Search results for: enzyme pre-treatment

Authors: Celestine Mbakaan, Iorkyaa Ahemen, A. D. Onoja, A. N. Amah, Emmanuel Barki

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Rice husk (RH) is a natural sheath that forms and covers the grain of rice. The husk composed of hard materials, including opaline silica and lignin. It separates from its grain during rice milling. RH also contains approximately 15 to 28 wt % of silica in hydrated amorphous form. Nanosilica was derived from the husk of different rice varieties after pre-treating the husk (RH) with HCl and calcination at 550°C. Nanosilica derived from the husk of Osi rice variety produced the highest silica yield, and further pretreatment with 0.8 M H₃PO₄ acid removed more mineral impurities. The silica obtained from this rice variety was selected as a host matrix for doping with Sm³⁺ ions. Rice husk silica (RH-SiO₂) doped with samarium (RH-SiO₂: xSm³⁺ (x=0.01, 0.05, and 0.1 molar ratios) nanophosphors were synthesized via the sol-gel method. The structural analysis by X-ray diffraction analysis (XRD) reveals amorphous structure while the surface morphology, as revealed by SEM and TEM, indicates agglomerates of nano-sized spherical particles with an average particle size measuring 21 nm. The nanophosphor has a large surface area measuring 198.0 m²/g, and Fourier transform infrared spectroscopy (FT-IR) shows only a single absorption band which is strong and broad with a valley at 1063 cm⁻¹. Diffuse reflectance spectroscopy (DRS) shows strong absorptions at 319, 345, 362, 375, 401, and 474 nm, which can be exclusively assigned to the 6H5/2→4F11/2, 3H7/2, 4F9/2, 4D5/2, 4K11/2, and 4M15/2 + 4I11/2, transitions of Sm³⁺ respectively. The photoluminescence excitation spectra show that near UV and blue LEDs can effectively be used as excitation sources to produce red-orange and yellow-orange emission from Sm³⁺ ion-doped RH-SiO₂ nanophosphors. The photoluminescence (PL) of the nanophosphors gives three main lines; 568, 605, and 652 nm, which are attributed to the intra-4f shell transitions from the excited level to ground levels, respectively under excitation wavelengths of 365 and 400 nm. The result, as confirmed from the 1931 CIE coordinates diagram, indicates the emission of red-orange light by RH-SiO₂: xSm³⁺ (x=0.01 and 0.1 molar ratios) and yellow-orange light from RH-SiO₂: 0.05 Sm³⁺. Finally, the result shows that RH-SiO₂ doped with samarium (Sm³⁺) ions can be applicable in display applications.

Keywords: luminescence, nanosilica, nanophosphors, Sm³⁺

Procedia PDF Downloads 121

Authors: P. O. Ughachukwu, P. O. Okonkwo, P. C. Unekwe, J. O. Ogamba

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Background: The prevalence of hepatitis B viral infection is high worldwide with liver cirrhosis and hepatocellular carcinoma as important complications. Cases of poor antibody response to hepatitis B vaccination abound. Immunosuppression, especially from glucocorticoids, is often cited as a cause of poor antibody response and there are documented evidences of irrational administration of glucocorticoids to children and adults. The study was, therefore, designed to find out if administration of glucocorticoids affects immune response to vaccination against hepatitis B in mice. Methods: Mice of both sexes were randomly divided into 2 groups. Daily intramuscular methylprednisolone injections, (15 mg kg-1), were given to the test group while sterile deionized water (0.1ml) was given to control mice for 30 days. On day 6 all mice were given 2 μg (0.1ml) hepatitis B vaccine and a booster dose on day 27. On day 34, blood samples were collected and analyzed for anti-HBs titres using enzyme-linked immunosorbent assay (ELISA). Statistical analysis was done using Graph Pad Prism 5.0 and the results taken as statistically significant at p value < 0.05. Results: There were positive serum anti-HBs responses in all mice groups but the differences in titres were not statistically significant. Conclusions: At the dosages and length of exposure used in this study, methylprednisolone injection did not significantly inhibit anti-HBs response in mice following immunization against hepatitis B virus. By extrapolation, methylprednisolone, when used in the usual clinical doses and duration of therapy, is not likely to inhibit immune response to hepatitis B vaccinations in man.

Keywords: anti-HBs, hepatitis B vaccine, immune response, methylprednisolone, mice

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Authors: Mirnawati, G. Ciptaan, Ferawati

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This study aims to improve the quality of the cassava peel-leaf mixture (CPLM) through fermentation with Rhizopus oligosporusas poultry ration. This research is an experimental study using a completely randomized design (CRD) with four treatments and five replications. The treatments were cassava peel-leaf mixture (CPLM) fermented with Rhizopus oligosporus. The treatments were a combination of cassava peel and leaves with the ratio of; A (9:1), B (8:2), C (7:3), and D (6:4). The observed variables were protease enzyme activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and metabolic energy. The results of the diversity analysis showed that there was a very significant (p < 0.01) effect on protease activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and energy metabolism of fermented CPLM. Based on the results of the study, it can be concluded that CPLM (6:4) fermented with Rhizopus oligosporus gave the best results seen from protease activity 7,25 U/ml, 21.23% crude protein, 19.80% crude fiber, 59.65% nitrogen retention, 62.99% crude fiber digestibility and metabolic energy 2671 Kcal/kg.

