Search results for: enzyme blend

Authors: Shameel Pervez, Saad Aziz Durrani, Ibatsam Khokhar

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Pectinase is an enzyme that breaks down pectin, a compound responsible for structural integrity of the plant. Pectin is difficult to break down mechanically and the cost is very high, that is why many industries including food industries use pectinase enzyme produced by microbes for pectin breakdown. Apple (Malus domestica) is an important fruit in terms of market value. Every year, millions of apples are wasted due to post-harvest rot caused by fungi. Fungi are natural decomposers of our ecosystem and are infamous for post-harvest rot of apple fruit but at the same time they are prized for their high production of valuable extracellular enzymes such as pectinase. In this study, fungi belonging to different genus were isolated from rotten apples. Rotten samples of apple were picked from different markets of Lahore. After surface sterilization, the rotten parts were cut into small pieces and placed onto MEA media plates for three days. Afterwards, distinct colonies were picked and purified by sub-culturing. The isolates were identified to genus level through the study of basic colony morphology and microscopic features. The isolates were then subjected to screening for pectinase activity on MS media to compare pectinase production and were then subsequently tested for pathogenic activity through wound suspension method to evaluate the pathogenic activity of isolates in comparison with their pectinolytic activity. A total of twelve fungal strains were isolates from rotten apples. They were belonging to genus Penicillium, Alternaria, Paecilomyces and Rhizopus. Upon screening for pectinolytic activity, isolates Pen 1, Pen 4, and Rz showed high pectinolytic activity and were further subjected to DNA isolation and partial sequencing for species identification. The results of partial sequencing were combined with in-depth study of morphological features revealing Pen 1 as Penicillium janthinellum, Pen 4 as Penicillium griseofulvum, and Rz as Rhizopus microsporus. Pathogenic activity of all twelve isolates was evaluated. Penicillium spp. were highly pathogenic and destructive and same was the case with Paecilomyces sp. and Rhizopus sp. However, Alternaria spp. were found to be more consistent in their pathogenic activity, on all types of apples.

Keywords: apple, pectinase, fungal pathogens, penicillium, rhizopus

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Authors: Navpreet Kaur, Himkusha Thakur, Nirmal Prabhakar

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One of the most publicized and controversial issue in crop production is the use of agrichemicals- also known as pesticides. This is evident in many reports that Organophosphate (OP) insecticides, among the broad range of pesticides are mainly involved in acute and chronic poisoning cases. Therefore, detection of OPs is very necessary for health protection, food and environmental safety. In our study, a nanocomposite of poly (3,4 ethylenedioxythiophene) (PEDOT) and multi-walled carbon nanotubes (MWCNTs) has been deposited electrochemically onto the surface of fluorine doped tin oxide sheets (FTO) for the analysis of malathion OP. The -COOH functionalization of MWCNTs has been done for the covalent binding with amino groups of AChE enzyme. The use of PEDOT-MWCNT films exhibited an excellent conductivity, enables fast transfer kinetics and provided a favourable biocompatible microenvironment for AChE, for the significant malathion OP detection. The prepared PEDOT-MWCNT/FTO and AChE/PEDOT-MWCNT/FTO nano-biosensors were characterized by Fourier transform infrared spectrometry (FTIR), Field emission-scanning electron microscopy (FE-SEM) and electrochemical studies. Electrochemical studies were done using Cyclic Voltammetry (CV) or Differential Pulse Voltammetry (DPV) and Electrochemical Impedance Spectroscopy (EIS). Various optimization studies were done for different parameters including pH (7.5), AChE concentration (50 mU), substrate concentration (0.3 mM) and inhibition time (10 min). The detection limit for malathion OP was calculated to be 1 fM within the linear range 1 fM to 1 µM. The activity of inhibited AChE enzyme was restored to 98% of its original value by 2-pyridine aldoxime methiodide (2-PAM) (5 mM) treatment for 11 min. The oxime 2-PAM is able to remove malathion from the active site of AChE by means of trans-esterification reaction. The storage stability and reusability of the prepared nano-biosensor is observed to be 30 days and seven times, respectively. The application of the developed nano-biosensor has also been evaluated for spiked lettuce sample. Recoveries of malathion from the spiked lettuce sample ranged between 96-98%. The low detection limit obtained by the developed nano-biosensor made them reliable, sensitive and a low cost process.

Keywords: PEDOT-MWCNT, malathion, organophosphates, acetylcholinesterase, nano-biosensor, oxime (2-PAM)

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Authors: Ozlem Oral, Emre Taskin, Aysel Yuce, Serap Dokmeci, Devrim Gozuacik

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Autophagy is an evolutionary conserved lysosome-dependent catabolic pathway, responsible for the degradation of long-lived proteins, abnormal aggregates and damaged organelles which cannot be degraded by the ubiquitin-proteasome system. Lysosomes degrade the substrates through the activity of lysosomal hydrolases and lysosomal membrane-bound proteins. Mutations in the coding region of these proteins cause malfunctional lysosomes, which contributes to the pathogenesis of lysosomal storage diseases. Gaucher disease is a lysosomal storage disease resulting from the mutation of a lysosomal membrane-associated glycoprotein called glucocerebrosidase and its cofactor saposin C. The disease leads to intracellular accumulation of glucosylceramide and other glycolipids. Because of the essential role of lysosomes in autophagic degradation, Gaucher disease may directly be linked to this pathway. In this study, we investigated the expression of autophagy and/or lysosome-related genes and proteins in fibroblast cells isolated from patients with different mutations. We carried out confocal microscopy analysis and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. We also evaluated lysosomal pH by active lysosome staining and lysosomal enzyme activity. Beside lysosomes, we also performed proteasomal activity and cell death analysis in patient samples. Our data showed significant attenuation in the expression of key autophagy-related genes and accumulation of their proteins in mutant cells. We found decreased the ability of autophagosomes to fuse with lysosomes, associated with elevated lysosomal pH and reduced lysosomal enzyme activity. Proteasomal degradation and cell death analysis showed reduced proteolytic activity of the proteasome, which consequently leads to increased susceptibility to cell death. Our data indicate that the major degradation pathways are affected by multifunctional lysosomes in mutant patient cells and may underlie in the mechanism of clinical severity of Gaucher patients. (This project is supported by TUBITAK-3501-National Young Researchers Career Development Program, Project No: 112T130).

