Search results for: D-amino Acid Oxidase (DAO) enzyme

Authors: Emmanuel C. Ngerem

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The ever-increasing depletion of the dominant global form of energy (fossil fuels) calls for the development of sustainable and green alternative energy sources such as bioethanol, biohydrogen, and biodiesel. The production of the major biofuels relies on biomass feedstocks that are mainly derived from edible food crops and some inedible plants. One suitable feedstock with great potential as raw material for biofuel production is microalgal biomass. Despite the tremendous attributes of microalgae as a source of biofuel, their cultivation requires huge volumes of freshwater, thus posing a serious threat to commercial-scale production and utilization of algal biomass. In this study, a multi-media wastewater mixture for microalgae growth was formulated and optimized. Moreover, the obtained microalgae biomass was pre-treated to reduce sugar recovery and was compared with previous studies on microalgae biomass pre-treatment. The formulated and optimized mixed wastewater media for biomass and lipid accumulation was established using the simplex lattice mixture design. Based on the superposition approach of the potential results, numerical optimization was conducted, followed by the analysis of biomass concentration and lipid accumulation. The coefficients of regression (R²) of 0.91 and 0.98 were obtained for biomass concentration and lipid accumulation models, respectively. The developed optimization model predicted optimal biomass concentration and lipid accumulation of 1.17 g/L and 0.39 g/g, respectively. It suggested 64.69% dairy wastewater (DWW) and 35.31% paper and pulp wastewater (PWW) mixture for biomass concentration, 34.21% DWW, and 65.79% PWW for lipid accumulation. Experimental validation generated 0.94 g/L and 0.39 g/g of biomass concentration and lipid accumulation, respectively. The obtained microalgae biomass was pre-treated, enzymatically hydrolysed, and subsequently assessed for reducing sugars. The optimization of microwave pre-treatment of Chlorococcum sp. was achieved using response surface methodology (RSM). Microwave power (100 – 700 W), pre-treatment time (1 – 7 min), and acid-liquid ratio (1 – 5%) were selected as independent variables for RSM optimization. The optimum conditions were achieved at microwave power, pre-treatment time, and acid-liquid ratio of 700 W, 7 min, and 32.33:1, respectively. These conditions provided the highest amount of reducing sugars at 10.73 g/L. Process optimization predicted reducing sugar yields of 11.14 g/L on microwave-assisted pre-treatment of 2.52% HCl for 4.06 min at 700 watts. Experimental validation yielded reducing sugars of 15.67 g/L. These findings demonstrate that dairy wastewater and paper and pulp wastewater that could pose a serious environmental nuisance. They could be blended to form a suitable microalgae growth media, consolidating the potency of microalgae as a viable feedstock for fermentable sugars. Also, the outcome of this study supports the microalgal wastewater biorefinery concept, where wastewater remediation is coupled with bioenergy production.

Keywords: wastewater cultivation, mixture design, lipid, biomass, nutrient removal, microwave, Chlorococcum, raceway pond, fermentable sugar, modelling, optimization

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Authors: N. Hasbullah, K. A. Sekak

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The sulfonated poly (ether ether ketone) SPEEK nanofiber membrane were successfully electrospun for Polymer Electrolyte Membrane (PEM) in Proton Exchange Membrane Fuel Cell (PEMFC) and their nanosized properties were investigated. The poly (ether ether ketone) PEEK victrex® grade 90p was sulfonated with concentrated sulfuric acid (95-98% w/w) at room temperature for 60 hours sulfonation times. The degree sulfonation of SPEEK are 70% was determined by H1 NMR and the functional groups of the SPEEK were characterize using FTIR. Then, the SPEEK nanofiber membrane were prepared via electrospinning method using DMAC as a solvent. The SPEEK sample were successfully electrospun using predetermine set up. FESEM show the electrospun fiber mat surface and confirmed the nanostructure membrane cell.

Keywords: polymer electrolyte membrane (PEM), sulfonated poly (ether ether ketone) (SPEEK), degree sulfonation, Electrospinning, Nanofibers

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Authors: Akintayo Emmanuel Temitope, Akintayo Cecilia Olufunke, Ziegler Thomas

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Bio-polyols are important components in polyurethane industries. The preliminary studies into the synthesis of bio-polyol products (epoxy-polyol and epoxyl-polyurethanes) from Jatropha curcas were investigated. The reactions were followed by both infrared and nuclear magnetic resonance. Physico-chemical characterisation of the samples for iodine value (IV), acid value (AV), saponification value (SV) and hydroxyl value (HV) were carried out. Thermal transitions of the products were studied by heating 5 mg of the sample from 20ºC to 800ºC and then cooling down to -500ºC on a differential scanning calorimeter (DSC). The preparation of epoxylpolyol and polyurethane from Jatropha curcas oil was smooth and efficient. Results of film and solubility properties revealed that coatings of Jatropha curcas epoxy-polyurethanes performed better with increased loading of toluylene 2, 4-diisocyanate (TDI) up to 2 wt% while their solvent resistance decreased beyond a TDI loading of 1.2 wt%. DSC analysis shows the epoxy-polyurethane to be less stable compared to the epoxy-polyol.

Keywords: synthesis, epoxy-polyol, epoxy-polyurethane, jatropha curcas oil

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Authors: Bhojan Dhakal, Naba Raj Devkota, Chet Raj Upreti, Maheshwar Sapkota

