Search results for: small multidrug resistance (SMR) protein
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 9712

Search results for: small multidrug resistance (SMR) protein

8992 Chemical Synthesis of a cDNA and Its Expression Analysis

Authors: Salman Akrokayan

Abstract:

Synthetic cDNA (ScDNA) of granulocyte colony-stimulating factor (G-CSF) was constructed using a DNA synthesizer with the aim to increase its expression level. 5' end of the ScDNA of G-CSF coding region was modified by decreasing the GC content without altering the predicted amino acids sequence. The identity of the resulting protein from ScDNA was confirmed by the highly specific enzyme-linked immunosorbent assay. In conclusion, a synthetic G-CSF cDNA in combination with the recombinant DNA protocol offers a rapid and reliable strategy for synthesizing the target protein. However, the commercial utilization of this methodology requires rigorous validation and quality control.

Keywords: synthetic cDNA, recombinant G-CSF, cloning, gene expression

Procedia PDF Downloads 269
8991 A Small Signal Model for Resonant Tunneling Diode

Authors: Rania M. Abdallah, Ahmed A. S. Dessouki, Moustafa H. Aly

Abstract:

This paper has presented a new simple small signal model for a resonant tunnelling diode device. The resonant tunnelling diode equivalent circuit elements were calculated and the results led to good agreement between the calculated equivalent circuit elements and the measurement results.

Keywords: resonant tunnelling diode, small signal model, negative differential conductance, electronic engineering

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8990 Cost Effective and Efficient Feeding: A Way Forward for Sustainable and Profitable Aquaculture

Authors: Pawan Kumar Sharma, J. Stephan Sampath Kumar, S. Anand, Chandana B. L.

Abstract:

Protein is the major component for the success in culture of shrimp and fishes. Apparently, excess dietary protein is undesirable, as it not only enhances the production cost but also leads to water quality deterioration. A field survey was conducted with aqua farmers of Kerala, India, a leading state in coastal aquaculture, to assess the role of protein component in feed that can be efficiently and effectively managed for sustainable aquaculture. The study showed an average feed amount of 13.55 ± 2.16 tonnes per hectare was being used by the farmers of Kerala. The average feed cost percentage of Rs. 57.76 ± 13.46 /kg was invested for an average protein level of 36.26 % ± 0.082 in the feed and Rs.78.95 ± 3.086 per kilogram of feed was being paid by the farmers. Study revealed that replacement of fish meal and fish oil within shrimp aquafeeds with alternative protein, and lipid sources can only be achieved if changes are made in the basic shrimp culturing practices, such as closed farming system through water recycling or zero-water exchange, and by maximizing in-situ, floc and natural food production within the culture system. The upshot of such production systems is that imports of high-quality feed ingredients and aqua feeds can eventually be eliminated, and the utilization of locally available feed ingredients from agricultural by-products can be greatly improved and maximized. The promotion of closed shrimp production systems would also greatly reduce water use and increase shrimp production per unit area but would necessitate the continuous provision of electricity for aeration during production. Alternative energy sources such as solar power might be used, and resource poor farming communities should also explore wind energy for use. The study concluded that farm made feed and closed farming systems are essential for the sustainability and profitability of the aquaculture industry.

Keywords: aqua feeds, floc, fish meal, protein, zero-water exchange

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8989 A Plasmonic Mass Spectrometry Approach for Detection of Small Nutrients and Toxins

Authors: Haiyang Su, Kun Qian

Abstract:

We developed a novel plasmonic matrix assisted laser desorption/ionization mass spectrometry (MALDI MS) approach to detect small nutrients and toxin in complex biological emulsion samples. We used silver nanoshells (SiO₂@Ag) with optimized structures as matrices and achieved direct analysis of ~6 nL of human breast milk without any enrichment or separation. We performed identification and quantitation of small nutrients and toxins with limit-of-detection down to 0.4 pmol (for melamine) and reaction time shortened to minutes, superior to the conventional biochemical methods currently in use. Our approach contributed to the near-future application of MALDI MS in a broad field and personalized design of plasmonic materials for real case bio-analysis.

Keywords: plasmonic materials, laser desorption/ionization, mass spectrometry, small nutrients, toxins

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8988 Antimicrobial Resistance Patterns of Campylobacter from Pig and Cattle Carcasses in Poland

Authors: Renata Szewczyk, Beata Lachtara, Kinga Wieczorek, Jacek Osek

Abstract:

Campylobacter is recognized as the main cause of bacterial gastrointestinal infections in Europe. A main source of the pathogen is poultry and poultry meat; however, other animals like pigs and cattle can also be reservoirs of the bacteria. Human Campylobacter infections are often self-limiting but in some cases, macrolide and fluoroquinolones have to be used. The aim of this study was to determine antimicrobial resistance patterns (AMR) of Campylobacter isolated from pig and cattle carcasses. Between July 2009 and December 2015, 735 swabs from pig (n = 457) and cattle (n = 278) carcasses were collected at Polish slaughterhouses. All samples were tested for the presence of Campylobacter by ISO 10272-1 and confirmed to species level using PCR. The antimicrobial susceptibility of Campylobacter isolates was determined by a microbroth dilution method with six antimicrobials: gentamicin (GEN), streptomycin (STR), erythromycin (ERY), nalidixic acid (NAL), ciprofloxacin (CIP) and tetracycline (TET). It was found that 167 of 735 samples (22.7%) were contaminated with Campylobacter. The vast majority of them were of pig origin (134; 80.2%), whereas for cattle carcasses Campylobacter was less prevalent (33; 19.8%). Among positive samples C. coli was predominant species (123; 73.7%) and it was isolated mainly from pig carcasses. The remaining isolates were identified as C. jejuni (44; 26.3%). Antimicrobial susceptibility indicated that 22 out of 167 Campylobacter (13.2%) were sensitive to all antimicrobials used. Fourteen of them were C. jejuni (63.6%; pig, n = 6; cattle, n = 8) and 8 was C. coli (36.4%; pig, n = 4; cattle, n = 4). Most of the Campylobacter isolates (145; 86.8%) were resistant to one or more antimicrobials (C. coli, n = 115; C. jejuni, n = 30). Comparing the AMR for Campylobacter species it was found that the most common pattern for C. jejuni was CIP-NAL-TET (9; 30.0%), whereas CIP-NAL-STR-TET was predominant among C. coli (47; 40.9%). Multiresistance, defined as resistance to three or more classes of antimicrobials, was found in 57 C. coli strains, mostly obtained from pig (52 isolates). On the other hand, only one C. jejuni strain, isolated from cattle, showed multiresistance with pattern CIP-NAL-STR-TET. Moreover, CIP-NAL-STR-TET was characteristic for most of multiresistant C. coli isolates (47; 82.5%). For the remaining C. coli the resistance patterns were CIP-ERY-NAL-TET (7 strains; 12.3%) and for one strain of each patterns: ERY-STR-TET, CIP-STR-TET, CIP-NAL-GEN-STR-TET. According to the present findings resistance to erythromycin was observed only in 11 C. coli (pig, n = 10; cattle, n = 1). In conclusion, the results of this study showed that pig carcasses may be a serious public health concern because of contamination with C. coli that might features multiresistance to antimicrobials.

