Search results for: inhibitory neurotransmitters
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 607

Search results for: inhibitory neurotransmitters

427 Supplementation of Citrulline with Lactic Acid Bacteria Protects Foodborne Pathogens Adhesion and Improves the Cell Integrity on the Intestinal Epithelial Cell

Authors: Sze Wing Ho, Nagendra P. Shah

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Lactic acid bacteria (LAB) have shown the beneficial effects on human gastrointestinal tract, such as protects diarrhea induced by lactose intolerance or enteric pathogens. Citrulline is a non-protein amino acid and also the precursors of arginine and nitric oxide, it has shown to enhance intestinal barrier function. Citrulline has shown to improve the growth of some strains of LAB, it is important for LAB to have a sufficient cell concentration to contribute the effects. Therefore, the aims of this study were to investigate the effect of combining citrulline with LAB on the anti-adhesion effect against pathogens and the effect on the cell integrity. The effect of citrulline on selected LAB was determined by incubating in 0%, 0.1% or 0.2% citrulline enriched MRS broth for 18 h. The adhesion ability of LAB and the anti-adhesion effect of LAB and citrulline against pathogens were performed on IPEC-J2 cell line. Transepithelial electrical resistance (TEER) assay was used to measure the tight junction (TJ) integrity. TJ proteins (claudin-1, occludin and zonula occluden-1 (ZO-1)) were determined by western blot analysis. It found that the growth of Lactobacillus helveticus ASCC 511 was significantly stimulated by 0.2% citrulline compared with control during 18 h fermentation. The adhesion of L. helveticus ASCC 511 and Lactobacillus delbrueckii ssp. bulgaricus (L. bulgaricus) ASCC 756 was increased when supplemented with citrulline. Citrulline has shown significant inhibitory effect on the adhesion of Escherichia coli PELI0480 (O157:H7), Shigella sonnei ATCC 25931, Staphyloccocus aureus CMCC26003 and Cronobacter sakazakii ATCC 29544. The anti-adhesion effect of L. helveticus ASCC 511, L. bulgaricus ASCC 756 and Lactobacillus paracasei ASCC 276 against Cronobacter sakazakii ATCC 29544 was significantly enhanced with citrulline supplementation. Treatments with citrulline and LAB were able to maintain the TEER of IPEC-J2 cell and shown the positive effect on the TJ proteins. In conclusion, citrulline had stimulating effect on some strains of LAB and determined to improve the adhesion of LAB on intestinal epithelial cell, to enhance the inhibitory effect on enteric pathogens adhesion as well as had beneficial effects on maintaining cell integrity. It implied LAB supplemented with citrulline might have advantageous effects on gastrointestinal tracts.

Keywords: citrulline, lactic acid bacteria, amino acid, anti-adhesion effect, cell integrity

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426 Study of Growth Behavior of Some Bacterial Fish Pathogens to Combined Selected Herbal Essential Oil

Authors: Ashkan Zargar, Ali Taheri Mirghaed, Zein Talal Barakat, Alireza Khosravi, Hamed Paknejad

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With the increase of bacterial resistance to the chemical antibiotics, replacing it with ecofriendly herbal materials and with no adverse effects in the host body is very important. Therefore, in this study, the effect of combined essential oil (Thymus vulgaris-Origanum magorana and Ziziphora clinopodioides) on the growth behavior of Yersinia ruckeri, Aeromonas hydrophila and Lactococcus garvieae was evaluated. The compositions of the herbal essential oils used in this study were determined by gas chromatography-mass spectrometry (GC-MS) while, the investigating of antimicrobial effects was conducted by the agar-disc diffusion method, determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and bacterial growth curves determination relied on optical density (OD) at 630 nm. The main compounds were thymol (40.60 %) and limonene (15.98 %) for Thymus vulgaris while carvacrol (57.86 %) and thymol (13.54 %) were the major compounds in Origanum magorana. As regards Ziziphora clinopodiodes, α-pinene (22.6 %) and carvacrol (21.1 %) represented the major constituents. Concerning Yersinia ruckeri, disc-diffusion results showed that t.O.z (50 % Origanum majorana) combined essential oil was presented the best inhibition zone (30.66 mm) but it was exhibited no significant differences with other tested commercial antibiotics except oxytetracycline (P <0/05). The inhibitory activity and the bactericidal effect of the t.O.z, unveiled by the MIC= 0.2 μL /mL and MBC= 1.6 μL /mL values, were clearly the best between all combined oils. The growth behaviour of Yersinia ruckeri was affected by this combined essential oil and changes in temperature and pH conditions affected herbal oil performance. As regard Aeromonas hydrophila, its results were so similar to Yersinia ruckeri results and t.O.z (50 % Origanum majorana) was the best between all combined oils (inhibition zone= 26 mm, MIC= 0.4 μL /mL and MBC= 3.2 μL /mL, combined essential oil was affected bacterial growth behavior). Also for Lactococcus garvieae, t.O.z (50 % Origanum majorana) was the best between all combined oils having the best inhibition zone= 20.66 mm, MIC= 0.8 μL /mL and MBC= 1.6 μL /mL and best effect on inhibiting bacterial growth. Combined herbal essential oils have a good and noticeable effect on the growth behavior of pathogenic bacteria in the laboratory, and by continuing research in the host, they may be a suitable alternative to control, prevent and treat diseases caused by these bacteria.

Keywords: bacterial pathogen, herbal medicine, growth behavior, fish

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425 Effect of a Mixture of Phenol, O-Cresol, P-Cresol, and M-Cresol on the Nitrifying Process in a Sequencing Batch Reactor

Authors: Adriana Sosa, Susana Rincon, Chérif Ben, Diana Cabañas, Juan E. Ruiz, Alejandro Zepeda

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The complex chemical composition (mixtures of ammonium and recalcitrant compounds) of the effluents from the chemical, pharmaceutical and petrochemical industries represents a challenge in their biological treatment. This treatment involves nitrification process that can suffer an inhibition due to the presence of aromatic compounds giving as a result the decrease of the process efficiency. The inhibitory effects on nitrification in the presence of aromatic compounds have already been studied; however a few studies have considered the presence of phenolic compounds in the form of mixtures, which is the form that they are present in real context. For this reason, we realized a kinetic study on the nitrifying process in the presence of different concentrations of a mixture of phenol, o-cresol, m-cresol and p-cresol (0 - 320 mg C/L) in a sequencing batch reactor (SBR). Firstly, the nitrifying process was evaluated in absence of the phenolic mixture (control 1) in a SBR with 2 L working volume and 176 mg/L of nitrogen of microbial protein. Total oxidation of initial ammonium (efficiency; ENH4+ of 100 %) to nitrate (nitrifying yield; YNO3- of 0.95) were obtained with specific rates of ammonium consumption (qN-NH4+) and nitrate production (qN-NO3-) (of 1.11 ± 0.04 h-1 and 0.67 h-1 ± 0.11 respectively. During the phase of acclimation with 40 mg C/L of the phenolic mixture, an inhibitory effect on the nitrifying process was observed, provoking a decrease in ENH4+ and YNO3- (11 and 54 % respectively) as well as in the specific rates (89 y 46 % respectively), being the ammonia oxidizing bacteria (BAO) the most affected. However, in the next cycles without the phenolic mixture (control 2), the nitrifying consortium was able to recover its nitrifying capacity (ENH4+ = 100% and YNO3-=0.98). Afterwards the SBR was fed with 10 mg C/L of the phenolic mixture, obtaining and ENH4+ of 100%, YNO3- and qN-NH4+ 0.62 ± 0.006 and 0.13 ± 0.004 respectively, while the qN-NO3- was 0.49 ± 0.007. Moreover, with the increase of the phenolic concentrations (10-160 mg C/L) and the number of cycles the nitrifying consortium was able to oxidize the ammonia with ENH4+ of 100 % and YNO3- close to 1. However a decrease in the values of the nitrification specific rates and increase in the oxidation in phenolic compounds (70 to 94%) were observed. Finally, in the presence of 320 mg C/L, the nitrifying consortium was able to simultaneously oxidize the ammonia (ENH4+= 100%) and the phenolic mixture (p-cresol>phenol>m-cresol>o-cresol) being the o-cresol the most recalcitrant compound. In all the experiments the use of a SBR allowed a respiratory adaptation of the consortium to oxidize the phenolic mixture achieving greater adaptation of the nitrite-oxidizing bacteria (NOB) than in the ammonia-oxidizing bacteria (AOB).