Keywords: quality, Casava peel-leaf mixture, fermentation, Rhizopus oligosporus

Procedia PDF Downloads 165

Authors: S. Hallal, Z. Chami, A. Hadji-Lehtihet, S. Sokhal-Boudella, A. Berhoune, L. Yargui

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Gaucher disease is the most common lysosomal storage in our population, it is due to a deficiency of β –glucosidase acid. The enzyme deficiency causes a pathological accumulation of undegraded substrate in lysosomes. This metabolic overload is responsible for a multisystemic disease with hepatosplenomegaly, anemia, thrombocytopenia, and bone involvement. Neurological involvement is rare. The laboratory diagnosis of Gaucher disease consists of phenotypic diagnosis by determining the enzymatic activity of β - glucosidase by fluorimetric method, a study by genotypic diagnosis in the GBA gene, limiting the search recurrent mutations (N370S, L444P, 84 GG); PCR followed by an enzymatic digestion. Abnormal profiles were verified by sequencing. Monitoring of treated patients is provided by the determination of chitotriosidase. Our experience spaning a period of 6 years (2007-2014) has enabled us to diagnose 78 patients out of a total of 328 requests from the various departments of pediatrics, internal medicine, neurology. Genotypic diagnosis focused on the entire family of 9 children treated at pediatric CHU Mustapha, which help define the clinical form; or 5 of them had type III disease, carrying the L444P mutation in the homozygous state. Three others were composite (N370/L444P) (N370S/other unintended mutation in our study), and only in one family no recurrent mutation has been found. This molecular study permits screening of heterozygous essential for genetic counseling.

Keywords: Gaucher disease, mutations, N370S, L444P

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Authors: Naso Isaiah Thanavisuth

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Medical cannabis can be used for treatment including pain, multiple sclerosis, Parkinson's disease, and cancer. However, medical cannabis leads to adverse effects (AEs), which is delta-9-tetrahydrocannabinol (THC). In previous studies, the major of THC metabolism enzymes are CYP2C9. Especially, the variation of CYP2C9 gene consist of CYP2C9*2 on exon 3 and CYP2C9*3 on exon 7 to decrease enzyme activity. Notwithstanding, there is no data describing whether the variant of CYP2C9 genes are apharmacogenetics marker for the prediction of THC-induced AEs in Thai patients. We want to investigate the association between CYP2C9 gene and THC-induced AEs in Thai patients. We enrolled 39 Thai patients with medical cannabis treatment who were classified by clinical data. The CYP2C9*2 and *3 genotyping were conducted using the TaqMan real time PCR assay. All Thai patients who received the medical cannabis consist of twenty-four (61.54%) patients were female, and fifteen (38.46%) were male, with age range 27- 87 years. Moreover, the most AEs in Thai patients who were treated with medical cannabis between cases and controls were tachycardia, arrhythmia, dry mouth, and nausea. Particularly, thirteen (72.22%) medical cannabis-induced AEs were female and age range 33 – 69 years. In this study, none of the medical cannabis groups carried CYP2C9*2 variants in Thai patients. The CYP2C9*3 variants (*1/*3, intermediate metabolizer, IM) and (*3/*3, poor metabolizer, PM) were found, three of thirty-nine (7.69%) and one of thirty-nine (2.56%), respectively. Although, our results indicate that there is no found the CYP2C9*2. However, the variation of CYP2C9 allele might serve as a pharmacogenetics marker for screening before initiating the therapy with medical cannabis for the prevention of medical cannabis-induced AEs.

Keywords: CYP2C9, medical cannabis, adverse effects, THC, P450

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Authors: Hawon Lee, Young-Pil Kim

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Visualizing matrix metalloproteinases (MMPs) activity is necessary for understanding cancer metastasis because they are implicated in cell migration and invasion by degrading the extracellular matrix (ECM). While much effort has been made to sense the MMP activity, but extracellularly long-term monitoring of MMP activity still remains challenging. Here, we report a collagen-bound fluorescent bioprobe for the detection of MMP-2 activity in the extracellular environment. This bioprobe consists of ECM-immobilized part (including collagen-bound protein) and MMP-sensing part (including peptide substrate linked with fluorescence resonance energy transfer (FRET) coupler between donor green fluorescent protein (GFP) and acceptor TAMRA dye), which was constructed through intein-mediated self-splicing conjugation. Upon being immobilized on the collagen-coated surface, this bioprobe enabled efficient long-lasting observation of MMP-2 activity in the cultured cells without affecting cell growth and viability. As a result, the FRET ratio (acceptor/donor) decreased as the MMP2 activity increased in cultured cancer cells. Furthermore, unlike wild-type MMP-2, mutated MMP-2 expression (Y580A in the hemopexin region) gave rise to lowering the secretion of MMP-2 in HeLa. Conclusively, our method is anticipated to find applications for tracing and visualizing enzyme activity.

Keywords: collagen, ECM, FRET, MMP

Procedia PDF Downloads 187

Authors: Namsoo Kim, So-Hee Son, Jin-Soo Maeng, Yong-Jin Cho, Chong-Tai Kim

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It has been reported that some enzymes such as trypsin and Alcalase 2.4L are tolerant to a medium high pressure of 300 MPa and preparation of protein hydrolyzates under 300 MPa was advantageous with regard to hydrolysis rate and thus production yield compared with the counterpart under ambient pressure.1,2) In this study, nine fish comprising halibut, soft shell clam and carp were hydrolyzed using Flavourzyme 500MG only, and the combination of Flavourzyme 500 mg, Alcalase 2.4 L, Marugoto E, and Protamex under 300 MPa. Then, the effects of single and multiple protease treatments were determined with respect to contents of soluble solid (SS) and soluble nitrogen, sensory attributes, electrophoretic profiles, and HPLC peak patterns of the fish hydrolyzates (FHs) from various species. The contents of SS of the FHs were quite species-specific and the hydrolyzates of halibut showed the highest SS contents. At this point, multiple protease treatment increased SS content conspicuously in all fish tested. The contents of total soluble nitrogen and TCA-soluble nitrogen were well correlated with those of SS irrespective of fish species and methods of enzyme treatment. Also, it was noticed that multiple protease treatment improved sensory attributes of the FHs considerably. Electropherograms of the FHs showed fast migrating peptide bands that had the molecular masses mostly lower than 1 kDa and this was confirmed by peptide patterns from HPLC analysis for some FHs that had good sensory quality.

