Search results for: heart-type fatty acid binding protein

Authors: A. Bara, D. Barkat

Abstract:

Liquid-liquid extraction is one of the most useful techniques for selective removal and recovery of metal ions from aqueous solutions, applied in purification processes in numerous chemical and metallurgical industries. In this work, The liquid-liquid extraction of cobalt (II) and copper (II) from aqueous solution by capric acid (HL) in chloroform at 25°C has been studied. Our interest in this paper is to study the effect of concentration of capric acid on the extraction of Co(II) and Cu(II) to see the complexes could be formed in the organic phase using various concentration of capric acid. The extraction of cobalt (II) and copper (II) is extracted as the complex CoL2 (HL )2, CuL2 (HL)2.

Keywords: capric acid, Cobalt(II), copper(II), liquid-liquid extraction

Procedia PDF Downloads 421

Authors: Ashrafosadat Mousavi Laer, Elaheh Moravej

Abstract:

The art of binding was simple and elementary at the beginning of Islam. This art thrived gradually and continued its development as an independent art. Identification of the binding techniques and used materials in covers and investigation of the arrays give us indexes for the better identification of different doctrines and methods of that time. The catalogers of the manuscripts usually pay attention to four items: gender, color, art elegances, injury, and exquisiteness of the cover. The criterion for classification of the covers is their art nature and gender. 15th century AD (9th century AH) was the period of the binding art development in which the most beautiful covers were produced by the so-called method of ‘burning’. At 16th century AD (10th century AH), in Safavid era, art changed completely and a fundamental evolution occurred in the technique and method of binding. The greatest change in this art was the extensive use of stamp that was made mostly of steel and copper. Theses stamps were presses against leather. These covers were called ‘beat’. In this paper, writing and bookbinding of about 32 Shahnamehs of Safavid era available in the Iranian libraries and museums are studied. An analytical-statistical study shows that four methods have been used including beat, burning, mosaic, and oily. 69 percent of the covers of these copies are cardboards with a leathery coating (goatskin) and have been produced by burning and beat methods. Its reasons are that these two methods have been common methods in Safavid era and performing them was only feasible on leather and the most desirable and commonly used leather of that time was goatskin which was the best option for cover legend durability and preserving the book and it was more durable because it had been made of goat skin. In addition, it had prepared a suitable opportunity for the binding artist’s creativity and innovation.

Keywords: Shahnameh, Safavid era, bookbinding, beat cover, burning cover

Procedia PDF Downloads 223

Authors: Azhar Jabareen, Mahmoud Huleihel

Abstract:

BRCA-1 is a multifunctional tumor suppressor, whose expression is activated by the estrogen (E2)-liganded ERα receptor. The activated ERα is a transcriptional factor which activates various genes either by direct binding to the DNA at E2-responsive elements (EREs) and indirectly associated with a range of alternative non-ERE elements. Interference with BRCA-1 expression and/or functions leads to high risk of breast or/and ovarian cancer. Our lab investigated the involvement of Human T-cell leukemia Virus Type 1 (HTLV-1) in breast cancer, since HTLV-1 Tax was found to strongly inhibit BRCA-1 expression. In addition, long exposure of 12-O-tetradecanoylphorbol-13-acetate (TPA), which is one of the stress-inducing agents activated the HTLV-1 promoter. So here the involvement of TPA in breast cancer had been examined by testing the effect of TPA on BRCA-1 and ERE expression. The results showed that TPA activated both BRCA-1 and ERE expression. In the 12 hours TPA activated the tow promoters more than others time, and after 24 hours the level of the tow promoters was decreased. Tax inhibited BRCA-1 expression but did not succeed to inhibit the effect of TPA. Then the activation of the two promoters was not through ERα pathway because TPA had no effect on ERα binding to the two promoters of the BRCA-1 and ERE. Also, the activation was not via nuclear factor kappa B (NF-κB) pathway because when the inhibitory of NF-κB had been added to the TPA, it still activated the tow promoters. However, it seems that 53BP1 may be involved in TPA activation of these promoters because ectopic high expression of 53BP1 significantly reduced the TPA activity. In addition, in the presence of Bisindolylmaleimide-I (BI)- the inhibitor of Protein Kinase C (PKC)- there was no activation for the two promoters, so the PKC is agonized BRCA-1 and ERE activation.

Keywords: BRCA-1, ERE, HTLV-1, TPA

Procedia PDF Downloads 230

Authors: Neslihan Taskale Karatug, Mustafa Akcelik

Abstract:

Some literature data show that misL protein play a role on host immune response formed against Salmonella Typhimurium. The aim of the present study is to learn the role of the protein in S. Typhimurium pathogenicity. To describe certain functions of the protein, primarily recombinant misL protein was produced and purified. PCR was performed using a primer set targeted to passenger domain of the misL gene on S. Typhimurium LT2 genome. Amplicon and pet28a vector were enzymatically cleaved with EcoRI and NheI. The digested DNA materials were purified with High Pure PCR Product Purification Kit. The ligation reaction was achieved with the pure products. After preparation of competent Escherichia coli Dh5α, ligation mix was transformed into the cell by electroporation. To confirm the existence of insert gene, recombinant plasmid DNA of Dh5α was isolated with high pure plasmid DNA kit. Proved the correctness of recombinant plasmid was electroporated to BL21. The cell was induced by IPTG. After induction, the presence of recombinant protein was checked by SDS-PAGE. The recombinant misL protein was purified using HisPur Ni-NTA spin colon. The pure protein was shown by SDS-PAGE and western blot immünoassay. The concentration of the protein was measured BCA Protein Assay kit. In the wake of ligation with digested products (2 kb misL and 5.4 kb pet28a) visualised on gel size of the band was about 7.4 kb and was named as pNT01. The pNT01 recombinant plasmid was transformed into Dh5α and colonies were chosen in selective medium. Plasmid DNA isolation from them was carried out. PCR was achieved on the pNT01 to check misL and 2 kb band was observed on the agarose gel. After electroporation of the plasmid and induction of the cell, 68 kDa misL protein was seen. Subsequent to the purification of the protein, only a band was observed on SDS-PAGE. Association of the pure protein with anti-his antibody was verified by the western blot assay. The concentration of the pure misL protein was determined as 345 μg/mL. Production of polyclonal antibody will be achieved by using the obtained pure recombinant misL protein as next step. The role of the protein will come out on the immune system together some assays.

