Vitamins E and D3 reduced the EB-induced damage and increased the endogenous remyelination of hippocampus. Although the anti-apoptotic effect of these vitamins and protection against demyelination were predictable based on their antioxidant effect, our results indicated the positive effect of vitamins E and D3 on process of remyelination by endogenous progenitor cells and supported their possible therapeutic effects in the context of demyelinating diseases like MS.

Essential oil components as result of non host disease resistance of plants have high capability to introduce as alternative of chemical pesticides. Thymus migricus essential oil was selected to investigation of its antifungal activity on survival and growth of Aspergillus flavus. For obtain essential oil first Leaves and flowers of Th. migricus collected then dried. The Essential oil was extracted by means of hydro-distillation and afterwards GC-MS analysis was performed to identify their components. The main constituents that resulted were Thymol (44.9%), Geraniol (10.8%), gamma-Terpinene (10.3%), Citronellol (8.5%) and p-Cymene (7.2%). EC50 and MIC (Minimum Inhibitory Concentration) of Th. migricus oil against A. flavus was 324.42 microl/l and 451.62 microl/l, respectively. Whereas EC50 and MIC for chemical thiabendazol was 650 microl/l and 1635 microl/l, respectively. The EC50 and MIC concentrations of Th. migricus oil in antifungal activity examination were used in aflatoxin inhibition test. Result of HPTLC measurement showed that both of concentrations inhibit aflatoxin production completely compares to control with 7.63 ppm aflatoxin production. In other word, Th. migricus oil can suppress aflatoxin production in concentrations lower than EC50 for mycelium growth.

Increasing attentions have been paid on the application of essential oils and plant extracts for control of postharvest pathogens due to their natural origin and less appearance of resistance in fungi pathogens. Some Aspergillus species are toxigenic and responsible for many cases of food and feed contamination. Some Toxins that produce with some Aspergillus species are known to be potent hepatocarcinogens in animals and humans. The present work evaluated the parameters of antifungal activity of the essential oils of Zataria multiflora, Thymus migricus, Satureja hortensis, Foeniculum vulgare, Carum capticum and thiabendazol fungicide on survival and growth of different species of Aspergillus. Aerial part and seeds of plant species were collected then dried and its essential oils isolated by means of hydrodistillation. Antifungal activity was evaluated in vitro by poisonous medium technique with PDA medium at six concentrations. Results showed that all essential oils could inhibit the growth of Aspergillus species. The essential oil with the best effect and lowest EC50 and MIC (Minimum Inhibitory Concentration) was Z. multiflora (223 microl/l and 650 microl/l, respectively). The chemical composition of the Z. multiflora essential oil was analyzed by GC-MS.

The effects of chronic morphine exposure on synaptic plasticity in the CA1 region of the hippocampal slice preparation using extracellular recordings of the population spike (PS) evoked in response to Schaffer collateral stimulation were studied. High frequency stimulation (HFS; 1X100 Hz) and theta pulse stimulation (TPS; 5 Hz trains for 3 min) were used as patterned activities. The results showed that in rats chronically treated with morphine (dependent group), TPS induced long-term depression (LTD) of PS in CA1 in the absence of in vitro morphine. This TPS-induced PS LTD was blocked in the presence of either AP5 (NMDAR antagonist) or CPX (A1 adenosine receptor antagonist) alone, but was not blocked when AP5 and CPX were co-applied. This TPS-induced PS LTD was also blocked in the presence of either 8-PT (a selective A1 adenosine receptor antagonist) or MRS1220 (a specific A3 receptor antagonist). Additionally, when TPS was applied prior to HFS, PS long-term potentiation (LTP) was blocked. However, when TPS was applied after HFS, there was no reversal of PS LTP in slices from dependent rats in contrast to controls which displayed reversal of LTP. Both the PS LTD and the absence of PS LTP reversal were blocked by in vitro application of morphine. It is concluded that morphine withdrawal was associated with greater depression of CA1 PS elicited by natural stimulus induced activity pattern. This effect was associated with changes in NMDA and adenosine receptors due to chronic morphine administration. Such an in vitro preparation could provide a novel paradigm to investigate withdrawal effects on synaptic plasticity.

Voltage-gated N-type Ca(2+) channels play a central role in regulation of neurotransmitter release. The II-III linker of N-type calcium channel alpha(1B) subunit (exons 17-21 in rats) contains synaptic protein interaction (synprint) site, which plays a pivotal role in efficient neurotransmission. Using RT-PCR analysis of rat hippocampus and lumbar spinal cord transcripts, the expressions of Ca(v)2.2 exon 18a and a novel Ca(v)2.2 splice variant, which were detected in this study, have been investigated, and they showed tissue-specific pattern. The new variant contains a large deletion in II-III linker that produces a stop codon, which is predicted to produce a two-domain-truncated channel. To study the effect of inflammatory pain on the expression pattern of these variants, animals were treated with intraplantar formalin, and the amount of splice variants in lumbar spinal cord was measured. The results showed that chronic inflammatory pain increases the Ca(v)2.2 mRNA levels lacking exon 18a and decreases the amount of full-length variants, which do not have any deletion. Determining different Ca(v)2.2 splice variants in rat nervous system and the impact of inflammatory pain on the splicing pattern suggest a possible regulatory role for calcium channel alternative splicing.

