Search results for: physicochemical properties of amino acid
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 11597

Search results for: physicochemical properties of amino acid

10937 Physicochemical Characterizations of Marine and River Sediments in the North of France

Authors: Abriak Nor Edine, Zentar Rachid, Achour Raouf, Tran Ngoc Thanh

Abstract:

This work is undertaken to develop a methodology to enhance the management of dredged marine and river sediments in the North of France. The main objective of this study is to determine the main characteristics of these sediments. In this order, physical, mineralogical and chemical properties of both types of sediments are measured. Moreover, their potential impacts on the environment are assessed throughout leaching tests. From the obtained results, the potential of their use in road engineering is discussed.

Keywords: marine sediments, river sediments, physico chemical characterizations, environmental characterizations

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10936 A Kunitz-Type Serine Protease Inhibitor from Rock Bream, Oplegnathus fasciatus Involved in Immune Responses

Authors: S. D. N. K. Bathige, G. I. Godahewa, Navaneethaiyer Umasuthan, Jehee Lee

Abstract:

Kunitz-type serine protease inhibitors (KTIs) are identified in various organisms including animals, plants and microbes. These proteins shared single or multiple Kunitz inhibitory domains link together or associated with other types of domains. Characteristic Kunitz type domain composed of around 60 amino acid residues with six conserved cysteine residues to stabilize by three disulfide bridges. KTIs are involved in various physiological processes, such as ion channel blocking, blood coagulation, fibrinolysis and inflammation. In this study, two Kunitz-type domain containing protein was identified from rock bream database and designated as RbKunitz. The coding sequence of RbKunitz encoded for 507 amino acids with 56.2 kDa theoretical molecular mass and 5.7 isoelectric point (pI). There are several functional domains including MANEC superfamily domain, PKD superfamily domain, and LDLa domain were predicted in addition to the two characteristic Kunitz domain. Moreover, trypsin interaction sites were also identified in Kunitz domain. Homology analysis revealed that RbKunitz shared highest identity (77.6%) with Takifugu rubripes. Completely conserved 28 cysteine residues were recognized, when comparison of RbKunitz with other orthologs from different taxonomical groups. These structural evidences indicate the rigidity of RbKunitz folding structure to achieve the proper function. The phylogenetic tree was constructed using neighbor-joining method and exhibited that the KTIs from fish and non-fish has been evolved in separately. Rock bream was clustered with Takifugu rubripes. The SYBR Green qPCR was performed to quantify the RbKunitz transcripts in different tissues and challenged tissues. The mRNA transcripts of RbKunitz were detected in all tissues (muscle, spleen, head kidney, blood, heart, skin, liver, intestine, kidney and gills) analyzed and highest transcripts level was detected in gill tissues. Temporal transcription profile of RbKunitz in rock bream blood tissues was analyzed upon LPS (lipopolysaccharide), Poly I:C (Polyinosinic:polycytidylic acid) and Edwardsiella tarda challenge to understand the immune responses of this gene. Compare to the unchallenged control RbKunitz exhibited strong up-regulation at 24 h post injection (p.i.) after LPS and E. tarda injection. Comparatively robust expression of RbKunits was observed at 3 h p.i. upon Poly I:C challenge. Taken together all these data indicate that RbKunitz may involve into to immune responses upon pathogenic stress, in order to protect the rock bream.

Keywords: Kunitz-type, rock bream, immune response, serine protease inhibitor

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10935 Isolation and Characterization of Indigenous Rhizosphere Bacteria Producing Gibberellin Acid from Local Soybeans in Three Different Areas of South Sulawesi

Authors: Asmiaty Sahur, Ambo Ala, Baharuddin Patanjengi, Elkawakib Syam'un

Abstract:

This study aimed to isolate and characterize the indigenous Rhizosphere bacteria producing Gibberellin Acid as plant growth isolated from local soybean of three different areas in South Sulawesi, Indonesia. Several soil samples of soybean plants were collected from the Rhizosphere of local soybeans in three different areas of South Sulawesi such as Soppeng, Bone and Takalar. There were 56 isolates of bacteria were isolated and grouped into gram-positive bacteria and gram negative bacteria .There are 35 isolates produce a thick slime or slimy when cultured on media Natrium Broth and the remaining of those produced spores. The results showed that of potential bacterial isolated produced Gibberellin Acid in high concentration. The best isolate of Rhizosphere bacteria for the production of Gibberellin Acid is with concentration 2%. There are 4 isolates that had higher concentration are AKB 19 (4.67 mg/ml) followed by RKS 17 (3.80 mg/ml), RKS 25 (3.70 mg / ml) and RKS 24 (3.29 mg/ml) respectively.

Keywords: rhizosphere, bacteria, gibberellin acid, soybeans

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10934 Impact of Sericin Treatment on Perfection Dyeing of Polyester Viscose Blend

Authors: Omaima G. Allam, O. A. Hakeim, K. Haggag, N. S. Elshemy

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In the midst of the two decades the use of microwave dielectric warming in the field of science has transformed into a powerful methodology to redesign compound procedures. The potential benefit of the application of these modern methods of treatment emphasize so as to reach to optimum treatment conditions and the best results, especially hydrophobicity, moisture content and increase dyeing processing while maintaining the physical and chemical properties of each textile. Moreover, polyester fibres are sometimes spun together with natural fibres to produce a cloth with blended properties. So that at the present task, the polyester/viscose mix fabrics (60 /40) were pretreated with 4 g/l of KOH for 2 min in microwave irradiation with a liquor ratio 1:25. Subsequently fabrics were inundated with different concentrations of sericin (10, 30, 50 g/l). Treated fabrics were coloured with the commercial dyes samples: Reactive Red 84(Dye 1). C. I. Acid Blue 203(Dye 2) and C.I. Reactive violet 5 (Dye 3). Colour value was specified as well as fastness properties. Likewise, the physical properties of untreated and treated fabrics such as moisture content %, tensile strength, elongation % and were evaluated. The untreated and treated fabrics are described by infrared spectroscopy (FTIR) and scanning electron microscopy.

Keywords: polyester viscose blends fabric, sericin, dyes, colour value

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10933 Simultaneous Production of Forskolin and Rosmarinic Acid in vitro Cultures of Coleus Forskohlii Briq

Authors: Ennus Tajuddin Tamboli, Madhukar Garg, Mohd. Mujeeb, Sayeed Ahmad

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An efficient protocol for simultaneous production of forskolin and rosmarinic acid in in vitro callus derived from the leaves of Coleus forskohlii Briq. has been developed. MS media was used for the establishment of cultures and NAA + 6-BA (1.0 ppm) was found best for callus growth. The callus was further subjected to treatment with various elicitor/precursors viz. chitosan, thidiazuron and methyl jasmonate to observe their effect on production of biomass and accumulation of secondary metabolites. The content of forskolin and rosmarinic acid were estimated by HPTLC, in comparison to natural explant which showed 2 fold and 10 fold rise in forskolin and rosmarinic acid content, respectively. Methy1 jasmonate 50 µM was found best for production of forskolin, whereas thidiazuron showed best results in the yield of rosmarinic acid, separately in static culture. However, combined treatment in suspension culture showed moderated effect for increase in secondary metabolites but the biomass increased significantly as compared to static culture.