Keywords: production, fish hydrolyzates, protease treatments, high pressure

Procedia PDF Downloads 265

Authors: Mei-Ying Huang, Huei-Jen Ju, Liang-Wei Tseng, Chin-Jung Hsu

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The aim of this study was to investigate a probiotic, Leuconostoc mesenteroides B4, and its products, isomaltooligosaccharide and dextran, on growth performance, digestive enzymes, immune responses, and pathogen resistance of spotted grouper Epinephelus coioides. The grouper were fed control and diets supplemented with L. mesenteroides B4 (107 CFU/g), isomaltooligosaccharide (0.15%), isomaltooligosaccharide (0.15%) + L. mesenteroides B4 (107 CFU/g) (I + B4), and dextran (0.15%) + L. mesenteroides B4 (107 CFU/g) (D + B4) for 8 weeks. The result showed that final weights and percent weight gains of the grouper fed diets supplemented with L. mesenteroides B4 and I + B4 were significantly higher than that of the control group (p < 0.05). The activities of digestive enzymes in the grouper fed with I + B4 were significantly higher than the control group (p < 0.05), too. After challenge with Vibrio harveyi, the enzyme activities of antiprotease and lysozyme as well as of respiratory burst of the fish fed with I + B4 and D + B4 were significantly higher than that of the control group (p < 0.05). The grouper fed with the both diets also had higher survival rates than that of the control group after the challenge. Overall, the study indicated that feeding diets supplemented with L. mesenteroides B4, and its products, isomaltooligosaccharide, and dextran could be an effective method for enhancing the growth performance and disease resistance in orange-spotted grouper.

Keywords: orange-spotted grouper, probiotic Leuconostoc mesenteroides B4, isomaltooligosaccharide, dextran, growth performance, pathogen resistance

Procedia PDF Downloads 249

Authors: Atena Naeimi, Mehri-Sadat Ekrami-Kakhki

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Fe3O4 is one of the important magnetic oxides with spinel structure; it has exhibited unique electric and magnetic properties based on the electron transfer between Fe2+ and Fe3+ in the octahedral sites. Fe3O4 have received considerable attention in various areas such as cancer therapy, drug targeting, enzyme immobilization catalysis, magnetic cell separation, magnetic refrigeration systems and super-paramagnetic materials. Fe3O4–ZnO nanostructures were synthesized via a surfactant-free ultrasonic reaction at room temperatures. The effect of various parameters such as temperature, time, and power on the size and morphology of the product was investigated. Alternating gradient force magnetometer shows that Fe3O4 nanoparticles exhibit super-paramagnetic behaviour at room temperature. For preparation of nanocomposite 1 g of Fe3O4 nanostructures were dispersed in 100 mL of distilled water. 0.25 g of Zn (NO3)2 and 20 mL of NH3 solution 1 M were then slowly added to the solution under ultrasonic irradiation. The product was centrifuged, washed with distilled water and dried in the air. The photocatalytic behaviour of Fe3O4–ZnO nanoparticles was evaluated using the degradation of a methyl orange aqueous solution under ultraviolet light irradiation. As time increased, more and more methyl orange was adsorbed on the nanoparticles catalyst, until the absorption peak vanish. The methyl orange concentration decreased rapidly with increasing UV-irradiation time.

Keywords: nanocomposite, ultrasonic, paramagnetic, photocatalytic

Procedia PDF Downloads 289

Authors: Stella O. Olubodun, George E. Eriyamremu

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The study examined the effect of Bonny Light whole crude oil (WC) and its water soluble fraction (WSF) on the activities of antioxidant enzymes (catalase (CAT) and superoxide dismutase (SOD)) and crude mitochondria ATPases in the radicle of germinating bean (Vigna unguiculata). The percentage germination, level of lipid peroxidation, antioxidant enzyme, and mitochondria Ca2+ and Mg2+ ATPase activities were measured in the radicle of bean after 7, 14, and 21 days post germination. Viable bean seeds were planted in soils contaminated with 10ml, 25ml, and 50ml of whole crude oil (WC) and its water soluble fraction (WSF) to obtain 2, 5, and 10% v/w crude oil contamination. There was dose dependent reduction of the number of bean seeds that germinated in the contaminated soils compared with control (p<0.001). The activities of the antioxidant enzymes, as well as, adenosine triphosphatase enzymes, were also significantly (p<0.001) altered in the radicle of the plants grown in contaminated soil compared with the control. Generally, the level of lipid peroxidation was highest after 21 days post germination when compared with control. Stress to germinating bean caused by Bonny Light crude oil or its water soluble fraction resulted in adaptive changes in crude mitochondria ATPases in the radicle.

Keywords: antioxidant enzymes, bonny light crude oil, radicle, mitochondria ATPases

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Authors: Kshitij Bhardwaj, R.K. Trivedi, Shipra Dixit

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We used biological detoxification method that converts toxic residue waste of Jatropha curcas oil seeds (non edible oil seed) into industrial bio-products and animal feed material. Present study describes the complete degradation of phorbol esters by Aspergillus Niger strain during solid state fermentation (SSF) of deoiled Jatropha curcas seed cake. Phorbol esters were completely degraded in 15 days under the optimized SSF conditions viz deoiled cake 5.0 gm moistened with 5.0 ml distilled water; inoculum 2 ml of overnight grown Aspergillus niger; incubated at 30◦ C, pH 7.0. This method simultaneously induces the production of Protease enzyme by Aspergillus Niger which has high potential to be used in feedstuffs .The maximum Protease activities obtained were 709.16 mg/ml in Jatropha curcas oil seed cake. The protein isolate had small amounts of phorbol esters, phytic acid, and saponin without any lectin. Its minimum and maximum solubility were at pH 4.0&12.0. Water and oil binding capacities were 3.22 g water/g protein and 1.86 ml oil/g protein respectively.Emulsion activity showed high values in a range of basic pH. We concluded that Jatropha Curcas seed cake has a potential to be used as a novel source of functional protein for food or feed applications.

Keywords: solid state fermentation, Jatropha curcas, oil seed cake, phorbol ester

Procedia PDF Downloads 465

Authors: Akwu N. A., Naidoo Y., Salau V. F., Olofinsan K. A.