Keywords: autophagy, Gaucher's disease, glucocerebrosidase, mutant fibroblasts

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Authors: Abdul Ghaffar Memon, Xiao Hong Zhou, Yunpeng Xing, Ruoyu Wang, Miao He

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Due to deleterious environmental and health effects of the Hg²⁺ ions, various online, detection methods apart from the traditional analytical tools have been developed by researchers. Biosensors especially, label, label-free, colorimetric and optical sensors have advanced with sensitive detection. However, there remains a gap of ultrasensitive quantification as noise interact significantly especially in the AuNP based label-free colorimetry. This study reported an amplification strategy using Exo-III enzyme for target recycling of Hg²⁺ ions in a T-rich hairpin loop metallobase label-free colorimetric nanosensor with an improved sensitivity using unmodified gold nanoparticles (uGNPs) as an indicator. The two T-rich metallobase hairpin loop structures as 5’- CTT TCA TAC ATA GAA AAT GTA TGT TTG -3 (HgS1), and 5’- GGC TTT GAG CGC TAA GAA A TA GCG CTC TTT G -3’ (HgS2) were tested in the study. The thermodynamic properties of HgS1 and HgS2 were calculated using online tools (http://biophysics.idtdna.com/cgi-bin/meltCalculator.cgi). The lab scale synthesized uGNPs were utilized in the analysis. The DNA sequence had T-rich bases on both tails end, which in the presence of Hg²⁺ forms a T-Hg²⁺-T mismatch, promoting the formation of dsDNA. Later, the Exo-III incubation enable the enzyme to cleave stepwise mononucleotides from the 3’ end until the structure become single-stranded. These ssDNA fragments then adsorb on the surface of AuNPs in their presence and protect AuNPs from the induced salt aggregation. The visible change in color from blue (aggregation stage in the absence of Hg²⁺) and pink (dispersion state in the presence of Hg²⁺ and adsorption of ssDNA fragments) can be observed and analyzed through UV spectrometry. An ultrasensitive quantitative nanosensor employing Exo-III assisted target recycling of mercury ions through label-free colorimetry with nanomolar detection using uGNPs have been achieved and is further under the optimization to achieve picomolar range by avoiding the influence of the environmental matrix. The proposed strategy will supplement in the direction of uGNP based ultrasensitive, rapid, onsite, label-free colorimetric detection.

Keywords: colorimetric, Exo-III, gold nanoparticles, Hg²⁺ detection, label-free, signal amplification

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Authors: Nisha Singh, Munish Puri, Collin Barrow, Deepak Tuli, Anshu S. Mathur

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The biological conversion of lignocellulosic biomass to ethanol is a promising strategy to solve the present global crisis of exhausting fossil fuels. The existing bioethanol production technologies have cost constraints due to the involvement of mandate pretreatment and extensive enzyme production steps. A unique process configuration known as consolidated bioprocessing (CBP) is believed to be a potential cost-effective process due to its efficient integration of enzyme production, saccharification, and fermentation into one step. Due to several favorable reasons like single step conversion, no need of adding exogenous enzymes and facilitated product recovery, CBP has gained the attention of researchers worldwide. However, there are several technical and economic barriers which need to be overcome for making consolidated bioprocessing a commercially viable process. Finding a natural candidate CBP organism is critically important and thermophilic anaerobes are preferred microorganisms. The thermophilic anaerobes that can represent CBP mainly belong to genus Clostridium, Caldicellulosiruptor, Thermoanaerobacter, Thermoanaero bacterium, and Geobacillus etc. Amongst them, Clostridium thermocellum has received increased attention as a high utility CBP candidate due to its highest growth rate on crystalline cellulose, the presence of highly efficient cellulosome system and ability to produce ethanol directly from cellulose. Recently with the availability of genetic and molecular tools aiding the metabolic engineering of Clostridium thermocellum have further facilitated the viability of commercial CBP process. With this view, we have specifically screened cellulolytic and xylanolytic thermophilic anaerobic ethanol producing bacteria, from unexplored hot spring/s in India. One of the isolates is a potential CBP organism identified as a new strain of Clostridium thermocellum. This strain has shown superior avicel and xylan degradation under unoptimized conditions compared to reported wild type strains of Clostridium thermocellum and produced more than 50 mM ethanol in 72 hours from 1 % avicel at 60°C. Besides, this strain shows good ethanol tolerance and growth on both hexose and pentose sugars. Hence, with further optimization this new strain could be developed as a potential CBP microbe.

Keywords: Clostridium thermocellum, consolidated bioprocessing, ethanol, thermophilic anaerobes

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Authors: Satwik Majumder, Dongyun Jung, Jennifer Ronholm, Saji George

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Bovine mastitis is the most common infectious disease in dairy cattle, with major economic implications for the dairy industry worldwide. Continuous monitoring for the emergence of antimicrobial resistance (AMR) among bacterial isolates from dairy farms is vital not only for animal husbandry but also for public health. In this study, the prevalence of AMR in 113 Escherichia coli isolates from cases of bovine clinical mastitis in Canada was investigated. Kirby-Bauer disk diffusion test with 18 antibiotics and microdilution method with three heavy metals (copper, zinc, and silver) was performed to determine the antibiotic and heavy-metal susceptibility. Resistant strains were assessed for efflux and ß-lactamase activities besides assessing biofilm formation and hemolysis. Whole-genome sequences for each of the isolates were examined to detect the presence of genes corresponding to the observed AMR and virulence factors. Phenotypic analysis revealed that 32 isolates were resistant to one or more antibiotics, and 107 showed resistance against at least one heavy metal. Quinolones and silver were the most efficient against the tested isolates. Among the AMR isolates, AcrAB-TolC efflux activity and ß-lactamase enzyme activities were detected in 13 and 14 isolates, respectively. All isolates produced biofilm but with different capacities, and 33 isolates showed α-hemolysin activity. A positive correlation (Pearson r = +0.89) between efflux pump activity and quantity of biofilm was observed. Genes associated with aggregation, adhesion, cyclic di-GMP, quorum sensing were detected in the AMR isolates, corroborating phenotype observations. This investigation showed the prevalence of AMR in E. coli isolates from bovine clinical mastitis. The results also suggest the inadequacy of antimicrobials with a single mode of action to curtail AMR bacteria with multiple mechanisms of resistance and virulence factors. Therefore, it calls for combinatorial therapy for the effective management of AMR infections in dairy farms and combats its potential transmission to the food supply chain through milk and dairy products.

Keywords: antimicrobial resistance, E. coli, bovine mastitis, antibiotics, heavy-metals, efflux pump, ß-lactamase enzyme, biofilm, whole-genome sequencing

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Authors: Caroline M. Spagnol, Vera L. B. Isaac, Marcos A. Corrêa, Hérida R. N. Salgado

