Search results for: monoclonal antibodies.

Authors: Ishan Arora, Anurag S. Rathore

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The primary objective of this work was to study the effect of resin chemistry, pH and molarity of binding and elution buffer on aggregate removal using Cation Exchange Chromatography and find the optimum conditions which can give efficient aggregate removal with minimum loss of yield. Four different resins were used for carrying out the experiments: Fractogel EMD SO3 -(S), Fractogel EMD COO-(M), Capto SP ImpRes and S Ceramic HyperD. Runs were carried out on the AKTA Avant system. Design of Experiments (DOE) was used for analysis using the JMP software. The dependence of the yield obtained using different resins on the operating conditions was studied. Success has been achieved in obtaining yield greater than 90% using Capto SP ImpRes and Fractogel EMD COO-(M) resins. It has also been found that a change in the operating conditions generally has different effects on the yields obtained using different resins.

Keywords: Aggregates, cation exchange chromatography, design of experiments, monoclonal antibodies.

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Authors: Simona Dostalova, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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To ensure targeting of apoferritin nanocarrier with encapsulated doxorubicin drug, we used a peptide linker based on a protein G with N-terminus affinity towards Fc region of antibodies. To connect the peptide to the surface of apoferritin, the C-terminus of peptide was made of cysteine with affinity to gold. The surface of apoferritin with encapsulated doxorubicin (APODOX) was coated either with gold nanoparticles (APODOX-Nano) or gold(III) chloride hydrate reduced with sodium borohydride (APODOX-HAu). The reduction with sodium borohydride caused a loss of doxorubicin fluorescent properties and probably accompanied with the loss of its biological activity. Fluorescent properties of APODOX-Nano were similar to the unmodified APODOX; therefore it was more suited for the intended use. To evaluate the specificity of apoferritin modified with antibodies, ELISA-like method was used with the surface of microtitration plate wells coated by the antigen (goat anti-human IgG antibodies). To these wells, the nanocarrier was applied. APODOX without the modification showed 5× lower affinity to the antigen than APODOX-Nano modified gold and targeting antibodies (human IgG antibodies).

Keywords: Antibody targeting, apoferritin, doxorubicin, nanocarrier.

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Authors: A. Asambe, A. K. B. Sackey, L. B. Tekdek

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A serosurveillance study was conducted to detect the presence of antibodies to African swine fever virus (ASFV) and Classical swine fever virus in pigs sampled from piggeries and Makurdi central slaughter slab in Benue State, Nigeria. 416 pigs from 74 piggeries across 12 LGAs and 44 pigs at the Makurdi central slaughter slab were sampled for serum. The sera collected were analysed using Indirect Enzyme Linked Immunosorbent Assay (ELISA) test kit to test for antibodies to ASFV, while competitive ELISA test kit was used to test for antibodies to CSFV. Of the 416 pigs from piggeries and 44 pigs sampled from the slaughter slab, seven (1.7%) and six (13.6%), respectively, tested positive to ASFV antibodies and was significantly associated (p < 0.0001). Out of the 12 LGAs sampled, Obi LGA had the highest ASFV antibody detection rate of (4.8%) and was significantly associated (p < 0.0001). None of the samples tested positive to CSFV antibodies. The study concluded that antibodies to CSFV were absent in the sampled pigs in piggeries and at the Makurdi central slaughter slab in Benue State, while antibodies to ASFV were present in both locations; hence, the need to keep an eye open for CSF too since both diseases may pose great risk in the study area. Further studies to characterise the ASFV circulating in Benue State and investigate the possible sources is recommended. Routine surveillance to provide a comprehensive and readily accessible data base to plan for the prevention of any fulminating outbreak is also recommended.

Keywords: African swine fever, classical swine fever, piggery, slaughter slab, surveillance.

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Authors: Maysam Mard-Soltani, Mohamad Javad Rasaee, Saeed Khalili, Abdol Karim Sheikhi, Mehdi Hedayati