Keywords: antimicrobial resistance, Campylobacter, carcasses, multi resistance

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8987 Experimental Investigation of Hull Form for Electric Driven Ferry

Authors: Vasilij Djackov, Tomas Zapnickas, Evgenii Iamshchikov, Lukas Norkevicius, Rima Mickeviciene, Larisa Vasiljeva

Abstract:

In this paper, the resistance and pitching values of the test of an electric ferry are presented. The research was carried out in the open flow channel of Klaipėda University with a multi-axis dynamometer. The received model resistance values were recalculated to the real vessel and the preliminary chosen propulsion unit power was compared. After analyzing the results of the pitching of the model, it was concluded that the shape of the hull needs to be further improved, taking into account the possible uneven weight distribution at the ends of the ferry. Further investigation of the hull of the electric ferry is recommended, including experiments with various water depths and activation of propulsion units.

Keywords: electrical ferry, model tests, open flow channel, pitching, resistance

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8986 Evaluation of Antimicrobial Efficacy of Nanofluid Containing Carbon Nanotubes Functionalized with Antibiotic on Urinary Tract Infection

Authors: Erfan Rahimi, Hadi Bahari Far, Mojgan Shikhpour

Abstract:

Background: Urinary tract infection is one of the most common nosocomial infections, especially among women. E. coli is one of the main causes of urinary tract infections and one of the most common antibiotics to fight this bacterium is ampicillin. As conventional antibiotics led to bacterial antibiotic resistance, modification of the pure drugs can address this issue. The aim of this study was to prepare nanofluids containing carbon nanotubes conjugated with ampicillin to improve drug performance and reduce antibiotic resistance. Methods: Multi-walled carbon nanotubes (MWCNTs) were activated with thionyl chloride by reflux system and nanofluids containing antibiotics were prepared by ultrasonic method. The properties of the prepared nano-drug were investigated by general element analysis, infrared spectroscopy, Raman spectroscopy, scanning electron microscopy and transmission electron microscopy. After the treatment of the desired strain with nanofluid, microbial studies were performed to evaluate the antibacterial effects and molecular studies were carried out to measure the expression of the resistance gene AcrAB. Result: We have shown that the antimicrobial effect of ampicillin-functionalized MWCNTs at low concentrations performed better than that of the conventional drug in both resistant and ATCC strains. Also, a decrease in antibiotic resistance of bacteria treated with ampicillin-functionalized MWCNTs compared to the pure drug was observed. Also, ampicillin-functionalized MWCNTs downregulated the expression of AcrAB in treated bacteria. Conclusion: Because carbon nanotubes are capable of destroying the bacterial wall, which provides antibiotic resistance features in bacteria, their usage in the form of nanofluids can make lower dosages (about three times less) than that of the pure drug more effective. Additionally, the expression of the bacterial resistance gene AcrAB decreased, thereby reducing antibiotic resistance and improving drug performance against bacteria.

Keywords: urinary tract infection, antibiotic resistance, carbon nanotube, nanofluid

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8985 Prevalence and Genetic Determinant of Drug Resistant Tuberculosis among Patients Completing Intensive Phase of Treatment in a Tertiary Referral Center in Nigeria

Authors: Aminu Bashir Mohammad, Agwu Ezera, Abdulrazaq G. Habib, Garba Iliyasu

Abstract:

Background: Drug resistance tuberculosis (DR-TB) continues to be a challenge in developing countries with poor resources. Routine screening for primary DR-TB before commencing treatment is not done in public hospitals in Nigeria, even with the large body of evidence that shows a high prevalence of primary DR-TB. Data on drug resistance and its genetic determinant among follow up TB patients is lacking in Nigeria. Hence the aim of this study was to determine the prevalence and genetic determinant of drug resistance among follow up TB patients in a tertiary hospital in Nigeria. Methods: This was a cross-sectional laboratory-based study conducted on 384 sputum samples collected from consented follow-up tuberculosis patients. Standard microbiology methods (Zeil-Nielsen staining and microscopy) and PCR (Line Probe Assay)] were used to analyze the samples collected. Person’s Chi-square was used to analyze the data generated. Results: Out of three hundred and eighty-four (384) sputum samples analyzed for mycobacterium tuberculosis (MTB) and DR-TB twenty-five 25 (6.5%) were found to be AFB positive. These samples were subjected to PCR (Line Probe Assay) out of which 18(72%) tested positive for DR-TB. Mutations conferring resistance to rifampicin (rpo B) and isoniazid (katG, and or inhA) were detected in 12/18(66.7%) and 6/18(33.3%), respectively. Transmission dynamic of DR-TB was not significantly (p>0.05) dependent on demographic characteristics. Conclusion: There is a need to strengthened the laboratory capacity for diagnosis of TB and drug resistance testing and make these services available, affordable, and accessible to the patients who need them.