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Grewia lasiocarpa E. Mey. ex Harv. (Malvaceae) is a Southern African medicinal plant indigenously used with other plants for birthing problems. The anti-diabetic properties of the hexane, chloroform, and methanol extracts of Grewia lasiocarpa stem bark were assessed using in vitro α-glucosidase enzyme inhibition assay. The predictive in silico drug-likeness and toxicity properties of the phytocompounds were conducted using the pKCSM, ADMElab, and SwissADME computer-aided online tools. The highest α-glucosidase percentage inhibition was observed in the hexane extract (86.76%, IC50= 0.24 mg/mL), followed by chloroform (63.08%, IC50= 4.87 mg/mL) and methanol (53.22%, IC50= 9.41 mg/mL); while acarbose, the standard anti-diabetic drug was (84.54%, IC50= 1.96 mg/mL). The α-glucosidase assay revealed that the hexane extract exhibited the strongest carbohydrate inhibiting capacity and is a better inhibitor than the standard reference drug-acarbose. The computational studies also affirm the results observed in the in vitroα-glucosidaseassay. Thus, the extracts of G. lasiocarpa may be considered a potential plant-sourced compound for treating type 2 diabetes mellitus. This is the first study on the anti-diabetic properties of Grewia lasiocarpa hexane, chloroform, and methanol extracts using in vitro and in silico models.

Keywords: grewia lasiocarpa, α-glucosidase inhibition, anti-diabetes, ADMET

Procedia PDF Downloads 86

Authors: Hari Bezwada, Michelle Cheon, Ryan Divan, Hannah Escritor, Michelle Kagramian, Isha Korgaonkar, Maya MacAdams, Udgita Pamidigantam, Richard Pilny, Eleanor Race, Angadh Singh, Nathan Zhang, LeeAnn Nguyen, Fina Liotta

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Malaria is a deadly disease caused by the Plasmodium parasite, which continues to develop resistance to current antimalarial drugs. In this research project, the effectiveness of numerous thiazole derivatives was explored in inhibiting the PfPKG, a crucial part of the Plasmodium life cycle. This study involved the synthesis of six thiazole-derived amides to inhibit the PfPKG pathway. Nuclear Magnetic Resonance (NMR) spectroscopy and Infrared (IR) spectroscopy were used to characterize these compounds. Furthermore, AutoDocking software was used to predict binding affinities of these thiazole-derived amides in silico. In silico, compound 6 exhibited the highest predicted binding affinity to PfPKG, while compound 5 had the lowest affinity. Compounds 1-4 displayed varying degrees of predicted binding affinity. In-vitro, it was found that compound 4 had the best percent inhibition, while compound 5 had the worst percent inhibition. Overall, all six compounds had weak inhibition (approximately 30-39% at 10 μM), but these results provide a foundation for future drug discovery experiments.

Keywords: Medicinal Chemistry, Malaria, drug discovery, PfPKG, Thiazole, Plasmodium

Procedia PDF Downloads 71

Authors: Kriti Upadhyay, Ashraf Ali, Puja Sohal, Randeep Guleria

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Background: In Chronic Obstructive Pulmonary diseases (COPD) pathogenesis oxidative stress plays an important role. Hypoxia-Inducible factor (HIF-1α) is a dimeric protein complex which Functions as a master transcriptional regulator of the adaptive response to hypoxiaand is a risk factor that increases when oxidative stress triggers. The role ofHIF-1αin COPD due to smoking is lacking. Aim: This study aims to evaluate the role of HIF-1α in smoker COPD patients comparing its association with diseases severity. Method: In this cross-sectional study, we recruited 87 subjects, 57 were smokers with COPD,15 were smokers without COPD and other 15 were non-smoker healthy controls. The mean age was 54.6± 9.32 (cases 57.08±8.15; controls 50.0± 9.8). There were 62%smokers, 25% non-smokers,7% tobacco chewers and 6% ex-smokers. Enzyme-linked immune sorbent assay (ELISA) method was used for analyzing serum samples wherein HIF-1α was analyzed by Sandwich-ELISA. Results: In smoker COPD patients, a significantly higher HIF-1α level showed positive association with hypoxia, smoking status and severity of disease (p=0.03). The mean value of HIF-1α was not significantly different in smokers without COPD and healthy controls. Conclusion: It is found that HIF-1α level was increased in smoker COPD, but not in smokers without COPD. This suggests that development of COPD drive the HIF-1α pathway and it correlates with the severity of diseases.

Keywords: COPD, chronic obstructive pulmonary diseases, smokers, nonsmokers, hypoxia

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Authors: Chia-Jung Li, Kuan-Hao Tsui

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As women age, the quality of their oocytes deteriorates irreversibly, leading to reduced fertility. To better understand the role of Ferroptosis-related genes in ovarian aging, we employed a multi-omics analysis approach, including spatial transcriptomics, single-cell RNA sequencing, human ovarian pathology, and clinical biopsies. Our study identified excess lipid peroxide accumulation in aging germ cells, metal ion accumulation via oxidative reduction, and the interaction between ferroptosis and cellular energy metabolism. We used multi-histological prediction of ferroptosis key genes to evaluate 75 patients with ovarian aging insufficiency and then analyzed changes in hub genes after supplementing with DHEA, Ubiquinol CoQ10, and Cleo-20 T3 for two months. Our results demonstrated a significant increase in TFRC, GPX4, NCOA4, and SLC3A2, which were consistent with our multi-component prediction. We theorized that these supplements increase the mitochondrial tricarboxylic acid cycle (TCA) or electron transport chain (ETC), thereby increasing antioxidant enzyme GPX4 levels and reducing lipid peroxide accumulation and ferroptosis. Overall, our findings suggest that supplementation intervention significantly improves IVF outcomes in senescent cells by enhancing metal ion and energy metabolism and enhancing oocyte quality in aging women.