Keywords: plant tissue culture, secondary metabolites, coleus, forskolin, rosmarinic acid, HPTLC

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10932 Incorporation of Noncanonical Amino Acids into Hard-to-Express Antibody Fragments: Expression and Characterization

Authors: Hana Hanaee-Ahvaz, Monika Cserjan-Puschmann, Christopher Tauer, Gerald Striedner

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Incorporation of noncanonical amino acids (ncAA) into proteins has become an interesting topic as proteins featured with ncAAs offer a wide range of different applications. Nowadays, technologies and systems exist that allow for the site-specific introduction of ncAAs in vivo, but the efficient production of proteins modified this way is still a big challenge. This is especially true for 'hard-to-express' proteins where low yields are encountered even with the native sequence. In this study, site-specific incorporation of azido-ethoxy-carbonyl-Lysin (azk) into an anti-tumor-necrosis-factor-α-Fab (FTN2) was investigated. According to well-established parameters, possible site positions for ncAA incorporation were determined, and corresponding FTN2 genes were constructed. Each of the modified FTN2 variants has one amber codon for azk incorporated either in its heavy or light chain. The expression level for all variants produced was determined by ELISA, and all azk variants could be produced with a satisfactory yield in the range of 50-70% of the original FTN2 variant. In terms of expression yield, neither the azk incorporation position nor the subunit modified (heavy or light chain) had a significant effect. We confirmed correct protein processing and azk incorporation by mass spectrometry analysis, and antigen-antibody interaction was determined by surface plasmon resonance analysis. The next step is to characterize the effect of azk incorporation on protein stability and aggregation tendency via differential scanning calorimetry and light scattering, respectively. In summary, the incorporation of ncAA into our Fab candidate FTN2 worked better than expected. The quantities produced allowed a detailed characterization of the variants in terms of their properties, and we can now turn our attention to potential applications. By using click chemistry, we can equip the Fabs with additional functionalities and make them suitable for a wide range of applications. We will now use this option in a first approach and develop an assay that will allow us to follow the degradation of the recombinant target protein in vivo. Special focus will be laid on the proteolytic activity in the periplasm and how it is influenced by cultivation/induction conditions.

Keywords: degradation, FTN2, hard-to-express protein, non-canonical amino acids

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10931 Evaluation of Functional Properties of Protein Hydrolysate from the Fresh Water Mussel Lamellidens marginalis for Nutraceutical Therapy

Authors: Jana Chakrabarti, Madhushrita Das, Ankhi Haldar, Roshni Chatterjee, Tanmoy Dey, Pubali Dhar

Abstract:

High incidences of Protein Energy Malnutrition as a consequence of low protein intake are quite prevalent among the children in developing countries. Thus prevention of under-nutrition has emerged as a critical challenge to India’s developmental Planners in recent times. Increase in population over the last decade has led to greater pressure on the existing animal protein sources. But these resources are currently declining due to persistent drought, diseases, natural disasters, high-cost of feed, and low productivity of local breeds and this decline in productivity is most evident in some developing countries. So the need of the hour is to search for efficient utilization of unconventional low-cost animal protein resources. Molluscs, as a group is regarded as under-exploited source of health-benefit molecules. Bivalve is the second largest class of phylum Mollusca. Annual harvests of bivalves for human consumption represent about 5% by weight of the total world harvest of aquatic resources. The freshwater mussel Lamellidens marginalis is widely distributed in ponds and large bodies of perennial waters in the Indian sub-continent and well accepted as food all over India. Moreover, ethno-medicinal uses of the flesh of Lamellidens among the rural people to treat hypertension have been documented. Present investigation thus attempts to evaluate the potential of Lamellidens marginalis as functional food. Mussels were collected from freshwater ponds and brought to the laboratory two days before experimentation for acclimatization in laboratory conditions. Shells were removed and fleshes were preserved at- 20oC until analysis. Tissue homogenate was prepared for proximate studies. Fatty acids and amino acids composition were analyzed. Vitamins, Minerals and Heavy metal contents were also studied. Mussel Protein hydrolysate was prepared using Alcalase 2.4 L and degree of hydrolysis was evaluated to analyze its Functional properties. Ferric Reducing Antioxidant Power (FRAP) and DPPH Antioxidant assays were performed. Anti-hypertensive property was evaluated by measuring Angiotensin Converting Enzyme (ACE) inhibition assay. Proximate analysis indicates that mussel meat contains moderate amount of protein (8.30±0.67%), carbohydrate (8.01±0.38%) and reducing sugar (4.75±0.07%), but less amount of fat (1.02±0.20%). Moisture content is quite high but ash content is very low. Phospholipid content is significantly high (19.43 %). Lipid constitutes, substantial amount of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) which have proven prophylactic values. Trace elements are found present in substantial amount. Comparative study of proximate nutrients between Labeo rohita, Lamellidens and cow’s milk indicates that mussel meat can be used as complementary food source. Functionality analyses of protein hydrolysate show increase in Fat absorption, Emulsification, Foaming capacity and Protein solubility. Progressive anti-oxidant and anti-hypertensive properties have also been documented. Lamellidens marginalis can thus be regarded as a functional food source as this may combine effectively with other food components for providing essential elements to the body. Moreover, mussel protein hydrolysate provides opportunities for utilizing it in various food formulations and pharmaceuticals. The observations presented herein should be viewed as a prelude to what future holds.

Keywords: functional food, functional properties, Lamellidens marginalis, protein hydrolysate

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10930 Control of Staphylococcus aureus in Meat System by in situ and ex situ Bacteriocins from Lactobacillus sakei and Pediococcus spp.