Keywords: drug resistance tuberculosis, genetic determinant, intensive phase, Nigeria

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8984 Impact of pH Control on Peptide Profile and Antigenicity of Whey Hydrolysates

Authors: Natalia Caldeira De Carvalho, Tassia Batista Pessato, Luis Gustavo R. Fernandes, Ricardo L. Zollner, Flavia Maria Netto

Abstract:

Protein hydrolysates are ingredients of enteral diets and hypoallergenic formulas. Enzymatic hydrolysis is the most commonly used method for reducing the antigenicity of milk protein. The antigenicity and physicochemical characteristics of the protein hydrolysates depend on the reaction parameters. Among them, pH has been pointed out as of the major importance. Hydrolysis reaction in laboratory scale is commonly carried out under controlled pH (pH-stat). However, from the industrial point of view, controlling pH during hydrolysis reaction may be infeasible. This study evaluated the impact of pH control on the physicochemical properties and antigenicity of the hydrolysates of whey proteins with Alcalase. Whey protein isolate (WPI) solutions containing 3 and 7 % protein (w/v) were hydrolyzed with Alcalase 50 and 100 U g-1 protein at 60°C for 180 min. The reactions were carried out under controlled and uncontrolled pH conditions. Hydrolyses performed under controlled pH (pH-stat) were initially adjusted and maintained at pH 8.5. Hydrolyses carried out without pH control were initially adjusted to pH 8.5. Degree of hydrolysis (DH) was determined by OPA method, peptides profile was evaluated by HPLC-RP, and molecular mass distribution by SDS-PAGE/Tricine. The residual α-lactalbumin (α-La) and β-lactoglobulin (β-Lg) concentrations were determined using commercial ELISA kits. The specific IgE and IgG binding capacity of hydrolysates was evaluated by ELISA technique, using polyclonal antibodies obtained by immunization of female BALB/c mice with α-La, β-Lg and BSA. In hydrolysis under uncontrolled pH, the pH dropped from 8.5 to 7.0 during the first 15 min, remaining constant throughout the process. No significant difference was observed between the DH of the hydrolysates obtained under controlled and uncontrolled pH conditions. Although all hydrolysates showed hydrophilic character and low molecular mass peptides, hydrolysates obtained with and without pH control exhibited different chromatographic profiles. Hydrolysis under uncontrolled pH released, predominantly, peptides between 3.5 and 6.5 kDa, while hydrolysis under controlled pH released peptides smaller than 3.5 kDa. Hydrolysis with Alcalase under all conditions studied decreased by 99.9% the α-La and β-Lg concentrations in the hydrolysates detected by commercial kits. In general, β-Lg concentrations detected in the hydrolysates obtained under uncontrolled pH were significantly higher (p<0.05) than those detected in hydrolysates produced with pH control. The anti-α-La and anti-β-Lg IgE and IgG responses to all hydrolysates decreased significantly compared to WPI. Levels of specific IgE and IgG to the hydrolysates were below 25 and 12 ng ml-1, respectively. Despite the differences in peptide composition and α-La and β-Lg concentrations, no significant difference was found between IgE and IgG binding capacity of hydrolysates obtained with or without pH control. These results highlight the impact of pH on the hydrolysates characteristics and their concentrations of antigenic protein. Divergence between the antigen detection by commercial ELISA kits and specific IgE and IgG binding response was found in this study. This result shows that lower protein detection does not imply in lower protein antigenicity. Thus, the use of commercial kits for allergen contamination analysis should be cautious.

Keywords: allergy, enzymatic hydrolysis, milk protein, pH conditions, physicochemical characteristics

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8983 Marker Assisted Selection of Rice Genotypes for Xa5 and Xa13 Bacterial Leaf Blight Resistance Genes

Authors: P. Sindhumole, K. Soumya, R. Renjimol

Abstract:

Rice (Oryza sativa L.) is the major staple food crop over the world. It is prone to a number of biotic and abiotic stresses, out of which Bacterial Leaf Blight (BLB), caused by Xanthomonas oryzae pv. oryzae, is the most rampant. Management of this disease through chemicals or any other means is very difficult. The best way to control BLB is by the development of Host Plant Resistance. BLB resistance is not an activity of a single gene but it involves a cluster of more than thirty genes reported. Among these, Xa5 and Xa13 genes are two important ones, which can be diagnosed through marker assisted selection using closely linked molecular markers. During 2014, the first phase of field screening using forty traditional rice genotypes was carried out and twenty resistant symptomless genotypes were identified. Molecular characterisation of these genotypes using RM 122 SSR marker revealed the presence of Xa5 gene in thirteen genotypes. Forty-two traditional rice genotypes were used for the second phase of field screening for BLB resistance. Among these, sixteen resistant genotypes were identified. These genotypes, along with two susceptible check genotypes, were subjected to marker assisted selection for Xa13 gene, using the linked STS marker RG-136. During this process, presence of Xa13 gene could be detected in ten resistant genotypes. In future, these selected genotypes can be directly utilised as donors in Marker assisted breeding programmes for BLB resistance in rice.

Keywords: oryza sativa, SSR, STS, marker, disease, breeding

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8982 Assessment of Green Fluorescent Protein Signal for Effective Monitoring of Recombinant Fermentation Processes

Authors: I. Sani, A. Abdulhamid, F. Bello, Isah M. Fakai

Abstract:

This research has focused on the application of green fluorescent protein (GFP) as a new technique for direct monitoring of fermentation processes involving cultured bacteria. To use GFP as a sensor for pH and oxygen, percentage ratio of red fluorescence to green (% R/G) was evaluated. Assessing the magnitude of the % R/G ratio in relation to low or high pH and oxygen concentration, the bacterial strains were cultivated under aerobic and anaerobic conditions. SCC1 strains of E. coli were grown in a 5 L laboratory fermenter, and during the fermentation, the pH and temperature were controlled at 7.0 and 370C respectively. Dissolved oxygen tension (DOT) was controlled between 15-100% by changing the agitation speed between 20-500 rpm respectively. Effect of reducing the DOT level from 100% to 15% was observed after 4.5 h fermentation. There was a growth arrest as indicated by the decrease in the OD650 at this time (4.5-5 h). The relative fluorescence (green) intensity was decreased from about 460 to 420 RFU. However, %R/G ratio was significantly increased from about 0.1% to about 0.25% when the DOT level was decreased to 15%. But when the DOT was changed to 100%, a little increase in the RF and decrease in the %R/G ratio were observed. Therefore, GFP can effectively detect and indicate any change in pH and oxygen level during fermentation processes.