Keywords: cresol, inhibition, nitrification, phenol, sequencing batch reactor

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424 Inhibitory Effect of Hydroalcoholic Extract of Cestrum Nocturnum on α-Amylase Activity

Authors: Rajesh Kumar, Anil Kamboj

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Inhibition of α- amylase play a vital role in the clinical management of postprandial hyperglycemia. Although, powerful synthetic inhibitors are available, natural inhibitors are potentially safer. The present study was carried out to evaluate α- amylase inhibition activity from hydroalcoholic extracts from aerial parts of Cestrum nocturnum. Hydroalcoholic extract was prepared by Soxhletation Method. The extract showed strong inhibition towards α- amylase activity and IC50 value were 45.9 µg. This In vitro studies indicate the potential of C. nocturnum in the development of effective anti-diabetic agents.

Keywords: α- amylase, cestrum nocturnum, hyperglycemia, hydroalcoholic extracts, diabetes

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423 Antibacterial and Antifungal Activities of the Essential Oil of Pulicaria jaubertii Leaves

Authors: Methaq Algabr, Nabil Al-Hajj, Ameerh Jaber, Amtellah Alshotobi, Shaima'a Al-suryhi, Gadah Whaban, Nawal Alshehari

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Steam distillation of the essential oil of P. jaubertii was performed using a Clevenger apparatus. Essential oils were analyzed by gas chromatography-flame ionization detector (GC-FID) and gas chromatography coupled to chromatography–mass spectrometry (GC-MS). The major chemical components identified in P. jaubertii essential oil include carvotanacetone (63.975%), 1-methyl-1,2-propanedione (5.887%), 2,5-dimethoxy-para-cymene (3.303%) and ar-curcumene (3.276%). The antimicrobial activity of the essential oil of P. jaubertii was evaluated against all tested microorganisms. P. jaubertii essential oil inhibited all tested microorganisms except Escherichia coli with a minimum inhibitory concentration (MIC) of 5.0 μg/mL against Staphylococcus aureus.

Keywords: Pulicaria jaubertii, essential oil, antimicrobial, Carvotancetone

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422 In vitro Susceptibility of Isolated Shigella flexneri and Shigella dysenteriae to the Ethanolic Extracts of Trachyspermum ammi and Peganum harmala

Authors: Ibrahim Siddig Hamid, Ikram Mohamed Eltayeb

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Trachyspermum ammi belongs to the family Apiaceae, is used traditionally for the treatment of gastrointestinal ailments, lack of appetite and bronchial problems as well used as antiseptic, antimicrobial, antipyretic, febrifugal and in the treatment of typhoid fever. Peganum harmala belongs to the family Zygophyllaceae it has been reported to have an antibacterial activity and used to treat depression and recurring fevers. It also used to kill algae, bacteria, intestinal parasites and molds. In Sudan, the combination of two plants are traditionally used for the treatment of bacillary dysentery. Bacillary dysentery is caused by one or more types of Shigella species bacteria mainly Shigella dysenteri and shigella flexneri. Bacillary dysentery is mainly found in hot countries like Sudan with poor hygiene and sanitation. Bacillary dysentery causes sudden onset of high fever and chills, abdominal pain, cramps and bloating, urgency to pass stool, weight loss, and dehydration and if left untreated it can lead to serious complications including delirium, convulsions and coma. A serious infection like this can be fatal within 24 hours. The objective of this study is to investigate the in vitro susceptibility of Sh. flexneri and Sh. dysenteriae to the T. ammi and P. harmala. T. ammi and P. harmala were extracted by 96% ethanol using Soxhlet apparatus. The antimicrobial activity of the extracts was investigated according to the disc diffusion method. The discs were prepared by soaking sterilized filter paper discs in 20 microliter of serially diluted solutions of each plant extract with the concentrations (100, 50, 25, 12.5, 6.25mg/dl) then placing them on Muller Hinton Agar plates that were inoculated with bacterial suspension separately, the plates were incubated for 24 hours at 37c and the minimum inhibitory concentration of the extract which was the least concentration of the extract to inhibit fungal growth was determined. The results showed the high antimicrobial activity of T. ammi extract with an average diameter zone ranging from 18-20 mm and its minimum inhibitory concentration was found to be 25 mg/ml against the two shigella species. P. harmala extract was found to have slight antibacterial effect against the two bacteria. This result justified the Sudanese traditional use of Trachyspermum ammi plant for the treatment of bacillary dysentery.

Keywords: harmala, peganum, shigella, trachyspermum

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421 Inhibitory Action of Fatty Acid Salts against Cladosporium cladosporioides and Dermatophagoides farinae

Authors: Yui Okuno, Mariko Era, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Introduction: Fungus and mite are known as allergens that cause an allergic disease for example asthma bronchiale and allergic rhinitis. Cladosporium cladosporioides is one of the most often detected fungi in the indoor environment and causes pollution and deterioration. Dermatophagoides farinae is major mite allergens indoors. Therefore, the creation of antifungal agents with high safety and the antifungal effect is required. Fatty acid salts are known that have antibacterial activities. This report describes the effects of fatty acid salts against Cladosporium cladosporioides NBRC 30314 and Dermatophagoides farinae. Methods: Potassium salts of 9 fatty acids (C4:0, C6:0, C8:0, C10:0, C12:0, C14:0, C18:1, C18:2, C18:3) were prepared by mixing the fatty acid with the appropriate amount of KOH solution to a concentration of 175 mM and pH 10.5. The antifungal method, the spore suspension (3.0×104 spores/mL) was mixed with a sample of fatty acid potassium (final concentration of 175 mM). Samples were counted at 0, 10, 60, 180 min by plating (100 µL) on PDA. Fungal colonies were counted after incubation for 3 days at 30 °C. The MIC (minimum inhibitory concentration) against the fungi was determined by the two-fold dilution method. Each fatty acid salts were inoculated separately with 400 µL of C. cladosporioides at 3.0 × 104 spores/mL. The mixtures were incubated at the respective temperature for each organism for 10 min. The tubes were then contacted with the fungi incubated at 30 °C for 7 days and examined for growth of spores on PDA. The acaricidal method, twenty D. farinae adult females were used and each adult was covered completely with 2 µL fatty acid potassium for 1 min. The adults were then dried with filter paper. The filter paper was folded and fixed by two clips and kept at 25 °C and 64 % RH. Mortalities were determained 48 h after treatment under the microscope. D. farina was considered to be dead if appendages did not move when prodded with a pin. Results and Conclusions: The results show that C8K, C10K, C12K, C14K was effective to decrease survival rate (4 log unit) of the fatty acids potassium incubated time for 10 min against C. cladosporioides. C18:3K was effective to decrease 4 log unit of the fatty acids potassium incubated time for 60 min. Especially, C12K was the highest antifungal activity and the MIC of C12K was 0.7 mM. On the other hand, the fatty acids potassium showed no acaricidal effects against D. farinae. The activity of D. farinae was not adversely affected after 48 hours. These results indicate that C12K has high antifungal activity against C. cladosporioides and suggest the fatty acid potassium will be used as an antifungal agent.