Authors: M. Naimi, M. B. Khaled

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The present study consisted of an applied test in meat system to assess the effectiveness of three bio agents bacteriocinproducing strains: Lm24: Lactobacillus sakei, Lm14and Lm25: Pediococcus spp. Two tests were carried out: The ex-situ test was intended for three batches added with crude bacteriocin solutions at 12.48 AU/ml for Lm25 and 8.4 AU/ml for Lm14 and Lm24. However, the in situ one consisted of four batches; three of them inoculated with one bacteriocinogenic Lm25, Lm14, Lm24, respectively. The fourth one was used in mixture: Lm14+m24 at approximately of 107 CFU/ml. The two used tests were done in the presence of the pathogen St. aureus ATCC 6538, as a test strain at 103 CFU/ml. Another batch served as a positive or a negative control was used too. The incubation was performed at 7°C. Total viable counts, staphylococci and lactic acid bacteria, at the beginning and at selected times with interval of three days were enumerated. Physicochemical determinations (except for in situ test): pH, dry mater, sugars, fat and total protein, at the beginning and at end of the experiment, were done, according to the international norms. Our results confirmed the ex situ effectiveness. Furthermore, the batches affected negatively the total microbial load over the incubation days, and showed a significant regression in staphylococcal load at day seven, for Lm14, Lm24, and Lm25 of 0.73, 2.11, and 2.4 log units. It should be noticed that, at the last day of culture, staphylococcal load was nil for the three batches. In the in situ test, the cultures displayed less inhibitory attitude and recorded a decrease in staphylococcal load, for Lm14, Lm24, Lm25, Lm14+m24 of 0.73, 0.20, 0.86, 0.032 log units. Therefore, physicochemical analysis for Lm14, Lm24, Lm25, Lm14+m24 showed an increase in pH from 5.50 to 5.77, 6.18, 5.96, 7.22, a decrease in dry mater from 7.30% to 7.05%, 6.87%, 6.32%, 6.00%.This result reflects the decrease in fat ranging from 1.53% to 1.49%, 1.07%, 0.99%, 0.87%; and total protein from 6.18% to 5.25%, 5.56%, 5.37%, 5.5%. This study suggests that the use of selected strains as Lm25 could lead to the best results and would help in preserving and extending the shelf life of lamb meat.

Keywords: biocontrol, in situ, ex situ, meat system, St. aureus, Lactobacillus sakei, Pediococcus spp.

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10929 Using Hemicellulosic Liquor from Sugarcane Bagasse to Produce Second Generation Lactic Acid

Authors: Regiane A. Oliveira, Carlos E. Vaz Rossell, Rubens Maciel Filho

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Lactic acid, besides a valuable chemical may be considered a platform for other chemicals. In fact, the feasibility of hemicellulosic sugars as feedstock for lactic acid production process, may represent the drop of some of the barriers for the second generation bioproducts, especially bearing in mind the 5-carbon sugars from the pre-treatment of sugarcane bagasse. Bearing this in mind, the purpose of this study was to use the hemicellulosic liquor from sugarcane bagasse as a substrate to produce lactic acid by fermentation. To release of sugars from hemicellulose it was made a pre-treatment with a diluted sulfuric acid in order to obtain a xylose's rich liquor with low concentration of inhibiting compounds for fermentation (≈ 67% of xylose, ≈ 21% of glucose, ≈ 10% of cellobiose and arabinose, and around 1% of inhibiting compounds as furfural, hydroxymethilfurfural and acetic acid). The hemicellulosic sugars associated with 20 g/L of yeast extract were used in a fermentation process with Lactobacillus plantarum to produce lactic acid. The fermentation process pH was controlled with automatic injection of Ca(OH)2 to keep pH at 6.00. The lactic acid concentration remained stable from the time when the glucose was depleted (48 hours of fermentation), with no further production. While lactic acid is produced occurs the concomitant consumption of xylose and glucose. The yield of fermentation was 0.933 g lactic acid /g sugars. Besides, it was not detected the presence of by-products, what allows considering that the microorganism uses a homolactic fermentation to produce its own energy using pentose-phosphate pathway. Through facultative heterofermentative metabolism the bacteria consume pentose, as is the case of L. plantarum, but the energy efficiency for the cell is lower than during the hexose consumption. This implies both in a slower cell growth, as in a reduction in lactic acid productivity compared with the use of hexose. Also, L. plantarum had shown to have a capacity for lactic acid production from hemicellulosic hydrolysate without detoxification, which is very attractive in terms of robustness for an industrial process. Xylose from hydrolyzed bagasse and without detoxification is consumed, although the hydrolyzed bagasse inhibitors (especially aromatic inhibitors) affect productivity and yield of lactic acid. The use of sugars and the lack of need for detoxification of the C5 liquor from sugarcane bagasse hydrolyzed is a crucial factor for the economic viability of second generation processes. Taking this information into account, the production of second generation lactic acid using sugars from hemicellulose appears to be a good alternative to the complete utilization of sugarcane plant, directing molasses and cellulosic carbohydrates to produce 2G-ethanol, and hemicellulosic carbohydrates to produce 2G-lactic acid.

Keywords: fermentation, lactic acid, hemicellulosic sugars, sugarcane

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10928 A Similarity/Dissimilarity Measure to Biological Sequence Alignment

Authors: Muhammad A. Khan, Waseem Shahzad

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Analysis of protein sequences is carried out for the purpose to discover their structural and ancestry relationship. Sequence similarity determines similar protein structures, similar function, and homology detection. Biological sequences composed of amino acid residues or nucleotides provide significant information through sequence alignment. In this paper, we present a new similarity/dissimilarity measure to sequence alignment based on the primary structure of a protein. The approach finds the distance between the two given sequences using the novel sequence alignment algorithm and a mathematical model. The algorithm runs at a time complexity of O(n²). A distance matrix is generated to construct a phylogenetic tree of different species. The new similarity/dissimilarity measure outperforms other existing methods.

Keywords: alignment, distance, homology, mathematical model, phylogenetic tree

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10927 Diamond-Like Carbon-Based Structures as Functional Layers on Shape-Memory Alloy for Orthopedic Applications

Authors: Piotr Jablonski, Krzysztof Mars, Wiktor Niemiec, Agnieszka Kyziol, Marek Hebda, Halina Krawiec, Karol Kyziol

Abstract:

NiTi alloys, possessing unique mechanical properties such as pseudoelasticity and shape memory effect (SME), are suitable for many applications, including implanthology and biomedical devices. Additionally, these alloys have similar values of elastic modulus to those of human bones, what is very important in orthopedics. Unfortunately, the environment of physiological fluids in vivo causes unfavorable release of Ni ions, which in turn may lead to metalosis as well as allergic reactions and toxic effects in the body. For these reasons, the surface properties of NiTi alloys should be improved to increase corrosion resistance, taking into account biological properties, i.e. excellent biocompatibility. The prospective in this respect are layers based on DLC (Diamond-Like Carbon) structures, which are an attractive solution for many applications in implanthology. These coatings (DLC), usually obtained by PVD (Physical Vapour Deposition) and PA CVD (Plasma Activated Chemical Vapour Deposition) methods, can be also modified by doping with other elements like silicon, nitrogen, oxygen, fluorine, titanium and silver. These methods, in combination with a suitably designed structure of the layers, allow the possibility co-decide about physicochemical and biological properties of modified surfaces. Mentioned techniques provide specific physicochemical properties of substrates surface in a single technological process. In this work, the following types of layers based on DLC structures (incl. Si-DLC or Si/N-DLC) were proposed as prospective and attractive approach in surface functionalization of shape memory alloy. Nitinol substrates were modified in plasma conditions, using RF CVD (Radio Frequency Chemical Vapour Deposition). The influence of plasma treatment on the useful properties of modified substrates after deposition DLC layers doped with silica and/or nitrogen atoms, as well as only pre-treated in O2 NH3 plasma atmosphere in a RF reactor was determined. The microstructure and topography of the modified surfaces were characterized using scanning electron microscopy (SEM) and atomic force microscopy (AFM). Furthermore, the atomic structure of coatings was characterized by IR and Raman spectroscopy. The research also included the evaluation of surface wettability, surface energy as well as the characteristics of selected mechanical and biological properties of the layers. In addition, the corrosion properties of alloys after and before modification in the physiological saline were also investigated. In order to determine the corrosion resistance of NiTi in the Ringer solution, the potentiodynamic polarization curves (LSV – Linear Sweep Voltamperometry) were plotted. Furthermore, the evolution of corrosion potential versus immersion time of TiNi alloy in Ringer solution was performed. Based on all carried out research, the usefullness of proposed modifications of nitinol for medical applications was assessed. It was shown, inter alia, that the obtained Si-DLC layers on the surface of NiTi alloy exhibit a characteristic complex microstructure, increased surface development, which is an important aspect in improving the osteointegration of an implant. Furthermore, the modified alloy exhibits biocompatibility, the transfer of the metal (Ni, Ti) to Ringer’s solution is clearly limited.

