Search results for: methanol to olefins

Authors: Amanda C. Q. M. Vieira, Cybelly M. Melo, Camila B. M. Figueirêdo, Giovanna C. R. M. Schver, Salvana P. M. Costa, Magaly A. M. de Lyra, Ping I. Lee, José L. Soares-Sobrinho, Pedro J. Rolim-Neto, Mônica F. R. Soares

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Epiisopiloturine (EPI) is a drug candidate that is extracted from Pilocarpus microphyllus and isolated from the waste of Pilocarpine. EPI has demonstrated promising schistosomicidal, leishmanicide, anti-inflammatory and antinociceptive activities, according to in vitro studies that have been carried out since 2009. However, this molecule shows poor aqueous solubility, which represents a problem for the release of the drug candidate and its absorption by the organism. The purpose of the present study is to investigate the extent of enhancement of kinetic solubility of a solid solution (SS) of EPI in hipromellose phthalate HP-55 (HPMCP), an enteric polymer carrier. SS was obtained by the solvent evaporation methodology, using acetone/methanol (60:40) as solvent system. Both EPI and polymer (drug loading 10%) were dissolved in this solvent until a clear solution was obtained, and then dried in oven at 60ºC during 12 hours, followed by drying in a vacuum oven for 4 h. The results show a considerable modification in the crystalline structure of the drug candidate. For instance, X-ray diffraction (XRD) shows a crystalline behavior for the EPI, which becomes amorphous for the SS. Polarized light microscopy, a more sensitive technique than XRD, also shows completely absence of crystals in SS sample. Differential Scanning Calorimetric (DSC) curves show no signal of EPI melting point in SS curve, indicating, once more, no presence of crystal in this system. Interaction between the drug candidate and the polymer were found in Infrared microscopy, which shows a carbonyl 43.3 cm-1 band shift, indicating a moderate-strong interaction between them, probably one of the reasons to the SS formation. Under sink conditions (pH 6.8), EPI SS had its dissolution performance increased in 2.8 times when compared with the isolated drug candidate. EPI SS sample provided a release of more than 95% of the drug candidate in 15 min, whereas only 45% of EPI (alone) could be dissolved in 15 min and 70% in 90 min. Thus, HPMCP demonstrates to have a good potential to enhance the kinetic solubility profile of EPI. Future studies to evaluate the stability of SS are required to conclude the benefits of this system.

Keywords: epiisopiloturine, hipromellose phthalate HP-55, pharmaceuticaltechnology, solubility

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Authors: Abhay Asthana, Shally Toshkhani, Gyati Shilakari

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The objective of the present research work is to develop a topical biodegradable dermal patch based formulation to aid accelerated wound healing. It is always better for patient compliance to be able to reduce the frequency of dressings with improved drug delivery and overall therapeutic efficacy. In present study optimized formulation using biodegradable components was obtained evaluating polymers and excipients (HPMC K4M, Ethylcellulose, Povidone, Polyethylene glycol and Gelatin) to impart significant folding endurance, elasticity, and strength. Molten gelatin was used to get a mixture using ethylene glycol. Chitosan dissolved in acidic medium was mixed with stirring to Gelatin mixture. With continued stirring to the mixture Curcumin was added with the aid of DCM and Methanol in an optimized ratio of 60:40 to get homogenous dispersion. Polymers were dispersed with stirring in the final formulation. The mixture was sonicated casted to get the film form. All steps were carried out under strict aseptic conditions. The final formulation was a thin uniformly smooth textured film with dark brown-yellow color. The film was found to have folding endurance was around 20 to 21 times without a crack in an optimized formulation at RT (23°C). The drug content was in range 96 to 102% and it passed the content uniform test. The final moisture content of the optimized formulation film was NMT 9.0%. The films passed stability study conducted at refrigerated conditions (4±0.2°C) and at room temperature (23 ± 2°C) for 30 days. Further, the drug content and texture remained undisturbed with stability study conducted at RT 23±2°C for 45 and 90 days. Percentage cumulative drug release was found to be 80% in 12h and matched the biodegradation rate as tested in vivo with correlation factor R2>0.9. In in vivo study administration of one dose in equivalent quantity per 2 days was applied topically. The data demonstrated a significant improvement with percentage wound contraction in contrast to control and plain drug respectively in given period. The film based formulation developed shows promising results in terms of stability and in vivo performance.

Keywords: wound healing, biodegradable, polymers, patch

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Authors: Kathryn Nderitu, Atunga Nyachieo, Ezekiel Mecha

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Introduction: Solanum nigrum , also known as black nightshade, biosynthesizes various phytochemical compounds with various pharmacological activities, including treating cardiovascular diseases and type 2 diabetes, among others. Materials and Methods: To assess the anti-obesity effects of Solanum nigrum using high-fat-fed diet rats, Sprague Dawley male rats (n = 35) of weights 160–180 g were assigned randomly into seven groups comprising n = 5 rats each. Each group was fed for 11 weeks as follows: normal group (normal chow rat feed); high-fat diet control (HFD); HFD and standard drug (Orlistat 30 mg/kg bw); HFD and methanolic extract 150 mg/kgbw; HFD and methanolic extract 300 mg/kgbw; HFD and dichloromethane extract 150 mg/kgbw; HFD and dichloromethane extract 300 mg/kgbw. Body mass index and food intake were monitored per week, and an oral glucose tolerance test was measured in weeks 5 and 10. Lipid profiles, liver function tests, adipose tissue, liver weights, and phytochemical analysis of Solanum nigrum were later carried out. Results: High-fat diet control group rats exhibited a significant increase in body mass index (BMI), while rats administered with leaf extracts of Solanum nigrum showed a reduction in BMI. Both low doses of dichloromethane (150 mg/kgbw) and high doses of methanol extracts (300 mg/kgbw) showed a better reduction in BMI than the other treatment groups. A significant decrease (p <0.05) in low-density lipoprotein-cholesterol, triglycerides, and cholesterol was observed among the rats administered with Solanum nigrum extracts compared to those of HFD control. Moreover, the HFD control group significantly increased liver and adipose tissue weights compared to other treatment groups (p<0.05). Solanum nigrum also decreased glycemic levels and normalized the hepatic enzymes of HFD control. However, food intake among the groups showed no significant difference (p>0.05). Qualitative analysis of Solanum nigrum leaf extracts indicated the presence of various bioactive compounds associated with anti-obesity. Conclusion: These results validate the use of Solanum nigrum in controlling obesity.

Keywords: solanum nigrum, High fat diet, phytocompounds, obesity

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Authors: Wighens Ngoie Ilunga, Pamela J. Welz, Olewaseun O. Oyekola, Daniel Ikhu-Omoregbe

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Currently, most sludge from the wastewater treatment plants of edible oil factories is disposed to landfills, but landfill sites are finite and potential sources of environmental pollution. Production of biodiesel from wastewater sludge can contribute to energy production and waste minimization. However, conventional biodiesel production is energy and waste intensive. Generally, biodiesel is produced from the transesterification reaction of oils with alcohol (i.e., Methanol, ethanol) in the presence of a catalyst. Homogeneously catalysed transesterification is the conventional approach for large-scale production of biodiesel as reaction times are relatively short. Nevertheless, homogenous catalysis presents several challenges such as high probability of soap. The current study aimed to reuse wastewater sludge from the edible oil industry as a novel feedstock for both monounsaturated fats and bioethanol for the production of biodiesel. Preliminary results have shown that the fatty acid profile of the oilseed wastewater sludge is favourable for biodiesel production with 48% (w/w) monounsaturated fats and that the residue left after the extraction of fats from the sludge contains sufficient fermentable sugars after steam explosion followed by an enzymatic hydrolysis for the successful production of bioethanol [29% (w/w)] using a commercial strain of Saccharomyces cerevisiae. A novel nano-magnetic catalyst was synthesised from mineral processing alkaline tailings, mainly containing dolomite originating from cupriferous ores using a modified sol-gel. The catalyst elemental chemical compositions and structural properties were characterised by X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infra-red (FTIR) and the BET for the surface area with 14.3 m²/g and 34.1 nm average pore diameter. The mass magnetization of the nano-magnetic catalyst was 170 emu/g. Both the catalytic properties and reusability of the catalyst were investigated. A maximum biodiesel yield of 78% was obtained, which dropped to 52% after the fourth transesterification reaction cycle. The proposed approach has the potential to reduce material costs, energy consumption and water usage associated with conventional biodiesel production technologies. It may also mitigate the impact of conventional biodiesel production on food and land security, while simultaneously reducing waste.

