Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 35

Search results for: antibody

35 Comparison of Anti-Shadoo Antibodies – Where is the Endogenous Shadoo protein?

Authors: Eszter Tóth, Ervin Welker

Abstract:

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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34 Dengue Transmission Model between Infantand Pregnant Woman with Antibody

Authors: R. Kongnuy, P. Pongsumpun

Abstract:

Dengue, a disease found in most tropical and subtropical areas of the world. It has become the most common arboviral disease of humans. This disease is caused by any of four serotypes of dengue virus (DEN1-DEN4). In many endemic countries, the average age of getting dengue infection is shifting upwards, dengue in pregnancy and infancy are likely to be encountered more frequently. The dynamics of the disease is studied by a compartmental model involving ordinary differential equations for the pregnant, infant human and the vector populations. The stability of each equilibrium point is given. The epidemic dynamic is discussed. Moreover, the numerical results are shown for difference values of dengue antibody.

Keywords: Dengue antibody, infant, pregnant human, mathematical model.

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33 Dextran Modified Silicon Photonic Microring Resonator Sensors

Authors: Jessie Yiying Quah, Vivian Netto, Jack Sheng Kee, Eric Mouchel La Fosse, Mi Kyoung Park

Abstract:

We present a dextran modified silicon microring resonator sensor for high density antibody immobilization. An array of sensors consisting of three sensor rings and a reference ring was fabricated and its surface sensitivity and the limit of detection were obtained using polyelectrolyte multilayers. The mass sensitivity and the limit of detection of the fabricated sensor ring are 0.35 nm/ng mm-2 and 42.8 pg/mm2 in air, respectively. Dextran modified sensor surface was successfully prepared by covalent grafting of oxidized dextran on 3-aminopropyltriethoxysilane (APTES) modified silicon sensor surface. The antibody immobilization on hydrogel dextran matrix improves 40% compared to traditional antibody immobilization method via APTES and glutaraldehyde linkage.

Keywords: Antibody immobilization, Dextran, Immunosensor, Label-free detection, Silicon micro-ring resonator

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32 Antibody-Conjugated Nontoxic Arginine-Doped Fe3O4 Nanoparticles for Magnetic Circulating Tumor Cells Separation

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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31 ELISA Based hTSH Assessment Using Two Sensitive and Specific Anti-hTSH Polyclonal Antibodies

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

Abstract:

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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30 An Immunosensor for Bladder Cancer Screening

Authors: Congo Tak Shing Ching, Hong-Sheng Chen, Tai-Ping Sun, Hsiu-Li Shieh

Abstract:

Nuclear matrix protein 22 (NMP22) is a FDA approved biomarker for bladder cancer. The objective of this study is to develop a simple NMP22 immumosensor (NMP22-IMS) for accurate measurement of NMP22. The NMP22-IMS was constructed with NMP22 antibody immobilized on screen-printed carbon electrodes. The construction procedures and antibody immobilization are simple. Results showed that the NMP22-IMS has an excellent (r2³0.95) response range (20 – 100 ng/mL). In conclusion, a simple and reliable NMP22-IMS was developed, capable of precisely determining urine NMP22 level.

Keywords: Bladder Cancer, Immunosensor, Impedance, NMP22

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29 Antibody Reactivity of Synthetic Peptides Belonging to Proteins Encoded by Genes Located in Mycobacterium tuberculosis-Specific Genomic Regions of Differences

Authors: Abu Salim Mustafa

Abstract:

The comparisons of mycobacterial genomes have identified several Mycobacterium tuberculosis-specific genomic regions that are absent in other mycobacteria and are known as regions of differences. Due to M. tuberculosis-specificity, the peptides encoded by these regions could be useful in the specific diagnosis of tuberculosis. To explore this possibility, overlapping synthetic peptides corresponding to 39 proteins predicted to be encoded by genes present in regions of differences were tested for antibody-reactivity with sera from tuberculosis patients and healthy subjects. The results identified four immunodominant peptides corresponding to four different proteins, with three of the peptides showing significantly stronger antibody reactivity and rate of positivity with sera from tuberculosis patients than healthy subjects. The fourth peptide was recognized equally well by the sera of tuberculosis patients as well as healthy subjects. Predication of antibody epitopes by bioinformatics analyses using ABCpred server predicted multiple linear epitopes in each peptide. Furthermore, peptide sequence analysis for sequence identity using BLAST suggested M. tuberculosis-specificity for the three peptides that had preferential reactivity with sera from tuberculosis patients, but the peptide with equal reactivity with sera of TB patients and healthy subjects showed significant identity with sequences present in nob-tuberculous mycobacteria. The three identified M. tuberculosis-specific immunodominant peptides may be useful in the serological diagnosis of tuberculosis.

