The relative gene transfer was calculated by dividing the % value

The relative gene transfer was calculated by dividing the % value of each treatment by the % value for the standard. Here transconjugants serve as standard. Data were analyzed using Graph Pad InStat-3 and expressed as mean ± standard

deviation (SD) of three independent experiment. The continuous variables were tested with one-way analysis of variance (ANOVA) and Dunnett’s test. Values < 0.05 was considered statistically significant. Re-identification of all of the clinical isolates were done and found to be of A. baumannii, C. braakii, E. coli, P. aeruginosa and K. pneumoniae. A. baumannii and C. braakii were positive for both qnrA and qnrB gene, whereas E. coli, P. aeruginosa and K. pneumoniae were positive for qnrB gene and none of the clinical isolates harbored qnrS ( Fig. 1). As shown in the Table 1, Potentox emerged as the most active antibacterial against A. baumannii, P. aeruginosa, E. coli and K. pneumoniae with MIC values 8 μg/ml. Epigenetic inhibitor solubility dmso The corresponding MIC for C. braakii was 16 μg/ml. The imipenem MIC values for A. baumannii and K. pneumoniae were 256 μg/ml each; 64 μg/ml for P. aeruginosa and C. braakii and 32 μg/ml for E. coli. The meropenem MIC values for A. baumannii, and K. pneumoniae were 128 μg/ml

each and 32 μg/ml for C. braakii and P. aeruginosa whereas 16 μg/ml for E. coli. For the other comparator drugs, the overall MIC values ranged from 32 to 1024 μg/ml. On the other hands, P. aeruginosa and K. pneumoniae found to be resistant to cefoperazone + sulbactam, amoxicillin plus clavulanic acid and levofloxacin; A. baumannii also showed resistant to amoxicillin plus clavulanic this website acid. There was a significant (p < 0.01) reduction in the MIC values of Potentox when compared

with the other comparator antibacterial agents ( Table 2). The zones of inhibition were calculated in millimeter for all strains and presented in the Table 3. Potentox was found to be sensitive against all clinical isolates as evident by zone of inhibition values, 23.5 ± 1.2, 20.8 ± 2.8, 25.8 ± 3.0, 27.2 ± 2.8, 23.2 ± 2.5 for A. baumannii, C. braakii, P. aeruginosa, E. coli and K. pneumoniae, respectively. Imipenem was found to be sensitive only against E. coli, aminophylline whereas meropenem was sensitive against P. aeruginosa and E. coli. Piperacillin plus tazobactam and cefoperazone plus sulbactam exhibited sensitivity toward C. braakii and E. coli. Cefepime was found to be sensitive only against C. braakii. Other tested drugs including amoxicillin plus clavulanic acid, moxifloxacin, levofloxacin and amikacin were observed to be resistant against all of the clinical isolates. The statistical analysis of AST values of Potentox vs other comparator drugs are shown in Table 4. Following conjugation, transconjugants were selected on MacConkey agar plates containing sodium azide and streptomycin. Analysis of transconjugants through PCR confirmed that transconjugants carrying the same gene as donor (Fig. 2).

They recorded neuronal responses to white noise, short bars, and

They recorded neuronal responses to white noise, short bars, and natural images. RF models selleck kinase inhibitor generated from each were tested for predictive accuracy with matching-type and cross-type stimuli. White noise stimuli elicited weak neural responses, resulting in noisy models, whereas bars and natural images elicited stronger responses and more accurate models. Natural image based models performed

better in cross-type validation than models from the two artificial stimuli, again suggesting that artificial stimuli may be poor probes for RF mapping. Tan and Yao examined the power spectra of natural scenes, and found that LGN neurons have spatio-temporal frequency tuning that acts as an optimal linear filter to maximize information transmission of natural scenes (Tan and Yao, 2009). They found that the power spectra vary significantly across different scenes and speculated that the spatio-temporal frequency characteristics of LGN neurons may be tuned to the frequencies of largest variability in natural scene spectra in order to assist in discrimination of natural stimuli. Mante et al. proposed