Keywords: biodiesel, bioethanol, edible oil wastewater sludge, nano-magnetism

Procedia PDF Downloads 130

Authors: Onyekachi Ogbonnaya Iroanya, Taiye Abdullahi Gegele, Frank Tochukwu Egwuatu

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Introduction: Forensic taphonomy is a multifaceted area that incorporates decomposition, chemical and biological cadaver exposure in post-mortem event chronology and reconstruction to predict the Post Mortem Interval (PMI). The aim of this study was to evaluate the integrity of DNA extracted from the remains of embalmed decomposed Sus domesticus tissues buried in different soil types. Method: A total of 12 limbs of Sus domesticus weighing between 0.7-1.4 kg were used. Each of the samples across the groups was treated with 10% formaldehyde, absolute methanol and 50% Pine oil for 24 hours before burial except the control samples, which were buried immediately. All samples were buried in shallow simulated Clay, Sandy and Loamy soil graves for 12 months. The DNA for each sample was extracted and quantified with Nanodrop Spectrophotometer (6305 JENWAY spectrometers). The rate of decomposition was examined through the modified qualitative decomposition analysis. Extracted DNA was amplified through PCR and bands visualized via gel electrophoresis. A biochemical enzyme assay was done for each burial grave soil. Result: The limbs in all burial groups had lost weight over the burial period. There was a significant increase in the soil urease level in the samples preserved in formaldehyde across the 3 soil type groups (p≤0.01). Also, the control grave soils recorded significantly higher alkaline phosphatase, dehydrogenase and calcium carbonate values compared to experimental grave soils (p≤0.01). The experimental samples showed a significant decrease in DNA concentration and purity when compared to the control groups (p≤0.01). Obtained findings of the soil biochemical analysis showed the embalming treatment altered the relationship between organic matter decomposition and soil biochemical properties as observed in the fluctuations that were recorded in the soil biochemical parameters. The PCR amplified DNA showed no bands on the gel electrophoresis plates. Conclusion: In criminal investigations, factors such as burial grave soil, grave soil biochemical properties, antemortem exposure to embalming chemicals should be considered in post-mortem interval (PMI) determination.

Keywords: forensic taphonomy, post-mortem interval (PMI), embalmment, decomposition, grave soil

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Authors: Hossein Rassi, Marjan Moradi Fard, Samaneh Niko

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Human papillomavirus type 16 and 18 are closely associated with the development of human cervical carcinoma, which is one of the most common causes of cancer death in women worldwide. At present, HPV type 18 accounts for about 34 % of all HPV infections in Iran and the most promising vaccine against HPV infection is based on the L1 major capsid protein. The L1 protein of HPV18 has the capacity to self-assemble into capsomers or virus-like particles (VLPs) that are non-infectious, highly immunogenic and allowing their use in vaccine production. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system used to produce high levels of heterologous proteins. In this study we expressed HPV18 L1 VLPs in P. pastoris. The gene encoding the major capsid protein L1 of the high-risk HPV type 18 was isolated from Iranian patient by PCR and inserted into pTG19-T vector to obtain the recombinant expression vector pTG19-HPV18-L1. Then, the pTG19-HPV18-L1 was transformed into E. coli strain DH5α and the recombinant protein HPV18 L1 was expressed under IPTG induction in soluble form. The HPV18 L1 gene was excised from recombinant plasmid with XhoI and EcoRI enzymes and ligated into the yeast expression vector pPICZα linearized with the same enzymes, and transformed into P. pastoris. Induction and expression of HPV18 L1 protein was demonstrated by BMGY/BMMY and RT PCR. The parameters for induced cultivation for strain in P. pastoris KM71 with HPV16L1 were investigated in shaking flask cultures. After induced cultivation BMMY (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 24 h at 37 degrees C for 96 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by P. pastoris for HPV-18 L1 gene. The VLP-based HPV vaccines can prevent persistent HPV18 infections and cervical cancer in Iran. The HPV-18 L1 gene was expressed successfully in E.coli, which provides necessary basis for preparing HPV-18 L1 vaccine in human. Also, HPV type 6 L1 proteins expressed in Pichia pastoris will facilitate the HPV vaccine development and structure-function study.

Keywords: Pichia pastoris, L1 virus-like particles, human papillomavirus type 18, biotechnology

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Authors: M. S. Abd El-Sadek, M. A. Omar, Gharib M. Taha

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In recent years, photocatalytic degradation of various kinds of organic and inorganic pollutants using semiconductor powders as photocatalysts has been extensively studied. Owing to its relatively high photocatalytic activity, biological and chemical stability, low cost, nonpoisonous and long stable life, Tin oxide materials have been widely used as catalysts in chemical reactions, including synthesis of vinyl ketone, oxidation of methanol and so on. Tin oxide (SnO₂), with a rutile-type crystalline structure, is an n-type wide band gap (3.6 eV) semiconductor that presents a proper combination of chemical, electronic and optical properties that make it advantageous in several applications. In the present work, SnO₂ nanoparticles were synthesized at room temperature by the sol-gel process and thermohydrolysis of SnCl₂ in isopropanol by controlling the crystallite size through calculations. The synthesized nanoparticles were identified by using XRD analysis, TEM, FT-IR, and Uv-Visible spectroscopic techniques. The crystalline structure and grain size of the synthesized samples were analyzed by X-Ray diffraction analysis (XRD) and the XRD patterns confirmed the presence of tetragonal phase SnO₂. In this study, Methylene blue degradation was tested by using SnO₂ nanoparticles (at different calculations temperatures) as a photocatalyst under sunlight as a source of irradiation. The results showed that the highest percentage of degradation of Methylene blue dye was obtained by using SnO₂ photocatalyst at calculations temperature 800 ᵒC. The operational parameters were investigated to be optimized to the best conditions which result in complete removal of organic pollutants from aqueous solution. It was found that the degradation of dyes depends on several parameters such as irradiation time, initial dye concentration, the dose of the catalyst and the presence of metals such as silver as a dopant and its concentration. Percent degradation was increased with irradiation time. The degradation efficiency decreased as the initial concentration of the dye increased. The degradation efficiency increased as the dose of the catalyst increased to a certain level and by further increasing the SnO₂ photocatalyst dose, the degradation efficiency is decreased. The best degradation efficiency on which obtained from pure SnO₂ compared with SnO₂ which doped by different percentage of Ag.

Keywords: SnO₂ nanoparticles, a sol-gel method, photocatalytic applications, methylene blue, degradation efficiency

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Authors: Abhay Asthana, Gyati Shilakari Asthana

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It’s the patient compliance and stability in combination with controlled drug delivery and biocompatibility that forms the core feature in present research and development of sustained biodegradable patch formulation intended for wound healing. The aim was to impart sustained degradation, sterile formulation, significant folding endurance, elasticity, biodegradability, bio-acceptability and strength. The optimized formulation was developed using component including polymers including Hydroxypropyl methyl cellulose, Ethylcellulose, and Gelatin, and Citric Acid PEG Citric acid (CPEGC) triblock dendrimers and active Curcumin. Polymeric mixture dissolved in geometric order in suitable medium through continuous stirring under ambient conditions. With continued stirring Curcumin was added with aid of DCM and Methanol in optimized ratio to get homogenous dispersion. The dispersion was sonicated with optimum frequency and for given time and later casted to form a patch form. All steps were carried out under under strict aseptic conditions. The formulations obtained in the acceptable working range were decided based on thickness, uniformity of drug content, smooth texture and flexibility and brittleness. The patch kept on stability using butter paper in sterile pack displayed folding endurance in range of 20 to 23 times without any evidence of crack in an optimized formulation at room temperature (RT) (24 ± 2°C). The patch displayed acceptable parameters after stability study conducted in refrigerated conditions (8±0.2°C) and at RT (24 ± 2°C) upto 90 days. Further, no significant changes were observed in critical parameters such as elasticity, biodegradability, drug release and drug content during stability study conducted at RT 24±2°C for 45 and 90 days. The drug content was in range 95 to 102%, moisture content didn’t exceeded 19.2% and patch passed the content uniformity test. Percentage cumulative drug release was found to be 80% in 12h and matched the biodegradation rate as drug release with correlation factor R2>0.9. The biodegradable patch based formulation developed shows promising results in terms of stability and release profiles.