Keywords: Genomic regions of differences, Mycobacterium tuberculosis, peptides, serodiagnosis.

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28 Displaying of GnRH Peptides on Bacteriophage T7 and Its Immunogenicity in Mice Model

Authors: Hai Xu, Yiwei Wang, Xi Bao, Bihua Deng, Pengcheng Li, Yu Lu

Abstract:

T7 phage could be used as a perfect vector for peptides expression and haptens presentation. T7-3GnRH recombinant phage was constructed by inserting three copies of Gonadotrophin Releasing Hormone (GnRH) gene into the multiple cloning site of T7 Select 415-1b phage genome. The positive T7-3GnRH phage was selected by using polymerase chain reaction amplification, and the p10B-3GnRH fusion protein was verified by SDS-PAGE and Western-blotting assay. T7-3GnRH vaccine was made and immunized with 1010 pfu in 0.2 ml per dose in mice. Blood samples were collected at an interval in weeks, and anti-GnRH antibody and testosterone concentrations were detected by ELISA and radioimmunoassay, respectively. The results show that T7-3GnRH phage particles confer a high immunogenicity to the GnRH-derived epitope. Moreover, the T7-3GnRH vaccine induced higher level of anti-GnRH antibody than ImproVac®. However, the testosterone concentrations in both immunized groups were at a similar level, and the testis developments were significantly inhibited compared to controls. These findings demonstrated that the anti-GnRH antibody could neutralize the endogenous GnRH to down regulate testosterone level and limit testis development, highlighting the potential value of T7-3GnRH in the immunocastration vaccine research.

Keywords: Gonadotrophin releasing hormone, GnRH, immunocastration, T7 phage, phage vaccine.

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27 Quality Based Approach for Efficient Biologics Manufacturing

Authors: Takashi Kaminagayoshi, Shigeyuki Haruyama

Abstract:

To improve the manufacturing efficiency of biologics, such as antibody drugs, a quality engineering framework was designed. Within this framework, critical steps and parameters in the manufacturing process were studied. Identification of these critical steps and critical parameters allows a deeper understanding of manufacturing capabilities, and suggests to process development department process control standards based on actual manufacturing capabilities as part of a PDCA (plan-do-check-act) cycle. This cycle can be applied to each manufacturing process so that it can be standardized, reducing the time needed to establish each new process.

Keywords: Antibody drugs, biologics, manufacturing efficiency, PDCA cycle, quality engineering.

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26 Development of an Immunoassay Platform for Diagnosis of Acute Kidney Injury

Authors: T. Bovornvirakit, K. Viravaidya

Abstract:

Acute kidney injury (AKI) is a new worldwide public health problem. A diagnosis of this disease using creatinine is still a problem in clinical practice. Therefore, a measurement of biomarkers responsible for AKI has received much attention in the past couple years. Cytokine interleukin-18 (IL-18) was reported as one of the early biomarkers for AKI. The most commonly used method to detect this biomarker is an immunoassay. This study used a planar platform to perform an immunoassay using fluorescence for detection. In this study, anti-IL-18 antibody was immobilized onto a microscope slide using a covalent binding method. Make-up samples were diluted at the concentration between 10 to 1000 pg/ml to create a calibration curve. The precision of the system was determined using a coefficient of variability (CV), which was found to be less than 10%. The performance of this immunoassay system was compared with the measurement from ELISA.

