Search results for: ELISA

Authors: Ibrahim Aly , Rabab Zalat, Bahaa EL Deen W. El Aswad, Ismail M. Moharm , Basam M. Masoud, Tarek Diab

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The objective of the present study was to evaluate the novel nanomagnetic beads based–latex agglutination assay (NMB-LAT) as a simple test for diagnosis of S. haematobium as well as standardize the novel nanomagnetic beads based –ELISA (NMB-ELISA). According to urine examination this study included 85 S. haematobium infected patients, 30 other parasites infected patients and 25 negative control samples. The sensitivity of novel NMB-LAT was 82.4% versus 96.5% and 88.2% for NMB-ELISA and currently used sandwich ELISA respectively. The specificity of NMB-LAT was 83.6% versus 96.3% and 87.3% for NMB-ELISA and currently used sandwich ELISA respectively. In conclusion, the novel NMB-ELISA is a valuable applicable diagnostic technique for diagnosis of human schistosomiasis haematobium. The novel NMB-ELISA assay is a suitable applicable diagnostic method in field survey especially when followed by ELISA as a confirmatory test in query false negative results. Trials are required to increase the sensitivity and specificity of NMB-ELISA assay.

Keywords: Diagnosis, Latex agglutination, Nanomagnetic beads, Sandwich ELISA.

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

Abstract:

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: J. Carlos Kuri, Varun Vij, Raymond J. Turner, Orly Yadid-Pecht

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Celiac disease (CD) is an immune system disorder that is related to eating gluten. As gluten-free (GF) diet has become a concern of many people for health reasons, a gold standard had to be nominated. Enzyme-linked immunosorbent assay (ELISA) has taken the seat of this role. However, multiple limitations were discovered, and with that, the desire for an alternative method now exists. Nucleic acid based aptamers have become of great interest due to their selectivity, specificity, simplicity, and rapid-testing advantages. However, fluorescence-based aptasensors have been tagged as unstable, but lifespan details are rarely stated. In this work, the lifespan stability of a fluorescence-based aptasensor is shown over a 8-week long study displaying the accuracy of the sensor and false negatives. This study follows 22 different samples, including GF and gluten-rich (GR) and soy sauce products, off-the-shelf products, and reference material from laboratories; giving a total of 836 tests. The analysis shows an accuracy of correctly classifying GF and GR products of 96.30% and 100%, respectively, when the protocol is augmented with molecular sieves. The overall accuracy remains around 94% within the first 4 weeks and then decays to 63%.

Keywords: Aptasensor, PEG, rGO, FAM, RM, ELISA.

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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: 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: Anne-Margré C. Vink

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Background. Food-related allergies and intolerances are frequently occurring in dogs. Diagnosis and monitoring according ‘Golden Standard’ of elimination efficiency is, however, time consuming, expensive, and requires expert clinical setting. In order to facilitate rapid and robust, quantitative testing of intolerance, and determining the individual offending foods, a serological test is implicated for Alimentary Induced Diseases and manifestations. Method. As we developed Medisynx IgG Human Screening Test ELISA before and the dog’ immune system is most similar to humans, we were able to develop Medisynx IgG Dog Screening Test ELISA as well. In this randomized, double-blind, split-sample, retro perspective study 47 dogs suffering from Canine Atopic Dermatitis (CAD) and several secondary induced reactions were included to participate in serological Medisynx IgG Dog Screening Test ELISA (within < 0,02 % SD). Results were expressed as titers relative to the standard OD readings to diagnose alimentary induced diseases and monitoring efficacy of an individual eliminating diet in dogs. Split sample analysis was performed by independently sending 2 times 3 ml serum under two unique codes. Results. The veterinarian monitored these dogs to check dog’ results at least at 3, 7, 21, 49, 70 days and after period of 6 and 12 months on an individual negative diet and a positive challenge (retrospectively) at 6 months. Data of each dog were recorded in a screening form and reported that a complete recovery of all clinical manifestations was observed at or less than 70 days (between 50 and 70 days) in the majority of dogs (44 out of 47 dogs =93.6%). Conclusion. Challenge results showed a significant result of 100% in specificity as well as 100% positive predicted value. On the other hand, sensitivity was 95,7% and negative predictive value was 95,7%. In conclusion, an individual diet based on IgG ELISA in dogs provides a significant improvement of atopic dermatitis and pruritus including all other non-specific defined allergic skin reactions as erythema, itching, biting and gnawing at toes, as well as to several secondary manifestations like chronic diarrhoea, chronic constipation, otitis media, obesity, laziness or inactive behaviour, pain and muscular stiffness causing a movement disorders, excessive lacrimation, hyper behaviour, nervous behaviour and not possible to stay alone at home, anxiety, biting and aggressive behaviour and disobedience behaviour. Furthermore, we conclude that a relatively more severe systemic candidiasis, as shown by relatively higher titer (class 3 and 4 IgG reactions to Candida albicans), influence the duration of recovery from clinical manifestations in affected dogs. These findings are consistent with our preliminary human clinical studies.