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Phenolic compounds are abundant in the Brazilian plant kingdom and they are part of a large and complex group of organic substances. Cinnamic acids are part of this group of organic compounds, and caffeic acid (CA) is one of its representatives. Antioxidants are compounds which act as free radical scavengers and, in other cases, such as metal chelators, both in the initiation stage and the propagation of oxidative process. The tyrosinase, polyphenol oxidase, is an enzyme that acts at various stages of melanin biosynthesis within the melanocytes and is considered a key molecule in this process. Some phenolic compounds exhibit inhibitory effects on melanogenesis by inhibiting the tyrosinase enzymatic activity and therefore has been the subject of studies. However, few studies have reported the effectiveness of these products and their safety. Objectives: To assess the inhibitory activity of tyrosinase, the antioxidant activity of CA and its cytotoxic potential. The method to evaluate the inhibitory activity of tyrosinase aims to assess the reduction transformation of L-dopa into dopaquinone reactions catalyzed by the enzyme. For evaluating the antioxidant activity was used the analytical methodology of DPPH radical inhibition. The cytotoxicity evaluation was carried out using the MTT method (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide), a colorimetric assay which determines the amount of insoluble violet crystals formed by the reduction of MTT in the mitochondria of living cells. Based on the results obtained during the study, CA has low activity as a depigmenting agent. However, it is a more potent antioxidant than ascorbic acid (AA), since a lower amount of CA is sufficient to inhibit 50% of DPPH radical. The results are promising since CA concentration that promoted 50% toxicity in HepG2 cells (IC50=781.8 μg/mL) is approximately 330 to 400 times greater than the concentration required to inhibit 50% of DPPH (IC50 DPPH= 2.39 μg/mL) and ABTS (IC50 ABTS= 1.96 μg/mL) radicals scavenging activity, respectively. The maximum concentration of caffeic acid tested (1140 mg /mL) did not reach 50% of cell death in HaCat cells. Thus, it was concluded that the caffeic acid does not cause toxicity in HepG2 and HaCat cells in the concentrations required to promote antioxidant activity in vitro, and it can be applied in topical products.

Keywords: caffeic acid, antioxidant, cytotoxicity, cosmetic

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Authors: Jalpit B. Prajapati, Ketankumar G. Patel

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Palm oil may be employed in diesel engine as an alternative fuel. Biofuel has so far been backed by government policies in the quest for low carbon fuel in the near future and promises to ensure energy security through partially replacing fossil fuels. This paper presents an experimental investigation of performance and emission characteristics by using palm oil in diesel engine. The properties of palm oil can be compared favorably with the characteristics required for internal combustion engine fuels especially diesel engine. Experiments will be performed for fixed compression ratio i.e. 18 using biodiesel-diesel blends i.e. B0, B10, B20, B30, B40, B50 with load variation from no load to full load and compared with base cases i.e. engine using diesel as a fuel. The parameters studied in performance characteristics are brake power, brake specific fuel consumption and brake thermal efficiency, in emission characteristics are carbon monoxide, unburnt hydrocarbons and nitrogen oxide. After experimental results B20 (20% palm oil and 80% diesel) is best in performance, but NOx formation is little higher in B20.

Keywords: palm biodiesel, performance, emission, diesel-biodiesel blend

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Authors: N. Schneider, M. Bruchhage, M. Hartweg, G. Mutungi, J. O Regan, S. Deoni

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Observational neuroimaging studies suggest differences between breast-fed and formula-fed infants in developmental myelination, a key brain process for learning and cognitive development. However, the possible effects of a nutrient formulation on myelin development in healthy term infants in an intervention study have not been investigated. Objective was, therefore, to investigate the efficacy of a nutrient formulation with higher levels of myelin-relevant nutrients as compared to a control formulation with lower levels of the same nutrients on brain myelination and cognitive development in the first 6 months of life. The study is an ongoing randomized, controlled, double-blind, two-center, parallel-group clinical trial with a nonrandomized, non-blinded arm of exclusively breastfed infants. The current findings result from a staged statistical analysis at 6 months; the recruitment and intervention period has been completed for all participants. Follow-up visits at 12, 18 and 24 months are still ongoing. N= 81 enrolled full term, neurotypical infants of both sexes were randomized into either the investigational (N= 42) or the control group (N= 39), and N= 108 children in the breast-fed arm served as a natural reference group. The effect of a blend of docosahexaenoic acid, arachidonic acid, iron, vitamin B12, folic acid as well as sphingomyelin from a uniquely proceed whey protein concentrate enriched in alpha-lactalbumin and phospholipids in an infant nutrition product matrix was investigated. The main outcomes for the staged statistical analyses at 6 months included brain myelination measures derived from MRI. Additional outcomes were brain volume, cognitive development and safety. The full analyses set at 6 months comprised N= 66 infants. Higher levels of myelin-relevant nutrients compared to lower levels resulted in significant differences in myelin structure, volume, and rate of myelination as early as 3 and 6 months of life. The cross-sectional change of means between groups for whole-brain myelin volume was 8.4% for investigational versus control formulation (3.5% versus the breastfeeding reference) group at 3 months and increased to 36.4% for investigational versus control formulation (14.1% versus breastfeeding reference) at 6 months. No statistically significant differences were detected for early cognition scores. Safety findings were largely similar across groups. This is the first pediatric nutritional neuroimaging study demonstrating the efficacy of a myelin nutrient blend on developmental myelination in well-nourished term infants. Myelination is a critical process in learning and development. The effects were demonstrated across the brain, particularly in temporal and parietal regions, known to be functionally involved in sensory, motor and language skills. These first results add to the field of nutritional neuroscience by demonstrating early life nutrition benefits for brain architecture which may be foundational for later cognitive and behavioral outcomes. ClinicalTrials.gov Identifier: NCT03111927 (Infant Nutrition and Brain Development - Full-Text View - ClinicalTrials.gov).

Keywords: brain development, infant nutrition, MRI, myelination

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Authors: Pampita Chakraborty, Sukumar Mukherjee

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Diabetes Mellitus (DM) is commonly encountered metabolic disorder in clinical practice. An estimated 25 percent of DM patients develop foot problems. Foot ulceration and infection are one of the major causes of morbidity, hospitalization or even amputation. Objective: To isolate and identify bacterial pathogens in Diabetic Foot Ulcer (DFU) and to observe its antimicrobial sensitivity pattern. Methodology: A prospective study was conducted for a period of 9 months at the Department of Microbiology, GD Hospital & Diabetes Institute, Kolkata. 75 DFU patients were recruited in the study. Specimens for microbiological studies obtained from ulcer base were examined as gram stained smear and was cultured aerobically on Nutrient agar, Blood agar and MacConkey agar plates. Antimicrobial sensitivity test was performed by disc diffusion techniques according to CLSI guidelines. Result: In this study out of 75cases, 73% (55/75) were male and 27% (20/75) were females with mean (SD) age of 51.11(±10) years. Out of 75 pus cultures, 63(84%) showed growth of microorganism making total of 81 bacterial isolates with 71.42% of monomicrobial infection and 28.57% of polymicrobial infection. Out of 81 isolates 53(65.43%) were gram negative and 21(25.92%) were gram positive. E.coli was relatively common isolate 21(26%) followed by Staphylococcus aureus 15(18.5%), Klebsiella pneumonia 14(17.28%), Pseudomonas aeruginosa 12 (14.81%), Proteus spp. 3 (3.70%), and Enterococcus faecalis 6 (7.40%). 75% of Gram-negative microorganism were extended Beta-lactamase enzyme (ESBL) producer and around 20 % of Klebsiella and Proteus spp. were carbapenemase enzyme producer. Among Gram positive, around 50% of S.aureus was MRSA, sensitive only to Vancomycin, Teicoplanin & Linezolid. Conclusion: More prevalence of monomicrobial gram-negative bacteria than gram-positive bacteria in DFU was observed. This study emphasizes that Beta-Lactam group of antibiotics should not be the empirical treatment of choice for Gram-negative isolates; instead alternatives like Carbapenems, Amikacin could be a better option. On the other hand, Vancomycin and Linezolid are preferred for most of the infection with gram-positive aerobes. Continuous surveillance of resistant bacteria is required for empiric therapy.

