Search results for: enzymatic biosensor

Authors: Waranya Imprasittichai, Patsarawadee Paojinda, Sudaratana R. Krungkrai, Nirianne Marie Q. Palacpac, Toshihiro Horii, Jerapan Krungkrai

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Fusion of the last two enzymes in the pyrimidine biosynthetic pathway in the inversed order by having COOH-terminal orotate phosphoribosyltransferase (OPRT) and NH2-terminal orotidine 5'-monophosphate decarboxylase (OMPDC), as OMPDC-OPRT, are described in many organisms. In this study, we constructed gene fusions of Plasmodium falciparum OMPDC-OPRT (1,836 bp) in pTrcHisA vector and expressed as an 6xHis-tag bifunctional protein in three Escherichia coli strains (BL21, Rosetta, TOP10) at 18 °C, 25 °C and 37 °C. The recombinant bifunctional protein was partially purified by Ni-Nitrilotriacetic acid-affinity chromatography. Specific activities of OPRT and OMPDC domains in the bifunctional enzyme expressed in E. coli TOP10 cells were approximately 3-4-fold higher than those in BL21 cells. There were no enzymatic activities when the construct vector expressed in Rosetta cells. Maximal expression of the fused gene was observed at 18 °C and the bifunctional enzyme had specific activities of OPRT and OMPDC domains in a ratio of 1:2. These results provide greater yields and better catalytic activities of the bifunctional OMPDC-OPRT enzyme for further purification and kinetic study.

Keywords: bifunctional enzyme, orotate phosphoribosyltransferase, orotidine 5'-monophosphate decarboxylase, plasmodium falciparum

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Authors: Elena G. Salina, Laurent R. Chiarelli, Maria R. Pasca, Vadim A. Makarov

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Tuberculosis is one of the most challenging threats to human health, confronted by the problem of drug resistance. Evidently, new drugs for tuberculosis are urgently needed. Thienopyrimidine TP053 is one of the most promising new antitubercular prodrugs. Mycothiol-dependent reductase Mrx2, encoded by rv2466c, is known to be a TP053 activator; however, the precise mode of action of this compound remained unclear. Being highly active against both replicating and non-replicating tuberculosis bacilli, TP053 also revealed dose-escalating activity for M. tuberculosis-infected murine macrophages. The chemical structure of TP053 is characterized by the presence of NO₂ group which was suggested to be responsible for the toxic effects of the activated compound. Reduction of a nitroaromatic moiety of TP53 by Mrx2 was hypothesized to result in NO release. Analysis of the products of enzymatic activation of TP053 by Mrx2 by the Greiss reagent clearly demonstrated production of nitric oxide in a time-dependent manner. Mass-spectra of cell lysates of TP-treated M. tuberculosis bacilli demonstrated the transformation of TP053 to its non-active metabolite with Mw=261 that corresponds NO release. The mechanism of NO toxicity for bacteria includes DNA damage and degradation of iron-sulfur centers, especially under oxygen depletion. Thus, TP-053 drug-like scaffold is prospective for further development of novel anti-TB drug. This work was financially supported by the Russian Foundation for Basic Research (Grant 17-04-00342).

Keywords: drug discovery, M. tuberculosis, nitric oxide, NO donors

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Authors: Ka Ho Yau, Jan Frederick Engels, Kwok Kei Lai, Reinhard Renneberg

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Lateral flow test is a prevalent technology in various sectors such as food, pharmacology and biomedical sciences. Colloidal gold (CG) is widely used as the signalling molecule because of the ease of synthesis, bimolecular conjugation and its red colour due to intrinsic SPRE. However, the production of colloidal gold is costly and requires vigorous conditions. The stability of colloidal gold are easily affected by environmental factors such as pH, high salt content etc. Silica nanoparticles are well known for its ease of production and stability over a wide range of solvents. Using reverse micro-emulsion (w/o), silica nanoparticles with different sizes can be produced precisely by controlling the amount of water. By incorporating different water-soluble dyes, a rainbow colour of the silica nanoparticles could be produced. Conjugation with biomolecules such as antibodies can be achieved after surface modification of the silica nanoparticles with organosilane. The optimum amount of the antibodies to be labelled was determined by Bradford Assay. In this work, we have demonstrated the ability of the dye-doped silica nanoparticles as a signalling molecule in lateral flow test, which showed a semi-quantitative measurement of the analyte. The image was further analysed for the LOD=10 ng of the analyte. The working range and the linear range of the test were from 0 to 2.15μg/mL and from 0 to 1.07 μg/mL (R2=0.988) respectively. The performance of the tests was comparable to those using colloidal gold with the advantages of lower cost, enhanced stability and having a wide spectrum of colours. The positives lines can be imaged by naked eye or by using a mobile phone camera for a better quantification. Further research has been carried out in multicolour detection of different biomarkers simultaneously. The preliminary results were promising as there was little cross-reactivity being observed for an optimized system. This approach provides a platform for multicolour detection for a set of biomarkers that enhances the accuracy of diseases diagnostics.

Keywords: colorimetric detection, immunosensor, paper-based biosensor, silica

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Authors: Huda Aldossari

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Members of the Brassicaceae family, such as rocket species, have high concentrations of glucosinolates (GLSs). GSLs and isothiocyanates (ITCs), the product of GLSs hydrolysis, are the most influential compounds that affect flavour in rocket species. Aside from their contribution to the flavour, GSLs and ITCs are of particular interest due to their potential ability to inhibit the growth of human pathogenic bacteria such as E. coli O157. Quantitative and qualitative analysis of glucosinolate compounds in rocket extracts was obtained by Liquid Chromatography-Mass Spectrometry (LC–MS).Each individual component of non-volatile GLSs and ITCs was isolated by High-Performance Liquid Chromatography (HPLC) fractionation. The identity and purity of each fraction were confirmed using Ultra High-Performance Liquid Chromatography (UPLC). The separation of glucosinolates in the complex rocket extractions was performed by optimizing a HPLC fractionation method through changing the mobile phase composition, solvent gradient, and the flow rate. As a result, six glucosinolates compounds (Glucosativin, 4-Methoxyglucobrassicin, Glucotropaeolin GTP, Glucoiberin GIB, Diglucothiobenin, and Sinigrin) have been isolated, identified and quantified in the complex samples. This step aims to evaluate the antibacterial activity of glucosinolates and their enzymatic hydrolysis against bacterial growth of E.coli k12. Therefore, fractions from this study will be used to determine the most active compounds by investigating the efficacy of each component of GLSs and ITCs at inhibiting bacterial growth.