Keywords: Escherichia coli SCC1, fermentation process, green fluorescent protein, red fluorescence

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8981 Two-Protein Modified Gold Nanoparticles for Serological Diagnosis of Borreliosis

Authors: Mohammed Alasel, Michael Keusgen

Abstract:

Gold is a noble metal; in its nano-scale level (e.g. spherical nanoparticles), the conduction electrons are triggered to collectively oscillate with a resonant frequency when certain wavelengths of electromagnetic radiation interact with its surface; this phenomenon is known as surface plasmon resonance (SPR). SPR is responsible for giving the gold nanoparticles its intense red color depending mainly on its size, shape and distance between nanoparticles. A decreased distance between gold nanoparticles results in aggregation of them causing a change in color from red to blue. This aggregation enables gold nanoparticles to serve as a sensitive biosensoric indicator. In the proposed work, gold nanoparticles were modified with two proteins: i) Borrelia antigen, variable lipoprotein surface-exposed protein (VlsE), and ii) protein A. VlsE antigen induces a strong antibody response against Lyme disease and can be detected from early to late phase during the disease in humans infected with Borrelia. In addition, it shows low cross-reaction with the other non-pathogenic Borrelia strains. The high specificity of VlsE antigen to anti-Borrelia antibodies, combined simultaneously with the high specificity of protein A to the Fc region of all IgG human antibodies, was utilized to develop a rapid test for serological point of care diagnosis of borreliosis in human serum. Only in the presence of anti-Borrelia antibodies in the serum probe, an aggregation of gold nanoparticles can be observed, which is visible by a concentration-dependent colour shift from red (low IgG) to blue (high IgG). Experiments showed it is clearly possible to distinguish between positive and negative sera samples using a simple suspension of the two-protein modified gold nanoparticles in a very short time (30 minutes). The proposed work showed the potential of using such modified gold nanoparticles generally for serological diagnosis. Improved specificity and reduced assay time can be archived in applying increased salt concentrations combined with decreased pH values (pH 5).

Keywords: gold nanoparticles, gold aggregation, serological diagnosis, protein A, lyme borreliosis

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8980 The Behaviour of Laterally Loaded Piles Installed in the Sand with Enlarged Bases

Authors: J. Omer, H. Haroglu

Abstract:

Base enlargement in piles was invented to enhance pile resistance in downward loading, but the contribution of an enlarged base to the lateral load resistance of a pile has not been fully exploited or understood. This paper presents a laboratory investigation of the lateral capacity and deformation response of small-scale steel piles with enlarged bases installed in dry sand. Static loading tests were performed on 24 model piles having different base-to-shaft diameter ratios. The piles were installed in a box filled with dry sand, and lateral loads were applied to the pile tops using a pulley system. The test piles had shaft diameters of 20 mm, 16 mm, and 10 mm; base diameters of 900 mm, 700 mm, and 500 mm. As a control, a pile without base enlargement was tested to allow comparisons with the enlarged base piles. Incremental maintained loads were applied until pile failure approached while recording pile head deflections with high-precision dial gauges. The results showed that the lateral capacity increased with an increase in base diameter, albeit by different percentages depending on the shaft diameters and embedment length in the sand. There was always an increase in lateral capacity with increasing embedment length. Also, it was observed that an enlarged pile base had deflected less at a given load when compared to the control pile. Therefore, the research demonstrated the benefits of lateral capacity and stability of enlarging a pile base.

Keywords: pile foundations, enlarged base, lateral loading

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8979 Utilization of Rice and Corn Bran with Dairy By-Product in Tarhana Production

Authors: Kübra Aktaş, Nihat Akin

Abstract:

Tarhana is a traditional Turkish fermented food. It is widely consumed as soup and includes many different ingredients such as wheat flour, various vegetables, and spices, yoghurt, bakery yeast. It can also be enriched by adding other ingredients. Thus, its nutritional properties can be enhanced. In this study, tarhana was supplemented with two different types of brans (rice bran and corn bran) and WPC (whey protein concentrate powder) to improve its nutritional and functional properties. Some chemical properties of tarhana containing two different brans and their levels (0, 5, 10 and 15%) and WPC (0, 5, 10%) were investigated. The results indicated that addition of WPC increased ash content in tarhanas which were fortified with rice and corn bran. The highest antioxidant and phenolic content values were obtained with addition of rice bran in tarhana formulation. Compared to tarhana with corn bran, rice bran addition gave higher oil content values. The cellulose content of tarhana samples was determined between 0.75% and 2.74% and corn bran showed an improving effect on cellulose contents of samples. In terms of protein content, addition of WPC into the tarhana raised protein content for the samples.

Keywords: corn, rice, tarhana, whey

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8978 Molecular Farming: Plants Producing Vaccine and Diagnostic Reagent

Authors: Katerina H. Takova, Ivan N. Minkov, Gergana G. Zahmanova

Abstract:

Molecular farming is the production of recombinant proteins in plants with the aim to use the protein as a purified product, crude extract or directly in the planta. Plants gain more attention as expression systems compared to other ones due to the cost effective production of pharmaceutically important proteins, appropriate post-translational modifications, assembly of complex proteins, absence of human pathogens to name a few. In addition, transient expression in plant leaves enables production of recombinant proteins within few weeks. Hepatitis E virus (HEV) is a causative agent of acute hepatitis. HEV causes epidemics in developing countries and is primarily transmitted through the fecal-oral route. Presently, all efforts for development of Hepatitis E vaccine are focused on the Open Read Frame 2 (ORF2) capsid protein as it contains epitopes that can induce neutralizing antibodies. For our purpose, we used the CMPV-based vector-pEAQ-HT for transient expression of HEV ORF2 in Nicotiana benthamina. Different molecular analysis (Western blot and ELISA) showed that HEV ORF2 capsid protein was expressed in plant tissue in high-yield up to 1g/kg of fresh leaf tissue. Electron microscopy showed that the capsid protein spontaneously assembled in low abundance virus-like particles (VLPs), which are highly immunogenic structures and suitable for vaccine development. The expressed protein was recognized by both human and swine HEV positive sera and can be used as a diagnostic reagent for the detection of HEV infection. Production of HEV capsid protein in plants is a promising technology for further HEV vaccine investigations. Here, we reported for a rapid high-yield transient expression of a recombinant protein in plants suitable for vaccine production as well as a diagnostic reagent. Acknowledgments -The authors’ research on HEV is supported with grants from the Project PlantaSYST under the Widening Program, H2020 as well as under the UK Biotechnological and Biological Sciences Research Council (BBSRC) Institute Strategic Programme Grant ‘Understanding and Exploiting Plant and Microbial Secondary Metabolism’ (BB/J004596/1). The authors want to thank Prof. George Lomonossoff (JIC, Norwich, UK) for his contribution.