Keywords: antibiotic resistant, antimicrobial susceptibility, diabetic foot ulcer, surveillance

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Authors: Epameinondas Xanthakis, Erik Kaunisto, Alain Le-Bail, Lilia Ahrné

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Fruits and vegetables are characterized as perishable food matrices due to their short shelf life as several deterioration mechanisms are being involved. Prior to the common preservation methods like freezing or canning, fruits and vegetables are being blanched in order to inactivate deteriorative enzymes. Both conventional blanching pretreatments and conventional freezing methods hide drawbacks behind their beneficial impacts on the preservation of those matrices. Conventional blanching methods may require longer processing times, leaching of minerals and nutrients due to the contact with the warm water which in turn leads to effluent production with large BOD. An important issue of freezing technologies is the size of the formed ice crystals which is also critical for the final quality of the frozen food as it can cause irreversible damage to the cellular structure and subsequently to degrade the texture and the colour of the product. Herein, the developed microwave blanching methodology and the results regarding quality aspects and enzyme inactivation will be presented. Moreover, heat transfer phenomena, mass balance, temperature distribution, and enzyme inactivation (such as Pectin Methyl Esterase and Ascorbic Acid Oxidase) of our microwave blanching approach will be evaluated based on measurements and computer modelling. The present work is part of the COLDμWAVE project which aims to the development of an innovative environmentally sustainable process for blanching and freezing of fruits and vegetables with improved textural and nutritional quality. In this context, COLDµWAVE will develop tailored equipment for MW blanching of vegetables that has very high energy efficiency and no water consumption. Furthermore, the next steps of this project regarding the development of innovative pathways in MW assisted freezing to improve the quality of frozen vegetables, by exploring in depth previous results acquired by the authors, will be presented. The application of MW assisted freezing process on fruits and vegetables it is expected to lead to improved quality characteristics compared to the conventional freezing. Acknowledgments: COLDμWAVE has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grand agreement No 660067.

Keywords: blanching, freezing, fruits, microwave blanching, microwave

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Authors: Miramar Damanhouri

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This study examines the relationship between language use in computer-mediated communication and the social identity of the user. The data were collected by surveying 298 Saudi bilingual speakers who are familiar with Arabizi, a blend of Latin characters and Arabic numerals to transliterate Arabic sounds, and then analyzed quantitatively by running tests for statistical confidence in order to determine differences in perceptions between young adults (ages 15-25 years) and middle-aged adults (ages 26-50 years). According to the findings of this study, English is the dominant language among most of the young adults surveyed, and when they do use Arabic, they use Arabizi because of its flexibility, compatibility with modern technology, and its acceptance among people of their age and sociocultural backgrounds. On the other hand, most middle-aged adults surveyed here tend to use Arabic, as they believe that they should show their loyalty to their origin. The results of the study demonstrate a mutual relationship between language use in computer-mediated communication and the user’s social identity, as language is used both to reflect and construct that identity.

Keywords: Arabizi, computer mediated communication, digital communication, language use

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Authors: M. Mourad, K. Abdelgawwad

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This experimental investigation aimed to determine the influence of using different types of alcohol and gasoline blends such as ethanol - butanol - propanol on the performance of spark ignition engine. The experimental work studied the effect of various fuel blends such as ethanol – butanol/gasoline and propanol/gasoline with two rates of 15% and 20%, at different operating conditions (engine speed and loads), on engine performance emission characteristics. Laboratory experiments are carried out on a four-cylinder spark ignition (SI) engine. In this practical study, all considerations and precautions are taken into account to ensure the quality and accuracy of practical experiments and different measurements. The results show that the performance of the engine improved significantly in the case of ethanol/butanol-gasoline blends. The results also indicated that the engine emitted pollutants such as CO, hydrocarbon (HC) for alcohol fuel blends compared to base gasoline NOx emission increased for different fuel blends either ethanol/butanol-gasoline or propanol-gasoline fuel blend.

Keywords: gasoline engine, performance, emission, fuel blends

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Authors: Mannar R. Maurya, Bithika Sarkar, Fernando Avecilla

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Peroxidase activity is possibly successfully used for different industrial processes in medicine, chemical industry, food processing and agriculture. However, they bear some intrinsic drawback associated with denaturation by proteases, their special storage requisite and cost factor also. Now a day’s artificial enzyme mimics are becoming a research interest because of their significant applications over conventional organic enzymes for ease of their preparation, low price and good stability in activity and overcome the drawbacks of natural enzymes e.g serine proteases. At present, a large number of artificial enzymes have been synthesized by assimilating a catalytic center into a variety of schiff base complexes, ligand-anchoring, supramolecular complexes, hematin, porphyrin, nanoparticles to mimic natural enzymes. Although in recent years a several number of vanadium complexes have been reported by a continuing increase in interest in bioinorganic chemistry. To our best of knowledge, the investigation of artificial enzyme mimics of vanadium complexes is very less explored. Recently, our group has reported synthetic vanadium schiff base complexes capable of mimicking peroxidases. Herein, we have synthesized monoidovanadium(IV) and dioxidovanadium(V) complexes of pyrazoleone derivateis ( extensively studied on account of their broad range of pharmacological appication). All these complexes are characterized by various spectroscopic techniques like FT-IR, UV-Visible, NMR (1H, 13C and 51V), Elemental analysis, thermal studies and single crystal analysis. The peroxidase mimic activity has been studied towards oxidation of pyrogallol to purpurogallin with hydrogen peroxide at pH 7 followed by measuring kinetic parameters. The Michaelis-Menten behavior shows an excellent catalytic activity over its natural counterparts, e.g. V-HPO and HRP. The obtained kinetic parameters (Vmax, Kcat) were also compared with peroxidase and haloperoxidase enzymes making it a promising mimic of peroxidase catalyst. Also, the catalytic activity has been studied towards the oxidation of 1-phenylethanol in presence of H2O2 as an oxidant. Various parameters such as amount of catalyst and oxidant, reaction time, reaction temperature and solvent have been taken into consideration to get maximum oxidative products of 1-phenylethanol.