a model which, using the same parameters that apply to simple stimuli, predicts most of the firing rate responses to complex stimuli like natural scenes (Mante et al., 2008), including an important role for ECRF suppression in contrast gain control. They combined a standard center-surround CRF with fast-adapting gain control factors driven by local luminance and local contrast in the ECRF, and found excellent BVD-523 nmr predictive power for the model, except for bursting. For further information on the topic of natural scenes, we refer the reader to Simoncelli and Olshausen until (2001) review on the statistical methods available to analyze natural scene responses. They present an in-depth discussion of the efficient coding hypothesis and its applications, including single and

multiple neuron encoding. Simoncelli also offers a concise review of natural scene statistics (Simoncelli, 2003), including more efficient coding hypothesis discussion that includes some criticisms of the method and proposals of how to experimentally test its validity. Much of the early work in RF mapping used drifting bars or gratings with analysis techniques such as static maps created by line-weighting functions (Baker and Cynader, 1986 and Field and Tolhurst, 1986) and response-plane maps (Palmer and Davis, 1981 and Stevens and Gerstein, 1976). More recently the techniques of reverse correlation (Ringach and Shapley, 2004) driven by white noise (Chichilnisky, 2001) or M-sequence (Reid et al., 1997 and Sutter, 1991) visual stimuli to map and analyze receptive fields have been developed. A typical mapping paradigm is shown in Fig.

All other chemicals and reagents used in the study were of analyt

All other chemicals and reagents used in the study were of analytical grade. Matrix tablets of LAMI were prepared using various proportions of HPMC and a combination of HPMC and PEO as drug retarding polymers employing direct compression method. The drug, polymer(s) and all other excipients were sifted through 425 μm sieve (ASTM mesh no 40) and mixed uniformly. The dry blend was then blended with Aerosil and talc followed by magnesium stearate. The lubricated powder blends were characterized for drug content. The lubricated powder

blends were directly compressed on 16-station tablet compression machine (Cadmach Machinery Co, Ahmedabad, India) using 9 mm flat faced round (FFR) punches. Three batches were prepared for each formulation and compressed into 200 tablets from every batch for the characterization study. The drug content of the prepared matrix tablets

PLX4032 was determined in triplicate. For each batch, 20 tablets were taken, weighed and finely powdered. An CP-868596 order accurately weighed 300 mg of this powder was transferred to a 100 ml of pH 7.0 phosphate buffer, mixed for 10 min under sonication (Power sonic 505, HWASHIN Technology Co., Korea) and filtered through 0.45 μ (Millipore, India) filter. The sample was analysed after making appropriate dilutions using a UV spectrophotometer (UV-1700 E 23, Schimadzu, Japan) at 271.5 nm against blank.24 The weight variation was determined by taking 20 tablets using an isothipendyl electronic balance (ER182A, Mettler Toledo, Switzerland). Tablet hardness was determined for 10 tablets using a Monsanto tablet hardness tester (MHT-20, Campbell Electronics, Mumbai, India). Friability was determined by testing 10 tablets in a friability tester (FTA-20, Campbell Electronics, Mumbai, India) for 300 revolutions at 25 rpm. Moisture uptake studies on the powder blends and tablets was carried out at room temperature (30 ± 5 °C) and various relative humidity (RH) conditions such as 33%, 54% and 90% RH for assessing the varying environmental conditions during the manufacture process and storage.25 The

humid conditions of 33%, 54% and 90% RH were maintained by employing the saturated solutions of magnesium chloride, sodium dichromate potassium nitrate respectively. These solutions were transferred separately into three desiccators and allowed them for 24 h for saturation inside the chamber. Then accurately weighed powder blends and all the prepared tablets formulations were spread on petri plates and placed in each desiccator. The samples were weighed at 24, 48, 72, 96 and 120 h and the percent moisture uptake was determined. The in vitro dissolution studies were performed up to 14 h using the USP type I dissolution apparatus (Disso-2000, Labindia, Mumbai, India) at 100 rpm. The dissolution medium consisted of 900 ml of pH 7.0 phosphate buffer maintained at 37 ± °C as developed by Hwisa et al.