Keywords: fatty acid salts, antifungal effects, acaricidal effects, Cladosporium cladosporioides, Dermatophagoides farinae

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420 Screening for Hit Identification against Mycobacterium abscessus

Authors: Jichan Jang

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Mycobacterium abscessus is a rapidly growing life-threatening mycobacterium with multiple drug-resistance mechanisms. In this study, we screened the library to identify active molecules targeting Mycobacterium abscessus using resazurin live/dead assays. In this screening assay, the Z-factor was 0.7, as an indication of the statistical confidence of the assay. A cut-off of 80% growth inhibition in the screening resulted in the identification of four different compounds at a single concentration (20 μM). Dose-response curves identified three different hit candidates, which generated good inhibitory curves. All hit candidates were expected to have different molecular targets. Thus, we found that compound X, identified, may be a promising candidate in the M. abscessus drug discovery pipeline.

Keywords: Mycobacterium abscessus, antibiotics, drug discovery, emerging Pathogen

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419 Stems of Prunus avium: An Unexplored By-product with Great Bioactive Potential

Authors: Luís R. Silva, Fábio Jesus, Catarina Bento, Ana C. Gonçalves

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Over the last few years, the traditional medicine has gained ground at nutritional and pharmacological level. The natural products and their derivatives have great importance in several drugs used in modern therapeutics. Plant-based systems continue to play an essential role in primary healthcare. Additionally, the utilization of their plant parts, such as leaves, stems and flowers as nutraceutical and pharmaceutical products, can add a high value in the natural products market, not just by the nutritional value due to the significant levels of phytochemicals, but also by to the high benefit for the producers and manufacturers business. Stems of Prunus avium L. are a byproduct resulting from the processing of cherry, and have been consumed over the years as infusions and decoctions due to its bioactive properties, being used as sedative, diuretic and draining, to relief of renal stones, edema and hypertension. In this work, we prepared a hydroethanolic and infusion extracts from stems of P. avium collected in Fundão Region (Portugal), and evaluate the phenolic profile by LC/DAD, antioxidant capacity, α-glucosidase inhibitory activity and protection of human erythrocytes against oxidative damage. The LC-DAD analysis allowed to the identification of 19 phenolic compounds, catechin and 3-O-caffolquinic acid were the main ones. In a general way, hydroethanolic extract proved to be more active than infusion. This extract had the best antioxidant activity against DPPH• (IC50=22.37 ± 0.28 µg/mL) and superoxide radical (IC50=13.93 ± 0.30 µg/mL). Furthermore, it was the most active concerning inhibition of hemoglobin oxidation (IC50=13.73 ± 0.67 µg/mL), hemolysis (IC50=1.49 ± 0.18 µg/mL) and lipid peroxidation (IC50=26.20 ± 0.38 µg/mL) on human erythrocytes. On the other hand, infusion revealed to be more efficient towards α-glucosidase inhibitory activity (IC50=3.18 ± 0.23 µg/mL) and against nitric oxide radical (IC50=99.99 ± 1.89 µg/mL). The Sweet cherry sector is very important in Fundão Region (Portugal), and taking profit from the great wastes produced during processing of the cherry to produce added-value products, such as food supplements cannot be ignored. Our results demonstrate that P. avium stems possesses remarkable antioxidant and free radical scavenging properties. It is therefore, suggest, that P. avium stems can be used as a natural antioxidant with high potential to prevent or slow the progress of human diseases mediated by oxidative stress.

Keywords: stems, Prunus avium, phenolic compounds, biological potential

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418 Anti-Acanthamoeba Activities of Fatty Acid Salts and Fatty Acids

Authors: Manami Masuda, Mariko Era, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Objectives: Fatty acid salts are a type of anionic surfactant and are produced from fatty acids and alkali. Moreover, fatty acid salts are known to have potent antibacterial activities. Acanthamoeba is ubiquitously distributed in the environment including sea water, fresh water, soil and even from the air. Although generally free-living, Acanthamoeba can be an opportunistic pathogen, which could cause a potentially blinding corneal infection known as Acanthamoeba keratitis. So, in this study, we evaluated the anti-amoeba activity of fatty acid salts and fatty acids to Acanthamoeba castellanii ATCC 30010. Materials and Methods: The antibacterial activity of 9 fatty acid salts (potassium butyrate (C4K), caproate (C6K), caprylate (C8K), caprate (C10K), laurate (C12K), myristate (C14K), oleate (C18:1K), linoleate (C18:2K), linolenate (C18:3K)) tested on cells of Acanthamoeba castellanii ATCC 30010. Fatty acid salts (concentration of 175 mM and pH 10.5) were prepared by mixing the fatty acid with the appropriate amount of KOH. The amoeba suspension mixed with KOH with a pH adjusted solution was used as the control. Fatty acids (concentration of 175 mM) were prepared by mixing the fatty acid with Tween 80 (20 %). The amoeba suspension mixed with Tween 80 (20 %) was used as the control. The anti-amoeba method, the amoeba suspension (3.0 × 104 cells/ml trophozoites) was mixed with the sample of fatty acid potassium (final concentration of 175 mM). Samples were incubated at 30°C, for 10 min, 60 min, and 180 min and then the viability of A. castellanii was evaluated using plankton counting chamber and trypan blue stainings. The minimum inhibitory concentration (MIC) against Acanthamoeba was determined using the two-fold dilution method. The MIC was defined as the minimal anti-amoeba concentration that inhibited visible amoeba growth following incubation (180 min). Results: C8K, C10K, and C12K were the anti-amoeba effect of 4 log-unit (99.99 % growth suppression of A. castellanii) incubated time for 180 min against A. castellanii at 175mM. After the amoeba, the suspension was mixed with C10K or C12K, destroying the cell membrane had been observed. Whereas, the pH adjusted control solution did not exhibit any effect even after 180 min of incubation with A. castellanii. Moreover, C6, C8, and C18:3 were the anti-amoeba effect of 4 log-unit incubated time for 60 min. C4 and C18:2 exhibited a 4-log reduction after 180 min incubation. Furthermore, the minimum inhibitory concentration (MIC) was determined. The MIC of C10K, C12K and C4 were 2.7 mM. These results indicate that C10K, C12K and C4 have high anti-amoeba activity against A. castellanii and suggest C10K, C12K and C4 have great potential for antimi-amoeba agents.