Keywords: Allergy, canine atopic dermatitis (CAD), food allergens, IgG-ELISA, food-incompatibility.

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Authors: Titin Andri Wihastuti, Djanggan Sargowo

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NFκB is a transcription factor regulating many function of the vessel wall. In the normal condition , NFκB is revealed diffuse cytoplasmic expressionsuggesting that the system is inactive. The presence of activation NFκB provide a potential pathway for the rapid transcriptional of a variety of genes encoding cytokines, growth factors, adhesion molecules and procoagulatory factors. It is likely to play an important role in chronic inflamatory disease involved atherosclerosis. There are many stimuli with the potential to active NFκB, including hyperlipidemia. We used 24 mice which was divided in 6 groups. The HFD given by et libitum procedure during 2, 4, and 6 months. The parameters in this study were the amount of NFKB activation ,H2O2 as ROS and VCAM-1 as a product of NFKB activation. H2O2 colorimetryc assay performed directly using Anti Rat H2O2 ELISA Kit. The NFKB and VCAM-1 detection obtained from aorta mice, measured by ELISA kit and imunohistochemistry. There was a significant difference activation of H2O2, NFKB and VCAM-1 level at induce HFD after 2, 4 and 6 months. It suggest that HFD induce ROS formation and increase the activation of NFKB as one of atherosclerosis marker that caused by hyperlipidemia as classical atheroschlerosis risk factor.

Keywords: High Fat Diet, NFKB, H2O2, atherosclerosis

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Authors: Ali M. Mutlag, Yang Zhiqiang, Meng Jiaren, Zhang Jingyan, Li Jianxi

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The object of this study was to investigate several hormones correlated to the reproduction and inhibin A, inhibin B and NO levels in the infertile dairy cows as attempt to illustrate the physiological causes of dairy cows infertility.

40 Holstein cow (21 infertile and 19 fertile) were used at estrous phase of the cycle, Hormones FSH, LH, E2, Testosterone, were measured using ELISA method. inhibin A and B also estimated by ELISA method, Nitric oxide was measured by Greiss reagent method.

The results showed different concentrations of the hormone in which FSH illustrated significantly higher concentration in the infertile cows than fertile cows (P<0.05). LH and E2 showed significant decrease in the infertile cows than the fertile cows (P<0.05), no significant difference appeared in testosterone concentrations in the fertile cows and infertile cows (P>0.05). The both inhibins A and B showed significant P<0.05 decrease concentrations in the infertile cows also NO showed clearly significant decrease P<0.05 in the infertile cows.

In conclusion, the present study approved the poorly ovarian activities and reproduction disturbance of infertile cows in spite of trigger estrous signs, the study confirmed a positive correlation between inhibins and NO to regulate the ovarian physiology. These inhibins represent effective markers of dairy cow infertility.

Keywords: Cows, Inhibin (A, B), Infertility, Nitric oxide (NO).

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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: Ahmad Ali Shahid, Muhammad Shakil Shaukat, Kamran Shehzad Bajwa, Abdul Qayyum Rao, Tayyab Husnain

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Agriculture is the backbone of economy of Pakistan and cotton is the major agricultural export and supreme source of raw fiber for our textile industry. To combat severe problems of insect and weed, combination of three genes namely Cry1Ac, Cry2A and EPSPS genes was transferred in locally cultivated cotton variety MNH-786 with the use of Agrobacterium mediated genetic transformation. The present study focused on the molecular screening of transgenic cotton plants at T3 generation in order to confirm integration and expression of all three genes (Cry1Ac, Cry2A and EPSP synthase) into the cotton genome. Initially, glyphosate spray assay was used for screening of transgenic cotton plants containing EPSP synthase gene at T3 generation. Transgenic cotton plants which were healthy and showed no damage on leaves were selected after 07 days of spray. For molecular analysis of transgenic cotton plants in the laboratory, the genomic DNA of these transgenic cotton plants were isolated and subjected to amplification of the three genes. Thus, seventeen out of twenty (Cry1Ac gene), ten out of twenty (Cry2A gene) and all twenty (EPSP synthase gene) were produced positive amplification. On the base of PCR amplification, ten transgenic plant samples were subjected to protein expression analysis through ELISA. The results showed that eight out of ten plants were actively expressing the three transgenes. Real-time PCR was also done to quantify the mRNA expression levels of Cry1Ac and EPSP synthase gene. Finally, eight plants were confirmed for the presence and active expression of all three genes at T3 generation.