Keywords: bioactive coatings, corrosion resistance, doped DLC structure, NiTi alloy, RF CVD

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10926 Isolation and Identification of Cytotoxic Compounds from Fruticose Lichen Roccella montagnei, and It’s in Silico Docking Study against CDK-10

Authors: Tripti Mishra, Shipra Shukla, Sanjeev Meena, , Ruchi Singh, Mahesh Pal, D. K. Upreti, Dipak Datta

Abstract:

Roccella montagnei belongs to lichen family Roccelleceae growing luxuriantly along the coastal regions of India. As Roccella has been shown to be bioactive, we prepared methanolic extract and assessed its anticancer potential. The methanolic extract showed significant in vitro cytotoxic activity against four human cancer cell lines such as Colon (DLD-1, SW-620), Breast (MCF-7), Head and Neck (FaDu). This prompted us to isolate bioactive compounds through column chromatography. Two compounds Roccellic acid and Everninic acid have been isolated, out of which Everninic acid is reported for the first time. Both the compounds have been tested for in vitro cytotoxic activity in which Roccellic acid showed strong anticancer activity as compared to the Everninic acid. CDK-10 (Cyclin-dependent kinase) contributes to proliferation of cancer cells, and aberrant activity of these kinases has been reported in a wide variety of human cancers. These kinases, therefore, constitute biomarkers of proliferation and attractive pharmacological targets for the development of anticancer therapeutics. Therefore both the isolated compounds were tested for in silico molecular docking study against CDK-10 isomer enzyme to support the cytotoxic activity.

Keywords: cytotoxic activity, everninic acid, roccellic acid, R. montagnei

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10925 iPSCs More Effectively Differentiate into Neurons on PLA Scaffolds with High Adhesive Properties for Primary Neuronal Cells

Authors: Azieva A. M., Yastremsky E. V., Kirillova D. A., Patsaev T. D., Sharikov R. V., Kamyshinsky R. A., Lukanina K. I., Sharikova N. A., Grigoriev T. E., Vasiliev A. L.

Abstract:

Adhesive properties of scaffolds, which predominantly depend on the chemical and structural features of their surface, play the most important role in tissue engineering. The basic requirements for such scaffolds are biocompatibility, biodegradation, high cell adhesion, which promotes cell proliferation and differentiation. In many cases, synthetic polymers scaffolds have proven advantageous because they are easy to shape, they are tough, and they have high tensile properties. The regeneration of nerve tissue still remains a big challenge for medicine, and neural stem cells provide promising therapeutic potential for cell replacement therapy. However, experiments with stem cells have their limitations, such as low level of cell viability and poor control of cell differentiation. Whereas the study of already differentiated neuronal cell culture obtained from newborn mouse brain is limited only to cell adhesion. The growth and implantation of neuronal culture requires proper scaffolds. Moreover, the polymer scaffolds implants with neuronal cells could demand specific morphology. To date, it has been proposed to use numerous synthetic polymers for these purposes, including polystyrene, polylactic acid (PLA), polyglycolic acid, and polylactide-glycolic acid. Tissue regeneration experiments demonstrated good biocompatibility of PLA scaffolds, despite the hydrophobic nature of the compound. Problem with poor wettability of the PLA scaffold surface could be overcome in several ways: the surface can be pre-treated by poly-D-lysine or polyethyleneimine peptides; roughness and hydrophilicity of PLA surface could be increased by plasma treatment, or PLA could be combined with natural fibers, such as collagen or chitosan. This work presents a study of adhesion of both induced pluripotent stem cells (iPSCs) and mouse primary neuronal cell culture on the polylactide scaffolds of various types: oriented and non-oriented fibrous nonwoven materials and sponges – with and without the effect of plasma treatment and composites with collagen and chitosan. To evaluate the effect of different types of PLA scaffolds on the neuronal differentiation of iPSCs, we assess the expression of NeuN in differentiated cells through immunostaining. iPSCs more effectively differentiate into neurons on PLA scaffolds with high adhesive properties for primary neuronal cells.

Keywords: PLA scaffold, neurons, neuronal differentiation, stem cells, polylactid

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10924 Polymer Nanocarrier for Rheumatoid Arthritis Therapy

Authors: Vijayakameswara Rao Neralla, Jueun Jeon, Jae Hyung Park

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To develop a potential nanocarrier for diagnosis and treatment of rheumatoid arthritis (RA), we prepared a hyaluronic acid (HA)-5β-cholanic acid (CA) conjugate with an acid-labile ketal linker. This conjugate could self-assemble in aqueous conditions to produce pH-responsive HA-CA nanoparticles as potential carriers of the anti-inflammatory drug methotrexate (MTX). MTX was rapidly released from nanoparticles under inflamed synovial tissue in RA. In vitro cytotoxicity data showed that pH-responsive HA-CA nanoparticles were non-toxic to RAW 264.7 cells. In vivo biodistribution results confirmed that, after their systemic administration, pH-responsive HA-CA nanoparticles selectively accumulated in the inflamed joints of collagen-induced arthritis mice. These results indicate that pH-responsive HA-CA nanoparticles represent a promising candidate as a drug carrier for RA therapy.