Keywords: hepatitis E virus, plant molecular farming, transient expression, vaccines

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8977 The Phosphatidate Phosphatase Pah1 and Its Regulator Nem1/spo7 Protein Phosphatase Required for Nucleophagy

Authors: Muhammad Arifur Rahman, Talukdar M. Waliullah, Takashi Ushimaru

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Nucleophagy selectively degrades nuclear materials, especially nucleolus after nutrient starvation or inactivation of TORC1 kinase in budding yeast. Budding yeast phosphatidate (PA) phosphatase Pah1 that converts PA to diacylglycerol is essential for partitioning of lipid precursors between membrane and storage that is crucial for many aspects of cell growth and development. Pah1 is required for nuclear/ER membrane biogenesis and vacuole function, but whether Pah1 and its activator Nem1/Spo7 protein phosphatase complex are involved in autophagy is largely unknown. Loss of Pah1 causes expansion of the nucleus and fragmentation of the vacuole. Here we show that Pah1 is required for bulk autophagy and nucleophagy after TORC1 inactivation. Loss of Pah1 impaired nucleophagy severely and bulk autophagy to a lesser extent. Loss of the Pah1 activator Nem1-Spo7 protein phosphatase exhibited similar features.

Keywords: autophagy, Nem1/Spo7 phosphatase, Pah1, nucleophagy, TORC1

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8976 Antimutagenic Activity of a Protein, Lectin Fraction from Urtica Dioica L.

Authors: Nijole Savickiene, Antonella Di Sotto, Gabriela Mazzanti, Rasa Starselskyte, Silvia Di Giacomo, Annabella Vitalone

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Plant lectins are non-enzymic and non-immune origin proteins that specifically recognize and bind to various sugar structures and possess the activity to agglutinate cells and/or precipitate polysaccharides and glycoconjugates. The emerging evidences showed that plant lectins contribute not only to tumour cell recognition but also to cell adhesion and localization, to signal transduction, to mitogenic cytotoxicity and apoptosis. Among chitin-binding lectins, the Urtica dioica agglutinin (UDA), which is a complex of different isoforms, has been poorly studied for its biological activity. In this context and according to the increasing interest for lectins as novel antitumor drugs, present paper aimed at evaluating the potential antimutagenic activity of a lectin-like glycoprotein-enriched fraction from aerial part of Urtica dioica L. Aim: to evaluate the potential chemopreventive properties of a protein - lectin fraction from the aerial part of Urtica dioica. Materials and methods: Protein – lectin fraction has been tested for the antimutagenic activity in bacteria (50–800 mg/plate; Ames test by the preincubation method) and for the cytotoxicity on human hepatoma HepG2 cells (0.06–2 mg/mL; 24 and 48 h incubation). Results: Protein – lectin fraction from stinging nettle was not cytotoxic on HepG2 cells up to 2 mg/mL; conversely, it exhibited a strong antimutagenic activity against the mutagen 2-aminoanthracene (2AA) in all strains tested (maximum inhibition of 56.78 and 61% in TA98, TA100, and WP2uvrA strains, respectively, at 800 mg/plate). Discussion and conclusions: Protein – lectin fraction from Urtica dioica L. possesses antimutagenic and radical scavenging properties. Being 2AA a pro-carcinogenic agent, we hypothesize that the antimutagenicity of it can be due to the inhibition of CYP450-isoenzymes, involved in the mutagen bioactivation.

Keywords: lectins, antimutagenicity, chemoprevention, Urtica dioica

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8975 Adaptation Mechanism and Planning Response to Resiliency Shrinking of Small Towns Based on Complex Adaptive System by Taking Wuhan as an Example

Authors: Yanqun Li, Hong Geng

Abstract:

The rapid urbanization process taking big cities as the main body leads to the unequal configuration of urban and rural areas in the aspects of land supply, industrial division of labor, service supply and space allocation, and induces the shrinking characterization of service energy, industrial system and population vitality in small towns. As an important spatial unit in the spectrum of urbanization that serves, connects and couples urban and rural areas, the shrinking phenomenon faced by small towns has an important influence on the healthy development of urbanization. Based on the census of small towns in Wuhan metropolitan area, we have found that the shrinking of small towns is a passive contraction of elastic tension under the squeeze in cities. Once affected by the external forces such as policy regulation, planning guidance, and population return, small towns will achieve expansion and growth. Based on the theory of complex adaptive systems, this paper comprehensively constructs the development index evaluation system of small towns from five aspects of population, economy, space, society and ecology, measures the shrinking level of small towns, further analyzes the shrinking characteristics of small towns, and identifies whether the shrinking is elastic or not. And then this paper measures the resilience ability index of small town contract from the above-mentioned five aspects. Finally, this paper proposes an adaptive mechanism of urban-rural interaction evolution under fine division of labor to response the passive shrinking in small towns of Wuhan. Based on the above, the paper creatively puts forward the planning response measures of the small towns on the aspects of spatial layout, function orientation and service support, which can provide reference for other regions.

Keywords: complex adaptive systems, resiliency shrinking, adaptation mechanism, planning response

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8974 Resistance Spot Welding of Boron Steel 22MnB5 with Complex Welding Programs

Authors: Szymon Kowieski, Zygmunt Mikno

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The study involved the optimization of process parameters during resistance spot welding of Al-coated martensitic boron steel 22MnB5, applied in hot stamping, performed using a programme with a multiple current impulse mode and a programme with variable pressure force. The aim of this research work was to determine the possibilities of a growth in welded joint strength and to identify the expansion of a welding lobe. The process parameters were adjusted on the basis of welding process simulation and confronted with experimental data. 22MnB5 steel is known for its tendency to obtain high hardness values in weld nuggets, often leading to interfacial failures (observed in the study-related tests). In addition, during resistance spot welding, many production-related factors can affect process stability, e.g. welding lobe narrowing, and lead to the deterioration of quality. Resistance spot welding performed using the above-named welding programme featuring 3 levels of force made it possible to achieve 82% of welding lobe extension. Joints made using the multiple current impulse program, where the total welding time was below 1.4s, revealed a change in a peeling mode (to full plug) and an increase in weld tensile shear strength of 10%.