Keywords: multi-omics, nutrients, ferroptosis, ovarian aging

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Authors: Mandeep Kataria, Ritu Narula, Navneet Kaur

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Asparagine is vital amino acid which is required for the development of brain and it regulates the equilibrium of central nervous system. Asparagine is the chief amino acid that forms acrylamide in baked food by reacting with reducing sugars at high temperature ( Millard Reaction i.e. amino acids and sugars give new flavors at high temperature). It can also be a parameter of freshness in fruit juices because on storage of juices at 37°C caused an 87% loss in the total free amino acids and major decrease was recorded in asparagine contents. With this significance of monitoring asparagine, in the present work a biosensor for determining asparagine in fruit juices is developed. For the construction of biosensor L-asparaginase enzyme (0.5 IU) was co-immobilized with phenol red on TEOS chitosan sol-gel plastic disc and fixed on the fiber optic tip. Tip was immersed in a cell having 5ml of substrate and absorption was noted at response time of 5 min with 10-1 - 10-10 M concentrations of asparagine at 538 nm. L-asparaginase was extracted and from Solanum nigrum Asparagine biosensor was applied fruit juices on the monitoring asparagine contents. L-asparagine concentration found to be present in fruit juices like Guava Juice, Apple Juice, Mango Juice, Litchi juice, Strawberry juice, Pineapple juice Lemon juice, and Orange juice. Hence the developed biosensor has commercial aspects in quality insurance of fruit juices.

Keywords: fiber optic biosensor, chitosan, teos, l-asparaginase

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Authors: Jannes Quer, Amir Niknejad, Marcus Weber

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Computational Drug Design is often based on Molecular Dynamics simulations of molecular systems. Molecular Dynamics can be used to simulate, e.g., the binding and unbinding event of a small drug-like molecule with regard to the active site of an enzyme or a receptor. However, the time-scale of the overall binding event is many orders of magnitude longer than the time-scale of simulation. Thus, there is a need to speed-up molecular simulations. In order to speed up simulations, the molecular dynamics trajectories have to be ”steared” out of local minimizers of the potential energy surface – the so-called metastabilities – of the molecular system. Increasing the kinetic energy (temperature) is one possibility to accelerate simulated processes. However, with temperature the entropy of the molecular system increases, too. But this kind ”stearing” is not directed enough to stear the molecule out of the minimum toward the saddle point. In this article, we give a new mathematical idea, how a potential energy surface can be changed in such a way, that entropy is kept under control while the trajectories are still steared out of the metastabilities. In order to compute the unsteared transition behaviour based on a steared simulation, we propose to use extrapolation methods. In the end we mathematically show, that our method accelerates the simulations along the direction, in which the curvature of the potential energy surface changes the most, i.e., from local minimizers towards saddle points.

Keywords: extrapolation, Eyring-Kramers, metastability, multilevel sampling

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Authors: Serdal Pamuk, Irem Cay

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Our model is based on the theory of reinforced random walks coupled with Michealis-Menten mechanisms which view endothelial cell receptors as the catalysts for transforming both tumor and macrophage derived tumor angiogenesis factor (TAF) into proteolytic enzyme which in turn degrade the basal lamina. The model consists of two main parts. First part has seven differential equations (DE’s) in one space dimension over the capillary, whereas the second part has the same number of DE’s in two space dimensions in the extra cellular matrix (ECM). We connect these two parts via some boundary conditions to move the cells into the ECM in order to initiate capillary formation. But, when does this movement begin? To address this question we estimate the thresholds that activate the transport equations in the capillary. We do this by using steady-state analysis of TAF equation under some assumptions. Once these equations are activated endothelial, pericyte and macrophage cells begin to move into the ECM for the initiation of angiogenesis. We do believe that our results play an important role for the mechanisms of cell migration which are crucial for tumor angiogenesis. Furthermore, we estimate the long time tendency of these three cells, and find that they tend to the transition probability functions as time evolves. We provide our numerical solutions which are in good agreement with our theoretical results.

Keywords: angiogenesis, capillary formation, mathematical analysis, steady-state, transition probability function

Procedia PDF Downloads 144

Authors: Mansour F. Hussein, Mohammed A. Alshaikh, Riyadh S. Al-Jumaah, A. GarelNabi, I. Al-Khalifa, Osama B. Mohammed

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Serum samples from 489 male and female camels were tested for antibodies against C. burnetii using indirect enzyme-linked immunosorbent assay (ELISA). Antibodies to C. burnetii were recorded in sera of 252 (51.64%) camels. Significant differences in prevalence were found between male and female camels, juvenile and adult camels, different ecotypes and different sampling locations. 307 camels were simultaneously tested for C. burnetii antibodies by ELISA and indirect immunofluorescence (IFA). Close agreement was found between the results of the two tests. A high prevalence of C. burnetii antibodies was also recorded in milk samples tested by ELISA. Clinical samples from serologically positive camels were subjected to PCR analysis using primers which amplify the repetitive transposon-like and transposase gene regions of C. burnetii. Positive DNA amplification was obtained from both regions, with highest shedding of C. burnetii in faecal samples (27.59%) followed, in descending order, by urine (23.81%), blood (15.85%) and milk (6.5%). The present results indicate that camels are a major reservoir of C. burnetii in Saudi Arabia. The high prevalence of infection in camels, the poor sanitary standards under which the animals are kept and the consumption of raw camel milk indicate that camels could also be a major source of transmission of Q fever to humans in Saudi Arabia.

Keywords: Arabian camel, Camelus dromedarius, Coxiella brunetii, ELISA, immunofluoresence, PCR

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Authors: Afsoun Nikravan, Parisa Babaei, Gulen Gullu

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We aimed to study the association between house dust endotoxin, β-(1→3)-D-glucan, and asthma in a sample representative of the Turkish population. We analyzed data from 240 participants. The house dust was collected from the homes of 110 asthmatics and 130 control (without asthma) school-aged children (6-11 years old). House dust from the living room and from bedroom floors were analyzed for endotoxin and beta-glucan contents. House dust was analyzed for endotoxin content by the kinetic limulus amoebocyte lysate assay and for β-(1→3)-D-glucan by the inhibition enzyme immunoassay. The parents answered questions regarding potential determinants. We found geometric means 187.5 mg/m² for dust. According to statistical values, the endotoxin geometric mean was 13.86×103 EU/g for the control group and 6.16×103 EU/g for the asthma group. As a result, the amount of bacterial endotoxin was measured at a higher level in the homes of children without asthma. The geometric mean for beta-glucan was 46.52 µg/g and 44.39 µg/g for asthma and control groups, respectively. No associations between asthma and microbial agents were observed in Turkish children. High correlations (r > 0.75) were found between floor dust and endotoxin loads, while endotoxin and β-(1→3)-D-glucan concentrations were not correlated. The type of flooring (hard-surface or textile) was the strongest determinant for loads of floor dust and concentrations of endotoxin. Water damage and dampness at home were determinants of β-(1→3)-D-glucan concentrations. Endotoxin and β-(1→3)-D-glucan concentrations in Turkish house dust might lower than concentrations seen in other European countries.