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A study was conducted to find out the production potential, nutrient composition, and the variability of the most commonly available fodder trees along with the varying altitude to help optimize the dry matter requirement during winter lean period. The study was carried out from March to June, 2012 in Lho and Prok Village Development Committee of Manaslu Conservation Area (MCA), located in Gorkha district of Nepal. The other objective of the research was to learn the impact of climate change on livestock production linking it with feed availability. The study was conducted in two parts: social and biological. Accordingly, a households (HHs) survey was conducted to collect primary data from 70 HHs, focusing on the perception of respondents on impacts of climatic variability on the feeding management. The next part consisted of understanding yield potential and nutrient composition of the four most commonly available fodder trees (M. azedirach, M. alba, F. roxburghii, F. nemoralis), within two altitudes range: (1500-2000 masl and 2000-2500 masl) by using a RCB design in 2*4 factorial combination of treatments, each replicated four times. Results revealed that majority of the farmers perceived the change in climatic phenomenon more severely within the past five years. Farmers were using different adaptation technologies such as collection of forage from jungle, reducing unproductive animals, fodder trees utilization, and crop by product feeding at feed scarcity period. Ranking of the different fodder trees on the basis of indigenous knowledge and experiences revealed that F. roxburghii was the best-preferred fodder tree species (index value 0.72) in terms overall preferability whereas M. azedirach had highest growth and productivity (index value 0.77), F. roxburghii had highest adoptability (index value 0.69) and palatability (index value 0.69) as well. Similarly, fresh yield and dry matter yield of the each fodder trees was significant (P < 0.01) between the altitude and within species. Fodder trees yield analysis revealed that the highest dry matter (DM) yield (28 kg/tree) was obtained for F. roxburghii but that remained statistically similar (P > 0.05) to the other treatment. On the other hand, most of the parameters: ether extract (EE), acid detergent lignin (ADL), acid detergent fibre (ADF), cell wall digestibility (CWD), relative digestibility (RD), digestible nutrient (TDN), and Calcium (Ca) among the treatments were highly significant (P < 0.01). This indicates the scope of introducing productive and nutritive fodder trees species even at the high altitude to help reduce fodder scarcity problem during winter. The finding also revealed the scope of promoting all available local fodder trees species as crude protein content of these species were similar.

Keywords: fodder trees, yield potential, climate change, nutrient composition

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Authors: Naoual Boussoualim, Hayat Trabsa, Iman Krache, Lekhmici Arrar, Abderrahmane Baghiani

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Resistance to antibiotics has emerged following their widespread use; the important mechanism of beta-lactam resistance in bacteria is the production of beta-lactamase. In order to find new bioactive beta-lactamase inhibitors, this study investigated the inhibition effect of the extracts of Anchusa azurea (AA) on a beta-lactamase from Bacillus cereus. The extracts exerted inhibitory effects on beta-lactamase in a dose-dependent manner, the results showed that the crude extract (BrE) and the ethyl acetate extract (AcE) of Anchusa azurea showed a very high inhibitory activity at a concentration of 10 mg, the percentage of inhibition was between 58% and 68%. Not all extracts were as potent as the original inhibitors such as clavulanic acid, the isolation and the structural elucidation of the active constituents in these extracts will provide useful means in the development of beta -lactamase inhibitors.

Keywords: Anchusa azurea, natural product, resistance, antibiotics, beta-lactamase, inhibitors

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Authors: Gafar Bamigbade, Adebunkola Omemu

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“Ogi” is a fermented cereal gruel prepared from maize (Zea mays), millet (Pennisetum typhoideum) or guinea corn (Sorghum bicolour). It could be boiled to give a thicker consistency wrapped in leaf allowed to cool and set to a gel known as “eko”. The objective of this study is to determine the amylase activities of mould associated with the spoilage of Ogi and eko. Moulds were isolated from spoilt Ogi and eko samples using standard microbiological procedures. The isolate was then screened for amylase production using starch agar medium. Positive isolates were used for amylase production by solid state fermentation (SFF) using rice bran as the medium. An alpha-amylase and glucoamylase activity of the crude enzyme was determined using the DNS method. The mean mold Population ranged from 1.15 X 105cfu/g for raw Ogi to 6.25 X 105cfu/g for Eko (wrapped in Leaves). Twenty-seven (27) moulds isolated from the sample include A. niger, A. flavus, A. fumigatus, Rhizopus species and Penicillium species. Aspergillus flavus had the highest percentage (51.9%) of incidence while Penicillium species had the least (3.7%). Out of the 27 isolates screened, 19 were found to be amylase positive by showing a clear zone around their colony after flooding with iodine solution. Diameter of clear zone ranged from 3.00mm (Aspergillus niger, C4) to 22.00mm (Aspergillus flavus, A1). Aspergillus niger isolated from spoilt Eko wrapped in leaf has the highest percentage alpha-amylase activity (30.8%) and Aspergillus flavus isolated from spoilt raw ogi has the lowest activity (11.4%). Aspergillus niger isolated from spoilt Eko wrapped in nylon produces the highest glucoamylase activity (240U/ml) while penicillium specie isolated from spoilt cooked ogi has the lowest activity (100U/ml). This study shows that moulds associated with spoilage of ogi and eko can produce amylase.

Keywords: glucoamylase, alpha amylase, ogi, eko

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Authors: Pardis Abolghasemi, Benyamin Hatamsaz

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Background: Diabetic nephropathy is a serious health problem described by specific kidney structure and functional disturbance. Renoprotective effects of the stem cells secretase have been shown in many kidney diseases. The aim is to evaluate the capability of human Wharton’s jelly mesenchymal stem cells conditioned media (hWJMSCs-CM) to alleviate DN in streptozotocin (STZ)-induced diabetes. Methods: Diabetic nephropathy was induced by injection of STZ (60 mg/kg, IP) in twenty rats. Conditioned media was extracted from hWJMSCs at third passages. At week 8, diabetic rats were divided into two groups: treated (hWJMSCs-CM, 500 μl/rat for three weeks, IP) and not treated (DN). In the 11th week, three groups (control, DN and DN+hWJMSCs-CM) were kept in metabolic cages and urine was collected for 24h. Blood pressure (BP) and heart rate (HR) were continuously recorded. The serum samples were maintained for measuring BUN, Cr and angiotensin-converting enzyme (ACE) activity. The left kidney was kept at -80°C for ACE activity assessment. The right kidney and pancreas were used for histopathologic evaluation. Result: Diabetic nephropathy was detected by microalbuminuria and increased albumin/creatinine ratio, as well as the pancreas and renal structural disturbance. Glomerular filtration rate, BP and HR increased in the DN group. The ACE activity was elevated in the serum and kidneys of the DN group. Administration of hWJMSCs-CM modulated the renal functional and structural disturbance and decreased the ACE activity. Conclusion: Conditioned media was extracted from hWJMSCs may have a Renoprotective effect in diabetic nephropathy. This may happen through regulation of ACE activity and renin-angiotensin system inhibition.