Keywords: sustained biodegradation, wound healing, polymers, stability

Procedia PDF Downloads 325

Authors: Sunil Kumar Ghosh

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Som plant (Machilus bombycina King) is an important plant in agroforestry system. It is cultivated in north -east part of India. It is cultivated in agricultural land by the marginal farmers for multi-storeyed cultivation with intercropping. Localized cottage industries are involved with this plant like sericulture industry (muga silk worm cultivation). Clothes are produced from this sericulture industry. Leaves of som plants are major food of muga silk worm ( Antherea assama ). Nutritional value of leaves plays an important role in the larval growth and silk productivity. The plant also has timber value. The plant is susceptible to mite pest (Tetranychus sp.) causes heavy damage to tender leaves. Lower population was recorded during 7th to 38th standard week, during 3rd week of February to 4th week of September and higher population was during 46th to 51st standard week, during 3rd week of November to 3rd week of December and peak population (6.06/3 leaves) was recorded on 46th standard week that is on 3rd week of November. Correlation studies revealed that mite population had a significant negative correlation with temperature and non-significant positive correlation with relative humidity. This indicates that activity of mites population increase with the rise of relative humidity and decrease with the rise of temperature. Tobacco leaf extracts was found most effective against mite providing 40.51% suppression, closely followed by extracts of Spilanthes (39.06% suppression). Extracts of Garlic and extracts of Polygonum plant gave moderate results, recording about 38.10% and 37.78% mite suppression respectively. The polygonum (Polygonum hydropiper) plant (floral parts), pongamia (Pongamia pinnata) leaves, garlic (Allium sativum), spilanthes (Spilanthes paniculata) (floral parts) were extracted in methanol. Synthetic insecticides contaminate plant leaves with the toxic chemicals. Plant extracts are of biological origin having low or no hazardous effect on health and environment and so can be incorporated in organic cultivation.

Keywords: Abiotic factors, incidence, botanical extracts, organic cultivation, silk industry

Procedia PDF Downloads 127

Authors: Thomas Chinedu Aniokete, Gordian Onyebuchukwu Mbah, Michael Daramola

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A sustainable NaOH integrated hydrothermal protocol was developed for the synthesis of waste-derived hydroxy sodalite catalysts for transesterification of waste animal fat (WAF) with a high per cent free fatty acid (FFA) to biodiesel. In this work, hydroxy sodalite catalyst was synthesized from two complex waste materials namely coal fly ash (CFA) and waste industrial brine (WIB). Measured amounts of South African CFA and WIB obtained from a coal mine field were mixed with NaOH solution at different concentrations contained in secured glass vessels equipped with magnetic stirrers and formed consistent slurries after aging condition at 47 oC for 48 h. The slurries were then subjected to hydrothermal treatments at 140 oC for 48 h, washed thoroughly and separated by the action of a centrifuge on the mixture. The resulting catalysts were calcined in a muffle furnace for 2 h at 200 oC and subsequently characterized for different effects using X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR), and Bennett Emmet Teller (BET) adsorption-desorption techniques. The produced animal fat methyl ester (AFME) was analyzed using the gas chromatography-mass spectrometry (GC-MS) method. Results of the investigation indicate profoundly an enhanced catalyst purity, textural property and desired morphology due to the action of NaOH. Similarly, the performance evaluation with respect to catalyst activity reveals a high catalytic conversion efficiency of 98 % of the high FFA WAF to biodiesel under the following reaction conditions; a methanol-to-WAF ratio of 15:1, amount of SOD catalyst of 3 wt % with a stirring speed of 300-500 rpm, a reaction temperature of 60 oC and a reaction time of 8 h. There was a recovered 96 % stable catalyst after reactions and potentially recyclable, thus contributing to the economic savings to the process that had been a major bottleneck to the production of biodiesel. This NaOH route for synthesizing waste-derived hydroxy sodalite (SOD) catalyst is a sustainable and eco-friendly technology that speaks directly to the global quest for renewable-fossil fuel controversy enforcing sustainable development goal 7.

Keywords: coal fly ash, waste industrial brine, waste-derived hydroxy sodalite catalyst, sodium hydroxide, biodiesel, transesterification, biomass conversion

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Authors: Josline Salib, Sayed El-Toumy, Abeer Salama, Enayat Omara, Emad Hassan

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Diabetic nephropathy is an important cause of morbidity and mortality and is now among the most common causes of end-stage renal failure (ESRF) in developed countries. Thus, the aim of the present study was to investigate the phenolic compounds content of Cichorium intybus aerial parts extracts as well as the therapeutic effects on diabetic nephropathy, oxidative stress, and anti-inflammatory by characterizing biochemical, histopathological changes and immunohistochemistry in an experimental diabetic rat model as compared with Amaryl. Ten known compounds of flavonoids, coumarins and phenolic acid derivatives were isolated from the C. intybus aqueous methanolic extract. Structures of the isolated compounds were established by chromatography, UV and 1D⁄2D 1H⁄ 13C spectroscopy. The aqueous methanol extract of C. intybus aerial parts was administered to Streptozotocin diabetes rats at doses (100 and 200 mg/kg) for 21 days. After treatment, blood glucose, serum insulin, urea, creatinine, and TNF-α were evaluated. Enzymatic scavengers including catalase (CAT), glutathione (GSH), malondialdehyde (MDA) and nitric oxide (NO) were determined to evaluate the oxidative status in the renal tissue. Diabetic rats treated with C. intybus extract showed a dose-dependent reduction of fasting blood glucose and kidney antioxidant status in comparison to the diabetic control group. The extract was able to enhance the antioxidant defenses of the kidney by increasing the reduced GSH and CAT content and decreasing MDA content in addition to significantly decreasing kidney nitric oxide content compared to diabetic control rats. Furthermore, the histopathological findings in C. intybus extract administered rats were observed at markedly lesser extent than the diabetic control group. Also, inducible nitric oxide synthase (iNOS) levels were decreased significantly after the administration of high-dose C. intybus extract in diabetic rats. Showing significant antihyperglycemic and antioxidant properties of C. intybus aerial parts extract, which is attributed to its polyphenolic content, may offer a potential source for the treatment of diabetes.

Keywords: antioxidant activity, anti-diabetic nephropathy, cichorium intybus aerial parts, phenolic compounds

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Authors: Aashutosh Garg, Ankur Rajpal, A. A. Kazmi

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The efficiency of an on-site wastewater treatment system named Johkasou was evaluated based on its pollutant removal efficiency over 10 months. This system was installed at IIT Roorkee and had a capacity of treating 7 m3/d of sewage water, sufficient for a group of 30-50 people. This system was fed with actual wastewater through an equalization tank to eliminate the fluctuations throughout the day. Methanol and ammonium chloride was added into this equalization tank to increase the Chemical Oxygen Demand (COD) and ammonia content of the influent. The outlet from Johkasou is sent to a tertiary unit consisting of a Pressure Sand Filter and an Activated Carbon Filter for further treatment. Samples were collected on alternate days from Monday to Friday and the following parameters were evaluated: Chemical Oxygen Demand (COD), Biochemical Oxygen Demand (BOD), Total Suspended Solids (TSS), and Total Nitrogen (TN). The Average removal efficiency for Chemical Oxygen Demand (COD), Biochemical Oxygen Demand (BOD), Total Suspended Solids (TSS), and Total Nitrogen (TN) was observed as 89.6, 97.7, 96, and 80% respectively. The cost of treating the wastewater comes out to be Rs 23/m3 which includes electricity, cleaning and maintenance, chemical, and desludging costs. Tests for the coliforms were also performed and it was observed that the removal efficiency for total and fecal coliforms was 100%. The sludge generation rate is approximately 20% of the BOD removal and it needed to be removed twice a year. It also showed a very good response against the hydraulic shock load. We performed vacation stress analysis on the system to evaluate the performance of the system when there is no influent for 8 consecutive days. From the result of stress analysis, we concluded that system needs a recovery time of about 48 hours to stabilize. After about 2 days, the system returns again to original conditions and all the parameters in the effluent become within the limits of National Green Tribunal (NGT) standards. We also performed another stress analysis to save the electricity in which we turned the main aeration blower off for 2 to 12 hrs a day and the results showed that we can turn the blower off for about 4-6 hrs a day and this will help in reducing the electricity costs by about 25%. It was concluded that the Johkasou system can remove a sufficient amount of all the physiochemical parameters tested to satisfy the prescribed limit set as per Indian Standard.