Keywords: Acute kidney injury, Acute renal failure, Antibody immobilization, Interleukin-18

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25 Hybrid Artificial Immune System for Job Shop Scheduling Problem

Authors: Bin Cai, Shilong Wang, Haibo Hu

Abstract:

The job shop scheduling problem (JSSP) is a notoriously difficult problem in combinatorial optimization. This paper presents a hybrid artificial immune system for the JSSP with the objective of minimizing makespan. The proposed approach combines the artificial immune system, which has a powerful global exploration capability, with the local search method, which can exploit the optimal antibody. The antibody coding scheme is based on the operation based representation. The decoding procedure limits the search space to the set of full active schedules. In each generation, a local search heuristic based on the neighborhood structure proposed by Nowicki and Smutnicki is applied to improve the solutions. The approach is tested on 43 benchmark problems taken from the literature and compared with other approaches. The computation results validate the effectiveness of the proposed algorithm.

Keywords: Artificial immune system, Job shop scheduling problem, Local search, Metaheuristic algorithm

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24 The Role of Immunogenic Adhesin Vibrio alginolyticus 49 k Da to Molecule Expression of Major Histocompatibility Complex on Receptors of Humpback Grouper Cromileptes altivelis

Authors: Uun Yanuhar

Abstract:

The purpose of research was to know the role of immunogenic protein of 49 kDa from V.alginolyticus which capable to initiate molecule expression of MHC Class II in receptor of Cromileptes altivelis. The method used was in vivo experimental research through testing of immunogenic protein 49 kDa from V.alginolyticus at Cromileptes altivelis (size of 250 - 300 grams) using 3 times booster by injecting an immunogenic protein in a intramuscular manner. Response of expressed MHC molecule was shown using immunocytochemistry method and SEM. Results indicated that adhesin V.alginolyticus 49 kDa which have immunogenic character could trigger expression of MHC class II on receptor of grouper and has been proven by staining using immunocytochemistry and SEM with labeling using antibody anti MHC (anti mouse). This visible expression based on binding between epitopes antigen and antibody anti MHC in the receptor. Using immunocytochemistry, intracellular response of MHC to in vivo induction of immunogenic adhesin from V.alginolyticus was shown.

Keywords: C.altivelis, immunogenic, MHC, V.alginolyticus.

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23 Colorectal Cancer Screening by a CEACAM-6 Immunosensor

Authors: C. T. S. Ching, P. W. C hen, T. P. Sun, H. L. Shieh

Abstract:

The biomarker for colorectal cancer (CRC) is CEACAM-6 antigen (C6AG). Therefore, this study aims to develop a novel, simple and low-cost CEACAM-6 antigen immumosensor (C6AG-IMS), based on electrical impedance measurement, for precise determination of C6AG. A low-cost screen-printed graphite electrode was constructed and used as the sensor, with CEACAM-6 antibody (C6AB) immobilized on it. The procedures of sensor fabrication and antibody immobilization are simple and low-cost. Measurement of the electrical impedance at a definite frequency ranges (0.43 – 1.26 MHz) showed that the C6AG-IMS has an excellent linear (r2>0.9) response range (8.125 – 65 pg/mL), covering the normal physiological and pathological ranges of blood C6AG levels. Also, the C6AG-IMS has excellent reliability and validity, with the intraclass correlation coefficient being 0.97. In conclusion, a novel, simple, low-cost and reliable C6AG-IMS was designed and developed, being able to accurately determine blood C6AG levels in the range of pathological and normal physiological regions. The C6AG-IMS can provide a point-of-care and immediate screening results to the user at home.

Keywords: Colorectal Cancer, Immunosensor, Electrical Impedance, CEACAM-6, Measurement, Point-of-Care

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22 Molecular and Serological Diagnosis of Newcastle and Ornithobacterium rhinotracheale Broiler in Chicken in Fars Province, Iran

Authors: Mohammadjavad Mehrabanpour, Maryam Ranjbar Bushehri, Dorsa Mehrabanpour

Abstract:

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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21 Chitosan Nanoparticle as a Novel Delivery System for A/H1n1 Influenza Vaccine: Safe Property and Immunogenicity in Mice

Authors: Nguyen Anh Dzung, Nguyen Thi Ngoc Hà, Dang Thi Hong Van, Nguyen Thi Lan Phuong, Nguyen Thi Nhu Quynh, Dinh Minh Hiep, Le Van Hiep