Keywords: rocket, glucosinolates, E.coli k12., HPLC fractionatio

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Authors: Senem Suna, Canan Ece Tamer, Ömer Utku Çopur

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In this research, dried linden (Tilia argentea) leaves and blossoms were used as a raw material for mineral enriched herbal tea beverage production. For this aim, %1 dried linden was infused with boiling water (100 °C) for 5 minutes. After cooling, sucrose, citric acid, ascorbic acid, natural lemon flavor and natural mineral water were added. Beverage samples were plate filtered, filled into 200-mL glass bottles, capped then pasteurized at 98 °C for 15 minutes. Water soluble dry matter, titratable acidity, ascorbic acid, pH, minerals (Fe, Ca, Mg, K, Na), color (L*, a*, b*), turbidity, bioaccessible phenolics and antioxidant capacity were analyzed. Water soluble dry matter, titratable acidity, and ascorbic were determined as 7.66±0.28 g/100 g, 0.13±0.00 g/100 mL, and 19.42±0.62 mg/100 mL, respectively. pH was measured as 3.69. Fe, Ca, Mg, K and Na contents of the beverage were determined as 0.12±0.00, 115.48±0.05, 34.72±0.14, 48.67±0.43 and 85.72±1.01 mg/L, respectively. Color was measured as 13.63±0.05, -4.33±0.05, and 3.06±0.05 for L*, a*, and b* values. Turbidity was determined as 0.69±0.07 NTU. Bioaccessible phenolics were determined as 312.82±5.91 mg GAE/100 mL. Antioxidant capacities of chemical (MetOH:H₂O:HCl) and physiological extracts (in vitro digestive enzymatic extraction) with DPPH (27.59±0.53 and 0.17±0.02 μmol trolox/mL), FRAP (21.01±0.97 and 13.27±0.19 μmol trolox/mL) and CUPRAC (44.71±9.42 and 2.80±0.64 μmol trolox/mL) methods were also evaluated. As a result, enrichment with natural mineral water was proposed for the development of functional and nutritional values together with a good potential for commercialization.

Keywords: linden, herbal tea beverage, bioaccessibility, antioxidant capacity

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Authors: J. B. Minari, O. B. Oloyede

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Polymorphonuclear neutrophils (PMNs) play a crucial role in a variety of infections caused by bacteria, fungi, and parasites. Indeed, the involvement of PMNs in host defence against Plasmodium falciparum is well documented both in vitro and in vivo. Many of the antimalarial drugs such as chloroquine used in the treatment of human malaria significantly reduce the immune response of the host in vitro and in vivo. Myeloperoxidase is the most abundant enzyme found in the polymorphonuclear neutrophil which plays a crucial role in its function. This study was carried out to investigate the effect of chloroquine on the enzyme. In investigating the effects of the drug on myeloperoxidase, the influence of concentration, pH, partition ratio estimation and kinetics of inhibition were studied. This study showed that chloroquine is concentration-dependent inhibitor of myeloperoxidase with an IC50 of 0.03 mM. Partition ratio estimation showed that 40 enzymatic turnover cycles are required for complete inhibition of myeloperoxidase in the presence of chloroquine. The influence of pH on the effect of chloroquine on the enzyme showed significant inhibition of myeloperoxidase at physiological pH. The kinetic inhibition studies showed that chloroquine caused a non-competitive inhibition with an inhibition constant Ki of 0.27mM. The results obtained from this study shows that chloroquine is a potent inhibitor of myeloperoxidase and it is capable of inactivating the enzyme. It is therefore considered that the inhibition of myeloperoxidase in the presence of chloroquine as revealed in this study may partly explain the impairment of polymorphonuclear neutrophil and consequent immunosuppression of the host defence system against secondary infections.

Keywords: myeloperoxidase, chloroquine, inhibition, neutrophil, immune

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

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

Keywords: paraquat, paraquat nanoparticles, liver, oxidative stress

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Authors: Shringika Soni, Utkarsh Jain, Nidhi Chauhan

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Aptamers are recently discovered ~80-100 bp long artificial oligonucleotides that not only demonstrated their applications in therapeutics; it is tremendously used in diagnostic and sensing application to detect different biomarkers and drugs. Synthesizing aptamers for proteins or genomic template is comparatively feasible in laboratory, but drugs or other chemical target based aptamers require major specification and proper optimization and validation. One has to optimize all selection, amplification, and characterization steps of the end product, which is extremely time-consuming. Therefore, we performed asymmetric PCR (polymerase chain reaction) for random oligonucleotides pool synthesis, and further use them in Systematic evolution of ligands by exponential enrichment (SELEX) for anti-psychotic drugs based aptamers synthesis. Anti-psychotic drugs are major tranquilizers to control psychosis for proper cognitive functions. Though their low medical use, their misuse may lead to severe medical condition as addiction and can promote crime in social and economical impact. In this work, we have approached the in-vitro SELEX method for ssDNA synthesis for anti-psychotic drugs (in this case ‘target’) based aptamer synthesis. The study was performed in three stages, where first stage included synthesis of random oligonucleotides pool via asymmetric PCR where end product was analyzed with electrophoresis and purified for further stages. The purified oligonucleotide pool was incubated in SELEX buffer, and further partition was performed in the next stage to obtain target specific aptamers. The isolated oligonucleotides are characterized and quantified after each round of partition, and significant results were obtained. After the repetitive partition and amplification steps of target-specific oligonucleotides, final stage included sequencing of end product. We can confirm the specific sequence for anti-psychoactive drugs, which will be further used in diagnostic application in clinical and forensic set-up.