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Production of specific antibody responses against hTSH is a cumbersome process due to the high identity between the hTSH and the other members of the glycoprotein hormone family (FSH, LH and HCG) and the high identity between the human hTSH and host animals for antibody production. Therefore, two polyclonal antibodies were purified against two recombinant proteins. Four possible ELISA tests were designed based on these antibodies. These ELISA tests were checked against hTSH and other glycoprotein hormones, and their sensitivity and specificity were assessed. Bioinformatics tools were used to analyze the immunological properties. After the immunogen region selection from hTSH protein, c terminal of B hTSH was selected and applied. Two recombinant genes, with these cut pieces (first: two repeats of C terminal of B hTSH, second: tetanous toxin+B hTSH C terminal), were designed and sub-cloned into the pET32a expression vector. Standard methods were used for protein expression, purification, and verification. Thereafter, immunizations of the white New Zealand rabbits were performed and the serums of them were used for antibody titration, purification and characterization. Then, four ELISA tests based on two antibodies were employed to assess the hTSH and other glycoprotein hormones. The results of these assessments were compared with standard amounts. The obtained results indicated that the desired antigens were successfully designed, sub-cloned, expressed, confirmed and used for in vivo immunization. The raised antibodies were capable of specific and sensitive hTSH detection, while the cross reactivity with the other members of the glycoprotein hormone family was minimum. Among the four designed tests, the test in which the antibody against first protein was used as capture antibody, and the antibody against second protein was used as detector antibody did not show any hook effect up to 50 miu/l. Both proteins have the ability to induce highly sensitive and specific antibody responses against the hTSH. One of the antibody combinations of these antibodies has the highest sensitivity and specificity in hTSH detection.

Keywords: hTSH, bioinformatics, protein expression, cross reactivity.

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Authors: Eszter Tóth, Ervin Welker

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Shadoo protein (Sho) was described in 2003 as the newest member of Prion protein superfamily [1]. Sho has similar structural motifs like prion protein (PrP) that is known for its central role in transmissible spongiform enchephalopathies. Although a great number of functions have been proposed, the exact physiological function of PrP is not known yet. Investigation of the function and localization of Sho may help us to understand the function of the Prion protein superfamily. Analyzing the subcellular localization of YFP-tagged forms of Sho, we detected the protein in the plasma membrane and in the nucleus of various cell lines. To reveal the localization of the endogenous protein we generated antibodies against Shadoo as well as employed commercially available anti-Shadoo antibodies: i) EG62 anti-mouse Shadoo antibody generated by Eurogentec Ltd.; ii) S-12 anti-human Shadoo antibody by Santa Cruz Biotechnology Inc.; iii) R-12 anti-mouse Shadoo antibody by Santa Cruz Biotechnology Inc.; iv) SPRN antibody against human Shadoo by Abgent Inc. We carried out immunocytochemistry on non-transfected HeLa, Zpl 2-1, Zw 3-5, GT1-1, GT1-7 and SHSY5Y cells as well as on YFP-Sho, Sho-YFP, and YFP-GPI transfected HeLa cells. Their specificity (in antibody-peptide competition assay) and co-localization (with the YFP signal) were assessed.

Keywords: Shadoo, prion protein, immunocytochemistry, antibody-peptide competition assay, antibody.

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Authors: Rujira Kongnuy, Puntani Pongsumpun, I-Ming Tang

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Mathematical models can be used to describe the dynamics of the spread of infectious disease between susceptibles and infectious populations. Dengue fever is a re-emerging disease in the tropical and subtropical regions of the world. Its incidence has increased fourfold since 1970 and outbreaks are now reported quite frequently from many parts of the world. In dengue endemic regions, more cases of dengue infection in pregnancy and infancy are being found due to the increasing incidence. It has been reported that dengue infection was vertically transmitted to the infants. Primary dengue infection is associated with mild to high fever, headache, muscle pain and skin rash. Immune response includes IgM antibodies produced by the 5th day of symptoms and persist for 30-60 days. IgG antibodies appear on the 14th day and persist for life. Secondary infections often result in high fever and in many cases with hemorrhagic events and circulatory failure. In the present paper, a mathematical model is proposed to simulate the succession of dengue disease transmission in pregnancy and infancy. Stability analysis of the equilibrium points is carried out and a simulation is given for the different sets of parameter. Moreover, the bifurcation diagrams of our model are discussed. The controlling of this disease in infant cases is introduced in the term of the threshold condition.

Keywords: Dengue infection, equilibrium states, maternalantibodies, pregnancy and infancy.

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Authors: Rujira Kongnuy, Puntani Pongsumpun, I-Ming Tang

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Dengue fever is an important human arboviral disease. Outbreaks are now reported quite often from many parts of the world. The number of cases involving pregnant women and infant cases are increasing every year. The illness is often severe and complications may occur. Deaths often occur because of the difficulties in early diagnosis and in the improper management of the diseases. Dengue antibodies from pregnant women are passed on to infants and this protects the infants from dengue infections. Antibodies from the mother are transferred to the fetus when it is still in the womb. In this study, we formulate a mathematical model to describe the transmission of this disease in pregnant women. The model is formulated by dividing the human population into pregnant women and non-pregnant human (men and non-pregnant women). Each class is subdivided into susceptible (S), infectious (I) and recovered (R) subclasses. We apply standard dynamical analysis to our model. Conditions for the local stability of the equilibrium points are given. The numerical simulations are shown. The bifurcation diagrams of our model are discussed. The control of this disease in pregnant women is discussed in terms of the threshold conditions.