Keywords: cloning, Escherichia coli, recombinant protein purification, Salmonella Typhimurium

Procedia PDF Downloads 371

Authors: Mina Kargozari, Isabel Revilla Martin, Ángel A. Carbonell-Barrachina

Abstract:

Different formulations of Turkish fermented sausages (sucuks) prepared with camel meat-hump (CH), camel meat-beef fat (CB), beef-hump (BH) and beef-beef fat (BB), were characterized. The sausages were analytically compared to determine differences in proximate composition and total cholesterol content (TCC), quality parameters such as fatty acids profile and fat quality characteristics, and lipid oxidation parameters including peroxide value, thiobarbituric acid-reactive substances (TBARS) and resulted carbonyl compounds. The PUFAs/SFAs ratio was higher in CB and BB samples than CH and BH (p<0.05). The higher calculated atherogenic and thrombogenic indexes (AI and TI) were obtained from the samples made with hump (p< 0.05) as a result of high amounts of their SFAs. The CH sausages contained high amount of total fat (p<0.05) among all samples. The CB sucuks exhibited the highest protein content and the lowest TCC and rancidity at the end of ripening (p<0.05). The TBARS results showed that beef fat samples were more susceptible to lipid oxidation. Moreover, no significant difference (p<0.05) was observed for the values of short aldehydes among the sucuk samples excepting nonanal. This study demonstrated that supplementing camel meat for the production of dry-fermented sausage resulted in high quality products with good functional and nutritional characteristics.

Keywords: fermented sausages, quality properties, SPME, total cholesterol content

Procedia PDF Downloads 303

Authors: Vita Sterna, Sanita Zute, Inga Jansone, Linda Brunava, Inara Kantane

Abstract:

Grains, including oats (Avena sativa L.), have been recognized functional foods, because provide beneficial effect on the health of the consumer and decrease the risk of various diseases.Oats are good source of soluble fibre, essential amino acids, unsaturated fatty acids, vitamins and minerals. Oat breeders have developed oat varieties and improved yielding ability potential of oat varieties. Therefore, the aim of investigation was to analyze the composition of perspective oat varieties and breeding lines grains grown in different conditions and evaluate functional properties. In the studied samples content of protein, starch, β - glucans, total dietetic fibre, composition of amino acids and vitamin E were determined. The results of analysis showed that protein content depending of varieties ranged 9.70 –17.30% total dietary fibre 13.66-30.17 g100g-1, content of β-glucans 2.7-3.5 g100g-1, amount of vitamin E (α-tocopherol) determined from 4 to 9.9 mg kg-1. The sum of essential amino acids in oat grain samples were determined from 31.63 to 54.90 gkg-1. Concluded that amino acids composition of husked and naked oats grown in organic or conventional conditions is close to optimal.

Keywords: dietetic fibre, amino acids, scores, nutrition value

Procedia PDF Downloads 480

Authors: Nurul Aqilah Mohd Zaini, Afroditi Chatzifragkou, Dimitris Charalampopoulos

Abstract:

D-Lactic acid production is gaining increasing attention due to the thermostable properties of its polymer, Polylactic Acid (PLA). In this study, D-lactic acid was produced in microbial cultures using Lactobacillus coryniformis subsp. torquens as D-lactic acid producer and hydrolysates of Dried Distillers Grains with Solubles (DDGS) as fermentation substrate. Prior to fermentation, DDGS was first alkaline pretreated with 5% (w/v) NaOH, for 15 minutes (121oC/ ~16 psi). This led to the generation of DDGS solid residues, rich in carbohydrates and especially cellulose (~52%). The carbohydrate-rich solids were then subjected to enzymatic hydrolysis with Accellerase® 1500. For Separate Hydrolysis and Fermentation (SHF), enzymatic hydrolysis was carried out at 50oC for 24 hours, followed by fermentation of D-lactic acid at 37oC in controlled pH 6. The obtained hydrolysate contained 24 g/l glucose, 5.4 g/l xylose and 0.6 g/l arabinose. In the case of Simultaneous Saccharification and Fermentation (SSF), hydrolysis and fermentation were conducted in a single step process at 37oC in pH 5. The enzymatic hydrolysis of DGGS pretreated solids took place mostly during lag phase of L. coryniformis fermentation, with only a small amount of glucose consumed during the first 6 h. When exponential phase was started, glucose generation reduced as the microorganism started to consume glucose for D-lactic acid production. Higher concentrations of D-lactic acid were produced when SSF approach was applied, with 28 g/l D-lactic acid after 24 h of fermentation (84.5% yield). In contrast, 21.2 g/l D-lactic acid were produced when SHF was used. The optical pu rity of D-lactic acid produced from both experiments was 99.9%. Besides, approximately 2 g/l acetic acid was also generated due to lactic acid degradation after glucose depletion in SHF. SSF was proved an efficient towards DDGS ulilisation and D-lactic acid production, by reducing the overall processing time, yielding sufficient D-lactic acid concentrations without the generation of fermentation by-products.

Keywords: DDGS, alkaline pretreatment, SSF, D-lactic acid

Procedia PDF Downloads 318

Authors: Taliman Nisar Ahmad, Hirofume Saneoka

Abstract:

A pot experiment was conducted to investigate the effect of phosphorus (P) and potassium (K) nutrition on grain yield, phytic acid and grain quality of high-phytate (Akimaro) and low-phytate line. Phosphorus and potassium were applied as; P₁ (20 kg ha⁻¹) and P₂ (100 kg ha⁻¹), same as K₁ (20 kg ha⁻¹) and K₂ (100 kg ha⁻¹), respectively. Low-phytate soybean had the highest grain yield, and 75% increase was observed compared to the high-phytate under same treatments. Highly significant differences of seed phytate P were observed in both cultivars, and the phytate P in high-phytate was found 39% higher than low-phytate, whereas no significant differences observed in response to P and K treatment. Percentage of phytate P from total P in seeds was 28 to 35% in low-phytate and 72 to 81% in high-phytate in different treatments. The lipid content in low-phytate was found lowered compared to that of high-phytate. Crude protein in grains was also found significantly higher in PK combined. No significant difference was observed in seed calcium (Ca), magnesium (Mg), and Zinc (Zn) in different treatments, but high-phytate showed 87% increase in seed Ca and 76% of Mg compared to low-phytate; however, low-phytate showed 82% increase in Zn content over high-phytate. The result illustrates that low-phytate soybean achieved higher grain yield and grain Pi in response to increased P and K nutrition. To achieve higher yield and quality seeds from the low-phytate soybean, it is recommended that proper phosphorus and potassium nutrition to be applied suggested in this study.