Keywords: diabetic nephropathy, mesenchymal stem cells, immunomodulation, anti-inflammation

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Authors: Radka Obořilová, Hana Šimečková, Matěj Pastucha, Jan Přibyl, Petr Skládal, Ivana Mašlaňová, Zdeněk Farka

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Due to the spreading of antimicrobial resistance, alternative approaches to combat superinfections are being sought, both in the field of lysing agents and methods for studying bacterial lysis. A suitable alternative to antibiotics is phage therapy and enzybiotics, for which it is also necessary to study the mechanism of their action. Biosensor-based techniques allow rapid detection of pathogens in real time, verification of sensitivity to commonly used antimicrobial agents, and selection of suitable lysis agents. The detection of lysis takes place on the surface of the biosensor with immobilized bacteria, which has the potential to be used to study biofilms. An example of such a biosensor is surface plasmon resonance (SPR), which records the kinetics of bacterial lysis based on a change in the resonance angle. The bacteria are immobilized on the surface of the SPR chip, and the action of phage as the mass loss is monitored after a typical lytic cycle delay. Atomic force microscopy (AFM) is a technique for imaging of samples on the surface. In contrast to electron microscopy, it has the advantage of real-time imaging in the native conditions of the nutrient medium. In our case, Staphylococcus aureus was lysed using the enzyme lysostaphin and phage P68 from the familyPodoviridae at 37 ° C. In addition to visualization, AFM was used to study changes in mechanical properties during lysis, which resulted in a reduction of Young’s modulus (E) after disruption of the bacterial wall. Changes in E reflect the stiffness of the bacterium. These advanced methods provide deeper insight into bacterial lysis and can help to fight against bacterial diseases.

Keywords: biosensors, atomic force microscopy, surface plasmon resonance, bacterial lysis, staphylococcus aureus, phage P68

Procedia PDF Downloads 131

Authors: Eduzor Esther, Uhiara Ngozi, Ya’u Abubakar Umar, Anayo Jacob Gabriel, Umar Ahmed

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The study investigated the characteristic effect of treating okra seed and soybeans seed oil with crude enzymes extract from malted rice. The oils from okra seeds and soybeans were obtained by solvent extraction method using N-hexane solvent. Soybeans seeds had higher percentage oil yield than okra seed. 250ml of each oil was thoroughly mixed with 5ml of the malted rice extract at 400C for 5mins and then filtered and regarded as treated oil while another batch of 250ml of each oil was not mixed with the malted rice extract and regarded as untreated oil. All the oils were analyzed for specific gravity, refractive index, emulsification capacity, absortivity, TSS and viscosity. Treated okra seed and soybeans oil gave higher values for specific gravity, than the untreated oil for okra seed and soybeans oil respectively. The emulsification capacity values were also higher for treated oils, when compared to the untreated oil, for okra seed and soybeans oil respectively. Treated okra seed and soybeans oil also had higher range of values for absorptivity, than the untreated oil for okra seed and soybeans respectively. The ranges of T.S.S values of the treated oil were also higher, than those of the untreated oil for okra seed and soybeans respectively. The results of viscosity showed that the treated oil had higher values, than the untreated oil for okra seed and soybeans oil respectively. However, the results of refractive index showed that the untreated oils had higher values ranges of than the treated oils for okra seed and soybeans respectively. Treated oil show better quality in respect to the parameters analyst, except the refractive index which is slightly less but also is within the rangiest of standard, the oils are high in unsaturation especially okra oil when compared with soya beans oil. It is recommended that, treated oil of okra seeds and soya beans can serve better than many oils that presently in use such as ground nut oil, palm oil and cotton seeds oil.

Keywords: extract, malted, oil, okra, rice, seed, soybeans

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Authors: Özge Selin Çevik, Leyla Şahin, Gülhan Örekeci Temel

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The early postnatal period is critical for the development of cognitive and emotional functions. Maternal separation is a detrimental postnatal influence, whereas environmental enrichment is a therapeutic and protective agent. It is unclear if long-term environmental enrichment can compensate for the effects of maternal separation stress on anxiety behavior. This study was designed to examine how environmental enrichment affects anxiety levels and corticosterone levels in maternally separated rats. There are six main groups in this study: control (C), maternal separation+standard cage (MS), maternal separation+enriched environment (MSE), enriched environment (E), the maternal separation that decapitated at postnatal (PN) 21 (MS21), and standard cage that decapitated at PN21 (STD21). The maternal separation procedure consisted of PN for 21 days (between 09:00 a.m and 12:00 a.m). Enriched (E, MSE) or standard cage environment rats (MS, C) spent PN (22-55) days in either enriched cages or standard cages. Anxiety and locomotor activity were examined with the open field and elevated plus-maze test. Blood corticosterone level was evaluated by the enzyme-linked immunosorbent assay (ELISA) method. Results showed that maternal separation (MS) increased locomotor activity and anxiety. An enriched environment (E) did not change the locomotor activity. MSE group’s anxiety and locomotor activity did not change. Corticosterone levels increased in the maternal separation group that decapitated at the PN 21 days. Maternal separation increases anxiety. Environmental enrichment alone was insufficient to cause alterations in the anxiety level. In addition, environmental enrichment did not ameliorate the anxiety level in maternally separated rats. However, environmental enrichment decreased the locomotor activity in the maternally separated rats.

Keywords: maternal separation, environment enrichment, stress, hippocampus, anxiety, memory, rat

Procedia PDF Downloads 87

Authors: Kritsada Pipitthapan

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WO3/SiO2 catalysts were modified by an ion exchange method with sodium hydroxide or potassium hydroxide solution. The performance of the modified catalysts was tested in the metathesis of ethylene and trans-2-butene to propylene. During ion exchange, sodium and potassium ions played different roles. Sodium modified catalysts revealed constant trans-2-butene conversion and propylene selectivity when the concentrations of sodium in the solution were varied. In contrast, potassium modified catalysts showed reduction of the conversion and increase of the selectivity. From these results, potassium hydroxide may affect the transformation of tungsten oxide active species, resulting in the decrease in conversion whereas sodium hydroxide did not. Moreover, the modification of catalysts by this method improved the catalyst stability by lowering the amount of coke deposited on the catalyst surface.