Keywords: Dengue disease, local stability, mathematical model, pregnancy.

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Authors: Pei-Chann Chang, Wei-Hsiu Huang, Ching-Jung Ting, Hwei-Wen Luo, Yu-Peng Yu

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Artificial Immune System is applied as a Heuristic Algorithm for decades. Nevertheless, many of these applications took advantage of the benefit of this algorithm but seldom proposed approaches for enhancing the efficiency. In this paper, a Self-evolving Artificial Immune System is proposed via developing the T and B cell in Immune System and built a self-evolving mechanism for the complexities of different problems. In this research, it focuses on enhancing the efficiency of Clonal selection which is responsible for producing Affinities to resist the invading of Antigens. T and B cell are the main mechanisms for Clonal Selection to produce different combinations of Antibodies. Therefore, the development of T and B cell will influence the efficiency of Clonal Selection for searching better solution. Furthermore, for better cooperation of the two cells, a co-evolutional strategy is applied to coordinate for more effective productions of Antibodies. This work finally adopts Flow-shop scheduling instances in OR-library to validate the proposed algorithm.

Keywords: Artificial Immune System, Clonal Selection, Flow-shop Scheduling Problems, Co-evolutional strategy

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Authors: Natasha Hussain, Maleeha Aslam, Robina Farooq

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Hepatitis B and hepatitis C are among the most significant hepatic infections all around the world that may lead to hepatocellular carcinoma. This study is first time performed at the blood transfussion centre of Omar hospital, Lahore. It aims to determine the sero-prevalence of these diseases by screening the apparently healthy blood donors who might be the carriers of HBV or HCV and pose a high risk in the transmission. It also aims the comparison between the sensitivity of two diagnostic tests; chromatographic immunoassay – one step test device and Enzyme Linked Immuno Sorbant Assay (ELISA). Blood serum of 855 apparently healthy blood donors was screened for Hepatitis B surface antigen (HBsAg) and for anti HCV antibodies. SPSS version 12.0 and X2 (Chi-square) test were used for statistical analysis. The seroprevalence of HCV was 8.07% by the device method and by ELISA 9.12% and that of HBV was 5.6% by the device and 6.43% by ELISA. The unavailability of vaccination against HCV makes it more prevalent. Comparing the two diagnostic methods, ELISA proved to be more sensitive.

Keywords: ELISA, Sensitivity comparison of diagnostic tests, seroprevalence of Hepatitis B and C

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Authors: Gry M, Bergström J, Lengquist J, Lindberg J, Drobin K, Schwenk J, Nilsson P, Schuppe-Koistinen I.

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Sensitive and predictive DILI (Drug Induced Liver Injury) biomarkers are needed in drug R&D to improve early detection of hepatotoxicity. The discovery of DILI biomarkers that demonstrate the predictive power to identify individuals at risk to DILI would represent a major advance in the development of personalized healthcare approaches. In this healthy volunteer acetaminophen study (4g/day for 7 days, with 3 monitored nontreatment days before and 4 after), 450 serum samples from 32 subjects were analyzed using protein profiling by antibody suspension bead arrays. Multiparallel protein profiles were generated using a DILI target protein array with 300 antibodies, where the antibodies were selected based on previous literature findings of putative DILI biomarkers and a screening process using pre dose samples from the same cohort. Of the 32 subjects, 16 were found to develop an elevated ALT value (2Xbaseline, responders). Using the plasma profiling approach together with multivariate statistical analysis some novel findings linked to lipid metabolism were found and more important, endogenous protein profiles in baseline samples (prior to treatment) with predictive power for ALT elevations were identified.

Keywords: DILI, Plasma profiling, PLSDA, Randomforest.

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Authors: Robles F, Uribe A, Guadarrama A , Castellanos L, González C.