Keywords: anti-psychotic drugs, aptamer, biosensor, ssDNA, SELEX

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Authors: Seyyed Mohammad Amin Mousavi Sagharchi, Alireza Mahmoudi Nasab, Tim Bakker

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Introduction: Mycobacterium tuberculosis (MTB) is the most intelligent bacterium that existed in the world to our best knowledge. This bacterium can cause tuberculosis (TB) which is responsible for its spread speed and murder of millions of people around the world. MTB has the practical function to escape from anti-tuberculosis drugs (AT), for this purpose, it handles some mutations in the main genes and creates new patterns for inhibited genes. Method and materials: Researchers have their best tries to safely isolate MTB from the sputum specimens of 35 patients in some hospitals in the Tehran province and detect MTB by culture on Löwenstein-Jensen (LJ) medium and microscopic examination. DNA was extracted from the established bacterial colony by enzymatic extraction method. It was amplified by the polymerase chain reaction (PCR) method, reverse hybridization, and evaluation for detection of resistance genes; generally, researchers apply GenoType MTBDRplus assay. Results: Investigations of results declare us that 21 of the isolated specimens (about 60%) have mutation in rpoB gene, which resisted to rifampicin (most prevalence), and 8 of them (about 22.8%) have mutation in katG or inhA genes which resisted to isoniazid. Also, 4 of them (about 11.4%) don't have any mutation, and 2 of them (about 5.7%) have mutation in every three genes, which makes them resistant to the two drugs mentioned above. Conclusion: Rifampicin and isoniazid are two essential AT that using in the first line of treatment. Resistance in rpoB, and katG, and inhA genes related to mentioned drugs lead to ineffective treatment.

Keywords: mycobacterium tuberculosis, tuberculosis, drug resistance, isoniazid, rifampicin

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Authors: Su Yin Chiam, Qing Xin Zhang, Jaehoon Chung

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Complementary metal-oxide-semiconductor (CMOS) integrated circuits (ICs) have obtained increased attention in the biosensor community because CMOS technology provides cost-effective and high-performance signal processing at a mass-production level. In order to supply biological samples and reagents effectively to the sensing elements, there are increasing demands for seamless integration of microfluidic components on the fabricated CMOS wafers by post-processing. Although the PDMS microfluidic channels replicated from separately prepared silicon mold can be typically aligned and bonded onto the CMOS wafers, it remains challenging owing the inherently limited aligning accuracy ( > ± 10 μm) between the two layers. Here we present a new post-processing method to create three-dimensional microfluidic components using two different polarities of photoresists, an epoxy-based negative SU-8 photoresist and positive SPR220-7 photoresist. The positive photoresist serves as a sacrificial layer and the negative photoresist was utilized as a structural material to generate three-dimensional structures. Because both photoresists are patterned using a standard photolithography technology, the dimensions of the structures can be effectively controlled as well as the alignment accuracy, moreover, is dramatically improved (< ± 2 μm) and appropriately can be adopted as an alternative post-processing method. To validate the proposed processing method, we applied this technique to build cell-trapping structures. The SU8 photoresist was mainly used to generate structures and the SPR photoresist was used as a sacrificial layer to generate sub-channel in the SU8, allowing fluid to pass through. The sub-channel generated by etching the sacrificial layer works as a cell-capturing site. The well-controlled dimensions enabled single-cell capturing on each site and high-accuracy alignment made cells trapped exactly on the sensing units of CMOS biosensors.

Keywords: SU-8, microfluidic, MEMS, microfabrication

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Authors: Ana Tsokilauri

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The uppermost factor in the regulation of lignin-cellulose activity of decaying white rot or free rot are the substances serving as carbon and nitrogen nutrition of microorganisms and are considered as the most important factor of generative activity of white rot. The research object was Basidiomycete Fungi, peculiar and common in Georgia, and the separation of 10 of them as pure crops. The unidentified pure crops have tasted in order to be determined the potential of synthesis of lignin-degrading enzymes and the substrate of optimal lignocellulose growth. One of the most important aspects of the research conducted on Basidiomycetes was the use of specific lignocellulosic residues for selecting Fungi as a substrate of their growth. In order to increase lignocellulose with the help of substrate, crops were selected from the screening stage that showed good laccase activity. (Dusheti 1; Dusheti 10; Fshavi 5; Fshavi1; Fshavi 8; Fshavi 32; Manglisi 26; Sabaduri20; Dusheti 7; Sabaduri 1 ), Among the selected cultures, the crops with good laccase activity against the following substances, in particular: Dusheti 1- in this case, the rate of enzymatic activity on bran substrate was -105,6 u/ml, mandarin-96,4 u/ml. In case of corn - 102,9 u/ml. In case of Dusheti 7- the indicators were as follows: bananas-121,7 u/ml, mandarin-125,4 u/ml, corn - 117,1 u/ml. In the case of Sanaduri 32, the laccase activity was as follows: pomegranate- 101,2 u/ml. As a result of conducted experiments, the synthesis and activity rates of enzymes depending on plant substrates varied within a fairly wide range, which is still being under research.

Keywords: Lignocellulosic substrate, Basidiomycetes, white-rot basidiomycetes, Laccase

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Authors: Lela Pintarić, Iva Rezić, Ana Vrsalović Presečki

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Considering an enormous increase of the use of metal nanoparticles with the exactly defined characteristics, the main goal of this research was to found the optimal and environmental friendly method of their synthesis. The synthesis of the inorganic core-shell nanoparticles was optimized as a model. The core-shell nanoparticles are composed of the enzyme core belted with the metal ions, oxides or salts as a shell. In this research, enzyme urease was the core catalyst and the shell nanoparticle was made of silver. Silver nanoparticles are widespread utilized and some of their common uses are: as an addition to disinfectants to ensure an aseptic environment for the patients, as a surface coating for neurosurgical shunts and venous catheters, as an addition to implants, in production of socks for diabetics and athletic clothing where they improve antibacterial characteristics, etc. Characteristics of synthesized nanoparticles directly depend on of their size, so the special care during this optimization was given to the determination of the size of the synthesized nanoparticles. For the purpose of the above mentioned optimization, sixteen experiments were generated by the Design of Experiments (DoE) method and conducted under various temperatures, with different initial concentration of the silver nitrate and constant concentration of the urease of two separate manufacturers. Synthesized nanoparticles were analyzed by the Nanoparticle Tracking Analysis (NTA) method on Malvern NanoSight NS300. Results showed that the initial concentration of the silver ions does not affect the concentration of the synthesized silver nanoparticles neither their size distribution. On the other hand, temperature of the experiments has affected both of the mentioned values.