Keywords: acid sites, alkali metal, isomerization, metathesis

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Authors: M. S. Matlala, I. Ignatious

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Environmental sustainability has taken the center stage in human life all over the world. Energy is the most essential component of our life. The conventional sources of energy are non-renewable and have a detrimental environmental impact. Therefore, there is a need to move from conventional to non-conventional renewable energy sources to satisfy the world’s energy demands. The study aimed at screening for microbial cellulolytic enzymes using a proteomic approach. The objectives were to screen for microbial cellulases with high specific activity and separate the cellulolytic enzymes using a combination of zymography and two-dimensional (2-D) gel electrophoresis followed by tryptic digestion, Matrix-assisted Laser Desorption Ionisation-Time of Flight (MALDI-TOF) and bioinformatics analysis. Fungal and bacterial isolates were cultured in M9 minimal and Mandel media for a period of 168 hours at 60°C and 30°C with cellobiose and Avicel as carbon sources. Microbial cells were separated from supernatants through centrifugation, and the crude enzyme from the cultures was used for the determination of cellulase activity, zymography, SDS-PAGE, and two-dimensional gel electrophoresis. Five isolates, with lytic action on carbon sources studied, were a bacterial strain (BARK) and fungal strains (VCFF1, VCFF14, VCFF17, and VCFF18). Peak cellulase production by the selected isolates was found to be 3.8U/ml, 2.09U/ml, 3.38U/ml, 3.18U/ml, and 1.95U/ml, respectively. Two-dimensional gel protein maps resulted in the separation and quantitative expression of different proteins by the microbial isolates. MALDI-TOF analysis and database search showed that the expressed proteins in this study closely relate to different glycoside hydrolases produced by other microbial species with an acceptable confidence level of 100%.

Keywords: cellulases, energy, two-dimensional gel electrophoresis, matrix-assisted laser desorption ionisation-time of flight, MALDI-TOF MS

Procedia PDF Downloads 125

Authors: Timea I. Hajnal-Jafari, Simonida S. Đurić, Dragana R. Stamenov

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Microbial inoculants from the group of symbiotic-nitrogen-fixing rhizobia are well known and widely used in production of legumes. On the other hand, nonsymbiotic plant growth promoting rhizobacteria (PGPR) are not commonly used in practice. The objective of this study was to examine the effects of soybean inoculation with symbiotic and nonsymbiotic bacteria on plant growth and seed yield of soybean. Microbiological activity in rhizospheric soil was also determined. The experiment was set up using a randomized block system in filed conditions with the following treatments: control-no inoculation; treatment 1-Bradyrhizobium japonicum; treatment 2-Azotobacter sp.; treatment 3-Bacillus sp..In the flowering stage of growth (FS) the number of nodules per plant (NPP), root length (RL), plant height (PH) and weight (PW) were measured. The number of pod per plant (PPP), number of seeds per pod (SPP) and seed weight per plant (SWP) were recorded at the end of vegetation period (EV). Microbiological analyses of soil included the determination of total number of bacteria (TNB), number of fungi (FNG), actinomycetes (ACT) and azotobacters (AZB) as well as the activity of the dehydrogenase enzyme (DHA). The results showed that bacterial inoculation led to the formation of root nodules regardless of the treatments with statistically no significant difference. Strong nodulation was also present in control treatment. RL and PH were positively influenced by inoculation with Azotobacter sp. and Bacillus sp., respectively. Statistical analyses of the number of PPP, SPP, and SWP showed no significant differences among investigated treatments. High average number of microorganisms were determined in all treatments. Most abundant were TNB (log No 8,010) and ACT (log No 6,055) than FNG and AZB with log No 4,867 and log No 4,025, respectively. The highest DHA activity was measured in the FS of soybean in treatment 3. The application of nonsymbiotic bacteria in soybean production can alleviate initial plant growth and help the plant to better overcome different stress conditions caused by abiotic and biotic factors.

Keywords: bacteria, inoculation, soybean, microbial activity

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Authors: Sara Arafeh, Kovriguine Evguine

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Cytochromes P450 are enzymes that require a supply of electrons to catalyze the synthesis of steroid hormones, fatty acids, and prostaglandin hormone. Cytochrome P450 Oxidoreductase (CYPOR), a membrane bound enzyme, provides these electrons in its open conformation. CYPOR has two cytosolic domains (FAD domain and FMN domain) and an N-terminal in the membrane. In its open conformation, electrons flow from NADPH, FAD, and finally to FMN where cytochrome P450 picks up these electrons. In the closed conformation, cytochrome P450 does not bind to the FMN domain to take the electrons. It was found that when the cytosolic domains are isolated, CYPOR could not bind to cytochrome P450. This suggested that the membrane environment is important for CYPOR function. This project takes the initiative to better understand the dynamics of CYPOR in its full length. Here, we determine the distance between specific sites in the FAD and FMN binding domains in CYPOR by Forster Resonance Energy Transfer (FRET) and Ultrafast TA spectroscopy with and without NADPH. The approach to determine these distances will rely on labeling these sites with red and infrared fluorophores. Mimic membrane attachment is done by inserting CYPOR in lipid nanodiscs. By determining the distances between the donor-acceptor sites in these domains, we can observe the open/closed conformations upon reducing CYPOR in the presence and absence of cytochrome P450. Such study is important to better understand CYPOR mechanism of action in various endosomal membranes including hepatic CYPOR which is vital in plasma cholesterol homeostasis. By investigating the conformational cycles of CYPOR, we can synthesize drugs that would be more efficient in affecting the steroid hormonal levels and metabolism of toxins catalyzed by Cytochrome P450.

Keywords: conformational cycle of CYPOR, cytochrome P450, cytochrome P450 oxidoreductase, FAD domain, FMN domain, FRET, Ultrafast TA Spectroscopy