Keywords: rheumatoid arthritis, hyaluronic acid, nanocarrier, self-assembly, MTX

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10923 Static Characterization of a Bio-Based Sandwich in a Humid Environment

Authors: Zeineb Kesentini, Abderrahim El Mahi, Jean Luc Rebiere, Rachid El Guerjouma, Moez Beyaoui, Mohamed Haddar

Abstract:

Industries’ attention has been drawn to green and sustainable materials as a result of the present energy deficit and environmental damage. Sandwiches formed of auxetic structures made up of periodic cells are also being investigated by industry. Several tests have emphasized the exceptional properties of these materials. In this study, the sandwich's core is a one-cell auxetic core. Among plant fibers, flax fibers are chosen because of their good mechanical properties comparable to those of glass fibers. Poly (lactic acid) (PLA), as a green material, is available from starch, and its production process requires fewer fossil resources than petroleum-based plastics. A polylactic acid (PLA) reinforced with flax fiber filament was employed in this study. The manufacturing process used to manufacture the test specimens is 3D printing. The major drawback of a 100% bio-based material is its low resistance to moisture absorption. In this study, a sandwich based on PLA / flax with an auxetic core is characterized statically for different periods of immersion in water. Bending tests are carried out on the composite sandwich for three immersion time. Results are compared to those of non immersed specimens. It is found that non aged sandwich has the ultimate bending stiffness.

Keywords: auxetic, bending tests, biobased composite, sandwich structure, 3D printing

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10922 Application of Flory Paterson’s Theory on the Volumetric Properties of Liquid Mixtures: 1,2-Dichloroethane with Aliphatic and Cyclic Ethers

Authors: Linda Boussaid, Farid Brahim Belaribi

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The physico-chemical properties of liquid materials in the industrial field, in general, and in that of the chemical industries, in particular, constitutes a prerequisite for the design of equipment, for the resolution of specific problems (related to the techniques of purification and separation, at risk in the transport of certain materials, etc.) and, therefore, at the production stage. Chloroalkanes, ethers constitute three chemical families having an industrial, theoretical and environmental interest. For example, these compounds are used in various applications in the chemical and pharmaceutical industries. In addition, they contribute to the particular thermodynamic behavior (deviation from ideality, association, etc.) of certain mixtures which constitute a severe test for predictive theoretical models. Finally, due to the degradation of the environment in the world, a renewed interest is observed for ethers, because some of their physicochemical properties could contribute to lower pollution (ethers would be used as additives in aqueous fuels.). This work is a thermodynamic, experimental and theoretical study of the volumetric properties of liquid binary systems formed from compounds belonging to the chemical families of chloroalkanes, ethers, having an industrial, theoretical and environmental interest. Experimental determination of the densities and excess volumes of the systems studied, at different temperatures in the interval [278.15-333.15] K and at atmospheric pressure, using an AntonPaar vibrating tube densitometer of the DMA5000 type. This contribution of experimental data, on the volumetric properties of the binary liquid mixtures of 1,2-dichloroethane with an ether, supplemented by an application of the theoretical model of Prigogine-Flory-Patterson PFP, will probably contribute to the enrichment of the thermodynamic database and the further development of the theory of Flory in its Prigogine-Flory-Patterson (PFP) version, for a better understanding of the thermodynamic behavior of these liquid binary mixtures

Keywords: prigogine-flory-patterson (pfp), propriétés volumétrique , volume d’excés, ethers

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10921 Dissolution of South African Limestone for Wet Flue Gas Desulphurization

Authors: Lawrence Koech, Ray Everson, Hein Neomagus, Hilary Rutto

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Wet Flue gas desulphurization (FGD) systems are commonly used to remove sulphur dioxide from flue gas by contacting it with limestone in aqueous phase which is obtained by dissolution. Dissolution is important as it affects the overall performance of a wet FGD system. In the present study, effects of pH, stirring speed, solid to liquid ratio and acid concentration on the dissolution of limestone using an organic acid (adipic acid) were investigated. This was investigated using the pH stat apparatus. Calcium ions were analyzed at the end of each experiment using Atomic Absorption (AAS) machine.

Keywords: desulphurization, limestone, dissolution, pH stat apparatus

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10920 Ag Nanoparticle/Melamine Sulfonic Acid Supported on Alumina: Efficient Catalytic System in Synthesis of Dihydropyrimidines

Authors: Parya Nasehi, Mohammad Kazem Mohammadi

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3,4-dihydropyrimidin-2(1H)-thiones were synthesized in the presence of Ag nanoparticle/melamine sulfonic acid (MSA) supported on alumina. The reaction was carried out at 110 oC for 20 min under solvent free conditions. This method have some advantages such as good yield, mild reaction conditions, ease of operation and work up, short reaction time and high product purity.

Keywords: nanoparticle melamine sulfonic acid, Al2O3, Biginelli reaction, 3, 4-dihydropyrimidin-2(1H, solvent free

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10919 Desirable Fatty Acids in Meat of Cattle Fed Different Levels of Lipid-Based Diets

Authors: Tiago N. P. Valente, Erico S. Lima, Roberto O. Roça

Abstract:

Introduction: Research has stimulated animal production studies on solutions to decrease the level of saturated fatty acids and increase unsaturated in foods of animal origin. The objective of this study was to determine the effect of the dietary inclusion of lipid-based diets on the fatty acid profiles from finishing cattle. Materials and Methods: The study was carried out in the Chapéu de Couro Farm in Aguaí/SP, Brazil. A group of 39 uncastrated Nellore cattle. Mean age of the animals was 36 months, and initial mean live weight was 494.1 ± 10.1. Animals were randomly assigned to one of three treatments, based on dry matter: feed with control diet 2.50% cottonseed, feed with 11.50% cottonseed, and feed with 3.13% cottonseed added of 1.77% protected lipid. Forage:concentrate ratio was 50:50 on a dry matter basis. Sugar cane chopped was used as forage. After 63 days mean final live weight was 577.01 kg ± 11.34. After slaughter, carcasses were identified and divided into two halves that were kept in a cold chamber for 24 hours at 2°C. Then, part of the M. longissimus thoracis of each animal was removed between the 12th and 13th rib of the left half carcass. The samples steaks were 2.5 cm thick and were identified and stored frozen in a freezer at -18°C. The analysis of methyl esters of fatty acids was carried out in a gas chromatograph. Desirable fatty acids (FADes) were determined by the sum of unsaturated fatty acids and stearic acid (C18:0). Results and Discussion: No differences (P>0.05) between the diets for the proportion of FADes in the meat of the animals in this study, according to the lipid sources used. The inclusion of protected fat or cottonseed in the diet did not change the proportion of FADes in the meat. The proportion mean of FADes in meat in the present study were: as pentadecanoic acid (C15:1 = 0.29%), palmitoleic acid (C16:1 = 4.26%), heptadecanoic acid (C17:1 = 0.07%), oleic acid (C18:1n9c = 37.32%), γ-linolenic acid (0.94%) and α-linolenic acid (1.04%), elaidic acid (C18:1n9t = 0.50%), eicosatrienoic acid (C20:3n3 = 0.03%), eicosapentaenoic acid (C20:5n3 = 0.04%), erucic acid (C22:1n9 = 0.89%), docosadienoic acid (C22:2 = 0.04%) and stearic acid (C18:0 = 21.53%). Conclusions: The add the cottonseed or protected lipid in diet is not affected fatty acids profiles the desirable fatty acids in meat. Acknowledgements: IFGoiano, FAPEG and CNPq (Brazil).