Keywords: Fatty acid salts, anti-amoeba activities, Acanthamoeba, fatty acids

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417 Geographic Variation in the Baseline Susceptibility of Helicoverpa armigera (Hubner) (Noctuidae: Lepidoptera) Field Populations to Bacillus thuringiensis Cry Toxins for Resistance Monitoring

Authors: Muhammad Arshad, M. Sufian, Muhammad D. Gogi, A. Aslam

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The transgenic cotton expressing Bacillus thuringiensis (Bt) provides an effective control of Helicoverpa armigera, a most damaging pest of the cotton crop. However, Bt cotton may not be the optimal solution owing to the selection pressure of Cry toxins. As Bt cotton express the insecticidal proteins throughout the growing seasons, there are the chances of resistance development in the target pests. A regular monitoring and surveillance of target pest’s baseline susceptibility to Bt Cry toxins is crucial for early detection of any resistance development. The present study was conducted to monitor the changes in the baseline susceptibility of the field population of H. armigera to Bt Cry1Ac toxin. The field-collected larval populations were maintained in the laboratory on artificial diet and F1 generation larvae were used for diet incorporated diagnostic studies. The LC₅₀ and MIC₅₀ were calculated to measure the level of resistance of population as a ratio over susceptible population. The monitoring results indicated a significant difference in the susceptibility (LC₅₀) of H. armigera for first, second, third and fourth instar larval populations sampled from different cotton growing areas over the study period 2016-17. The variations in susceptibility among the tested insects depended on the age of the insect and susceptibility decreased with the age of larvae. The overall results show that the average resistant ratio (RR) of all field-collected populations (FSD, SWL, MLT, BWP and DGK) exposed to Bt toxin Cry1Ac ranged from 3.381-fold to 7.381-fold for 1st instar, 2.370-fold to 3.739-fold for 2nd instar, 1.115-fold to 1.762-fold for 3rd instar and 1.141-fold to 2.504-fold for 4th instar, depicting maximum RR from MLT population, whereas minimum RR for FSD and SWL population. The results regarding moult inhibitory concentration of H. armigera larvae (1-4th instars) exposed to different concentrations of Bt Cry1Ac toxin indicated that among all field populations, overall Multan (MLT) and Bahawalpur (BWP) populations showed higher MIC₅₀ values as compared to Faisalabad (FSD) and Sahiwal (SWL), whereas DG Khan (DGK) population showed an intermediate moult inhibitory concentrations. This information is important for the development of more effective resistance monitoring programs. The development of Bt Cry toxins baseline susceptibility data before the widespread commercial release of transgenic Bt cotton cultivars in Pakistan is important for the development of more effective resistance monitoring programs to identify the resistant H. armigera populations.

Keywords: Bt cotton, baseline, Cry1Ac toxins, H. armigera

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416 Analysis of Expression of SP and NOS in the Porcine Nodose Ganglion (NG) Sensory Neurons Supplying Prepyloric Stomach Region after Intragastric Hydrochloric Acid Infusion

Authors: Liliana Rytel, Jarosław Całka

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One of the diseases that are very common health problem of modern man is the stomach hyperacidity. It is well known that this pathological state, during which gastric glands secrete too much of hydrochloric acid can be caused due to various factors such as stress, eating habits, alcohol, smoking and some, especially anti-inflammatory drugs. Moreover, hyperacidity is recognized as one of factors leading to development of peptic ulcer disease. Therefore, we analyzed expression of substance P (SP) and neuronal isoform of nitric oxide synthase (nNOS) in the porcine nodose ganglion sensory neurons innervating prepyloric stomach region in physiological state and following intragastric infusion of hydrochloric acid. The study was performed on 8 immature gilts of the Large White Polish breed. All animals were injected retrograde marker Fast Blue (FB) into the anterior prepyloric stomach wall. After injections of FB, pigs were divided into two groups: control (group C; n = 4) and experimental (HCL group, n = 4) and after convalescence period of 23 days, animals of HCL group were subjected to renewed anaesthesia. Then, 0.25 M aqueous solution of hydrochloric acid with a dose of 5 ml/kg body weight was administered intragastrically with use of a stomach tube. On 28th day, all control and HCL pigs were euthanized and bilateral reght (rNG) and left (lNG) were collected. Cryostat sections were processed for double immunofluorescence using anibodies against SP and NOS. Immunofluorescence staining in the even-numbered ganglia nodes showed the presence of FB-positive cells expressing SP (45,9 ± 3,38% in rNG and 60,4 ± 1,71% in lNG), and nNOS (34,9 ± 6,83% in rNG and 49,9 ± 9,32% in lNG). In HCL group increased expression of both SP (54,8 ± 5,34% in rNG and 56,9 ± 3,28 % in lNG) as well as nNOS (54,9 ± 4,45% in rNG and 52,5 ± 2,17 % in lNG) in FB+ perikaria was found. The acquired results suggest that SP and nNOS are neurotransmitters and/ or neuromodulators participating in the sensory regulation of the prepyloric region of porcine stomach function as well as their potential role in development of the stomach inflamatory state.

Keywords: nNOS, nodose ganglion, pig, SP

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415 Comparison and Effectiveness of Cranial Electrical Stimulation Treatment, Brain Training and Their Combination on Language and Verbal Fluency of Patients with Mild Cognitive Impairment: A Single Subject Design

Authors: Firoozeh Ghazanfari, Kourosh Amraei, Parisa Poorabadi

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Mild cognitive impairment is one of the neurocognitive disorders that go beyond age-related decline in cognitive functions, but in fact, it is not so severe which affects daily activities. This study aimed to investigate and compare the effectiveness of treatment with cranial electrical stimulation, brain training and their double combination on the language and verbal fluency of the elderly with mild cognitive impairment. This is a single-subject method with comparative intervention designs. Four patients with a definitive diagnosis of mild cognitive impairment by a psychiatrist were selected via purposive and convenience sampling method. Addenbrooke's Cognitive Examination Scale (2017) was used to assess language and verbal fluency. Two groups were formed with different order of cranial electrical stimulation treatment, brain training by pencil and paper method and their double combination, and two patients were randomly replaced in each group. The arrangement of the first group included cranial electrical stimulation, brain training, double combination and the second group included double combination, cranial electrical stimulation and brain training, respectively. Treatment plan included: A1, B, A2, C, A3, D, A4, where electrical stimulation treatment was given in ten 30-minutes sessions (5 mA and frequency of 0.5-500 Hz) and brain training in ten 30-minutes sessions. Each baseline lasted four weeks. Patients in first group who first received cranial electrical stimulation treatment showed a higher percentage of improvement in the language and verbal fluency subscale of Addenbrooke's Cognitive Examination in comparison to patients of the second group. Based on the results, it seems that cranial electrical stimulation with its effect on neurotransmitters and brain blood flow, especially in the brain stem, may prepare the brain at the neurochemical and molecular level for a better effectiveness of brain training at the behavioral level, and the selective treatment of electrical stimulation solitude in the first place may be more effective than combining it with paper-pencil brain training.

Keywords: cranial electrical stimulation, treatment, brain training, verbal fluency, cognitive impairment

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414 Application of Thermoplastic Microbioreactor to the Single Cell Study of Budding Yeast to Decipher the Effect of 5-Hydroxymethylfurfural on Growth

Authors: Elif Gencturk, Ekin Yurdakul, Ahmet Y. Celik, Senol Mutlu, Kutlu O. Ulgen

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Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.  