However, the mean percentage of CD8+ T-cells in group 4 was also

However, the mean percentage of CD8+ T-cells in group 4 was also significantly higher than in group 1, which showed a significantly higher CD4/CD8 rate as compared to all other groups. During previous DNA vaccination studies in SPF turkeys, unformulated pcDNA1/MOMP induced significant protection against severe clinical signs and lesions, bacterial replication and excretion following an experimental Cp. psittaci infection check details [24], [25],

[26] and [27]. However, complete protection was never observed. One might consider whether it will ever be possible to reach complete protection, if really needed at all. Maybe the previously used DNA vaccine could already create significant economical benefits by reducing the infection pressure and bacterial spread on the farms and as such diminishing Cp. psittaci outbreaks. Nevertheless, the potency of the previously used DNA vaccine can be further improved by optimising the efficiency of plasmid transfection and ompA translation inside host cells. We therefore tried to improve the immunogenicity of the DNA vaccine by optimising the ompA sequence selleck products for avian expression. Codon optimisation of ompA was performed

by Genscript corporation, increasing the codon adaptation index (CAI) [16] from 0.606 to 0.948. The codon-optimised ompA sequence was constructed synthetically, genetically linked to EGFP and cloned into pcDNA1, resulting in pcDNA1/MOMPopt. Subsequently, we tried to increase the transfection efficiency of the vaccine by generating pcDNA1/MOMPopt complexes using lPEI, brPEI, DOTAP/DOPE liposomes and starburst PAMAM dendrimers. else Non-cytotoxic complexes of pcDNA1/MOMPopt with liposomes, lPEI or brPEI significantly enhanced the transfection and translation efficiency in vitro compared to pcDNA1/MOMP, while complexes generated with dendrimers gave poor transfection results. Overall, the highest transfection efficiencies were obtained when using lPEI and brPEI complexes at an N/P ratio of 8. Administration of a Cp. psittaci vaccine

to poultry should be cost effective and easy. Aerosol administration could provide a solution, as most vaccines for avian respiratory diseases (New Castle Disease, Infectious Bronchitis or Avian Pneumovirus infections) are currently administered by aerosol or spray. Additionally, it has already been demonstrated that lPEI and brPEI are suitable gene delivery systems for aerosol therapy both in vitro and in mice [5], [6], [28], [29] and [30]. Stability of pcDNA1/MOMPopt lPEI and brPEI polyplexes and DNA integrity during nebulisation with a Cirrus™ nebulizer (Intersurgical) was therefore assessed by measuring particle size, zeta potential and DNA concentration in addition to agarose gel electrophoresis and expression in BGM cells.

Newer 1,2,3 – benzotriazole derivatives were synthesized by follo

Newer 1,2,3 – benzotriazole derivatives were synthesized by following green procedure under ultrasonic and solvent free conditions and characterized by spectral studies. All

the synthesized compounds were tested for anthelmintic activity against adult earthworms (P. posthuma) due to their anatomical and physiological resemblance with the intestinal roundworm parasites of human beings. 21 and 22 Albendazole, one of the reference compound in the present study is effective in a broad range of helminth infections, including round worms, hookworms, whipworms, pinworms and its mechanism of action involves inhibition of the glucose uptake system leading to a lethal depletion of energy reserves in the helminthes. 23 Another reference compound, mebendazole binds to the worm’s microtubular-protein ‘β-tubulin’ and inhibits its polymerization by blocking glucose this website C59 wnt uptake in the parasite 24, 25, 26 and 27 and also exhibits potent antitumor property both in vitro and in vivo. 28 From the observations made in the present study, higher concentration of the synthesized derivatives exhibited