Keywords: oxovanadium(IV)/dioxidovanadium(V) complexes, NMR spectroscopy, Crystal structure, peroxidase mimic activity towards oxidation of pyrogallol, Oxidation of 1-phenylethanol

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Authors: Debamitra Chakravorty, Pratap K. Parida

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Cold-adapted proteases enhance wash performance in low-temperature laundry resulting in a reduction in energy consumption and wear of textiles and are also used in the dehairing process in leather industries. Unfortunately, the possible drawbacks of using cold-adapted proteases are their instability at higher temperatures. Therefore, proteases with broad temperature stability are required. Unfortunately, wild-type cold-adapted proteases exhibit instability at higher temperatures and thus have low shelf lives. Therefore, attempts to engineer cold-adapted proteases by protein engineering were made previously by directed evolution and random mutagenesis. The lacuna is the time, capital, and labour involved to obtain these variants are very demanding and challenging. Therefore, rational engineering for cold stability without compromising an enzyme's optimum pH and temperature for activity is the current requirement. In this work, mutations were rationally designed with the aid of high throughput computational methodology of network analysis, evolutionary conservation scores, and molecular dynamics simulations for Savinase from Bacillus lentus with the intention of rendering the mutants cold stable without affecting their temperature and pH optimum for activity. Further, an attempt was made to incorporate a mutation in the most stable mutant rationally obtained by this method to introduce oxidative stability in the mutant. Such enzymes are desired in detergents with bleaching agents. In silico analysis by performing 300 ns molecular dynamics simulations at 5 different temperatures revealed that these three mutants were found to be better in cold stability compared to the wild type Savinase from Bacillus lentus. Conclusively, this work shows that cold adaptation without losing optimum temperature and pH stability and additionally stability from oxidative damage can be rationally designed by in silico enzyme engineering. The key findings of this work were first, the in silico data of H5 (cold stable savinase) used as a control in this work, corroborated with its reported wet lab temperature stability data. Secondly, three cold stable mutants of Savinase from Bacillus lentus were rationally identified. Lastly, a mutation which will stabilize savinase against oxidative damage was additionally identified.

Keywords: cold stability, molecular dynamics simulations, protein engineering, rational design

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Authors: Abubakar Umar Birnin-yauri

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Plastic wastes arising from Polyethylene (PE)-based materials are increasingly becoming environmental problem, this is owed to the fact that these PE waste materials will only decompose over hundreds, or even thousands of years, during which they cause serious environmental problems. In this research, Polymer blends prepared from PE, Cashewnut flour (CNF) and Gum Arabic (GA) were studied in order to assay their biodegradation potentials via composting method. Different sample formulations were made i.e., X1= (70% PE, 25% CNF and 5% GA, X2= (70% PE, 20% CNF and 10% GA), X3= (70% PE, 15% CNF and 15% GA), X4 = (70% PE, 10% CNF and 20% GA) and X5 = (70% PE, 5% CNF and 25% GA) respectively. The results obtained showed that X1 recorded weight loss of 9.89% of its original weight after the first 20 days and 37.45% after 100 day, and X2 lost 12.67 % after the first 20 days and 42.56% after 100day, sample X5 experienced the greatest weight lost in the two methods adopted which are 52.9% and 57.89%. Instrumental analysis such as Fourier Transform Infrared Spectroscopy, Thermogravimetric analysis and Scanning electron microscopy were performed on the polymer blends before and after biodegradation. The study revealed that the biodegradation of the polymer blends is influenced by the contents of both the CNF and GA added into the blends.

Keywords: polyethylene, cashewnut, gum Arabic, biodegradation, blend, environment

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Authors: Omaima A. Sharaf, Wafaa M. Abd El-Rahim, Hassan Moawad, Michael J. Sadowsky

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Textile industry is one of the important industries in the worldwide. It is known that the eco-friendly industrial and agricultural activities are significant for socio-economic stability of all countries. The absence of appropriate industrial waste water treatments is essential barrier for sustainable development in food and agricultural sectors especially in developing country like Egypt. Thus, the development of enzymatic bioremediation technology for textile dye removal will enhance the collaboration between scientists who develop the technology and industry where this technology will be implemented towards the safe disposal of the textile dye wastes. Highly efficient microorganisms are of most importance in developing and using highly effective biological treatment processes. Bacterial degradation of azo dyes is generally initiated by an enzymatic step that involves cleavage of azo linkages, usually with the aid of an azoreductase as electron donor. Thus, expanding the spectrum of microorganisms with high enzymatic activities as azoreductases and discovering novel azo-dye degrading enzymes, with enhanced stability and superior catalytic properties, are necessary for many environmental and industrial applications. Consequently, the use of molecular tools has become increasingly integrated into the understanding of enzyme properties and characterization. Researchers have utilized a gene cloning and expression methods as a tool to produce recombinant protein for decolorizing dyes more efficiently. Thus, presumptive evidence for the presence of genes encoding azoreductases in the genomes of selected local, and most potent azo-degrading strains were obtained by using specific oligonucleotides primers. These potent strains have been isolated from textile industrial wastewater in Egypt and identified using 16S rRNA sequence analysis as 'Lysinibacillus macrolidesB8, Brevibacillus parabrevisB11, Bacillus coagulansB7, and B. cereusB5'. PCR products of two full length genes designated as (AZO1;621bp and AZO2;534bp) were detected. BLASTx results indicated that AZO1 gene was corresponding to predicted azoreductase from of Bacillus sp. ABP14, complete genome, multispecies azoreductase [Bacillus], It was submitted to the gene bank by an accession no., BankIt2085371 AZO1 MG923210 (621bp; 207 amino acids). AZO1 was generated from the DNA of our identified strains Lysinibacillus macrolidesB8. On the other hand, AZO2 gene was corresponding to a predicted azoreductase from Bacillus cereus strain S2-8. Gene bank accession no. was BankIt2085839 AZO2 MG932081 (534bp;178 amino acids) and it was amplified from our Bacillus coagulansB7. Both genes were successfully cloned into pCR2.1TOPO (Invitrogen) and in pET28b+ vectors, then they transformed into E. coli DH5α and BL21(DE3) cells for heterologous expression studies. Our recombinant azoreductases (AZO1&AZO2) exhibited potential enzyme activity and efficiently decolorized an azo dye (Direct violet). They exhibited pH stability between 6 and 8 with optimum temperature up to 60°C and 37 °C after induction by 1mM and 1.5mM IPTG, for both AZO1 &AZO2, respectively. These results suggested that further optimization and purification of these recombinant proteins by using different heterologous expression systems will give great potential for the sustainable utilization of these recombinant enzymes in several industrial applications especially in wastewater treatments.

Keywords: azoreductases, decolorization, enzyme activity, gene cloning and expression

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Authors: Rita Riekstina-Dolge, Zanda Kruma, Fredijs Dimins, Inta Krasnova, Daina Karklina

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Apple juice is the main raw material for cider production. In this study apple juices obtained from 14 dessert and crab apples grown in Latvia were investigated. For all samples total phenolic compounds, tannins and individual phenolic compounds content were determined. The total phenolic content of different variety apple juices ranged from 650mg L-1 to 4265mg L-1. Chlorogenic acid is the predominant phenolic compound in all juice samples and ranged from 143.99mg L-1 in ‘Quaker Beauty’ apple juice to 617.66mg L-1 in ‘Kerr’ juice. Some dessert and crab apple juices have similar phenolic composition, but in several varieties such as ‘Cornelie’, ‘Hyslop’ and ‘Riku’ it was significantly higher. For cider production it is better to blend different kinds of apple juices including apples rich in high phenol content ('Rick', 'Cornelie') and also, for successful fermentation, apples rich in sugars and soluble solids content should be used in blends.