Keywords: on-site treatment, domestic wastewater, Johkasou, nutrient removal, pathogens removal

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Authors: Siti Aisya Gany, Swee Ching Tan, Sook Yee Gan

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Diminished antioxidant defense or increased production of reactive oxygen species in the biological system can result in oxidative stress which may lead to various neurodegenerative diseases including Alzheimer’s disease (AD). Microglial activation also contributes to the progression of AD by producing several pro-inflammatory cytokines, nitric oxide (NO), and prostaglandin E2 (PGE2). Oxidative stress and inflammation have been reported to be possible pathophysiological mechanisms underlying AD. In addition, the cholinergic hypothesis postulates that memory impairment in patient with AD is also associated with the deficit of cholinergic function in the brain. Although a number of drugs have been approved for the treatment of AD, most of these synthetic drugs have diverse side effects and yield relatively modest benefits. Marine algae have great potential in pharmaceutical and biomedical applications as they are valuable sources of bioactive properties such as anti-coagulation, anti-microbial, anti-oxidative, anti-cancer and anti-inflammatory. Hence, this study aimed to provide an overview of the properties of Malaysian seaweeds (Padina australis, Sargassum polycystum and Caulerpa racemosa) in inhibiting oxidative stress, neuroinflammation and cholinesterase enzymes. All tested samples significantly exhibit potent DPPH and moderate Superoxide anion radical scavenging ability (P<0.05). Hexane and methanol extracts of S. polycystum exhibited the most potent radical scavenging ability with IC50 values of 0.1572 ± 0.004 mg/ml and 0.8493 ± 0.02 for DPPH and ABTS assays, respectively. Hexane extract of C. racemosa gave the strongest superoxide radical inhibitory effect (IC50 of 0.3862± 0.01 mg/ml). Most seaweed extracts significantly inhibited the production of cytokine (IL-6, IL-1 β, TNFα) and NO in a concentration-dependent manner without causing significant cytotoxicity to the lipopolysaccharide (LPS)-stimulated microglia cells (P<0.05). All extracts suppressed cytokine and NO level by more than 80% at the concentration of 0.4mg/ml. In addition, C. racemosa and S. polycystum also showed anti-acetylcholinesterase activities with the IC50 values ranging from 0.086-0.115 mg/ml. Moreover, C. racemosa and P. australis were also found to be active against butyrylcholinesterase with IC50 values ranging from 0.118-0.287 mg/ml.

Keywords: anti-cholinesterase, anti-oxidative, neuroinflammation, seaweeds

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Authors: Omoruyi G. Idemudia, Alexander P. Sadimenko

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The need for the development of new sustainable bioactive compounds with unique properties that can become potential replacement for commonly used medicinal drugs has continued to gain tremendous research concerns because of the problems of disease resistant to these medicinal drugs and their toxicity effects. NOS-donor heterocycles are particularly of interest as they have showed good pharmacological activities in the midst of their interesting chelating properties towards metal ions, an important characteristic for transition metal based drugs design. These new compounds have also gained application as dye sensitizers in solar cell panels for the generation of renewable solar energy, as greener water purification polymer for supply and management of clean water and as catalysts which are used to reduce the amount of pollutants from industrial reaction processes amongst others, because of their versatile properties. Di-ketone acylpyrazolones and their azomethine schiff bases have been employed as pharmaceuticals as well as analytical reagents, and their application as transition metal complexes have being well established. In this research work, a new 4-propyl-3-methyl-1-phenyl-2-pyrazolin-5-one-thiosemicarbazone was synthesized from the reaction of 4-propyl-3-methyl-1-phenyl-2-pyrazolin-5-one and thiosemicarbazide in methanol. The pure isolate of the thiosemicarbazone was further reacted with aqueous solutions of palladium and platinum salts to obtain their metal complexes, in an effort towards the discovery of transition metal based synthetic drugs. These compounds were characterized by means of analytical, spectroscopic, thermogravimetric analysis TGA, as well as x-ray crystallography. 4-propyl-3-methyl-1-phenyl-2-pyrazolin-5-one thiosemicarbazone crystallizes in a triclinic crystal system with a P-1 (No. 2) space group according to x-ray crystallography. The tridentate NOS ligand formed a tetrahedral geometry on coordinating with metal ions. Reported compounds showed varying antioxidant free radical scavenging activities against 2, 2-diphenyl-1-picrylhydrazyl DPPH radical at 100, 200, 300, 400 and 500 µg/ml concentrations. The platinum complex have shown a very good antioxidant property against DPPH with an IC50 of 76.03 µg/ml compared with standard ascorbic acid (IC50 of 74.66 µg/ml) and as such have been identified as a potential anticancer candidate.

Keywords: acylpyrazolone, free radical scavenging activities, tridentate ligand, x-ray crystallography

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Authors: Magdalena Smoktunowicz, Renata Wawrzyniak, Malgorzata Waleron, Krzysztof Waleron

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Pectobacterium spp. were previously classified into the Erwinia genus founded in 1917 to unite at that time all Gram-negative, fermentative, nonsporulating and peritrichous flagellated plant pathogenic bacteria. After work of Waldee (1945), on Approved Lists of Bacterial Names and bacteriology manuals in 1980, they were described either under the species named Erwinia or Pectobacterium. The Pectobacterium genus was formally described in 1998 of 265 Pectobacterium strains. Currently, there are 21 species of Pectobacterium bacteria, including Pectobacterium betavasculorum since 2003, which caused soft rot on sugar beet tubers. Based on the biochemical experiments carried out for this, it is known that these bacteria are gram-negative, catalase-positive, oxidase-negative, facultatively anaerobic, using gelatin and causing symptoms of soft rot on potato and sugar beet tubers. The mere fact of growing on sugar beet may indicate a metabolism characteristic only for this species. Metabolomics, broadly defined as the biology of the metabolic systems, which allows to make comprehensive measurements of metabolites. Metabolomics, in combination with genomics, are complementary tools for the identification of metabolites and their reactions, and thus for the reconstruction of metabolic networks. The aim of this study was to apply the GC-MS-based untargeted metabolomics to study the metabolism of P. betavasculorum in different growing conditions. The metabolomic profiles of biomass and biomass media were determined. For sample preparation the following protocol was used: extraction with 900 µl of methanol: chloroform: water mixture (10: 3: 1, v: v) were added to 900 µl of biomass from the bottom of the tube and up to 900 µl of nutrient medium from the bacterial biomass. After centrifugation (13,000 x g, 15 min, 4oC), 300µL of the obtained supernatants were concentrated by rotary vacuum and evaporated to dryness. Afterwards, two-step derivatization procedure was performed before GC-MS analyses. The obtained results were subjected to statistical calculations with the use of both uni- and multivariate tests. The obtained results were evaluated using KEGG database, to asses which metabolic pathways are activated and which genes are responsible for it, during the metabolism of given substrates contained in the growing environment. The observed metabolic changes, combined with biochemical and physiological tests, may enable pathway discovery, regulatory inference and understanding of the homeostatic abilities of P. betavasculorum.

Keywords: GC-MS chromatograpfy, metabolomics, metabolism, pectobacterium strains, pectobacterium betavasculorum

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Authors: Débora P. Jaeschke, Eduardo A. Merlo, Rosane Rech, Giovana D. Mercali, Ligia D. F. Marczak

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Carotenoids are high value added pigments that can be alternatively extracted from some microalgae species. However, the application of carotenoids synthetized by microalgae is still limited due to the utilization of organic toxic solvents. In this context, studies involving alternative extraction methods have been conducted with more sustainable solvents to replace and reduce the solvent volume and the extraction time. The aim of the present work was to evaluate the extraction time of carotenoids from the microalgae Heterochlorella luteoviridis using moderate electric field (MEF) as a pre-treatment to the extraction. The extraction methodology consisted of a pre-treatment in the presence of MEF (180 V) and ethanol (25 %, v/v) for 10 min, followed by a diffusive step performed for 50 min using a higher ethanol concentration (75 %, v/v). The extraction experiments were conducted at 30 °C and, to keep the temperature at this value, it was used an extraction cell with a water jacket that was connected to a water bath. Also, to enable the evaluation of MEF effect on the extraction, control experiments were performed using the same cell and conditions without voltage application. During the extraction experiments, samples were withdrawn at 1, 5 and 10 min of the pre-treatment and at 1, 5, 30, 40 and 50 min of the diffusive step. Samples were, then, centrifuged and carotenoids analyses were performed in the supernatant. Furthermore, an exhaustive extraction with ethyl acetate and methanol was performed, and the carotenoids content found for this analyses was considered as the total carotenoids content of the microalgae. The results showed that the application of MEF as a pre-treatment to the extraction influenced the extraction yield and the extraction time during the diffusive step; after the MEF pre-treatment and 50 min of the diffusive step, it was possible to extract up to 60 % of the total carotenoids content. Also, results found for carotenoids concentration of the extracts withdrawn at 5 and 30 min of the diffusive step did not presented statistical difference, meaning that carotenoids diffusion occurs mainly in the very beginning of the extraction. On the other hand, the results for control experiments showed that carotenoids diffusion occurs mostly during 30 min of the diffusive step, which evidenced MEF effect on the extraction time. Moreover, carotenoids concentration on samples withdrawn during the pre-treatment (1, 5 and 10 min) were below the quantification limit of the analyses, indicating that the extraction occurred in the diffusive step, when ethanol (75 %, v/v) was added to the medium. It is possible that MEF promoted cell membrane permeabilization and, when ethanol (75 %) was added, carotenoids interacted with the solvent and the diffusion occurred easily. Based on the results, it is possible to infer that MEF promoted the decrease of carotenoids extraction time due to the increasing of the permeability of the cell membrane which facilitates the diffusion from the cell to the medium.