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The main objective of this study was to demonstrate that differentiation of infected and vaccinated animals (DIVA) strategy using different ELISA tests is possible when a subunit vaccine (Haemagglutinin protein) is used to prevent Avian influenza. Special emphasis was placed on the differentiation in the serological response to different components of the AIV (Nucleoprotein, Neuraminidase, Haemagglutinin, Nucleocapsid) between chickens that were vaccinated with a whole virus kill vaccine and recombinant vaccine. Furthermore, the potential use of this DIVA strategy using ELISA assays to detect Neuraminidase 1 (N1) was analyzed as strategy in countries where the field virus is H5N1 and the vaccine used is formulated with H5N2. Detection of AIV-s antibodies to any component in serum was negative for all animals on the study days 0-13. At study day 14 the titers of antibodies against Nucleoprotein (NP) and Nucleocapsid (NC) rose in the experimental groups vaccinated with Volvac® AI KV and were negatives during all the trial in the experimental groups vaccinated with a subunit H5; significant statistically differences were observed between these groups (p < 0.05). The seroconversion either Haemagglutinin or Neuraminidase was evident after 21 days post-vaccination in the experimental groups vaccinated with the respective viral fraction. Regarding the main aim of this study and according with the results that were obtained, use a combination of different ELISA test as a DIVA strategy is feasible when the vaccination is carry out with a subunit H5 vaccine. Also is possible to use the ELISA kit to detect Neuraminidase (either N1 or N2) as a DIVA concept in countries where H5N1 is present and the vaccination programs are done with H5N2 vaccine.

Keywords: Avian Influenza Virus, "DIVA concept", ELISAassay, subunit H5 vaccine.

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Authors: Mohammadjavad Mehrabanpour, Maryam Ranjbar Bushehri, Dorsa Mehrabanpour

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Respiratory diseases are the most important problems in the country’s poultry industry, particularly when it comes to broiler flocks. Ornithobacterium rhinotracheale (ORT) is a species that causes poor performance in growth rate, egg production, and mortality. This pathogen causes a respiratory infection including pulmonary alveolar inflammation, and pneumonia of birds throughout the world. Newcastle disease (ND) is a highly contagious disease in poultry, and also, it causes considerable losses to the poultry industry. The aim of this study was to evaluate the simultaneous occurrence of ORT and ND and NDV isolation by inoculation in embryonated eggs and confirmed by RT-PCR in broiler chicken flocks in Fars province. In this study, 318 blood and 85 tissue samples (brain, trachea, liver, and cecal tonsils) were collected from 15 broiler chicken farms. Survey serum antibody titers against ORT by using a commercial enzyme-linked immunosorbent assay (ELISA) kit performed. Evaluation of antibody titer against ND virus is performed by hemagglutination inhibition test. Virus isolation with chick embryo eggs 9-11 and RT-PCR method were carried out. A total of 318 serum samples, 135 samples (42.5%) were positive for antibodies to ORT and titer of HI antibodies against NDV in 122 serum samples (38/4%) were 7-10 (log2) and 61 serum samples (19/2%) had occurrence antibody titer against Newcastle virus and ORT. Results of the present study indicated that 20 tissue samples were positive in embryonated egg and in rapid hemagglutination (HA) test. HI test with specific ND positive serum confirmed that 6 of 20 samples. PCR confirmed that all six samples were positive and PCR products of samples indicated 535-base pair fragments in electrophrosis. Due to the great economic importance of these two diseases in the poultry industry, it is necessary to design and implement a comprehensive plan for prevention and control of these diseases.

Keywords: ELISA, Newcastle disease, Ornithobacterium rhinotracheale, seroprevalence.

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Authors: Y. Saylan, F. Yılmaz, A. Denizli

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In this study, we have focused our attention on combining of molecular imprinting into nanofilms and QCM nanosensor approaches and producing QCM nanosensor for anti- CCP, chosen as model protein, using anti-CCP imprinted nanofilms. The nonimprinted nanosensor was also prepared to evaluate the selectivity of the imprinted nanosensor. Anti-CCP imprinted QCM nanosensor was tested for real time detection of anti-CCP from aqueous solution. The kinetic and affinity studies were determined by using anti-CCP solutions with different concentrations. The responses related with mass shifts (%m) and frequency shifts (%f) were used to evaluate adsorption properties. To show the selectivity of the anti-CCP imprinted QCM nanosensor, competitive adsorption of anti-CCP and IgM was investigated. The results indicate that anti- CCP imprinted QCM nanosensor has higher adsorption capabilities for anti-CCP than for IgM, due to selective cavities in the polymer structure.

Keywords: Anti-CCP, molecular imprinting, QCM nanosensor, rheumatoid arthritis.