Keywords: phytic acid, low-phytate soybean, high-phytate soybean, P and K nutrition, protein content, soybean

Procedia PDF Downloads 117

Authors: Debashree Ghosh, Preethi Sridhar, Shloka Atul Dhavle

Abstract:

The judicious and economic consumption of energy for sustainable growth and development is nowadays a thing of primary importance; Phase Change Materials (PCM) provide an ingenious option of storing energy in the form of Latent Heat. Energy storing mechanism incorporating phase change material increases the efficiency of the process by minimizing the difference between supply and demand; PCM heat exchangers are used to storing the heat or non-convectional energy within the PCM as the heat of fusion. The experimental study evaluates the effect of thermo-physical properties, variation in inlet temperature, and flow rate on charging period of a coiled heat exchanger. Secondly, a numerical study is performed on a PCM double pipe heat exchanger packed with two different PCMs, namely, RT50 and Fatty Acid, in the annular region. In this work, the simulation of charging of paraffin wax (RT50) using water as high-temperature fluid (HTF) is performed. Commercial software Ansys-Fluent 15 is used for simulation, and hence charging of PCM is studied. In the Enthalpy-porosity model, a single momentum equation is applicable to describe the motion of both solid and liquid phases. The details of the progress of phase change with time are presented through the contours of melt-fraction, temperature. The velocity contour is shown to describe the motion of the liquid phase. The experimental study revealed that paraffin wax melts with almost the same temperature variation at the two Intermediate positions. Fatty acid, on the other hand, melts faster owing to greater thermal conductivity and low melting temperature. It was also observed that an increase in flow rate leads to a reduction in the charging period. The numerical study also supports some of the observations found in the experimental study like the significant dependence of driving force on the process of melting. The numerical study also clarifies the melting pattern of the PCM, which cannot be observed in the experimental study.

Keywords: latent heat storage, charging period, discharging period, coiled heat exchanger

Procedia PDF Downloads 98

Authors: Feng Ju Chuang, Yu Wen Wang, Tai Jung Hsieh, Shyh Ming Kuo

Abstract:

Polylactic acid is a synthetic polymer with good biocompatibility and degradability, is widely used in clinical applications. In this study, we utilized Polylactic acid particles as stimulants for macrophages and the collagen synthesis of co-cultured fibroblasts was evaluated. The results indicated that Polylactic acid particles were nontoxic to cells from 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. No obvious inflammation effect was observed (under the PLLA concentration of 1 mg/mL) after 24-h co-culture of Raw264.7 and NIH3T3 cells (from TNF-α assay). The addition of PLLA particles to the Raw264.7 and NIH3T3 co-cultures increased the synthesis of collagen, the highest collagen synthesis from the fibroblast was the 0.2 mg/mL (approximately 60% increased as compared with without addition Polylactic acid particles). Moreover, a co-axial atomization delivery device was used to percutaneously introduce Polylactic acid particles into the dermis layer and stimulating macrophages to secrete growth factors promoting fibroblasts to produce collagen. The preliminary results demonstrated the synthesis of collagen was increased mildly after the introduction of Polylactic acid particles for 28-d post implantation. The Polylactic acid particles could be successfully introduced into the dermis layer from H&E staining examination, however, the optimum concentration of Polylactic acid particles and the time-period for collagen synthesis still need to be evaluated.

Keywords: collagen synthesis, macrophage, NIH3T3 cells, polylactic acid particles

Procedia PDF Downloads 88

Authors: Abdul Soofi, Katherine Wolf, Egon Ranghini, Gregory Dressler

Abstract:

Obesity and its associated complications, such as insulin resistance and non-alcoholic fatty liver disease, are reaching epidemic proportions. In mice, the TGF-β superfamily is implicated in the regulation of white and brown adipose tissues differentiation. The Kielin/Chordin-like Protein (KCP) is a secreted regulator of the TGF-β superfamily pathways that can inhibit both TGF-β and Activin signals while enhancing the Bone Morphogenetic protein (BMP) signaling. However, the effects of KCP on metabolism and obesity have not been studied in animal models. Thus, we examined the effects of KCP loss or gain of function in mice that were maintained on either a regular or a high fat diet. Loss of KCP sensitized mice to obesity and associated complications such as hepatic steatosis and glucose intolerance. In contrast, transgenic mice that expressed KCP in the kidney, liver and adipose tissues were resistant to developing high fat diet induced obesity and had significantly reduced white adipose tissue. KCP over-expression was able to shift the pattern of Smad signaling in vivo, to increase the levels of P-Smad1 and decrease P-Smad3, resulting in resistance to high fat diet induced hepatic steatosis and glucose intolerance. In aging mice, loss of KCP promoted liver pathology even when mice were fed a normal diet. The data demonstrate that shifting the TGF-β superfamily signaling with a secreted inhibitor or enhancer can alter the physiology of adipose tissue to reduce obesity and can inhibit the initiation and progression of hepatic steatosis to significantly reduce the effects of high fat diet induced metabolic disease.

Keywords: adipose tissue, KCP, obesity, TGF-β, BMP, hepatic steatosis, metabolic syndrome

Procedia PDF Downloads 338

Authors: Reza Samsami

Abstract:

The aim of the study was to degradation of polycyclic aromatic hydrocarbons (PAHs) by proxy-acid method. The amounts of PAHs were determined in a silty-clay soil sample of an aged oil refinery field in Abadan, Iran. Proxy-acid treatment method was investigated. The results have shown that the proxy-acid system is an effective method for degradation of PAHs. The results also demonstrated that the number of fused aromatic rings have not significant effects on PAH removal by proxy-acid method. The results also demonstrated that the number of fused aromatic rings have not significant effects on PAH removal by proxy-acid method.