Keywords: apple juice, phenolic compounds, hierarchical cluster analysis, cider production

Procedia PDF Downloads 408

Authors: S. Badrinarayanan, R. Vimal, H. Sivaraman, P. Deepak, R. Vignesh Kumar, A. Ponshanmugakumar

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In the present study, the flammability properties of banana fibre reinforced epoxy/ sodium bromate blended composites are studied. Two sets of composite material were prepared, one formed by blending sodium bromate with epoxy matrix and other with neat epoxy matrix. Epoxy resin was blended with various weight fractions of sodium bromate, 4%, 8% and 12%. The composite made with plain epoxy matrix was used as the standard reference material. The mechanical tests, heat deflection tests and flammability tests were carried out on all the composite samples. Flammability test shows the improved flammability properties of the sodium bromated banana-epoxy composite. The modification in flammability properties of the composites by the addition of sodium bromate results in the reduced mechanical properties. The fractured surfaces under various mechanical testing were analysed using morphological analysis done using scanning electron microscope.

Keywords: banana fibres, epoxy resin, sodium bromate, flammability test, heat deflection

Procedia PDF Downloads 277

Authors: Jayisha Das Jaya, Nafis As Sami, Nazia Jahan, Tamanna Jerin, Mohammed Russedul Islam

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This paper contains continuous research which helps to analyze polypropylene modified bituminous mix and its impact characteristics with respect to original bitumen. Three percentages of polypropylene varying from (1-3) % of the weight of bitumen have been used to alter bitumen’s performance. The temperature of 170°C has been maintained during the blending of polypropylene with bitumen. It was performed by a wet process as it has certain advantages over the dry process. A rough estimate of 210 rpm rotation speed was set to prepare the blend in a mixer for 30 minutes producing homogeneous mixture. The blended mix shows a change in physical properties in comparison with the original bitumen content. Modification shows that for a 1% increment of polypropylene, softening point increases by 1 degree, penetration values decrease gradually to 55.6, 54, 52.5, ductility values decrease gradually to 87,76, 63 and specific gravity remains the same. Then Marshall mix design is performed with 60/70 penetration grade bitumen contents varying from (4-6) % with .5% intervals. Marshall stability and flow test results indicate the increase in stability and decrease in flow.

Keywords: bitumen, marshall, polypropylene, temperature

Procedia PDF Downloads 214

Authors: Boris Nemzer, Tania Reyes-Izquierdo, Zbigniew Pietrzkowski

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Glucosinolate is a family of glucosides that can be found in a family of brassica vegetables. Upon the damage of the plant, glucosinolate breakdown by an internal enzyme myrosinase (thioglucosidase; EC 3.2.3.1) into isothiocyanates, such as sulforaphane. Sulforaphane is formed by glucoraphanin cleaving the sugar off by myrosinase and rearranged. Sulforaphane nitrile is formed in the same reaction as sulforaphane with the active of epithiospecifier protein (ESP). Most common food processing procedure would break the plant and mix the glucoraphanin and myrosinase together, and the formed sulforaphane would be further degraded. The purpose of this study is to understand the glucoraphanin/sulforaphane and the myrosinase activity of broccoli seeds germinated at a different time and technological processing conditions that keep the activity of the enzyme to form sulforaphane. Broccoli seeds were germinated in the house. Myrosinase activities were tested as the glucose content using glucose assay kit and measured UV-Vis spectrophotometer. Glucosinolates were measured by HPLC/DAD. Sulforaphane was measured using HPLC-DAD and GC/MS. The 6 hr germinated sprouts have a myrosinase activity 32.2 mg glucose/g, which is comparable with 12 and 24 hour germinated seeds and higher than dry seeds. The glucoraphanin content in 6 hour germinated sprouts is 13935 µg/g which is comparable to 24 hour germinated seeds and lower than the dry seeds. GC/MS results show that the amount of sulforaphane is higher than the amount of sulforaphane nitrile in seeds, 6 hour and 24 hour germinated seeds. The ratio of sulforaphane and sulforaphane nitrile is high in 6 hour germinated seeds, which indicates the inactivated ESP in the reaction. After evaluating the results, the short time germinated seeds can be used as the source of glucoraphanin and myrosinase supply to form potential higher sulforaphane content. Broccoli contains glucosinolates, glucoraphanin (4-methylsulfinylbutyl glucosinolate), which is an important metabolite with health-promoting effects. In the pilot clinical study, we observed the effects of a glucosinolates/glucoraphanin-rich extract from short time germinated broccoli seeds on blood adenosine triphosphate (ATP), reactive oxygen species (ROS) and lactate levels. A single dose of 50 mg of broccoli sprouts extract increased blood levels of ATP up to 61% (p=0.0092) during the first 2 hours after the ingestion. Interestingly, this effect was not associated with an increase in blood ROS or lactate. When compared to the placebo group, levels of lactate were reduced by 10% (p=0.006). These results indicate that broccoli germinated seed extract may positively affect the generation of ATP in humans. Due to the preliminary nature of this work and promising results, larger clinical trials are justified.

Keywords: broccoli glucosinolates, glucoraphanin, germinated seeds, myrosinase, adenosine triphosphate

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Authors: Yashvir Singh, Amneesh Singla, Swapnil Bhurat

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Around the globe, there is demand for the development of bio-based lubricant which will be biodegradable, non toxic, and environmentally-friendly. This paper outlines the friction and wear characteristics of ponagamia biodiesel contaminated bio-lubricant by using pin-on-disc tribometer. To formulate the bio-lubricants, Ponagamia oil based biodiesel were blended in the ratios 5, 10, and 20% by volume with the base lubricant SAE 20 W 40. Tribological characteristics of these blends were carried out at 2.5 m/s sliding velocity and loads applied were 50, 100, 150 N. Experimental results showed that the lubrication regime that occurred during the test was boundary lubrication while the main wear mechanisms was the adhesive wear. During testing, the lowest wear was found with the addition of 5 and 10% Ponagamia oil based biodiesel, and above this contamination, the wear rate was increased considerably. The addition of 5 and 10% Ponagamia oil based biodiesel with the base lubricant acted as a very good lubricant additive which reduced the friction and wear rate during the test. It has been concluded that the PBO 5 and PBO 10 can act as an alternative lubricant to increase the mechanical efficiency at 2.5 m/s sliding velocity and contribute in reduction of dependence on the petroleum based products.