Keywords: core-shell nanoparticles, optimization, silver, urease

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Authors: Arredondo Valdés Roberto, Hernández Castillo Francisco Daniel, Laredo Alcalá Elan Iñaky, Gonzalez Gallegos Esmeralda, Castro Del Angel Epifanio

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The common bean or Phaseolus vulgaris L. is the most important food legume for direct consumption in the world. Fusarium dry rot in the major fungus disease affects Phaseolus vulgaris L, after planting. In another hand, Rhizoctonia can be found on all underground parts of the plant and various times during the growing season. In recent years, the world has conducted studies about the use of natural products as substitutes for herbicides and pesticides, because of possible ecological and economic benefits. Plants respond to fungal invasion by activating defense responses associated with accumulation of several enzymes and inhibitors, which prevent pathogen infection. This study focused on the role of proteins, peroxidase (POD), phenylalanine ammonia lyase (PAL), in imparting resistance to soft rot pathogens by applied different bacterial consortium, formulated and provided by Biofertilizantes de Méxicanos industries, analyzing the enzyme activity at different times of application (6 h, 12 h and 24 h). The resistance of these treatments was correlated with high POD and PAL enzyme activity as well as increased concentrations of proteins. These findings show that PAL, POD and synthesis of proteins play a role in imparting resistance to Phaseolus vulgaris L. soft rot infection by Fusarium and Rhizoctonia.

Keywords: fusarium, peroxidase, phenylalanine ammonia lyase, rhizoctonia

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Authors: D. M. El-Tanbouly, R. M. Abdelsalam, A. S. Attia, M. T. Abdel-Aziz

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Lipopolysaccharide (LPS) endotoxin, a component of the outer membrane of Gram-negative bacteria, is involved in the pathogenesis of sepsis. LPS administration induces systemic inflammation that mimics many of the initial clinical features of sepsis and has deleterious effects on several organs including the liver and eventually leading to septic shock and death. The present study aimed to investigate the protective effect of magnesium, a well-known cofactor in many enzymatic reactions and a critical component of the antioxidant system, on hepatic damage associated with LPS induced- endotoxima in mice. Mg (20 and 40 mg/kg, po) was administered for 7 consecutive days. Systemic inflammation was induced one hour after the last dose of Mg by a single dose of LPS (2 mg/kg, ip) and three hours thereafter plasma was separated, animals were sacrificed and their livers were isolated. LPS-treated mice suffered from hepatic dysfunction revealed by histological observation, elevation in plasma transaminases activities, C-reactive protein content and caspase-3, a critical marker of apoptosis. Liver inflammation was evident by elevation in liver cytokines contents (TNF-α and IL-10) and myeloperoxidase (MPO) activity. Additionally, oxidative stress was manifested by increased liver lipoperoxidation, glutathione depletion, elevated total nitrate/nitrite (NOx) content and glutathione peroxidase (GPx) activity. Pretreatment with Mg largely mitigated these alternations through its anti-inflammatory and antioxidant potentials. Mg, therefore, could be regarded as an effective strategy for prevention of liver damage associated with septicemia.

Keywords: LPS, liver damage, magnesium, septicemia

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Authors: Isaac Gbadura Adanlawo, Olusola Olalekan Elekofehinti

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This work investigated the antihyperglycemic, antioxidant and antihyperlipidemic effects of saponins from the fruit of Solanum anguivi, a plant generally used in folk medicine to treat diabetes and hypertension and to compare its effect with metformin in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in albino rats by administration of STZ (65 mg/kg) intraperitoneally. Saponin (40 and 100 mg/kg) was administered by oral gavage once daily for 21 days. Metformin (200 mg/kg b.w.) was administered as the positive control. The effect of saponin on blood glucose, serum lipids and enzymatic antioxidants defense systems, like superoxide dismutase (SOD), catalase (CAT), as well as MDA levels in serum, liver and pancreas were studied. Saponins from S. anguivi fruits reduced the blood glucose, total cholesterol (TC), triglycerides (TG) and low-density lipoprotein (LDL) levels in STZ-diabetic rats. They also significantly abolished the increase in MDA level in serum, liver and pancreas of diabetic rats. The activities of SOD and CAT in serum, liver and pancreas were significantly increased as well as concentration of HDL in the serum. Metformin had the same effect as saponin but saponins seems to be more potent in reducing serum TC, TG, LDL, and MDA, and increasing SOD and CAT. Conclusions: These results suggest that saponins from S. anguivi fruits have anti-diabetic and antihypercholesterolemic, antihypertriglyceridemic antiperoxidative activities mediated through their antioxidant properties. Also, saponins appeared to have more hypolipidemic, antiperoxidative and antioxidant activity than metformin.

Keywords: saponin, diabetes, metformin, streptozotocin, Solanum anguivi

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Authors: Nivedita Sharma

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The global demand for fossil fuels is very high, but their use is not sustainable since its reserves are declining. Additionally, fossil fuels are responsible for the accumulation of greenhouse gases. The emission of greenhouse gases from the transport sector can be reduced by substituting fossil fuels by biofuels. Thus, renewable fuels capable of sequestering carbon dioxide are in high demand. Second‐generation biofuels, which require lignocellulosic biomass as a substrate and ultimately producing ethanol, fall largely in this category. Bioethanol is a favorable and near carbon-neutral renewable biofuel leading to reduction in tailpipe pollutant emission and improving the ambient air quality. Lignocellulose consists of three main components: cellulose, hemicellulose and lignin which can be converted to ethanol with the help of microbial enzymes. Enzymatic hydrolysis of lignocellulosic biomass in 1st step is considered as the most efficient and least polluting methods for generating fermentable hexose and pentose sugars which subsequently are fermented to power alcohol by yeasts in 2nd step of the process. In the present technology, a complete bioconversion process i.e. potential hydrolytic enzymes i.e. cellulase and xylanase producing microorganisms have been isolated from different niches, screened for enzyme production, identified using phenotyping and genotyping, enzyme production, purification and application of enzymes for saccharification of different lignocellulosic biomass followed by fermentation of hydrolysate to ethanol with high yield is to be presented in detail.