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Authors: Homa Torabizadeh

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This study proposes a basic molecular formula for all proteins. A total of 10,739 proteins belonging to 9 different protein groups classified on the basis of their functions were selected randomly. They included enzymes, storage proteins, hormones, signalling proteins, structural proteins, transport proteins, immunoglobulins or antibodies, motor proteins and receptor proteins. After obtaining the protein molecular formula using the ProtParam tool, the H/C, N/C, O/C, and S/C ratios were determined for each randomly selected sample. In this case, H, N, O, and S coefficients were specified per carbon atom. Surprisingly, the results demonstrated that H, N, O, and S coefficients for all 10,739 proteins are similar and highly correlated. This study demonstrates that despite differences in the structure and function, all known proteins have a similar basic molecular formula CnH1.58 ± 0.015nN0.28 ± 0.005nO0.30 ± 0.007nS0.01 ± 0.002n. The total correlation between all coefficients was found to be 0.9999.

Keywords: Protein molecular formula, Basic unit formula, Protparam tool.

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Authors: Li Li, Li-Ru Xia, He-Ye Wang, Xiao-Dong Bi

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In this paper, we presented a highly sensitive immune-affinity monolithic array for detection of α-fetoprotein (AFP) and carcinoembryonic antigen (CEA). Firstly, the epoxy functionalized monolith arrays were fabricated using UV initiated copolymerization method. Scanning electron microscopy (SEM) image showed that the poly(BABEA-co-GMA) monolith exhibited a well-controlled skeletal and well-distributed porous structure. Then, AFP and CEA immune-affinity monolithic arrays were prepared by immobilization of AFP and CEA antibodies on epoxy functionalized monolith arrays. With a non-competitive immune response format, the presented AFP and CEA immune-affinity arrays were demonstrated as an inexpensive, flexible, homogeneous and stable array for detection of AFP and CEA.

Keywords: Chemiluminescent detection, immune-affinity, monolithic copolymer array, UV-initiated copolymerization.

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Authors: Jeevan Malayan, Aparna Warrier, Padmasani Venkat Ramanan, Sanjeeva Reddy N, Elanchezhiyan Manickan

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MMR vaccine failure had been reported globally and here we report that it occurs now in India. Samples were collected from clinically suspected mumps cases were subjected for anti mumps antibodies, virus isolation, RT-PCR, sequencing and phylogenetic tree analysis. 56 samples collected from men and women belonging to various age groups. 30 had been vaccinated and the status of 26 patients was unknown. 28 out of 30 samples were found to be symptomatic and positive for Mumps IgM, indicating active mumps infection in 93.4% of the vaccinated population. A phylogenetic tree comparison of the clinical isolate is shown to be genotype C which is distinct from vaccine strain. Our study clearly sending warning signs that MMR vaccine is a failure and it needs to be revamped for the human use by increasing its efficacy and efficiency.

Keywords: Genotype C, Mumps virus, MMR vaccine, Sero types.

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Authors: Tomohiro Hachino, Kengo Nagatomo, Hitoshi Takata

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This paper deals with an on-line identification method of continuous-time Hammerstein systems by using the radial basis function (RBF) networks and immune algorithm (IA). An unknown nonlinear static part to be estimated is approximately represented by the RBF network. The IA is efficiently combined with the recursive least-squares (RLS) method. The objective function for the identification is regarded as the antigen. The candidates of the RBF parameters such as the centers and widths are coded into binary bit strings as the antibodies and searched by the IA. On the other hand, the candidates of both the weighting parameters of the RBF network and the system parameters of the linear dynamic part are updated by the RLS method. Simulation results are shown to illustrate the proposed method.

Keywords: Continuous-time System, Hammerstein System, OnlineIdentification, Immune Algorithm, RBF network.

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Authors: Marwa I. Shabayek, Ola A. Said, Hanan A. Attaia, Heba A. Awida

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Bladder carcinoma is an important worldwide health problem. Both cystoscopy and urine cytology used in detecting bladder cancer suffer from drawbacks where cystoscopy is an invasive method and urine cytology shows low sensitivity in low grade tumors. This study validates easier and less time-consuming techniques to evaluate the value of combined use of angiogenin and clusterin in comparison and combination with voided urine cytology in the detection of bladder cancer patients. This study includes malignant (bladder cancer patients, n= 50), benign (n=20) and healthy (n=20) groups. The studied groups were subjected to cystoscopic examination, detection of bilharzial antibodies, urine cytology, and estimation of urinary angiogenin and clusterin by ELISA. The overall sensitivity and specificity were 66% and 75% for angiogenin, 70% and 82.5% for clusterin and 46% and 80% for voided urine cytology. Combined sensitivity of angiogenin and clusterin with urine cytology increased from 82 to 88%. 