Keywords: moderate electric field (MEF), pigments, microalgae, ethanol

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Authors: Aleksandr Kalinenko, Igor Vysotskiy, Sergey Malopheyev, Sergey Mironov, Rustam Kaibyshev

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From a thermo-mechanical standpoint, friction-stir welding (FSW) represents a unique combination of very large strains, high temperature and relatively high strain rate. The material behavior under such extreme deformation conditions is not studied well and thus, the microstructural examinations of the friction-stir welded materials represent an essential academic interest. Moreover, a clear understanding of the microstructural mechanisms operating during FSW should improve our understanding of the microstructure-properties relationship in the FSWed materials and thus enables us to optimize their service characteristics. Despite extensive research in this field, the microstructural behavior of some important structural materials remains not completely clear. In order to contribute to this important work, the present study was undertaken to examine the grain structure evolution during the FSW of 6061-T6 aluminum alloy. To provide an in-depth insight into this process, the electron backscatter diffraction (EBSD) technique was employed for this purpose. Microstructural observations were conducted by using an FEI Quanta 450 Nova field-emission-gun scanning electron microscope equipped with TSL OIMTM software. A suitable surface finish for EBSD was obtained by electro-polishing in a solution of 25% nitric acid in methanol. A 15° criterion was employed to differentiate low-angle boundaries (LABs) from high-angle boundaries (HABs). In the entire range of the studied FSW regimes, the grain structure evolved in the stir zone was found to be dominated by nearly-equiaxed grains with a relatively high fraction of low-angle boundaries and the moderate-strength B/-B {112}<110> simple-shear texture. In all cases, the grain-structure development was found to be dictated by an extensive formation of deformation-induced boundaries, their gradual transformation to the high-angle grain boundaries. Accordingly, the grain subdivision was concluded to the key microstructural mechanism. Remarkably, a gradual suppression of this mechanism has been observed at relatively high welding temperatures. This surprising result has been attributed to the reduction of dislocation density due to the annihilation phenomena.

Keywords: electron backscatter diffraction, friction-stir welding, heat-treatable aluminum alloys, microstructure

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Authors: Esmeralda Chávez Vargas, M. de la L. Olvera, A. Maldonado

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The doping it is believed as follows, at high concentration fluorine in ZnO: F films is incorporated to the lattice by substitution of O-2 ions by F-1 ions; at middle fluorine concentrations, F ions may form interstitials, whereas for low concentrations it is increased the carriers and mobility could be explained by the surface passivation effect of fluorine. ZnO:F thin films were deposited on sodocalcic glass substratesat 425 °C , 450°C, 475 during 8, 12, 15 min from a 0.2 M solution. Doping concentration in the starting solutions was varied, namely, [F]/[F+Zn] = 0, 5, 15, 30, 45, 60, and 90 at. %; solvent composition was varied as well, 100:100; 50:50; 100:50(acetic acid: water: methanol ratios, in volume). In this work it is reported the characterization results of fluorine doped zinc oxide (ZnO:F) thin films deposited by the ultrasonic spray pyrolysis technique, using zinc acetate and ammonium fluorine as Zn an F precursors, respectively. The effect of varying the fluorine concentration in the starting solutions, the solvent composition, and the ageing time of the starting solutions, on the electrical resistivity, optical transmittance, structure and surface morphology was analyzed. In order to have a quantitative evaluation of the ZnO:F thin films for its application as transparent electrodes, the Figure of Merit was estimated from the Haacke´s formula. After a thoroughly study, it can be found that optimal conditions for the deposition of transparent and conductive ZnO:F thin films on sodocalcic substrates, were as follows; substrate temperature: solution molar concentration 0.2, doping concentration in the starting solution of [F]/[Zn]= 60 at. %, (water content)/(acetic acid) in starting solution: [H2O/ CH3OH]= 50:50, substrate temperature: 450 °C. The effects of aging of the starting solution has also been analyzed thoroughly and it has been found a dramatic effect on the electric resistivity of the material, aged by 40 days, show an electrical resitivity as low as 120 Ω/□, with a transmittance around 80% in the visible range. X-ray diffraction spectra show a polycrystalline of ZnO (wurtzite structure) where the amount of fluorine doping affects to preferential orientation (002 plane). Therefore, F introduction in lattice is by the substitution of O-2 ions by F-1 ions. The results show that ZnO:F thin films are potentially adequate for application as transparent conductive oxide in thin film solar cells.

Keywords: TCOs, transparent electrodes, ultrasonic spray pyrolysis, zinc oxide, ZnO:F

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Authors: H. Minor-Pérez, A. M. Mota-Silva, S. Ortiz-Barrios

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Substances of vegetable origin with antioxidant capacity have a high potential for application on the conservation of some foods, can prevent or reduce for example oxidation of lipids. However a food is a complex system whose wide variety of components wich can reduce or eliminate this antioxidant capacity. The antioxidant activity can be determined with the ABTS technique. The radical ABTS+ is generated from the acid 2, 2´ - Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). This radical is a composite color bluish-green, stable and with a spectrum of absorption into the UV-visible. The addition of antioxidants causes discoloration, value that can be reported as a percentage of inhibition of the cation radical ABTS+. The objective of this study was evaluated the effect of the combination of the pH and the essential oil of cinnamon (EOC) on inhibition of the radical ABTS+, using statistical design of central point (Design Expert) to obtain mathematical models that describe this phenomenon. Were evaluated 17 treatments with combinations of pH 5, 6 and 7 (citrate-phosphate buffer) and the concentration of essential oil of cinnamon (C): 0 µg/mL, 100 µg/mL and 200 µg/mL. The samples were analyzed using the ABTS technique. The reagent was dissolved in methanol 80% to standardized the absorbance to 0.7 +/- 0.1 at 754 nm. Then samples were mixed with reagent standardized ABTS and after 1 min and 7 min absorbance was read for each treatment at 754 nm. Was used a curve pattern with vitamin C and reported the values as inhibition (%) of radical ABTS+. The statistical analysis shows the experimental results were adjusted to a quadratic model, to the times of 1 min and 7 min. This model describes the influence of the factors investigated independently: pH and cinnamon essential oil (µg/mL) and the effect of the interaction between pH*C, as well as the square of the pH2 and C2. The model obtained was Y = 10.33684 - 3.98118*pH + 1.17031*C + 0.62745*pH2 - 3.26675*10-3*C2 - 0.013112*pH*C, where Y is the response variable. The coefficient of determination was 0.9949 for 1 min. The equation was obtained at 7 min and = - 10.89710 + 1.52341*pH + 1.32892*C + 0.47953*pH2 - 3.56605*10- *C2 - 0.034687*pH*C. The coefficient of determination was 0.9970. This means that only 1% of the total variation is not explained by the developed models. At 100 µg/mL of EOC was obtained an inhibition percentage of 80%, 84% and 97% for the pH values of 5,6 and 7 respectively, while a value of 200 µg/mL the inhibition (%) was very similar for the treatments. In these values of pH was obtained an inhibition close 97%. In conclusion the pH does not have a significant effect on the antioxidant capacity, while the concentration of EOC was decisive for the antioxidant capacity. The authors acknowledge the funding provided by the CONACYT for the project 131998.