Purpose: Previous studies have shown that the anticonvulsant effects of low-frequency stimulation (LFS) can be affected by activation of adenosine receptors. In the present study, the effect of LFS at different frequencies on kindling rate and adenosine receptors gene expression was investigated., Methods: Animals were kindled by perforant path stimulation in a rapid kindling manner. LFS (0.5, 1, and 5 Hz) was applied after termination of each kindling stimulation. Seizure severity was measured according to behavioral and electrophysiologic parameters. At the end of the experiments, adenosine A(1) and A(2A) receptor gene expression were measured., Results: The inhibitory effect of LFS on kindling acquisition was observed at all frequencies. In addition, the inhibitory action of LFS on enhancement of field excitatory postsynaptic potential slope and population spike amplitude during kindling acquisition was not affected by the LFS frequency. However, the effects of LFS on paired-pulse recordings were greater at frequency of 5 Hz. Application of LFS during kindling acquisition also prevented the kindling induced decrease in the A(1) receptor gene expression and attenuated the level of A(2A) receptor gene expression in the dentate gyrus. These effects were also greater at the frequency of 5 Hz., Discussion: According to these data, it may be suggested that the antiepileptogenic effects of LFS, developed through inhibition of synaptic transmission in the dentate gyrus, is mediated somehow through preventing the decrease of A(1) receptor and through attenuating the A(2A) receptor gene expression. These effects might be dependent on the frequency of LFS.

Chronic morphine leads to dependence, tolerance, and neural apoptosis. Vitamin C inhibits the withdrawal syndrome in morphine-dependent subjects and prevents apoptosis in experimental models. Sodium-dependent vitamin C transporter (SVCT) type-2 is the main transporter for carrying vitamin C into the brain and neural cells. The mechanism(s) by which vitamin C inhibits morphine dependence in not understood. SVCT activity determines the vitamin C availably within the nervous system. We have examined the alterations in the expression of SVCT1, SVCT2, and its splice variants in morphine-tolerant rats. Morphine (20 mg/kg) was injected twice/day to male rats for either 7 or 14 days. The development of analgesic tolerance was assessed using tail-flick test. Lumbar spinal cord and the hippocampus were isolated for RNA extraction. Semiquantitative reverse transcriptase-polymerase chain reaction method was used to assess the levels of gene expression. Administration of morphine for 7 or 14 days reduced the expression level of SVCT2 in both hippocampus and dorsal lumbar spinal cord of rats. SVCT2 expression was reduced in vitamin C-, and vitamin C combined with morphine-treated animals. Results did not show SVCT2 splice variation. SVCT1 did not express in control or morphine-treated rats. It seems that reduced expression level of SVCT2 might be involved in the development of morphine side effects such as tolerance and dependency.

Low frequency stimulation (LFS) has an inhibitory effect on rapid perforant path kindling acquisition. In the present study the role of adenosine A(1) and A(2A) receptors in mediating this inhibitory effect was investigated. Rats were kindled by perforant path stimulation using rapid kindling procedures (12 stimulations per day). LFS (0.1 ms pulse duration at 1 Hz, 200 pulses, and 50-150 muA) was applied to the perforant path immediately after termination of each rapid kindling stimulation. 1,3-Dimethyl-8-cyclopenthylxanthine (CPT; 50 muM), a selective A(1) antagonist and ZM241385 (ZM, 200 muM), a selective A(2A) antagonist were daily microinjected into the lateral ventricle 5 min before kindling stimulations. LFS had an inhibitory effect on kindling development. Pretreatment of animals with CPT reduced the inhibitory effect of LFS on kindling rate and suppressed the effects of LFS on potentiation of population EPSP during kindling acquisition. In addition, CPT was able to antagonize the effects of LFS on kindling-induced increase in early (10-50 ms intervals) and late (300-1000 ms intervals) paired pulse depression. ZM pretreatment had no effect on antiepileptogenic effects of LFS in kindling acquisition. In addition, LFS prevented the kindling-induced elevation of cyclic AMP (cAMP) levels in kindled animals. Based on these results, we suggest that the antiepileptogenic effects of LFS on perforant path kindling might be mediated through activation of adenosine A(1), but not A(2A) receptors. Moreover, modulation of cAMP levels by LFS may potentially be an important mechanism which explains the anticonvulsant effects of LFS in kindled seizures.