Keywords: indoor air quality, asthma, microbial pollutants, case-control

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Authors: S. S. L. Andrade, E. A. Souza, L. C. L. Santos, C. Moraes, A. K. C. L. Lobato

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Biodiesel may be produced through transesterification reaction (or alcoholysis), that is the transformation of a long chain fatty acid in an alkyl ester. This reaction can occur in the presence of acid catalysts, alkali, or enzyme. Currently, for industrial processes, biodiesel is produced by alkaline route. The alkali most commonly used in these processes is hydroxides and methoxides of sodium and potassium. In this work, biodiesel production was conducted in two different systems. The first consisted of a batch reactor operating with a traditional washing system and the second consisted of a semi-continuous flow reactor operating with a membrane separation system. Potassium hydroxides was used as catalyst at a concentration of 1% by weight, the molar ratio oil/alcohol was 1/9 and temperature of 55 °C. Tests were performed using soybeans and palm oil and the ester conversion results were compared for both systems. It can be seen that the results for both oils are similar when using the batch reator or the semi-continuous flow reactor. The use of the semi-continuous flow reactor allows the removal of the formed products. Thus, in the case of a reversible reaction, with the removal of reaction products, the concentration of the reagents becomes higher and the equilibrium reaction is shifted towards the formation of more products. The higher conversion to ester with soybean and palm oil using the batch reactor was approximately 98%. In contrast, it was observed a conversion of 99% when using the same operating condition on a semi-continuous flow reactor.

Keywords: biodiesel, batch reactor, semi-continuous flow reactor, transesterification

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Authors: Azzam H., Abousamra N. K., Wafa A. A., Hafez M. M., El-Gilany A. H.

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Platelet activation occurs in peripheral blood of patients with rheumatic mitral stenosis (MS) and atrial fibrillation (AF) and could be related to abnormal thrombogenesis. The CD40/CD40 ligand (CD40L) which reflects platelet activation, mediate a central role in thrombotic diseases. However, the role of CD40/CD40L system in rheumatic MS with or without AF remains unclear. Expressions of CD40 on monocytes and CD40L on platelets were determined by whole blood flow cytometry and serum levels of soluble CD40L were measured by enzyme-linked immunosorbent assay in group 1 (19 patients with MS) and group 2 (20 patients with MS and AF) compared to group 3 (10 controls). Patients with groups 1 and 2 had a significant increase in expression of CD40 on monocytes (P1 and P2 = 0.000) and serum levels of sCD40L (P1 = 0.014 and P2 = 0.033, respectively), but nonsignificant increase in expression of CD40L on platelets (P1 = 0.109 and P2 = 0.060, respectively) as compared to controls. There were no significant difference in all the parameters in group 1 compared to group 2. Correlation analysis demonstrated that there was a significant direct relationship between the severity of MS and serum levels of sCD40L (r = -0.469, p = 0.043). In conclusion, rheumatic MS patients with or without AF had upregulation of the CD40/CD40L system as well as elevated sCD40L levels. The levels of sCD40L had a significantly direct relationship with the severity of MS and it was the stenotic mitral valve, not AF, that had a significant impact on platelet activation.

Keywords: CD40, CD40L, mitral stenosis, atrial fibrillation

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Authors: Masaki Ohno, Cervinia Manalo, Tetsuji Okuda, Satoshi Nakai, Wataru Nishijima

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Reverse osmosis (RO) membranes have been widely used for desalination to purify water for drinking and other purposes. Although at present most RO membranes have no resistance to chlorine, chlorine-resistant membranes are being developed. Therefore, direct chlorine treatment or chlorine washing will be an option in preventing biofouling on chlorine-resistant membranes. Furthermore, if particle accumulation control is possible by using chlorine washing, expensive pretreatment for particle removal can be removed or simplified. The objective of this study was to determine the effective hypochlorite washing condition required for controlling biofilm formation and inorganic particle accumulation on RO membrane in a continuous flow channel with RO membrane and spacer. In this study, direct chlorine washing was done by soaking fouled RO membranes in hypochlorite solution and fluorescence intensity was used to quantify biofilm on the membrane surface. After 48 h of soaking the membranes in high fouling potential waters, the fluorescence intensity decreased to 0 from 470 using the following washing conditions: 10 mg/L chlorine concentration, 2 times/d washing interval, and 30 min washing time. The chlorine concentration required to control biofilm formation decreased as the chlorine concentration (0.5–10 mg/L), the washing interval (1–4 times/d), or the washing time (1–30 min) increased. For the sample solutions used in the study, 10 mg/L chlorine concentration with 2 times/d interval, and 5 min washing time was required for biofilm control. The optimum chlorine washing conditions obtained from soaking experiments proved to be applicable also in controlling biofilm formation in continuous flow experiments. Moreover, chlorine washing employed in controlling biofilm with suspended particles resulted in lower amounts of organic (0.03 mg/cm²) and inorganic (0.14 mg/cm²) deposits on the membrane than that for sample water without chlorine washing (0.14 mg/cm² and 0.33 mg/cm², respectively). The amount of biofilm formed was 79% controlled by continuous washing with 10 mg/L of free chlorine concentration, and the inorganic accumulation amount decreased by 58% to levels similar to that of pure water with kaolin (0.17 mg/cm²) as feed water. These results confirmed the acceleration of particle accumulation due to biofilm formation, and that the inhibition of biofilm growth can almost completely reduce further particle accumulation. In addition, effective hypochlorite washing condition which can control both biofilm formation and particle accumulation could be achieved.