Procedia PDF Downloads 275

Authors: Agatha Swasti Ayuning Tyas

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Hyperglycemia in Diabetes Mellitus (DM) is an important factor in cellular and vascular damage, which is caused by activation of C Protein Kinase, polyol and hexosamine track, and production of Advanced Glycation End-Products (AGE). Those mentioned before causes the accumulation of Reactive Oxygen Species (ROS). Oxidative stress increases the expression of proinflammatory factors IL-6 as one of many signs of endothelial disfunction. Genistein in soy milk has a high immunomodulator potential. Goat milk contains amino acids which have antioxidative potential. Fermented kefir has an anti-inflammatory activity which believed will also contribute in potentiating goat milk and soy milk. This study is a quasi-experimental posttest-only research to 30 Wistar mice. This study compared the levels of IL-6 between healthy Wistar mice group (G1) and 4 DM Wistar mice with intervention and grouped as follows: mice without treatment (G2), mice treated with 100% goat milk kefir (G3), mice treated with combination of 50% goat milk kefir and 50% soy milk kefir (G4), and mice treated with 100% soy milk kefir (G5). DM animal models were induced with Streptozotocin & Nicotinamide to achieve hyperglycemic condition. Goat milk kefir and soy milk kefir are given at a dose of 2 mL/kg body weight/day for four weeks to intervention groups. Blood glucose was analyzed by the GOD-POD principle. IL-6 was analyzed by enzyme-linked sandwich ELISA. The level of IL-6 in DM untreated control group (G2) showed a significant difference from the group treated with the combination of 50% goat milk kefir and 50% soy milk kefir (G3) (p=0,006) and the group treated with 100% soy milk kefir (G5) (p=0,009). Whereas the difference of IL-6 in group treated with 100% goat milk kefir (G3) was not significant (p=0,131). There is also synergism between glucose level and IL-6 in intervention groups treated with combination of 50% goat milk kefir and 50% soy milk kefir (G3) and the group treated with 100% soy milk kefir (G5). Combination of 50 % goat milk kefir and 50% soy milk kefir and administration of 100% soy milk kefir alone can control the level of IL-6 remained low in DM Wistar mice induced with streptozocin and nicotinamide.

Keywords: diabetes mellitus, goat milk kefir, soy milk kefir, interleukin 6

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Authors: N. Slepickova Kasalkova, P. Slepicka, L. Bacakova, V. Svorcik

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Tissue engineering includes combination of materials and techniques used for the improvement, repair or replacement of the tissue. Scaffolds, permanent or temporally material, are used as support for the creation of the "new cell structures". For this important component (scaffold), a variety of materials can be used. The advantage of some polymeric materials is their cytocompatibility and possibility of biodegradation. Poly(L-lactic acid) (PLLA) is a biodegradable,  semi-crystalline thermoplastic polymer. PLLA can be fully degraded into H₂O and CO₂. In this experiment, the effect of the surface modification of biodegradable polymer (performed by plasma treatment) on the various cell types was studied. The surface parameters and changes of the physicochemical properties of modified PLLA substrates were studied by different methods. Surface wettability was determined by goniometry, surface morphology and roughness study were performed with atomic force microscopy and chemical composition was determined using photoelectron spectroscopy. The physicochemical properties were studied in relation to cytocompatibility of human osteoblast (MG 63 cells), rat vascular smooth muscle cells (VSMC), and human stem cells (ASC) of the adipose tissue in vitro. A fluorescence microscopy was chosen to study and compare cell-material interaction. Important parameters of the cytocompatibility like adhesion, proliferation, viability, shape, spreading of the cells were evaluated. It was found that the modification leads to the change of the surface wettability depending on the time of modification. Short time of exposition (10-120 s) can reduce the wettability of the aged samples, exposition longer than 150 s causes to increase of contact angle of the aged PLLA. The surface morphology is significantly influenced by duration of modification, too. The plasma treatment involves the formation of the crystallites, whose number increases with increasing time of modification. On the basis of physicochemical properties evaluation, the cells were cultivated on the selected samples. Cell-material interactions are strongly affected by material chemical structure and surface morphology. It was proved that the plasma treatment of PLLA has a positive effect on the adhesion, spreading, homogeneity of distribution and viability of all cultivated cells. This effect was even more apparent for the VSMCs and ASCs which homogeneously covered almost the whole surface of the substrate after 7 days of cultivation. The viability of these cells was high (more than 98% for VSMCs, 89-96% for ASCs). This experiment is one part of the basic research, which aims to easily create scaffolds for tissue engineering with subsequent use of stem cells and their subsequent "reorientation" towards the bone cells or smooth muscle cells.

Keywords: poly(L-lactic acid), plasma treatment, surface characterization, cytocompatibility, human osteoblast, rat vascular smooth muscle cells, human stem cells

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Authors: Emine Teke, Soner Kuşlu, Sabri Çolak, Turan Çalban

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The aim of the present study was to investigate the dissolution kinetics of ulexite in sodium dihydrogen phosphate in a mechanical agitation system and also to declare an alternative reactant to produce the boric acid. Reaction temperature, concentration of sodium dihydrogen phosphate, stirring speed, solid-liquid ratio, and ulexite particle size were selected as parameters. The experimental results were successfully correlated by using linear regression and a statistical program. Dissolution curves were evaluated in order to test the shrinking core models for solid-fluid systems. It was observed that increase in the reaction temperature and decrease in the solid/liquid ratio causes an increase in the dissolution rate of ulexite. The activation energy was found to be 36.4 kJ/mol. The leaching of ulexite was controlled by diffusion through the ash (or product) layer.

Keywords: ulexite, sodium dihydrogen phosphate, leaching kinetics, boron

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Authors: B. Sadepovich Maikanov, Z. Shabanbayevich Adilbekov, R. Husainovna Mustafina, L. Tyulegenovna Auteleyeva

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This study involves detailed information about contaminants of honey in the Republic of Kazakhstan. The requirements of the technical regulation ‘Requirements to safety of honey and bee products’ and GOST 19792-2001 were taken into account in this research. Contamination of honey by antibiotics wqs determined by the IEA (immune-enzyme analysis), Ridder analyzer and Tecna produced test systems. Voltammetry (TaLab device) was used to define contamination by salts of heavy metals and gamma-beta spectrometry, ‘Progress BG’ system, with preliminary ashing of the sample of honey was used to define radioactive contamination. This article pointed out that residues of chloramphenicol were detected in 24% of investigated products, in 22% of them –streptomycin, in 7.3% - sulfanilamide, in 2.4% - tylosin, and in 12% - combined contamination was noted. Geographically, the greatest degree of contamination of honey with antibiotics occurs in the Northern Kazakhstan – 54.4%, and Southern Kazakhstan - 50%, and the lowest in Central and Eastern Kazakhstan with 30% and 25%, respectively. Generally, pollution by heavy metals is within acceptable limits, but the contamination from lead is highest in the Akmola region. The level of radioactive cesium and strontium is also within acceptable concentrations. The highest radioactivity in terms of cesium was observed in the East Kazakhstan region - 49.00±10 Bq/kg, in Akmola, North Kazakhstan and Almaty - 12.00±5, 11.05±3 and 19.0±8 Bq/kg, respectively, while the norm is 100 Bq/kg. In terms of strontium, the radioactivity in the East Kazakhstan region is 25.03±15 Bq/kg, while in Akmola, North Kazakhstan and Almaty regions it is 12.00±3, 10.2±4 and 1.0±2 Bq/kg, respectively, with the norm of 80 Bq/kg. This accumulation is mainly associated with the environmental degradation, feeding and treating of bees. Moreover, in the process of collecting nectar, external substances can penetrate honey. Overall, this research determines factors and reasons of honey contamination.