Keywords: cellulase, xylanase, lignocellulose, bioethanol, microbial enzymes

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Authors: Renata Shitakubo, Hanyu Yangcheng, Jay-lin Jane, Fernanda Peroni Okita, Beatriz Cordenunsi

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In the degradation transitory starch model, the enzymatic activity of glucan/water dikinase (GWD) and phosphoglucan/water dikinase (PWD) are essential for the granule degradation. GWD and PWD phosphorylate glucose molecules in the positions C6 and C3, respectively, in the amylopectin chains. This action is essential to allow that β-amylase degrade starch granules without previous action of α-amylase. During banana starch degradation, as part of banana ripening, both α- and β-amylases activities and proteins were already detected and, it is also known that there is a GWD and PWD protein bounded to the starch granule. Therefore, the aim of this study was to quantify both Gluc-6P and Gluc-3P in order to estimate the importance of the GWD-PWD-β-amylase pathway in banana starch degradation. Starch granules were isolated as described by Peroni-Okita et al (Carbohydrate Polymers, 81:291-299, 2010), from banana fruit at different stages of ripening, green (20.7%), intermediate (18.2%) and ripe (6.2%). Total phosphorus content was determinate following the Smith and Caruso method (1964). Gluc-6P and Gluc-3P quantifications were performed as described by Lim et al (Cereal Chemistry, 71(5):488-493, 1994). Total phosphorous content in green banana starch is found as 0.009%, intermediary banana starch 0.006% and ripe banana starch 0.004%, both by the colorimetric method and 31P-NMR. The NMR analysis showed the phosphorus content in C6 and C3. The results by NMR indicate that the amylopectin is phosphorylate by GWD and PWD before the bananas become ripen. Since both the total content of phosphorus and phosphorylated glucose molecules at positions C3 and C6 decrease with the starch degradation, it can be concluded that this phosphorylation occurs only in the surface of the starch granule and before the fruit be harvested.

Keywords: starch, GWD, PWD, 31P-NMR

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Authors: Emmanuel S. Adeyanju

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This study assessed the effects of continuous training on anthropometric characteristics of adolescents in Kano, Nigeria. The anthropometric measures of per cent body fat (%BF), body mass index (BMI), conicity index (CI) and waist-to-hip ratio (WHR) were selected because of their roles in increased adiposity and favourable cardiovascular disease (CVD) factor profiles in children and adolescence. The international standards and procedures were followed in all the measurements. A total of thirty (30) subjects (M=15; F=15), selected at random, were divided into two groups; one training (M=10; F=10) and the other control (M=5; F=5). Both groups were tested before training, at six (6) and 12 weeks in all the listed variables. The training group had 12 weeks continuous training which involved running round the standard 400 m track of the college following standard procedures; while the control group did not. The findings revealed significant sex-specific reductions in %BF (F=610.482 ˂ 0.05), BMI (F=73.860 ˂ 0.05), WHR (F=49.756 ˂ 0.05); however, no significant training effect on CI (F=1.855 ˃ 0.05) and WHR (F=1.956 ˃ 0.05) was found. Greater modifications found in females than in males (except in CI and WHR) due to training were probably related to their initial level of fitness and enzymatic modifications at subcellular level during training. The result also revealed significant relationship between the modifications in %BF, BMI and WHR but failed to establish any between CI and other adiposity measures. Thus, to avert the consequences of obesity and overweight, the declining fitness level of adolescents should be checked by ensuring they engaged in regular moderate-to-vigorous physical activity (MVPA) programmes. Such a childhood habit of exercise developed early in life will have a carry-over value into adult life and improve the quality of adult population.

Keywords: adiposity, anthropometry, conicity, continuous training

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Authors: Nesrine Lenchi, Salima Kebbouche-Gana, Laddada Belaid, Mohamed Lamine Khelfaoui, Mohamed Lamine Gana

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Saline lakes are extreme hypersaline environments that are considered five to ten times saltier than seawater (150 – 300 g L-1 salt concentration). Hypersaline regions differ from each other in terms of salt concentration, chemical composition and geographical location, which determine the nature of inhabitant microorganisms. In order to explore the diversity of moderate and extreme halophiles Bacteria in Chott Tinsilt (East of Algeria), an isolation program was performed. In the first time, water samples were collected from the saltern during pre-salt harvesting phase. Salinity, pH and temperature of the sampling site were determined in situ. Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions. Isolates were obtained by plating out the samples in complex and synthetic media. In this study, seven halophiles cultures of Bacteria were isolated. Isolates were studied for Gram’s reaction, cell morphology and pigmentation. Enzymatic assays (oxidase, catalase, nitrate reductase and urease), and optimization of growth conditions were done. The results indicated that the salinity optima varied from 50 to 250 g L-1, whereas the optimum of temperature range from 25°C to 35°C. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. The results showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Idiomarina, Halobacillus Thalassobacillus and Planococcus genera and what may represent a new bacterial genus.

Keywords: bacteria, Chott, halophilic, 16S rRNA

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Authors: Vinod Nair, C. Sadasivan

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Beta-lactam antibiotics are the most frequently prescribed medications in modern medicine. The antibiotic resistance by the production of enzyme beta-lactamase is an important mechanism seen in microorganisms. Resistance to beta-lactams mediated by beta-lactamases can be overcome successfully with the use of beta-lactamase inhibitors. New generations of the antibiotics contain mostly synthetic compounds, and many side effects have been reported for them. Combinations of beta-lactam and beta-lactamase inhibitors have become one of the most successful antimicrobial strategies in the current scenario of bacterial infections. Plant-based drugs are very cheap and having lesser adverse effect than synthetic compounds. The synergistic effect of eugenol acetate with beta-lactams restores the activity of beta-lactams, allowing their continued clinical use. It is reported here the enhanced inhibitory effect of phytochemical, eugenol acetate, isolated from the plant Syzygium aromaticum with beta-lactams on beta-lactamase. The compound was found to have synergistic effect with the antibiotic amoxicillin against antibiotic-resistant strain of S.aureus. The enzyme was purified from the organism and incubated with the compound. The assay showed that the compound could inhibit the enzymatic activity of beta-lactamase. Modeling and molecular docking studies indicated that the compound can fit into the active site of beta-lactamase and can mask the important residue for hydrolysis of beta-lactams. The synergistic effects of eugenol acetate with beta-lactam antibiotics may justify, the use of these plant compounds for the preparation of β-lactamase inhibitors against β-lactam resistant S.aureus.