Keywords: reverse osmosis, washing condition optimization, hypochlorous acid, biofouling control

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Authors: T. N. M. El Sebai, A. A. Khattab, Wafaa M. Abd-El Rahim, H. Moawad

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Fungal mutant strains have produced cellulase and xylanase enzymes, and have induced high hydrolysis with enhanced of rice straw. The mutants were obtained by exposing Penicillium strain to UV-light treatments. Screening and selection after treatment with UV-light were carried out using cellulolytic and xylanolytic clear zones method to select the hypercellulolytic and hyperxylanolytic mutants. These mutants were evaluated for their cellulase and xylanase enzyme production as well as their abilities for biodegradation of rice straw. The mutant 12 UV/1 produced 306.21% and 209.91% cellulase and xylanase, respectively, as compared with the original wild type strain. This mutant showed high capacity of rice straw degradation. The effectiveness of tested mutant strain and that of wild strain was compared in relation to enhancing the composting process of rice straw and animal manures mixture. The results obtained showed that the compost product of inoculated mixture with mutant strain (12 UV/1) was the best compared to the wild strain and un-inoculated mixture. Analysis of the composted materials showed that the characteristics of the produced compost were close to those of the high quality standard compost. The results obtained in the present work suggest that the combination between rice straw and animal manure could be used for enhancing the composting process of rice straw and particularly when applied with fungal decomposer accelerating the composting process.

Keywords: rice straw, composting, UV mutants, Penicillium

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Authors: Naveen Kumar

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Microplastics, tiny plastic particles less than 5 mm in size formed by the disposal and breakdown of industrial and consumer products, have become a primary environmental concern due to their ubiquitous presence and application in the environment and their potential to cause harm to the ecosystem, wildlife and human health. In this, we study the ability of the fungus Rasamsonia emersonii IMI 393752 to degrade the rigid microplastics of Coke bottles. Microplastics were extracted from Coke bottles and incubated with Rasamsonia emersonii in Sabouraud dextrose agar media. Microplastics were pre-sterilized without altering the chemistry of microplastic. Preliminary analysis was performed by observing radial growth assessment of microplastic-containing media enriched with fungi vs. control. The assay confirmed no impedance or change in the fungi's growth pattern and rate by introducing microplastics. The degradation of the microplastics was monitored over time using microscopy and FTIR, and biodegradation/deterioration on the plastic surface was observed. Furthermore, the liquid assay was performed. HPLC and GCMS will be conducted to check the biodegradation and presence of enzyme release by fungi to counteract the presence of microplastics. These findings have important implications for managing plastic waste, as they suggest that fungi such as Rasamsonia emersonii can potentially degrade microplastics safely and effectively. However, further research to optimise the conditions for microplastic degradation by Rasamsonia emersonii and to develop strategies for scaling up the process for industrial applications will be beneficial.

Keywords: bioremediation, mycoremediation, plastic degradtion, polyethylene terephthalate

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Authors: Zahra Azizi, Ashkan Karbasi, Farzin Firouzian, Sara Soleimani Asl, Akram Ranjbar

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Background: One of the most often used herbicides in agriculture is paraquat (PQ), which is very harmful to both people and animals. Chitosan is a well-known, non-toxic polymer commonly used in preparing particles via ionotropic gelation facilitated by negatively charged agents such as sodium alginate. This study aimed to compare the effects of PQ and nanoparaquat (PQNPs) on liver function in male rats. Materials & Methods: Rats were exposed to PQ & PQNPs (4 mg/kg/day, intraperitoneally) for seven days. Then, rats were anesthetized, and serum and liver samples were collected. Later, enzymatic activities such as alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP) in serum and oxidative stress biomarkers such as lipid peroxidation (LPO), total antioxidant capacity (TAC) and total thiol groups (TTG) levels in liver tissue were measured by colorimetric methods. Also, histological changes in the liver were evaluated. Results: PQ altered the levels of ALT, AST, and ALP while inducing oxidative stress in the liver. Additionally, liver homogenates with PQ exposure had challenged LPO, TAC, and TTG levels. The severe liver damage is indicated by a significant increase in the enzyme activity of AST, ALT, and ALP in serum. According to the results of the current study, PQNPs, as compared to PQ and the control group, lowered ALT, AST, ALP, and LPO levels while increasing TAC and TTG levels. Conclusion: According to biochemical and histological investigations, PQ loaded in chitosan-alginate particles is more efficient than free PQ at reducing liver toxicity.

Keywords: paraquat, paraquat nanoparticles, liver, oxidative stress

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Authors: Rini Rahul, Manoj Kumar

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Cadmium (Cd) is a poisonous trace metal, which is responsible for excess reactive oxygen species generation (ROS) in plants, thereby adversely affecting their productivity and commercial potential. Ricinus communis (castor) is an industry-efficient non-edible bioenergy crop used for phytoremediation and re-vegetation. We have determined the total Cd content in castor genotypes and established a relationship between the Cd tolerance mechanism and physiological parameters like chlorophyll fluorescence, the total photosynthetic activity, chlorophyll and carotenoid content as well as ROS generation and malondialdehyde content. This study is an effort to comprehend the interrelation between Cd toxicity (control, 250 µM and 500 µM), proline, various ROS scavenging enzymes (anti-oxidative in nature), nicotianamine synthase (NAS) and Natural resistance-associated macrophage protein (NRAMP) gene. The antioxidant enzyme activity increased for WM hence conferring Cd toxicity in this genotype. RcNRAMP genes showed differential expression in GCH2 and WM genotypes; this can also be one of the reasons for Cd toxicity and sensitivity in WM and GCH2, respectively. The cause of pronounced Cd tolerance in WM leaves can be because of enhanced expression of RcNAS1, RcNAS2 and RcNAS3 genes. Our results demonstrate that there is an interrelation between Cd toxicity (control, 250 µM and 500 µM), proline, various ROS scavenging enzymes (anti-oxidative in nature), NAS and NRAMP gene.