Keywords: antibiotics, contamination of honey, honey, radionuclides

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Authors: Rania Samir Zaki

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Because of their inhibitory effects, selected probiotic Lactobacilli may be used as antimicrobial against some hazardous microorganisms responsible for spoilage of fresh minced beef (cattle) minced meat and camel minced meat. Lactic acid bacteria were isolated from camel meat. These included 10 isolates; 1 Lactobacillus fermenti, 4 Lactobacillus plantarum, 4 Lactobacillus pulgaricus, 3 Lactobacillus acidophilus and 1 Lactobacillus brevis. The most efficient inhibitory organism was Lactobacillus plantarum which can be used as a propiotic with antibacterial activity. All microbiological analyses were made at the time 0, first day and the second day at altered ranges of temperature [4±2 ⁰C (chilling temperature), 25±2 ⁰C, and 38±2 ⁰C]. Results showed a significant decrease of pH 6.2 to 5.1 within variant types of meat, in addition to reduction of Total Bacterial Count, Enterococci, Bacillus cereus and Escherichia coli together with the stability of Coliforms and absence of Staphylococcus aureus.

Keywords: antibacterial, camel meat, inhibition, probiotics

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Authors: Lotfi Hedi Khezami, Mohamed Ben Rabha, N. Sboui, Mounir Gaidi, B. Bessais

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Metal-nano particle-assisted Chemical Etching is an extraordinary developed wet etching method of producing uniform semiconductor nano structure (nano wires) from patterned metallic film on crystalline silicon surface. The metal films facilitate the etching in HF and H2O2 solution and produce silicon nanowires (SiNWs). Creation of different SiNWs morphologies by changing the etching time and its effects on optical and opto electronic properties was investigated. Combination effect of formed SiNWs and stain etching treatment in acid (HF/HNO3/H2O) solution on the surface morphology of Si wafers as well as on the optical and opto electronic properties are presented in this paper.

Keywords: stain etching, porous silicon, silicon nanowires, reflectivity, lifetime, solar cells

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Authors: Ravisinh Solanki, Ravi Patel, Chhaganbhai Patel

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Purpose: The purpose of this study is to develop a methodology for the risk assessment and evaluation of nitrosamine impurities in Glipizide antidiabetic formulations. Nitroso compounds, including nitrosamines, have emerged as significant concerns in drug products, as highlighted by the ICH M7 guidelines. This study aims to identify known and potential sources of nitrosamine impurities that may contaminate Glipizide formulations and assess their presence. By determining observed or predicted levels of these impurities and comparing them with regulatory guidance, this research will contribute to ensuring the safety and quality of combination antidiabetic drug products on the market. Factors contributing to the presence of genotoxic nitrosamine contaminants in glipizide medications, such as secondary and tertiary amines, and nitroso group-complex forming molecules, will be investigated. Additionally, conditions necessary for nitrosamine formation, including the presence of nitrosating agents, and acidic environments, will be examined to enhance understanding and mitigation strategies. Method: The methodology for the study involves the implementation of the N-Nitroso Acid Precursor (NAP) test, as recommended by the WHO in 1978 and detailed in the 1980 International Agency for Research on Cancer monograph. Individual glass vials containing equivalent to 10mM quantities of Glipizide is prepared. These compounds are dissolved in an acidic environment and supplemented with 40 mM NaNO2. The resulting solutions are maintained at a temperature of 37°C for a duration of 4 hours. For the analysis of the samples, an HPLC method is employed for fit-for-purpose separation. LC resolution is achieved using a step gradient on an Agilent Eclipse Plus C18 column (4.6 X 100 mm, 3.5µ). Mobile phases A and B consist of 0.1% v/v formic acid in water and acetonitrile, respectively, following a gradient mode program. The flow rate is set at 0.6 mL/min, and the column compartment temperature is maintained at 35°C. Detection is performed using a PDA detector within the wavelength range of 190-400 nm. To determine the exact mass of formed nitrosamine drug substance related impurities (NDSRIs), the HPLC method is transferred to LC-TQ-MS/MS with the same mobile phase composition and gradient program. The injection volume is set at 5 µL, and MS analysis is conducted in Electrospray Ionization (ESI) mode within the mass range of 100−1000 Daltons. Results: The samples of NAP test were prepared according to the protocol. The samples were analyzed using HPLC and LC-TQ-MS/MS identify possible NDSRIs generated in different formulations of glipizide. It was found that the NAP test generated a various NDSRIs. The new finding, which has not been reported yet, discovered contamination of Glipizide. These NDSRIs are categorised based on the predicted carcinogenic potency and recommended its acceptable intact in medicines. The analytical method was found specific and reproducible.

Keywords: NDSRI, nitrosamine impurities, antidiabetic, glipizide, LC-MS/MS

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Authors: Kwangman Park, Jinhee Kang, Suhee Kim, Dohyeon Yu, Kyoungseong Choi, Jinho Park

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Statement of the Problem: Diarrhea is a major cause of death in young calves. This causes great economic damage to the livestock industry. These diarrhea cause dehydration, decrease blood flow, lower the pH and degrade enzyme function. In the past, serum screening was not possible in the field. However, now with the spread of portable serum testing devices, it is now possible to conduct tests directly on field. Thus, accurate serological changes can be identified and used in the field of large animals. Methodology and Theoretical Orientation: The test groups were calves from 1 to 44 days old. The status of the feces was divided into four grade to determine the severity of diarrhea (grade 0,1,2,3). Grade 0, 1 is considered to have no diarrhea. Grade 2, 3 is considered to diarrhea positive group. One or more viruses were detected in this group. Diarrhea negasitive group consisted of 57 calves (Asan=30, Samrye=27). Diarrhea positive group consisted of 34 calves (Kimje=27, Geochang=7). The feces of all calves were analyzed by PCR Test. Blood sample was measured using an automatic blood analyzer(i-STAT, Abbott inc. Illinois, US). Calves were divided into 3 groups according to age. Group 1 is 1 to 14 days old. Group 2 is 15 to 28 days old. Group 3 is more than 28 days old. Findings: Diarrhea caused an increase in HCT due to dehydration. The difference from normal was highest in 15 to 28 days old (p < 0.01). At all ages, bicarbonate decreased compared to normal, and therefore pH decreased. Similar to HCT, the largest difference was observed between 15 and 28 days (p < 0.01). The pCO₂ decreases to compensate for the decrease in pH. Conclusion and Significance: At all ages, HCT increases, and bicarbonate, pH, and pCO₂ decrease in diarrhea calves. The calf from 15 days to 28 days shows the most difference from normal. Over 28 days of age, weight gain and homeostasis ability increase, diarrhea is seen in the stool, there are fewer hematologic changes than groups below 28 days of age.