Keywords: proxy-acid treatment, silty-clay soil, PAHs, degradation

Procedia PDF Downloads 249

Authors: I. Sarenkova, I. Ciprovica, I. Cinkmanis

Abstract:

Lactobionic acid is a disaccharide formed from gluconic acid and galactose, and produced by oxidation of lactose. Productivity of lactobionic acid by microbial synthesis can be affected by various factors, and one of them is a presence of potassium, magnesium and manganese ions. In order to extend lactobionic acid production efficiency, it is necessary to increase the yield of lactobionic acid by optimising the fermentation conditions and available substrates for Pseudomonas taetrolens growth. The object of the research was to determinate the application of K₂HPO₄, MnSO₄, MgSO₄ × 7H₂O salts in different concentration for effective lactose oxidation to lactobionic acid by Pseudomonas taetrolens. Pseudomonas taetrolens NCIB 9396 (NCTC, England) and Pseudomonas taetrolens DSM 21104 (DSMZ, Germany) were used for the study. The acid whey was used as the study object. The content of lactose in whey samples was determined using MilcoScan^TM Mars (Foss, Denmark) and high performance liquid chromatography (Shimadzu LC 20 Prominence, Japan). The content of lactobionic acid in whey samples was determined using the high performance liquid chromatography. The impact of studied salts differs, Mn²⁺ and Mg²⁺ ions enhanced fermentation instead of K⁺ ions. Results approved that Mn²⁺ and Mg²⁺ ions are necessary for Pseudomonas taetrolens growth. The study results will help to improve the effectiveness of lactobionic acid production with Pseudomonas taetrolens NCIB 9396 and DSM 21104.

Keywords: lactobionic acid, lactose oxidation, Pseudomonas taetrolens, whey

Procedia PDF Downloads 138

Authors: Vagolu S. Krishna, Shan Zheng, Estharla M. Rekha, Luke W. Guddat, Dharmarajan Sriram

Abstract:

Tuberculosis (TB) remains a major threat to human health. This due to the fact that current drug treatments are less than optimal as well as the rising occurrence of multi drug-resistant and extensively drug-resistant strains of the etiological agent, Mycobacterium tuberculosis (Mt). Given the wide-spread significance of this disease, we have undertaken a design and evaluation program to discover new anti-TB drug leads. Here, our attention is focused on ketol-acid reductoisomerase (KARI), the second enzyme in the branched-chain amino acid biosynthesis pathway. Importantly, this enzyme is present in bacteria but not in humans, making it an attractive proposition for drug discovery. In the present work, we used high-throughput virtual screening to identify seventeen potential inhibitors of KARI using the Birla Institute of Technology and Science in-house database. Compounds were selected based on high docking scores, which were assigned as the result of favourable interactions between the compound and the active site of KARI. The Ki values for two leads, compounds 14 and 16 are 3.71 and 3.06 µM, respectively for Mt KARI. To assess the mode of binding, 100 ns molecular dynamics simulations for these two compounds in association with Mt KARI were performed and showed that the complex was stable with an average RMSD of less than 2.5 Å for all atoms. Compound 16 showed an MIC of 2.06 ± 0.91 µM and a 1.9 fold logarithmic reduction in the growth of Mt in an infected macrophage model. The two compounds exhibited low toxicity against murine macrophage RAW 264.7 cell lines. Thus, both compounds are promising candidates for development as an anti-TB drug leads.

Keywords: ketol-acid reductoisomerase, macrophage, molecular docking and dynamics, tuberculosis

Procedia PDF Downloads 107

Authors: Hadriana Bansi, Elizabeth Wina, Toto Toharmat

Abstract:

Acacia villosa is thornless shrub legume which contents high crude protein. However, the utilization of A. villosa as ruminant feed is limited by its secondary compounds. The aim of this article is to find out the maximum of substitution A. villosa in sheep ration. The nutritional evaluation consisted of in vitro two stages, in vivo, and in vitro gas production trials. The secondary compounds of A. villosa also were analyzed. Evaluating digestibility of increasing level of substitution A. villosa replacing Pennisetum purpureum was using in vitro two stages. The substitution of 30% A. villosa was compared to 100% P. purpureum by in vitro gas production technique and in vivo digestibility. The results of two stages in vitro showed that total phenol, condensed tannin, and non-protein amino acid (NPAA) were high. Substitution 15% A. villosa reached the highest digestibility for both dry matter (DM) and crude protein (CP) which were 67% and 86% respectively, but it was shown that DM and CP digestibility of substitution 30% of A. villosa was still high which were 61.82% and 75-67% respectively. The pattern of gas production showed that first 8 hours total gas production substitution of 30% A. villosa was higher than 100% P. purpureum and declined after 10 hours incubation. In vivo trials showed that substitution of 30% A. villosa significantly increased CP intake, CP digestibility, and nitrogen retention. It can be concluded that substitution A. villosa until 30% still gave the good impact even though it has high secondary compounds.

Keywords: Acacia villosa, digestibility, gas production, secondary compounds

Procedia PDF Downloads 142

Authors: Ashima Sharma

Abstract:

Human Serum Albumin (HSA)- one of the most demanded therapeutic proteins with immense biotechnological applications- is a large multidomain protein containing 17 disulfide bonds. The current source of HSA is human blood plasma which is a limited and unsafe source. Thus, there exists an indispensable need to promote non-animal derived recombinant HSA (rHSA) production. Escherichia coli is one of the most convenient hosts which had contributed to the production of more than 30% of the FDA approved recombinant pharmaceuticals. It grows rapidly and reaches high cell density using inexpensive and simple substrates. E. coli derived recombinant products have more economic potential as fermentation processes are cheaper compared to the other expression hosts. The major bottleneck in exploiting E. coli as a host for a disulfide-rich multidomain protein is the formation of aggregates of overexpressed protein. The majority of the expressed HSA forms inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA from inclusion bodies is not preferred because it is difficult to obtain a large multidomain disulfide bond rich protein like rHSA in its functional native form. Purification is tedious, time-consuming, laborious and expensive. Because of such limitations, the E. coli host system was neglected for rHSA production for the past few decades despite its numerous advantages. In the present work, we have exploited the capabilities of E. coli as a host for the enhanced functional production of rHSA (~60% of the total expressed rHSA in the soluble fraction). Parameters like intracellular environment, temperature, induction type, duration of induction, cell lysis conditions etc. which play an important role in enhancing the level of production of the desired protein in its native form in vivo have been optimized. We have studied the effect of assistance of different types of exogenously employed chaperone systems on the functional expression of rHSA in the E. coli host system. Different aspects of cell growth parameters during the production of rHSA in presence and absence of molecular chaperones in E. coli have also been studied. Upon overcoming the difficulties to produce functional rHSA in E. coli, it has been possible to produce significant levels of functional protein through engineering the biological system of protein folding in the cell, the E. coli-derived rHSA has been purified to homogeneity. Its detailed physicochemical characterization has been performed by monitoring its conformational properties, secondary and tertiary structure elements, surface properties, ligand binding properties, stability issues etc. These parameters of the recombinant protein have been compared with the naturally occurring protein from the human source. The outcome of the comparison reveals that the recombinant protein resembles exactly the same as the natural one. Hence, we propose that the E. coli-derived rHSA is an ideal biosimilar for human blood plasma-derived serum albumin. Therefore, in the present study, we have introduced and promoted the E. coli- derived rHSA as an alternative to the preparation from a human source, pHSA.

Keywords: recombinant human serum albumin, Escherichia coli, biosimilar, chaperone assisted protein folding

Procedia PDF Downloads 195

Authors: Aliakbar Sayadi, Thomas R. Neitzert, G. Charles Clifton, Min Cheol Han

Abstract:

The present study aims to assess the performance of vermiculite concrete containing poly-lactic acid beads as an eco-friendly aggregate. Vermiculite aggregate was replaced by poly-lactic acid in percentages of 0%, 20%, 40%, 60% and 80%. Mechanical and thermal properties of concrete were investigated. Test results indicated that the inclusion of poly-lactic acid decreased the PH value of concrete and all the poly-lactic acid particles were dissolved due to the formation of sodium lactide and lactide oligomers when subjected to the high alkaline environment of concrete. In addition, an increase in thermal conductivity value of concrete was observed as the ratio of poly-lactic acid increased. Moreover, a set of equations was proposed to estimate the water-cement ratio, cement content and water absorption ratio of concrete.

Keywords: poly-lactic acid (PLA), vermiculite concrete, eco-friendly, mechanical properties

Procedia PDF Downloads 384

Authors: Dominika Pihikova, Stefan Belicky, Tomas Bertok, Roman Sokol, Petra Kubanikova, Jan Tkac

Abstract:

Since aberrant glycosylation is frequently accompanied by both physiological and pathological processes in a human body (cancer, AIDS, inflammatory diseases, etc.), the analysis of tumor-associated glycan patterns have a great potential for the development of novel diagnostic approaches. Moreover, altered glycoforms may assist as a suitable tool for the specificity and sensitivity enhancement in early-stage prostate cancer diagnosis. In this paper we discuss the construction and optimization of ultrasensitive sandwich biosensor platform employing lectin as glycan-binding protein. We focus on the immunoassay development, reduction of non-specific interactions and final glycoprofiling of human serum samples including both prostate cancer (PCa) patients and healthy controls. The fabricated biosensor was measured by label-free electrochemical impedance spectroscopy (EIS) with further lectin microarray verification. Furthermore, we analyzed different biosensor interfaces with atomic force microscopy (AFM) in nanomechanical mapping mode showing a significant differences in the altitude. These preliminary results revealing an elevated content of α-2,3 linked sialic acid in PCa patients comparing with healthy controls. All these experiments are important step towards development of point-of-care devices and discovery of novel glyco-biomarkers applicable in cancer diagnosis.

Keywords: biosensor, glycan, lectin, prostate cancer

Procedia PDF Downloads 354

Authors: Akimitsu Kugimiya, Kouhei Iwato, Toru Saito, Jiro Kohda, Yasuhisa Nakano, Yu Takano

Abstract:

The measurement of amino acid content is reported to be useful for the diagnosis of several types of diseases, including lung cancer, gastric cancer, colorectal cancer, breast cancer, prostate cancer, and diabetes. The conventional detection methods for amino acid are high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS), but they have several drawbacks as the equipment is cumbersome and the techniques are costly in terms of time and costs. In contrast, biosensors and biosensing methods provide more rapid and facile detection strategies that use simple equipment. The authors have reported a novel approach for the detection of each amino acid that involved the use of aminoacyl-tRNA synthetase (aaRS) as a molecular recognition element because aaRS is expected to a selective binding ability for corresponding amino acid. The consecutive enzymatic reactions used in this study are as follows: aaRS binds to its cognate amino acid and releases inorganic pyrophosphate. Hydrogen peroxide (H₂O₂) was produced by the enzyme reactions of inorganic pyrophosphatase and pyruvate oxidase. The Trinder’s reagent was added into the reaction mixture, and the absorbance change at 556 nm was measured using a microplate reader. In this study, an amino acid-sensing method using histidyl-tRNA synthetase (HisRS; histidine-specific aaRS) as molecular recognition element in combination with the Trinder’s reagent spectrophotometric method was developed. The quantitative performance and selectivity of the method were evaluated, and the optimal enzyme reaction and detection conditions were determined. The authors developed a simple and rapid method for detecting histidine with a combination of enzymatic reaction and spectrophotometric detection. In this study, HisRS was used to detect histidine, and the reaction and detection conditions were optimized for quantitation of these amino acids in the ranges of 1–100 µM histidine. The detection limits are sufficient to analyze these amino acids in biological fluids. This work was partly supported by Hiroshima City University Grant for Special Academic Research (General Studies).