Keywords: Agriculture, Cotton, Transformation, Cry Genes, ELISA and PCR.

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Authors: Dwitya Elvira, Raveinal Masri, Rohayat Bilmahdi

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Human Immunodeficiency Virus (HIV) pandemic increased significantly worldwide. The rise in cases of HIV/AIDS was also followed by an increase in the incidence of opportunistic infection, with tuberculosis being the most opportunistic infection found in HIV/AIDS and the main cause of mortality in HIV/AIDS patients. Diagnosis of tuberculosis in HIV/AIDS patients is often difficult because of the uncommon symptom in HIV/AIDS patients compared to those without the disease. Thus, diagnostic tools are required that are more effective and efficient to diagnose tuberculosis in HIV/AIDS. CXCL-10/IP-10 is a chemokine that binds to the CXCR3 receptor found in HIV/AIDS patients with a weakened immune system. Tuberculosis infection in HIV/AIDS activates chemokine IP-10 in urine, which is used as a marker for diagnosis of infection. The aim of this study was to prove whether IP-10 urine can be a biomarker diagnosis of active lung tuberculosis in HIV-AIDS patients. Design of this study is a cross sectional study involving HIV/AIDS patients with lung tuberculosis as the subject of this study. Forty-seven HIV/AIDS patients with tuberculosis based on clinical and biochemical laboratory were asked to collect urine samples and IP-10/CXCL-10 urine being measured using ELISA method with 18 healthy human urine samples as control. Forty-seven patients diagnosed as HIV/AIDS were included as a subject of this study. HIV/AIDS were more common in male than in women with the percentage in male 85.1% vs. 14.5% of women. In this study, most diagnosed patients were aged 31-40 years old, followed by those 21-30 years, and > 40 years old, with one case diagnosed at age less than 20 years of age. From the result of the urine IP-10 using ELISA method, there was significant increase of the mean value of IP-10 urine in patients with TB-HIV/AIDS co-infection compared to the healthy control with mean 61.05 pg/mL ± 78.01 pg/mL vs. mean 17.2 pg/mL. Based on this research, there was significant increase of urine IP-10/CXCL-10 in active lung tuberculosis with HIV/AIDS compared to the healthy control. From this finding, it is necessary to conduct further research into whether urine IP-10/CXCL-10 plays a significant role in TB-HIV/AIDS co-infection, which can also be used as a biomarker in the early diagnosis of TB-HIV.

Keywords: Chemokine, IP-10 urine, HIV/AIDS, Tuberculosis.

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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: Elisa Pautasso, Matteo Castagno, Michele Osella

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In the last decade the emergence of new social needs as an effect of the economic crisis has stimulated the flourishing of business endeavours characterised by explicit social goals. Social start-ups, social enterprises or Corporate Social Responsibility operations carried out by traditional companies are quintessential examples in this regard. This paper analyses these kinds of initiatives in order to discover the main characteristics of social business models and to provide insights to social entrepreneurs for developing or improving their strategies. The research is conducted through the integration of literature review and case study analysis and, thanks to the recognition of the importance of both profits and social impacts as the key success factors for a social business model, proposes a framework for identifying indicators suitable for measuring the social impacts generated.

Keywords: Business model, case study, impacts, social business.