Keywords: Angiogenin, Bladder Cancer, Clusterin, Cytology.

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Authors: Ramin Javadzadeh., Zahra Afsahi, MohammadReza Meybodi

Abstract:

The Artificial immune systems algorithms are Meta heuristic optimization method, which are used for clustering and pattern recognition applications are abundantly. These algorithms in multimodal optimization problems are more efficient than genetic algorithms. A major drawback in these algorithms is their slow convergence to global optimum and their weak stability can be considered in various running of these algorithms. In this paper, improved Artificial Immune System Algorithm is introduced for the first time to overcome its problems of artificial immune system. That use of the small size of a local search around the memory antibodies is used for improving the algorithm efficiently. The credibility of the proposed approach is evaluated by simulations, and it is shown that the proposed approach achieves better results can be achieved compared to the standard artificial immune system algorithms

Keywords: Artificial immune system, Cellular Automata, Cellular learning automata, Cellular learning automata, , Local search, Optimization.

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Authors: H.K. Abd El-Maksoud, H.H. Keyser

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Competitive relationships among Bradyrhizobium japonicum USDA serogroup 123, 122 and 138 were screened versus the standard commercial soybean variety Williams and two introductions P1 377578 "671" in a field trial. Displacement of strain 123 by an effective strain should improved N2 fixation. Root nodules were collected and strain occupancy percentage was determined using strain specific fluorescent antibodies technique. As anticipated the strain USDA 123 dominated 92% of nodules due to the high affinity between the host and the symbiont. This dominance was consistent and not changed materially either by inoculation practice or by introducing new strainan. The interrelationship between the genotype Williams and serogroup 122 & 138 was found very weak although the cell density of the strain in the rhizosphere area was equal. On the other hand, the nodule occupancy of genotypes 671 and 166 with rhizobia serogroup 123 was almost diminished to zero. . The data further exhibited that the genotypes P1 671 and P1 166 have high affinity to colonize with strains 122 and 138 whereas Williams was highly promiscuous to strain 123.

Keywords: B. japonicum serogroups, Competition, Host restriction, Soybean genotype.

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Authors: Wanli Ma, Dat Tran, Dharmendra Sharma

Abstract:

The temporal nature of negative selection is an under exploited area. In a negative selection system, newly generated antibodies go through a maturing phase, and the survivors of the phase then wait to be activated by the incoming antigens after certain number of matches. These without having enough matches will age and die, while these with enough matches (i.e., being activated) will become active detectors. A currently active detector may also age and die if it cannot find any match in a pre-defined (lengthy) period of time. Therefore, what matters in a negative selection system is the dynamics of the involved parties in the current time window, not the whole time duration, which may be up to eternity. This property has the potential to define the uniqueness of negative selection in comparison with the other approaches. On the other hand, a negative selection system is only trained with “normal" data samples. It has to learn and discover unknown “abnormal" data patterns on the fly by itself. Consequently, it is more appreciate to utilize negation selection as a system for pattern discovery and recognition rather than just pattern recognition. In this paper, we study the potential of using negative selection in discovering unknown temporal patterns.

Keywords: Artificial Immune Systems, ComputationalIntelligence, Negative Selection, Pattern Discovery.

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Authors: M. Hashemi, R. Madani, N. Razmi

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M. paratuberculosis is a slow growing mycobactin dependent mycobacterial species known to be the causative agent of Johne’s disease in all species of domestic ruminants worldwide. JD is characterized by gradual weight loss; decreased milk production. Excretion of the organism may occur for prolonged periods (1 to 2.5 years) before the onset of clinical disease. In recent years researchers focus on identification a specific antigen of MAP to use in diagnosis test and preparation of effective vaccine. In this paper, for production of polyclonal antibody against proteins of Mycobacterium avium paratuberculosis cell well a rabbit immunization at a certain time period with antigen. After immunization of the animal, rabbit was bleeded for producing enriched serum. Antibodies were purification with ion exchange chromatography. For exact measurement of interaction, western blotting test was used that this study demonstrated sharp bands appears in nitrocellulose paper and specific bands were 50 and 150 KD molecular weight. These were indicating immunodominant proteins.

Keywords: Paratuberculosis, Immunodominant, Western blotting, Ion exchange choromatography.