Keywords: calves, diarrhea, hematological changes, i-STAT

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Authors: Nada Bezić, Valerija Dunkić, Rušćić Mirko

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Astragalus monspessulanus L. ssp. illyricus (Bernhardt) Chater is endemic species of Fabaceae family and belongs to hemicryptophyte. This plant grows wild in the sub-Mediterranean area. We analyzed the composition of the essential oil of stems and leaves of A. monspessulanus L. ssp. Illyricus, collected in Tijarica, near Split, Croatia. Water distilled essential oils from aerial parts of investigation plant have been analysed by GC and GC/MS using VF-5ms capillary column. The total yield of oil was 0.08%, based on dry weight of samples. Thirty-eight compounds were representing 88.5% of the total oil. This essential oil was characterized by a high concentration of cis-myrtanol (20.5%), geranyl acetate (9.6%) and phytone (6.6%). Previous research in the species A. monspessulanus have included flavoalkaloids and flavonoids composition. The present study gives additional knowledge about secondary metabolites contents on the genus Astragalus.

Keywords: essential oil, isovaleric acid, Valeriana tuberosa, geranyl acetate, phytone

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Authors: Vassilena Dakova, Mariana Panayotova-Pencheva

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The present work focused on the study of pulmonary helminth-fauna of the stone marten in Bulgaria in terms of which the data are little. For the purpose, four stone martens were helminthologically necropsied according to the common technique. In addition, some of the injured lung parts were investigated after their boiling in lactic acid and subsequent compression. Four nematode species from different families of order Strongylida and Trichocephalida were found in the lungs. These were Crenosoma petrowi Morosov, 1939; Eucoleus aerophilus Creplin, 1839; Filaroides martis Werner, 1782 and Sobolevingylus petrowi Romanov, 1952. Some of the parasite structures with taxonomic importance were measured and described. According to our best knowledge, the species F. martis and S. petrowi are recorded for the first time as a part of the helminth-fauna of Southeast Europe and Bulgaria in particular.

Keywords: Bulgaria, Crenosoma petrowi, Eucoleus aerophilus, Filaroides martis, lung parasites, Sobolevingylus petrowi, stone martens

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Authors: Simona Moravcova, Jiri Novotny, Zdenka Bendova

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The pineal gland of the vertebrate brain is a circumventricular organ which serves as a major neuroendocrine gland with the primary function of rhythmic secretion of neurohormone melatonin under the control of the hypothalamic suprachiasmatic nucleus (SCN). Soon after the onset of the darkness, the activity of the key rate-limiting enzyme for melatonin synthesis, arylalkylamine N-acetyltransferase (AANAT), raises due to the increased release of norepinephrine from sympathetic neurons terminating on the parenchymal cells where it binds to β-adrenergic receptors. Melatonin codes the length of the night, and it is well recognized for its anti-inflammatory effects. However, to our knowledge, less is known about the effect of the immune system on the melatonin biosynthesis and the precise role of the STAT3 in the signaling pathway leading to the expression of AANAT. Lipopolysaccharide (LPS) is the essential component in the outer surface membrane of gram-negative bacteria and acts as a strong stimulator of natural and innate immunity. STAT3 acts as an important factor in immune response. Here we investigated the effect of LPS on the components of the melatonergic synthetic pathway in the pineal gland. The experiments were performed both in vivo and in vitro. The changes in AANAT activity were determined by radioenzymatic assay. PCR analyses were carried out to detect aa-nat, icer, spi-3 and stat3 gene expression. From our results, it is apparent that the high basal level of phosphorylated forms of STAT3 can be elevated after systemic as well as in vitro administration of LPS. Our experiments have shown that LPS reduces melatonin synthesis, nevertheless, the activity of AANAT was increased. Moreover, the basal level of phosphorylated STAT3 counteracts β-adrenergic receptor-mediated aa-nat gene expression and sustains its own and spi-3 gene expression. In conclusion, LPS can affect immunomodulators such as melatonin in the pineal gland.

Keywords: AANAT, lipopolysaccharide, pineal gland, rat, STAT3

Procedia PDF Downloads 168

Authors: M. Kouyate, M. Sangare, S. Samake, S. Keita, H. G. Kim, D. H. Geschwind

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Background & Objectives: Human genetic studies can be expensive, even unaffordable, in developing countries, partly due to the sequencing costs. Our aim is to pilot the use of bioinformatics tools to guide scientifically valid, locally relevant, and economically sound autism genetic research in Mali. Methods: The following databases, NCBI, HGMD, and LSDB, were used to identify hot point mutations. Phenotype, transmission pattern, theoretical protein expression in the brain, the impact of the mutation on the 3D structure of the protein) were used to prioritize selected autism genes. We used the protein database, Modeller, and clustal W. Results: We found Mef2c (Gly27Ala/Leu38Gln), Pten (Thr131IIle), Prodh (Leu289Met), Nme1 (Ser120Gly), and Dhcr7 (Pro227Thr/Glu224Lys). These mutations were associated with endonucleases BseRI, NspI, PfrJS2IV, BspGI, BsaBI, and SpoDI, respectively. Gly27Ala/Leu38Gln mutations impacted the 3D structure of the Mef2c protein. Mef2c protein sequences across species showed a high percentage of similarity with a highly conserved MADS domain. Discussion: Mef2c, Pten, Prodh, Nme1, and Dhcr 7 gene mutation frequencies in the Malian population will be very informative. PCR coupled with restriction enzyme digestion can be used to screen the targeted gene mutations. Sanger sequencing will be used for confirmation only. This will cut down considerably the sequencing cost for gene-to-gene mutation screening. The knowledge of the 3D structure and potential impact of the mutations on Mef2c protein informed the protein family and altered function (ex. Leu38Gln). Conclusion & Future Work: Bio-informatics will positively impact autism research in Mali. Our approach can be applied to another neuropsychiatric disorder.