Rice blast disease, caused by the fungus Magnoporthe grisea is responsible for considerable damages on rice and leaf spot on some weeds in Iran and in other parts of the world. Infected samples were collected from rice and weeds including Digitaria sanguinalis (crabgrass), Setaria italica (foxtail millet), Echinochloa crus-galli (barnyard millet), and some unknown weeds during 1997-2005 and were preserved in collection of Mycology at the University of Tehran, Iran. In this study, genetic diversity of Magnaporthe grisea species complex isolates was studied based on DNA fingerprinting by rep-PCR, using of two primers including ERIC and BOX. The total DNA of 75 isolates was extracted and DNA fragments were amplified in a thermal cycler program using mentioned primers. Therefore, DNA fragments from 400 bp to 3000 bp were amplified. Based on cluster analysis for two primers (ERIC and BOX), eight fingerprinting groups (ctonal lineages) and sixty haplotypes were identified. "A" clonal lineage was containing the highest number of isolates and became dominant clonal lineages with 35 isolates from rice and 3 isolates from S. italica, whereas the highest number of isolates obtained from D. sanguinalis belonged to "E" clonal lineage and was the second largest clonal lineage. Approximately all of the M. grisea species complex isolates from crabgrass and some of unknown weeds were separated from other isolates in 42% similarity. As a result, asexual fertility causes low diversity in populations of M. grisea species complex and speciation could be one of the reasons of differentiation between isolates from D. sanguinalis with other isolates. Overall, these data indicated a low level of genetic diversity in the Iranian M. grisea species complex population similar to that reported in other countries.

Chronic pain has been reported to induce apoptosis. Both chronic excitation of neural pathways involved in pain transmission and control and the stress of pain may be potentially involved in apoptosis induced by pain. Here, we have investigated their possible role in pain-induced apoptosis. Inflammatory pain was induced by injection of formalin in intact and adrenalectomized (ADX) rats. Following exposure to repeated injections of 5% formalin, we detected Bax, Bcl-2, pro-caspase and activated caspase-3 proteins using immunoblotting. The results were compared with those obtained from animals suffered from chronic immobilization stress (IMO). These results showed an increased ratio of Bax/Bcl-2 and activated caspase-3 in hippocampus and dorsal lumbar spinal cord of animals treated with pain and IMO stress; these effects were reduced in ADX animals. On the other hand, the remaining apoptotic effect of pain in adrenalectomized rats was also significant. We surmise that both chronic neural activation and the stress induced by pain are involved in pain-induced apoptosis.

Irrigation with municipal effluent was evaluated during 25 months in Southern Iran from 2003 to 2005 in which 14 tree species were irrigated with effluent and borehole water at an annual supply rate of 3,940 and 5,395 m(3) ha(-1), respectively. To mitigate the environmental effects, a drip irrigation system was designed and the amount of applied water based on pan evaporation was measured by flow meters and soil properties were monitored. The statistical results showed that the applied effluent had no adverse effect on soil properties. The soil salinity was reduced from 8.2, 6.8 and 7.0 dSm(-1) to 1.07, 1.12 and 3.5 dSm(-1 )in the soil layers 0-30, 30-60 and 60-90 cm, respectively. The SAR decreased significantly, while soil pH increased by 0.8 and 0.6 units in the layers 0-30 and 30-60 cm. A total application of 9,335 m(3)ha(-1 )of effluent with a nitrogen and phosphorus concentration of 7.9 and 10.3 mg l(-1), added 73 and 101 kg ha(-1) of nitrogen and phosphorus to the soil. Organic carbon also increased significantly. Twenty-five months irrigation with effluent caused a slight increase in soil bulk density and a slight decrease in mean permeability. Because of an efficient filtration and high discharge rate of bubblers (drippers), no considerable sign of clogging was observed.

There are some reports indicating that adrenalectomy significantly potentiates morphine-induced analgesia. Since G-protein subunits have an important role in morphine effects at the cellular level and the exact mechanism(s) of adrenalectomy-induced morphine sensitization has not yet been clarified, the present study was designed to determine the changes in the levels of Galphai/o, Galphas, Gbeta mRNA involved in this phenomenon. All experiments were carried out on male Wistar rats. The tail-flick test was used to assess the nociceptive threshold and corticosterone levels were measured by radioimmunoassay as a marker of HPA function. The dorsal half of the lumbar spinal cord was assayed for the expression of G-protein subunits using semiquantitative PCR normalized to beta-actin gene expression. Results showed that morphine not only in 3 mg/kg, but also in a sub-effective dose (2 mg/kg) could affect the nociceptive threshold and induce an analgesic response in adrenalectomized (ADX) rats while 2 mg/kg morphine did not demonstrate analgesic properties in sham-operated animals. These effects were reversed with corticosterone replacement. Morphine increased plasma corticosterone concentration in a dose-dependent manner in sham-operated rats. Following adrenalectomy a significant increase in the mRNA levels of Galphai/o (79%) and Gbeta (96%) was observed in the dorsal portion of the lumbar spinal cord. In contrast, no significant changes were observed in the mRNA level of Galphas. In conclusion, our results demonstrate that the levels of the cellular components involved in morphine analgesia significantly increase in ADX animals. This may be at least partly responsible for adrenalectomy-induced morphine sensitization.