Abstract:

The aims of this paper are to study the efficacy of chitosan nanoparticles in stimulating specific antibody against A/H1N1 influenza antigen in mice. Chitosan nanoparticles (CSN) were characterized by TEM. The results showed that the average size of CSN was from 80nm to 106nm. The efficacy of A/H1N1 influenza vaccine loaded on the surface of CSN showed that loading efficiency of A/H1N1 influenza antigen on CSN was from 93.75 to 100%. Safe property of the vaccine were tested. In 10 days post vaccination, group of CSN 30 kDa and 300 kDa loaded A/H1N1 influenza antigen were the rate of immune response on mice to be 100% (9/9) higher than Al(OH)3 and other adjuvant. 100% mice in the experiment of all groups had immune response in 20 days post vaccination. The results also showed that HI titer of the group using CSN 300 kDa as an adjuvant increased significantly up to 3971 HIU, over three-fold higher than the Al(OH)3 adjuvant, chitosan (CS), and one hundredfold than the A/H1N1 antigen only. Stability of the vaccine formulation was investigated.

Keywords: Chitosan nanoparticles, A/H1N1 influenza antigen, vaccine, immunogenicity, adjuvant, antibody titer

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20 The Effect of Different Levels of Seed and Extract of Harmal (Peganum harmala L.) on Immune Responses of Broiler Chicks

Authors: M. Toghyani, A. Ghasemi, S. A. Tabeidian

Abstract:

The present study was carried out to evaluate the effect of different levels of dietary seed and extract of Harmal (Peganum harmala L.) on immunity of broiler chicks. A total of 350 one-day old broiler chicks (Ross 308) were randomly allocated to five dietary treatments with four replicates pen of 14 birds each. Dietary treatments consisted of control, 1 and 2 g/kg Harmal seed in diet, 100 and 200 mg/L Harmal seed extract in water. Broilers received dietary treatments from 1 to 42 d. Two birds from each pen were randomly weighed and sacrificed at 42 d of age, the relative weight of lymphoid organs (bursa of Fabercius and spleen) to live weight were calculated. Antibody titers against Newcastle and influenza viruses and sheep red blood cell were measured at 30 d of age. Results showed that the relative weights of lymphoid organs were not affected by dietary treatments. Furthermore, antibody titer against Newcastle and influenza viruses as well as sheep red blood cell antigen were significantly (P<0.05) enhanced by feeding Harmal seed and extract. In conclusion, the results indicated that dietary inclusion of Harmal seed and extract enhanced immunological responses in broiler chicks.

Keywords: Broiler chicks, Harmal, immunity.

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19 Effect of Fatty Acids in Feed on Levels of Antibody Titers and CD4 and CD8 T-Lymphocyte against Newcastle Disease Virus of Vaccinated Broiler Chicken

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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18 Outcomes of Pregnancy in Women with TPO Positive Status after Appropriate Dose Adjustments of Thyroxin: A Prospective Cohort Study

Authors: Revathi S. Rajan, Pratibha Malik, Nupur Garg, Smitha Avula, Kamini A. Rao

Abstract:

This study aimed to analyse the pregnancy outcomes in patients with TPO positivity after appropriate L-Thyroxin supplementation with close surveillance. All pregnant women attending the antenatal clinic at Milann-The Fertility Center, Bangalore, India- from Aug 2013 to Oct 2014 whose booking TSH was more than 2.5 mIU/L were included along with those pregnant women with prior hypothyroidism who were TPO positive. Those with TPO positive status were vigorously managed with appropriate thyroxin supplementation and the doses were readjusted every 3 to 4 weeks until delivery. Women with recurrent pregnancy loss were also tested for TPO positivity and if tested positive, were monitored serially with TSH and fT4 levels every 3 to 4 weeks and appropriately supplemented with thyroxin when the levels fluctuated. The testing was done after an informed consent in all these women. The statistical software namely SAS 9.2, SPSS 15.0, Stata 10.1, MedCalc 9.0.1, Systat 12.0 and R environment ver.2.11.1 were used for the analysis of the data. 460 pregnant women were screened for thyroid dysfunction at booking of which 52% were hypothyroid. Majority of them (31.08%) were subclinically hypothyroid and the remaining were overt. 25% of the total no. of patients screened were TPO positive. The various pregnancy complications that were observed in the TPO positive women were gestational glucose intolerance [60%], threatened abortion [21%], midtrimester abortion [4.3%], premature rupture of membranes [4.3%], cervical funneling [4.3%] and fetal growth restriction [3.5%]. 95.6% of the patients who followed up till the end delivered beyond 30 weeks. 42.6% of these patients had previous history of recurrent abortions or adverse obstetric outcome and 21.7% of the delivered babies required NICU admission. Obstetric outcomes in our study in terms of midtrimester abortions, placental abruption, and preterm delivery improved for the better after close monitoring of the thyroid hormone [TSH and fT4] levels every 3 to 4 weeks with appropriate dose adjustment throughout pregnancy. Euthyroid women with TPO positive status enrolled in the study incidentally were those with recurrent abortions/infertility and required thyroxin supplements due to elevated Thyroid hormone (TSH, fT4) levels during the course of their pregnancy. Significant associations were found with age>30 years and Hyperhomocysteinemia [p=0.017], recurrent pregnancy loss or previous adverse obstetric outcomes [p=0.067] and APLA [p=0.029]. TPO antibody levels >600 I U/ml were significantly associated with development of gestational hypertension [p=0.041] and fetal growth restriction [p=0.082]. Euthyroid women with TPO positivity were also screened periodically to counter fluctuations of the thyroid hormone levels with appropriate thyroxin supplementation. Thus, early identification along with aggressive management of thyroid dysfunction and stratification of these patients based on their TPO status with appropriate thyroxin supplementation beginning in the first trimester will aid risk modulation and also help avert complications.

Keywords: Antinuclear antibody, Subclinical hypothyroidism, Thyroxin, TPO antibody.

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17 Effect of Dietary Supplementation of Different Levels of Black Seed (Nigella Sativa L.) on Growth Performance, Immunological, Hematological and Carcass Parameters of Broiler Chicks

Authors: R. S. Shewita, A. E. Taha

Abstract:

This experiment was conducted to investigate the effect of dietary supplementation of different levels of black seed (Nigella sativa L.) on the performance and immune response of broiler chicks. A total 240 day-old broiler chicks were used and randomly allotted equally into six experimental groups designated as 1, 2, 3, 4, 5 and 6 having black seed at the rate of 0, 2, 4, 6, 8 and 10 g /kg diet respectively. The study was lasted for 42 days. Average body weight, weight gain, relative growth rate, feed conversion, antibody titer against Newcastle disease, phagocytic activity and phagocytic index, some blood parameters(GOT, GPT, Glucose, Cholesterol, Triglyceride, Total protein, Albumen, WBCs, RBCs, Hb and PCV), dressing percentage, weight of different body organs, abdominal fat weight, were determined. It was found that, N. Sativa significantly improved final body weight, total body gain and feed conversion ratio of groups 2 and 3 when compared with the control group. Higher levels of N. Sativa did not improve growth performance of the chicks. Non significant differences were observed for antibody titer against Newcastle virus, WBCs count, serum GOT, glucose level, dressing %, relative liver, spleen, heart and head percentages. Lymphoid organs (Bursa and Thymus) improved significantly with increasing N. Sativa level in all supplemented groups. Serum cholesterol, triglyceride and visible fat % significantly decreased with Nigella sativa supplementation while serum GPT level significantly increased with nigella sativa supplementation.

Keywords: Nigella Sativa, broiler, growth, carcass traits, serum, blood

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16 Controlled Assembly of Magnetic Biomolecular Nanostructures

Authors: Hui Wang, Harish Padmanabhan, David Thomson, Krassen Dimitrov

Abstract:

Two optimized strategies were successfully established to develop biomolecule-based magnetic nanoassemblies. Streptavidin-coated and amine-coated magnetic nanoparticles were chosen as model scaffolds onto which double-stranded DNA and human immunoglobulin G were specifically conjugated in succession, using biotin-streptavidin interaction or covalent cross-linkers. The success of this study opens the prospect of developing selective and sensitive nanoparticle-based structures for diagnostics or drug delivery.