Keywords: bioinformatics, endonucleases, autism, Sanger sequencing, point mutations

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Authors: Leo Paul Vaurs, Sonia Heaven, Charles Banks

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Municipal solid waste (MSW) is a virtually unlimited source of lignocellulosic material in the form of a waste paper/cardboard mixture which can be converted into fermentable sugars via cellulolytic enzyme hydrolysis in a biorefinery. The extraction of the lignocellulosic fraction and its preparation, however, are energy and water demanding processes. The waste water generated is a rich organic liquor with a high Chemical Oxygen Demand that can be partially cleaned while generating biogas in an Upflow Anaerobic Sludge Blanket bioreactor and be further re-used in the process. In this work, an experiment was designed to determine the critical contaminant concentrations in water affecting either anaerobic digestion or enzymatic hydrolysis by simulating multiple water re-circulations. It was found that re-using more than 16.5 times the same water could decrease the hydrolysis yield by up to 65 % and led to a complete granules desegregation. Due to the complexity of the water stream, the contaminant(s) responsible for the performance decrease could not be identified but it was suspected to be caused by sodium, potassium, lipid accumulation for the anaerobic digestion (AD) process and heavy metal build-up for enzymatic hydrolysis. The experimental data were incorporated into a Water Pinch technology based model that was used to optimize the water re-utilization in the modelled system to reduce fresh water requirement and wastewater generation while ensuring all processes performed at optimal level. Multiple scenarios were modelled in which sub-process requirements were evaluated in term of importance, operational costs and impact on the CAPEX. The best compromise between water usage, AD and enzymatic hydrolysis yield was determined for each assumed contaminant degradations by anaerobic granules. Results from the model will be used to build the first MSW based biorefinery in the USA.

Keywords: anaerobic digestion, enzymatic hydrolysis, municipal solid waste, water optimization

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Authors: Hinda Berghiche, Hamida Benradia, Noureddine Soltani

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Pesticides are widely used in crop production and are known to induce a major contamination of ecosystems especially in aquatic environments. The leaching of a large amount of pollutants derived from agricultural activities (fertilizers, pesticides) might contaminate rivers which diverse into the likes and estuarine and coastal environments affecting several organisms such as crustacean species. In this context, there is searched for new selective insecticides with minimal toxic effects on the environment and human health such as growth insect regulators (GIRs). The current study aimed to examine the impact of novaluron (CE 20%), a potent benzoylphenylurea derivative insecticide on mosquito larvae, against non-target shrimp, Palaemon adspersus (Decapoda, Palaemonidae). The compound was tested at two concentrations (0.91 mg/L and 4.30 mg/L) corresponding respectively to the LC50 and LC90 determined against fourth-instar larvae of Culiseta longiareolata (Diptera, Culicidae). The molting hormone titer was determined in the haemolymph by an enzyme-immunoassay, while chitin was measured in peripheral integument at different stages during the molting cycle. Under normal conditions, the haemolymphatic ecdysteroid concentrations increased during the molting cycle to reach peak at stage D. In the treated series, we note absence of the peak at stage D and an increase at stages B, C and D as compared to the controls. Concerning the chitin amounts, we observe an increase from stage A to stage C followed by a decrease at stage D. Exposition of shrimps to novaluron resulted in a significant decrease of values at all molting stages with a dose-response effect. Thus, the insecticide can present secondary effects on this non-target arthropod species.

Keywords: toxicology, novaluron, crustacean, palaemon adspersus, ecdysteroids, cuticle, chitin

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Authors: Khaled Alshammari

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Ciprofloxacin (CIP) is a common antibiotic drug used as a strudy electron donor that interacts with dynamic π -acceptors such as 2,3-dinitrosalsylic acid (HDNS) and Tetracyanoethylene (TCNE) for synthesizing a new model of charge transfer (CT) complexes. The synthesized complexes were identified using diverse analytical methods such as UV–vis spectra, photometric titration measurements, FT-IR, HNMR Spectroscopy, and thermogravimetric analysis techniques (TGA/DTA). The stoichiometries for all the formed complexes were found to be a 1:1 M ratio between the reactants. The characteristic spectroscopic properties such as transition dipole moment (µ), oscillator strength (f), formation constant (KCT), ionization potential (ID), standard free energy (∆G), and energy of interaction (ECT) for the CT-complexes were collected. The developed CT complexes were tested for their toxicity on main organs, antimicrobial activity, antioxidant activity, and biofilm formation.

Keywords: biological, biofilm, toxicity, thermal analysis, charge transfer, spectroscopy

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Authors: Medina Kadiri

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Blue-green otherwise called cyanobacteria, produce an array of biotoxins grouped into five categories notably hapatotoxins, neurotoxins, cytotoxins, dermatotoxins, and irritant toxins. Microcystins which are examples of hepatotoxins produced by blue-green algae Microcystins comprise the most common group of the cyanobacterial toxins. Blue-green algae flourish in aquatic environments, whether marine, brackish or freshwater, producing blooms in different forms such as microscopic, mats, or unsightly odoriferous scums. Microcystins biotoxins cause a plethora of animal and human hazards such as liver damage/cirrhosis and cancer, kidney damage, dermatitis, tinnitus, gastroenteritis, sore throat, nausea, myalgia, neurological problems, respiratory irritation and death. Water samples were collected from coastal regions of Nigeria in March 2014, June 2014, October 2014 and January 2015 and analyzed with Enzyme Linked Immunosorbent Assay (ELISA) kits. Microcystin biotoxin was recorded in all sites both during dry and wet seasons. The range of microcystins found was 0.000041-There was a seasonal trend of increasing microcystin concentrations from March till Octobers and a decrease thereafter. Generally in the oceanic waters, microcystin levels were highest at Cross Rivers in March and January, Barbeach in June and Lekki in October. In the adjoining riverine ecosystems, on the other hand, the highest concentrations of microcystin were observed at Akwa Ibom in March, June and October and in Bayelsa in January. Continuous monitoring and screening of coastal water bodies is suggested to minimize the health risks of cyanobacterial biotoxins to coastal communities of Nigeria.

Keywords: biotoxins, harmful algae, marine, microcystin, Nigeria

Procedia PDF Downloads 278