Opioids can induce analgesia and also hyperalgesia in humans and in animals. It has been shown that systemic administration of morphine induced a hyperalgesic response at an extremely low dose. However, the exact mechanism(s) underlying opioid-induced hyperalgesia has not yet been clarified. Here, we have investigated cellular events involved in low-dose morphine hyperalgesia in male Wistar rats. The data showed that morphine (0.01 microg i.t.) could elicit hyperalgesia as assessed by the tail-flick test. G(alphas) mRNA and protein levels increased significantly following exposure to the hyperalgesic dose of morphine. Furthermore, morphine at an analgesic dose (20 microg i.t.) significantly decreased cAMP levels in the dorsal half of the lumbar spinal cord, whereas the tissue cAMP levels were not affected by morphine treatment at a hyperalgesic dose. Intrathecal administration of nifedipine, an L-type calcium channel blocker, antagonized the hyperalgesia induced by the low-dose of morphine. Furthermore, pretreatment with the selective protein kinase C (PKC) inhibitor chelerytrine resulted in prevention of the morphine-induced hyperalgesia. KT 5720, a specific inhibitor of protein kinase A (PKA), did not show any effect on low-dose morphine-induced hyperalgesia. These results indicate a role for G(alphas), the PLC-PKC pathway, and L-type calcium channels in intrathecal morphine-induced hyperalgesia in rats. Activation of ordinary G(alphas) signaling through cAMP levels did not appear to play a major role in the induction of hyperalgesia by low-dose of morphine., ((c) 2007 Wiley-Liss, Inc.)

Activation of hypothalamic-pituitary-adrenal (HPA) axis inhibits development of morphine tolerance. Also, the expression of CaMKIIalpha is increased following chronic administration of morphine. In the current study, we tried to examine the effect of epinephrine, on the development of morphine tolerance; and also evaluate the expression of CaMKIIalpha as a molecular index for tolerance development. Analgesic tolerance was induced by intrathecal (i.t.) injection of morphine 15 microg/rat, twice a day for 5 days. To study the effect of epinephrine on development or reversal of morphine tolerance, epinephrine was administrated 20 min before morphine injections. Analgesia was assessed using tail flick test. Gene expression assays were done using RT-PCR. Following 5 days of combined administration of morphine and epinephrine (2, 5 or 10 microg/rat), in day 6, morphine produced potent analgesia. Administration of saline and morphine during days 1-5, caused reduced analgesic effect of morphine on day 6. After tolerance induction during 5 days, co-administration of epinephrine and morphine for another 5 days, significantly reversed the tolerance. Both morphine and epinephrine increased the expression of CaMKIIalpha. The expression of CaMKIIalpha was highly increased following combined administration of epinephrine and morphine. Our results showed the inhibition and reversal of analgesic tolerance to local administrated morphine by epinephrine. We observed the increased expression of CaMKIIalpha without development of morphine tolerance in animals treated with combined epinephrine and morphine.

Objective: This study aimed to compare the systemic bioavailability of two aciclovir tablets, Rouz-Aciclovir (test) and Zovirax (reference), in 12 healthy volunteers., Methods: In a crossover design, each subject received a single oral dose of aciclovir 400 mg followed by a 7-day washout period. Plasma concentrations of aciclovir were measured for up to 12 hours using a validated high-performance liquid chromatography method with a lower limit of quantification of 50 microg/L., Results: The mean values of maximum plasma concentration (C(max)), time to C(max) (t(max)), area under the plasma concentration-time curve from time 0 to 12 hours (AUC(12)) and from time 0 to infinity (AUC(infinity)), and plasma half-life following administration of the test product were 999.6 microg/L, 2.08 h, 4911.2 microg/L . h, 5417.7 microg/L . h and 3.08 h, respectively, and for the reference product 775.8 microg/L, 2.58 h, 3862.1 microg/L . h, 4295.4 microg/L . h and 3.14 h, respectively. The test/reference geometric ratio for C(max) (90% CI) was 1.30 (97.1, 174.8). The test/reference geometric ratios for AUC(12) (90% CI) and AUC(infinity) (90% CI) were 1.26 (99.7, 159.1) and 1.24 (98.9, 155.6), respectively. Therefore, the 90% CIs of C(max), AUC(12) and AUC(infinity) were not within the acceptable range of 80 and 125 suggested by the US FDA bioequivalence guideline., Conclusion: The results of the present study suggest that the aciclovir test product was not bioequivalent to the reference product. The exact reasons for this remain to be determined. However, we think the difference should be attributed to the difference in the type and amounts of ingredients used in the formulation that probably affect the contact time of aciclovir with the sites of absorption in the gut.

Low-frequency stimulation (LFS) has antiepileptogenic effects on kindled seizures. In the present study, the role of galanin receptors in the inhibitory effect of LFS on perforant path kindling acquisition was investigated in rats. Animals were kindled by perforant path stimulation in a rapid kindling manner (six stimulations per day). LFS (0.1 ms pulses at 1 Hz, 600 pulses, and 80-150 microA) was applied immediately after termination of each kindling stimulation. M35 (0.5 and 1.0 nM per site), a nonselective galanin receptor antagonist and M871 (1.0 microM per site), a selective galanin receptor type 2 (GalR2) antagonist, were daily microinjected into the dentate gyrus before starting the stimulation protocol. The expression of GalR2 in the dentate gyrus was also investigated using semi-quantitative RT-PCR. Application of LFS significantly retarded the kindling acquisition and delayed the expression of different kindled seizure stages. In addition, LFS significantly reduced the increment of daily afterdischarge duration during kindling development. Intra-dentate gyrus microinjection of both M35 and M871 significantly prevented the inhibitory effects of LFS on kindling parameters. During the focal kindled seizure stages (1-3) M871 had no significant effect. However, during generalized seizure stages (4 and 5), M871 had the same effect as M35. Semi-quantitative RT-PCR also showed that after kindling acquisition, the GalR2 mRNA level decreased in the dentate gyrus but application of LFS prevented this decrease. Obtained results show that activation of galanin receptors by endogenous galanin has a role in mediating the inhibitory effect of LFS on perforant path-kindled seizures. This role is exerted through GalR1 during focal- and through GalR2 during generalized-kindled seizures.