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Authors: T. Bovornvirakit, K. Viravaidya

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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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Authors: F. S. Abtahi, M. Koohi Hbibi, M. Khodaei Motlagh

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Bean common mosaic necrosis virus (BCMNV) is a potyvirus with a worldwide distribution. This virus causes serious economic losses in Iran in many leguminoses. During 20008, samples were collected from soybeans fields in Tehran Province. Four isolates (S1, S2 and S3) were inoculated on 15 species of Cucurbitaceae, Chenopodiaceae, Solanacae and Leguminosae. Chenopodium quinoa and C. amaranticolor. Did not developed any symptoms.all isolates caused mosaic symptoms on Phaseolus vulgaris cv. Red Kidney and P. vulgaris cv. Bountiful. The molecular weights of coat protein using SDS-PAGE and western blotting were estimated at 33 kDa. Reverse transcription polymerase chain reaction (RT-PCR) was performed using one primer pairs designed by L. XU et al. An approximately 920 bp fragment was amplified with a specific primer.

Keywords: ELISA, RT-PCR, SDS-PAGE, BCMNV.

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Authors: Kacharava Tamar, Korakhashvili Avtandil, Epitashvili Tinatin

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The article deals with the database, which was created by the authors, related to biodiversity of some families of useful plants (medicinal, aromatic, spices, dye and poisonous) existing in Georgia considering important taxonomy. Our country is also rich with endemic genera. The results of monitoring of the phytogenetic resources to reveal perspective species and situation of endemic species and resources are also discussed in this paper. To get some new medicinal and preventive treatments using plant raw material in the phytomedicine, phytocosmetics and phytoculinary, the unique phytogenetic resources should be protected because the application of useful plants is becoming irreversible. This can be observed along with intensification and sustainable use of ethnobotanical traditions and promotion of phytoproduction based on the international requirements on biodiversity (Convention on Biological Diversity - CBD). Though Georgian phytopharmacy has the centuries-old traditions, today it is becoming the main concern.

Keywords: Aromatic, medicinal, poisonous, spicy, dye plants, endemic biodiversity, endemic, ELISA, GIS.

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Authors: Y. Baba hamed, A. Medjdoub, H. Merzouk, M. Narce

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The aim of this work was to study the in vitro effects of δ-lactam 1 and its 4-chlorophenyl derivative 2, on the proliferative responses of human lymphocytes and Th1 and Th2 cytokine secretion. The possible protective role of vitamin E on intracellular stress oxidative induced by these compounds was also investigated. Peripheral blood lymphocytes were isolated using differential centrifugation on a density gradient of Histopaque. They were cultured with mitogen concanavalin A, vitamin E (10 μM) and with different concentrations of the compounds 1 and 2 (0.1 to 10 μM). Proliferation (MTT assay), IL-2, INFγ and IL-4 (Elisa kits), intracellular superoxide anion were determined. 1 and 2 were immunostimulant and increased cytokine secretion with a shift away from Th1 response to Th2. These properties were however accompanied by an increase in intracellular oxidative stress. The presence of vitamin E exhibited protective effects by reducing δ- lactam-induced superoxide anion generation in lymphocytes.

Keywords: Cytokines, δ-Lactams, In vitro Lymphocyte Proliferation, Superoxide Anion

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Authors: Novi Khila Firani, Elisa Nesdyaningtyas

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Kwashiorkor is one of nutritional problem in Indonesia, which lead to decrease immune system. This condition causes susceptibility to infectious disease, especially tuberculosis. Development of new tuberculosis vaccine will be an important strategy to eliminate tuberculosis in kwashiorkor. Previous research showed that 38-kDa Mycobacterium tuberculosis protein is one of the potent immunogen. However, the role of oral immunization with 38- kDa Mycobacterium tuberculosis protein to the number of lymphocytes in the rat model of kwashiorkor is still unknown. We used kwashiorkor rat model groups with 4% and 2% low protein diet. Oral immunization with 38-kDa Mycobacterium tuberculosis protein given with 2 booster every week. The lymphocytes number were measured by flowcytometry. There was no significant difference between the number of lymphocytes in the normal rat group and the kwashiorkor rat groups. It may reveal the role of 38-kDa Mycobacterium tuberculosis protein as a potent immunogen that can increase the lymphocytes number from kwashiorkor rat model same as normal rat.

Keywords: kwashiorkor rat, lymphocytes, 38-kDa Mycobacterium tuberculosis protein

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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: A.C. Duarte, C.I.L. Justino, K. Duarte, A.C. Freitas, R. Pereira, P. Chaves, P. Bettencourt, S. Cardoso, T.A.P. Rocha-Santos

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This paper aims at overviewing the topics of a research project (CARDIOSENSOR) on the field of health sciences (biomaterials and biomedical engineering). The project has focused on the development of a nanosensor for the assessment of the risk of cardiovascular diseases by the monitoring of C-reactive protein (CRP), which has been currently considered as the best validated inflammatory biomarker associated to cardiovascular diseases. The project involves tasks such as: 1) the development of sensor devices based on field effect transistors (FET): assembly, optimization and validation; 2) application of sensors to the detection of CRP in standard solutions and comparison with enzyme-linked immunosorbent assay (ELISA); and 3) application of sensors to real samples such as blood and saliva and evaluation of their ability to predict the risk of cardiovascular disease.