Keywords: antioxidant activity, ABTS technique, essential oil of cinnamon, mathematical models

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Authors: Carlos Y. Soto, Camila A. Lota, G. Astrid Garzón

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Bacterial biofilms cause an ongoing problem for food safety. They are formed when microorganisms aggregate to form a community that attaches to solid surfaces. Biofilms increase the resistance of pathogens to cleaning, disinfection and antibacterial products. This resistance gives rise to problems for human health, industry, and agriculture. At present, plant extracts rich in polyphenolics are being investigated as natural alternatives to degrade bacterial biofilms. The pomace of the tropical Berry Vaccinium meridionale S. contains high amounts of phenolic compounds. Therefore, in the current study, the antimicrobial and antibiofilm effects of extracts from the pomace of Vaccinium meridionale S. were tested on three foodborne pathogens: Enterohaemorrhagic Escherichia coli O157:H7 (ATCC®700728TM), Staphylococcus aureus subsp. aureus (ATCC® 6538TM), and Salmonella enterica serovar Enteritidis (ATCC® 13076TM). Microwave-assisted extraction was used to extract polyphenols with aqueous methanol (80% v/v) at a solid to solvent ratio of 1:10 (w/v) for 20 min. The magnetic stirring was set at 400 rpm, and the microwave power was adjusted to 400 W. The antimicrobial effect of the extract was assessed by determining the half maximal inhibitory concentration (IC50) against the three food poisoning pathogens at concentrations ranging from 50 to 2,850 μg gallic acid equivalents (GAE)/mL of the extract. Biofilm inhibition was assessed using a crystal violet assay applying the same range of concentration. Three replications of the experiments were carried out, and all analyses were run in triplicate. IC50 values were determined using the GraphPad Prism8® program. Significant differences (P<0.05) among means were identified using one-factor analysis of variance (ANOVA) and the post-hoc least significant difference (LSD) test using the Statgraphics plus program, version 2.1.There was significant difference among the mean IC50 values for the tested bacteria. The IC50 for S. aureus was 48 ± 9 μg GAE/mL, followed by 123 ± 49 μg GAE/mL for Salmonella and 376 ± 32 μg GAE/mL for E. coli. The percent inhibition of the extract on biofilm formation was significantly higher for S. aureus (85.8  0.3), followed by E. coli (74.5  1.0) and Salmonella (53.6  9.7). These findings suggest that polyphenolic extracts obtained from the pomace of V. meridionale S. might be used as natural antimicrobial and anti-biofilm natural agents, effective against S. aureus, E. coli and Salmonella enterica.

Keywords: antibiofilm, antimicrobial, E. coli, S. aureus, salmonella, IC50, pomace, V. meridionale

Procedia PDF Downloads 51

Authors: Vipin Kumar Verma, Neeta Sehgal, Om Prakash

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Cyprinus carpio has a wide spread occurrence in the lakes and rivers of Europe and Asia. Heavy losses in natural environment due to anthropogenic activities, including pollution as well as pathogenic diseases have landed this fish in IUCN red list of vulnerable species. The significance of a suitable diet in preserving the health status of fish is widely recognized. In present study, artificial feed supplemented with leaves of two weed plants, Eichhornia crassipes and Ricinus communis were evaluated for their role on the fish immune system. To achieve this objective fish were acclimatized to laboratory conditions (25 ± 1 °C; 12 L: 12D) for 10 days prior to start of experiment and divided into 4 groups: non-challenged (negative control= A), challenged [positive control (B) and experimental (C & D)]. Group A, B were fed with non-supplemented feed while group C & D were fed with feed supplemented with 5% Eichhornia crassipes and 5% Ricinus communis respectively. Supplemented feeds were evaluated for their effect on growth, health, immune system and disease resistance in fish when challenged with Vibrio harveyi. Fingerlings of C. carpio (weight, 2.0±0.5 g) were exposed with fresh overnight culture of V. harveyi through bath immunization (concentration 2 Χ 105) for 2 hours on 10 days interval for 40 days. The growth was monitored through increase in their relative weight. The rate of mortality due to bacterial infection as well as due to effect of feed was recorded accordingly. Immune response of fish was analyzed through differential leucocyte count, percentage phagocytosis and phagocytic index. The effect of V. harveyi on fish organs were examined through histo-pathological examination of internal organs like spleen, liver and kidney. The change in the immune response was also observed through gene expression analysis. The antioxidant potential of plant extracts was measured through DPPH and FRAP assay and amount of total phenols and flavonoids were calculates through biochemical analysis. The chemical composition of plant’s methanol extracts was determined by GC-MS analysis, which showed presence of various secondary metabolites and other compounds. Investigation revealed immuno-modulatory effect of plants, when supplemented with the artificial feed of fish.

Keywords: immuno-modulation, gc-ms, Cyprinus carpio, Eichhornia crassipes, Ricinus communis

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Authors: Jung-En Kuan, Whei-Fen Wu

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In this study, a symbiotic bacteria from yellow mealworm's (Tenebrio molitor) mid-gut was isolated with characteristics of growth on minimal-tributyrin medium. After a PCR-amplification of its 16s rDNA, the resultant nucleotide sequences were then analyzed by schemes of the phylogeny trees. Accordingly, it was designated as Pseudomonas nitroreducens D-01. Next, by searching the lipolytic enzymes in its protein data bank, one of those potential lipolytic α/β hydrolases was identified, again using PCR-amplification and nucleotide-sequencing methods. To construct an expression of this lipolytic gene in plasmids, the target-gene primers were then designed, carrying the C-terminal his-tag sequences. Using the vector pET21a, a recombinant lipolytic hydrolase D gene with his-tag nucleotides was successfully cloned into it, of which the lipolytic D gene is under a control of the T7 promoter. After transformation of the resultant plasmids into Eescherichia coli BL21 (DE3), an IPTG inducer was used for the induction of the recombinant proteins. The protein products were then purified by metal-ion affinity column, and the purified proteins were found capable of forming a clear zone on tributyrin agar plate. Shortly, its enzyme activities were determined by degradation of p-nitrophenyl ester(s), and the substantial yellow end-product, p-nitrophenol, was measured at O.D.405 nm. Specifically, this lipolytic enzyme efficiently targets p-nitrophenyl butyrate. As well, it shows the most reactive activities at 40°C, pH 8 in potassium phosphate buffer. In thermal stability assays, the activities of this enzyme dramatically drop when the temperature is above 50°C. In metal ion assays, MgCl₂ and NH₄Cl induce the enzyme activities while MnSO₄, NiSO₄, CaCl₂, ZnSO₄, CoCl₂, CuSO₄, FeSO₄, and FeCl₃ reduce its activities. Besides, NaCl has no effects on its enzyme activities. Most organic solvents decrease the activities of this enzyme, such as hexane, methanol, ethanol, acetone, isopropanol, chloroform, and ethyl acetate. However, its enzyme activities increase when DMSO exists. All the surfactants like Triton X-100, Tween 80, Tween 20, and Brij35 decrease its lipolytic activities. Using Lineweaver-Burk double reciprocal methods, the function of the enzyme kinetics were determined such as Km = 0.488 (mM), Vmax = 0.0644 (mM/min), and kcat = 3.01x10³ (s⁻¹), as well the total efficiency of kcat/Km is 6.17 x10³ (mM⁻¹/s⁻¹). Afterwards, based on the phylogenetic analyses, this lipolytic protein is classified to type IV lipase by its homologous conserved region in this lipase family.

Keywords: enzyme, esterase, lipotic hydrolase, type IV

Procedia PDF Downloads 125

Authors: Bunindro Nameirakpam, Sonia Sougrapakam, Shannon B. Olsson, Rajashekar Yallappa

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Natural sources for new pesticidal compounds hold promise in view of their eco-friendly nature, selectivity and mammalian safety. Despite a large number of plants that show insecticidal activity and diversity of natural chemistry with inherent eco-friendly nature, newer classes of insecticides have eluded discovery. Artemisia vulgaris, known as Mugwort, is a universal herb used for folk medicine and religious purposes throughout the ancient world. In India, the essential oils of Artemisia vulgaris are used for its insecticidal, anti parasiticidal and antimicrobial properties. Traditionally, the dried leaves of Artemisia vulgaris are used to repel insects as well as rats in and around the granaries in the North-East India. Artemisia vulgaris collected during November from different ecological sites were studied for the bio-pesticidal utility against the stored grain pests. The insecticidal activities were found in the crude extracts of n-hexane and methanol from the samples collected in Sikkim and Manipur respectively. Using silica gel column chromatography protocol, we have isolated one novel bioactive molecule from the aerial parts of Artemisia vulgaris L based on various physical-chemical and spectroscopic techniques (IR, 1H NMR, 13C NMR and mass). The novel bioactive molecule is highly toxic and very low concentration (4.35 µg/l) is needed to control the stored product insects. In additional experiment results clearly showed the involvement of sodium pumps inhibition in the insecticidal action of purified compound in the Sitophilus oryzae. The knockdown activity of the purified compound is concomitant with the in vivo inhibition of Na+/ K+- ATPase. Further, our study showed insignificant differences in the seed germination of control and the treated grains. The lack of adverse effect of the novel bioactive molecule on the seed germination is highly desirable for seed/grain protectant and showing the potential to be developed as possible natural fumigants for the control of stored grain pests. The novel bioactive molecule is selective insecticide with a high margin of safety to mammals and showed promise as novel biopesticide candidate for grain protection. It is believed that Bio-pesticides can serve as the most important pest management tools as far as global safety is concerned.