Keywords: 22MnB5, hot stamping, interfacial fracture, resistance spot welding, simulation, single lap joint, welding lobe

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8973 Novel Development on Orthopedic Prosthesis by Nanocrystalline Hydroxyapatite Nanocomposite Coated on 316 L Stainless Steel

Authors: Neriman Ozada, Ebrahim Karamian, Amirsalar Khandan, Sina Ghafoorpoor Yazdi

Abstract:

Natural hydroxyapatite, NHA, coatings on the surface of 316 L stainless steel implants has been widely employed in order to achieve better osteoconductivity. For coating, the plasma spraying method is generally used because they ensure adhesion between the coating and the 316 L stainless steel (SS) surface. Some compounds such as zircon (ZrSiO4) is employed as an additive in an attempt to improve HA’s mechanical properties such as wear resistance and hardness. In this study wear resistance has been carried out in different chemical compositions of coating. Therefore, nanocomposites based on NHA containing of 0 wt.%, 5 wt.%, 10 wt.%, and 15 wt.% of zircon were used as a coating on the SS implants. The samples consisted of NHA, derived from calf heated at 850 °C for 3 h. The composite mixture was coated on SS by plasma spray method. The results were estimated using the scanning electron microscopy (SEM), X-ray diffraction (XRD) techniques were utilized to characterize the shape and size of NHA powder. Disc wear test and Vickers hardness were utilized to characterize the coated nanocomposite samples. The prepared NHA powder had nano-scale morphological structure with the mean crystallite size of 30-50 nm in diameter. The wear resistance are almost 320, 380, 415, and 395 m/g and hardness are approximately 376, 391, 420, 410 VHN in ceramic composite materials containing ZrSiO4. The results have been shown that the best wear resistance and hardness occurred in the sample coated by NHA/ZrSiO4 containing of 10 wt.% of zircon.

Keywords: zircon, 316 L stainless steel, wear resistance, orthopedic applications, plasma spray

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8972 Analysis and Modeling of Graphene-Based Percolative Strain Sensor

Authors: Heming Yao

Abstract:

Graphene-based percolative strain gauges could find applications in many places such as touch panels, artificial skins or human motion detection because of its advantages over conventional strain gauges such as flexibility and transparency. These strain gauges rely on a novel sensing mechanism that depends on strain-induced morphology changes. Once a compression or tension strain is applied to Graphene-based percolative strain gauges, the overlap area between neighboring flakes becomes smaller or larger, which is reflected by the considerable change of resistance. Tiny strain change on graphene-based percolative strain sensor can act as an important leverage to tremendously increase resistance of strain sensor, which equipped graphene-based percolative strain gauges with higher gauge factor. Despite ongoing research in the underlying sensing mechanism and the limits of sensitivity, neither suitable understanding has been obtained of what intrinsic factors play the key role in adjust gauge factor, nor explanation on how the strain gauge sensitivity can be enhanced, which is undoubtedly considerably meaningful and provides guideline to design novel and easy-produced strain sensor with high gauge factor. We here simulated the strain process by modeling graphene flakes and its percolative networks. We constructed the 3D resistance network by simulating overlapping process of graphene flakes and interconnecting tremendous number of resistance elements which were obtained by fractionizing each piece of graphene. With strain increasing, the overlapping graphenes was dislocated on new stretched simulation graphene flake simulation film and a new simulation resistance network was formed with smaller flake number density. By solving the resistance network, we can get the resistance of simulation film under different strain. Furthermore, by simulation on possible variable parameters, such as out-of-plane resistance, in-plane resistance, flake size, we obtained the changing tendency of gauge factor with all these variable parameters. Compared with the experimental data, we verified the feasibility of our model and analysis. The increase of out-of-plane resistance of graphene flake and the initial resistance of sensor, based on flake network, both improved gauge factor of sensor, while the smaller graphene flake size gave greater gauge factor. This work can not only serve as a guideline to improve the sensitivity and applicability of graphene-based strain sensors in the future, but also provides method to find the limitation of gauge factor for strain sensor based on graphene flake. Besides, our method can be easily transferred to predict gauge factor of strain sensor based on other nano-structured transparent optical conductors, such as nanowire and carbon nanotube, or of their hybrid with graphene flakes.

Keywords: graphene, gauge factor, percolative transport, strain sensor

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8971 Regulation on Macrophage and Insulin Resistance after Aerobic Exercise in High-Fat Diet Mice

Authors: Qiaofeng Guo

Abstract:

Aims: Obesity is often accompanied by insulin resistance (IR) and whole-body inflammation. Aerobic exercise is an effective treatment to improve insulin resistance and inflammation. However, the anti-inflammatory mechanisms of exercise on epididymal and subcutaneous adipose remain to be elucidated. Here, we compared the macrophage polarization between epididymal and subcutaneous adipose after aerobic exercise. Methods: Male C57BL/6 mice were fed a normal diet group or a high-fat diet group for 12 weeks and performed aerobic training on a treadmill at 55%~65% VO₂ max for eight weeks. Food intake, body weight, and fasting blood glucose levels were monitored weekly. The intraperitoneal glucose tolerance test was to evaluate the insulin resistance model. Fat mass, blood lipid profile, serum IL-1β, TNF-α levels, and CD31/CD206 rates were analysed after the intervention. Results: FBG (P<0.01), AUCIPGTT (P<0.01), and HOMA-IR (P<0.01) increased significantly for a high-fat diet and decreased significantly after the exercise. Eight weeks of aerobic exercise attenuated HFD-induced weight gain and glucose intolerance and improved insulin sensitivity. Serum IL-1β, TNF-α, CD11C/CD206 expression in subcutaneous adipose tissue were not changed before and after exercise, but not in epididymal adipose tissue (P<0.01). Conclusion: Insulin resistance is not accompanied by chronic inflammation and M1 polarization of subcutaneous adipose tissue macrophages in high-fat diet mice. Aerobic exercise effectively improved lipid metabolism and insulin sensitivity, which may be closely associated with reduced M1 polarization of epididymal adipose macrophages.

Keywords: aerobic exercise, insulin resistance, chronic inflammation, adipose, macrophage polarization

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8970 Small Text Extraction from Documents and Chart Images

Authors: Rominkumar Busa, Shahira K. C., Lijiya A.

Abstract:

Text recognition is an important area in computer vision which deals with detecting and recognising text from an image. The Optical Character Recognition (OCR) is a saturated area these days and with very good text recognition accuracy. However the same OCR methods when applied on text with small font sizes like the text data of chart images, the recognition rate is less than 30%. In this work, aims to extract small text in images using the deep learning model, CRNN with CTC loss. The text recognition accuracy is found to improve by applying image enhancement by super resolution prior to CRNN model. We also observe the text recognition rate further increases by 18% by applying the proposed method, which involves super resolution and character segmentation followed by CRNN with CTC loss. The efficiency of the proposed method shows that further pre-processing on chart image text and other small text images will improve the accuracy further, thereby helping text extraction from chart images.