Keywords: COP, HMF, ribosome biogenesis, thermoplastic microbioreactor, yeast

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413 Synthesis and Anticholinesterase Activity of Carvacrol Derivatives

Authors: Fatih Sonmez

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Alzheimer’s disease (AD) is a progressive neurodegenerative disease and it is the most common form of dementia that affects aged people. Acetylcholinesterase is a hydrolase involved in the termination of impulse transmission at cholinergic synapses by rapid hydrolysis of the neurotransmitter ACh in the central and peripheral nervous system. Carvacrol (5-iso-propyl-2-methyl-phenol) is a main bioactive monoterpene isolated from many medicinal herbs, such as Thymus vulgaris, Monarda punctate and Origanum vulgare spp. It is known that carvacrol has been widely used as an active anti-inflammatory ingredient, which can inhibit the isoproterenol induced inflammation in myocardial infarcted rats. In this paper, a series of 12 carvacrol substituted carbamate derivatives (2a-l) was synthesized and their inhibitory activities on AChE and BuChE were evaluated. Among them, 2d exhibited the strongest inhibition against AChE with an IC50 value of 2.22 µM, which was 130-fold more than that of carvacrol (IC50 = 288.26 µM).

Keywords: Acetylcholinesterase, Butyrylcholinesterase, Carbamate, Carvacrol

Procedia PDF Downloads 352
412 Evaluation of Medicinal Plants, Catunaregam spinosa, Houttuynia cordata, and Rhapis excelsa from Malaysia for Antibacterial, Antifungal and Antiviral Properties

Authors: Yik Sin Chan, Bee Ling Chuah, Wei Quan Chan, Ri Jin Cheng, Yan Hang Oon, Kong Soo Khoo, Nam Weng Sit

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Traditionally, medicinal plants have been used to treat different kinds of ailments including infectious diseases. They serve as a good source of lead compounds for the development of new and safer anti-infective agents. This study aimed to investigate the antimicrobial potential of the leaves of three medicinal plants, namely Catunaregam spinosa (Rubiaceae; Mountain pomegranate), Houttuynia cordata (Saururaceae; "fishy-smell herb") and Rhapis excelsa (Arecaceae; “broadleaf lady palm”). The leaves extracts were obtained by sequential extraction using hexane, chloroform, ethyl acetate, ethanol, methanol and water. The antibacterial and antifungal activities were assessed using a colorimetric broth microdilution method against a panel of human pathogenic bacteria (Gram-positive: Bacillus cereus and Staphylococcus aureus; Gram-negative: Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa) and fungi (yeasts: Candida albicans, Candida parapsilosis and Cryptococcus neoformans; Moulds: Aspergillus fumigatus and Trichophyton mentagrophytes) respectively; while antiviral activity was evaluated against the Chikungunya virus on monkey kidney epithelial (Vero) cells by neutral red uptake assay. All the plant extracts showed bacteriostatic activity, however, only 72% of the extracts (13/18) were found to have bactericidal activity. The lowest minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were given by the hexane extract of C. spinosa against S. aureus with the values of 0.16 and 0.31 mg/mL respectively. All the extracts also possessed fungistatic activity. Only the hexane, chloroform and ethyl acetate extracts of H. cordata exerted inhibitory activity against A. fumigatus, giving the lowest fungal susceptibility index of 16.7%. In contrast, only 61% of the extracts (11/18) showed fungicidal activity. The ethanol extract of R. excelsa exhibited the strongest fungicidal activity against C. albicans, C. parapsilosis and T. mentagrophytes with minimum fungicidal concentration (MFC) values of 0.04–0.08 mg/mL, in addition to its methanol extract against T. mentagrophytes (MFC=0.02 mg/mL). For anti-Chikungunya virus activity, only chloroform and ethyl acetate extracts of R. excelsa showed significant antiviral activity with 50% effective concentrations (EC50) of 29.9 and 78.1 g/mL respectively. Extracts of R. excelsa warrant further investigations into their active principles responsible for antifungal and antiviral properties.

Keywords: bactericidal, Chikungunya virus, extraction, fungicidal

Procedia PDF Downloads 403
411 Understanding the Common Antibiotic and Heavy Metal Resistant-Bacterial Load in the Textile Industrial Effluents

Authors: Afroza Parvin, Md. Mahmudul Hasan, Md. Rokunozzaman, Papon Debnath

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The effluents of textile industries have considerable amounts of heavy metals, causing potential microbial metal loads if discharged into the environment without treatment. Aim: In this present study, both lactose and non-lactose fermenting bacterial isolates were isolated from textile industrial effluents of a specific region of Bangladesh, named Savar, to compare and understand the load of heavy metals in these microorganisms determining the effects of heavy metal resistance properties on antibiotic resistance. Methods: Five different textile industrial canals of Savar were selected, and effluent samples were collected in 2016 between June to August. Total bacterial colony (TBC) was counted for day 1 to day 5 for 10-6 dilution of samples to 10-10 dilution. All the isolates were isolated and selected using 4 differential media, and tested for the determination of minimum inhibitory concentration (MIC) of heavy metals and antibiotic susceptibility test with plate assay method and modified Kirby-Bauer disc diffusion method, respectively. To detect the combined effect of heavy metals and antibiotics, a binary exposure experiment was performed, and to understand the plasmid profiling plasmid DNA was extracted by alkaline lysis method of some selective isolates. Results: Most of the cases, the colony forming units (CFU) per plate for 50 ul diluted sample were uncountable at 10-6 dilution, however, countable for 10-10 dilution and it didn’t vary much from canal to canal. A total of 50 Shigella, 50 Salmonella, and 100 E.coli (Escherichia coli) like bacterial isolates were selected for this study where the MIC was less than or equal to 0.6 mM for 100% Shigella and Salmonella like isolates, however, only 3% E. coli like isolates had the same MIC for nickel (Ni). The MIC for chromium (Cr) was less than or equal to 2.0 mM for 16% Shigella, 20% Salmonella, and 17% E. coli like isolates. Around 60% of both Shigella and Salmonella, but only 20% of E.coli like isolates had a MIC of less than or equal to 1.2 mM for lead (Pb). The most prevalent resistant pattern for azithromycin (AZM) for Shigella and Salmonella like isolates was found 38% and 48%, respectively; however, for E.coli like isolates, the highest pattern (36%) was found for sulfamethoxazole-trimethoprim (SXT). In the binary exposure experiment, antibiotic zone of inhibition was mostly increased in the presence of heavy metals for all types of isolates. The highest sized plasmid was found 21 Kb and 14 Kb for lactose and non-lactose fermenting isolates, respectively. Conclusion: Microbial resistance to antibiotics and metal ions, has potential health hazards because these traits are generally associated with transmissible plasmids. Microorganisms resistant to antibiotics and tolerant to metals appear as a result of exposure to metal-contaminated environments.