Keywords: amino acid, aminoacyl-tRNA synthetase, biosensing, enzyme reaction

Procedia PDF Downloads 267

Authors: Joan O. Ogundele, Aladesanmi A. Oshodi, Adekunle I. Amoo

Abstract:

Abstract In-vitro multi-enzyme protein digestibility (IVMPD) and some anti-nutritional factors (ANF) of five melon (egusi) seed flours (MSF) and their protein isolates (PI) were carried out. Their PI have potentials comparable to that of soya beans. It is important to know the IVMPD and ANF of these protein sources as to ensure their safety when adapted for use as alternate protein sources to substitute for cow milk, which is relatively expensive in Nigeria. Standard methods were used to produce PI of Citrullus colocynthis, Citrullus vulgaris, African Wine Kettle gourd (Lageneria siceraria I), Basket Ball gourd (Lagenaria siceraria II) and Bushel Giant Gourd (Lageneria siceraria III) seeds and to determine the ANF and IVMPD of the MSF and PI unheated and at 37oC. Multi-enzymes used were trypsin, chymotrypsin and peptidase. IVMPD of MSF ranged from (70.67±0.70) % (C. vulgaris) to (72.07± 1.79) % (L.siceraria I) while for their PI ranged from 74.33% (C.vulgaris) to 77.55% (L.siceraria III). IVMPD of the PI were higher than those of MSF. Heating increased IVMPD of MSF with average value of 79.40% and those of PI with average of 84.14%. ANF average in MSF are tannin (0.11mg/g), phytate (0.23%). Differences in IVMPD of MSF and their PI at different temperatures may arise from processing conditions that alter the release of amino acids from proteins by enzymatic processes. ANF in MSF were relatively low, but were found to be lower in the PI, therefor making the PI safer for human consumption as an alternate source of protein.

Keywords: Anti-nutrients, Enzymatic protein digestibility, Melon (egusi)., Protein Isolates.

Procedia PDF Downloads 94

Authors: Illyas Md Isa, Maryam Musfirah Che Sobry, Mohamad Syahrizal Ahmad, Nurashikin Abd Azis

Abstract:

A single-walled carbon nanotube (SWCNT) that has been modified with magnesium layered hydroxide 3-(4-methoxyphenyl)propionic acid nanocomposite was proposed for the determination of uric acid and bisphenol A by square wave voltammetry. The results obtained denote that MLH-MPP nanocomposites enhance the sensitivity of the voltammetry detection responses. The best performance is shown by the modified carbon nanotube paste electrode (CNTPE) with the composition of single-walled carbon nanotube: magnesium layered hydroxide 3-(4-methoxyphenyl)propionic acid nanocomposite at 100:15 (% w/w). The linear range where the sensor works well is within the concentration 1.0 10-7 – 1.0 10-4 and 3.0 10-7 – 1.0 10-4 for uric acid and bisphenol A respectively with the limit of detection of 1.0 10-7 M for both organics. The interferences of uric acid and bisphenol A with other organic were studied and most of them did not interfere. The results shown for each experimental parameter on the proposed CNTPE showed that it has high sensitivity, good selectivity, repeatability and reproducibility. Therefore, the modified CNTPE can be used for the determination of uric acid and bisphenol A in real samples such as blood, plastic bottles and foods.

Keywords: bisphenol A, magnesium layered hydroxide 3-(4-methoxyphenyl)propionic acid nanocomposite, Nanocomposite, uric acid

Procedia PDF Downloads 196

Authors: Dong Hwi Kim, Dharaneedharan Subramanian, So Hyun Park, Ha-Ri Choi, Ji-Hyung Kim, Dong-Hoon Lee, Moon Soo Heo

Abstract:

The present study was performed to evaluate the use of Bacillus amyloliquefaciens JFP-2 isolated from a traditional fermented sea food, as probiotic bacteria in the diets for Rock-bream, Oplegnathus faciatus. A total of 180 fish (187.4 ± 2.7 g) were divided into two groups, control (C) and probiotic (P) group (90 fish per group) in triplicate. C group was fed with basal diet without probiotic, while P group was fed with B. amyloliquefaciens spores at concentration of 1.4 x 106 colony forming units per gram (CFU/g) of feed. After two months of feeding experiments, P group fish showed significant improvements in body weight (BW), weight gain (WG), specific growth rate (SGR) and food conversion ratio (FCR) compared with C group. Also, bi-weekly assessment of serum protein, glucose, fatty acid profile showed a significant increase in probiotic fed fish than that of control fish group. Similar increase in serum antioxidant and lysozyme activity was found in probiotic fed fish group. Twenty days challenge experiment shows decrease mortality in probiotic fed fish group when compared with that of control group. Hence, these results indicate that the use of B. amyloliquefaciens JFP-2 as a feed supplement, is beneficial to improve the health status of Oplegnathus fasciatus challenged with Vibrio anguillarum.

Keywords: Bacillus amyloliquefaciens, Oplegnathus fasciatus, probiotic feed, rock bream

Procedia PDF Downloads 242

Authors: K. Żak, S. Przetocka, R. Kitel, K. Guzik, B. Musielak, S. Malicki, G. Dubin, T. A. Holak

Abstract:

Studies on tumor genesis revealed a number of factors that may potentially serve as molecular targets for immunotherapies. One of such promising targets are PD1 and PDL1 proteins. PD1 (Programmed cell death protein 1) is expressed by activated T cells and plays a critical role in modulation of the host's immune response. One of the PD1 ligands -PDL1- is expressed by macrophages, monocytes and cancer cells which exploit it to avoid immune attack. The notion of the mechanisms used by cancer cells to block the immune system response was utilized in the development of therapies blocking PD1-PDL1 interaction. Up to date, human PD1-PDL1 complex has not been crystallized and structure of the mouse-human complex does not provide a complete view of the molecular basis of PD1-PDL1 interactions. The purpose of this study is to obtain crystal structure of the human PD1-PDL1 complex which shall allow rational design of small molecule inhibitors of the interaction. In addition, the study presents results of binding small-molecules to PD1 and fragment docking towards PD1 protein which will facilitate the design and development of small–molecule inhibitors of PD1-PDL1 interaction.