paralytic effect much earlier and the time to death was shorter for worms. Out of the sixteen synthesized derivatives, four compounds (5B, 5F, 5J and 5N) showed anthelmintic activity in dose-dependent manner giving shortest time of paralysis (P) and death (D) with all three concentrations of the derivatives. These four compounds contain p-nitrophenyl substituent attached to azo group of benzotriazole Megestrol Acetate moieties and hence, displayed equal or comparable anthelmintic activity with reference to albendazole. Even earlier, best anthelmintic activity was reported for p-nitrophenyl substituted benzotriazoles like N1–(p-nitrophenyl) aminomethylene benzotriazole by Pawar. 29 Among these four derivatives (5B, 5F, 5J and 5N), 5J showed superior activity which might be due to attachment of additional p-nitrophenyl substituent to the

cyano group. Though, mebendazole was found to be effective compared to albendazole against the selected worms for the present study (Table 2), for mass treatment of multiple infections with Ascaris, hookworm, and Trichuris, albendazole was the preferred benzimidazole derivative from the comparative efficacy study of albendazole and mebendazole carried out in Pattani Province–Thailand by Jongsuksuntigul.30 As the four synthesized compounds showed comparable anthelmintic activity to albendazole, these compounds may also be tested for multiple infections. Better anthelmintic activity of the four compounds (5B, 5F, 5J and 5N) can be attributed to the p-nitrophenyl substituent attached to azo group of benzotriazole moieties. Superior activity of 5J might be due to attachment of additional p-nitrophenyl substituent to the cyano group. As the four synthesized compounds showed comparable anthelmintic activity to albendazole, these compounds may also be tested for multiple infections.

, 2011), thus suggesting that these mice exhibit a modest overact

, 2011), thus suggesting that these mice exhibit a modest overactivation of the HPA axis. On the other hand, it has been reported that ablation of adult hippocampal neurogenesis by X-ray irradiation does not impair basal HPA axis activity (Surget et al., 2011). Interestingly however, it has been reported that normalisation of HPA-axis overactivity by the antidepressant fluoxetine is dependent on intact adult hippocampal neurogenesis (Surget et al., 2011). Most studies report that ablating neurogenesis in rodents either with X-irradiation or with methylazoxymethanol (MAM) does

not increase their susceptibility to stress-induced changes in depression-related behaviour when compared with stressed neurogenesis-intact animals (Schloesser et al., 2010, Jayatissa et al., 2009, Lehmann et al., 2013 and Surget et al., 2011). For example, chemical ablation

of neurogenesis Ibrutinib chemical structure in rats with chronic injection of MAM did not see more induce anhedonia, a behaviour frequently observed following chronic stress, even though MAM reduced hippocampal cell proliferation to similar extent as exposure to a stress protocol (Jayatissa et al., 2009). Moreover, ablating neurogenesis prevented the ability of social defeat stress to induce social avoidance behaviour, thus suggesting that inhibiting neurogenesis may promote resilience rather than susceptibility to behavioural changes induced by this particular stressor (Lagace et al., 2010). Conversely, some studies have reported that neurogenesis ablated animals show a depressive-like

phenotype and increased susceptibility to stress-induced depression-like behaviour, including anhedonia and increased immobility in the forced swim test (Snyder et al., 2011 and Mateus-Pinheiro et al., 2013). Taken together, the precise role of adult hippocampal neurogenesis in stress susceptibility remains unclear as a lack of association as well as associations with both increased susceptibility and increased resilience being reported. On the other hand, it appears that adult hippocampal neurogenesis is required else for the ability of environmental factors such as environmental enrichment and physical exercise to promote stress resilience (Schloesser et al., 2010). Specifically, it was recently demonstrated that adult hippocampal neurogenesis is required for the beneficial effects of an enriched environment on antidepressant-like behaviours and recovery from stress-induced changes in behaviour (Schloesser et al., 2010). Stressed animals, when housed in an enriched environment, are rescued of the typical submissive behaviour induced by psychosocial stress, while animals housed in impoverished environment present a susceptible behaviour (Schloesser et al., 2010).