Keywords: antibiotics, effluents, heavy metals, minimum inhibitory concentration, resistance

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410 Evaluation of Antidiabetic Activity of a Combination Extract of Nigella Sativa & Cinnamomum Cassia in Streptozotocin Induced Type-I Diabetic Rats

Authors: Ginpreet Kaur, Mohammad Yasir Usmani, Mohammed Kamil Khan

Abstract:

Diabetes mellitus is a disease with a high global burden and results in significant morbidity and mortality. In India, the number of people suffering with diabetes is expected to rise from 19 to 57 million in 2025. At present, interest in herbal remedies is growing to reduce the side effects associated with conventional dosage form like oral hypoglycemic agents and insulin for the treatment of diabetes mellitus. Our aim was to investigate the antidiabetic activities of combinatorial extract of N. sativa & C. cassia in Streptozotocin induced type-I Diabetic Rats. Thus, the present study was undertaken to screen postprandial glucose excursion potential through α- glucosidase inhibitory activity (In Vitro) and effect of combinatorial extract of N. sativa & C. cassia in Streptozotocin induced type-I Diabetic Rats (In Vivo). In addition changes in body weight, plasma glucose, lipid profile and kidney profile were also determined. The IC50 values for both extract and Acarbose was calculated by extrapolation method. Combinatorial extract of N. sativa & C. cassia at different dosages (100 and 200 mg/kg orally) and Metformin (50 mg/kg orally) as the standard drug was administered for 28 days and then biochemical estimation, body weights and OGTT (Oral glucose tolerance test) were determined. Histopathological studies were also performed on kidney and pancreatic tissue. In In-Vitro the combinatorial extract shows much more inhibiting effect than the individual extracts. The results reveals that combinatorial extract of N. sativa & C. cassia has shown significant decrease in plasma glucose (p<0.0001), total cholesterol and LDL levels when compared with the STZ group The decreasing level of BUN and creatinine revealed the protection of N. sativa & C. cassia extracts against nephropathy associated with diabetes. Combination of N. sativa & C. cassia significantly improved glucose tolerance to exogenously administered glucose (2 g/kg) after 60, 90 and 120 min interval on OGTT in high dose streptozotocin induced diabetic rats compared with the untreated control group. Histopathological studies shown that treatment with N. sativa & C. cassia extract alone and in combination restored pancreatic tissue integrity and was able to regenerate the STZ damaged pancreatic β cells. Thus, the present study reveals that combination of N. sativa & C. cassia extract has significant α- glucosidase inhibitory activity and thus has great potential as a new source for diabetes treatment.

Keywords: lipid levels, OGTT, diabetes, herbs, glucosidase

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409 Sequential and Combinatorial Pre-Treatment Strategy of Lignocellulose for the Enhanced Enzymatic Hydrolysis of Spent Coffee Waste

Authors: Rajeev Ravindran, Amit K. Jaiswal

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Waste from the food-processing industry is produced in large amount and contains high levels of lignocellulose. Due to continuous accumulation throughout the year in large quantities, it creates a major environmental problem worldwide. The chemical composition of these wastes (up to 75% of its composition is contributed by polysaccharide) makes it inexpensive raw material for the production of value-added products such as biofuel, bio-solvents, nanocrystalline cellulose and enzymes. In order to use lignocellulose as the raw material for the microbial fermentation, the substrate is subjected to enzymatic treatment, which leads to the release of reducing sugars such as glucose and xylose. However, the inherent properties of lignocellulose such as presence of lignin, pectin, acetyl groups and the presence of crystalline cellulose contribute to recalcitrance. This leads to poor sugar yields upon enzymatic hydrolysis of lignocellulose. A pre-treatment method is generally applied before enzymatic treatment of lignocellulose that essentially removes recalcitrant components in biomass through structural breakdown. Present study is carried out to find out the best pre-treatment method for the maximum liberation of reducing sugars from spent coffee waste (SPW). SPW was subjected to a range of physical, chemical and physico-chemical pre-treatment followed by a sequential, combinatorial pre-treatment strategy is also applied on to attain maximum sugar yield by combining two or more pre-treatments. All the pre-treated samples were analysed for total reducing sugar followed by identification and quantification of individual sugar by HPLC coupled with RI detector. Besides, generation of any inhibitory compounds such furfural, hydroxymethyl furfural (HMF) which can hinder microbial growth and enzyme activity is also monitored. Results showed that ultrasound treatment (31.06 mg/L) proved to be the best pre-treatment method based on total reducing content followed by dilute acid hydrolysis (10.03 mg/L) while galactose was found to be the major monosaccharide present in the pre-treated SPW. Finally, the results obtained from the study were used to design a sequential lignocellulose pre-treatment protocol to decrease the formation of enzyme inhibitors and increase sugar yield on enzymatic hydrolysis by employing cellulase-hemicellulase consortium. Sequential, combinatorial treatment was found better in terms of total reducing yield and low content of the inhibitory compounds formation, which could be due to the fact that this mode of pre-treatment combines several mild treatment methods rather than formulating a single one. It eliminates the need for a detoxification step and potential application in the valorisation of lignocellulosic food waste.

Keywords: lignocellulose, enzymatic hydrolysis, pre-treatment, ultrasound

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408 Evaluation of Antimicrobial and Anti-Inflammatory Activity of Doani Sidr Honey and Madecassoside against Propionibacterium Acnes

Authors: Hana Al-Baghaoi, Kumar Shiva Gubbiyappa, Mayuren Candasamy, Kiruthiga Perumal Vijayaraman

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Acne is a chronic inflammatory disease of the sebaceous glands characterized by areas of skin with seborrhea, comedones, papules, pustules, nodules, and possibly scarring. Propionibacterium acnes (P. acnes), plays a key role in the pathogenesis of acne. Their colonization and proliferation trigger the host’s inflammatory response leading to the production of pro-inflammatory cytokines such as interleukin-8 (IL-8) and tumour necrosis factor-α (TNF-α). The usage of honey and natural compounds to treat skin ailments has strong support in the current trend of drug discovery. The present study was carried out evaluate antimicrobial and anti-inflammatory potential of Doani Sidr honey and its fractions against P. acnes and to screen madecassoside alone and in combination with fractions of honey. The broth dilution method was used to assess the antibacterial activity. Also, ultra structural changes in cell morphology were studied before and after exposure to Sidr honey using transmission electron microscopy (TEM). The three non-toxic concentrations of the samples were investigated for suppression of cytokines IL 8 and TNF α by testing the cell supernatants in the co-culture of the human peripheral blood mononuclear cells (hPBMCs) heat killed P. acnes using enzyme immunoassay kits (ELISA). Results obtained was evaluated by statistical analysis using Graph Pad Prism 5 software. The Doani Sidr honey and polysaccharide fractions were able to inhibit the growth of P. acnes with a noteworthy minimum inhibitory concentration (MIC) value of 18% (w/v) and 29% (w/v), respectively. The proximity of MIC and MBC values indicates that Doani Sidr honey had bactericidal effect against P. acnes which is confirmed by TEM analysis. TEM images of P. acnes after treatment with Doani Sidr honey showed completely physical membrane damage and lysis of cells; whereas non honey treated cells (control) did not show any damage. In addition, Doani Sidr honey and its fractions significantly inhibited (> 90%) of secretion of pro-inflammatory cytokines like TNF α and IL 8 by hPBMCs pretreated with heat-killed P. acnes. However, no significant inhibition was detected for madecassoside at its highest concentration tested. Our results suggested that Doani Sidr honey possesses both antimicrobial and anti-inflammatory effects against P. acnes and can possibly be used as therapeutic agents for acne. Furthermore, polysaccharide fraction derived from Doani Sidr honey showed potent inhibitory effect toward P. acnes. Hence, we hypothesize that fraction prepared from Sidr honey might be contributing to the antimicrobial and anti-inflammatory activity. Therefore, this polysaccharide fraction of Doani Sidr honey needs to be further explored and characterized for various phytochemicals which are contributing to antimicrobial and anti-inflammatory properties.