Keywords: beef quality, cottonseed, protected fat, unsaturated fatty acids

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10918 Consequence of Multi-Templating of Closely Related Structural Analogues on a Chitosan-Methacryllic Acid Molecularly Imprinted Polymer Matrix-Thermal and Chromatographic Traits

Authors: O.Ofoegbu, S. Roongnapa, A.N. Eboatu

Abstract:

Most polluted environments, most challengingly, aerosol types, contain a cocktail of different toxicants. Multi-templating of matrices have been the recent target by researchers in a bid to solving complex mixed-toxicant challenges using single or common remediation systems. This investigation looks at the effect of such multi-templated system vis-a-vis the synthesis by non-covalent interaction, of a molecularly imprinted polymer architecture using nicotine and its structural analogue Phenylalanine amide individually and, in the blend, (50:50), as template materials in a Chitosan-Methacrylic acid functional monomer matrix. The temperature for polymerization is 60OC and time for polymerization, 12hrs (water bath heating), 4mins for (microwave heating). The characteristic thermal properties of the molecularly imprinted materials are investigated using Simultaneous Thermal Analysis (STA) profiling, while the absorption and separation efficiencies based on the relative retention times and peak areas of templates were studied amongst other properties. Transmission Electron Microscopy (TEM) results obtained, show the creation of heterogeneous nanocavities, regardless, the introduction of Caffeine a close structural analogue presented near-zero perfusion. This confirms the selectivity and specificity of the templated polymers despite its dual-templated nature. The STA results presented the materials as having decomposition temperatures above 250OC and a relative loss in mass of less than19% over a period within 50mins of heating. Consequent to this outcome, multi-templated systems can be fabricated to sequester specifically and selectively targeted toxicants in a mixed toxicant populated system effectively.

Keywords: chitosan, dual-templated, methacrylic acid, mixed-toxicants, molecularly-imprinted-polymer

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10917 Preparation and Characterization of TiO₂-SiO₂ Composite Films on Plastics Using Aqueous Peroxotitanium Acid Solution

Authors: Ayu Minamizawa, Jae-Ho Kim, Susumu Yonezawa

Abstract:

Aqueous peroxotitanium acid solution was prepared by the reaction between H₂O₂ solution and TiO₂ fluorinated using F₂ gas. The coating of TiO₂/SiO₂ multilayer on the surface of polycarbonate (PC) resin was carried out step by step using the TEOS solution and aqueous peroxotitanium acid solution. We confirmed each formation of SiO₂ and TiO₂ layer by scanning electron microscopy and energy-dispersive X-ray spectroscopy, and x-ray photoelectron spectroscopy results. The formation of a TiO₂ thin layer on SiO₂ coated on polycarbonate (PC) was carried out at 120 ℃ and for 15 min ~ 3 h with aqueous peroxotitanium acid solution using a hydrothermal synthesis autoclave reactor. The morphology TiO₂ coating layer largely depended on the reaction time, as shown in the results of SEM-EDS analysis. Increasing the reaction times, the TiO₂ layer expanded uniformly. Moreover, the surface fluorination of the SiO₂ layer can promote the formation of the TiO₂ layer on the surface.

Keywords: aqueous peroxotitanium acid solution, photocatalytic activity, polycarbonate, surface fluorination

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10916 Modification of Aliphatic-Aromatic Copolyesters with Polyether Block for Segmented Copolymers with Elastothemoplastic Properties

Authors: I. Irska, S. Paszkiewicz, D. Pawlikowska, E. Piesowicz, A. Linares, T. A. Ezquerra

Abstract:

Due to the number of advantages such as high tensile strength, sensitivity to hydrolytic degradation, and biocompatibility poly(lactic acid) (PLA) is one of the most common polyesters for biomedical and pharmaceutical applications. However, PLA is a rigid, brittle polymer with low heat distortion temperature and slow crystallization rate. In order to broaden the range of PLA applications, it is necessary to improve these properties. In recent years a number of new strategies have been evolved to obtain PLA-based materials with improved characteristics, including manipulation of crystallinity, plasticization, blending, and incorporation into block copolymers. Among the other methods, synthesis of aliphatic-aromatic copolyesters has been attracting considerable attention as they may combine the mechanical performance of aromatic polyesters with biodegradability known from aliphatic ones. Given the need for highly flexible biodegradable polymers, in this contribution, a series of aromatic-aliphatic based on poly(butylene terephthalate) and poly(lactic acid) (PBT-b-PLA) copolyesters exhibiting superior mechanical properties were copolymerized with an additional poly(tetramethylene oxide) (PTMO) soft block. The structure and properties of both series were characterized by means of attenuated total reflectance – Fourier transform infrared spectroscopy (ATR-FTIR), nuclear magnetic resonance spectroscopy (¹H NMR), differential scanning calorimetry (DSC), wide-angle X-ray scattering (WAXS) and dynamic mechanical, thermal analysis (DMTA). Moreover, the related changes in tensile properties have been evaluated and discussed. Lastly, the viscoelastic properties of synthesized poly(ester-ether) copolymers were investigated in detail by step cycle tensile tests. The block lengths decreased with the advance of treatment, and the block-random diblock terpolymers of (PBT-ran-PLA)-b-PTMO were obtained. DSC and DMTA analysis confirmed unambiguously that synthesized poly(ester-ether) copolymers are microphase-separated systems. The introduction of polyether co-units resulted in a decrease in crystallinity degree and melting temperature. X-ray diffraction patterns revealed that only PBT blocks are able to crystallize. The mechanical properties of (PBT-ran-PLA)-b-PTMO copolymers are a result of a unique arrangement of immiscible hard and soft blocks, providing both strength and elasticity.

Keywords: aliphatic-aromatic copolymers, multiblock copolymers, phase behavior, thermoplastic elastomers

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10915 Spectrophotometric Determination of Photohydroxylated Products of Humic Acid in the Presence of Salicylate Probe

Authors: Julide Hizal Yucesoy, Batuhan Yardimci, Aysem Arda, Resat Apak

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Humic substances produce reactive oxygene species such as hydroxyl, phenoxy and superoxide radicals by oxidizing in a wide pH and reduction potential range. Hydroxyl radicals, produced by reducing agents such as antioxidants and/or peroxides, attack on salicylate probe, and form 2,3-dihydroxybenzoate, 2,4-dihydroxybenzoate and 2,5-dihydroxybenzoate species. These species are quantitatively determined by using HPLC Method. Humic substances undergo photodegradation by UV radiation. As a result of their antioxidant properties, they produce hydroxyl radicals. In the presence of salicylate probe, these hydroxyl radicals react with salicylate molecules to form hydroxylated products (dihidroxybenzoate isomers). In this study, humic acid was photodegraded in a photoreactor at 254 nm (400W), formed hydroxyl radicals were caught by salicylate probe. The total concentration of hydroxylated salicylate species was measured by using spectrophotometric CUPRAC Method. And also, using results of time dependent experiments, kinetic of photohydroxylation was determined at different pHs. This method has been applied for the first time to measure the concentration of hydroxylated products. It allows to achieve the results easier than HPLC Method.