The immunomodulatory effects of exogenous opioids on induction of acquired immunity during microbial infection are now well known; however, our knowledge about the relationship between endogenous opioid response and microbial infections is rudimentary. Here, we report the effect of administration of Naloxone (NLX), an opioid receptor antagonist, on induction of acquired immunity during primary herpes simplex virus type 1 (HSV-1) infection. BALB/c mice received NLX, twice daily, 2 h before infection with HSV-1 until 7 days after infection. Cell-mediated immunity was assessed by evaluating lymphocyte proliferation, interferon-gamma (IFN-gamma) production, delayed type hypersensitivity (DTH) and mortality rate after acute HSV-1 challenge. The findings showed that a higher level of cell-mediated immunity was induced in the NLX-treated animals compared to the control group after induction of HSV-1 infection. However, the data indicate similar neutralizing antibody production in NLX-treated animals and control animals. This observation and further studies in this field may lead to the use of NLX as an adjuvant for designing microbial vaccines and adjunctive therapy of viral infections.

Ultra-low doses of morphine (UL-morphine) induce hyperalgesia, which is assumed to be mediated by stimulatory G proteins (G(alphas)) signaling pathway. G(alphas) pathway inhibition and chronic stress both attenuate development of tolerance to analgesic effect of morphine. This study evaluated the effect of chronic stress on UL-morphine-induced hyperalgesia to find out if chronic stress interacts with the G(alphas) signaling pathway. Repeated daily forced swim stress was applied to induce chronic stress. UL-morphine (1 microg/kg, intraperitoneal)-induced hyperalgesia was assessed using the tail-flick test on day 6, in male rats that during days 1-5 received different treatments of swim stress, dexamethasone, swim stress following adrenalectomy (ADX) or swim stress after sham operation. Chronic stress by itself induced hyperalgesia in control and sham-operated rats but inhibited UL-morphine-induced hyperalgesia. In ADX animals, chronic stress did not produce hyperalgesia and could not inhibit UL-morphine-induced hyperalgesia. Chronic dexamethasone produced hyperalgesia but did not change the UL-morphine-induced hyperalgesia. Inhibition of UL-morphine hyperalgesia by chronic stress suggests that chronic stress interacts with the G(alphas) signaling pathway, which is responsible for UL-morphine-induced hyperalgesia. The absence of this effect in the ADX-rats or after repetitive dexamethasone administration demonstrates that hypothalamic-pituitary-adrenal (HPA) axis activation is necessary for controlling UL-morphine-induced hyperalgesia. Finally, the interaction of stress with the G(alphas) signaling pathway may provide an explanation for the inhibitory effect of stress on development of tolerance to the analgesic effect of morphine.

It has been shown that systemic administration of morphine induced a hyperalgesic response at an extremely low dose. We have examined the effect of nifedipine, as a calcium channel blocker, on morphine-induced hyperalgesia in intact and adrenalectomized rats and on hypothalamic-pituitary-adrenal axis activity induced by ultra-low dose of morphine. To determine the effect of nifedipine on hyperalgesic effect of morphine, nifedipine (2 mg/kg i.p. and 10 microg i.t.) that had no nociceptive effect, was injected concomitant with morphine (1 microg/kg i.p. and 0.01 microg i.t. respectively). The tail-flick test was used to assess the nociceptive threshold, before and 30, 60, 120, 180, 240 and 300 min after drug administration. The data showed that low dose morphine systemic administration could produce hyperalgesic effect in adrenalectomized rats equivalent to sham-operated animals while intrathecal injection of morphine only elicited hyperalgesia in sham-operated animals. Nifedipine could block morphine-induced hyperalgesia in sham and adrenalectomized rats and even a mild analgesic effect was observed in the adrenalectomized group which was reversed by corticosterone replacement. Systemic administration of low dose morphine produced significant increase in plasma level of corticosterone. Nifedipine has an inhibitory effect on morphine-induced corticosterone secretion. Thus, the data indicate that dihydropyridine calcium channels are involved in ultra-low dose morphine-induced hyperalgesia and that both the pattern of morphine hyperalgesia and the blockage of it by nifedipine are modulated by manipulation of the hypothalamic pituitary adrenal axis.