Keywords: betalactamase, eugenol acetate, synergistic effect, molecular modeling

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Authors: Jasna M. Čanadanović-Brunet, Gordana S. Ćetković, Jelena J. Vulić, Sonja M. Djilas, Vesna T. Tumbas Šaponjac, Sladjana M. Stajčić

Abstract:

Honey serves as a source of natural antioxidants, which are effective in reducing the risk of heart disease, cancer, immune-system decline, cataracts, different inflammatory processes, and also prevent deteriorative oxidation reactions in foods such as enzymatic browning of fruit and vegetables. Honey is a natural saturated sugar solution, but it also contains certain minor constituents, proteins, enzymes, amino and organic acids, lipids, vitamins, phenolic acids, flavonoids and carotenoids. It is consumed in its natural form alone, but also in combination with nuts and various kinds of dried fruits. The aim of this research was to investigate the contribution of dried cherry on phenols (TPh) and flavonoids (Fl) contents and antioxidant activities of honey. Phenolic compounds in Serbian polyfloral (PH), linden (LH) and acacia (AH) honey and also in their mixtures with dried cherry, in 40% mass concentrations (PH40; LH40, AH40), were determined. In comparison to honey, TPh increased 2.25 times for LH40, 2.16 times for AH40 and 1.45 times for PH40, while Fl increased 2.81-fold for PH40, 1.21-fold for LH40 and 1.44-fold for AH40. Antioxidant activity was investigated with two assays, DPPH test and reducing power (RP), and expressed as EC50DPPH and RP0.5 values. The EC50DPPH values were: EC50PH40 = 1.16 mg/ml; EC50LH40= 1.42 mg/ml and EC50AH40= 1.69 mg/ml, while RP0.5 were: RP0.5PH40 = 15.05 mg/ml; RP0.5LH40 = 16.09 mg/ml and P0.5AH40 = 17.60 mg/ml. Our results indicate that supplementation of polyfloral, linden and acacia honey with 40% dried cherry improves antioxidant activity of honey by enriching the phenolic composition.

Keywords: antioxidant activity, dried cherry, honey, phenolics

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Authors: Lorico D. S. Lapitan Jr., Yihan Xu, Yuan Guo, Dejian Zhou

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The ability of ultrasensitive detection of specific genes and discrimination of single nucleotide polymorphisms is important for clinical diagnosis and biomedical research. Herein, we report the development of a new ultrasensitive approach for label-free DNA detection using magnetic nanoparticle (MNP) assisted rapid target capture/separation in combination with signal amplification using poly-enzyme tagged polymer nanobead. The sensor uses an MNP linked capture DNA and a biotin modified signal DNA to sandwich bind the target followed by ligation to provide high single-nucleotide polymorphism discrimination. Only the presence of a perfect match target DNA yields a covalent linkage between the capture and signal DNAs for subsequent conjugation of a neutravidin-modified horseradish peroxidase (HRP) enzyme through the strong biotin-nuetravidin interaction. This converts each captured DNA target into an HRP which can convert millions of copies of a non-fluorescent substrate (amplex red) to a highly fluorescent product (resorufin), for great signal amplification. The use of polymer nanobead each tagged with thousands of copies of HRPs as the signal amplifier greatly improves the signal amplification power, leading to greatly improved sensitivity. We show our biosensing approach can specifically detect an unlabeled DNA target down to 10 aM with a wide dynamic range of 5 orders of magnitude (from 0.001 fM to 100.0 fM). Furthermore, our approach has a high discrimination between a perfectly matched gene and its cancer-related single-base mismatch targets (SNPs): It can positively detect the perfect match DNA target even in the presence of 100 fold excess of co-existing SNPs. This sensing approach also works robustly in clinical relevant media (e.g. 10% human serum) and gives almost the same SNP discrimination ratio as that in clean buffers. Therefore, this ultrasensitive SNP biosensor appears to be well-suited for potential diagnostic applications of genetic diseases.

Keywords: DNA detection, polymer beads, signal amplification, single nucleotide polymorphisms

Procedia PDF Downloads 233

Authors: Alireza Naseri, Susan L. Holdt, Charlotte Jacobsen

Abstract:

In 2014, the global amount of carrageenan production was 60,000 ton with a value of US$ 626 million. From this number, it can be estimated that the total dried seaweed consumption for this production was at least 300,000 ton/year. The protein content of these types of seaweed is 5 – 25%. If just half of this total amount of protein could be extracted, 18,000 ton/year of a high-value protein product would be obtained. The overall aim of this study was to develop a technology that will ensure further utilization of the seaweed that is used only as raw materials for carrageenan production as single extraction at present. More specifically, proteins should be extracted from the seaweed either before or after extraction of carrageenan with focus on maintaining the quality of carrageenan as a main product. Different mechanical, chemical and enzymatic technologies were evaluated. The optimized process was implemented in lab scale and based on its results; the new experiments were done a pilot and larger scale. In order to calculate the efficiency of the new upstream multi-extraction process, protein content was tested before and after extraction. After this step, the extraction of carrageenan was done and carrageenan content and the effect of extraction on yield were evaluated. The functionality and quality of carrageenan were measured based on rheological parameters. The results showed that by using the new multi-extraction process (submitted patent); it is possible to extract almost 50% of total protein without any negative impact on the carrageenan quality. Moreover, compared to the routine carrageenan extraction process, the new multi-extraction process could increase the yield of carrageenan and the rheological properties such as gel strength in the final carrageenan had a promising improvement. The extracted protein has initially been screened as a plant protein source in typical food applications. Further work will be carried out in order to improve properties such as color, solubility, and taste.