Keywords: Carbon nanotubes field effect transistors, cardiovascular diseases, C-reactive protein, sensor.

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Authors: J. Franková, V. Pivodová, F. Růžička, J. Ulrichová

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An implant elicits a biological response in the surrounding tissue which determines the acceptance and long-term function of the implant. Dental implants have become one of the main therapy methods in clinic after teeth lose. A successful implant is in contact with bone and soft tissue represent by fibroblasts. In our study we focused on the interaction between six different chemically and physically modified titanium implants (Tis-MALP, Tis-O, Tis- OA, Tis-OPAAE, Tis-OZ, Tis-OPAE) with alveolar fibroblasts as well as with five type of microorganisms (S. epidermis, S.mutans, S. gordonii, S. intermedius, C.albicans). The analysis of microorganism adhesion was determined by CFU (colony forming unite) and biofilm formation. The presence of α3β1 and vinculin expression on alveolar fibroblasts was demonstrated using phospho specific cell based ELISA (PACE). Alveolar fibroblasts have the highest expression of these proteins on Tis-OPAAE and Tis-OPAE. It corresponds with results from bacterial adhesion and biofilm formation and it was related to the lowest production of collagen I by alveolar fibroblasts on Tis-OPAAE titanium disc.

Keywords: titanium disc, alveolar fibroblasts, microorganisms, adhesion

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Authors: Elisa Orofino

Abstract:

Islamism is a very debated topic in the West but almost exclusively explored in its violent forms. Nevertheless, a number of “vocal radical Islamist” groups exist in the West and legally operate because of their non-violent nature. Vocal radicals continually inspire individuals and lead them towards specific goals and priorities, sometimes even towards violence. This paper uses the long-living group Hizb ut-Tahrir (HT) to explore the elements that make the organization appealing to segments of Muslim community in the West. This paper uses three agency variables - reflexive monitoring, the rationalization of action and the motivations for actions – to analyze HT’s appeal vis-à-vis two other Islamist groups, Ikhwan al-Muslimun and Jamaat-e-Islami (JeI), having similar goals and the same high international profile. This paper concludes that HT’s uniqueness is given by its method, detailed vision of the caliphate, consistency over time and the emphasis placed on the caliphate as the leading force of HT’s unchanged motivation for action.

Keywords: Agency, Caliphate, Radicalization, Vocal Radicals.

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Authors: Ahmad M. Khalil, Ahmad M. Alshamali, Marwan H. Gagaa

Abstract:

We investigated oxidative DNA damage caused by radio frequency radiation using 8-oxo-7, 8-dihydro-2'- deoxyguanosine (8-oxodG) generated in mice tissues after exposure to 900 MHz mobile phone radio frequency in three independent experiments. The RF was generated by a Global System for Mobile Communication (GSM) signal generator. The radio frequency field was adjusted to 25 V/m. The whole body specific absorption rate (SAR) was 1.0 W/kg. Animals were exposed to this field for 30 min daily for 30 days. 24 h post-exposure, blood serum, brain and spleen were removed and DNA was isolated. Enzyme-linked immunosorbent assay (ELISA) was used to measure 8-oxodG concentration. All animals survived the whole experimental period. The body weight of animals did not change significantly at the end of the experiment. No statistically significant differences observed in the levels of oxidative stress. Our results are not in favor of the hypothesis that 900 MHz RF induces oxidative damage.