Low frequency stimulation (LFS) has an inhibitory effect on kindling acquisition. In the present study the effect of the perforant path LFS on induction of rapid perforant path kindled seizures and synaptic transmission in the dentate gyrus was investigated. Animals were kindled by perforant path stimulation in a rapid kindling manner (12 stimulations per day). In one group of animals LFS (0.1 ms pulse duration at 1 Hz, 200 pulses, and 50-150 microA) was applied to perforant path, immediately after termination of each rapid kindling stimulation. Application of LFS significantly retarded the kindling acquisition and increased the number of stimulations to achieved different kindled seizure stages. LFS also prevented an increment in the slope of field excitatory postsynaptic potentials and population spike amplitude during kindling. In addition, LFS significantly reduced the marked increase in early (10-50 ms intervals) and late (300-1000 ms intervals) paired-pulse depression induced by kindling. According to obtained results, it may be suggested that LFS of perforant path has a significant antiepileptogenic effect through inhibition of synaptic transmission in dentate gyrus. Meanwhile, LFS prevents an increase in the paired-pulse depression during kindling acquisition.

The Fertile Crescent represents the center of origin and earliest known place of domestication for many cereal crops. During the transition from wild grasses to domesticated cereals, many host-specialized pathogen species are thought to have emerged. A sister population of the wheat-adapted pathogen Mycosphaerella graminicola was identified on wild grasses collected in northwest Iran. Isolates of this wild grass pathogen from 5 locations in Iran were compared with 123 M. graminicola isolates from the Middle East, Europe, and North America. DNA sequencing revealed a close phylogenetic relationship between the pathogen populations. To reconstruct the evolutionary history of M. graminicola, we sequenced 6 nuclear loci encompassing 464 polymorphic sites. Coalescence analyses indicated a relatively recent origin of M. graminicola, coinciding with the known domestication of wheat in the Fertile Crescent around 8,000-9,000 BC. The sympatric divergence of populations was accompanied by strong genetic differentiation. At the present time, no genetic exchange occurs between pathogen populations on wheat and wild grasses although we found evidence that gene flow may have occurred since genetic differentiation of the populations.

Fluorescent pseudomonads that produce antibiotic 2,4-diacetylphloroglocinol (2,4-DAPG) are important group of PGRP that inhibit a broad spectrum of plant pathogenic fungi. Studying on genetic diversity of 2,4-diacetylphloroglucinol-producing fluorescent pseudomonads has been shown with special importance. The first step to investigate the genetic diversity of these bacteria is detecting of the genes required for the biosynthesis of this antibiotic. The objectives of the current study were detection of phlD gene within fluorescent pseudomonads by a PCR-based assay, and comparison of phenotypic and genotypic characteristics of fluorescent pseudomonads with proven biocontrol potential against some soil-borne phytopathogenic fungi. We used a collection of 47 fluorescent Pseudomonas spp. some with known biological control activity against Macrophomina phaseolina, Rhizoctonia solani, Phytophthora nicotianae var. parasitica, Pythium sp. and Fusarium sp. in vitro and the potential to produce known secondary metabolites such as, siderophore, HCN and protease. The results indicated that 66, 40.42, 63.82,48.94 and 27.65% of strains revealed antagonistic activity against R. solani, M. phaseolina, Pythium sp., P. nicotianae and Fusarium sp., respectively. Rhizoctonia solani recognized as the most vulnerable fungus. Among 47 strains, 76.59, 97.87 and 17% of strains produced protease, siderophore and HCN, respectively. We could detect phlD gene in strains P-5, P-32, P-47. Strain CHA0 was used as positive control for the detection this gene. Overall, there was no obvious link between the existence of phlD gene and inhibition of fungal growth or production of the antifungal metabolites in vitro. But in some strains such as CHA0 and P-5, we saw a link between the existence of phlD and antifungal activities. Studying on detection and diversity of phlD provides a fundamental knowledge for developing a rapid genetic screening system to identify a potential biocontrol strains.

It is previously reported that the HPA axis plays role in the inhibitory effect of pain on tolerance development to analgesic effect of opioids. The present study was designed to investigate whether the chronic co-administration of dexamethasone as a glucocorticoid is also able to prevent or reverse analgesic tolerance to morphine and to compare the expression of G(alphai/o) and G(beta) subunits of G proteins in the context of chronic dexamethasone, development of morphine tolerance and their combination. Analgesic tolerance to morphine was induced by chronic intraperitoneally (i.p.) administration of morphine 20 mg/kg to male Wistar rats weighing 200-240 g within 4 consecutive days and analgesia was assessed using tail-flick test. Chronic dexamethasone was applied using 4 daily i.p. injections. Lumbar spinal tissues were assayed for the expression of G(alphai/o) and G(beta) proteins using "semiquantitative PCR" normalized to beta-actin gene expression. Results showed that chronic administration of dexamethasone could reduce and reverse the development of tolerance in rats that received chronic i.p. injections of morphine. Chronic administration of dexamethasone significantly increased the expression of G(alphai/o), while chronic administration of morphine did not change its expression. The expression of G(beta), however, was increased after the chronic administration of morphine, but did not change after the administration of chronic dexamethasone. None of these increases were observed when morphine and dexamethasone were co-administered. We conclude that the development of tolerance to analgesic effect of morphine could be prevented and reversed by dexamethasone co-administration. The increase in G(alphai/o) genes expression produced by chronic dexamethasone may facilitate the opioid signaling pathway and compensate for morphine-induced tolerance.