To assess the effects of CHO10 on cell proliferation, HER2-positi

To assess the effects of CHO10 on cell proliferation, HER2-positive and -negative cells were treated with different concentrations of CHO10 for 48 h. The growth of the tested cell lines was inhibited in a dose-dependent manner. In particular, CHO10-mediated growth inhibition was more potent in the AU-565, BT474 and SK-BR-3 cell lines, which are all HER2-overexpressing breast cancer cells (Cho et al., 2010 and Chrestensen et al., 2007). The growth inhibition caused by a 5 μM treatment of CHO10 was 88.6% in AU-565, 87.7% in BT474 and 87.1% in SK-BR-3; the growth inhibition of CHO10 was 65.0% in MCF-7, which is a breast cancer cell line that expresses a basal level of HER2,

Angiogenesis inhibitor and 40.2% in HEK293, which is a HER2-negative human embryonic kidney cells (Fig. 2A). Overall, these results suggest that CHO10 preferentially suppresses the growth of HER2-overexpressing

cancer cells. The percentage of apoptotic cells in the sub G1 peak of compound-treated SK-BR-3 was analyzed by FACS. As displayed in Fig. 2B, after the SK-BR-3 cells were check details treated with 10 μM of each compound for 24 h, a greater number of CHO10-treated cells (48.1%) started to undergo apoptosis than those treated with CHO3 (29.8%) or canertinib (30.8%). CHO10 induced apoptosis in the SK-BR-3 cells in a dose- and time-dependent manner, which was detected by observing the increase of the sub G1 peak in Fig. 2C and D. Cleaved PARP was used as a marker for apoptosis and was measured by western blotting.

CHO10 induced the corresponding increase of the PARP cleavage more potently than CHO3 but less potently than canertinib (Fig. 2E). Caspase-3 cleavage was not detected in the SK-BR-3 cells when they were treated with 10 μM CHO10 (Fig. 3A) for up to 8 h, even though CHO10-induced PARP cleavage was observed (Fig. 2E). To confirm this observation, the viability of SK-BR-3 cells was measured after treatment with CHO10 at concentrations of 0, 1, 5, 10, 15 and 25 μM in the absence and presence of 2 μM z-VAD-FMK for 48 h. The CHO10-induced cell death was not prevented by the use of the broad-spectrum caspase inhibitor z-VAD-FMK, as shown in Fig. 3B. The combination of CHO10 Histone demethylase and TAM exhibited greater inhibition of cell proliferation than TAM alone or the combination of TAM and canertinib (Fig. 4) in BT474 cells. The breast cancer cell line BT474 comprises ER-positive breast cancer cells and expresses high levels of amplified in breast cancer I (AIB1) and HER2. Because of these characteristics, BT474 is a perfect experimental model for TAM-resistance in ER-positive breast cancer cells (Su et al., 2008). Co-treatment of BT474 cells with CHO10 (1 μM) and TAM inhibited cell growth more strongly than TAM alone, accounting for 16.1% to 84.3% growth inhibition when treated with 5 μM of TAM for 72 h.

The crude extracts were evaporated to dryness using rotary evapor

The crude extracts were evaporated to dryness using rotary evaporator. The phytopathogenic fungi P. aphanidermatum was procured from Horticultural Research Station, Ambajipeta,

Andhra Pradesh. R. solani (MTCC 4633), P. oryzae (MTCC 1477), Curvularia oryzae (MTCC 2605) and F. oxysporum (MTCC 287), were procured from Microbial Type Culture Collection (MTCC), IMTECH, Chandigarh were used as test organisms. The strains were maintained and Vemurafenib mw tested on Potato Dextrose Agar (PDA). Antifungal activity of crude extracts of leaves of C. decandra Griff. Ding Hou was determined at concentrations of 100 μg/mL, 250 μg/mL, and 500 μg/mL by calculating zone of inhibition diameter (IZD) using Agar cup method. 12 and 13 Under aseptic conditions the PDA medium was poured into sterile petri plates and after the medium in the plates solidified, 1 × 108 spores ml−1 of fungal strains were inoculated and uniformly spread over the agar surface with a sterile L-shaped glass rod. Different concentrations of solutions were prepared by dissolving the crude compounds in Dimethyl Sulphoxide selleck kinase inhibitor (DMSO) and 100 μg/mL concentrations were prepared. After incubation, cups were scooped out with 6 mm sterile cork borer and the lids of the dishes were replaced. To each cup different concentrations of compounds