Keywords: small text extraction, OCR, scene text recognition, CRNN

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8969 Study of Temperature Difference and Current Distribution in Parallel-Connected Cells at Low Temperature

Authors: Sara Kamalisiahroudi, Jun Huang, Zhe Li, Jianbo Zhang

Abstract:

Two types of commercial cylindrical lithium ion batteries (Panasonic 3.4 Ah NCR-18650B and Samsung 2.9 Ah INR-18650), were investigated experimentally. The capacities of these samples were individually measured using constant current-constant voltage (CC-CV) method at different ambient temperatures (-10 ℃, 0 ℃, 25 ℃). Their internal resistance was determined by electrochemical impedance spectroscopy (EIS) and pulse discharge methods. The cells with different configurations of parallel connection NCR-NCR, INR-INR and NCR-INR were charged/discharged at the aforementioned ambient temperatures. The results showed that the difference of internal resistance between cells much more evident at low temperatures. Furthermore, the parallel connection of NCR-NCR exhibits the most uniform temperature distribution in cells at -10 ℃, this feature is quite favorable for the safety of the battery pack.

Keywords: batteries in parallel connection, internal resistance, low temperature, temperature difference, current distribution

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8968 Stenotrophomonas maltophilia: The Major Carbapenem Resistance Bacteria from Waste Water Treatment Plant of Pig Farm

Authors: Young-Ji Kim, Jin-Hyeong Park, Hong-Seok Kim, Jung-Whan Chon, Kwang-Yeop Kim, Dong-Hyeon Kim, Il-Byeong Kang, Da-Na Jeong, Jin-Hyeok Yim, Ho-Seok Jang, Kwang-Young Song, Kun-Ho Seo

Abstract:

Stenotrophomonas maltophilia is one of the emerging opportunistic pathogens, and also known to have extensive drug resistance intrinsically including carbepenems which is last resort for most serious infections. One possible way for S. maltophilia to infect human is via wastewater treatment plant (WWTP). In the period between October 2016 and February 2017, effluent samples of WWTP from 3 different pig farms were collected once a month and screened for isolation of S. maltophilia. Total 16 strains of S. maltophilia were isolated and, the antibiotic susceptibility phenotypes were determined by Vitek 2 system for 16 antibiotics, ampicillin (AMP), amoxicillin/clavulanic acid (AMC), piperacillin/tazobactam (TZP), cefazolin (CZ), cefoxitin (FOX), cefotaxime (CTX), ceftazidime (CAZ), cefepime (FEP), aztreonam (AZT), ertapenem (ETP), imipenem (IMP), amikacin (AK), gentamicin (GN), ciprofloxacin (CIP), tigecycline (TGC) and trimethoprim/sulfamethoxazole (SXT). All isolates showed high resistance to AMP (100%), CZ (100%), FOX (100%), CTX (100%), CAZ (100%), FEP (94%), AZT (100%), ETP (100%), IMP (100%), AK (100%), GN (100%) whereas were susceptible to CIP (0%), TGC (0%), SXT (6%). All strains harbored at least one of the antibiotic resistance determinant such as spgM, rmlA, and rpfF. Some isolates had similar MLST (multilocus sequence typing) types with clinical isolates, suggesting WWTP could have potential role in the transmission of S. maltophilia to aquatic environment and, possibly, to humans.

Keywords: Stenotrophomonas maltophilia, Carbapenem resistance, waste water treatment plant, pig farm

Procedia PDF Downloads 452
8967 Comparison Between Vegans and Omnivores on the Recovery of Delayed Onset Muscle Soreness in Young Females

Authors: Njeim Pressila, Hajj-Boutros Guy, Antony D. Karelis

Abstract:

Background: Acute resistance exercise is associated with an elevated inflammation response, which could lead to delayed onset muscle soreness (DOMS). There is evidence that suggests that ingesting foods that have anti-inflammation properties may help reduce DOMS. A vegan diet has also been shown to be an anti-inflammatory diet which could, in turn, decrease DOMS. Objective and hypothesis: The purpose of the present study will be to compare markers of DOMS between vegans and omnivores after acute resistance exercise in young females. We hypothesize that vegans will have a better recovery of DOMS markers after a resistance exercise session compared to omnivores. Methods: Population: We will recruit30 vegans and 30 omnivores to participate in this study. Allvolunteers will follow either a vegan or an omnivore diet for at least 2 years. Anthropometric measurements, body composition, musclestrength (leg and chest press), markers of DOMS (swelling, pain, and stiffness), and dietary factors, as well as a wellness and anxiety questionnaire will be measured. All participants will also perform an acute resistance exercise session in order to induce DOMS. Pertinence: This project will give us a better understanding on the recovery process of vegans after a resistance training session and, as such, provide useful information to health professionals and athletes/coaches (kinesiologists and nutritionists)

Keywords: vgeans, omnivores, delayed onset muscle soreness, pain, stifness

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8966 Tribocorrosion Behavior of Austempered Ductile Iron Microalloyed with Boron

Authors: S. Gvazava, N. Khidasheli, G. Gordeziani, A. DL. Batako

Abstract:

The work presented in this paper studied the tribological characteristics (wear resistance, friction coefficient) of austempered ductile iron (ADI) with different combinations of structural composition (upper bainite, lower bainite, retained austenite) in dry sliding friction. A range of structural states of the metal matrix was obtained by changing the regimes of isothermal quenching of high-strength cast iron. The tribological tests were carried out using two sets of isothermal quenched cast irons. After austenitization at 900°С for 60 minutes, the specimens from the first group were isothermally quenched at the 300°С temperature and the specimens from the second set – at 400°С. The investigations showed that the isothermal quenching increases the friction coefficient of high-strength cast irons. The friction coefficient was found to be in the range from 0.4 to 0.55 for cast irons, depending on the structures of the metal matrix. The quenched cast irons having lower bainite demonstrate higher wear resistance in dry friction conditions. The dependence of wear resistance on the amount of retained austenite in isothermal quenched cast irons has a nonlinear characteristic and reaches its maximum value when the content of retained austenite is about 15-22%. The boron micro-additives allowed to reduce the friction coefficient of ADI and increase their wear resistance by 1.5-1.7 times.