Keywords: carrageenan, extraction, protein, seaweed

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Authors: Rachel Fanelwa Ajayi, Anovuyo Jonnas, Emmanuel I. Iwuoha

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Tuberculosis (TB) is an infectious disease caused by the bacterium (Mycobacterium tuberculosis) which has a predilection for lung tissue due to its rich oxygen supply. The mycobacterial cell has a unique innate characteristic which allows it to resist human immune systems and drug treatments; hence, it is one of the most difficult of all bacterial infections to treat, let alone to cure. At the same time, multi-drug resistance TB (MDR-TB) caused by poorly managed TB treatment, is a growing problem and requires the administration of expensive and less effective second line drugs which take much longer treatment duration than fist line drugs. Therefore, to acknowledge the issues of patients falling ill as a result of inappropriate dosing of treatment and inadequate treatment administration, a device with a fast response time coupled with enhanced performance and increased sensitivity is essential. This study involved the synthesis of electroactive platforms for application in the development of nano-biosensors suitable for the appropriate dosing of clinically diagnosed patients by promptly quantifying the levels of the TB drug; Isonaizid. These nano-biosensors systems were developed on gold surfaces using the enzyme N-acetyletransferase 2 coupled to the cysteamine modified poly(8-anilino-1-napthalene sulphonic acid)/zinc oxide nanocomposites. The morphology of ZnO nanoparticles, PANSA/ZnO nano-composite and nano-biosensors platforms were characterized using High-Resolution Transmission Electron Microscopy (HRTEM) and High-Resolution Scanning Electron Microscopy (HRSEM). On the other hand, the elemental composition of the developed nanocomposites and nano-biosensors were studied using Fourier Transform Infra-Red Spectroscopy (FTIR) and Energy Dispersive X-Ray (EDX). The electrochemical studies showed an increase in electron conductivity for the PANSA/ZnO nanocomposite which was an indication that it was suitable as a platform towards biosensor development.

Keywords: N-acetyletransferase 2, isonaizid, tuberculosis, zinc oxide

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Authors: Nusrat Praween, Krishna Thej Pammi Guru, Palash Kumar Basu

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Designing non-invasive biosensors for cancer diagnosis is essential for developing an affordable and specific tool to measure cancer-related exosome biomarkers. Exosomes, released by healthy as well as cancer cells, contain valuable information about the biomarkers of various diseases, including cancer. Despite the availability of various isolation techniques, ultracentrifugation is the standard technique that is being employed. Post isolation, exosomes are traditionally exposed to detergents for extracting their proteins, which can often lead to protein degradation. Further to this, it is very essential to develop a sensing platform for the quantification of clinically relevant proteins in a wider range to ensure practicality. In this study, exosomes were immobilized on the Au Screen Printed Electrode (SPE) using EDC/NHS chemistry to facilitate binding. After immobilizing the exosomes on the screen-printed electrode (SPE), we investigated the impact of the electric field by applying various voltages to induce exosome lysis and release their contents. The lysed solution was used for sensing TSG101, a crucial biomarker associated with various cancers, using both faradaic and non-faradaic electrochemical impedance spectroscopy (EIS) methods. The results of non-faradaic and faradaic EIS were comparable and showed good consistency, indicating that non-faradaic sensing can be a reliable alternative. Hence, the non-faradaic sensing technique was used for label-free quantification of the TSG101 biomarker. The results were validated using ELISA. Our electrochemical immunosensor demonstrated a consistent response of TSG101 from 125 pg/mL to 8000 pg/mL, with a detection limit of 0.125 pg/mL at room temperature. Additionally, since non-faradic sensing is label-free, the ease of usage and cost of the final sensor developed can be reduced. The proposed immunosensor is capable of detecting the TSG101 protein at low levels in healthy serum with good sensitivity and specificity, making it a promising platform for biomarker detection.

Keywords: biosensor, exosomes isolation on SPE, electric field lysis of exosome, EIS sensing of TSG101

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Authors: Bello Gonzalez, Setten Van M., Brouwer M.

Abstract:

The chicken small intestine represents a dynamic and complex organ in which the enzymatic digestion and absorption of nutrients take place. The development of an in vitro fermentation chicken small intestinal model could be used as an alternative to explore the interaction between the microbiota and nutrient metabolism and to enhance the efficacy of targeting interventions to improve animal health. In the present study we have developed an in vitro fermentation chicken ileum microbiota model for unrevealing the complex interaction of ileum microbial community under physiological conditions. A two-vessel continuous fermentation process simulating in real-time the physiological conditions of the ileum content (pH, temperature, microaerophilic/anoxic conditions, and peristaltic movements) has been standardized as a proof of concept. As inoculum, we use a pool of ileum microbial community obtained from chicken broilers at the age of day 14. The development and validation of the model provide insight into the initial characterization of the ileum microbial community and its dynamics over time-related to nutrient assimilation and fermentation. Samples can be collected at different time points and can be used to determine the microbial compositional structure, dynamics, and diversity over time. The results of studies using this in vitro model will serve as the foundation for the development of a whole small intestine in vitro fermentation chicken gastrointestinal model to complement our already established in vitro fermentation chicken caeca model. The insight gained from this model could provide us with some information about the nutritional strategies to restore and maintain chicken gut homeostasis. Moreover, the in vitro fermentation model will also allow us to study relationships between gut microbiota composition and its dynamics over time associated with nutrients, antimicrobial compounds, and disease modelling.

Keywords: broilers, in vitro model, ileum microbiota, fermentation

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Authors: Si Yin Tee

Abstract:

Bimetallic nanostructures have received tremendous interests as a new class of nanomaterials which may have better technological usefulness with distinct properties from those of individual atoms and molecules or bulk matter. They excelled over the monometallic counterparts because of their improved electronic, optical and catalytic performances. The properties and the applicability of these bimetallic nanostructures not only depend on their size and shape, but also on the composition and their fine structure. These bimetallic nanostructures are potential candidates for bio-applications such as biosensing, bioimaging, biodiagnostics, drug delivery, targeted therapeutics, and tissue engineering. Herein, gold-incorporated copper (Cu/Au) nanostructures were synthesized through the controlled disproportionation of Cu⁺-oleylamine complex at 220 ºC to form copper nanowires and the subsequent reaction with Au³⁺ at different temperatures of 140, 220 and 300 ºC. This is to achieve their synergistic effect through the combined use of the merits of low-cost transition and high-stability noble metals. Of these Cu/Au nanostructures, Cu/Au nanotubes display the best performance towards electrochemical non-enzymatic glucose sensing, originating from the high conductivity of gold and the high aspect ratio copper nanotubes with high surface area so as to optimise the electroactive sites and facilitate mass transport. In addition to high sensitivity and fast response, the Cu/Au nanotubes possess high selectivity against interferences from other potential interfering species and excellent reproducibility with long-term stability. By introducing gold into copper nanostructures at a low level of 3, 1 and 0.1 mol% relative to initial copper precursor, a significant electrocatalytic enhancement of the resulting bimetallic Cu/Au nanostructures starts to occur at 1 mol%. Overall, the present fabrication of stable Cu/Au nanostructures offers a promising low-cost platform for sensitive, selective, reproducible and reusable electrochemical sensing of glucose.