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Authors: A. Asambe

Abstract:

A study was conducted to determine the level of compliance with sanitary measures in piggeries, and awareness and risk factors of African swine fever in Benue State, Nigeria. Questionnaires were distributed to 74 respondents consisting of piggery owners and attendants in different piggeries across 12 LGAs to collect data for this study. Sanitary measures in piggeries were observed to be generally very poor, though respondents admitted being aware of ASF. Piggeries located within a 1 km radius of a slaughter slab (OR=9.2, 95% CI - 3.0-28.8), piggeries near refuse dump sites (OR=3.0, 95% CI - 1.0-9.5) and piggeries where farm workers wear their work clothes outside of the piggery premises (OR=0.2, 95% CI - 0.1-0.7) showed higher chances of ASFV infection and were significantly associated (p < 0.0001), (p < 0.05) and (p < 0.01), and were identified as potential risk factors. The study concluded that pigs in Benue State are still at risk of an ASF outbreak. Proper sanitary and hygienic practices is advocated and emphasized in piggeries, while routine surveillance for ASFV antibodies in pigs in Benue State is strongly recommended to provide a reliable reference data base to plan for the prevention of any devastating ASF outbreak.

Keywords: African swine fever, awareness, piggery, risk factors, sanitary measures.

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Authors: A.S. Sikarwar, S. Ambu, T .H. Wong

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Snake bite cases in Malaysia most often involve the species Naja-naja and Calloselasma rhodostoma. In keeping with the need for a rapid snake venom detection kit in a clinical setting, plate and dot-ELISA test for the venoms of Naja-naja sumatrana, Calloselasma rhodostoma and the cobra venom fraction V antigen was developed. Polyclonal antibodies were raised and further used to prepare the reagents for the dot-ELISA test kit which was tested in mice, rabbit and virtual human models. The newly developed dot- ELISA kit was able to detect a minimum venom concentration of 244ng/ml with cross reactivity of one antibody type. The dot-ELISA system was sensitive and specific for all three snake venom types in all tested animal models. The lowest minimum venom concentration detectable was in the rabbit model, 244ng/ml of the cobra venom fraction V antigen. The highest minimum venom concentration was in mice, 1953ng/ml against a multitude of venoms. The developed dot-ELISA system for the detection of three snake venom types was successful with a sensitivity of 95.8% and specificity of 97.9%.

Keywords: ELISA, Venom, SVDK, Naja-naja sumatrana , Calloselasma rhodostoma.

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Authors: Stela Papa, Klementina Puto, Migena Pllaha

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HCV is a hepatotropic RNA virus, transmitted primarily via the blood route, which causes progressive disease such as chronic hepatitis, liver cirrhosis, or hepatocellular carcinoma. HCV nowadays is a global healthcare problem. A variety of immunoassays including old and new technologies are being applied to detect HCV in our country. These methods include Immunochromatography assays (ICA), Fluorescence immunoassay (FIA), Enzyme linked fluorescent assay (ELFA), and Enzyme linked immunosorbent assay (ELISA) to detect HCV antibodies in blood serum, which lately is being slowly replaced by more sensitive methods such as rapid automated analyzer chemiluminescence immunoassay (CLIA). The aim of this study is to estimate HCV infection in carriers and chronic acute patients and to evaluate the use of new diagnostic methods. This study was realized from September 2016 to May 2018. During this study period, 2913 patients were analyzed for the presence of HCV by taking samples from their blood serum. The immunoassays performed were ICA, FIA, ELFA, ELISA, and CLIA assays. Concluding, 82% of patients taken in this study, resulted infected with HCV. Diagnostic methods in clinical laboratories are crucial in the early stages of infection, in the management of chronic hepatitis and in the treatment of patients during their disease.

Keywords: CLIA, ELISA, hepatitis C virus, immunoassay.

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Authors: F. Kashanian, M. M. Masoudi, A. Akbari, A. Shamloo, M. R. Zand, S. S. Salehi

Abstract:

Nano-sized materials present new opportunities in biology and medicine and they are used as biomedical tools for investigation, separation of molecules and cells. To achieve more effective cancer therapy, it is essential to select cancer cells exactly. This research suggests that using the antibody-functionalized nontoxic Arginine-doped magnetic nanoparticles (A-MNPs), has been prosperous in detection, capture, and magnetic separation of circulating tumor cells (CTCs) in tumor tissue. In this study, A-MNPs were synthesized via a simple precipitation reaction and directly immobilized Ep-CAM EBA-1 antibodies over superparamagnetic A-MNPs for Mucin BCA-225 in breast cancer cell. The samples were characterized by vibrating sample magnetometer (VSM), FT-IR spectroscopy, Tunneling Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM). These antibody-functionalized nontoxic A-MNPs were used to capture breast cancer cell. Through employing a strong permanent magnet, the magnetic separation was achieved within a few seconds. Antibody-Conjugated nontoxic Arginine-doped Fe₃O₄ nanoparticles have the potential for the future study to capture CTCs which are released from tumor tissue and for drug delivery, and these results demonstrate that the antibody-conjugated A-MNPs can be used in magnetic hyperthermia techniques for cancer treatment.