Keywords: CUPRAC method, humic acid, photohydroxylation, salicylate probe

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10914 Hedonic Pricing Model of Parboiled Rice

Authors: Roengchai Tansuchat, Wassanai Wattanutchariya, Aree Wiboonpongse

Abstract:

Parboiled rice is one of the most important food grains and classified in cereal and cereal product. In 2015, parboiled rice was traded more than 14.34 % of total rice trade. The major parboiled rice export countries are Thailand and India, while many countries in Africa and the Middle East such as Nigeria, South Africa, United Arab Emirates, and Saudi Arabia, are parboiled rice import countries. In the global rice market, parboiled rice pricing differs from white rice pricing because parboiled rice is semi-processing product, (soaking, steaming and drying) which affects to their color and texture. Therefore, parboiled rice export pricing does not depend only on the trade volume, length of grain, and percentage of broken rice or purity but also depend on their rice seed attributes such as color, whiteness, consistency of color and whiteness, and their texture. In addition, the parboiled rice price may depend on the country of origin, and other attributes, such as certification mark, label, packaging, and sales locations. The objectives of this paper are to study the attributes of parboiled rice sold in different countries and to evaluate the relationship between parboiled rice price in different countries and their attributes by using hedonic pricing model. These results are useful for product development, and marketing strategies development. The 141 samples of parboiled rice were collected from 5 major parboiled rice consumption countries, namely Nigeria, South Africa, Saudi Arabia, United Arab Emirates and Spain. The physicochemical properties and optical properties, namely size and shape of seed, colour (L*, a*, and b*), parboiled rice texture (hardness, adhesiveness, cohesiveness, springiness, gumminess, and chewiness), nutrition (moisture, protein, carbohydrate, fat, and ash), amylose, package, country of origin, label are considered as explanatory variables. The results from parboiled rice analysis revealed that most of samples are classified as long grain and slender. The highest average whiteness value is the parboiled rice sold in South Africa. The amylose value analysis shows that most of parboiled rice is non-glutinous rice, classified in intermediate amylose content range, and the maximum value was found in United Arab Emirates. The hedonic pricing model showed that size and shape are the key factors to determine parboiled rice price statistically significant. In parts of colour, brightness value (L*) and red-green value (a*) are statistically significant, but the yellow-blue value (b*) is insignificant. In addition, the texture attributes that significantly affect to the parboiled rice price are hardness, adhesiveness, cohesiveness, and gumminess. The findings could help both parboiled rice miller, exporter and retailers formulate better production and marketing strategies by focusing on these attributes.

Keywords: hedonic pricing model, optical properties, parboiled rice, physicochemical properties

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10913 A Thermodynamic Study of Parameters that Affect the Nitration of Glycerol with Nitric Acid

Authors: Erna Astuti, Supranto, Rochmadi, Agus Prasetya

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Biodiesel production from vegetable oil will produce glycerol as by-product about 10% of the biodiesel production. The amount of glycerol that was produced needed alternative way to handling immediately so as to not become the waste that polluted environment. One of the solutions was to process glycerol to polyglycidyl nitrate (PGN). PGN is synthesized from glycerol by three-step reactions i.e. nitration of glycerol, cyclization of 13- dinitroglycerine and polymerization of glycosyl nitrate. Optimum condition of nitration of glycerol with nitric acid has not been known. Thermodynamic feasibility should be done before run experiments in the laboratory. The aim of this study was to determine the parameters those affect nitration of glycerol and nitric acid and chose the operation condition. Many parameters were simulated to verify its possibility to experiment under conditions which would get the highest conversion of 1, 3-dinitroglycerine and which was the ideal condition to get it. The parameters that need to be studied to obtain the highest conversion of 1, 3-dinitroglycerine were mol ratio of nitric acid/glycerol, reaction temperature, mol ratio of glycerol/dichloromethane and pressure. The highest conversion was obtained in the range of mol ratio of nitric acid /glycerol between 2/1 – 5/1, reaction temperature of 5-25o C and pressure of 1 atm. The parameters that need to be studied further to obtain the highest conversion of 1.3 DNG are mol ratio of nitric acid/glycerol and reaction temperature.

Keywords: Nitration, glycerol, thermodynamic, optimum condition

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10912 Morphology and Permeability of Biomimetic Cellulose Triacetate-Impregnated Membranes: in situ Synchrotron Imaging and Experimental Studies

Authors: Amira Abdelrasoul

Abstract:

This study aimed to ascertain the controlled permeability of biomimetic cellulose triacetate (CTA) membranes by investigating the electrical oscillatory behavior across impregnated membranes (IM). The biomimetic CTA membranes were infused with a fatty acid to induce electrical oscillatory behavior and, hence, to ensure controlled permeability. In situ synchrotron radiation micro-computed tomography (SR-μCT) at the BioMedical Imaging and Therapy (BMIT) Beamline at the Canadian Light Source (CLS) was used to evaluate the main morphology of IMs compared to neat CTA membranes to ensure fatty acid impregnation inside the pores of the membrane matrices. A monochromatic beam at 20 keV was used for the visualization of the morphology of the membrane. The X-ray radiographs were recorded by means of a beam monitor AA-40 (500 μm LuAG scintillator, Hamamatsu, Japan) coupled with a high-resolution camera, providing a pixel size of 5.5 μm and a field of view (FOV) of 4.4 mm × 2.2 mm. Changes were evident in the phase transition temperatures of the impregnated CTA membrane at the melting temperature of the fatty acid. The pulsations of measured voltages were related to changes in the salt concentration of KCl in the vicinity of the electrode. Amplitudes and frequencies of voltage pulsations were dependent on the temperature and concentration of the KCl solution, which controlled the permeability of the biomimetic membranes. The presented smart biomimetic membrane successfully combined porous polymer support and impregnating liquid not only imitate the main barrier properties of the biological membranes but could be easily modified to achieve some new properties, such as facilitated and active transport, regulation by chemical, physical and pharmaceutical factors. These results open new frontiers for the facilitation and regulation of active transport and permeability through biomimetic smart membranes for a variety of biomedical and drug delivery applications.