Keywords: Indian traditional plant, Artemisia vulgaris, bio-pesticides, Na+/ K+- ATPase, seed germination

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Authors: Soo Im Chung, Lara Marie Pangan Lo, Yao Cheng Zhang, Su Jin Nam, Xingyue Jin, Mi Young Kang

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Rice (Oryza sativa L.) is one of the most widely consumed grains. Due to the growing number of demand as a potential functional food and nutraceutical source and the increasing awareness of people towards healthy diet and good quality of living, more researches dwell upon the development of new rice cultivars for population consumption. However, studies on the antioxidant capacities of newly developed rice were limited as well as the effects of germination in these rice cultivars. Therefore, this study aimed to focus on analysis of the antioxidant potential of pre-germinated and germinated pigmented rice cultivars in South Korea such as purple cultivar Superjami (SJ) and red cultivar Super hongmi (SH) in comparison with the non-pigmented Normal Brown (NB) Rice. The powdered rice grain samples were extracted with 80% methanol and their antioxidant activities were determined. The Results showed that pre-germinated pigmented rice cultivars have higher Fe2+ Chelating Ability (Fe2+), Reducing Power (RP), 2,2´-azinobis[3-ethylbenzthiazoline]-6-sulfonic acid (ABTS) radical scavenging and Superoxide Dismutase activity than the control NB rice. Moreover, it is revealed that germination process induced a significant increased in the antioxidant activities of all the rice samples regardless of their strains. Purple rice SJ showed greater Fe2+ (88.82 + 0.53%), RP (0.82 + 0.01) , ABTS (143.63 + 2.38 mg VCEAC/100 g) and SOD (59.31 + 0.48%) activities than the red grain SH and the control NB having the lowest antioxidant potential among the three (3) rice samples examined. The Effective concentration at 50% (EC50) of 1, 1-Diphenyl-2-picrylhydrazyl (DPPH) and Hydroxyradical (-OH) Scavenging activity for the rice samples were also obtained. SJ showed lower EC50 in terms of its DPPH (3.81 + 0.15 mg/mL) and –OH (5.19 + 0.08 mg/mL) radical scavenging activities than the red grain SH and control NB rice indicating that at lower concentrations, it can readily exhibit antioxidant effects against reactive oxygen species (ROS). These results clearly suggest the higher antioxidant potential of pigmented rice varieties as compared with the widely consumed NB rice. Also, it is revealed in the study that even at lower concentrations, pigmented rice varieties can exhibit their antioxidant activities. Germination process further enhanced the antioxidant capacities of the rice samples regardless of their types. With these results at hand, these new rice varieties can be further developed as a good source of bio functional elements that can help alleviate the growing number of cases of metabolic disorders.

Keywords: antioxidant capacity, germinated rice, pigmented rice, super hongmi, superjami

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Authors: Julia R. Beulig, Stefan Ohla, Detlev Belder

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In contrast to off-line analytical methods, lab-on-a-chip technology delivers direct information about the observed reaction. Therefore, microfluidic devices make an important scientific contribution, e.g. in the field of synthetic chemistry. Herein, the rapid generation of analytical data can be applied for the optimization of chemical reactions. These microfluidic devices enable a fast change of reaction conditions as well as a resource saving method of operation. In the presented work, we focus on the investigation of multiphase regimes, more specifically on a biphasic microfluidic droplet systems. Here, every single droplet is a reaction container with customized conditions. The biggest challenge is the rapid qualitative and quantitative readout of information as most detection techniques for droplet systems are non-specific, time-consuming or too slow. An exception is the electrospray mass spectrometry (ESI-MS). The combination of a reaction screening platform with a rapid and specific detection method is an important step in droplet-based microfluidics. In this work, we present a novel approach for synthesis optimization on the nanoliter scale with direct ESI-MS detection. The development of a droplet-based microfluidic device, which enables the modification of different parameters while simultaneously monitoring the effect on the reaction within a single run, is shown. By common soft- and photolithographic techniques a polydimethylsiloxane (PDMS) microfluidic chip with different functionalities is developed. As an interface for the MS detection, we use a steel capillary for ESI and improve the spray stability with a Teflon siphon tubing, which is inserted underneath the steel capillary. By optimizing the flow rates, it is possible to screen parameters of various reactions, this is exemplarity shown by a Domino Knoevenagel Hetero-Diels-Alder reaction. Different starting materials, catalyst concentrations and solvent compositions are investigated. Due to the high repetition rate of the droplet production, each set of reaction condition is examined hundreds of times. As a result, of the investigation, we receive possible reagents, the ideal water-methanol ratio of the solvent and the most effective catalyst concentration. The developed system can help to determine important information about the optimal parameters of a reaction within a short time. With this novel tool, we make an important step on the field of combining droplet-based microfluidics with organic reaction screening.

Keywords: droplet, mass spectrometry, microfluidics, organic reaction, screening

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Authors: Leonardo Parra, Daniel Martínez, Jorge Pizarro, Fernando Ortega, Manuel Chacón-Fuentes, Andrés Quiroz

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Control of Dalaca pallens or blackworms, one of the most important hypogeous pest in grassland in southern Chile, is based on the use of broad-spectrum insecticides such as organophosphates and pyrethroids. However, the rapid development of insecticide resistance in field populations of this insect and public concern over the environmental impact of these insecticides has resulted in the search for other control methods. Specifically, the use of endophyte fungi for controlling pest has emerged as an interesting and promising strategy. Endophytes from ryegrass (Lolium perenne), establish a biotrophic relationship with the host, defined as mutualistic symbiosis. The plant-fungi association produces alkaloids where peramine is the main toxic substance against Listronotus bonariensis, the most important epigean pest of ryegrass. Nevertheless, the effect of peramina on others pest insects, such as D. pallens, to our knowledge has not been studied, and also its possible metabolization in the body of the larvae. Therefore, we addressed the following research question: Do larvae of D. pallens store peramine after consumption of ryegrass endophyte infected (E+)? For this, specimens of blackworms were fed with ryegrass plant of seven experimental lines and one commercial cultivar endophyte free (E-) sown at the Instituto de Investigaciones Agropecuarias Carillanca (Vilcún, Chile). Once the feeding period was over, ten larvae of each treatment were examined. Individuals were dissected, and their gut was removed to exclude any influence of remaining material. The rest of the larva's body was dried at 60°C by 24-48 h and ground into a fine powder using a mortar. 25 mg of dry powder was transferred to a microcentrifuge tube and extracted in 1 mL of a mixture of methanol:water:formic acid. Then, the samples were centrifuged at 16,000 rpm for 3 min, and the supernatant was colected and injected in the liquid chromatography of high resolution (HPLC). The results confirmed the presence of peramine in the larva's body of D. pallens. The insects that fed the experimental lines LQE-2 and LQE-6 were those where peramine was present in high proportion (0.205 and 0.199 ppm, respectively); while LQE-7 and LQE-3 obtained the lowest concentrations of the alkaloid (0.047 and 0.053 ppm, respectively). Peramine was not detected in the insects when the control cultivar Jumbo (E-) was tested. These results evidenced the storage and metabolism of peramine during consumption of the larvae. However, the effect of this alkaloid present in 'future ryegrass cultivars' (LQE-2 and LQE-6) on the performance and survival of blackworms must be studied and confirmed experimentally.

Keywords: blackworms, HPLC, alkaloid, pest

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Authors: Engy Shokry, Giancarlo La Marca, Maria Luisa Della Bona

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Newborn screening for Congenital Adrenal Hyperplasia (CAH) relies on quantification of 17α-hydroxyprogesterone using enzyme immunoassays. These assays, in spite of being rapid, readily available and easy to perform, its reliability was found questionable due to lack of selectivity and specificity resulting in large number of false-positives, consequently family anxiety and associated hospitalization costs. To improve specificity of conventional 17α-hydroxyprogesterone screening which may experience false transient elevation in preterm, low birth weight or acutely ill neonates, steroid profiling by LC-MS/MS as a second-tier test was implemented. Unlike the previously applied LC-MS/MS methods, with the disadvantage of requiring a relatively high number of blood drops. Since newborn screening tests are increasing, it is necessary to minimize the sample volume requirement to make the maximum use of blood samples collected on filter paper. The proposed new method requires just one 3.2 mm dried blood spot (DBS) punch. Extraction was done using methanol: water: formic acid (90:10:0.1, v/v/v) containing deuterium labelled internal standards. Extracts were evaporated and reconstituted in 10 % acetone in water. Column switching strategy for on-line sample clean-up was applied to improve the chromatographic run. The first separative step retained the investigated steroids and passed through the majority of high molecular weight impurities. After the valve switching, the investigated steroids are back flushed from the POROS® column onto the analytical column and separated using gradient elution. Found quantitation limits were 5, 10 and 50 nmol/L for 17α-hydroxyprogesterone, androstenedione and cortisol respectively with mean recoveries of between 98.31-103.24 % and intra-/ inter-assay CV% < 10 % except at LLOQ. The method was validated using standard addition calibration and isotope dilution strategies. Reference ranges were determined by analysing samples from 896 infants of various ages at the time of sample collection. The method was also applied on patients with confirmed CAH. Our method represents an attractive combination of low sample volume requirement, minimal sample preparation time without derivatization and quick chromatography (5 min). The three steroid profile and the concentration ratios (17OHP + androstenedione/cortisol) allowed better screening outcomes of CAH reducing false positives, associated costs and anxiety.