(100 μg/mL, 250 μg/mL, and 500 μg/mL) were added. All the plates were incubated at 28 °C, for 24 h and inhibition zones were observed and measured in mm. The average value of three replications was calculated for each experiment. Clotrimazole was used as positive control. Bioassays were conducted using laboratory reared 3rd and 4th instar S. litura (Fab.) larvae. Insects were reared on castor leaves (Ricinus communis) at room temperature (24–28 °C) under an L16:D8 photoperiod. Larvicidal activity (measured

mafosfamide as mortality after 24 h) of the crude extracts of C. decandra leaves was determined by topical application to early third and fourth instar larvae of S. litura according to Hummelbrunner et al. 14, 15, 16, 17, 18 and 19 Lethality was estimated by applying different concentrations (100–5000 μg/mL) of the crude extracts. Ten larvae as a set were tested per dose, in triplicate. A probit analysis was employed to calculate LD50 and LD90 concentrations. 20 The early 3rd and 4th instar stages laboratory-reared strains of A. aegypti were exposed to sublethal concentrations of 100–3000 μg/mL of the crude extracts by dissolving the extracts in acetone (99.8%) according to standard WHO procedure. 21 The larvae were fed with dry yeast powder by sprinkling on the water surface. The dead larvae were counted after 24 h and percentage mortality was reported from the average for the three replicates. A probit analysis using a computer program was employed on the results to determine LD50 and LD90 concentrations. The Gas chromatography–Mass spectrometry (GC–MS) analysis of methanol, chloroform and ethanol extracts of C.

We also determined the antibacterial activity of the extract agai

We also determined the antibacterial activity of the extract against Gram-positive and Gram-negative bacteria. All the solvents and chemicals used in this study were of analytical grade and obtained from HiMedia, Mumbai, India. 2,2-dipicryl-1-picrylhydrazyl (DPPH) was obtained from Sigma Chemical Co., St. Louis, MO, USA. The seeds of C. carvi were obtained from the supermarket located in Ontikoppal, Mysore, Karnataka, India. The C. carvi seeds were

cleaned, powdered and defatted using hexane in a Soxhlet apparatus for 6 h at 47 °C. The defatted C. carvi powder (10 g) was successively extracted with 100 ml water, 100 ml http://www.selleckchem.com/products/DAPT-GSI-IX.html 50% ethanol and 100 ml of equal mixture of 70% aqueous methanol and 70% aqueous acetone by stirring for 2 h at room temperature and the procedure was repeated Protein Tyrosine Kinase inhibitor thrice. All the respective extracts were combined and concentrated under vacuum in a rotary evaporator and subjected to hydrolysis with 2 N HCl to facilitate the breakage of glycosides. Further, the extract was phase separated with hexane

to remove any traces of fatty acids and subsequently with ethyl acetate (1:1) to extract polyphenolic compounds. The ethyl acetate phase was concentrated under vacuum and was kept at 4 °C until use. The total phenolic content of the extracts from three different solvent systems was estimated by Folin–Ciocalteau method.20 The phenolic content was expressed as gallic acid equivalents (GAE) of extract. The radical scavenging