Keywords: Doani sidr honey, Propionibacterium acnes, IL-8, TNF alpha

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407 Synthesis of Some 1h-Benzimidazoles as Inhibitors of EGFR Tyrosine Kinase

Authors: İsmail Çeli̇k, Gülgün Ayhan-Kılcıgi̇l, Arzu Onay-Beşi̇kçi̇

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In this study, some 2-(2-phenyl/substitutedphenyl)- lH-benzo[d]'imidazol-l-yl)-N'-(alkylthiosemicarbazide were designed and prepared. Firstly, 2-phenyl/ suhstitutedphenyl-lH-Benzo[d]imidazole was prepared via oxidative condensation of o-phenylenediamine, benzaldehyde and sodium metabisulfite. Treatment of the benzimidazole compound with ethyl chloroacetate in KOH/DMSO gave the ester compound ethyl 2-(2-substitutedphenyl)-1H-benzo[d]imidazol-l-yl)acetate. Hydrazine hydrate and the ester in ethanol were refluxed for 4 h to give 2-(2-phenyl/substitutedphenyl)-1H-benzo[d]imidazol-l-yl)acetohydrazide. Thiosemicarbazides were obtained by condensing acyl hydrazide with the alkylisothiocyanate in ethanol. Following the structure elucidation, benzimidazole compounds were tested for their EGFR kinase inhibitory activities by using ADP-GloTM Kinase Assay.

Keywords: benzimidazole, EGFR kinase inhibitor, synthesis, thiosemicarbazide

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406 The Effect of Music on Consumer Behavior

Authors: Lara Ann Türeli, Özlem Bozkurt

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There is a biochemical component to listening to music. The type of music listened to can lead to different levels of neurotransmitter and biochemical activity within the brain, resulting in brain stimulation and different moods. Therefore, music plays an important role in neuromarketing and consumer behavior. The quality of a commercial can be measured by the effect the music has on its audience. Thus, understanding how music can affect the brain can provide better marketing strategies for all businesses. The type of music used plays an important role in how a person responds to certain experiences. In the context of marketing and consumer behavior, music can determine whether a person will be intrigued to buy something. Depending on the type of music listened to by an individual; the music may trigger the release of pleasurable neurotransmitters such as dopamine. Dopamine is a neurotransmitter that plays an important role in reward pathways in the brain. When an individual experiences a pleasurable activity, increased levels of dopamine are produced, eventually leading to the formation of new reward pathways. Consequently, the increased dopamine activity within the brain triggered by music can result in new reward pathways along the dopamine pathways in the brain. Selecting pleasurable music for commercials can result in long-term brain stimulation, increasing consumerism. The effect of music on consumerism should be considered not only in commercials but also in the atmosphere it creates within stores. The type of music played in a store can affect consumer behavior and intention. Specifically, the rhythm, pitch, and pace of music can contribute to the mood of the song. The background music in a store can determine the consumer’s emotional presence and consequently affect their intentions. In conclusion, understanding the physiological, psychological, and neurochemical basis of the effect of music on brain stimulation is essential to understand consumer behavior. The role of dopamine in the formation of reward pathways as a result of music directly contributes to consumer behavior and the tendency of a commercial or store to leave a long-term effect on the consumer. The careful consideration of the pitch, pace, and rhythm of a song in the selection of music can not only help companies predict the behavior of a consumer but also determine the behavior of a consumer.

Keywords: sensory processing, neuropsychology, dopamine, neuromarketing

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405 Effect of Melatonin on Seed Germination and Seedling Growth of Catharanthus roseus under Cadmium Stress

Authors: Rayhaneh Amooaghaie, Masoomeh Nabaei

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In this study, 200 µM Cd reduced relative seed germination, root elongation tolerance and seed germination tolerance index of Catharanthus roseus. The melatonin improved seed germination, germination velocity, seedling length and vigor index under Cd stress in a dose-dependent manner and the maximum biological responses obtained by 100 μM melatonin. However, 200-400 μM melatonin and 400 μM SNP had negative effects that evidenced as lower germination indices and poor establishment of seedlings. The cadmium suppressed amylase activity and contents of soluble and reducing sugars in germinating seeds, thereby reduced seed germination and subsequent seedling growth whereas increased electrolyte leakage. These Cd-induced inhibitory effects were ameliorated by melatonin.

Keywords: cadmium, Catharanthus roseus, melatonin, seed germination

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404 Bismuth-Inhibitory Effects on Bacteria and Stimulation of Fungal Growth In vitro

Authors: Sulaiman B. Ali Alharbi, Bassam H. Mashat, Naif Abdullah Al-Harbi, Milton Wainwright, Abeer S. Aloufi, Sulamain Alnaimat

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Bismuth salicylate was found to inhibit the growth of a range of bacteria and yeast, Candida albican. In general the growth of bacteria did not result in the increase in bismuth solubilisation, in contrast, bismuth solubilisation increased following the growth of C. albicans. A significant increase in the biomass (dry weight) of Aspergillus niger and Aspergillus oryzae occurred in vitro when these fungi were grown in the presence of bismuth salicylate. Biomass increase occurred over a range of bismuth compound additions, which in the case of A. oryzae was associated with the increase in the solubilisation of the insoluble bismuth compounds.

Keywords: bacterial inhibition, fungal growth stimulation, medical uses of bismuth, yeast inhibition

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403 Effect of Antimony on Microorganisms in Aerobic and Anaerobic Environments

Authors: Barrera C. Monserrat, Sierra-Alvarez Reyes, Pat-Espadas Aurora, Moreno Andrade Ivan

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Antimony is a toxic and carcinogenic metalloid considered a pollutant of priority interest by the United States Environmental Protection Agency. It is present in the environment in two oxidation states: antimonite (Sb (III)) and antimony (Sb (V)). Sb (III) is toxic to several aquatic organisms, but the potential inhibitory effect of Sb species for microorganisms has not been extensively evaluated. The fate and possible toxic impact of antimony on aerobic and anaerobic wastewater treatment systems are unknown. For this reason, the objective of this study was to evaluate the microbial toxicity of Sb (V) and Sb (III) in aerobic and anaerobic environments. Sb(V) and Sb(III) were used as potassium hexahydroxoantimonate (V) and potassium antimony tartrate, respectively (Sigma-Aldrich). The toxic effect of both Sb species in anaerobic environments was evaluated on methanogenic activity and the inhibition of hydrogen production of microorganisms from a wastewater treatment bioreactor. For the methanogenic activity, batch experiments were carried out in 160 mL serological bottles; each bottle contained basal mineral medium (100 mL), inoculum (1.5 g of VSS/L), acetate (2.56 g/L) as substrate, and variable concentrations of Sb (V) or Sb (III). Duplicate bioassays were incubated at 30 ± 2°C on an orbital shaker (105 rpm) in the dark. Methane production was monitored by gas chromatography. The hydrogen production inhibition tests were carried out in glass bottles with a working volume of 0.36 L. Glucose (50 g/L) was used as a substrate, pretreated inoculum (5 g VSS/L), mineral medium and varying concentrations of the two species of antimony. The bottles were kept under stirring and at a temperature of 35°C in an AMPTSII device that recorded hydrogen production. The toxicity of Sb on aerobic microorganisms (from a wastewater activated sludge treatment plant) was tested with a Microtox standardized toxicity test and respirometry. Results showed that Sb (III) is more toxic than Sb (V) for methanogenic microorganisms. Sb (V) caused a 50% decrease in methanogenic activity at 250 mg/L. In contrast, exposure to Sb (III) resulted in a 50% inhibition at a concentration of only 11 mg/L, and an almost complete inhibition (95%) at 25 mg/L. For hydrogen-producing microorganisms, Sb (III) and Sb (V) inhibited 50% of this production with 12.6 mg/L and 87.7 mg/L, respectively. The results for aerobic environments showed that 500 mg/L of Sb (V) do not inhibit the Allivibrio fischeri (Microtox) activity or specific oxygen uptake rate of activated sludge. In the case of Sb (III), this caused a loss of 50% of the respiration of the microorganisms at concentrations below 40 mg/L. The results obtained indicate that the toxicity of the antimony will depend on the speciation of this metalloid and that Sb (III) has a significantly higher inhibitory potential compared to Sb (V). It was shown that anaerobic microorganisms can reduce Sb (V) to Sb (III). Acknowledgments: This work was funded in part by grants from the UA-CONACYT Binational Consortium for the Regional Scientific Development and Innovation (CAZMEX), the National Institute of Health (NIH ES- 04940), and PAPIIT-DGAPA-UNAM (IN105220).