Keywords: Antibody, DNA, linker, magnetic nanoparticles, streptavidin

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15 Quantitative Analysis of Carcinoembryonic Antigen (CEA) Using Micromechanical Piezoresistive Cantilever

Authors: Meisam Omidi, M. Mirijalili, Mohammadmehdi Choolaei, Z. Sharifi, F. Haghiralsadat, F. Yazdian

Abstract:

In this work, we have used arrays of micromechanical piezoresistive cantilever with different geometries to detect carcinoembryonic antigen (CEA), which is known as an important biomarker associated with various cancers such as colorectal, lung, breast, pancreatic, and bladder cancer. The sensing principle is based on the surface stress changes induced by antigen–antibody interaction on the microcantilevers surfaces. Different concentrations of CEA in a human serum albumin (HSA) solution were detected as a function of deflection of the beams. According to the experiments, it was revealed that microcantilevers have surface stress sensitivities in the order of 8 (mJ/m). This matter allows them to detect CEA concentrations as low as 3 ng/mL or 18 pM. This indicates the fact that the self-sensing microcantilevers approach is beneficial for pathological tests.

Keywords: Micromechanical biosensors, Carcinoembryonic antigen (CEA), surface stress.

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14 Rapid Detection System of Airborne Pathogens

Authors: Shigenori Togashi, Kei Takenaka

Abstract:

We developed new processes which can collect and detect rapidly airborne pathogens such as the avian flu virus for the pandemic prevention. The fluorescence antibody technique is known as one of high-sensitive detection methods for viruses, but this needs up to a few hours to bind sufficient fluorescence dyes to viruses for detection. In this paper, we developed a mist-labeling can detect substitution viruses in a short time to improve the binding rate of fluorescent dyes and substitution viruses by the micro reaction process. Moreover, we developed the rapid detection system with the above “mist labeling”. The detection system set with a sampling bag collecting patient’s breath and a cartridge can detect automatically pathogens within 10 minutes.

Keywords: Viruses, Sampler, Mist, Detection, Fluorescent dyes, Microreaction.

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13 Combine a Population-based Incremental Learning with Artificial Immune System for Intrusion Detection System

Authors: Jheng-Long Wu, Pei-Chann Chang, Hsuan-Ming Chen

Abstract:

This research focus on the intrusion detection system (IDS) development which using artificial immune system (AIS) with population based incremental learning (PBIL). AIS have powerful distinguished capability to extirpate antigen when the antigen intrude into human body. The PBIL is based on past learning experience to adjust new learning. Therefore we propose an intrusion detection system call PBIL-AIS which combine two approaches of PBIL and AIS to evolution computing. In AIS part we design three mechanisms such as clonal selection, negative selection and antibody level to intensify AIS performance. In experimental result, our PBIL-AIS IDS can capture high accuracy when an intrusion connection attacks.

Keywords: Artificial immune system, intrusion detection, population-based incremental learning, evolution computing.

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12 Evaluation Rabbit Serum of the Immunodominant Proteins of Mycobacterium Avium Paratuberculosis Extracts

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

Abstract:

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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11 Optimizing Spatial Trend Detection By Artificial Immune Systems

Authors: M. Derakhshanfar, B. Minaei-Bidgoli

Abstract:

Spatial trends are one of the valuable patterns in geo databases. They play an important role in data analysis and knowledge discovery from spatial data. A spatial trend is a regular change of one or more non spatial attributes when spatially moving away from a start object. Spatial trend detection is a graph search problem therefore heuristic methods can be good solution. Artificial immune system (AIS) is a special method for searching and optimizing. AIS is a novel evolutionary paradigm inspired by the biological immune system. The models based on immune system principles, such as the clonal selection theory, the immune network model or the negative selection algorithm, have been finding increasing applications in fields of science and engineering. In this paper, we develop a novel immunological algorithm based on clonal selection algorithm (CSA) for spatial trend detection. We are created neighborhood graph and neighborhood path, then select spatial trends that their affinity is high for antibody. In an evolutionary process with artificial immune algorithm, affinity of low trends is increased with mutation until stop condition is satisfied.