Keywords: PD1, PDL1, cancer, small molecule, drug discovery

Procedia PDF Downloads 381

Authors: Marijana M. Ačanski, Kristian A. Pastor, Djura N. Vujić

Abstract:

The identification of lipid and soluble sugar components in flour samples of different cultivars belonging to common oat species (Avena sativa L.) was performed: spring oat, winter oat and hulless oat. Fatty acids were extracted from flour samples with n-hexane, and derivatized into volatile methyl esters, using TMSH (trimethylsulfonium hydroxide in methanol). Soluble sugars were then extracted from defatted and dried samples of oat flour with 96% ethanol, and further derivatized into corresponding TMS-oximes, using hydroxylamine hydrochloride solution and BSTFA (N,O-bis-(trimethylsilyl)-trifluoroacetamide). The hexane and ethanol extracts of each oat cultivar were analyzed using GC-MS system. Lipid and simple sugar compositions are very similar in all samples of investigated cultivars. Chemometric tool was applied to numeric values of automatically integrated surface areas of detected lipid and simple sugar components in their corresponding derivatized forms. Hierarchical cluster analysis shows a very high similarity between the investigated flour samples of oat cultivars, according to the fatty acid content (0.9955). Moderate similarity was observed according to the content of soluble sugars (0.50). These preliminary results support the idea of establishing methods for oat flour authentication, and provide the means for distinguishing oat flour samples, regardless of the variety, from flour samples made of other cereal species, just by lipid and simple sugar profile analysis.

Keywords: oat cultivars, lipid composition, soluble sugar composition, GC-MS, chemometrics, authentication

Procedia PDF Downloads 284

Authors: Tian Yangyang, Xu Zihe, Lu Junping, Yang Jizhou, Xu Yiqun, Wang Jiansong, Chen Chao, Yang Mengmeng, Guo Jianhua, Mu Wenjun, Wang Guiyao, Xue Chaoqun, Liang Taibo, Hu Liwei

Abstract:

Cherry-red tobacco is receiving constant attention from cigarette enterprises because of its special flavor. This study aims to explore the material basis for the formation of the characteristic flavor of cherry-red tobacco and to clarify the distribution characteristics of the acidic aroma component groups in its mainstream smoke. In order to reach the aims of current study, this study employs GC/MS to examine the differences of distribution characteristics in particulate matter of mainstream cigarette smoke between cherry-red and common tobacco, meanwhile the aroma activity values (OVA) was used to compare the contribution of acidic aroma of cherry-red tobacco. The results showed that: 1) Isovaleric acid, acetic acid and butyric acid were the key acidic components in the mainstream smoke of the samples, followed by 3-methylvaleric acid, 4-methylvaleric acid and n-valeric acid. 2)Analysis of the release of these key sour fragrance components showed that the acidic aroma of "YUN 85" mainstream smoke was stronger than the leaf group, cherry-red tobacco was the weakest. In addition, aging had the effect of reducing the acidic components of cherry-red tobacco and the addition of cherry-red tobacco had little effect on the acidic components of the original leaf group. 3) For 14 acidic aroma(OAV>1) in smoke of cherry-red tobacco, 3-methylpentanoic acid, 4-methylpentanoic acid, pentanoic acid, and isovaleric acid were very prominent in contributing to acidic aroma, while pyruvic acid, 2-methylbutyric acid, hydrogenated acid, and propionic acid were less contribution.

Keywords: cherry-red tobacco, acidic aroma, GC/MS, mainstream cigarette smoke, odor activity value

Procedia PDF Downloads 116

Authors: Arifur Rahman, Ardeshir Amirkhani, Honghua Hu, Mark Molloy, Karen Vickery

Abstract:

Introduction and Objectives: Staphylococcus aureus and coagulase-negative staphylococci comprises approximately 65% of infections associated with medical devices and are well known for their biofilm formatting ability. Biofilm-related infections are extremely difficult to eradicate owing to their high tolerance to antibiotics and host immune defences. Currently, there is no efficient method for early biofilm detection. A better understanding to enable detection of biofilm specific proteins in vitro and in vivo can be achieved by studying planktonic and different growth phases of biofilms using a proteome analysis approach. Our goal was to construct a reference map of planktonic and biofilm associated proteins of S. aureus. Methods: S. aureus reference strain (ATCC 25923) was used to grow 24 hours planktonic, 3-day wet biofilm (3DWB), and 12-day wet biofilm (12DWB). Bacteria were grown in tryptic soy broth (TSB) liquid medium. Planktonic growth was used late logarithmic bacteria, and the Centres for Disease Control (CDC) biofilm reactor was used to grow 3 days, and 12-day hydrated biofilms, respectively. Samples were subjected to reduction, alkylation and digestion steps prior to Multiplex labelling using Tandem Mass Tag (TMT) 10-plex reagent (Thermo Fisher Scientific). The labelled samples were pooled and fractionated by high pH RP-HPLC which followed by loading of the fractions on a nanoflow UPLC system (Eksigent UPLC system, AB SCIEX). Mass spectrometry (MS) data were collected on an Orbitrap Elite (Thermo Fisher Scientific) Mass Spectrometer. Protein identification and relative quantitation of protein levels were performed using Proteome Discoverer (version 1.3, Thermo Fisher Scientific). After the extraction of protein ratios with Proteome Discoverer, additional processing, and statistical analysis was done using the TMTPrePro R package. Results and Discussion: The present study showed that a considerable proteomic difference exists among planktonic and biofilms from S. aureus. We identified 1636 total extracellular secreted proteins, of which 350 and 137 proteins of 3DWB and 12DWB showed significant abundance variation from planktonic preparation, respectively. Of these, simultaneous up-regulation in between 3DWB and 12DWB proteins such as extracellular matrix-binding protein ebh, enolase, transketolase, triosephosphate isomerase, chaperonin, peptidase, pyruvate kinase, hydrolase, aminotransferase, ribosomal protein, acetyl-CoA acetyltransferase, DNA gyrase subunit A, glycine glycyltransferase and others we found in this biofilm producer. On the contrary, simultaneous down-regulation in between 3DWB and 12DWB proteins such as alpha and delta-hemolysin, lipoteichoic acid synthase, enterotoxin I, serine protease, lipase, clumping factor B, regulatory protein Spx, phosphoglucomutase, and others also we found in this biofilm producer. In addition, we also identified a big percentage of hypothetical proteins including unique proteins. Therefore, a comprehensive knowledge of planktonic and biofilm associated proteins identified by S. aureus will provide a basis for future studies on the development of vaccines and diagnostic biomarkers. Conclusions: In this study, we constructed an initial reference map of planktonic and various growth phase of biofilm associated proteins which might be helpful to diagnose biofilm associated infections.