Keywords: Mice, Mobile phone radiation, oxidative stress, 8-oxo-7, 8-dihydro-2'-deoxyguanosine

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Authors: Miguel José Oliveira, Fátima Farinha, Elisa M. J. da Silva, Rui Lança, Manuel Duarte Pinheiro, Cátia Miguel

Abstract:

The Observatory of Sustainability of the Algarve Region for Tourism (OBSERVE) will be a valuable tool to assess the sustainability of this region. The OBSERVE tool is designed to provide data and maintain an up-to-date, consistent set of indicators defined to describe the region on the environmental, sociocultural, economic and institutional domains. This ongoing two-year project has the active participation of the Algarve’s stakeholders, since they were consulted and asked to participate in the discussion for the indicators proposal. The environmental and sociocultural indicators chosen must indicate the characteristics of the region and should be in alignment with other global systems used to monitor the sustainability. This paper presents a review of sustainability indicators systems that support the first proposal for the environmental and sociocultural indicators. Others constraints are discussed, namely the existing data and the data available in digital platforms in a format suitable for automatic importation to the platform of OBSERVE. It is intended that OBSERVE will be a valuable tool to assess the sustainability of the region of Algarve.

Keywords: Sustainability, observatory, environmental indicators, sociocultural indicators, development, tourism, Algarve.

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Authors: Hala O. El-Mesallamy, Salwa M. Suwailem, Mae M. Seleem

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Visfatin and apelin are two new adipokines that recently gained a special interest in diabetes research. This study was conducted to study the interplay between these two adipokines and their correlation with other inflammatory and biochemical parameters in type 2 diabetic (T2D) postmenopausal women with CAD. Visfatin and apelin were measured by enzyme-linked immunoassay (ELISA). Visfatin was found to be significantly higher in the following groups: T2D patients without CAD, non-obese and obese T2D patients with CAD when compared to control group. Apelin was found to be significantly lower in non-obese and obese T2D patients with CAD when compared to control group. Visfatin and apelin were found to be significantly associated with each other and with other biochemical parameters. The current study provides evidence for the interplay between visfatin and apelin through the inflammatory milieu characteristic of T2D and their possible role in the pathogenesis of CAD complication of T2D. 

Keywords: Apelin, Coronary artery disease, Inflammation, Type 2 diabetes, Visfatin.

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Authors: Prajna Metta, Yanti Rusyanti, Nunung Rusminah, Bremmy Laksono

Abstract:

The decrease of neutrophil chemotaxis function may cause increased susceptibility to aggressive periodontitis (AP). Neutrophil chemotaxis is affected by formyl peptide receptor 1 (FPR1), which when activated will respond to bacterial chemotactic peptide formyl methionyl leusyl phenylalanine (FMLP). FPR1 protein value is decreased in response to a wide number of inflammatory stimuli in AP patients. This study was aimed to assess the alteration of FPR1 protein value in AP patients and if FPR1 protein value could be used as an indicator of neutrophil chemotaxis dysfunction in AP. This is a case control study with 20 AP patients and 20 control subjects. Three milliliters of peripheral blood were drawn and analyzed for FPR1 protein value with ELISA. The data were statistically analyzed with Mann-Whitney test (p>0,05). Results showed that the mean value of FPR1 protein value in AP group is 0,353 pg/mL (0,11 to 1,18 pg/mL) and the mean value of FPR1 protein value in control group is 0,296 pg/mL (0,05 to 0,88 pg/mL). P value 0,787 > 0,05 suggested that there is no significant difference of FPR1 protein value in both groups. The present study suggests that FPR1 protein value has no significance alteration in AP patients and could not be used as an indicator of neutrophil chemotaxis dysfunction.

Keywords: Aggressive periodontitis, chemotaxis dysfunction, FPR1 protein value, neutrophil.

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Authors: Donna M. Morrison, Lambert Chester, Coretta A. N. Samuels, David R. Ledoux

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A survey was conducted in the five rice-growing regions in Guyana to determine the presence of aflatoxins in multiple fractions of rice in June/October 2015 growing season. The fractions were paddy, steamed paddy, cargo rice, white rice and parboiled rice. Samples were analyzed by High Performance Liquid Chromatography. A subset of the samples was further analyzed by enzyme-linked immunosorbent assay (ELISA) for concurrence. All analyses were conducted at the University of Missouri, USA. Of the 186 samples tested, 16 had aflatoxin concentrations greater than 20 ppb the recommended limit for aflatoxins in food according to the United States Food and Drug Administration. An additional three samples had aflatoxin B₁ concentrations greater than the European Union Commission maximum levels for aflatoxin B₁ in rice at 5 µg/kg and total aflatoxins (B₁, B₂, G₁ and G₂) at 10 µg/kg. The survey indicates that there is no widespread aflatoxin problem in rice in Guyana. The incidence of aflatoxins appears to be localized.

Keywords: Aflatoxins, enzyme-linked immunosorbent assay, high-performance liquid chromatography, rice fractions.

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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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