Keywords: friction, load, pongamia oil blend, sliding velocity, wear

Procedia PDF Downloads 296

Authors: Fariba Rahimi, Morteza Zendedel, Mohammad Jaafar Rezaee, Bita Vazir, Shahin Fakour

Abstract:

The present study was carried out to investigate the effect of melatonin (Mel) with and without zinc (Zn) on the gonadotropin hormones, also thyroid (T3 and T4) hormone concentration in female rats. A total of 40 adult female rats were randomly grouped into five treatment groups, each of 2 rats in a Completely Randomized Design (CRD) entire research time. Daily was treated by gavage with Zn and melatonin as follows: T1 (control1, basal diet), T2 (control 2, treated with normal saline) and other experimental groups, including T3, T4 and T5, were treated with a dose of zinc (40 ppm), melatonin (5 mg/kg), and combination zinc plus melatonin with the same level, respectively. Blood FSH and LH concentrations were measured. The result showed no significant differences between treatments in FSH and LH levels. The estrogen and progesterone and TSH levels in rats that received 5 mg of melatonin per day were higher than in other groups but not statistically significant (P>0.05). However, T3 (thyroid) concentration significantly (P<0.05) decreased in the group that received 40 mg/zinc per Kg compared to other groups. No significant (P>0.05) difference was detected among treatments in T4 levels. In conclusion, except for T3, had no significant (P>0.05) effect on another parameter in the female rats that received melatonin or zinc and a blend of melatonin and Zn.

Keywords: zinc, melatonin, hormone, rat

Procedia PDF Downloads 83

Authors: Pinar Ozdemir Ayber, Zeina Hojeij

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Blended Learning (BL) is a rapidly growing teaching and learning approach, which brings together the best of both face-to-face and online learning to expand learning opportunities for students. However, there is limited research on the practices, opportunities and quality of instruction in Blended Classrooms, and on the role of the teaching faculty as well as the learners in these types of classes. This paper will highlight the researchers’ experiences and reflections on blending their classes. It will focus on the importance of designing effective lesson plans that emphasize learner engagement and motivation in alignment with course learning outcomes. In addition, it will identify the changing roles of the teacher and the learners and suggest appropriate variations to the traditional classroom setting taking into consideration the benefits and the challenges of the Blended Classroom. It is hoped that this paper would provide sufficient input for participants to reflect on ways they can blend their own lessons to promote ubiquitous learning and student autonomy. Practical tips and ideas will be shared with the participants on various strategies and technologies that were used in the researchers’ classes.

Keywords: blended learning, learner autonomy, learner engagement, learner motivation, mobile learning tools

Procedia PDF Downloads 279

Authors: Pao-Huei Chen, Shu-Mei Lin, Yih-Ming Weng, Zer-Ran Yu, Be-Jen Wang

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Pancreatic α-amylase and intestinal α-glucosidase are the critical enzymes for the breakdown of complex carbohydrates into di- or mono-saccharide, which play an important role in modulating postprandial blood sugars. Angiotensin converting enzyme (ACE) converts inactive angiotensin-I into active angiotensin-II, which subsequently increase blood pressure through triggering vasoconstriction and aldosterone secretion. Thus, inhibition of carbohydrate-digestion enzymes and ACE will help the management of blood glucose and blood pressure, respectively. Studies showed Pleurotus citrinopileatus (PC), an edible mushroom and commonly cultured in oriental countries, exerted anticancer, immune improving, antioxidative, hypoglycemic and hypolipidemic effects. Previous studies also showed various molecular weights (MW) fractioned from extracts may affect biological activities due to varying contents of bioactive components. Thus, the objective of this study is to investigate the in vitro antioxidant, hypoglycemic and hypotenstive effects and distribution of active compounds of various MWs of cold water extract from P. citrinopileatus (CWEPC). CWEPC was fractioned into four various MW fractions, PC-I (＜1 kDa), PC-II (1-3.5 kDa), PC-III (3.5-10 kDa), and PC-IV (＞10 kDa), using an ultrafiltration system. The physiological activities, including antioxidant activities, the inhibition capabilities of pancreatic α-amylase, intestinal α-glucosidase, and hypertension-linked ACE, and the active components, including polysaccharides, protein, and phenolic contents, of CWEPC and four fractions were determined. The results showed that fractions with lower MW exerted a higher antioxidant activity (p<0.05), which was positively correlated to the levels of total phenols. In contrast, the inhibition effects on the activities of α-amylase, α-glucosidase, and ACE of PC-IV fraction were significantly higher than CWEPC and the other three low MW fractions (< 10 kDa), which was more related to protein contents. The inhibition capability of CWEPC and PC-IV on α-amylase activity was 1/13.4 to 1/2.7 relative to that of acarbose (positive control), respectively. However, the inhibitory ability of PC-IV on α-glucosidase (IC50 = 0.5 mg/mL) was significantly higher than acarbose (IC50 = 1.7 mg/mL). Kinetic data revealed that PC-IV fraction followed a non-competitive inhibition on α-glucosidase activity. In conclusion, the distribution of various bioactive components contribute to the functions of different MW fractions on oxidative stress prevention, and blood pressure and glucose modulation.

Keywords: α-Amylase, angiotensin converting enzyme, α-Glucosidase, Pleurotus citrinopileatus

Procedia PDF Downloads 445

Authors: Flordeliza L. Mercado, Teresito G. Aguinaldo, Helen F. Gavino, Victorino T. Taylan

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Most fruits and vegetables are available in large quantities during peak season which are oftentimes marketed at low price and left to rot or fed to farm animals. The lack of efficient storage facilities, and the additional cost and unavailability of small machinery for food processing, results to low price and wastage. Incidentally, processed fresh fruits and vegetables are gaining importance nowadays and health conscious people are also into ‘juicing’. One way to reduce wastage and ensure an all-season availability of crop juices at reasonable costs is to develop equipment for effective extraction of juice. The study was conducted to design, fabricate and evaluate a multi-juice extractor using locally available materials, making it relatively cheaper and affordable for medium-scale enterprises. The study was also conducted to formulate juice blends using extracted juices and calamansi juice at different blending percentage, and evaluate its chemical properties and sensory attributes. Furthermore, the chemical properties of extracted meals were evaluated for future applications. The multi-juice extractor has an overall dimension of 963mm x 300mm x 995mm, a gross weight of 82kg and 5 major components namely; feeding hopper, extracting chamber, juice and meal outlet, transmission assembly, and frame. The machine performance was evaluated based on juice recovery, extraction efficiency, extraction rate, extraction recovery, and extraction loss considering type of crop as apple and carrot with three replications each and was analyzed using T-test. The formulated juice blends were subjected to sensory evaluation and data gathered were analyzed using Analysis of Variance appropriate for Complete Randomized Design. Results showed that the machine’s juice recovery (73.39%), extraction rate (16.40li/hr), and extraction efficiency (88.11%) for apple were significantly higher than for carrot while extraction recovery (99.88%) was higher for apple than for carrot. Extraction loss (0.12%) was lower for apple than for carrot, but was not significantly affected by crop. Based on adding percentage mark-up on extraction cost (Php 2.75/kg), the breakeven weight and payback period for a 35% mark-up is 4,710.69kg and 1.22 years, respectively and for a 50% mark-up, the breakeven weight is 3,492.41kg and the payback period is 0.86 year (10.32 months). Results on the sensory evaluation of juice blends showed that the type of juice significantly influenced all the sensory parameters while the blending percentage including their respective interaction, had no significant effect on all sensory parameters, making the apple-calamansi juice blend more preferred than the carrot-calamansi juice blend in terms of all the sensory parameter. The machine’s performance is higher for apple than for carrot and the cost analysis on the use of the machine revealed that it is financially viable with a payback period of 1.22 years (35% mark-up) and 0.86 year (50% mark-up) for machine cost, generating an income of Php 23,961.60 and Php 34,444.80 per year using 35% and 50% mark-up, respectively. The juice blends were of good qualities based on the values obtained in the chemical analysis and the extracted meal could also be used to produce another product based on the values obtained from proximate analysis.