Keywords: bimetallic, electrochemical sensing, glucose oxidation, gold-incorporated copper nanostructures

Procedia PDF Downloads 498

Authors: Chengyu Dong, Shao-Yuan Leu

Abstract:

Cellulosic ethanol production from lignocellulosic biomass can reduce our reliance on fossil fuel, mitigate climate change, and stimulate rural economic development. The relative low ethanol production (60 g/L) limits the economic viable of lignocellulose-based biorefinery. The ethanol production can be increased up to 80 g/L by removing nearly all the non-cellulosic materials, while the capital of the pretreatment process increased significantly. In this study, a fed-batch prehydrolysis simultaneously saccharification and fermentation process (PSSF) was designed to converse the sulfite pretreated softwood (~30% residual lignin) to high concentrations of ethanol (80 g/L). The liquefaction time of hydrolysis process was shortened down to 24 h by employing the fed-batch strategy. Washing out the spent liquor with water could eliminate the inhibition of the pretreatment spent liquor. However, the ethanol yield of lignocellulose was reduced as the fermentable sugars were also lost during the process. Fed-batch prehydrolyzing the while slurry (i.e. liquid plus solid fraction) pretreated softwood for 24 h followed by simultaneously saccharification and fermentation process at 28 °C can generate 80 g/L ethanol production. Fed-batch strategy is very effectively to eliminate the “solid effect” of the high gravity saccharification, so concentrating the cellulose to nearly 90% by the pretreatment process is not a necessary step to get high ethanol production. Detoxification of the pretreatment spent liquor caused the loss of sugar and reduced the ethanol yield consequently. The tolerance of yeast to inhibitors was better at 28 °C, therefore, reducing the temperature of the following fermentation process is a simple and valid method to produce high ethanol production.

Keywords: cellulosic ethanol, sulfite pretreatment, Fed batch PSSF, temperature

Procedia PDF Downloads 349

Authors: H. Aldewachi, M. Hines, M. McCulloch, N. Woodroofe, P. Gardiner

Abstract:

DPP-IV is an enzyme whose expression is affected in a variety of diseases, therefore, has been identified as possible diagnostic or prognostic marker for various tumours, immunological, inflammatory, neuroendocrine, and viral diseases. Recently, DPP-IV enzyme has been identified as a novel target for type II diabetes treatment where the enzyme is involved. There is, therefore, a need to develop sensitive and specific methods that can be easily deployed for the screening of the enzyme either as a tool for drug screening or disease marker in biological samples. A variety of assays have been introduced for the determination of DPP-IV enzyme activity using chromogenic and fluorogenic substrates, nevertheless these assays either lack the required sensitivity especially in inhibited enzyme samples or displays low water solubility implying difficulty for use in vivo samples in addition to labour and time-consuming sample preparation. In this study, novel strategies based on exploiting the high extinction coefficient of gold nanoparticles (GNPs) are investigated in order to develop fast, specific and reliable enzymatic assay by investigating synthetic peptide sequences containing a DPP IV cleavage site and coupling them to GNPs. The DPP IV could be detected by colorimetric response of peptide capped GNPs (P-GNPS) that could be monitored by a UV-visible spectrophotometer or even naked eyes, and the detection limit could reach 0.01 unit/ml. The P-GNPs, when subjected to DPP IV, showed excellent selectivity compared to other proteins (thrombin and human serum albumin) , which led to prominent colour change. This provided a simple and effective colorimetric sensor for on-site and real-time detection of DPP IV.

Keywords: gold nanoparticles, synthetic peptides, colorimetric detection, DPP-IV enzyme

Procedia PDF Downloads 284

Authors: Konstans Ruseva, Kristina Ivanova, Katerina Todorova, Margarita Gabrashanska, Tzanko Tzanov, Elena Vassileva

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Zwitterionic polymers (ZP) are well-known with their ultralow biofouling. They are successfully competing with poly(ethylene glycols) (PEG), which are considered as the “golden standard” in this respect. These unique properties are attributed to their strong hydration capacity, defined by the dipole-dipole interactions, arising between the ZP pendant groups as well as to the dipoles interaction with water molecules. Beside, ZP are highly resistant to bacterial adhesion thus ensuring an excellent anti-biofilm formation ability. Moreover, ZP are able to respond upon external stimuli such as temperature, pH, salt concentration changes which in combination with their anti-biofouling effect render this type of polymers as materials with a high potential in biomedical applications. The present work is focused on the development of zwitterionic hydrogels for efficient treatment of highly exudating and hard-to-heal chronic wounds. To this purpose, two types of ZP networks with different crosslinking degree were synthesized - polysulfobetaine (PSB) and polycarboxybetaine (PCB) ones. They were characterized in terms of their physico-mechanical properties, e.g. microhardness, swelling ability, smart behaviour. Furthermore, the potential of ZP networks to resist biofilm formation towards Staphylococcus aureus and Escherichia coli was studied. Their ability to reduce the high levels of myeloperoxidase and metalloproteinase, two enzymes that are part of the chronic wounds enviroenment, was revealed. Moreover, the in vitro cytotoxic assessment of PSB and PCB networks along with their in vivo performance in rats was also studied to reveal their high biocompatibility.

Keywords: absorption properties, biocompatibility, enzymatic inhibition activity, wound healing, zwitterionic polymers

Procedia PDF Downloads 171