Keywords: wear resistance, dry sliding, austempering, ADI, friction coefficient, retained austenite, isothermal quenching

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8965 Reversibility of Photosynthetic Activity and Pigment-protein Complexes Expression During Seed Development of Soybean and Black Soybean

Authors: Tzan-Chain Lee

Abstract:

Seeds are non-leaves green tissues. Photosynthesis begins with light absorption by chlorophyll and then the energy transfer between two pigment-protein complexes (PPC). Most studies of photosynthesis and PPC expression were focused on leaves; however, during seeds’ development were rare. Developed seeds from beginning pod (stage R3) to dried seed (stage R8), and the dried seed after sowing for 1-4 day, were analyzed for their chlorophyll contents. Thornber and MARS gel systems analysis compositions of PPC. Chlorophyll fluorescence was used to detect maximal photosynthetic efficiency (Fv/Fm). During soybean and black soybean seeds development (stages R3-R6), Fv/Fm up to 0.8, and then down-regulated after full seed (stage R7). In dried seed (stage R8), the two plant seeds lost photosynthetic activity (Fv/Fm=0), but chlorophyll degradation only occurred in soybean after full seed. After seeds sowing for 4 days, chlorophyll drastically increased in soybean seeds, and Fv/Fm recovered to 0.8 in the two seeds. In PPC, the two soybean seeds contained all PPC during seeds development (stages R3-R6), including CPI, CPII, A1, AB1, AB2, and AB3. However, many proteins A1, AB1, AB2, and CPI were totally missing in the two dried seeds (stage R8). The deficiency of these proteins in dried seeds might be caused by the incomplete photosynthetic activity. After seeds germination and seedling exposed to light for 4 days, all PPC were recovered, suggesting that completed PPC took place in the two soybean seeds. This study showed the reversibility of photosynthetic activity and pigment-protein complexes during soybean and black soybean seeds development.

Keywords: light-harvesting complex, pigment–protein complexes, soybean cotyledon, grana development

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8964 Functionalization of Carbon-Coated Iron Nanoparticles with Fluorescent Protein

Authors: A. G. Pershina, P. S. Postnikov, M. E. Trusova, D. O. Burlakova, A. E. Sazonov

Abstract:

Invention of magnetic-fluorescent nanocomposites is a rapidly developing area of research. The magnetic-fluorescent nanocomposite attractiveness is connected with the ability of simultaneous management and control of such nanocomposites by two independent methods based on different physical principles. These nanocomposites are applied for the solution of various essential scientific and experimental biomedical problems. The aim of this research is development of principle approach to nanobiohybrid structures with magnetic and fluorescent properties design. The surface of carbon-coated iron nanoparticles (Fe@C) were covalently modified by 4-carboxy benzenediazonium tosylate. Recombinant fluorescent protein TagGFP2 (Eurogen) was obtained in E. coli (Rosetta DE3) by standard laboratory techniques. Immobilization of TagGFP2 on the nanoparticles surface was provided by the carbodiimide activation. The amount of COOH-groups on the nanoparticle surface was estimated by elemental analysis (Elementar Vario Macro) and TGA-analysis (SDT Q600, TA Instruments. Obtained nanocomposites were analyzed by FTIR spectroscopy (Nicolet Thermo 5700) and fluorescence microscopy (AxioImager M1, Carl Zeiss). Amount of the protein immobilized on the modified nanoparticle surface was determined by fluorimetry (Cary Eclipse) and spectrophotometry (Unico 2800) with the help of preliminary obtained calibration plots. In the FTIR spectra of modified nanoparticles the adsorption band of –COOH group around 1700 cm-1 and bands in the region of 450-850 cm-1 caused by bending vibrations of benzene ring were observed. The calculated quantity of active groups on the surface was equal to 0,1 mmol/g of material. The carbodiimide activation of COOH-groups on nanoparticles surface results to covalent immobilization of TagGFP2 fluorescent protein (0.2 nmol/mg). The success of immobilization was proved by FTIR spectroscopy. Protein characteristic adsorption bands in the region of 1500-1600 cm-1 (amide I) were presented in the FTIR spectrum of nanocomposite. The fluorescence microscopy analysis shows that Fe@C-TagGFP2 nanocomposite possesses fluorescence properties. This fact confirms that TagGFP2 protein retains its conformation due to immobilization on nanoparticles surface. Magnetic-fluorescent nanocomposite was obtained as a result of unique design solution implementation – the fluorescent protein molecules were fixed to the surface of superparamagnetic carbon-coated iron nanoparticles using original diazonium salts.

Keywords: carbon-coated iron nanoparticles, diazonium salts, fluorescent protein, immobilization

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8963 Polymorphisms of STAT5A and DGAT1 Genes and Their Associations with Milk Trait in Egyptian Goats

Authors: Othman Elmahdy Othman

Abstract:

The objectives of this study were to identify polymorphisms in the STAT5A using Restriction Fragment Length Polymorphism and DGAT1 using Single-Strand Conformation Polymorphism genes among three Egyptian goat breeds (Barki, Zaraibi, and Damascus) as well as investigate the effect of their genotypes on milk composition traits of Zaraibi goats. One hundred and fifty blood samples were collected for DNA extraction, 60 from Zaraibi, 40 from Damascus and 50 from Barki breeds. Fat, protein and lactose percentages were determined in Zaraibi goat milk using an automatic milk analyzer. Two genotypes, CC and CT (for STAT5A) and C-C- and C-C+ (for DGAT1), were identified in the three Egyptian goat breeds with different frequencies. The associations between these genotypes and milk fat, protein and lactose were determined in Zaraibi breed. The results showed that the STAT5A genotypes had significant effects on milk yield, protein, fat and lactose with the superiority of CT genotype over CC. Regarding DGAT1 polymorphism, the result showed the only association between it with milk fat where the animals with C-C+ genotype had greater milk fat than animals possess C-C- genotype. The association of combined genotypes with milk trait declared that the does with heterozygous genotypes for both genes are preferred than does with homozygous genotypes where the animals with CTC-C+ have more milk yield, fat and protein than those with CCC-C- genotype. In conclusion, the result showed that C/T and C-/C+ SNPs of STAT5A and DGAT1 genes respectively may be useful markers for assisted selection programs to improve goat milk composition

Keywords: DGAT1, genetic polymorphism, milk trait, STAT5A

Procedia PDF Downloads 144