Keywords: biomimetic, membrane, synchrotron, permeability, morphology

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10911 Performance Evaluation of Solid Lubricant Characteristics at Different Sliding Conditions

Authors: Suresh Kumar Reddy Narala, Rakesh Kumar Gunda

Abstract:

In modern industry, mechanical parts are subjected to friction and wear, leading to heat generation, which affects the reliability, life and power consumption of machinery. To overcome the tribological losses due to friction and wear, a significant portion of lubricant with high viscous properties allows very smooth relative motion between two sliding surfaces. Advancement in modern tribology has facilitated the use of applying solid lubricants in various industrial applications. Solid lubricant additives with high viscous thin film formation between the sliding surfaces can adequately wet and adhere to a work surface. In the present investigation, an attempt has been made to investigate and evaluate the tribological studies of various solid lubricants like MoS¬2, graphite, and boric acid at different sliding conditions. The base oil used in this study was SAE 40 oil with a viscosity of 220 cSt at 400C. The tribological properties were measured on pin-on-disc tribometer. An experimental set-up has been developed for effective supply of solid lubricants to the pin-disc interface zone. The results obtained from the experiments show that the friction coefficient increases with increase in applied load for all the considered environments. The tribological properties with MoS2 solid lubricant exhibit larger load carrying capacity than that of graphite and boric acid. The present research work also contributes to the understanding of the behavior of film thickness distribution of solid lubricant using potential contact technique under different sliding conditions. The results presented in this research work are expected to form a scientific basis for selecting the best solid lubricant in various industrial applications for possible minimization of friction and wear.

Keywords: friction, wear, temperature, solid lubricant

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10910 Effect of Antioxidants Addition in Combination with Milk Re Pasteurization on the Physical, Chemical and Sensory Properties of Condensed Yoghurt

Authors: Mahmoud Abu-Ghoush, Murad Al Holy

Abstract:

Our main goal in this project is to study the causes and finding solutions for both the hydrolytic and the oxidative rancidity that can be produced during condensed yoghurt production. The re pasteurization of the pasteurized milk and the addition of different types of antioxidants (ascorbic acid and propyl gallate) were used to achieve this goal. Chemical, physical, microbial and sensory tests were done to evaluate the product. It was found that there were significant differences between the different treatments and the control regarding the peroxide value. This means that the addition of both types of antioxidants have a positive effect in decreasing the rancidity value. However, it was found that there were some samples have hydrolytic rancidity flavour without any type of oxidative rancidity (low peroxide value). To overcome this problem the re pasteurization step was used to destroy all the vegetative form of microbes. It was found that this treatment was very useful in controlling the rancidity flavour according to the sensory evaluation of the condensed yoghurt products for several batches. The best condensed yoghurt which contains 0.25% ascorbic acid exhibited the highest sensory properties values. Also, it has the ability in lowering the oxidative rancidity in the combination with the re pasteurization step of the pasteurized milk. This suggests that a higher quality and stable condensed yoghurt can be obtained upon using this combination. These results may help producers in selecting the best treatment methods to overcome the rancidity flavor in this type of condensed yoghurt.

Keywords: antioxidants, condensed yoghurt, repasturization, condensed milk

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10909 Introduction of PMMA-Tag to VHH for Improving Recovery and Immobilization Rate of VHHS

Authors: Bongmun Kang, Kagnari Yamakawa, Yoshihisa Hagihara, Yuji Ito, Michimasa Kishimoto, Yoichi Kumada

Abstract:

The PMMA-tag was genetically fused with the C-terminal region of VHH molecules. This antibody, VHH, is known as a single-chain domain, which is devoid of light chains. The PMMA-tag, which could affect the isoelectric point (pI) changeable with a charge of amino acid in VHHs were closely related to the solubility of VHH molecules during refolding. The genetic fusion of PMMA-tag to C-terminal region of VHHs significantly affects the recovery of their soluble protein during refolding by 50 mM TAPS at pH 8.5. It could be refolded with a recovery of more than 95% by dialysis at pH 8.5. A marked difference in the antigen-binding activities in the adsorption state was significantly high in VHH-PM compared to the wild type of VHH. There are approximately 8-fold differences in the antigen-binding activities in the adsorption state between VHH-PM and VHH.

Keywords: VHH, PMMA-tag, isoelectric point, pH, Solubility, refolding, immobilization, ELISA

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10908 Physicochemical Properties and Toxicity Studies on a Lectin from the Bulb of Dioscorea bulbifera

Authors: Uchenna Nkiruka Umeononihu, Adenike Kuku, Oludele Odekanyin, Olubunmi Babalola, Femi Agboola, Rapheal Okonji

Abstract:

In this study, a lectin from the bulb of Dioscorea bulbifera was purified, characterised, and its acute and sub-acute toxicity was investigated with a view to evaluate its toxic effects in mice. The protein from the bulb was extracted by homogenising 50 g of the bulb in 500 ml of phosphate buffered saline (0.025 M) of pH 7.2, stirred for 3 hr, and centrifuged at the speed of 3000 rpm. Blood group and sugar specificity assays of the crude extract were determined. The lectin was purified in a two-step procedure- gel filtration on Sephadex G-75 and affinity chromatography on Sepharose 4-B arabinose. The degree of purity of the purified lectin was ascertained by SDS-polyacrylamide gel electrophoresis. Detection of covalently bound carbohydrate was carried out with Periodic Acid-Schiffs (PAS) reagent staining technique. Effects of temperature, pH, and EDTA on the lectin were carried out using standard methods. This was followed by acute toxicity studies via oral and subcutaneous routes using mice. The animals were monitored for mortality and signs of toxicity. The sub-acute toxicity studies were carried out using rats. Different concentrations of the lectin were administered twice daily for 5 days via the subcutaneous route. The animals were sacrificed on the sixth day; blood samples and liver tissues were collected. Biochemical assays (determination of total protein, direct bilirubin, Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), catalase (CAT), and superoxide dismutase (SOD)) were carried out on the serum and liver homogenates. The collected organs (heart, liver, kidney, and spleen) were subjected to histopathological analysis. The results showed that lectin from the bulbs of Dioscorea bulbifera agglutinated non-specifically the erythrocytes of the human ABO system as well as rabbit erythrocytes. The haemagglutinating activity was strongly inhibited by arabinose and dulcitol with minimum inhibitory concentrations of 0.781 and 6.25, respectively. The lectin was purified to homogeneity with native and subunit molecular weights of 56,273 and 29,373 Daltons, respectively. The lectin was thermostable up to 30 0C and lost 25 %, 33.3 %, and 100 % of its heamagglutinating activity at 40°C, 50°C, and 60°C, respectively. The lectin was maximally active at pH 4 and 5 but lost its total activity at pH eight, while EDTA (10 mM) had no effect on its haemagglutinating activity. PAS reagent staining showed that the lectin was not a glycoprotein. The sub-acute studies on rats showed elevated levels of ALT, AST, serum bilirubin, total protein in serum and liver homogenates suggesting damage to liver and spleen. The study concluded that the aerial bulb of D. bulbifera lectin was non-specific in its heamagglutinating activity and dimeric in its structure. The lectin shared some physicochemical characteristics with lectins from other Dioscorecea species and was moderately toxic to the liver and spleen of treated animals.

Keywords: Dioscorea bulbifera, heamagglutinin, lectin, toxicity

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