Keywords: Spatial Data Mining, Spatial Trend Detection, Heuristic Methods, Artificial Immune System, Clonal Selection Algorithm (CSA)

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10 Evaluation of Newly Developed Dot-ELISA Test for Identification of Naja-naja sumantrana and Calloselasma rhodostoma Venom Antigens

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

Abstract:

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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9 Effect of Combined Carbimazole and Curcuma longa Powder in Human Thyroid-Stimulating Hormone and Thyroperoxidase Antibody in Hyperthyroidism

Authors: Ahmed Abdi Hassan, Mustapha Muhammad Aliyu

Abstract:

Turmeric (Curcuma longa) belongs to the ginger family and is used for food coloring mostly in Asian countries. It has long traditional medicinal value for the treatment of inflammations with excellent antioxidant properties. The purpose of this study is to investigate the efficiency of turmeric powder in the treatment of hyperthyroidism when combined with carbimazole antithyroid drug. The trial was conducted on 20 hyperthyroid patients but only 16 of them were successfully enrolled for the study. The 16 patients were divided into two equal groups where one group was treated with the only carbimazole while the other group was treated with a combined approach of carbimazole plus turmeric for 6 months consecutively. The result obtained is promising showing an average improvement of 99% in Thyroid-stimulating hormone (TSH) and 88%Thyroid Autoantibodies (TPOAb) in patients treated with the combined approach compared to those treated with the only carbimazole with an average of 3% and 18% of TSH and TPOAb improvement respectively. However, no major difference has been observed in both T4 and T3. Therefore, turmeric powder is a promising treatment if carefully and consistently combined with carbimazole antithyroid drug at very low amounts of 1.5 to 2 grams for at least 2 to 3 times a week.

Keywords: Thyroid, curcuminoids, turmeric, thyroxine, triiodothyronine, thyroid stimulating hormone, TPOAb.

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8 Protein Profiling in Alanine Aminotransferase Induced Patient cohort using Acetaminophen

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

Abstract:

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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7 Gold-Mediated Modification of Apoferritin Surface with Targeting Antibodies

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

Abstract:

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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6 Loss of P16/INK4A Protein Expression is a Common Abnormality in Hodgkin's Lymphoma

Authors: Fawzi Irshaid, Fatiha Dilmi, Khaled Tarawneh, Raji Hadeth, Adnan Jaran, Ahad Al-Khatib

Abstract:

P16/INK4A is tumor suppressor protein that plays a critical role in cell cycle regulation. Loss of P16 protein expression has been implicated in pathogenesis of many cancers, including lymphoma. Therefore, we sought to investigate if loss of P16 protein expression is associated with lymphoma and/or any specific lymphoma subtypes (Hodgkin-s lymphoma (HL) and nonHodgkin-s lymphoma (NHL)). Fifty-five lymphoma cases consisted of 30 cases of HL and 25 cases of NHL, with an age range of 3 to 78 years, were examined for loss of P16 by immunohistochemical technique using a specific antibody reacting against P16. In total, P16 loss was seen in 33% of all lymphoma cases. P16 loss was identified in 47.7% of HL cases. In contrast, only 16% of NHL showed loss of P16. Loss of P16 was seen in 67% of HL patients with 50 years of age or older, whereas P16 loss was found in only 42% of HL patients with less than 50 years of age. P16 loss in HL is somewhat higher in male (55%) than in female (30%). In subtypes of HL, P16 loss was found exclusively in all cases of lymphocyte depletion, lymphocyte predominance and unclassified cases, whereas P16 loss was seen in 39% of mixed cellularity and 29% of nodular sclerosis cases. In low grade NHL patients, P16 loss was seen in approximately one-third of cases, whereas no or very rare of P16 loss was found in intermediate and high grade cases. P16 loss did not show any correlation with age or gender of NHL patients. In conclusion, the high rate of P16 loss seen in our study suggests that loss of P16 expression plays a critical role in the pathogenesis of lymphoma, particularly with HL.

Keywords: B-cells, immunostaining, P16 protein, Reed-Sternberg cells, tumors.

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