Keywords: Tumor tissue, antibody, magnetic nanoparticle, CTCs capturing.

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Authors: Alaa A. Shamaun Al-Abboodi, Yunis A. A. Bapeer

Abstract:

400 one-day-old male broiler chicks (Ross-308) randomly divided to 2 main groups, 1st main group (GA) was feeding basal diet with medium chain fatty acid (MCFA) at rate of 0.15% and divided to four subgroups, 3 subgroups vaccinated with different routes with Newcastle Disease Virus (NDV) and non-vaccinated group. The 2nd main group (GB) feeding basal diet without MCFA and divided the same as 1st main group. The parameters used in this study included: ND antibody titers at 1, 10, 21, 28, 35 and 42 days of age and values of CD4 and CD8 at 1, 20, 30 and 42 days of age. This experiment detected increase in ND antibodies titers in (G1, G2, G3) groups were fed on basal diet MCFA comparing to groups were fed without adding MCFA (G5, G6, G7) and control groups (G4, G8). The results of cellular immune response (CD4 and CD8) T-cells in broiler chicks indicated that there was obviously significant relationship between dietary Fatty Acid (FA) versus the diet without FA on the level of CD4 parameter, for the entire experimental period. The effect of different ages was statistically significant in creating different values of CD4 level, whereas the CD4 level decreases markedly with age. However, analyzing the data of different vaccination methods, oculonasal method of vaccination led to the highest value of CD4 compared with the oral, S/C and control groups. There were statistical differences in CD8 values due to supplementation of FA versus the basal diet and due to the effect of different age periods. As for the age effect, the CD8 value at 20 days of age was significantly higher than at 42 and 30 days.

Keywords: Broiler, CD4 and CD8, fatty acids, Newcastle disease.

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Authors: Srinivas Bathini, Duraichelvan Raju, Simona Badilescu, Muthukumaran Packirisamy

Abstract:

A bio-sensing method, based on the plasmonic property of gold nano-islands, has been developed for detection of exosomes in a clinical setting. The position of the gold plasmon band in the UV-Visible spectrum depends on the size and shape of gold nanoparticles as well as on the surrounding environment. By adsorbing various chemical entities, or binding them, the gold plasmon band will shift toward longer wavelengths and the shift is proportional to the concentration. Exosomes transport cargoes of molecules and genetic materials to proximal and distal cells. Presently, the standard method for their isolation and quantification from body fluids is by ultracentrifugation, not a practical method to be implemented in a clinical setting. Thus, a versatile and cutting-edge platform is required to selectively detect and isolate exosomes for further analysis at clinical level. The new sensing protocol, instead of antibodies, makes use of a specially synthesized polypeptide (Vn96), to capture and quantify the exosomes from different media, by binding the heat shock proteins from exosomes. The protocol has been established and optimized by using a glass substrate, in order to facilitate the next stage, namely the transfer of the protocol to a microfluidic environment. After each step of the protocol, the UV-Vis spectrum was recorded and the position of gold Localized Surface Plasmon Resonance (LSPR) band was measured. The sensing process was modelled, taking into account the characteristics of the nano-island structure, prepared by thermal convection and annealing. The optimal molar ratios of the most important chemical entities, involved in the detection of exosomes were calculated as well. Indeed, it was found that the results of the sensing process depend on the two major steps: the molar ratios of streptavidin to biotin-PEG-Vn96 and, the final step, the capture of exosomes by the biotin-PEG-Vn96 complex. The microfluidic device designed for sensing of exosomes consists of a glass substrate, sealed by a PDMS layer that contains the channel and a collecting chamber. In the device, the solutions of linker, cross-linker, etc., are pumped over the gold nano-islands and an Ocean Optics spectrometer is used to measure the position of the Au plasmon band at each step of the sensing. The experiments have shown that the shift of the Au LSPR band is proportional to the concentration of exosomes and, thereby, exosomes can be accurately quantified. An important advantage of the method is the ability to discriminate between exosomes having different origins.

Keywords: Exosomes, gold nano-islands, microfluidics, plasmonic biosensing.

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