Keywords: food processing, fruits and vegetables, juice extraction, multi-juice extractor

Procedia PDF Downloads 279

Authors: Chandrashekara Krishnappa, Yogish Huchaiah

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The present work considers the importance of economic feasibility and financial viability of biodiesel production, and its use in the present context of prevailing Indian scenario. For this, costs involved in production of one litre of biodiesel from non-edible Jatropha and Pongamia oils Nano mix are considered. Biodiesel derived from the mix is blended with petrodiesel in various proportions and used in Compression Ignition (CI) Direct Injection (DI) engine. Performance and Emission characteristics were investigated. Optimization of the blends considering experimental results was carried out. To validate the experimental results and optimization, Multi-Functional Criteria Technique (MFCT) is used. Further, value additions in terms of INR due to increase in performance and reduction in emissions are investigated. Cost component of subsidy on petrodiesel is taken into consideration in the calculation of cost of one litre of it. Comparison of costs is with respect to the unit of power generated per litre of COME and petrodiesel. By the analysis it has been concluded that the amount saved with subsidy is INR 1.45 Lakh Crores per year and it is INR1.60 Lakh Crores per year without subsidy for petrodiesel.

Keywords: cap value addition, economic analysis, MFCT, NACOME, subsidy

Procedia PDF Downloads 223

Authors: Hamzeh Alipour, Masoumeh Bagheri, Abbasali Raz, Javad Dadgar Pakdel, Kourosh Azizi, Aboozar Soltani, Mohammad Djaefar Moemenbellah-Fard

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Background: Maggot debridement therapy is an appropriate, effective, and controlled method using sterilized larvae of Luciliasericata (L.sericata) to treat wounds. Netrin-A is an enzyme in the Laminins family which secreted from salivary gland of L.sericata with a central role in neural regeneration and angiogenesis. This study aimed to production of new recombinant Netrin-A protein of Luciliasericata larvae by baculovirus expression vector system (BEVS) in SF9. Material and methods: In the first step, gene structure was subjected to the in silico studies, which were include determination of Antibacterial activity, Prion formation risk, homology modeling, Molecular docking analysis, and Optimization of recombinant protein. In the second step, the Netrin-A gene was cloned and amplified in pTG19 vector. After digestion with BamH1 and EcoR1 restriction enzymes, it was cloned in pFastBac HTA vector. It was then transformed into DH10Bac competent cells, and the recombinant Bacmid was subsequently transfected into insect Sf9 cells. The expressed recombinant Netrin-A was thus purified in the Ni-NTA agarose. This protein evaluation was done using SDS-PAGE and western blot, respectively. Finally, its concentration was calculated with the Bradford assay method. Results: The Bacmid vector structure with Netrin-A was successfully constructed and then expressed as Netrin-A protein in the Sf9 cell lane. The molecular weight of this protein was 52 kDa with 404 amino acids. In the in silico studies, fortunately, we predicted that recombinant LSNetrin-A have Antibacterial activity and without any prion formation risk.This molecule hasa high binding affinity to the Neogenin and a lower affinity to the DCC-specific receptors. Signal peptide located between amino acids 24 and 25. The concentration of Netrin-A recombinant protein was calculated to be 48.8 μg/ml. it was confirmed that the characterized gene in our previous study codes L. sericata Netrin-A enzyme. Conclusions: Successful generation of the recombinant Netrin-A, a secreted protein in L.sericata salivary glands, and because Luciliasericata larvae are used in larval therapy. Therefore, the findings of the present study could be useful to researchers in future studies on wound healing.

Keywords: blowfly, BEVS, gene, immature insect, recombinant protein, Sf9

Procedia PDF Downloads 70

Authors: J. O. Idoko, I. Nwajiaku

Abstract:

Unripe and ripe plantain were dried and milled into flour and used with wheat flour in biscuit production to determine the best plantain-wheat composite flour for biscuit production. The blends as follows: 100% wheat flour, 100% ripe plantain flour, 100% unripe plantain flour, 50% wheat flour and 50% ripe plantain flour and 50% wheat flour and 50% unripe plantain flour. The Biscuit samples were stored at ambient temperature for 8 weeks after which the equilibrium moisture content and water activity were determined. The sensory evaluation of the biscuit samples was also determined. The results of these analyses showed 100% unripe plantain flour as the most stable of the biscuit samples judging from its equilibrium moisture content level of 0.32% and water activity of 0.62. The sensory evaluation results showed Biscuit made from 150:50 ripe plantain and wheat flour as most generally accepted at 5% level of significance.

Keywords: biscuit, equilibrium moisture content, performance, plantain, water activity

Procedia PDF Downloads 183

Authors: Fariba Rahimi, Morteza Zendedel, Mohammad Jaafar Rezaee, Bita Vazir, Shahin Fakour

Abstract:

The present study was carried out to investigate the effect of melatonin (Mel) with and without zinc (Zn) on the gonadotropin hormones, also thyroid (T3 and T4) hormone concentration in female rat. A total of 40 adult female rats were randomly grouped into five treatment groups, each of 2 rats in a Completely Randomized Design (CRD) entire research time. Daily were treated by gavaging with Zn and melatonin as following: T1 (control1, basal diet), T2 (control 2, treated with normal saline) and other experimental groups including T3, T4 and T5 were treated with dose of zinc (40 ppm), melatonin (5 mg/kg), and combination zinc plus melatonin with the same level, respectively. Blood FSH and LH concentration were measured. Result showed no significantly differences between treatments in FSH and LH levels. The estrogen and progesterone and TSH levels in rats that received 5 mg of melatonin per day were higher than other groups, but not statistically significant (P>0.05). However, T3 (thyroid) concentration significantly (P<0.05) decreased in group that received 40 mg/zinc per Kg compared other groups. No significant (P>0.05) difference was detected among treatments in T4 levels. In conclusion, except for T3, had not significantly (P>0.05) effect on another parameters in the female rats that received melatonin or zinc and blend of melatonin and Zn.

Keywords: zinc, melatonin, hormone, rat

Procedia PDF Downloads 93