Purpose: This paper studied the effect of post-burn local hyperthermia on burn induced injury., Methods: A second-degree burn injury was induced on the right and left flanks of Balb/c mice. Thirty-two burn wounds were divided into four groups. Opioid receptor blocking was done for groups 3 and 4 by intra-peritoneal administration of Naloxone (NLX) 30 min before the thermal injury. Local hyperthermia (45 degrees C, 30 s) was applied only for the burn wounds of groups 2 and 4. Twenty-four hours after burn injury, the burned wounds were assessed for the level of iNOS (by immunohistochemistry) and the number of hair follicles (as an indicator of tissue injury)., Results: The wounds that received hyperthermia (group 2) had significantly more hair follicles (p < 0.001) compared to the control wounds (group 1). There was no significant difference between the number of hair follicles and acute inflammation of group 1 and group 3 (NLX + burn). Group 4 (NLX + burn + hyperthermia) had significantly fewer hair follicles compared to group 1 (p < 0.001), group 2 (p < 0.001) and group 3 (p < 0.001). The level of iNOS in groups 1, 3 and 4 was not significantly different but significantly more than group 2 (p < 0.001, p < 0.001 and p < 0.001, respectively)., Conclusions: The results showed that local hyperthermia after second degree burn decreased the tissue injury and iNOS expression. It is also concluded that endogenous opioid response may have a key role in the above mentioned effects of post-burn local hyperthermia.

Sensitive and selective determination of valproic acid in plasma by high-performance liquid chromatography (HPLC) is usually achieved with pre-column derivatization. In the present work, the derivatization is omitted due to using a simple but highly selective plasma extraction procedure and an optimized chromatographic condition. Valproic acid and the internal standard octanoic acid were extracted from plasma samples with n-hexane under acidic condition followed by back-extraction into diluted triethylamine. Chromatography was performed on a CN column (250 x 4.6 mm, 5 microm) under isocratic elution with acetonitrile-40 mM aqueous sodium dihydrogen phosphate (30:70, v/v), pH 3.5. Detection was made at 210 nm and analyses were run at a flow-rate of 1 ml/min. The method was specific and sensitive with a quantification limit of 1.25 microg/ml and a detection limit of 0.1 microg/ml in plasma. The mean absolute recovery for valproic acid using the present plasma extraction procedure was 75.8%. The intra- and inter-day coefficient of variation and percent error values of the assay method were all in acceptable range. Calibration curves were linear (r>0.999) from 1.25 to 320 microg/ml in plasma.

One hundred and eighteen isolates of Rhizoctonia solani were gathered from infected roots and hypocotyls of bean (Phaseolus vulgaris L.) grown in the fields of Tehran Province, Iran. Two isolates of the collected samples belonged to binucleate and 81 isolates to multinucleate of R. solani. The multinucleate isolates showed different anastomosis groups as AG-4 (subg. AG-4 HGI, AG-4HGII), AG-6 and AG-2. In greenhouse, pathogenicity tests carried out on bean cv. Naz in randomized design with 4 replications and each replication (pots) with 5 seeds of bean. Infection was done with seeds of wheat which were infected to the fungus with pasteurized soil. Results showed that the highest disease severity was caused by AG-4 (Rs21) isolates, whereas AG-4 (Rs74) isolates were weakly pathogenic with 90% and 21% infection, respectively. In this test the major pathogenic isolates belonged to AG-4 and they caused seed rot and damping-off of bean and AG-6 isolates were non-pathogenic. Five isolates of the fungus with major pathogenicity (Rs7, Rs18, Rs21, Rs62 and Rs71) selected and used for the reaction with different cultivars of bean. In this test, the cultivars and lines of bean (Pinto, red, white, green) studied in factorial experiment as randomized block design with 4 replications (pots). Results showed that none of the cultivars was completely resistant, however green bean cv. Sanry and pinto cv. Shad with number 4.8 disease severities had the highest susceptibility to seed rot and damping-off and red bean cv. Goli with 2.58 had the lowest susceptibility to the infection. Reaction of the cultivars and lines to the isolates of R. solani was significantly different at 1% level. Isolates of the fungus, Rs7, Rs21 with 84%, 90% pathogenicity was more virulent than the others.

The possibility of introducing eye-wiping test as a model of acute pain was examined in rat, and it was compared with the well-known hot plate test. One drop of NaCl 5 M was placed into the animal eye, and the number of eye wipes with the ipsilateral forelimb was counted during 30 s. The withdrawal latency in hot plate test was also examined. Afterward, animals were treated with morphine (1, 2, 4, 6, 8 or 10 mg/kg), imipramine (25 mg/kg), sodium salicylate (250 mg/kg) or saline (i.p). After 30 min, the animals were tested again with eye-wiping and hot plate tests. Our results showed that morphine injection dose dependently decreased the number of eye wipes and increased the response latency to hot plate tests. There was a good correlation between the analgesic effects of morphine on responses to both tests, however, morphine produced more pain relief in eye-wiping test. Imipramine significantly decreased the number of eye wipes and increased the response latency to hot plate test, while sodium salicylate and saline injection did not. It may be concluded that the eye-wiping test can be used as a reliable method in trigeminal pain studies, which is sensitive to opioid and tricyclic antidepressant in rat.