Keywords: ricinus communis, cadmium, reactive oxygen species, nicotianamine synthase, NRAMP, malondialdehyde

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Authors: Evren C. Eroglu, Ridvan Arslan

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Over the course of herbal production, various pretreatments are performed and some of which have serious effect on the bioactive properties. Especially in the production of herbal tea from fresh herbals, it is considered that elapsed time from blending to last product may affect the bioactive properties and antioxidant contents. Herbal infusion is basically prepared by mixing herbs with hot water for 10-20 min. During the brewing of these herbs, it is supposed to be significant decrease in the antioxidant and phenolics content. The first aim of this study was to evaluate the changes of vitamin C (VitC), total phenolic content (TPC) and antioxidant contents (AO) of two brassica varieties (brussel sprouts and white head cabbage) with different holding time after blending. Second aim of this study was to understand the effect of herbal infusion on VitC, TPC and AO contents. In this study, fresh samples were subjected to 0-30 min holding time after blending. Then, samples was immediately taken to -80 °C and freeze drying process was performed. Herbal infusion was performed for 20 minutes. According to results, VitC contents in brussel sprouts was not changed significantly (p=0.12). However, there was a significant decreasing of VitC content in cabbage sample (p=0.034). 20 min of brewing caused a significant decrement in VitC of brussel sprouts by approximately 76% (1071 ppm dw), while decline in cabbage VitC content was 87% (531 ppm dw). AO and TPC values of unprocessed cabbage control sample (13791.87 ppm FeSO4·7H2O eq. dw and 5301.85 ppm gallic acid eq. dw) were higher than brussel sprouts control samples (11571.75 ppm FeSO4·7H2O dw and 5202.76 ppm, respectively). The change in AO and TPC of both brussel sprouts and cabbage samples were not statistically significant at the end of 30 minutes holding time (p=0.24 and p=0.38). After 20 minutes of brewing, AO content in brussel sprouts significantly decreased by 44% (p ˂0.05). Although, the decreasing of AO in white head cabbage was statistically important (p=0.034), decreasing was just 8%. TPC values were found to decrease by 54% in cabbage, while it was 35% in brussel sprouts after herbal infusion. It was observed that 30 min holding time had no statistically important effect on TPC values of both cabbage and brussel sprouts. As a conclusion, herbal infusion has more or less effect on VitC, TPC and AO contents of samples. Therefore, it is important to decrease brewing time. Another result was that there were no significant differences in TPC and AO content of both samples when holding samples 30 min outside after blending. However, this process had significant effect on VitC content of white head cabbage.

Keywords: Antioxidant content, brussel sprouts, herbal infusion, total phenolic content, white head cabbage, vitamin c

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Authors: Rizky Kusuma Cahyani

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Hepatitis B is a liver inflammatory disease caused by hepatitis B virus (HBV). This infection can be prevented by vaccination which contains HBV surface protein (sHBsAg). However, vaccine supply is limited. Several attempts have been conducted to produce local sHBsAg. However, the purity degree and protein yield are still inadequate. Therefore optimization of HBsAg purification steps is required to obtain high yield with better purification fold. In this study, optimization of purification was done in 2 steps, precipitation using variation of NaCl concentration (0,3 M; 0,5 M; 0,7 M) and PEG (3%, 5%, 7%); ion exchange chromatography (IEC) using NaCl 300-500 mM elution buffer concentration.To determine HBsAg protein, bicinchoninic acid assay (BCA) and enzyme-linked immunosorbent assay (ELISA) was used in this study. Visualization of HBsAg protein was done by SDS-PAGE analysis. Based on quantitative analysis, optimal condition at precipitation step was given 0,3 M NaCl and PEG 3%, while in ion exchange chromatography step, the optimum condition when protein eluted with NaCl 500 mM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicates that the presence of protein HBsAg with a molecular weight of 25 kDa (monomer) and 50 kDa (dimer). The optimum condition for purification of sHBsAg produced in Pichia pastoris gave a yield of 47% and purification fold 17x so that it would increase the production of hepatitis B vaccine to be more optimal.

Keywords: hepatitis B virus, HBsAg, hepatitis B surface antigen, Pichia pastoris, purification

Procedia PDF Downloads 135

Authors: Jianfang Wang, Xianzhe Chen, Zhuoliang Liu, Cheng-An Tao, Yujiao Li

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Organophosphorus (OPs) pesticide used as insecticides are widely used in agricultural pest control, household and storage deworming. The detection of pesticides needs more simple and efficient methods. One of the best ways is to make electrochemical biosensors. In this paper, an electrochemical enzyme biosensor based on acetylcholine esterase (AChE) was constructed, and its sensing properties and sensing mechanisms were studied. Reduced graphene oxide-polydopamine complexes (RGO-PDA), gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs) were prepared firstly and composited with AChE and chitosan (CS), then fixed on the glassy carbon electrode (GCE) surface to construct the biosensor GCE/RGO-PDA-AuNPs-AgNPs-AChE-CS by one-pot method. The results show that graphene oxide (GO) can be reduced by dopamine (DA) and dispersed well in RGO-PDA complexes. And the composites have a synergistic catalysis effect and can improve the surface resistance of GCE. The biosensor selectively can detect acetylcholine (ACh) and OPs pesticide with good linear range and high sensitivity. The performance of the biosensor is affected by the ratio and adding ways of AChE and the adding of AuNPs and AChE. And the biosensor can achieve a detection limit of 2.4 ng/L for methyl parathion and a wide linear detection range of 0.02 ng/L ~ 80 ng/L, and has excellent stability, good anti-interference ability, and excellent preservation performance, indicating that the sensor has practical value.

Keywords: acetylcholine esterase, electrochemical biosensor, nanoparticles, organophosphates, reduced graphene oxide

Procedia PDF Downloads 95