activity of C. carvi phenolic extract was evaluated using DPPH as described earlier. 21 The changes in the absorbance of the samples were measured at 517 nm and the radical scavenging activity was expressed as the inhibition percentage using the following equation, %inhibition=[(O.D.ofblank−O.D.ofsample)/O.D.ofblank]×100 The samples were analyzed in triplicates and the IC50 value was calculated. The superoxide anion radicals were generated in a PMS-NADH system by the oxidation of NADH and assayed Sodium butyrate by the reduction of NBT.22 The scavenging activity was calculated using the equation %inhibition=[(O.D.ofblank−O.D.ofsample)/O.D.ofblank]×100 The samples were analyzed in triplicates and the IC50 value was calculated. The reducing power of C. carvi extract was determined according to the method of Oyaizu. 23 The average values of at least three measurements were plotted and compared with standards, BHA and BHT. The protective property of the C. carvi phenolic extract against oxidatively damaged DNA was determined using calf thymus DNA and analyzed by gel electrophoresis using 1% agarose/TAE buffer, at 60 V for 3 h. The DNA was visualized and photographed using a digital imaging system. The antibacterial activity of C. carvi phenolic extract was tested against food borne pathogens and food spoilage bacteria viz., Bacillus cereus, Escherichia coli, Staphylococcus aureus and Salmonella typhimurium by agar diffusion method with slight modifications.

They showed that the intravenous administration of Pyr and Oxa, w

They showed that the intravenous administration of Pyr and Oxa, which decreases blood Glu levels, accelerates the brain-to-blood Glu efflux. These results support the conclusion that the brain-to-blood Glu efflux can be modulated by changes in blood Glu levels

and can be accelerated by blood Glu scavenging (Gottlieb et al., 2003). Accordingly, Zlotnik and colleagues recently tested the effects of blood Glu scavengers in a rat model of closed head injury (CHI) and observed a significant improvement of the neurological recovery in the Oxa-treated and Pyr-treated rats when compared with saline-treated controls (Zlotnik et al., 2007 and Zlotnik et al., 2008). On these bases, we hypothesized that blood Glu scavenging induced by systemic Pyr and Oxa administration Selleckchem Talazoparib could be neuroprotective by increasing brain-to-blood

Glu efflux and thus preventing excitotoxic neuronal cell damage caused by prolonged epileptic seizures. In order to test this hypothesis, in the present HIF inhibitor investigation we studied the effect of Pyr and Oxa administration in rats subjected to pilocarpine-induced SE (Cavalheiro, 1995). Pilocarpine-induced SE is a widely used model to study neurodegeneration in limbic structures after prolonged epileptic seizures, particularly the hippocampal formation (Cavalheiro et al., 1991). Male Wistar rats (weight ∼250 g) were housed in groups of five under a continuous 12 h/12 h light/dark cycle and had free access to food and water. Experimental rats were injected with 4% pilocarpine hydrochloride (350 mg/kg i.p., Merck). Scopolamine methyl nitrate (1 mg/kg s.c., Sigma) was injected 30 min before pilocarpine to reduce the peripheral cholinergic effects. Approximately 10 min after pilocarpine

injection, animals developed partial limbic seizures with secondary generalization leading to self-sustained SE (Turski et al., 1983). After five hours, SE was blocked with diazepam (10 mg/kg i.p.). A control group received saline (-)-p-Bromotetramisole Oxalate instead of pilocarpine (Group Saline). Based on previous experiments designed to evaluate the neuroprotective effect of pyruvate and oxaloacetate in vivo (Lee et al., 2001, Gottlieb et al., 2003, Gonzales-Falcon et al., 2003 and Zlotnik et al., 2007), pyruvate solution (250 mg/kg, i.p., pH 7.4, Alfa Aesar) (Group Pilo + Pyr), oxaloacetate solution (1.4 mg/kg, i.p., pH 7.4, Calbiochem) (Group Pilo + Oxa) or both substances (Group Pilo + Pyr + Oxa) were administrated as single injection (1.5 ml) to rats thirty minutes after the development of SE. A control group received the same volume of saline instead of pyruvate and oxaloacetate (Group Pilo + Saline). Survival rates for each experimental group were calculated.