There are some reports regarding the inhibitory effect of pain on tolerance development to analgesic effect of opioids. The present study was designed to investigate whether the chronic formalin induced pain is able to reverse analgesic tolerance to morphine and to evaluate the expression of G(alpha i/o) and G(beta) subunits of G proteins in the context of chronic pain, development of morphine tolerance and their combination. Morphine tolerance was induced by chronic systemic (intraperitoneally, i.p.) or spinal (intrathecally, i.t.) administration of morphine to male Wistar rats weighing 200-240 g and analgesia was assessed using tail flick test. Chronic pain was induced by 4 daily intraplantar injections of 50 microl of 5% formalin. Lumbar spinal tissues were assayed for the expression of G(alpha i/o) and G(beta) proteins using "semiquantitative PCR" normalized to beta-actin gene expression. Results showed that chronic formalin induced pain could reduce and reverse the development of tolerance in rats that had received chronic (i.p. or i.t.) administration of morphine. Chronic administration of morphine did not change G(alpha i/o) gene expression, while chronic pain significantly increased its expression. The expression of G(beta), however, was increased after the chronic administration of morphine, but did not change after the induction of chronic pain. None of these increases were observed when morphine and formalin were administered at the same time. Due to synchronous development of morphine tolerance and changes in expression of G(beta), it may be concluded that the development of tolerance to analgesic effect of morphine is partially mediated by increase in G(beta) gene expression. The increase in G(alpha i/o) genes expression produced by chronic pain may facilitate the opioid signaling pathway and compensate for morphine-induced tolerance.

It has been shown that nifedipine, as a calcium channel blocker, can attenuate the development of tolerance to the antinociceptive effect of morphine; however, the role of HPA axis on this action has not been elucidated. We examined the effect of nifedipine on morphine analgesic tolerance in intact and adrenalectomized (ADX) rats and on HPA activity induced by morphine. Adult male rats were rendered tolerant to morphine by daily injection of morphine (15 mg/kg i.p.) for 8 days. To determine the effect of nifedipine on the development of morphine tolerance, nifedipine (1, 2 and 5 mg/kg i.p.) was injected concomitant with morphine. The tail-flick test was used to assess the nociceptive threshold, before and 30 min after morphine administration in days 1, 3, 5 and 8. Our results showed that despite the demonstration of tolerance in both ADX and sham operated rats, nifedipine in ADX rats prevented morphine tolerance development at a lower dose (2 mg/kg) than in sham operated rats, however corticosterone replacement prevented nifedipine effect in ADX rats. Acute administration of morphine produced significant increase in plasma corticosterone level, and with repeated injection, a tolerance to this neurosecretory effect was developed. Nifedipine (5 mg/kg) attenuated the acute effect of morphine, but could not block its neurosecretory tolerance.

Verticillium wilt caused by Verticillium dahliae is a serious problem of olive trees leading to significant reduction in yield. Verticillium wilt of olive trees was first recorded in Iran 1996 and confirm as due to Verticillium dahliae Kleb. 101 isolates of V. dahliae from olive trees at deferent locations in north provinces of Iran were assigned to vegetative compatibility groups (VCGS), using nitrate non-utilizing (Nit) mutants. A higher frequency of nit 1/nit 3 mutants (93%) was obtained compared with NitM (7%) with 10% of the isolates being assigned to VCG1 and 51% VCG4B and 19% VCG2A. 20% of isolates could not be classified in standard isolates. The pathogenecity of 15 randomly selected isolates (5 of each VCG) was tested on olive seedling (cv. Zard) and eggplant. The VCGs isolates were similarly aggressive on olive. However, VCG1 isolates were more aggressive on eggplant cv. Local than the VCG2A and VCG4B isolates as indicated by a higher colonization index. The pathogenecity tests of the pathogen on test plants (cotton cv. 'sahel', eggplant cv. 'local' and tomato cv. 'ps') show all isolates category in 2 pathogenecity groups defoliate and non-defoliate (with severe and mild subgroups). The morphology of V. dahliae isolates on C'zapeck's agar and water agar medium were different especially for microsclerotia appearance time in culture and their morphology.

Sugar beet leaves covered by sexual (cleistothecia) and asexual forms (mycelia and conidia) of Erysiphe betae were gathered at harvest time and maintained under natural outdoor conditions. In order to determine the function of cleistothecia and also conidia in the overwintering of E. betae some experiments were performed. The results showed that ascospores were unable to be released in petri dishes but their release under natural conditions occurred after 4 months. Under In vitro conditions ascospores did not germinate but on the leaves germination was rarely possible, however these ascospores were degraded after 7 days and didn't have pathogenicity. Conidia could induce pathogenicity after 3 but not 4 months. The period after inoculation until appearance of disease symptoms increased with aging of conidia. The results for conidial germination showed that fresh conidia had 80 percent germination on glass slides but it decreased sharply after 2 weeks and reached to 0 percent after 4 weeks. Although their germination on the leaves was not more than 46 percent of fresh conidia but they had good germination after 2 and 4 weeks. The results for the experiment to observe the first appearance of the disease in the field suggested that the first conidia were trapped by spore-trap in early June and the first symptoms appeared 20 days later. The conclusive results showed that ascospores had no function in the survival of the fungus and air-borne conidia are the main source of primary infections.