Keywords: congenital adrenal hyperplasia (CAH), 17α-hydroxyprogesterone, androstenedione, cortisol, LC-MS/MS

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Authors: Zachary Muthii Rukenya, James Mbaria, Peter Mbaabu, Kiama Stephen Gitahi, Ronald Onzago

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Aloe turkanensis is one of the widely used medicinal shrub and in Kenya the plant is mainly found in Baringo, Isiolo, Laikipia, Turkana and West Pokot Counties where it is used in ethno-medicine and ethno-veterinary medicine. The Turkana community uses the plant products to manage malaria, wounds, stomach ache, constipation, pain, skin infection, poultry diseases and retained afterbirth in cows. This evaluated the efficacy and safety of the plant obtained from Turkana County, Kenya. Preliminary data on the use of the plant in the county was collected through observation, photographing and interviews. A sample of the whole plant was harvested in Natira sublocation, in ex-Turkana west district in February 2012 after identification by Aloe-working group herbalists who voluntarily provided information on its medicinal uses. Botanical identification was done at Kenya Forest Research Centre in Karura where voucher specimen was deposited. Cold maceration using 70% methanol and distilled water was used for extraction. Bioassays were to determine the effects of the plant extracts on brine shrimp and selected bacterial and fungal cultures. The extracts were tested in-vitro activity against standard cultures of B. cereus (ATCC 11778), S. aureus (ATCC25923), P. aeroginosa (ATCC 27853), E. coli (ATCC 25922) and a human infections clinical isolate of C. albicans. The extracts of Aloe turkanensis inhibited the growth B. cereus (100-200 mg/ml), S. aureus (50-100 mg/ml), P. aeroginosa (200mg/ml), E. coli (400mg/ml) while C. albicans was not affected. The extracts also inhibited the growth of S. aureus and B. cereus with mean diameters of inhibition zones being 19.75±1 mm and 18.5±05 mm reapectively. Phytochemical screening showed the presence of alkaloids, tarpenoids, steroids, quinones, saponins and tannins in the plant extracts. The extract was found to be non-toxic at a concentration of 1000µg/ml with a 100% survival of Brine Shrimp larva. It was concluded that Aloe turkanensis growing the study area has metabolites that inhibit the growth of microorganisms and is however, there is need for further studies to validate the in-vivo bioactivity of the plant and more generate adequate toxicological data.to support conservation, value chain addition of its products and widespread use as a herbal remedy.

Keywords: Aloe turkanensis, bioactivity, cultivated, human infections

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Authors: Danish Laidin, Peter Gostomski, Aaron Marshall, Carlo Carere

Abstract:

Groundwater contamination of nitrate (NO3-) is becoming more prevalent in regions of intensive and extensive agricultural activities. Household nitrate removal involves using ion exchange membranes and reverse osmosis (RO) systems, whereas industrial nitrate removal may use organic carbon substrates (e.g. methanol) for heterotrophic microbial denitrification. However, these approaches both require high capital investment and operating costs. In this study, denitrification was demonstrated using bio-electrochemical systems (BESs) inoculated from sediments and microbial enrichment cultures. The BES reactors were operated continuously as microbial electrolytic cells (MECs) with a poised potential of -0.7V and -1.1V vs Ag/AgCl. Three parallel MECs were inoculated using hydrogen-driven denitrifying enrichments, stream sediments, and biofilm harvested from a denitrifying biotrickling filter, respectively. These reactors were continuously operated for over a year as various operating conditions were investigated to determine the optimal conditions for electroactive denitrification. The mass loading rate of nitrate was varied between 10 – 70 mg NO3-/d, and the maximum observed nitrate removal rate was 22 mg NO3- /(cm2∙d) with a current of 2.1 mA. For volumetric load experiments, the dilution rate of 1 mM NO3- feed was varied between 0.01 – 0.1 hr-1 to achieve a nitrate loading rate similar to the mass loading rate experiments. Under these conditions, the maximum rate of denitrification observed was 15.8 mg NO3- /(cm2∙d) with a current of 1.7mA. Hydrogen (H2) was supplied intermittently to investigate the hydrogenotrophic potential of the denitrifying biofilm electrodes. H2 supplementation at 0.1 mL/min resulted in an increase of nitrate removal from 0.3 mg NO3- /(cm2∙d) to 3.4 mg NO3- /(cm2∙d) in the hydrogenotrophically subcultured reactor but had no impact on the reactors which exhibited direct electron transfer properties. Results from this study depict the denitrification performance of the immobilized biofilm electrodes, either by direct electron transfer or hydrogen-driven denitrification, and the contribution of the planktonic cells present in the growth medium. Other results will include the microbial community analysis via 16s rDNA amplicon sequencing, varying the effect of poising cathodic potential from 0.7V to 1.3V vs Ag/AgCl, investigating the potential of using in-situ electrochemically produced hydrogen for autotrophic denitrification and adjusting the conductivity of the feed solution to mimic groundwater conditions. These findings highlight the overall performance of sediment inoculated MECs in removing nitrate and will be used for the future development of sustainable solutions for the treatment of nitrate polluted groundwater.

Keywords: bio-electrochemical systems, groundwater, electroactive denitrification, microbial electrolytic cell

Procedia PDF Downloads 58

Authors: Teruya Goto, Hokuto Chiba, Tatsuhiro Takahashi

Abstract:

Carbon fiber reinforced thermoplastic resin using short carbon fiber has been produced by melt mixing and the improvement of mechanical properties has been frequently reported up to now. One of the most frequently reported enhancement has been seen in carbon fiber / polypropylene (PP) composites by adding small amount of maleic anhydride grafted polypropylene (MA-g-PP) into PP matrix. However, the further enhancement of tensile strength and tensile modules has been expected for lightning the composite more. Our present research aims to improve the mechanical property by using a highly reactive monolayer polymer, which can react with both COOH of carbon fiber surface and maleic anhydride of MA-g-PP in the matrix, on carbon fiber for PP/CF composite. It has been known that oxazoline has much higher reactivity with COOH without catalysts, compared with amine group and alcohol OH group. However, oxazoline group has not been used for the interface. To achieve the purpose, poly-2-vinyl-2-oxazoline (Pvozo), having highly reactivity with COOH and maleic anhydride, has been originally synthesized through radical polymerization using 2-vinyl-2-oxazoline as a monomer, resulting in the Mw around 140,000. Monolayer Pvozo chemically reacted on CF was prepared in 1-methoxy-2-propanol solution of Pvozo by heating at 100oC for 3 hours. After this solution treatment, unreacted Pvozo was completely washed out by methanol, resulting the uniform formation of the monolayer Pvozo on CF. Monolayer Pvozo coated CF was melt mixed by with PP and a small amount of MA-g-PP for the preparation of the composite samples using a batch type melt mixer. With performing the tensile strength tests of the composites, the tensile strength of CF/MA-g-PP/PP showed 40% increase, compared to that of CF/PP. While, that of Pvozo coated CF/MA-g-PP/PP exhibited 80% increase, compared to that of CF/PP. To get deeper insight of the dramatic increase, the weight percentage of chemically grafted polymer based on CF was evaluated by dissolving and removing the matrix polymer by xylene using by thermos gravimetric analysis (TGA). The chemically grafted remained polymer was found to be 0.69wt% in CF/PP, 0.98wt% in CF/MA-g-PP/PP, 1.51wt% in Pvozo coated CF/MA-g-PP/PP, suggesting that monolayer Pvozo contributed to the increase of the grafted polymer amount. In addition, the very strong adhesion by Pvozo was confirmed by observing the fractured cross-sectional surface of the composite by scanning electron micrograph (SEM). As a conclusion, the effectiveness of a highly reactive monolayer Pvozo on CF for the enhancement of the mechanical properties of CF/PP composite was demonstrated, which can be interpreted by the clear evidence of the increase of the grafting polymer on CF.

Keywords: CFRTP, interface, oxazoline, polymer graft, mechanical property

Procedia PDF Downloads 195