Keywords: aerobic inhibition, antimony reduction, hydrogen inhibition, methanogenic toxicity

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402 Assessing the Antimicrobial Activity of Chitosan Nanoparticles by Fluorescence-Labeling

Authors: Laidson P. Gomes, Cristina T. Andrade, Eduardo M. Del Aguila, Cameron Alexander, Vânia M. F. Paschoalin

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Chitosan is a natural polysaccharide prepared by the N-deacetylation of chitin. In this study, the physicochemical and antibacterial properties of chitosan nanoparticles, produced by ultrasound irradiation, were evaluated. The physicochemical properties of the nanoparticles were determined by dynamic light scattering and zeta potential analysis. Chitosan nanoparticles inhibited the growth of E. coli. The minimum inhibitory concentration (MIC) values were lower than 0.5 mg/mL, and the minimum bactericidal concentration (MBC) values were similar or higher than MIC values. Confocal laser scanning micrographs (CLSM) were used to observe the interaction between E. coli suspensions mixed with FITC-labeled chitosan polymers and nanoparticles.

Keywords: chitosan nanoparticles, dynamic light scattering, zeta potential, confocal microscopy, antibacterial activity

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401 Antioxydant Activity of Flavonoïd’s Extracts of Rhamnus alaternus L. Leaves of Tessala Mountains (Occidental Algeria)

Authors: Benchiha Walid, Mahroug Samira

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Rhamnus alaternus L. is a shrub that belongs to the family of Rhamnaceae. It is a medicinal plant that is largely used in traditional medicine in Algeria. Five flavonoic extracts obtained of Rhamnus alaternus L. leaves. The flavonoids were evaluated by a method that uses aluminum chloride AlCl3 of each extract; the content is estimated at 19.33 (Hexanic. Extract), 18.42 (Chlroformic.extract), 16.75 (Acetate. Extract), 3.9 (Brute. Extract), and 3.02 (Aqueous. Extract) mg Equivalent quercetine/gram of extract (mg QE/ g extract). The antioxidant activity was realized by the antiradical test that was evaluated by using DPHH (2.2 diphenyl-1-1picrylhdrazile), the inhibitory concentration at 50% (CI50) were estimated at 74.78 (Vitamin.C), 143.78 (Catechine), 101.78 (Gallic acid), 205.41 (Tannic acid), 210 (Caffeic acid) µg/ml; 74.16 (Br.extr), 9.98 (Aq.extr), 54.08 (Hèx.extr), 8.64 (Ac.extr), 30.49 (Ch.extr) mg/ml.

Keywords: Rhamnus alaternus L., flavonoids, antioxydant activity, Tessala

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400 Quantitative Structure Activity Relationship Model for Predicting the Aromatase Inhibition Activity of 1,2,3-Triazole Derivatives

Authors: M. Ouassaf, S. Belaidi

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Aromatase is an estrogen biosynthetic enzyme belonging to the cytochrome P450 family, which catalyzes the limiting step in the conversion of androgens to estrogens. As it is relevant for the promotion of tumor cell growth. A set of thirty 1,2,3-triazole derivatives was used in the quantitative structure activity relationship (QSAR) study using regression multiple linear (MLR), We divided the data into two training and testing groups. The results showed a good predictive ability of the MLR model, the models were statistically robust internally (R² = 0.982) and the predictability of the model was tested by several parameters. including external criteria (R²pred = 0.851, CCC = 0.946). The knowledge gained in this study should provide relevant information that contributes to the origins of aromatase inhibitory activity and, therefore, facilitates our ongoing quest for aromatase inhibitors with robust properties.

Keywords: aromatase inhibitors, QSAR, MLR, 1, 2, 3-triazole

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399 Extract and Naphthoquinone Derivatives from in vitro Culture of an Ascomycetous Marine Fungus with Antibacterial Activity

Authors: Uftah Ali M. Shushni, Viola Stuppec, Ulrike Lindequist

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Because of the evolving resistance of microorganisms to existing antibiotics, there is an increasing need for new antibiotics not only in human but also in veterinary medicine. As part of our ongoing work on the secondary metabolites produced by marine fungi, the organic extract of the culture filtrate of an Ascomycetous fungus, which was found on driftwood collected from the coast of the Greifswalder Bodden, Baltic Sea, Germany displayed antimicrobial activity against some fish and human pathogenic bacteria. Bioactivity-guided column chromatographic separation led to the isolation of 6-Deoxybostrycoidin. The structure was determined from the interpretation of spectroscopic data (UV, MS, and NMR). 6-Deoxybostrycoidin exhibited in vitro activity against Bacillus subtilis, Staphylococcus aureus and Flexibacter maritimus with minimal inhibitory concentrations of 25, 12.5 and 12.5 μg/ml respectively.

Keywords: marine fungi, fish pathogenic bacteria, microorganism, medicine

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398 Extractive Fermentation of Ethanol Using Vacuum Fractionation Technique

Authors: Weeraya Samnuknit, Apichat Boontawan

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A vacuum fractionation technique was introduced to remove ethanol from fermentation broth. The effect of initial glucose and ethanol concentrations were investigated for specific productivity. The inhibitory ethanol concentration was observed at 100 g/L. In order to increase the fermentation performance, the ethanol product was removed as soon as it is produced. The broth was boiled at 35°C by reducing the pressure to 65 mBar. The ethanol/water vapor was fractionated for up to 90 wt% before leaving the column. Ethanol concentration in the broth was kept lower than 25 g/L, thus minimized the product inhibition effect to the yeast cells. For batch extractive fermentation, a high substrate utilization rate was obtained at 26.6 g/L.h and most of glucose was consumed within 21 h. For repeated-batch extractive fermentation, addition of glucose was carried out up to 9 times and ethanol was produced more than 8-fold higher than batch fermentation.

Keywords: ethanol, extractive fermentation, product inhibition, vacuum fractionation

Procedia PDF Downloads 250