Keywords: bacterial biofilms, CDC bioreactor, S. aureus, mass spectrometry, TMT

Procedia PDF Downloads 155

Authors: Naruemon Prapasuwannakul

Abstract:

Soy milk residue is obtained as a byproduct from soy milk and tofu production with little economic value. It contains high protein and fiber as well as various minerals and phyto-chemical compounds. The objective of this research was to substitute soy milk residue for wheat flour in gyoza skin in order to enhance value of soy milk residue and increase protein and fiber content of gyoza skin. Wheat flour was replaced with soy milk residue from 0 to 40%. The soy milk residue prepared in this research contains 26.92% protein, 3.58% fiber, 2.88% lipid, 6.29% ash and 60.33% carbohydrate. The results showed that increasing soy milk residue decreased lightness (L*value), tensile strength and sensory attributes but increased redness (a*), yellowness (b*), protein and fiber contents of product. The result also showed that the gyoza skin substituted with 30% soy milk residue was the most acceptable (p≤0.05) and its protein and fiber content increased up to 45 % and 867 % respectively.

Keywords: Gyoza skin, sensory, soymilk residue, wheat flour

Procedia PDF Downloads 381

Authors: A. Gilewska, J. Masternak, K. Kazimierczuk, L. Turlej, J. Wietrzyk, B. Barszcz

Abstract:

Platinum-based drugs are now widely used as chemotherapeutic agents. However the platinum complexes show the toxic side-effects: i) the development of platinum resistance; ii) the occurrence of severe side effects, such as nephro-, neuro- and ototoxicity; iii) the high toxicity towards human fibroblast. Therefore the development of new anticancer drugs containing different transition-metal ions, for example, ruthenium, rhodium, iridium is a valid strategy in cancer treatment. In this paper, we reported the synthesis, spectroscopic, structural and biological properties of complexes of ruthenium, rhodium, and iridium containing N,N-chelating ligand (2,2’-bisimidazole). These complexes were characterized by elemental analysis, UV-Vis and IR spectroscopy, X-ray diffraction analysis. These complexes exhibit a typical pseudotetrahedral three-legged piano-stool geometry, in which the aromatic arene ring forms the seat of the piano-stool, while the bidentate 2,2’-bisimidazole (ligand) and the one chlorido ligand form the three legs of the stool. The spectroscopy data (IR, UV-Vis) and elemental analysis correlate very well with molecular structures. Moreover, the cytotoxic activity of the complexes was carried out on human cancer cell lines: LoVo (colorectal adenoma), MV-4-11 (myelomonocytic leukaemia), MCF-7 (breast adenocarcinoma) and normal healthy mouse fibroblast BALB/3T3 cell lines. To predict a binding mode, a potential interaction of metal complexes with calf thymus DNA (CT-DNA) and protein (BSA) has been explored using UV absorption and circular dichroism (CD). It is interesting to note that the investigated complexes show no cytotoxic effect towards the normal BALB/3T3 cell line, compared to cisplatin, which IC₅₀ values was determined as 2.20 µM. Importantly, Ru(II) displayed the highest activity against HL-60 (IC₅₀ 4.35 µM). The biological studies (UV-Vis and circular dichroism) suggest that arene-complexes could interact with calf thymus DNA probably via an outside binding mode and interact with protein (BSA).

Keywords: ruthenium(II) complex, rhodium(III) complex, iridium(III) complex, biological activity

Procedia PDF Downloads 117

Authors: Chowon Kim, Yoo-Kyung Kim, Kyeong Yeon Park, Hyun-Shik Lee

Abstract:

Perfluoroalkyl compounds (PFCs) are globally distributed synthetic compounds that are known to adversely affect human health. Developmental toxicity assessment of PFCs is important to facilitate the evaluation of their environmental impact. In the present study, we assessed the developmental toxicity and teratogenicity of PFCs with different numbers of carbon atoms on Xenopus embryogenesis. An initial frog embryo teratogenicity assay-Xenopus (FETAX) assay was performed that identified perfluorohexanoic (PFHxA) and perfluoroheptanoic (PFHpA) acids as potential teratogens and developmental toxicants. The mechanism underlying this teratogenicity was also investigated by measuring the expression of tissue-specific biomarkers such as phosphotyrosine‑binding protein, xPTB (liver); NKX2.5 (heart); and Cyl18 (intestine). Whole‑mount in situ hybridization, reverse transcriptase‑polymerase chain reaction (RT-PCR), and histologic analyses detected severe defects in the liver and heart following exposure to PFHxA or PFHpA. In addition, immunoblotting revealed that PFHpA significantly increased the phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), while PFHxA slightly increased these, as compared with the control. These results suggest that PFHxA and PFHpA are developmental toxicants and teratogens, with PFHpA producing more severe effects on liver and heart development through the induction of ERK and JNK phosphorylation.

Keywords: PFCs, ERK, JNK, xenopus

Procedia PDF Downloads 282

Authors: Uzma Saqib, Mirza S. Baig

Abstract:

Myeloid differentiation primary response protein 88 (MYD88) has long been considered a central player in the inflammatory pathway. Recent studies clearly suggest that it is an important therapeutic target in inflammation. On the other hand, a recent study on the interaction between the orphan nuclear receptor (Nur77) and p38α, leading to increased lipopolysaccharide-induced hyperinflammatory response, suggests this binary complex as a therapeutic target. In this study, we have designed inhibitors that can inhibit both MYD88 and Nur77 at the same time. Since both MYD88 and Nur77 are an integral part of the pathways involving lipopolysaccharide-induced activation of NF-κB-mediated inflammation, we tried to target both proteins with the same library in order to retrieve compounds having dual inhibitory properties. To perform this, we developed a homodimeric model of MYD88 and, along with the crystal structure of Nur77, screened a virtual library of compounds from the traditional Chinese medicine database containing ~61,000 compounds. We analyzed the resulting hits for their efficacy for dual binding and probed them for developing a common pharmacophore model that could be used as a prototype to screen compound libraries as well as to guide combinatorial library design to search for ideal dual-target inhibitors. Thus, our study explores the identification of novel leads having dual inhibiting effects due to binding to both MYD88 and Nur77 targets.

Keywords: drug design, Nur77, MYD88, inflammation

Procedia PDF Downloads 287